Microbial

J Leukoc Biol, 2002 Mar, 71(3), 401 - 9
Emerging evidence that molecules expressed by mammalian tissue grafts are recognized by the innate immune system; Fox-Marsh A et al.; The innate immune system existed prior to the emergence of adaptive immunity in sharks and higher vertebrates . Homologues of many mammalian innate immune-system elements such as the toll-like receptors exist in species as distant as Drosophila . Selective pressure has led to the development of highly conserved, soluble, and cell-surface receptors that recognize functionally essential molecules shared by microbial pathogens . It is thought that molecular patterns that exquisitely distinguish pathogenic cells from mammalian cells are recognized . Therefore, it would seem unlikely that innate immune-system elements should recognize mammalian tissues . However, there is increasing evidence to suggest that this is the case and that innate immunity promotes rejection of transplanted mammalian tissues, particularly those from other species (xenografts) . Evidence for innate recognition of mammalian grafts, the nature of this recognition, and the bi-directional interactions between innate and adaptive immunity that contribute to graft rejection are discussed in this review, with the emphasis on nonvascular xenografts.

Vet Immunol Immunopathol, 2002 Feb, 85(1-2), 1 - 8
The highest priority: what microbial genomes are telling us about immunity; Palmer GH; Study of microbial genomes has provided new insight into the functions that pathogens require for survival in the animal host . Small genome bacterial pathogens, defined as those < or = 1/3 the size of Escherichia coli, include chlamydiae, rickettsiae and ehrlichiae, mycoplasmas, and spirochetes . The small genome size is believed to result from reductive evolution, a process of initial mutation with loss of function followed by progressive accumulation of mutations and eventual gene deletion . This is most notable in the 1.1 Mb genome of Rickettsia prowazekki in which 24% of the genome is non-coding, as compared to approximately 10% in the 4.4 Mb E . coli . Consequently, these pathogens are thus presumed to retain only the most important functions for survival and propagation . There is consistent evidence from small genomes that the genetic deletion is primarily related to the loss of metabolic function and especially reduction of multiple overlapping pathways and duplicated genes . Thus, these pathogens undergo progressive reduction in their genomes yet maintain the ability to infect, survive within, and cause disease in animals . In the face of this reductive process, what genes and associated functions are maintained? Strikingly, these pathogens devote a high percentage of their genomes to paralogous families of polymorphic surface molecules . This retention suggests that evasion of the immune response is the highest priority of obligate microbial pathogens and provides a strategy for identifying protective antigens for vaccine development to control disease.

Herpes, 2001 Mar, 8 Suppl 1, 2A - 5A
Immunity to herpes simplex virus: a hypothesis; Rouse BT et al.; This article discusses some possible reasons why people vary in susceptibility to primary infection by herpes simplex virus (HSV), and, in addition, why some individuals suffer lesions upon viral reactivation from latency . An underlying hypothesis is that the effectiveness of various aspects of innate immunity, conditioned by the person's microbial exposure in early life, may affect susceptibility to HSV infection, the nature of the initial adaptive immune response, and the efficacy of the memory-recall reaction following reactivation . Could domicile in a super-sanitized environment result in more troublesome problems with HSV such as is suspected to explain the rising tide of allergy?

Ann Ital Chir, 2001 Jul-Aug, 72(4), 485 - 91; discussion 491-3
Th1-like and Th2-like cytokines in patients undergoing open versus laparascopic cholecystectomy; Di Vita G et al.; The advantages of laparoscopic (LC versus, open cholecystectomy (OC) seems to be related to minimal invasive procedure and to the moderate inflammatory response . The aim of this study is to define the involvement of Th1 (IFN-gamma) and Th2 (IL-4, IL-6, IL-10, IL-13) cytokines production in vivo and in vitro in patients undergoing OC or LC . In 42 patients undergoing LC (n = 22) and OC (n = 20) Th1-like and Th2-like was evaluated before operation and at 6, 24 and 48 hours after operation for white blood cell counting and cytokines (IL-4, IL-6, IL-10, IL-13, IFN-gamma, TNF-alpha) in the sera and in the supernatants from circulating mononuclear cells stimulated with phytohemagglutinin or lipopolysaccharide . The acute phase response cytokine, IL-6, appeared significantly increased following OC than after LC . All other cytokines did not very significantly . In vitro data shows a reduction of IFN-gamma and increase in Th2-like cytokines in OC patients compared with the basal value . In LC subjects we observed an high production of IFN-gamma associated to an increase of Th2-like cytokines, like IL-10 and IL-13, even though IL-4 and IL-6 were unmodified . In contrast to OC, LC did not significantly affect immunocompetence, maintaining a moderate inflammatory response and an adequate balance between Th1 and Th2 cytokine . Furthermore, the strong activation of cells producing Th1-like cytokines in LC patients following mitogen activation indicated a consistent anti-microbial activity, that was not detectable in OC patients, that showed after activation only a Th2 response.

J Infect Dis, 2002 Feb 15, 185 Suppl 1, S66 - 72
Suppression of NF-kappaB activation by infection with Toxoplasma gondii; Shapira S et al.; The interaction of host cells with microbial products or their invasion by pathogens frequently results in activation of the NF-kappaB family of transcription factors . The studies presented here reveal that in vivo, infection with Toxoplasma gondii results in the activation of NF-kappaB . To determine whether host cells could activate NF-kappaB in response to invasion by T . gondii, Western blots, immunofluorescence, and electrophoretic mobility shift assays were used to assess the response of host cells to infection . In these studies, infection of macrophages or fibroblasts with T . gondii did not result in the activation of NF-kappaB . In addition, the ability of lipopolysaccharide to activate NF-kappaB was impaired in cultures of macrophages infected with T . gondii . Together, these data demonstrate that invasion of cells by T . gondii does not lead to the activation of NF-kappaB and suggest that the parasite may actively interfere with the pathways that lead to NF-kappaB activation.

Trends Immunol, 2002 Mar, 23(3), 130 - 5
Heat-shock proteins as activators of the innate immune system; Wallin RP et al.; Peptides bound or linked to heat-shock proteins (HSPs) of microbial or mammalian origin have been shown to elicit potent antigen-specific immunity . Some members of the HSP family, such as hsp60, hsp70, hsp90 and gp96, are able also to stimulate cells of the innate immune system directly and thus, act as 'danger'-signaling molecules . This effect is independent of HSP-associated peptides and, in many respects, resembles the effect of lipopolysaccharide (LPS) . Here, we discuss the similarities between the responses to HSPs and LPS and also, emphasize that care must be taken when working with preparations of HSPs in experimental settings and interpreting experimental data.

Immunogenetics, 2002 Feb, 53(10-11), 851 - 6 Epub 2002 Jan 22.
The promoter polymorphism of the IL-6 gene is associated with levels of antibodies to 60-kDa heat-shock proteins; Veres A et al.; Elevated levels of antibodies to 60 kDa heat shock proteins are associated with severe coronary heart disease and carotid atherosclerosis . The presence of self hsp60-reacting antibodies can only be partially explained by microbial infections and induction by bacterial hsp65 proteins, since important differences (including the epitope specificity and complement activating ability) between hsp60 and hsp65 reacting antibodies have been shown . The aim of this study was to investigate the possible effects of genetic polymorphisms of different genes of proinflammatory cytokines on anti-hsp60 autoantibody levels . One hundred and seventy-six male blood donors were recruited and antibody levels to human hsp60 and Mycobacterium bovis hsp65 were determined by ELISA . Also in these donors, polymorphisms of the promoter of the IL-6 gene at position -174, the biallelic base exchange of the IL-1 beta gene at the -511 position and the IL-1 alpha gene at position -889 were investigated by PCR . A strong association between IL-6 -174 polymorphism and anti-hsp60 antibody levels was seen; the effect on anti-hsp65 antibody was less marked . Carriers of allele C at this position had significantly lower levels of anti-hsp60 and anti-hsp65 antibodies . A lack of associations between IL-1 beta and IL-1 alpha gene polymorphisms and antibody levels was detected . This is the first study in which associations between genetic polymorphisms and autoantibody levels have been described in healthy subjects . Further studies are needed to gain insight into the detailed mechanism of how the IL-6 gene polymorphism at position -174 influences anti-hsp60 autoantibody levels.

Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2002 Feb, 93(2), 174 - 8
Patterns of microbial colonization in primary root canal infections; Siqueira JF Jr et al.; OBJECTIVE: The purpose of this study was to investigate the patterns of microbial infection of root canals in untreated teeth associated with chronic periradicular lesions by use of scanning electron microscopy . STUDY DESIGN: Fifteen extracted teeth with extensive carious lesions, radiolucent lesions of varying sizes, and attached periradicular lesions after extraction were selected for study . After fixation, lesions were removed and the teeth were split into 2 halves . The teeth were then dehydrated, sputter-coated with gold, and then examined for the patterns of microbial colonization of the root canal system by using a scanning electron microscope . RESULTS: All examined root canals were infected, and bacterial cells were seen in practically all areas of the root canal system . The pattern of colonization was not uniform between specimens and even within the same specimen . Most of the root canal walls of all specimens were heavily colonized by a root canal microbiota consisting of cocci and/or rods, often forming mixed communities . Spirilla were occasionally observed as single cells or as small clusters between other bacterial forms . Bacteria were often observed penetrating the dentinal tubules . Although a shallow penetration was the most common finding, bacterial cells could be observed reaching approximately 300 microm in some specimens . Yeastlike cells were observed in 1 specimen together with bacteria . CONCLUSIONS: The root canals of teeth associated with periradicular lesions were heavily infected by bacteria and occasionally by fungi . The pattern of colonization of the root canal microbiota often showed the characteristic of a climax community, which may require special considerations regarding its elimination and prevention of clinical problems . In addition, the observed propagation of the infection to the entire root canal system in teeth associated with periradicular lesions suggests that proper therapeutic measures may be necessary to eliminate the root canal infection completely.

J Am Acad Dermatol, 2002 Mar, 46(3), 371 - 5
Reactivity to trichophytin antigen in patients with onychomycosis: effect of terbinafine; Elewski BE et al.; BACKGROUND: Many patients with chronic dermatophytosis and onychomycosis have depressed cell-mediated immunity (CMI) to trichophytin . OBJECTIVE: The fungicidal properties of oral terbinafine provide a unique opportunity to explore whether elimination of antigen could restore CMI response in these patients . METHODS: A double-blind, placebo-controlled study evaluated the effect of terbinafine (250 mg/d for 12 weeks) on skin immunoreactivity to intradermal trichophytin antigen (TRIPA), mycologic status of the nail, and nail growth in patients with toenail onychomycosis . RESULTS: Skin reactivity, in an optimized, dose response challenge series to TRIPA was inversely related to disease chronicity . Mycologic/clinical response rates were 72%/84% for terbinafine and 0%/7% for placebo . Terbinafine increased the number of TRIPA reactors 2-fold and the mean TRIPA reaction area 4-fold; responses in placebo-treated patients were relatively unchanged . Of the 7 (of 25) patients receiving terbinafine who still had positive mycology 6 months after treatment, all were anergic to TRIPA at baseline and all but one remained so after treatment . CONCLUSION: Terbinafine treatment enhances and restores CMI to TRIPA in patients with Trichophyton rubrum onychomycosis and may thereby reduce susceptibility to reinfection . Terbinafine reversal of immunologic anergy may be an important model of microbial tolerance in chronic dermatophyte infections.

Jpn J Infect Dis, 2001 Dec, 54(6), 220 - 4
Molecular basis for innate immune recognition of microbial components; Kawasaki K et al.; Recognition of bacterial envelope constituents is one mechanism used by mammalian cells to initiate responses leading to bacterial killing, or, unfortunately, responses that also cause fatal septic shock . Many cell surface receptors by which these microbial components are recognized have been identified and characterized over the past a few years . In addition to CD14, which has been shown to be involved in the recognition of many microbial components, Toll-like receptors and MD-2 have been identified as factors playing a role in the receptor complexes of these components . Here we review the recent findings regarding the molecular basis for the recognition of microbial components.

Jpn J Infect Dis, 2001 Dec, 54(6), 209 - 19
Regulation of innate immune responses by Toll-like receptors; Takeda K et al.; Innate immune response in Drosophila is mediated by signaling through Toll receptors . In mammals, Toll-like receptors (TLRs), comprising a large family, recognize a specific pattern of microbial components . So far, the roles of TLR2, TLR4, TLR5, TLR6, and TLR9 have been revealed . The recognition of microbial components by TLRs leads to activation of innate immunity, which provokes inflammatory responses and finally the development of adaptive immunity . The inflammatory response depends on a TLR-mediated MyD88-dependent cascade . However, there seems to exist additional cascades in TLR signaling . In the case of TLR4 signaling, an MyD88-independent pathway is now being characterized . In addition to the activation of innate immune responses, TLR-mediated signaling leads to suppression of the activity of innate immune cells, represented by "lipopolysaccharide (LPS) tolerance" . Progress in elucidating the molecular mechanisms for LPS tolerance has been made through the analysis of TLR-mediated signaling pathways . Thus, the activity for innate immune responses is known to be finely regulated by TLRs.

Annu Rev Immunol, 2002, 20, 427 - 62 Epub 2001 Oct 04.
Chromatin structure and gene regulation in the immune system; Smale ST et al.; The development of the immune system and the host response to microbial infection rely on the activation and silencing of numerous, differentially expressed genes . Since the mid-1980s, a primary goal has been to identify transcription factors that regulate specific genes and specific immunological processes . More recently, there has been a growing appreciation of the role of chromatin structure in gene regulation . Before most activators of a gene access their binding sites, a transition from a condensed to a decondensed chromatin structure appears to take place . The activation of transcription is then accompanied by the remodeling of specific nucleosomes . Conversely, the acquisition of a more condensed chromatin structure is often associated with gene silencing . Chromatin structure is a particularly significant contributor to gene regulation because it is likely to be a major determinant of cell identity and cell memory . That is, the propagation of decondensed chromatin at specific loci through DNA replication and cell division helps a cell remember which genes are expressed constitutively in that cell type or are poised for expression upon exposure to a stimulus . Here we review recent progress toward understanding the role of chromatin in the immune system . The interleukin-4 gene serves as a primary model for exploring the events involved in the acquisition and heritable maintenance of a decondensed chromatin structure . Studies of the interleukin-12 p40 and interferon-beta genes are then reviewed for insight into the mechanisms by which the remodeling of specific nucleosomes in the vicinity of a promoter can contribute to rapid activation following cell stimulation . Finally, basic principles of gene silencing are discussed.

Blood, 2002 Mar 1, 99(5), 1785 - 93
Secretion of heparin-binding protein from human neutrophils is determined by its localization in azurophilic granules and secretory vesicles; Tapper H et al.; Human neutrophils have an important role in host defense against microbial infection . At different stages of an infectious process, neutrophils progressively up-regulate receptors and release various effector molecules . These are stored in several distinct types of granules with varying propensity to be secreted . Heparin-binding protein (HBP), also known as CAP37 or azurocidin, is a multifunctional, inactive serine-protease homologue . The present work shows that HBP is released from neutrophils on stimulation with secretagogues that do not trigger the secretion of azurophilic granule content . Therefore, the subcellular localization of HBP was investigated in more detail . Immunofluorescence microscopy revealed that HBP was localized close to the plasma membrane . Further analysis by fractionation of postnuclear supernatants from cavitated neutrophils showed that HBP is stored in azurophilic granules and secretory vesicles but that it is also detected to a minor extent in the plasma membrane . These findings were confirmed by immunoelectron microscopy showing that HBP colocalized with marker proteins of azurophilic granules and secretory vesicles . The presence of HBP in secretory vesicles possibly depends on the stage of cell differentiation, since the promyelocytic cell line HL-60 contains less HBP than mature neutrophils, stored exclusively in the less easily mobilized azurophilic granules . Our findings suggest that HBP can be synthesized or targeted to easily mobilized compartments at a late stage of neutrophil maturation . The ability of neutrophils to secrete HBP from secretory vesicles may be important for proinflammatory functions of this protein, such as the alteration of vascular permeability.

Aliment Pharmacol Ther, 2002 Feb, 16(2), 315 - 24
High rate of post-therapeutic resistance after failure of macrolide-nitroimidazole triple therapy to cure Helicobacter pylori infection: impact of two second-line therapies in a randomized study; Peitz U et al.; BACKGROUND: The optimal second-line treatment after failed Helicobacter pylori therapy has not been established . AIMS: To ascertain whether quadruple therapy or triple therapy with omeprazole, clarithromycin and amoxicillin is the superior re-treatment after triple therapy containing a macrolide and a nitroimidazole, and to determine the impact of microbial in vitro resistance . METHODS: Patients after failed triple therapy were randomly allocated to one of two 1-week second-line treatments: omeprazole, 40 mg, clarithromycin, 500 mg, and amoxicillin, 1 g, all b.d.; or omeprazole, 20 mg b.d., bismuth subsalicylate, 600 mg q.d.s., metronidazole, 400 mg t.d.s., and tetracycline, 500 mg q.d.s . Post-therapeutic Helicobacter pylori status was assessed by 13C-urea breath test at least 4 weeks after treatment . RESULTS: The study was terminated after including 84 patients . H . pylori cure rates differed significantly: omeprazole-clarithromycin-amoxicillin: intention-to-treat, 43%; per protocol, 50%; omeprazole-bismuth subsalicylate-metronidazole-tetracycline: intention-to-treat, 68%; per protocol, 69% . The frequencies of resistance after first-line therapy were: metronidazole, 90%; clarithromycin, 71%; both combined, 68% . For clarithromycin resistance, H . pylori cure with omeprazole-clarithromycin-amoxicillin was 30% vs . 83% for clarithromycin susceptibility . CONCLUSIONS: Omeprazole-bismuth subsalicylate-metron- idazole-tetracycline was superior to omeprazole-clarithromycin-amoxicillin, but both therapies yielded unsatisfactory results . The high rate of post-therapeutic dual resistance has a negative impact on omepraz- ole-clarithromycin-amoxicillin, and probably also on omeprazole-bismuth subsalicylate-metronidazole-tetracycline, and limits the choice for second-line treatment.

Science, 2002 Feb 22, 295(5559), 1514 - 6
Microbial activity at gigapascal pressures; Sharma A et al.; We observed physiological and metabolic activity of Shewanella oneidensis strain MR1 and Escherichia coli strain MG1655 at pressures of 68 to 1680 megapascals (MPa) in diamond anvil cells . We measured biological formate oxidation at high pressures (68 to 1060 MPa) . At pressures of 1200 to 1600 MPa, living bacteria resided in fluid inclusions in ice-VI crystals and continued to be viable upon subsequent release to ambient pressures (0.1 MPa) . Evidence of microbial viability and activity at these extreme pressures expands by an order of magnitude the range of conditions representing the habitable zone in the solar system.

Tex Dent J, 2000 Jun, 117(6), 36 - 45
The role of dentistry in palliative care of the head and neck cancer patient; Paunovich ED et al.; Although palliative care for the terminally ill is based on a multidimensional philosophy to provide whole-person comfort care while maintaining optimal function, it does not usually include dentistry in its team approach . Dentists can have a significant role in the care of these patients by providing total, active comfort care of the oral cavity . The function of the oral cavity is essential to the patient's ability to thrive . Therefore, alleviation of pain and prevention of infection in the oral cavity should be a priority in providing total, active comfort for the patient . The oral problems experienced by the hospice head and neck patient clearly affect the quality of his or her remaining life . Through routine assessments and interventions by a dentist on the palliative care team (Figure 4), comfort care for the patient may be improved by the maintenance of oral hygiene and procedures to hydrate the oral mucosa . In addition, routine dental assessments may identify dental disease and facilitate dental interventions for caries, periodontal disease, oral mucosal problems or prosthetic needs . Attention to such detail may reduce not only the microbial load of the mouth but the risk for pain and oral infection as well . This multidisciplinary approach to palliative care, including a dentist, may reduce the oral debilities that influence the patient's ability to speak, eat or swallow . Not only does maintenance of oral health have impact on the quality of life, which is already challenged by the disease, but it also aids in the ability of patients to thrive for whatever precious time is left to them.

Vox Sang, 2002 Jan, 82(1), 32 - 8
Microbial contamination of cord blood stem cells; Honohan A et al.; BACKGROUND AND OBJECTIVES: After storage, low levels of contaminating bacteria in standard blood components can reach bacteraemic levels, causing severe transfusion-associated sepsis . For cord blood (CB), the significance of low levels of contaminating bacteria and the optimal detection method is unknown and not supported by available guidelines . MATERIALS AND METHODS: Spiking experiments and testing of various subfractions of CB units were used to determine the behaviour of bacteria during centrifugation, freezing and thawing . For routine testing of CB, different volumes were compared for the detection of potential pathogens and micro-organisms of low pathogenicity . RESULTS: Centrifugation, as applied to CB fractionation, does not show concentration of bacteria in any particular fraction and supports the possibility of culture of waste fractions . Dimethylsulphoxide (DMSO) and freezing does not affect the viability of bacteria under the conditions used in this study . Owing to the low contamination level, a large sample volume of 20 ml was more sensitive than a 10-ml sample volume . Eighty five per cent of the isolated strains can be considered to be of low pathogenicity . CONCLUSION: When an optimal waste fraction sample volume of 20 ml was cultured, the contamination rate of CB was found to be approximately 13%, with low levels of < 1 colony-forming unit (CFU)/ml . Such levels of bacteria of low pathogenicity are expected to be of clinical importance only when CB is expanded in vitro for a prolonged period of time.

Huan Jing Ke Xue, 2001 Nov, 22(6), 14 - 9
{Values of carbon dioxide emission from different land-use patterns of alpine meadow}; Cao G et al.; The value order of carbon dioxide emission from different land-use of alpine meadow was significantly with season changing . It was found that the Potentilla fruticosa shrub meadow(C, 1871.40 g/m2) > Kobresia humilis meadow(A, 1769.63 g/m2) > Degraded Potentilla fruticosa shrub meadow(D, 1495.60 g/m2) > Degraded Kobresia humilis meadow(B, 1191.26 g/m2) during growth season, A(661.46 g/m2) > C(550.90 g/m2) > B(502.50 g/m2) > D (384.50 g/m2) in rest period; and A(2431.09 g/m2) > C(2422.30 g/m2) > D(1880.10 g/m2) > B(1694.06 g/m2) for whole year, separately . This deference not only decided the soil microbial activity and soil characters, but also have the closely relationship to shortage of frozen period.

Phytochemistry, 2002 Mar, 59(5), 479 - 88
Microbial transformation of cadina-4,10(15)-dien-3-one, aromadendr-1(10)-en-9-one and methyl ursolate by Mucor plumbeus ATCC 4740; Collins DO et al.; The sesquiterpenes cadina-4,10(15)-dien-3-one (1) and aromadendr-1(10)-en-9-one (squamulosone) (14) along with the triterpenoid methyl ursolate (21) were incubated with the fungus Mucor plumbeus ATCC 4740 . Substrates 1, 14 and ursolic acid (20) were isolated from the plant Hyptis verticillata in large quantities . M . plumbeus hydroxylated 1 at C-12 and C-14 . When the iron content of the medium was reduced, however, hydroxylation at these positions was also accompanied by epoxidation of the exocyclic double bond . In total nine new oxygenated cadinanes have been obtained . Sesquiterpene 14 was converted to the novel 2alpha,13-dihydroxy derivative along with four other metabolites . Methyl ursolate (21) was transformed to a new compound, methyl 3beta,7beta,21beta-trihydroxyursa-9(11),12-dien-28-oate (22) . Two other triterpenoids, 3beta,28-dihydroxyurs-12-ene (uvaol) (23) and 3beta,28-bis(dimethylcarbamoxy)urs-12-ene (24) were not transformed by the micro-organism, however.

Am J Infect Control, 2002 Feb, 30(1), 32 - 9
Surveillance of ventilator-associated pneumonia in very-low-birth-weight infants; Cordero L et al.; BACKGROUND: Surveillance of ventilator-associated pneumonia (VAP) is an essential part of quality patient care . Very-low-birth-weight (VLBW) infants, many with tracheal microbial colonization and bronchopulmonary dysplasia (BPD), comprise a difficult group in whom to make a diagnosis of pneumonia with the Centers for Disease Control and Prevention (CDC) criteria for infants younger than 1 year . OBJECTIVE: Our objective was to retrospectively compare VAP surveillance diagnoses made by the hospital infection control practitioner (ICP) with those made by a panel of experts with the same clinical and laboratory evidence and supportive radiologic data . A secondary objective was to compare radiologic diagnosis of pneumonia made by the general hospital radiologists, by the panel of experts, and by a pediatric radiologist from another hospital.Study Population: Thirty-seven VLBW infants identified as at risk for VAP by the ICP on the basis of a positive bacterial tracheal culture and the application of CDC criteria for the definition of pneumonia were studied . METHODS: Clinical and laboratory evidence and routine radiologic reports made by the general radiologist were reviewed independently by a panel of experts composed of 3 experienced neonatologists . Chest x-rays from the day before, day of, and day after the surveillance date were reviewed separately by the 3 neonatologists and also by a pediatric radiologist . RESULTS: After inter-reader reliability was found satisfactory (kappa's coefficient, 0.47-0.75; P <.05), the panel of neonatologists determined that the 37 VLBW infants represented 4 distinct clinical categories . Group 1 comprised 12 airway-colonized infants, aged 14 to 30 days, who on the surveillance date, albeit intubated, were asymptomatic, not treated with antibiotics, and survived . Group 2 comprised 11 airway-colonized infants, aged 7 to 42 days, who presented with equivocal clinical, laboratory, or radiologic signs of VAP and survived . Group 3 comprised 7 airway-colonized infants, aged 14 to 21 days, who were acutely ill (3 died) and had clinical and laboratory evidence of nosocomial bloodstream infection (BSI) but no radiologic signs of pneumonia . Group 4 comprised 7 infants, aged 14 to 28 days, who were acutely ill (4 died) and had clinical and laboratory evidence of infection and radiologic changes consistent with VAP.Radiologic Findings: General radiologists, neonatologists, and the pediatric radiologist agreed that none of the asymptomatic airway-colonized infants (Group 1) had VAP . General radiologists reported signs suggestive of pneumonia in 8 of 11 infants (Group 2), a finding not corroborated by the others . Everybody agreed on the absence of radiologic pneumonia in 6 of 7 patients with nosocomial BSI (Group 3) and on the presence of signs consistent with pneumonia in the remaining 7 infants (Group 4) . CONCLUSION: Surveillance diagnosis of VAP in VLBW infants is difficult because current CDC definitions are not specific for this population . Isolated positive tracheal culture alone does not distinguish between bacterial colonization and respiratory infection . Clinical and laboratory signs of VAP, mostly nonspecific, can be found in other conditions such as bronchopulmonary dysplasia and nosocomial BSI . Routine radiologic reports suggestive of pneumonia in airway-colonized infants without definitive clinical and laboratory evidence of infection could be misleading . To improve accuracy, surveillance diagnosis of VAP in special populations such as VLBW infants should be reformulated; meanwhile, ICPs should seek consultation with experienced clinicians for interpretation of data.

Curr Opin Biotechnol, 2002 Feb, 13(1), 65 - 7
Whole genome amplification--applications and advances; Hawkins TL et al.; The concept of whole genome amplification is something that has arisen in the past few years as the polymerase chain reaction (PCR) has been adapted to replicate regions of genomes that are of biological interest . The applications are many--forensic science, embryonic disease diagnosis, bioterrorism genome detection, "immortalization" of clinical samples, microbial diversity, and genotyping . Several recent papers suggest that whole genomes can be replicated without bias or non-random distribution of the target, these findings open up a new avenue to molecular biology.

Curr Opin Biotechnol, 2001 Dec, 12(6), 552 - 8
Microbial epoxide hydrolases for preparative biotransformations; Steinreiber A et al.; Epoxide hydrolases from microbial sources are highly versatile biocatalysts for the asymmetric hydrolysis of epoxides on a preparative scale . Besides kinetic resolution, which furnishes the corresponding vicinal diol and remaining non-hydrolysed epoxide in nonracemic form, enantioconvergent processes are possible: these are highly attractive as they lead to the formation of a single enantiomeric diol from a racemic oxirane . The data accumulated over recent years reveal a common picture of the substrate structure selectivity pattern of microbial epoxide hydrolases and indicate that substrates of various structural types can be selectively hydrolysed with enzymes from certain microbial sources.

Lett Appl Microbiol, 2002, 34(1), 46 - 50
Removal of heavy metals by an Aspergillus terreus strain immobilized in a polyurethane matrix; Dias MA et al.; AIMS: The aim was to investigate the biosorption of chromium, nickel and iron from metallurgical effluents, produced by a steel foundry, using a strain of Aspergillus terreus immobilized in polyurethane foam . METHODS AND RESULTS: A . terreus UFMG-F01 was immobilized in polyurethane foam and subjected to biosorption tests with metallurgical effluents . Maximal metal uptake values of 164.5 mg g(-1) iron, 96.5 mg g(-1) chromium and 19.6 mg g(-1) nickel were attained in a culture medium containing 100% of effluent stream supplemented with 1% of glucose, after 6 d of incubation . CONCLUSIONS: Microbial populations in metal-polluted environments include fungi that have adapted to otherwise toxic concentrations of heavy metals and have become metal resistant . In this work, a strain of A . terreus was successfully used as a metal biosorbent for the treatment of metallurgical effluents . SIGNIFICANCE AND IMPACT OF THE STUDY: A . terreus UFMG-F01 was shown to have good biosorption properties with respect to heavy metals . The low cost and simplicity of this technique make its use ideal for the treatment of effluents from steel foundries.

J Appl Microbiol, 2002, 92(1), 165 - 72
Early detection of spoilage moulds in bread using volatile production patterns and quantitative enzyme assays; Keshri G et al.; AIMS: Early detection of spoilage fungi (two Eurotium spp., a Penicillium chrysogenum species) in bread analogues over periods of 72 h at 25 degrees C and 0.95 water activity was evaluated using volatile production patterns, hydrolytic enzyme production, and changes in fungal populations . METHODS AND RESULTS: Using an electronic nose system it was possible to differentiate between uninoculated controls and samples contaminated with P . chrysogenum within 28 h . After 40-48 h it was possible to differentiate between the Eurotium spp., P . chrysogenum and the control using Principal Component Analysis (PCA) . Cluster analyses could differentiate between the control, P . chrysogenum and the Eurotium spp . after 40 h . Of seven hydrolytic enzymes examined after 48 h, the specific activities of three were significantly different from uninoculated control bread . There were also differences between the mould species in production of three enzymes . Penicillium chrysogenum populations increased fastest, from about 10(3) cfu g(-1) to 10(6)-10(7) cfu g(-1) after 72 h . For the Eurotium spp . this increase was slower . CONCLUSIONS: Overall, this study suggests, for the first time, that an electronic nose system using a surface polymer sensor array is able to detect qualitative changes in volatile production patterns for the early detection of the activity of spoilage moulds in bakery products . SIGNIFICANCE AND IMPACT OF THE STUDY: Potential exists for application of such systems for microbial quality assurance in intermediate moisture food products.

J Interferon Cytokine Res, 2002 Jan, 22(1), 87 - 93
Enhancement and diversification of IFN induction by IRF-7-mediated positive feedback; Levy DE et al.; Interferons (IFN) are potent components of the innate immune response to microbial infection . The genes for type I IFN (IFN-alpha and IFN-beta) are rapidly induced in response to viral infection through a mechanism that involves latent cellular transcription factors that are activated in response to innate recognition of viral components . IFN regulatory factor (IRF) proteins are key to this regulation, and their conversion from latent to active involves virus-induced serine phosphorylation . Differential utilization of distinct IRF proteins by different members of the type I IFN gene family produces a graded induction of gene expression, resulting in tight control of these cytokines through a positive feedback mechanism . Early response to virus causes secretion of a subset of IFN genes through the action of IRF-3 in conjunction with additional transcription factors, such as NF-kappaB and activator protein-1 (AP-1) (c-jun/ATF) . This early IFN acts in an autocrine manner to stimulate production of IRF-7, a transcription factor capable of activating the many additional members of the IFN-alpha gene family . The dependence of IRF-7 on virus-induced phosphorylation for its activity insures that IFN production is limited to virus-infected cells . Characterization of the cellular components involved in viral detection and IRF activation will further delineate this vital mechanism of innate immune response.

Immunobiology, 2001 Dec, 204(5), 659 - 66
Dendritic cell-based vaccination strategies: induction of protective immunity against leishmaniasis; Moll H et al.; The clinical symptoms caused by infections with Leishmania parasites range from self-healing cutaneous to uncontrolled visceral disease and depend not only on the parasite species but also on the type of the host's immune response . Infection of genetically susceptible mice with Leishmania major results in the development of disease-promoting T helper cells of type 2 (Th2) . On the other hand, healing of lesions is dependent on the induction of Th1 cells producing interferon-gamma (IFN-gamma) . The presence of interleukin 12 (IL-12) is known to be crucial for the differentiation of Th1 cells . Whereas IL-12 release and the T cell stimulatory functions of macrophages are down-regulated by L . major infection, dendritic cells (DC) exposed to L . major readily produce IL-12 and are highly potent antigen-presenting cells . Moreover, DC pulsed ex vivo with L . major antigen induce protection in otherwise susceptible mice against subsequent challenges with the parasites . The protection is long-lasting and correlates with a shift of the cytokine expression pattern towards a Th1 response . Thus, DC serve as immunomodulators in vivo and can be used as an effective adjuvant for vaccination against experimental leishmaniasis . Studies on the ability of DC to induce protective immunity to leishmaniasis may have important implications for the development of novel strategies for prophylactic and therapeutic immunizations against microbial pathogens.

Sci Total Environ, 2002 Feb 4, 284(1-3), 263 - 6
Effect of environmental contaminants on nasal lysozyme secretions; Noble RE; Human nasal secretions are comprised of lysozyme and albumin as their main protein components . Lysozyme, an anti-microbial substance, is produced by nasal serous cells while albumin is obtained, primarily, from increased nasal vasculature permeability . We measured lysozyme levels in nasal secretions following challenge by a variety of non-infectious environmental contaminants . The methodology given presents a simple and rapid method of collecting nasal secretions and determining their lysozyme content, a technique which can be used for a host of environmental irritants.

Holist Nurs Pract, 2002 Jan, 16(2), 80 - 8
The four S's of wound management: staples, sutures, Steri-Strips, and sticky stuff; Autio L et al.; Wound management is a daily occurrence in many practices . A short summary of current research in wound management is documented . Wound assessment guidelines are discussed . Included are evaluation of the patient's tetanus status, wound complexity and age, decisions as to the use of antibiotics, and closure methods . Follow-up care, patient education, and special considerations are also discussed . Billing information is considered . The objective of this article is that reader be able to: 1 . Identify several closure options for commonly seen wounds 2 . Identify selection criteria for use of Steri-Strips, sutures, and surgical glue in closing wounds and lacerations 3 . Identify criteria indicating the need for antibiotics and the most common causative microbial agents associated with wounds 4 . Identify over-the-counter (OTC) surgical glues and their popular uses 5 . Evaluate immunization status and recommend appropriate treatment for wound prophylaxis.

Roum Arch Microbiol Immunol, 1998 Jul-Dec, 57(3-4), 295 - 307
Proinflammatory cytokines production and PMN-elastase release from activated PMN cells in the periodontal disease; Drugarin D et al.; The aim of this study was to evaluate the local changes in the crevicular gingival fluid (CGF) determined by the inflammatory and immune response in periodontitis and gingivitis . The selected patients presented gingivitis (n = 9) and periodontitis: aggressive periodontitis (n = 21) and adult periodontitis (n = 8) . The crevicular fluid was provided from the gingival and periodontal pocket . The measurement of PMN-elastase in the CGF, using the ELISA method, showed a significant (p < 0.01) increase of the enzyme concentration in the aggressive periodontitis group (62.1 +/- 3.91 ng/ml) comparing to the gingivitis group (33.04 +/- 4.14 ng/ml) but also the increase (p < 0.05) of this enzyme in the adult periodontitis (43.6 +/- 2.16 ng/ml) comparing to the gingivitis, which indicated the evolutive aspects of the inflammatory reaction in these diseases . The increased production of PMN-E is the result of the activation of polymorphonuclear cells (PMN) as a reaction of the microbial attack . Degranulation and release of proteolytic enzymes including elastase, which present cytotoxic capacities, follow the activation of neutrophil granulocytes (PMN) . The activated granulocytes release proinflammatory cytokines IL-1, TNF-alpha which augment the inflammatory immune response . The aggressive periodontitis group showed an increased CGF level of IL-1 (780.4 +/- 104 pg/ml) comparing to the gingivitis group (275.5 +/- 78 pg/ml) (p < 0.01) . TNF-alpha also presented an increased level (p < 0.01) in the aggressive periodontitis group (16.3 +/- 2.3 pg/ml) comparing to the gingivitis group (4.1 +/- 1.2 pg/ml) as a consequence of the periodontium destruction and of the tissular necrosis in the former group . In conclusion, our study shows a significant increase of the PMN-elastase and proinflammatory cytokines level in CGF of patients with gingivitis and periodontitis . The intensity of the inflammatory response in these diseases is strongly correlated to the activation of the neutrophil granulocytes which release these biological active molecules that could be used as evolution markers of the disease.

Eur Respir J, 2002 Jan, 19(1), 158 - 71
The bidirectional capacity of bacterial antigens to modulate allergy and asthma; Renz H et al.; In recent decades, the prevalence of allergic diseases including bronchial asthma, hay fever and atopic dermatitis, has risen steadily in high-income countries . The underlying mechanisms for this phenomenon remain largely unknown . Since the natural mutation rate is low, altered environmental and lifestyle conditions are thought to play an important role . Epidemiological and clinical studies have provided indirect evidence that infections may prevent the development of atopy and atopic disease . This is referred to as the "hygiene hypothesis" . According to the hygiene hypothesis, viral and/or bacterial infections could inhibit the T-helper (Th)-2 immune response associated with atopic reactions by stimulating a Th-1 response involved in defence of bacterial infections and delayed-type hypersensitivity reactions . In particular, the prenatal period and early childhood are considered to be critical for the establishment and maintenance of a normal Th-1/Th-2 balance . On the other hand, several studies suggested that infections exacerbate established allergic diseases, e.g . bronchial asthma, airway hyperresponsiveness and atopic dermatitis . Therefore, viral and/or microbial infections and/or their products may have bidirectional effects on the development of allergy and asthma . This review will focus on recent findings related to the interaction between allergic disorders and infectious diseases, with the main emphasis on bacterial infections.

Adv Dent Res, 2000 Dec, 14, 69 - 75
Heterogeneity of high-molecular-weight human salivary mucins; Offner GD et al.; The existence of high-molecular-weight glycoproteins in saliva and salivary secretions has been recognized for nearly 30 years . These proteins, called mucins, are essential for oral health and perform many diverse functions in the oral cavity . Mucins have been intensively studied, and much has been learned about their biochemical properties and their interactions with oral micro-organisms and other salivary proteins . In the past several years, the major high-molecular-weight mucin in salivary secretions has been identified as MUC5B, one of a family of 11 human mucin gene products expressed in tissue-specific patterns in the gastrointestinal, respiratory, and reproductive tracts . MUC5B is one of four gel-forming mucins which exist as multimeric proteins with molecular weights greater than 20-40 million daltons . The heavily glycosylated mucin multimers form viscous layers which protect underlying epithelial surfaces from microbial, mechanical, and chemical assault . Another class of mucin molecules, the membrane-bound mucins, is structurally and functionally distinct from the gel-forming mucins . These proteins do not form multimers and can exist as both secreted and membrane-bound forms, with the latter anchored to epithelial cell membranes through a short membrane-spanning domain . In the present work, we show that two of the membrane-bound mucins, MUC1 and MUC4, are expressed in all major human salivary glands as well as in buccal epithelial cells . While the functions of these mucins in the oral environment are not understood, it is possible that they form a structural framework on the cell surface which not only is cytoprotective, but also may serve as a scaffold upon which MUC5B, and possibly other salivary proteins, assemble.

Adv Dent Res, 2000 Dec, 14, 40 - 7
Saliva and dental caries; Lenander-Lumikari M et al.; Caries is a unique multifactorial infectious disease . Our understanding of etiological factors, the progress of the disease, and the effectiveness of prophylactic procedures have led us to believe that we understand the disease . However, we still have too few answers to many questions: "Why can we not predict who will get the disease?" "Why do we not become immunized?" "How much saliva is enough?" or "Which salivary components are protective?" and "Which salivary components predispose for caries?" It is generally accepted, however, that saliva secretion and salivary components secreted in saliva are important for dental health . The final result, "caries to be or not to be", is a complex phenomenon involving internal defense factors, such as saliva, tooth surface morphology, general health, and nutritional and hormonal status, and a number of external factors-for example, diet, the microbial flora colonizing the teeth, oral hygiene, and fluoride availability . In this article, our aim is to focus on the effects of saliva and salivary constituents on cariogenic bacteria and the subsequent development of dental caries.

CLAO J, 2002 Jan, 28(1), 36 - 9
Health belief and health practice in contact lens wear--a dichotomy?
Fan DS, Houang ES, Lam DS, Wong EM, Seal D.
PURPOSE: To establish if the hygiene practice of contact lens wearers (CLWs) can be influenced by "health belief." METHODS: A nested case-control study based on questionnaires was conducted in Hong Kong over a 17-month period between 45 CLW patients with microbial keratitis and 135 matched asymptomatic CLW volunteer controls . RESULTS: There was no significant difference between keratitis patients and controls (volunteers) in the perceived benefits of optometrists' instructions generally . When asked specifically, however, keratitis patients scored significantly higher than controls in the perceived benefit of checking initially with the optometrist for the correct method of lens cleaning . Contact lens wearing patients with keratitis were confident that their care of lenses would prevent complications, but this probably represented over confidence in themselves . Patients also scored significantly more neutrally in self-efficacy . Surprisingly, patients were significantly less likely to perceive the cost of lens care as a barrier but did not use their disinfecting regimes properly . CONCLUSION: We have found that CLWs still regard their eye care professional as being the most important for giving them advice on hygiene . There is considerable opportunity to educate and influence CLWs in ways to prevent infection, but the advice given must be correct . Continuing education of optometrists is also required.

J Cell Biochem Suppl, 2001, Suppl 37, 106 - 9
Computational method to assign microbial genes to pathways; Pellegrini M et al.; We present techniques that mine fully sequenced microbial genomes for functional relationships between genes . We show that genes related by one of four techniques are more likely to belong to the same cellular pathways . Furthermore, we demonstrate that the pathway of an uncharacterized gene may be inferred from those of its functionally related partners . Therefore, we are now able to assign most of the genes within bacteria to cellular pathways .

J Biol Chem, 2002 Apr 26, 277(17), 15044 - 52 Epub 2002 Feb 12.
Blood group A glycosyltransferase occurring as alleles with high sequence difference is transiently induced during a Nippostrongylus brasiliensis parasite infection; Olson FJ et al.; Neutral mucin oligosaccharides from the small intestine of control rats and rats infected with the parasite Nippostrongylus brasiliensis were released and analyzed by gas chromatography-mass spectrometry . Infected animals expressed seven blood group A-like structures that were all absent in the control animals . The blood group A nature of these epitopes was confirmed by blood group A reactivity of the prepared mucins, of which Muc2 was one . Transferase assays and Northern blotting on small intestines from infected animals showed that an alpha-N-acetylgalactosaminyltransferase similar to the human blood group A glycosyltransferase had been induced . The expression was a transient event, with a maximum at day 6 of the 13-day-long infection . The rat blood group A glycosyltransferase was cloned, revealing two forms with an amino acid similarity of 95% . Both types had blood group A transferase activity and were probably allelic because none of 12 analyzed inbred strains carried both types . The second type was found in outbred rats and in one inbred strain . First generation offspring of inbred rats of each type were heterozygous, further supporting the allelic hypothesis . The transient induction and the large allelic variation could suggest that glycosyltransferases are part of a dynamic system altering mucins and other glycoconjugates as a protecting mechanism against microbial challenges.

Trends Mol Med, 2002 Jan, 8(1), 10 - 6
Inflammation and colorectal cancer: IBD-associated and sporadic cancer compared; Rhodes JM et al.; Ulcerative colitis and colonic Crohn's disease (together known as inflammatory bowel disease or IBD) are both associated with increased risk for colorectal cancer . Although it is customary to emphasize differences in the biology of IBD-associated and sporadic colon cancer, we believe these are far outweighed by the similarities . These similarities suggest that they might have similar pathogenic mechanisms . Because the normal colon is arguably in a continual state of low-grade inflammation in response to its microbial flora, it is reasonable to speculate that both IBD-associated and sporadic colon cancer might be the consequence of bacteria-induced inflammation.

Neurology, 2002 Feb 12, 58(3), 446 - 51
Release of soluble ICAM-5, a neuronal adhesion molecule, in acute encephalitis; Lindsberg PJ et al.; BACKGROUND: Intercellular adhesion molecule (ICAM)-5 (telencephalin) is an adhesion molecule in telencephalic neurons of the mammalian brain that binds to the leukocyte integrin CD11a/CD18 . The authors observed that human cerebral neurons also expressed ICAM-5 and that ICAM-5--mediated neuron--leukocyte binding in cultured hippocampal neurons . This led the authors to examine ICAM-5 expression during clinical CNS inflammation . METHODS: The authors found, by immunoblotting, a 115-kDa soluble form of ICAM-5 (sICAM-5) cleaved from the membrane-bound (130 kDa) ICAM-5, and established an ELISA assay to measure it . CSF samples of patients with acute encephalitis and MS were studied . RESULTS: sICAM-5 was increased in encephalitis (320 plus minus 107 ng/mL; n = 25), as compared with patients with MS (128 plus minus 10 ng/mL; n = 16) and control subjects without CNS disease (137 plus minus 6 ng/mL; n = 42) (p < 0.001) . The concentration of sICAM-5 correlated with the performance in the immediate recall task (p = 0.013) and with the leukocyte count in the CSF (p = 0.02), especially in cases caused by herpes simplex virus (HSV) (r = 0.94; p = 0.002) . CONCLUSIONS: sICAM-5 is cleaved from CNS into CSF during acute encephalitis, and it may mediate leukocyte--neuron interactions . sICAM-5 release from cerebral neurons may actively regulate immune responses and leukocyte adhesion during microbial neuroinvasion in humans during encephalitis.

Biosens Bioelectron, 2002 Mar, 17(3), 147 - 57
Biosensors in fish production and quality control; Venugopal V; Fishery products are important not only from a nutritional point of view, but also as an item of international trade and foreign exchange earner for a number of countries in the world . Fish and shellfish are highly perishable, and prone to vast variations in quality due to differences in species, environmental habitats, feeding habits, etc . In addition, they can also function as carriers of several microbial and other health hazards . Therefore, maintenance of quality is of utmost importance in production and trade of fishery products . Most of the current quality control techniques are time consuming and cumbersome . There is an excellent scope for the application of biosensors in the seafood industry including the rapidly expanding aquaculture operations for fast assessment of quality . This article discusses the scope of applications biosensors in the seafood industry.

Pest Manag Sci, 2002 Jan, 58(1), 3 - 9
Spatial variability in herbicide degradation in the subsurface environment of a groundwater protection zone; Wood M et al.; The aim of this study was to investigate the spatial variability in degradation and mineralization of atrazine and isoproturon in subsurface samples taken from sandy loam soils overlying gravel terraces which form part of a groundwater protection zone . Percussion drilling was used to obtain samples from 11 boreholes (maximum depth 3 m) . Unlabelled atrazine or isoproturon, and ring-14C-labelled atrazine or isoproturon were added to samples, incubated at 25 degrees C for up to 16 weeks, and analyzed for the residual herbicide or {14C}carbon dioxide . All samples showed the potential to degrade these herbicides, although the percentage degradation decreased by a factor of 2-3 from the surface soil to a depth of 3 m . This was associated with a decrease in organic matter content, but there was no change in the potential to mineralize acetate, indicating that specific changes in the catabolic ability of the microbial population occurred with depth . The capacity of samples to mineralize atrazine and isoproturon to carbon dioxide decreased markedly with depth, with no mineralization potential observed at a depth of 80 cm.

J Environ Qual, 2002 Jan-Feb, 31(1), 241 - 7
Degradation of 14C-atrazine bound residues in brown soil and rendzina fractions; Munier-Lamy C et al.; The remobilization and the fate of 14C-ring labeled atrazine (6-chloro-N2-ethyl-N4-isopropyl-1,3,5-triazine-2,4-diamine) bound residues was examined in relation with the turnover of natural soil organic matter . Soil fractions of a brown soil and a rendzina were incubated under controled laboratory conditions . The mineralization of natural organic matter and atrazine-bound residues was respectively estimated by the amounts of CO2 and 14CO2 evolved during the incubation . The remobilization and distribution of 14C residues among the soil organic fractions were achieved after physical-chemical extractions of the samples . Comparisons of samples in abiotic and biotic conditions allowed us to assess the influence of microbial activity on the fate of atrazine-bound residues . The mineralization curves showed that natural organic matter and atrazine-bound residues had similar decomposition patterns . After 100 d of incubation, 0.8 to 3.6% of total organic C was evolved as CO2, while only 0.1% of the initial radioactivity was mineralized as CO2, and 7 to 15% was becoming extractable with water and methanol . Few differences were observed in the distribution of residues within organic compounds for both fractions of the rendzina, except a decrease of the 14C radioactivity of the 50- to 5000-microm fraction and a slight increase of that of humin . For the 0- to 5000-microm brown soil fraction, increased radioactivity in humin at the expense of humic (HA) and fulvic (FA) acids was detected after incubation, while for the 0- to 50-microm fraction more radioactivity was recovered with FA.

Ned Tijdschr Tandheelkd, 1995 Jan, 102(1), 18 - 20
{Caries and heredity}; Konig KG; Although host genes play a certain role, the development of carious lesions is determined predominantly by local influences in the oral cavity, especially by microbial plaque and dietary carbohydrates as substrate for acidogenic bacteria . How strong or weak the influence of the hereditary component on caries really is, has been the subject of discussions for the last decades . The geneticist Professor Sofear is optimistic about prospects to decrease the caries risk by manipulation of genes . However, more detailed analysis of known facts shows that the majority of genetic differences is caused by indirectly operating mechanisms . Furthermore, these genetic factors can be overruled by local protective and compensating influences.

J Clin Lab Anal, 2002, 16(1), 47 - 51
Multiplex polymerase chain reaction: a practical approach; Markoulatos P et al.; Considerable time and effort can be saved by simultaneously amplifying multiple sequences in a single reaction, a process referred to as multiplex polymerase chain reaction (PCR) . Multiplex PCR requires that primers lead to amplification of unique regions of DNA, both in individual pairs and in combinations of many primers, under a single set of reaction conditions . In addition, methods must be available for the analysis of each individual amplification product from the mixture of all the products . Multiplex PCR is becoming a rapid and convenient screening assay in both the clinical and the research laboratory . The development of an efficient multiplex PCR usually requires strategic planning and multiple attempts to optimize reaction conditions . For a successful multiplex PCR assay, the relative concentration of the primers, concentration of the PCR buffer, balance between the magnesium chloride and deoxynucleotide concentrations, cycling temperatures, and amount of template DNA and Taq DNA polymerase are important . An optimal combination of annealing temperature and buffer concentration is essential in multiplex PCR to obtain highly specific amplification products . Magnesium chloride concentration needs only to be proportional to the amount of dNTP, while adjusting primer concentration for each target sequence is also essential . The list of various factors that can influence the reaction is by no means complete . Optimization of the parameters discussed in the present review should provide a practical approach toward resolving the common problems encountered in multiplex PCR (such as spurious amplification products, uneven or no amplification of some target sequences, and difficulties in reproducing some results) . Thorough evaluation and validation of new multiplex PCR procedures is essential . The sensitivity and specificity must be thoroughly evaluated using standardized purified nucleic acids . Where available, full use should be made of external and internal quality controls, which must be rigorously applied . As the number of microbial agents detectable by PCR increases, it will become highly desirable for practical purposes to achieve simultaneous detection of multiple agents that cause similar or identical clinical syndromes and/or share similar epidemiological features .

Curr Opin Microbiol, 2002 Feb, 5(1), 102 - 10
How Drosophila combats microbial infection: a model to study innate immunity and host-pathogen interactions; Tzou P et al.; During the past year, dramatic progress has been achieved in our understanding of Drosophila immune reactions . The completion of the Drosophila genome sequencing project, microarray analysis and the use of genetic screens have led to the identification of several new genes required to combat microbial infection, filling in some important gaps in the understanding of innate immunity . At the same time, this insect was used as a model for the study of host-pathogen interactions . The recent major advances on the mechanisms by which this insect defends itself against intrusion of pathogens are discussed in this review.

Curr Opin Microbiol, 2002 Feb, 5(1), 70 - 5
Toll-like receptors: mammalian "taste receptors" for a smorgasbord of microbial invaders; Sieling PA et al.; In Drosophila, the Toll family of proteins is responsible for the recognition of bacteria and fungi . In mammals, Toll-like receptors (TLRs) are able to recognize and respond to microbial pathogens . Recent findings have defined the relationship between many TLRs and their microbial ligands, as well as the effect of TLR ligation on host defense . These findings have also provided a framework for determining how TLRs may by used to therapeutically modulate immune responses to infection.

Curr Opin Microbiol, 2002 Feb, 5(1), 27 - 32
Genomic analyses of microbial virulence; Sassetti C et al.; Genomic sequencing of bacterial pathogens is providing an increasing wealth of new data . It is, however, still unclear how this information can be used to develop new experimental approaches . Here, we describe recent efforts to complement existing bacterial genetics with genomic methods for the study of pathogenesis.

Water Environ Res, 2001 Nov-Dec, 73(6), 704 - 10
Comparison of fatty acid composition and kinetics of phosphorus-accumulating organisms and glycogen-accumulating organisms; Wang JC et al.; It was demonstrated that glycogen-accumulating organisms (GAOs) were able to compete with phosphorus-accumulating organisms (PAOs) for acetate in a biological phosphorus removal (BPR) process, leading to a loss of BPR capability . Cellular fatty acid composition, which serves as a fingerprint for microbial identification, was used to determine microbial population change and to investigate the competition mechanisms of PAOs and GAOs . Analysis of cellular fatty acid composition indicated that PAOs grown with acetate and glucose were different species and that GAOs and PAOs grown with the same substrate were also different species . Glycogen-accumulating organisms seemed to coexist with PAOs even in a well-developed BPR process . The GAOs were able to accumulate more poly-beta-hydroxybutyrate (PHB) and glycogen than PAOs during the anaerobic stage of the BPR process . The GAOs synthesized more in-cell glycogen than PAOs . The growth rate for PAOs was always greater than that for GAOs at various acetate or glucose concentrations, while GAOs had higher acetate uptake and PHB synthesis rates than PAOs . Therefore, GAOs are thought to compete with PAOs only at long solids retention times (> or = 20 days).

Nature, 2002 Feb 7, 415(6872), 630 - 3
Unsuspected diversity among marine aerobic anoxygenic phototrophs; Beja O et al.; Aerobic, anoxygenic, phototrophic bacteria containing bacteriochlorophyll a (Bchla) require oxygen for both growth and Bchla synthesis . Recent reports suggest that these bacteria are widely distributed in marine plankton, and that they may account for up to 5% of surface ocean photosynthetic electron transport and 11% of the total microbial community . Known planktonic anoxygenic phototrophs belong to only a few restricted groups within the Proteobacteria alpha-subclass . Here we report genomic analyses of the photosynthetic gene content and operon organization in naturally occurring marine bacteria . These photosynthetic gene clusters included some that most closely resembled those of Proteobacteria from the beta-subclass, which have never before been observed in marine environments . Furthermore, these photosynthetic genes were broadly distributed in marine plankton, and actively expressed in neritic bacterioplankton assemblages, indicating that the newly identified phototrophs were photosynthetically competent . Our data demonstrate that planktonic bacterial assemblages are not simply composed of one uniform, widespread class of anoxygenic phototrophs, as previously proposed; rather, these assemblages contain multiple, distantly related, photosynthetically active bacterial groups, including some unrelated to known and cultivated types.

Microbiology, 2002 Feb, 148(Pt 2), 443 - 51
Phage-display antibody detection of Chlamydia trachomatis-associated antigens; Lindquist EA et al.; A phage-displayed human single-chain Fv antibody library (6.7x10(9) members) was used to select probes specific to components associated with the surface of Chlamydia trachomatis elementary bodies (EBs) . Each of 15 antibodies was characterized by ELISA, dot-blot, immunoblot and immunocytochemistry, resulting in the identification of several new chlamydial components associated with the surface of EBs . In addition, six antibodies were specific for host-cell components associated with the surface of EBs . While phage display has been used effectively to produce specific antibodies for purified components, these data show that this technology is suitable for selection of specific probes from complex antigens such as the surface of a microbial pathogen.

Appl Microbiol Biotechnol, 2002 Jan, 58(1), 23 - 9
Biosynthesis, biotechnological production and applications of 5-aminolevulinic acid; Sasaki K et al.; Microbial production of 5-aminolevulinic acid (ALA) by photosynthetic bacteria compared to other bacteria and algae is reviewed . During aerobic-microaerobic cultivation of Rhodobacter sphaeroides mutant strain CR520, control of the redox potential was effective for producing large amounts of extracellular ALA . ALA has been practically applied in agriculture as an herbicide, an insecticide and a growth-promoting factor for plants . New agricultural applications including salt tolerance and cold temperature tolerance of plants are also described . Finally, recent medical applications for cancer treatment, tumor diagnosis and other clinical uses are discussed.

Indian J Exp Biol, 2001 Sep, 39(9), 906 - 10
Rapid extraction of DNA from diverse soils by guanidine thiocyanate method; Agarwal A et al.; Molecular methods are being frequently used for the study of soil microbial communities as majority of naturally occurring microbial populations are non-culturable . In the present study, we describe a protocol of DNA extraction from diverse soils using a combination of heat, enzyme (lysozyme) and guanidine thiocyanate . The efficacy of the procedure was evaluated in terms of yield, purity and duration of extraction . The protocol was effective for neutral, acidic as well as alkaline soils (pH range 4.5-8.5) . The extracted soil DNA was observed with negligible shearing on agarose gel and the time taken for restriction digestion was very less . Further, the DNA extracted was almost completely devoid of contaminants and pure enough which could be used for PCR amplification and Southern hybridization.

Biochem J, 2002 Feb 15, 362(Pt 1), 231 - 7
Allotopic antagonism of the non-peptide atrial natriuretic peptide (ANP) antagonist HS-142-1 on natriuretic peptide receptor NPR-A; Poirier H et al.; The microbial polysaccharide HS-142-1 has been documented as an antagonist of natriuretic peptides . It inhibits activation and peptide binding to both guanylate receptors natriuretic peptide receptor (NPR)-A and NPR-B, but has no effect on the non-cyclase receptor NPR-C . At first sight the effect of HS-142-1 on peptide binding appears to be surmountable, suggesting that it might be competitive despite its chemically divergent nature . We explored its mode of action on wild-type NPR-A (WT), on a disulphide-bridged constitutively active mutant (C423S) and on truncated mutants lacking either their cytoplasmic domain (DeltaKC) or both the cytoplasmic and the transmembrane domains (ECD) . On the WT, HS-142-1 inhibited atrial natriuretic peptide (ANP) binding with a pK value of 6.51 +/- 0.07 (K(d)=0.31 microM) . It displayed a similar effect on the C423S mutant (pK=6.31 +/- 0.11), indicating that its action might not be due to interference with receptor dimerization . HS-142-1 also inhibited ANP binding to DeltaKC with a pK of 7.05 +/- 0.05 (K(d)=0.089 microM), but it was inactive on ANP binding to ECD at a concentration of 10(-4) M, suggesting that the antagonism was not competitive at the peptide-binding site located on the ECD and that the transmembrane domain might be required . HS-142-1 also enhanced dissociation of NPR-A-bound (125)I-ANP in the presence of excess unlabelled ANP, implying an allotopic (allosteric) mode of action for the antagonist.

Analyst, 2002 Jan, 127(1), 98 - 103
Monoclonal-based enzyme-linked immunosorbent assay and immunochromatographic assay for enrofloxacin in biological matrices; Watanabe H et al.; Enrofloxacin has been increasingly used in veterinary medicine to treat microbial infections . A simple and reliable analytical method for this drug is required . The current determination by high performance liquid chromatography (HPLC) is sensitive but labor-intensive . This paper reports an enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody (MAb) and the development of a rapid test kit based on immunochromatography . The detection limits using the ELISA were 10 ppb for chicken liver and muscle, and 1 ppb for cattle milk, respectively . The mean recovery values were 77.3-96.0% for chicken liver, 72.4-92.0% for chicken muscle and 84.0-99.0% for cattle milk . The detection limits using the kit were ca . 100 ppb for chicken muscle and ca . 10 ppb for cattle milk, respectively . All ELISA results for assay of chicken liver, chicken muscle and cattle milk were confirmed using HPLC which is used as the routine assay . The HPLC (x) and ELISA (y) results showed close correlation for chicken liver (y = 8.7 + 0.85x, r2 = 0.99, n = 25), chicken muscle (y = -3.9 + 0.94x, r2 = 0.98, n = 25) and cattle milk (y = 18.4 + 0.92x, r2 = 0.99, n = 25).

Water Res, 2002 Feb, 36(3), 713 - 21
Impact on the water column biogeochemistry of a Mediterranean mussel and fish farm; La RT et al.; We investigated and compared the impact of organic loads due to the biodeposition of mussel and fish farms on the water column of a coastal area of the Tyrrhenian Sea (Western Mediterranean) . Physico-chemical data (including oxygen, nutrients . DOC and particulate organic matter), microbial variables (picoplankton and picophytoplankton density and biomass) and phytoplankton biomass (as chlorophyll-a) were determined on a monthly basis from March 1997 to February 1998 . The results of this study indicate that both fish farm and mussel culture did not alter significantly dissolved inorganic phosphorus and chlorophyll-a values, while inorganic nitrogen concentrations were higher in mussel farm area . However, waters overlying the fish farm presented significantly higher DOC concentrations . In contrast, no significant differences were observed comparing particulate matter concentrations . The increased DOC concentrations determined a response of the heterotrophic fraction of picoplankton, while picophytoplankton, likewise phytoplankton . did not display differences among fish or mussel farms and control site . From the analysis of the different microbial components, it is possible to conclude that the impact of fish farms is evident only for the heterotrophic components . The comparative analysis of the mussel biodeposition and fish-farm impact revealed that mussel farms induced a considerably lower disturbance, apparently limited to an increased density and biomass of microbial assemblages beneath the mussel cultures.

J Antibiot (Tokyo), 2001 Nov, 54(11), 962 - 6
Selective production of staplabin and SMTPs in cultures of Stachybotrys microspora fed with precursor amines; Hu W et al.; Staplabin and SMTPs, a family of triprenyl phenol metabolites of Stachybotrys microspora, enhance fibrinolysis by modulating plasminogen conformation to increase its susceptibility to activation by plasminogen activators . We found that the production of these metabolites were markedly elevated by feeding the microbial culture with an amino acid or an amino alcohol that is a partial molecular constituent of the compound . Thus, the addition of 5-aminovaleric acid, 2-aminoethanol, Ser, Phe, Leu, Trp, Orn and Lys at 100 mg/ml resulted in 7- to 45-fold increases in the production of staplabin, SMTP-1, -3, -4, -5, -6, -7 and -8, respectively . Although the feeding at day 0 to 3 of culture supported the selective production, the supplementation after 5 days had little or no effect . When non-constituent amino acids were supplemented to cultures, production of hitherto uncharacterized congeners was observed.

Hepatology, 2002 Feb, 35(2), 403 - 8
Fine specificity of autoantibodies to soluble liver antigen and liver/pancreas; Herkel J et al.; Autoantibodies to soluble liver antigen and liver pancreas (SLA/LP) have been described as specific markers for Autoimmune Hepatitis (AIH), occurring in about 20% of patients with AIH . The high degree of specificity for SLA/LP in autoimmune liver disease suggests a possible role in its pathogenesis . This study aims to map the exact epitope(s) recognized by SLA/LP autoantibodies and to assess the role of molecular mimicry between microbial antigens and self-epitopes . Using SLA/LP-reactive sera of 18 individual AIH patients and a pool of 15 patient sera, we found the dominant immune reactivity directed to peptide p395-414 and a less prominent immune response to 2 other epitopes adjacent to the dominant epitope . Immunodominance of peptide p395-414 was confirmed by absorption experiments . The SLA/LP autoantibodies of all tested AIH patients were mainly of the IgG1 type, suggesting that SLA/LP autoantibodies may arise by a common and specific underlying immune stimulus . Based on sequence homologies of the SLA/LP antigenic region with viral proteins, it was hypothesized that molecular mimicry may drive autoimmunity to SLA/LP . However, the homologous virus-derived peptides were not recognized by SLA/LP autoantibodies . Similarly, the only known procaryotic homologue, MJ0610 of Methanococcus jannaschii, was only weakly recognized by SLA/LP-positive sera . Thus, no evidence could be found for molecular mimicry being the causative mechanism for the development of SLA/LP autoantibodies . In conclusion, the exquisite epitope specificity and IgG subtype are evidence for the maturity of the SLA/LP autoantibody response; a specific autoantigen-driven process underlying the immunopathogenesis is likely.

Trends Microbiol, 2001 Nov, 9(11), 535 - 40
Where are the pseudogenes in bacterial genomes?
Lawrence JG, Hendrix RW, Casjens S.
Most bacterial genomes have very few pseudogenes; notable exceptions include the genomes of the intracellular parasites Rickettsia prowazekii and Mycobacterium leprae . As DNA can be introduced into microbial genomes in many ways, the compact nature of these genomes suggests that the rate of DNA influx is balanced by the rate of DNA deletion . We propose that the influx of dangerous genetic elements such as transposons and bacteriophages selects for the maintenance of relatively high deletion rates in most bacteria; the sheltered lifestyle of intracellular parasites removes this threat, leading to reduced deletion rates and larger pseudogene loads.

Chem Pharm Bull (Tokyo), 2002 Jan, 50(1), 59 - 65
Microbial enantioselective ester hydrolysis for the preparation of optically active 4,1-benzoxazepine-3-acetic acid derivatives as squalene synthase inhibitors; Tarui N et al.; Microbial enantioselective ester hydrolysis for the preparation of optically active (3R,5S)-(-)-5-phenyl-4,1-benzoxazepine-3-acetic acid derivatives as potent squalene synthase inhibitors was investigated . Pseudomonas diminuta and Pseudomonas taetrolens hydrolyzed the racemic ethyl ester of the 5-(2-chlorophenyl) analogue to yield the (-)-carboxylic acid with excellent enantiomeric excess (>99% ee) . We found that the (-)-enantiomer was an active inhibitor . Bulkiness of the ester moiety did not affect the enantioselectivity but did affect reactivity . The racemic ethyl ester of the 5-(2-methoxyphenyl) analogue, 5-(2,3-dimethoxyphenyl) analogue and 5-(2,4-dimethoxyphenyl) analogue were also hydrolyzed with Pseudomonas taetrolens to afford enantiomerically pure (-)-carboxylic acids in large scale . As another route to (3R,5S)-(-)-7-chloro-5-(2,3-dimethoxyphenyl)-1-neopentyl-2-oxo-1,2,3,5-tetrahydro-4,1-benzoxazepine-3-acetic acid {(-)-1c}, the earlier intermediate (-)-2-amino-5-chloro-alpha-(2,3-dimethoxyphenyl)benzyl alcohol {(-)-12} was successfully obtained by asymmetric hydrolysis of (+/-)-5-chloro-alpha-(2,3-dimethoxyphenyl)-2-pivaloylaminobenzyl acetate with Pseudomonas sp . S-13 with >99% ee in kilogram scale followed by alkaline treatment . The product (-)-12 was converted to (-)-1c without racemization.

J Dent Res, 2002 Jan, 81(1), 53 - 7
The microbiota of young children from tooth and tongue samples; Tanner AC et al.; This study determined the frequency with which 38 microbial species were detected in 171 randomly selected children from 6 to 36 months of age . Children were sampled and dental caries measured . Oral samples were assayed by means of a checkerboard DNA probe assay . The detection frequencies from tongue samples in children under 18 mos were: S . mutans 70%, S . sobrinus 72%, P . gingivalis 23%, B . forsythus 11%, and A . actinomycetemcomitans 30%, with similar detection frequencies in children over 18 mos . Thus, S . mutans and the periodontal pathogens, P . gingivalis and B . forsythus, were detected even in the youngest subjects . Species associated with caries included S . mutans (children ages 18-36 mos) and A . israelii (children ages < 18 mos), the latter species possibly reflecting increased plaque in children with caries . Species detection from tooth and tongue samples was highly associated, with most species detected more frequently from tongue than from tooth samples in children under 18 mos, suggesting that the tongue was a potential microbial reservoir.

Org Lett, 2002 Jan 24, 4(2), 197 - 9
Militarinone A, a neurotrophic pyridone alkaloid from Paecilomyces militaris; Schmidt K et al.; {structure: see text} A new pyridone alkaloid, militarinone A (1), was isolated by bioassay-guided fractionation from the mycelium of the entomogenous fungus Paecilomyces militaris . Its structure was established by extensive spectroscopic analysis . The compound features an unprecedented side chain and a 1,4-substituted cyclohexyl moiety not previously encountered in microbial metabolites . Militarinone A had a pronounced neurotrophic effect in PC-12 cells at 10 microM concentrations.

J Immunol, 2002 Feb 15, 168(4), 1911 - 8
Eotaxin-2 generation is differentially regulated by lipopolysaccharide and IL-4 in monocytes and macrophages; Watanabe K et al.; The eotaxins are a family of CC chemokines that coordinate the recruitment of inflammatory cells, in particular eosinophils, to sites of allergic inflammation . The cDNA for eotaxin-2 (CC chemokine ligand 24) was originally isolated from an activated monocyte library . In this study, we show for the first time that peripheral blood monocytes generate bioactive eotaxin-2 protein constitutively . Eotaxin-2 production was significantly up-regulated when monocytes were stimulated with the proinflammatory cytokine IL-1beta and the microbial stimuli, LPS and zymosan . In contrast, the Th2 cytokines, IL-4 and IL-13, and the proinflammatory cytokine, TNF-alpha, acting alone or in combination, did not enhance the generation of eotaxin-2 by monocytes . Indeed, IL-4 suppressed the generation of eotaxin-2 by LPS-stimulated monocytes . Although other chemokines, including macrophage-inflammatory protein-1alpha, monocyte chemoattractant protein-1, macrophage-derived chemokine, and IL-8 were generated by monocytes, eotaxin-1 (CC chemokine ligand 11) could not be detected in the supernatants of monocytes cultured in the presence or absence of any of the stimuli used in the above experiments . Furthermore, human dermal fibroblasts that produce eotaxin-1 did not generate eotaxin-2 under basal conditions or when stimulated with specific factors, including IL-4, IL-13, TNF-alpha, and LPS . When monocytes were differentiated into macrophages, their constitutive generation of eotaxin-2 was suppressed . Moreover, IL-4, but not LPS, up-regulated the production of eotaxin-2 by macrophages . Taken as a whole, these results support a role for macrophage-derived eotaxin-2 in adaptive immunity, with a Th2 bias . In contrast, a role for monocyte-derived eotaxin-2 is implicated in innate immunity.

Appl Environ Microbiol, 2002 Feb, 68(2), 963 - 7
Microbial growth inside insulated external walls as an indoor air biocontamination source; Pessi AM et al.; The association between moisture-related microbial growth (mesophilic fungi and bacteria) within insulated exterior walls and microbial concentrations in the indoor air was studied . The studied apartment buildings with precast concrete external walls were situated in a subarctic zone . Actinomycetes in the insulation layer were found to have increased concentrations in the indoor air . The moisture content of the indoor air significantly affected all measurable airborne concentrations.

Appl Environ Microbiol, 2002 Feb, 68(2), 838 - 45
Molecular relationship between two groups of the genus Leptospirillum and the finding that Leptospirillum ferriphilum sp . nov . dominates South African commercial biooxidation tanks that operate at 40 degrees C; Coram NJ et al.; Iron-oxidizing bacteria belonging to the genus Leptospirillum are of great importance in continuous-flow commercial biooxidation reactors, used for extracting metals from minerals, that operate at 40 degrees C or less . They also form part of the microbial community responsible for the generation of acid mine drainage . More than 16 isolates of leptospirilla were included in this study, and they were clearly divisible into two major groups . Group I leptospirilla had G+C moles percent ratios within the range 49 to 52% and had three copies of rrn genes, and based on 16S rRNA sequence data, these isolates clustered together with the Leptospirillum ferrooxidans type strain (DSM2705 or L15) . Group II leptospirilla had G+C moles percent ratios of 55 to 58% and had two copies of rrn genes, and based on 16S rRNA sequence data, they form a separate cluster . Genome DNA-DNA hybridization experiments indicated that three similarity subgroups were present among the leptospirilla tested, with two DNA-DNA hybridization similarity subgroups found within group I . The two groups could also be distinguished based on the sizes of their 16S-23S rRNA gene spacer regions . We propose that the group II leptospirilla should be recognized as a separate species with the name Leptospirillum ferriphilum sp . nov . Members of the two species can be rapidly distinguished from each other by amplification of their 16S rRNA genes and by carrying out restriction enzyme digests of the products . Several, but not all, isolates of the group II leptospirilla, but none from group I (L . ferrooxidans), were capable of growth at 45 degrees C . All the leptospirilla isolated from commercial biooxidation tanks in South Africa were from group II.

Appl Environ Microbiol, 2002 Feb, 68(2), 661 - 7
Comparison of fluorescently labeled oligonucleotide and polynucleotide probes for the detection of pelagic marine bacteria and archaea; Pernthaler A et al.; We compared the detection of bacteria and archaea in the coastal North Sea and at Monterey Bay, Calif., after fluorescence in situ hybridization (FISH) either with rRNA-targeted oligonucleotide probes monolabeled with the cyanin dye Cy3 (oligoFISH) or with fluorescein-labeled polyribonucleotide probes (polyFISH) . During an annual cycle in German Bight surface waters, the percentages of bacteria visualized by polyFISH (annual mean, 77% of total counts) were significantly higher than those detected by oligoFISH (53%) . The fraction of total bacteria visualized by oligoFISH declined during winter, whereas cell numbers determined by polyFISH remained constant throughout the year . Depth profiles from Monterey Bay showed large differences in the fraction of bacterial cells visualized by polyFISH and oligoFISH in the deeper water layers irrespective of the season . Image-analyzed microscopy indicated that the superior detection of cells by polyFISH with fluorescein-labeled probes in bacterioplankton samples was less a consequence of higher absolute fluorescence intensities but was rather related to quasi-linear bleaching dynamics and to a higher signal-to-background ratio . The relative abundances of archaea in North Sea and Monterey Bay spring samples as determined by oligoFISH were on average higher than those determined by polyFISH . However, simultaneous hybridizations with oligonucleotide probes for bacteria and archaea suggested that the oligoFISH probe ARCH915 unspecifically stained a population of bacteria . Using either FISH technique, blooms of archaea were observed in North Sea surface waters during the spring and summer months . Marine group II archaea (Euryarchaeota) reached >30% of total picoplankton abundances, as determined by polyFISH . We suggest that studies of pelagic microbial community structure using oligoFISH with monolabeled probes should focus on environments that yield detections > or =70% of total cell counts, e.g., coastal surface waters during spring and summer.

Environ Pollut, 2002, 116(3), 405 - 11
Tillage, crop rotation, and organic amendment effect on changes in soil organic matter; Rickman R et al.; Carbon sequestration in agricultural soils is controlled by the balance of added organic residues and microbial oxidation of both residues and native organic matter (OM) as moderated by management and tillage . The PC-based model CQESTR predicts decomposition of residues, organic amendments and soil OM, based on cropping practices . CQESTR uses RUSLE (Revised Universal Soil Loss Equation) crop rotation and management practice, crop production, and operation databases . These data are supplemented with residue nitrogen and soil OM, bulk density, and layer thickness . CQESTR was calibrated with soil carbon data from 70-year-long experiments at the Research Center at Pendleton, OR . The calibrated model provides estimates with a 95% confidence interval of 0.33% OM . Validation at 11 independent sites resulted in a matching of observed with calculated OM with a 95% confidence interval of 0.55% OM . A 12th site, with a history of severe erosion, provided a poor match.

FEMS Immunol Med Microbiol, 2002 Jan 14, 32(2), 149 - 58
Differential microbial clearance and immunoresponse of Balb/c (Nramp1 susceptible) and DBA2 (Nramp1 resistant) mice intracerebrally infected with Mycobacterium bovis BCG (BCG); Mazzolla R et al.; In mice, the gene encoding Nramp1 (natural resistance-associated protein 1) exists in two allelic forms, differing for a point mutation . According to Nramp1 genotype, extensive literature documents a clear-cut distinction of inbred strains in two non-overlapping groups that phenotypically express resistance (Nramp1r) and susceptibility (Nramp1s) to systemic infections . Here, we provide evidence that Nramp1r (DBA/2) and Nramp1s (Balb/c) mice differently handle intracerebral infection with Mycobacterium bovis BCG . Distinct trends of microbial clearance from the brain and also different patterns of local immune responses occur, thus arguing on the involvement of Nramp1 gene product on the accomplishment of cerebral anti-mycobacterial defenses.

Fertil Steril, 2002 Feb, 77(2), 260 - 9
Are zinc levels in seminal plasma associated with seminal leukocytes and other determinants of semen quality?
Eggert-Kruse W, Zwick EM, Batschulat K, Rohr G, Armbruster FP, Petzoldt D, Strowitzki T.
OBJECTIVE: To evaluate a potential association of zinc levels with seminal leukocytes, the outcome of semen cultures; and semen quality and sperm fertilizing capacity . DESIGN: Prospective study . SETTING: Outpatient infertility clinic of a university hospital . PATIENT(S): Two hundred fifty-six randomly chosen asymptomatic males from subfertile couples . INTERVENTION(S): None . MAIN OUTCOME MEASURE(S): Determination of zinc in seminal plasma by flame atomic absorption spectroscopy . In aliquots of the same ejaculates the following tests were performed: immunocytochemical round cell differentiation to determine leukocyte counts and ratios, microbial screening, and comprehensive evaluation of semen quality (sperm analysis, biochemical parameters, antisperm antibody testing, and in vitro examination of sperm ability to penetrate cervical mucus) . The patients underwent medical history, clinical examination, and postcoital testing . Subsequent fertility was determined (controlled for female infertility factors) . RESULT(S): The concentration of zinc in seminal plasma did not correlate in a statistically significant way with leukocytes in semen, nor was it associated with bacterial colonization . There was no statistically significant relationship of zinc in seminal plasma or serum with semen quality parameters nor with local antisperm antibody testing of the IgG or IgA class . Zinc levels did not influence sperm capacity to penetrate cervical mucus in vitro or in vivo, and did not affect subsequent fertility . CONCLUSION(S): The zinc level in seminal fluid and serum is not associated with silent male genital tract infection (indicated by seminal leukocytes); nor is it related to semen cultures in asymptomatic individuals . The lack of association with other semen quality parameters indicates that the routine determination of zinc levels during infertility investigation is not recommended.

Org Lett, 2002 Feb 7, 4(3), 371 - 3
Microbial deracemization of alpha-substituted carboxylic acids; Kato D et al.; An enzyme system of Nocardia diaphanozonaria JCM 3208 catalyzes the inversion of the chirality of various alpha-substituted carboxylic acids, such as 2-phenylpropanoic acid and 2-phenoxypropanoic acid derivatives, via a novel deracemization reaction.

J Contam Hydrol, 2001 Dec 15, 53(3-4), 429 - 53
An analytical quantification of mass fluxes and natural attenuation rate constants at a former gasworks site; Bockelmann A et al.; A new integral groundwater investigation approach was used for the first time to quantify natural attenuation rates at field scale . In this approach, pumping wells positioned along two control planes were operated at distances of 140 and 280 m downstream of a contaminant source zone at a former gasworks site polluted with BTEX- (benzene, toluene, ethyl-benzene, o-, p-xylene) and PAH- (polycyclic aromatic hydrocarbons) compounds . Based on the quantified changes in total contaminant mass fluxes between the control planes, first-order natural attenuation rate constants could be estimated . For BTEX-compounds, these ranged from 1.4e-02 to 1.3e-01 day(-1) whereas for PAH-compounds natural attenuation rate constants of 3.7e-04 to 3.1e-02 day(-1) were observed . Microbial degradation activity at the site was indicated by an increase in dissolved iron mass flux and a reduction in sulphate mass flux between the two investigated control planes . In addition to information about total contaminant mass fluxes and average concentrations, an analysis of the concentration-time series measured at the control planes also allowed to semi-quantitatively delineate the aquifer regions most likely contaminated by the BTEX- and PAH-compounds.

J Contam Hydrol, 2001 Dec 15, 53(3-4), 319 - 40
Microcosm studies of microbial degradation in a coal tar distillate plume; Harrison I et al.; Investigation of a groundwater plume containing up to 24 g l(-1) phenolic compounds suggested that over a period of nearly 50 years, little degradation had occurred despite the presence of a microbial community and electron acceptors within the core of the plume . In order to study the effect of contaminant concentration on degradation behaviour, laboratory microcosm experiments were performed under aerobic and anaerobic conditions at four different concentrations obtained by diluting contaminated with uncontaminated groundwater . The microcosms contained groundwater with total phenols at ca . 200, 250, 660 and 5000 mg l(-1), and aquifer sediment that had been acclimatised within the plume for several months . The microcosms were operated for a period of 390-400 days along with sterile controls to ascertain whether degradation was microbially mediated or abiotic . Under aerobic conditions, degradation only occurred at concentrations up to 660 mg l(-1) total phenols . At phenol concentrations below 250 mg l(-1) a benzoquinone intermediate, thought to originate from the degradation of 2,5-dimethylphenol, was isolated and identified . This suggested an unusual degradative pathway for this compound; its aerobic degradation more commonly proceeding via catecholic intermediates . Under anaerobic conditions, degradation only occurred in the most dilute microcosm (total phenols 195 mg l(-1)) with a loss of p-cresol accompanied by a nonstoichiometric decrease in nitrate and sulphate . By inference, iron(III) from the sediment may also have been used as a terminal electron acceptor, in which case the amount of biologically available iron released was calculated as 1.07 mg Fe(III)/g of sediment . The study shows that natural attenuation is likely to be stimulated by dilution of the plume.

Int Microbiol, 2001 Sep, 4(3), 151 - 7
Ciliate cryptobiosis: a microbial strategy against environmental starvation; Gutierrez JC et al.; This review outlines the main features of ciliate resting-cyst formation or encystment . It represents a strategy against several environmental stresses (such as starvation), which involves a highly gene-regulated cell differentiation process and originates a more resistant, differentiated form or resting cyst . This process is mainly characterized by drastic cytoplasmic dehydration that induces a general metabolic rate decrease, intense autophagic activity, the formation of a permeable cyst wall protecting the cell against the adverse environmental conditions, and a gene-silencing mechanism after opening the specific encystment genes.

Int J Food Sci Nutr, 2002 Jan, 53(1), 29 - 34
Effect of exogenous histidine and Garcinia cambogia on histamine formation in skipjack (Katsuwonus pelamis) homogenates; Thadhani VM et al.; Histamine consumed with food gives rise to allergic reactions . Dark muscle fish, for example skipjack (Katsuwonus pelamis) has been shown to contain histamine . Studies using TLC (acetone: NH4OH, 80:20.5) on silica gel G60 plates and densitometry after spraying with ninhydrin, using a computerized densitometer, showed that freshly harvested skipjack has no detectable histamine (detection limit, 50 micrograms.g-1 fish) . However, with time histamine (Rf 0.84) is formed > 1.5 mg.g-1 probably through microbial action . Skipjack contains high levels of free histidine at levels of > 10 mg.g-1 (Rf 0.41) but fish like seer (Scomberamous spp.), which are not reported to be allergenic, contain < 4 mg.g-1 histidine . Addition of exogenous histidine (50 mg.g-1) results in histamine formation in seer 2.2-fold that of skipjack under the same conditions . A type of herring (Amblygaster spp., sinhala-hurulla) is not a histamine former, but had been shown to cause allergenic reactions, resulting in a ninhydrin positive spot (Rf 0.79) on incubating for 24 hours . Addition of arginine and lysine to blended skipjack results in their loss probably by decarboxylation . Addition of the spice Garcinia cambogia (extracts 0.2 g ml-1) known as 'goraka' in Sri Lanka (sinhala) and 'kukum' in India (hindhi), to fresh skipjack incubates prevents histamine formation as a results of lowering pH to 3.2-3.6 whereas Avverhoea bilimbi (bilin) and Tamarindus indica (tamarind) extracts did not prevent histamine formation.

Swiss Med Wkly, 2001 Oct 20, 131(41-42), 595 - 602
Procalcitonin: how a hormone became a marker and mediator of sepsis; Muller B et al.; Calcitonin was discovered in the early 1960s {1}, at which time it was assumed to be a single hormone with a yet-to-be-determined role in human physiology . Since then it has been found to be only one entity among a large array of related circulating peptides, at least one of which has a pivotal role in the host response to microbial infections {2, 3} . The aim of this review is to describe this metamorphosis of an endocrine hormone to a new class of hormokine mediators in infectious diseases.

Clin Perinatol, 2001 Dec, 28(4), 735 - 51
Intra-amniotic infection in patients with preterm prelabor rupture of membranes . Pathophysiology, detection, and management; Asrat T; The recent increase in knowledge about infection and preterm delivery has engendered many new questions and should make us rethink our long held beliefs and management strategies . Although this article focused primarily on infection as an important factor in the pathogenesis of PPROM, multiple other causes do exist . The various serologic and amniotic fluid assays that can identify activation of the host immune and inflammatory responses as a consequence of the microbial invasion of the amniotic cavity detailed in the preceding paragraphs are very promising but not yet available for clinical use . These tests identify the fetus in the early stages of an infectious process, before the full clinical manifestations of chorioamnionitis . Should such fetuses be treated with antibiotics in an effort to sterilize the amniotic cavity? Should patients with documented microbial invasion be delivered immediately or is there room for a more conservative management with aggressive antibiotic in utero treatment, altering the natural course of PPROM, avoiding extremely preterm deliveries? Certainly many questions remain unanswered . Continuing the search for information on the relationship between infection and PPROM can only add hope to one day finding an option for prevention, because many and probably most cases of PPROM are apparently caused by infection, and the opportunity for preventing this problem most probably lies here.

Antonie Van Leeuwenhoek, 2001 Sep, 79(3-4), 399 - 405
Rare genera of actinomycetes as potential producers of new antibiotics; Lazzarini A et al.; A literature survey covering more than twenty-three thousand bioactive microbial products including eight thousand antiinfectives demonstrated the increasing relevance of the so called 'rare' actinomycetes as a source of new antibiotics . Past and present efforts in the isolation of rare actinomycetes have enriched the Biosearch Italia Strain Collection with more than twenty thousand strains, showing that, when selective isolation methods are developed and extensively applied, some genera, such as Actinomadura, Actinoplanes, Micromonospora, Microtetraspora, are not rare at all and can be recovered from many soil samples . The current focus is on the isolation of members of Streptosporangiaceae family, given their promising chemical diversity.

Adv Biochem Eng Biotechnol, 2001, 73, 129 - 69
Evolutionary engineering of industrially important microbial phenotypes; Sauer U; The tremendous complexity of dynamic interactions in cellular systems often impedes practical applications of metabolic engineering that are largely based on available molecular or functional knowledge . In contrast, evolutionary engineering follows nature's 'engineering' principle by variation and selection . Thus, it is a complementary strategy that offers compelling scientific and applied advantages for strain development and process optimization, provided a desired phenotype is amenable to direct or indirect selection . In addition to simple empirical strain development by random mutation and direct selection on plates, evolutionary engineering also encompasses recombination and continuous evolution of large populations over many generations . Two distinct evolutionary engineering applications are likely to gain more relevance in the future: first, as an integral component in metabolic engineering of strains with improved phenotypes, and second, to elucidate the molecular basis of desired phenotypes for subsequent transfer to other hosts . The latter will profit from the broader availability of recently developed methodologies for global response analysis at the genetic and metabolic level . These methodologies facilitate identification of the molecular basis of evolved phenotypes . It is anticipated that, together with novel analytical techniques, bioinformatics, and computer modeling of cellular functions and activities, evolutionary engineering is likely to find its place in the metabolic engineer's toolbox for research and strain development . This review presents evolutionary engineering of whole cells as an emerging methodology that draws on the latest advances from a wide range of scientific and technical disciplines.

J Antimicrob Chemother, 2002 Feb, 49(2), 243 - 53
The 2001 Garrod lecture . The treatment of cytomegalovirus infection; Griffiths PD; This named lecture provides an opportunity to take an historical perspective on cytomegalovirus (CMV) infection . A major theme will be that modern molecular biological research has questioned the conventional wisdom that CMV is a slow-growing virus, which only damages a few individuals . I will first review details of the genetic constitution of the virus, emphasizing that wild strains contain many genes which are missing from their laboratory-adapted cousins . I will then review the diseases associated with CMV, not just the end-organ diseases of pneumonitis/retinitis, etc., but the so-called indirect effects, including graft rejection, secondary microbial infections and accelerated atherosclerosis . The urgent need for safe and potent antiviral drugs to prevent these diseases will be considered in two ways: first, the failure of the conventional drug discovery approach; and secondly, the opportunities offered by targeting novel gene functions . The controlled clinical trials performed to date will be summarized, together with suggestions about pharmacodynamic evaluations in the future.

Analyst, 2001 Dec, 126(12), 2103 - 6
The potential of 19F NMR spectroscopy for rapid screening of cell cultures for models of mammalian drug metabolism; Corcoran O et al.; The use of microbial cultures as a complementary model for mammalian drug metabolism has been well established previously . Here is a preliminary investigation into the potential of 19F NMR spectroscopy as a rapid screening tool to quantify the biotransformations of fluorine-containing model drugs . Biotransformations of three model drugs in 48 taxonomically diverse organisms were measured by acquiring 19F NMR spectra at 376 MHz . The presence of fluorine in the molecules allowed rapid, simultaneous detection of over 20 biotransformation products without sample pretreatment, chromatography, mass spectrometric techniques or the use of radiolabelled substrates . The detection limit at 376 MHz using 5 mm NMR tubes was ca . 0.3 microg ml(-1) using a typical analysis time of 20 min per sample . With the recent advent of flow injection NMR technology, analysis time of 5 min could be achieved with less sample . This approach may be used to develop fast small-scale microbial screens for the biosynthesis of metabolite standards and production of novel drug analogues, whilst also having a role in reducing animal experiments needed to identify animal and human metabolites of fluorinated xenobiotics.

Nucleic Acids Res, 2001 Dec 15, 29(24), 5169 - 81
The genetic organization and evolution of the broad host range mercury resistance plasmid pSB102 isolated from a microbial population residing in the rhizosphere of alfalfa; Schneiker S et al.; Employing the biparental exogenous plasmid isolation method, conjugative plasmids conferring mercury resistance were isolated from the microbial community of the rhizosphere of field grown alfalfa plants . Five different plasmids were identified, designated pSB101-pSB105 . One of the plasmids, pSB102, displayed broad host range (bhr) properties for plasmid replication and transfer unrelated to the known incompatibility (Inc) groups of bhr plasmids IncP-1, IncW, IncN and IncA/C . Nucleotide sequence analysis of plasmid pSB102 revealed a size of 55 578 bp . The transfer region of pSB102 was predicted on the basis of sequence similarity to those of other plasmids and included a putative mating pair formation apparatus most closely related to the type IV secretion system encoded on the chromosome of the mammalian pathogen Brucella sp . The region encoding replication and maintenance functions comprised genes exhibiting different degrees of similarity to RepA, KorA, IncC and KorB of bhr plasmids pSa (IncW), pM3 (IncP-9), R751 (IncP-1beta) and RK2 (IncP-1alpha), respectively . The mercury resistance determinants were located on a transposable element of the Tn5053 family designated Tn5718 . No putative functions could be assigned to a quarter of the coding capacity of pSB102 on the basis of comparisons with database entries . The genetic organization of the pSB102 transfer region revealed striking similarities to plasmid pXF51 of the plant pathogen Xylella fastidiosa.

Int J Parasitol, 2002 Feb, 32(2), 159 - 66
Sequencing and analysis of a 63 kb bacterial artificial chromosome insert from the Wolbachia endosymbiont of the human filarial parasite Brugia malayi; Ware J et al.; Wolbachia endosymbiotic bacteria are widespread in filarial nematodes and are directly involved in the immune response of the host . In addition, antibiotics which disrupt Wolbachia interfere with filarial nematode development thus, Wolbachia provide an excellent target for control of filariasis . A 63.1 kb bacterial artificial chromosome insert, from the Wolbachia endosymbiont of the human filarial parasite Brugia malayi, has been sequenced using the New England Biolabs Inc . Genome Priming System() transposition kit in conjunction with primer walking methods . The bacterial artificial chromosome insert contains approximately 57 potential ORFs which have been compared by individual protein BLAST analysis with the 35 published complete microbial genomes in the Comprehensive Microbial Resource database at The Institute for Genomic Research and in the NCBI GenBank database, as well as to data from 22 incomplete genomes from the DOE Joint Genome Institute . Twenty five of the putative ORFs have significant similarity to genes from the alpha-proteobacteria Rickettsia prowazekii, the most closely related completed genome, as well as to the newly sequenced alpha-proteobacteria endosymbiont Sinorhizobium meliloti . The bacterial artificial chromosome insert sequence however has little conserved synteny with the R . prowazekii and S . meliloti genomes . Significant sequence similarity was also found in comparisons with the currently available sequence data from the Wolbachia endosymbiont of Drosophila melanogaster . Analysis of this bacterial artificial chromosome insert provides useful gene density and comparative genomic data that will contribute to whole genome sequencing of Wolbachia from the B . malayi host . This will also lead to a better understanding of the interactions between the endosymbiont and its host and will offer novel approaches and drug targets for elimination of filarial disease.

Int Immunopharmacol, 2002 Feb, 2(2-3), 163 - 72
Pulmonary alterations associated with inhalation of occupational and environmental irritants; Castranova V et al.; Many gases, vapors, or particles found in occupational and/or environmental settings can act as irritants . In the present study, sensory irritants are characterized by the stimulation of neuropeptide release from sensory nerves in the nasal mucosa, while pulmonary irritants are characterized by recruitment of PMN into bronchoalveolar airspaces, elevation of breathing frequency, and neuropeptide release from sensory fibers innervating the epithelium of the conducting airways . A review of data from our laboratory as well as results from others indicate that asphalt fume is a sensory irritant; toluene diisocyanate (TDI), methyl isocyanate, and machining fluid act as both sensory and pulmonary irritants; while cotton dust, agricultural dusts, microbial products, leather conditioner, and ozone exhibit responses characteristic of pulmonary irritants.

J Periodontol, 2001 Dec, 72(12), 1652 - 7
Internal contamination of a 2-component implant system after occlusal loading and provisionally luted reconstruction with or without a washer device; Rimondini L et al.; BACKGROUND: Microbial internal contamination of oral implants has been suggested as being responsible for inflammatory infiltration of peri-implant tissues at the fixture-abutment interface of 2-stage and 2-component implant systems; it is also considered as a potential source of pathogens . The present in vivo study evaluated contamination of the inner abutment-implant retaining screw, after occlusal loading and provisionally-luted reconstruction with or without a washer device . METHODS: Eight 2-component oral implants sealed with an o-ring silicon washer device and 9 without a seal were placed in 7 patients with high oral hygiene standards . Two months after prosthetic reconstruction, crowns and internal screws were removed, and organic and inorganic screw contamination was examined by scanning electron microscopy and energy dispersive x-ray spectroscopy (EDS) analysis . RESULTS: An amorphous and crystalline contamination, suggestive of calcium and phosphate compounds, was seen on all screw surfaces . Microbial contamination was more frequently observed in the unsealed group . No differences in bacterial morphotypes were observed between the sealed and unsealed implants . Cocci were the most representative morphotypes, while rods were seldom seen . CONCLUSIONS: In clinical situations, leakage occurs at the implant-abutment interface, although bacterial contamination is limited in patients with high oral hygiene standards . Contamination may be reduced by using a washer device.

Biotechniques, 2002 Jan, 32(1), 160, 162 - 4, 166-71
Viral binding proteins as antibody surrogates in immunoassays of cytokines; Bai H et al.; Cytokines are pivotal to a balanced innate or cell-mediated immune response, can be indicative of disease progression and/or resolution, and are being evaluated as therapeutics . There is a need to purify and/or to measure key cytokines rapidly with accuracy, precision, and sensitivity . The current assay technologies, which are based on RT-PCR, immunoassays, or bioassays, are limited in their use in the clinic, in particular because of the long time (1-3 h) required to carry out the assays . An alternative approach explored here is the use of pathogen-encoded cytokine-binding proteins, which have Kd in the nanomolar range . It is anticipated that pathogens have evolved binding proteins, antagonists, and/or specific neutralizing phenotypes directed against key signaling and effector molecules involved in the multifaceted host defense system . Thus, by screening the genomes of a wide range of microbial agents, we would expect to find coding sequences for binding proteins for the most important cytokines . Consistent with this view is the identification of poxvirus genes encoding binding activities for TNF type I and type II interferons, interleukin (IL)-1beta, IL-18, and beta-chemokines . These high-affinity receptors have the potential to act as surrogate antibodies in a number of applications in cytokine quantification and purification and could be potentially useful reagents to complement the existing panel of anti-cytokine, monoclonal, polyclonal, or engineered antibodies that are currently available.

J Food Prot, 2002 Jan, 65(1), 50 - 2
Monitoring beef carcass surface microbial contamination with a luminescence-based bacterial phosphatase assay; Kang DH et al.; A commercially available microbial phosphatase test kit (Fast Contamination Indicator; FCI) was evaluated as a rapid method for estimating microbial contamination levels on beef carcass tissues . A set of actual beef carcass surface sample swabs (n = 70) was tested using the assay as a means to rapidly (10 min) monitor carcass swab sample microbial contamination . A regression equation was developed in experiment 1 and tested on an independent population . There was agreement between this assay and the conventional plating method for total aerobic mesophilic bacteria (r = 0.93) . The predicted total mesophilic aerobic bacteria count generated from the fitted regression line (predicted log10 CFU/cm2 = 0.7505 x log10 FCI microbial phosphatase test values + 0.6726) showed a high correlation with actual aerobic mesophilic total counts (r = 0.88) . The FCI test offers a simple and rapid method to estimate microbial contamination levels on beef carcasses.

Environ Pollut, 2002, 116(1), 109 - 21
Potentials and drawbacks of chelate-enhanced phytoremediation of soils; Romkens P et al.; Chelate-enhanced phytoremediation has been proposed as an effective tool for the extraction of heavy metals from soils by plants . However, side-effects related to the addition of chelates, e.g . metal leaching and effects on soil micro-organisms, were usually neglected . Therefore, greenhouse and lysimeter studies were conducted to study the phytoremedation potential of EDGA and citric acid and to evaluate its effects on microbial activity and leaching of Cd, Zn Cu and Pb . Grass, lupine and yellow mustard were grown on a moderately polluted acid (pH 4.5) sandy soil that contained 2 mg kg(-1) Cd and 200 mg kg(-1) Zn . Citric acid appeared to be degraded microbially within a few days after addition which limited its potential for long-lasting remediation studies . EDGA enhanced metal solubility but plant uptake did not increase accordingly . The metal shoot:root ratio increased upon addition of EDGA but it also reduced the net shoot and root biomass production of both lupine and yellow mustard . Bacterial biomass was higher in both the citric and EDGA treated pots but bacterial activity remained unaffected . The number of microbivorous nematodes was greatly reduced upon addition of EDGA which was most likely related to the reduced biomass production and, to a smaller extent, to the changes in the composition of the available food . Furthermore, EDGA enhanced metal leaching in the lysimeter study which could lead to groundwater pollution . To prevent these unwanted side-effects, careful management of phytoremediation methods, therefore, seems necessary.

Gerodontology, 2000 Dec, 17(2), 87 - 90
Oral mucous membrane flora in patients using saliva substitutes; Johansson G et al.; Salivary substitutes are sometimes valuable for elderly people and radiotherapy patients, and may be used indefinitely . It is possible that this change in the ecology may effect the oral flora . OBJECTIVE: To analyse the presence of micro-organisms on oral mucous membranes during use of saliva substitutes . DESIGN: Cross-over single-blind study . SETTING: Clinic for Maxillofacial Surgery, Malmo University Hospital and Department of Oral Microbiology, Malmo University . SUBJECTS: 19 patients with low salivary secretion who had been radiated for cancer in the head and neck region . INTERVENTION: Two saliva substitutes: linseed extract and a carboxymethyl cellulose preparation (Salinum and MAS-84) were used for 3 week periods . MEASUREMENTS: Microbial samples taken, processed and analysed . RESULTS: No differences were observed when comparing baseline values with the results after the saliva substitutes and no significant differences between the use of different agents . CONCLUSION: The study suggests that use of linseed extract and carboxymethyl cellulose preparation during periods of weeks does not influence flora commonly related to caries, periodontitis or infections in the oral mucous membranes.

Arch Microbiol, 2002 Feb, 177(2), 123 - 31 Epub 2001 Dec 01.
Biology of extremophilic and extremotolerant methanotrophs; Trotsenko YA et al.; This review summarizes recent findings on the biology of obligate methanotrophic bacteria living in various extreme environments . By using molecular ecology techniques, it has become clear that obligate methanotrophs are ubiquitous in nature and well adapted to high or low temperature, pH and salinity . The isolation and characterization of pure cultures has led to the discovery of several new genera and species of extremophilic/tolerant methanotrophs . Their major physiological role is participation in the methane cycle and supplying C(1) intermediates and various metabolites to other members of microbial communities in extreme ecosystems . To survive under extreme conditions, methanotrophs have developed diverse structure-function adaptive mechanisms including cell-surface layer formation, changes in cellular phospholipid composition and de novo synthesis of organic osmolytes such as ectoine, 5-oxoproline and sucrose . However, despite the above advances, basic knowledge of other stress protectants, as well as bioenergetic and genetic aspects of methanotroph adaptation, is still lacking . This information is necessary for better understanding the molecular mechanisms underlying the versatility of methanotrophs and for the development of novel biotechnological processes.

Environ Pollut, 2002, 116(2), 293 - 300
Plant accumulation of potentially toxic elements in sewage sludge as affected by soil organic matter level and mycorrhizal fungi; Oudeh M et al.; Leek (Allium ameloprasum) was grown in pot trials in two clay loams of contrasting organic contents, with and without indigenous mycorrhizal propagules . Sewage sludges containing varying levels of Cd, Cu and Zn were added . Extractable soil metals, plant growth, major nutrient content and accumulation of metals, and soil microbial indices were investigated . The aim was to establish whether soil organic content and mycorrhizal status affected plant and microbial exposure to these metals . Extractable metals were higher and responses to inputs more pronounced in the arable, lower organic matter soil, although only Cd showed a soil difference in the CaCl2 fraction . There were no metal toxic effects on plants and some evidence to suggest that they promoted growth . Uptake of each metal was higher in the larger plants of the grassland, higher organic matter soil . Inoculation with arbuscular mycorrhizal fungi increased root Cd and Zn concentrations . With the exception of Cd (roots) and Zn (shoots), higher inputs of sludge metals did not increase plant metals . Zn and Cu, but not Cd, concentrations were higher in roots than in shoots.

Cornea, 2002 Jan, 21(1), 33 - 7
Lamellar keratoplasty for the treatment of fungal keratitis; Xie L et al.; PURPOSE: To determine the therapeutic value of lamellar keratoplasty (LKP) in the treatment of fungal keratitis not curable by antifungal chemotherapy . METHODS: Fifty-five patients, in whom a diagnosis of fungal keratitis was confirmed by microscopic analysis of corneal scrapings or confocal microscopy, and who were not cured by topical and oral antifungal medication, were given LKP . After LKP, topical antifungal treatment was continued for 2 weeks with gradual tapering of the drugs . The excised recipient lamella was used for microbial culture and histopathologic examination . RESULTS: Therapeutically beneficial results were achieved in 51 cases (92.7%) of the 55 LKPs that were performed . In these 51 cases, there was no recurrence of infection, and the resulting visual acuity ranged from 20/63 to 20/20 . Patient follow-up ranged from 6 to 18 months . In four cases (7.3%), there was a recurrence of the fungal infection within 2 weeks of LKP . In these four patients, the infection was cured by performing a penetrating keratoplasty (PKP) . Forty-six of the recipient lamellae were culture positive for fungi . Thirty-three of these cultures were identified as Fusarium, six as Aspergillus, three as Candida, one as Penicillium species, and in the other three cases, unidentified septate hyphae were noted . In the four cases of recurrent infection, microbiologic culture revealed three cases with Fusarium species and one case with Aspergillus species . Histopathologic analysis of periodic acid-Schiff (PAS)-stained tissue sections of donor lamellae revealed fungal filaments in all samples . Immune reactions to the lamellar grafts were not observed and the donor lamellae remained clear for the duration of follow-up . CONCLUSION: Lamellar keratoplasty can be effective for treating fungal keratitis that is not cured by antifungal therapeutics . In addition, LKP can provide useful vision with few complications . Furthermore, corneal tissue used in LKP may be obtained more easily than healthy tissue used in PKP.

Proc Natl Acad Sci U S A, 2002 Jan 22, 99(2), 972 - 7
Stereotyped and specific gene expression programs in human innate immune responses to bacteria; Boldrick JC et al.; The innate immune response is crucial for defense against microbial pathogens . To investigate the molecular choreography of this response, we carried out a systematic examination of the gene expression program in human peripheral blood mononuclear cells responding to bacteria and bacterial products . We found a remarkably stereotyped program of gene expression induced by bacterial lipopolysaccharide and diverse killed bacteria . An intricately choreographed expression program devoted to communication between cells was a prominent feature of the response . Other features suggested a molecular program for commitment of antigen-presenting cells to antigens captured in the context of bacterial infection . Despite the striking similarities, there were qualitative and quantitative differences in the responses to different bacteria . Modulation of this host-response program by bacterial virulence mechanisms was an important source of variation in the response to different bacteria.

J Agric Food Chem, 2002 Jan 30, 50(3), 499 - 505
Metabolism of metolachlor by fungal cultures; Sanyal D et al.; Metabolism of metolachlor was studied using a mixed fungal culture isolated from a metolachlor-acclimated field soil . The culture rapidly degraded metolachlor with a half-life of 3.5 days in broth . Aspergillus flavus and A . terricola purified from the mixed culture also metabolized metolachlor effectively . Five metabolites obtained were identified by co-chromatography on HPLC by comparing with authentic standards and by GC-MS . Hydrolytic dechlorination, N-dealkylation, and amide bond cleavage appeared to be the dominant transformations involved in the metabolism . Metabolites, 6-methyl 2-ethyl acetanilide and 6-methyl 2-ethyl aniline, identified in this study are new metabolites of metolachlor being reported from any mixed or pure microbial cultures . The mixed culture could degrade 99% of metolachlor at a fortification level as high as 100 microg mL(-)(1).

Mol Cells, 2001 Dec 31, 12(3), 353 - 9
Tissue specific and inducible expression of resveratrol synthase gene in peanut plants; Chung IM et al.; The expression of resveratrol synthase (RS) genes is induced by biotic and abiotic factors in peanut cell cultures . However, little is known about the regulation of the RS gene expression in peanut plants . The expression of RS genes was investigated in peanut plants with a peanut RS clone, pPRS3C, which encodes two polypeptides that show about a 96% amino acid sequence identity to peanut RS2 and RS3, respectively . A low level of RS mRNA was detected in the roots of peanut plants grown aseptically in vitro . In mature peanut plants that were grown in the field, however, RS mRNAs were present at relatively high levels in both the roots and pods, but at below the detection limit in leaves . RS mRNAs were abundant in young pods and decreased dramatically in mature pods . The RS mRNA expression was induced by yeast extract and UV in leaves and roots, and also by wounding in leaves . Stress hormones, such as ethylene, jasmonic acid, and salicylic acid, induced RS mRNA accumulation in leaves . These results indicate that the RS gene expression is induced by biotic and abiotic stresses through the stress hormones in peanut plants . The induction of the RS gene expression by biotic and abiotic stresses could provide peanut plants with protection from microbial infections through resveratrol synthesis . The RS gene expression in developing pods has significant implications in terms of the role of resveratrol as a phytochemical for human health.

Biomaterials, 2002 Mar, 23(5), 1391 - 7
Effect of surface treatment on the biocompatibility of microbial polyhydroxyalkanoates; Yang X et al.; The biocompatibility of microbial polyesters polyhydroxybutyrate (PHB) and poly(hydroxybutyrate-co-hydroxyhexanoate) (PHBHHx) were evaluated in vitro . The mouse fibroblast cell line L929 was inoculated on films made of PHB, PHBHHx and their blends, polylactic acid (PLA) as control . It was found that the growth of the cells L929 was poor on PHB and PLA films . The viable cell number ranged from 8.8 x 10(2) to 1.8 x 10(4)/cm2 only . Cell growth on the films made by blending PHB and PHBHHx showed a dramatic improvement . The viable cell number observed increased from 9.7 x 10(2) to 1.9 x 10(5) on a series of PHB/PHBHHx blended film in ratios of 0.9/0.1:0/1, respectively, indicating a much better biocompatibility in the blends contributed by PHBHHx . Biocompatibility was also strongly improved when these polymers were treated with lipases and NaOH, respectively . However, the effects of treatment were weakened when PHBHHx content increased in the blends . It was found that lipase treatment had more increased biocompatibility than NaOH . After the treatment biocompatibility of PHB was approximately the same as PLA, while PHBHHx and its dominant blends showed improved biocompatibility compared to PLA.

Water Sci Technol, 2001, 44(11-12), 1 - 8
Fundamental processes within natural and constructed wetland ecosystems: short-term versus long-term objectives; Wetzel RG; Use of wetland ecosystems for water pollution control consists essentially of sustained induced disturbances as pollutants are loaded to complex biological communities . Objectives are to maximize pollutant loading, incorporation, and retention while maintaining highest levels of community metabolism and minimal alteration of community structure . Several basic processes are emphasized: (a) macrophyte productivity in relation to shoot:root ratios, and nutrient availability; (b) macrophyte life history strategies, succession, and biodiversity under constant pollutant stress; (c) importance of standing dead and particulate detritus; (d) functions and controlling mechanisms of heterotrophic and autotrophic periphyton in pollutant retention and recycling; (e) coupling of microbial metabolism to macrophyte retention of pollutants; (f) gaseous losses to the atmosphere; (g) losses of dissolved organic matter and its utilization; and (h) water losses by evapotranspiration and effects on wetland efficacy . Short-term wetland removal efficiencies are confounded by massive variations in retentive capacities diurnally, seasonally, and spatially, in exceeding physiological tolerance levels, and in species succession . Problems of channelization, altered microhydrology, and assimilation/retention are major in natural and non-engineered ecosystems . Wetlands are highly ephemeral and variable in their capabilities for sequestering and retention of nutrients and other pollutants.

Ground Water, 2002 Jan-Feb, 40(1), 76 - 84
Fluid flow and solute migration within the capillary fringe; Silliman SE et al.; Laboratory experiments involving both homogeneous and heterogeneous porous media are used to demonstrate that fluid flow and solute transport will occur regularly in the capillary fringe (CF), including both vertical (upward as well as downward) and horizontal flow velocities . Horizontal flow above the water table appears to be limited primarily to the region of high water saturation (i.e., the CF), an observation supported by numerical modeling and consistent with the literature . Beyond observations presented in prior literature, it was observed that exchange of water within the CF with water below the water table is active, with flux both from the CF downward across the water table and from the region below the water table, upward into the CF . This flux is enhanced by the presence of physical heterogeneity . These findings strongly contrast the common conceptualization of predominantly downward vertical fluid flow through the unsaturated zone, with transition to fully three-dimensional flow only below the water table . Based on these observations, it is suggested that the CF may affect, far more significantly than is usually assumed, the natural geochemical and microbial conditions present in the region of transition from unsaturated to saturated ground water flow.

Metab Eng, 2002 Jan, 4(1), 67 - 79
Manipulating gene expression for the metabolic engineering of plants; Lessard PA et al.; Introducing and expressing foreign genes in plants present many technical challenges that are not encountered with microbial systems . This review addresses the variety of issues that must be considered and the variety of options that are available, in terms of choosing transformation systems and designing recombinant transgenes to ensure appropriate expression in plant cells . Tissue specificity and proper developmental regulation, as well as proper subcellular localization of products, must be dealt with for successful metabolic engineering in plants..

Nat Genet, 2002 Feb, 30(2), 194 - 200 Epub 2002 Jan 22.
Microsatellites are preferentially associated with nonrepetitive DNA in plant genomes; Morgante M et al.; Microsatellites are a ubiquitous class of simple repetitive DNA sequence . An excess of such repetitive tracts has been described in all eukaryotes analyzed and is thought to result from the mutational effects of replication slippage . Large-scale genomic and EST sequencing provides the opportunity to evaluate the abundance and relative distribution of microsatellites between transcribed and nontranscribed regions and the relationship of these features to haploid genome size . Although this has been studied in microbial and animal genomes, information in plants is limited . We assessed microsatellite frequency in plant species with a 50-fold range in genome size that is mostly attributable to the recent amplification of repetitive DNA . Among species, the overall frequency of microsatellites was inversely related to genome size and to the proportion of repetitive DNA but remained constant in the transcribed portion of the genome . This indicates that most microsatellites reside in regions pre-dating the recent genome expansion in many plants . The microsatellite frequency was higher in transcribed regions, especially in the untranslated portions, than in genomic DNA . Contrary to previous reports suggesting a preferential mechanism for the origin of microsatellites from repetitive DNA in both animals and plants, our findings show a significant association with the low-copy fraction of plant genomes.

J Interferon Cytokine Res, 2001 Dec, 21(12), 993 - 1010
Cytokines of birds: conserved functions--a largely different look; Staeheli P et al.; Targeted disruptions of the mouse genes for cytokines, cytokine receptors, or components of cytokine signaling cascades convincingly revealed the important roles of these molecules in immunologic processes . Cytokines are used at present as drugs to fight chronic microbial infections and cancer in humans, and they are being evaluated as immune response modifiers to improve vaccines . Until recently, only a few avian cytokines have been characterized, and potential applications thus have remained limited to mammals . Classic approaches to identify cytokine genes in birds proved difficult because sequence conservation is generally low . As new technology and high throughput sequencing became available, this situation changed quickly . We review here recent work that led to the identification of genes for the avian homologs of interferon-alpha/beta (IFN-alpha/beta) and IFN-gamma, various interleukins (IL), and several chemokines . From the initial data on the biochemical properties of these molecules, a picture is emerging that shows that avian and mammalian cytokines may perform similar tasks, although their primary structures in most cases are remarkably different.

J Infect, 2001 Oct, 43(3), 182 - 6
Risk of infection and death among post-splenectomy patients; Bisharat N et al.; OBJECTIVES: The true incidence of post-splenectomy sepsis remains undetermined . METHODS: An English literature review on post-splenectomy sepsis was undertaken by means of databases of MEDLINE for the period 1966-96 . The data registered included age at splenectomy, indication for splenectomy, incidence of infection and death, interval between splenectomy and infection, and microbial aetiology . RESULTS: The reports include 19 680 patients having undergone splenectomy with a median follow up of 6.9 years . The incidence of infection after splenectomy was 3.2% and the mortality rate was 1.4% . Only 6942 reports were sufficiently detailed to allow useful analysis . The incidence of infection among children and adults was similar, 3.3% and 3.2%, respectively . However, the death rates among children were higher than adults (1.7% vs . 1.3%) . The incidence of infection was highest among patients with thalassemia major (8.2%), and sickle-cell anaemia (7.3%) . The highest mortality rates were observed among patients with thalassaemia major (5.1%), and sickle-cell anaemia (4.8%) . CONCLUSION: The incidence of sepsis among post-splenectomy patients is low, however, it carries a high mortality rate especially among children with hematological disorders .

Sheng Wu Gong Cheng Xue Bao, 2001 Sep, 17(5), 515 - 9
{PCR amplification of anaerobic fungal 18S rDNA from landfill sites}; Hang XM et al.; Nucleic acid based techniques are widely used to characterize microbial communities in environmental samples . In this study, seven landfill leachates were tested for the presence of 18S rRNA genes from anaerobic fungi from the family Chytridiomycetes . A nested Polymerase Chain Reaction (PCR) strategy was employed to amplify the 18S rRNA gene region of Chytridiomycetous fungi from landfill sites samples . All of the PCR products from landfill leachates amplified with the first primer pair NS1/NS8 and the second primer pair Chyt-719/Chyt-1553 were subjected to agarose gel electrophoresis to show seven samples containing fungi from the family Chytridiomycetes . One of the positive results was then selected for cloning and sequencing of the PCR product to confirm the identity of the amplified DNA . The molecular data presented here reveals for the first time that Chytridiomycetous fungi associated with the anaerobic rumen environment are also present in landfill sites.

J Urban Health, 2001 Dec, 78(4), 647 - 57
The acceptability of reuse of the female condom among urban South African women; Pettifor AE et al.; This study assessed whether reuse of the female condom was acceptable among two groups of women in central Johannesburg, South Africa, who were taking part in two separate studies of female condom reuse . The first group consisted of women (aged 17 to 43 years) attending a family planning/sexually transmitted infections (STIs) clinic who were participating in a cross-sectional survey of the acceptability of female condoms reuse (n = 100) . The second group included women (aged 18-40 years) at high risk for STI (80% self-declared sex workers) who were taking part in an ongoing cohort study to investigate the safety of reuse of the female condom through a structural integrity and microbial retention study (n = 50) . Among women participating in the acceptability study, 83% said that they would be willing to reuse the female condom, and 91% thought the idea of reuse of the female condom was acceptable . All women taking part in the safety of reuse study and who reused the female condom up to seven times (n = 49) reported that the steps involved in reusing the device were easy to perform and acceptable . All 49 women said they would reuse the female condom at least once, while 45% said they would use it a maximum of seven or eight times . From the results of the interviews with both study groups, it can be concluded that, among women in a South African urban environment who have used a male and/or female condom, the concept of reuse of the female condom is acceptable and thought to be a good idea.

Infect Immun, 2002 Feb, 70(2), 844 - 50
Chemokine and chemokine receptor dynamics during genital chlamydial infection; Belay T et al.; Current design strategies for vaccines against certain microbial pathogens, including Chlamydia trachomatis, require the induction and targeting of specific immune effectors to the local sites of infection known as the mucosal effector sites . Chemokines and their receptors are important mediators of leukocyte trafficking and of the controlled recruitment of specific leukocyte clonotypes during host defense against infections and during inflammation . We analyzed the dynamics of chemokine and chemokine receptor expression in genital mucosae during genital chlamydial infection in a murine model to determine how these molecular entities influence the development of immunity and the clearance of infection . A time course study revealed an increase of up to threefold in the levels of expression of RANTES, monocyte chemotactic protein 1 (MCP-1), gamma-interferon-inducible protein 10 (IP-10), macrophage inflammatory protein 1alpha (MIP-1alpha), and intercellular adhesion molecule type 1 (ICAM-1) after genital infection with the C . trachomatis agent of mouse pneumonitis . Peak levels of expression of RANTES, MCP-1, and MIP-1alpha occurred by day 7 after primary infection, while those of IP-10 and ICAM-1 peaked by day 21 . Expression levels of these molecules decreased by day 42 after primary infection, by which time all animals had resolved the infection, suggesting an infection-driven regulation of expression . A rapid upregulation of expression of these molecules was observed after secondary infection . The presence of cells bearing the chemokine receptors CCR5 and CXCR3, known to be preferentially expressed on Th1 and dendritic cells, was also synchronous with the kinetics of immune induction in the genital tract and clearance of infection . Results demonstrated that genital chlamydial infection is associated with a significant induction of chemokines and chemokine receptors that are involved in the recruitment of Th1 cells into the site of infection . Future studies will focus on how selective modulation of chemokines and their receptors can be used to optimize long-term immunity against CHLAMYDIA:

J Biotechnol, 2002 Mar 14, 94(1), 101 - 23
Xenobiotics in the environment: present and future strategies to obviate the problem of biological persistence; Rieger PG et al.; Sustainable chemistry aims at an improved efficiency of using natural resources which are used to meet human needs for chemical products . Chemists in science and industry, have become aware of the importance to design environmentally benign chemicals . One aspect is the biological persistence and the present paper reviews work in this field focussing on the degradation of xenobiotics in the environment . Different structural reasons for chemical and biological persistence are described and strategies to use single bacterial isolates or microbial communities for the elimination of xenobiotic pollutants in the environment are summarized . Perspectives and limitations to evolve and use this catabolic potential are critically discussed with respect to the complexity of mixtures of xenobiotics often found in practice . An interdisciplinary approach for the prospective design of environmentally benign substances is presented and examples for new commodity chemicals that better fit the naturally existing catabolic potential are included.

Transfus Apheresis Sci, 2001 Aug, 25(1), 43 - 50
Mononuclear cells ingest E . coli opsonized by investigational intravenous immunoglobulin preparations in the absence of complement more efficiently than polymorphonuclear phagocytes; Tinguely C et al.; OBJECTIVE: The increasing use of intravenous immunoglobulins (IVIg) to treat systemic inflammatory, mostly autoimmune, diseases raises the concern that their competition for FcgammaReceptor (FcgammaR) binding with immune complexes might interfere with the removal of opsonized microbial agents more than it favoured their opsonophagocytosis . METHODS: Using a sensitive FACS-based phagocytosis assay allowing for the identification of the phagocytosing cell-type, we have tested the relative capacity of Pentaglobin and two investigational IVIg preparations, IVIgG and IVIgM, to promote E . coli phagocytosis . The plasma IL-8 levels 4 h after phagocytosis were measured by ELISA . RESULTS: Heat inactivated, complement (C) depleted serum allowed about 25% polymorphonuclear leukocytes (PMNLs) and 15% monocytes (MNs) of 16 donors to phagocyte E . coli . The addition of IVIg favoured mean phagocytosis by MNs to 55%, but by PMNLs only to 25% . The addition of fresh autologous plasma as a C source resulted in a boost of phagocytosing capacity to reach >90% of the PMNLs and >72% of the MNs . Blockade of FcgammaRII and FcgammaRI using monoclonal antibodies (mAb) reduced phagocytosis and IL-8 release . CONCLUSION: We conclude that excess IVIgG/M preparations added to a phagocytic system of PMNLs and MNs, without preopsonization of E . coli, promotes phagocytosis as expected, but MNs rather than PMNLs do this more thoroughly on their own.

Genome Inform Ser Workshop Genome Inform, 2001, 12, 63 - 72
Estimation of bidirectional metabolic fluxes from MS and NMR data using positional representations; Arauzo-Bravo MJ et al.; It is quite important to estimate the metabolic flux distribution (MFD) vectors in vivo, and to investigate the effect of culture environments on the flux distributions to uncover the metabolic regulation mechanism of microbial cells . The conventional approach is to compute the MFD using the stoichiometric equations and the measured specific rates (input and output variables) . However, this method cannot give the MFD for the complex metabolic network which includes cyclic pathways . In the present investigation, we considered the method of analysing the metabolic fluxes based on 13C tracer experiments . In particular, we compared the different techniques of estimating the bidirectional fluxes in the metabolic networks, studying their applicability with respect to the different types of data formats obtained through GC-MS (gas chromatography-mass spectrometry) and NMR (nuclear magnetic resonance) measurements in labeling experiments . It was found that some techniques cannot be applied for GC-MS and NMR data . In the present research, therefore, a new preprocessing method for MS and NMR data was developed, to solve some of the problems encountered in the conventional approaches.

Curr Opin Immunol, 2002 Feb, 14(1), 136 - 45
Phagocytosis and innate immunity; Greenberg S et al.; Phagocytosis is an evolutionarily conserved process utilized by many cells to ingest microbial pathogens, and apoptotic and necrotic corpses . Recent investigation has revealed a fundamental requirement for two co-ordinated cellular processes--cytoskeletal alterations and membrane trafficking--in the phagocytic event . Some elements of this machinery are co-opted by certain pathogens to gain entry into host cells.

Curr Opin Immunol, 2002 Feb, 14(1), 96 - 102
Defensins of vertebrate animals; Lehrer RI et al.; During the past year, novel beta-defensins of mice and men have been identified, together with a novel defensin subfamily (the circular or 'theta' minidefensins) in primates . Insight into the evolution of defensins has been obtained from structural studies, and several mechanisms related to microbial resistance to defensins have been delineated . There is now convincing evidence that defensins augment adaptive immune responses.

J Environ Qual, 2001 Nov-Dec, 30(6), 1896 - 903
Methane oxidation and production activity in soils from natural and agricultural ecosystems; Chan AS et al.; Methane (CH4) flux from soil to the atmosphere is the result of two microbial processes, methanogenesis and CH4 oxidation . Land use may have a profound impact on the relative activities of these groups of organisms . In this study, the CH4 production and consumption potentials of soils from agricultural and nonagricultural ecosystems were assessed in laboratory incubations . Methane production potentials of most soils were low and in the range of 0.02 to 0.35 nmol CH4 g soil(-1) h(-1); however, soils from two of the agricultural sites that experience periodic water saturation had CH4 production potentials from 100 to 300 nmol CH4 g soil(-1) h(-1) . The high methanogenic potential suggests that CH4 consumers may not be wholly dependent on atmospheric CH4 for their survival and maintenance . The prairie soils exhibited the highest CH4 oxidation under ambient atmospheric CH4 concentrations, and CH4 oxidation activity was markedly enhanced in incubations with an atmosphere enriched in CH4 . This stimulated CH4 oxidation activity was generally greater in the agricultural soils as compared with the forest and prairie soils . Methane oxidation appeared to be related to soil nitrogen status . Under ambient atmospheric CH4 concentrations, CH4 oxidation was negatively related to soil mineral N (NO2- + NO3- + NH4+) concentration . However, a positive relationship between soil mineral N status and CH4 oxidation activity was observed in incubations with atmospheres enriched in CH4 . This pattern suggests that the agricultural lands contain different populations of CH4 oxidizers than the natural systems.

Vasc Med, 2001, 6(3), 145 - 9
Plasma levels of cystatin-C and mannose binding protein are not associated with risk of developing systemic atherosclerosis; Albert MA et al.; Cystatin-C, a cysteine protease inhibitor, and mannose binding lectin, an innate defense protein involved in microbial clearance, have both been hypothesized to mediate atherosclerotic plaque progression . Prospective data evaluating whether levels of these proteins are associated with incident cardiovascular disease are sparse . Employing a prospective, nested, case-control study design, baseline levels of cystatin-C and mannose binding protein were evaluated among 133 apparently healthy men who subsequently developed symptomatic peripheral arterial disease (cases) and among 133 age- and smoking-matched controls who remained free of reported vascular disease during 5 years of follow-up . Overall, median baseline levels of cystatin-C were virtually identical among case and control subjects (0.83 mg/l, p = 0.84), whereas median baseline levels of mannose binding protein among cases and controls were 2.32 mg/l and 2.20 mg/l respectively (p = 0.69) . No evidence of association was found between either cystatin-C or mannose binding protein and the development of peripheral arterial disease in analyses evaluating for linear trends or for threshold effects (all p-values > 0.05) . In contrast with prior retrospective and cross-sectional studies, no evidence was found that baseline levels of cystatin-C or mannose binding protein are associated with an increased risk of future arterial disease.

Reprod Nutr Dev, 2001 Jul-Aug, 41(4), 349 - 64
Effect of level and origin of rumen degradable nitrogen on rumen microbial growth and nitrogen utilisation efficiency of animals fed maize silage at maintenance; Fievez V et al.; Isoenergetic maize silage diets, fed at maintenance to 24 suckling cows (Exp . 1) and to 3 (Exp . 2) or 4 (Exp . 3) rumen fistulated sheep, were compared with a urea and controlled release NPN diet (Exps . 1 and 2) and with a protein-N supplemented diet (Exp . 3) . Supplementation increased blood urea concentrations (44.7 +/- 22.3 vs . 97.6 +/- 23.7 mg urea-N x L(-1)) (Exp . 1) and renal urea excretion (2.5 +/- 1.1 vs . 7.6 +/- 1.8 g urea-N x d(-1)) (Exps . 2 and 3), whereas blood allantoin concentrations (286.7 +/- 77.0 micromol x L(-1)) (Exp . 1) and renal excretion of purine derivatives (357.6 +/- 90.7 mg purine-N x d(-1)) (Exps . 2 and 3) were not affected, indicating additional N supplementation did not improve rumen microbial growth . However, some deficiency of rumen degradable N might occur in non supplemented diets as suggested by the reduced rumen NH3-N and RNA concentrations (868 +/- 270 vs . 1466 +/- 466 mg RNA x kg(-1) rumen contents).

Int Dent J, 2001 Dec, 51(6), 417 - 27
General health risk of periodontal disease; Slots J et al.; The possibility that periodontal disease might influence the morbidity and mortality of systemic diseases constitutes a research topic of great current interest . Human periodontal disease is associated with a complex microbiota containing approximately 500 microbial taxa and various human viruses, many of which possess significant virulence potential . Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and other periodontopathic bacteria that are unique to the oral cavity and may disseminate to other body sites comprise the best-documented form of dental focal infection . However, systemically healthy individuals seem to be at low risk of acquiring acute non-oral diseases from direct infections by periodontal pathogens . Research data from various laboratories point to periodontal infections as a risk factor for chronic medical disorders, including cardiovascular disease, cerebrovascular accidents and low-birth-weight infants . However, recent epidemiological studies have failed to show a significant relationship between periodontal disease and cardiovascular disease . This review paper evaluates the current status of knowledge on dental focal infection and suggests avenues for further research into the topic of general health risks of periodontal disease.

Nat Genet, 2002 Feb, 30(2), 141 - 2 Epub 2002 Jan 14.
Identification of foreign gene sequences by transcript filtering against the human genome; Weber G et al.; We have developed a computational subtraction approach to detect microbial causes for putative infectious diseases by filtering a set of human tissue-derived sequences against the human genome . We demonstrate the potential of this method by identifying sequences from known pathogens in established expressed-sequence tag libraries.

J Trace Elem Med Biol, 2001, 15(2-3), 109 - 14
The influence of dietary microbial phytase and calcium on the accumulation of cadmium in different organs of pigs; Zacharias B et al.; A total of 72 barrows (initial body weight 16.7 kg) was used, to evaluate the influence of microbial phytase supplementation alone or in combination with calcium to barley soybean meal diets on the accumulation of cadmium (Cd) in kidney, liver, muscle, brain and bone . The control group received the basal diet with 6 g Ca and a low native Cd concentration of 0.03 mg/kg dry matter (DM) . In the experimental groups 2, 3, 4 and 5 dietary cadmium concentration was elevated to 0.78 mg/kg DM . The diet of group 3 was supplemented with 800 U microbial phytase/kg, the diet of group 4 with 6 g Ca/kg . The diet of group 5 contained both supplements . The addition of microbial phytase caused an increase of Cd retention in kidney and liver at 30 and 50 kg body weight . This effect was counteracted by the contemporary addition of calcium . A supplementation of Ca alone showed no effect on the Cd accumulation in kidney and liver . In muscle, brain and bone no effects of phytase and calcium on the accumuLation of Cd could be found.

Biotechnol Bioeng, 2002 Feb 15, 77(4), 467 - 75
Quantitative determination of cellulase concentration as distinct from cell concentration in studies of microbial cellulose utilization: analytical framework and methodological approach; Lynd LR et al.; In analyzing microbial cellulose utilization, it would be useful to independently measure the mass concentration of cells and cellulase enzymes . Such measurements would allow investigation of the allocation of cellular resources between synthesis of cells and cellulase, in vivo cell- and cellulase-specific cellulose hydrolysis rates, and bioenergetics . Methodological protocols are not established for independent determination of cell and cellulase concentrations for the common case in which a substantial fraction of cellulase is attached to the cell surface . Alternative analytical approaches by which to develop such protocols are examined from the perspective of error minimization . For cell concentration measurement, acceptable accuracy is expected when the concentrations of a cell-specific component (e.g., DNA) is determined or when total protein is determined in conjunction with a measurement specific to cellulase . For cellulase concentration measurement, acceptable accuracy is expected when a measurement specific to cellulase such as ELISA is used . Several analytical approaches are rejected based on large expected errors .

Biotechnol Bioeng, 2002 Feb 15, 77(4), 394 - 404
Development of an ultrahigh-temperature process for the enzymatic hydrolysis of lactose . III . Utilization of two thermostable beta-glycosidases in a continuous ultrafiltration membrane reactor and galacto-oligosaccharide formation under steady-state conditions; Petzelbauer I et al.; Hydrolysis of lactose by hyperthermophilic beta-glycosidases from the archaea Sulfolobus solfataricus (SsbetaGly) and Pyrococcus furiosus (CelB) was carried out at 70 degrees C in a continuous stirred-tank reactor (CSTR) coupled to a 10-kDa cross-flow ultrafiltration module to recycle the enzyme . Recirculation rates of > or =1 min(-1), reaction of proteins with reducing sugars, and enzyme adsorption onto the membrane are major "operational" factors of enzyme inactivation in the CSTR . They cause the half-life times of SsbetaGly and CelB to be reduced two- and eight-fold, respectively, the average value for both enzymes now being approximately 5 to 7 days . Using lactose at initial concentrations of 45 and 170 g/L, the CSTR was operated at a constant conversion level of approximately 80% for more than 2 weeks without the occurrence of microbial contamination . The productivities for the SsbetaGly-catalyzed conversion of lactose were determined at different dilution rates and initial substrate concentrations, and exceed by a factor of < or =2 those observed with CelB under otherwise identical conditions . This difference reflects the approximately eight-fold stronger product inhibition of CelB by D-glucose . While the maximum total galacto-oligosaccharide production (90-100 mM) at 170 g/L lactose in the CSTR was not different from that in the batch reactor (CelB) or was greater by approximately 25% (SsbetaGly), continuous and batchwise reactions with both enzymes differed markedly with regard to relative proportions of the individual saccharide components present at 80% substrate conversion . The CSTR yielded an up to four-fold greater ratio of disaccharides to trisaccharides concomitant with a 5- to 30-fold larger relative proportion of beta-D-Galp-(1-->3)-D-Glc in the product mixture . The results show that apart from continuous hydrolysis of lactose at 70 degrees C, a CSTR charged with SsbetaGly or CelB and operated at steady-state conditions could be a useful reaction system for the production of galacto-oligosaccharides in which composition is narrower and more easily programmable, in terms of the individual components contained, as compared to the batchwise reaction .

Am J Phys Anthropol, 2001, Suppl 33, 54 - 69
Loss of N-glycolylneuraminic acid in humans: Mechanisms, consequences, and implications for hominid evolution; Varki A; The surface of all mammalian cells is covered with a dense and complex array of sugar chains, which are frequently terminated by members of a family of molecules called sialic acids . One particular sialic acid called N-glycolylneuraminic acid (Neu5Gc) is widely expressed on most mammalian tissues, but is not easily detectable on human cells . In fact, it provokes an immune response in adult humans . The human deficiency of Neu5Gc is explained by an inactivating mutation in the gene encoding CMP-N-acetylneuraminic acid hydroxylase, the rate-limiting enzyme in generating Neu5Gc in cells of other mammals . This deficiency also results in an excess of the precursor sialic acid N-acetylneuraminic acid (Neu5Ac) in humans . This mutation appears universal to modern humans, occurred sometime after our last common ancestor with the great apes, and happens to be one of the first known human-great ape genetic differences with an obvious biochemical readout . While the original selection mechanisms and major biological consequences of this human-specific mutation remain uncertain, several interesting clues are currently being pursued . First, there is evidence that the human condition can explain differences in susceptibility or resistance to certain microbial pathogens . Second, the functions of some endogenous receptors for sialic acids in the immune system may be altered by this difference . Third, despite the lack of any obvious alternate pathway for synthesis, Neu5Gc has been reported in human tumors and possibly in human fetal tissues, and traces have even been detected in normal human tissues . One possible explanation is that this represents accumulation of Neu5Gc from dietary sources of animal origin . Finally, a markedly reduced expression of hydroxylase in the brains of other mammals raises the possibility that the human-specific mutation of this enzyme could have played a role in human brain evolution . Yrbk Phys Anthropol 44:54-69, 2001 .

Science, 2002 Jan 11, 295(5553), 338 - 42
Distinct effects of T-bet in TH1 lineage commitment and IFN-gamma production in CD4 and CD8 T cells; Szabo SJ et al.; T-bet is a member of the T-box family of transcription factors that appears to regulate lineage commitment in CD4 T helper (TH) lymphocytes in part by activating the hallmark TH1 cytokine, interferon-gamma (IFN-gamma) . IFN-gamma is also produced by natural killer (NK) cells and most prominently by CD8 cytotoxic T cells, and is vital for the control of microbial pathogens . Although T-bet is expressed in all these cell types, it is required for control of IFN-gamma production in CD4 and NK cells, but not in CD8 cells . This difference is also apparent in the function of these cell subsets . Thus, the regulation of a single cytokine, IFN-gamma, is controlled by distinct transcriptional mechanisms within the T cell lineage.

Am J Pathol, 2002 Jan, 160(1), 165 - 73
Commensal-associated molecular patterns induce selective toll-like receptor-trafficking from apical membrane to cytoplasmic compartments in polarized intestinal epithelium; Cario E et al.; Commensal-associated molecular patterns, the major products of nonpathogenic bacteria, are present at high concentrations at the apical surface of the intestinal epithelium . However, the nature of the interaction of commensal-associated molecular patterns with the lumenal surface of the epithelium has not been defined . We have recently demonstrated that intestinal epithelial cells constitutively express several Toll-like receptors (TLRs) in vitro and in vivo that seem to be the key receptors responsible for immune cell activation in response to various bacterial products . In this study we characterize the subcellular distribution of two major TLRs, TLR2 and TLR4, and their ligand-specific dynamic regulation in the model human intestinal epithelial cell line T84 . Immunocytochemical studies indicate that TLR2 and TLR4 are constitutively expressed at the apical pole of differentiated T84 cells . After stimulation with lipopolysaccharide or peptidoglycan, TLRs selectively traffic to cytoplasmic compartments near the basolateral membrane . Thus, we demonstrate that TLRs are positioned at the apical pole where they are poised to monitor the sensitive balance of the lumenal microbial array . The results of this dynamic epithelial surveillance can then be conveyed to the underlying cell populations of the lamina propria via these innate immune pattern recognition receptors.

Med Sci Monit, 2002 Jan, 8(1), RA1 - 4
Tonal nitric oxide and health--a free radical and a scavenger of free radicals; Benz D et al.; Basal/tonal nitric oxide (NO) production helps maintain particular microenvironments, i.e., vascular . Besides NO's function in controlling the activation state of various tissues such as immune cells, its presence appears to modulate other free radical levels, i.e., H2O2, in these same tissues and indeed these processes may be one and the same . Thus, by being a free radical, along with the ability to scavenge other free radicals, NO is placed in a pivotal regulatory position . We surmise that in the absence of adequate NO release other free radicals may go 'unchecked' and, therefore, initiate tissue damage . Furthermore, under these circumstances, proinflammatory events will occur due to heightened cell sensitivity and a diminished control of NF-kappaB . In an excess situation, and one without an appropriate circumstance, i.e., microbial action, NO may become the harmful agent . Hence, balancing basal NO production in body compartments may represent a fundamental process in maintaining general, long-term health.

J Ind Microbiol Biotechnol, 2001 Sep, 27(3), 136 - 43
Disclosure of new and recurrent microbial metabolites by mass spectrometric methods; Grafe U et al.; The application of modern mass spectrometry methods (SI-CID-MS/MS; MSn) in the disclosure of new and recurrent microbial metabolites is discussed . Spray ion (SI) sources coupled to different kinds of mass analyzers enable the determination of molecular weights and chemical formulas of given samples even in mixtures . Diagnostic fragment formation by collision-induced dissociation (CID-MS/MS) and MSn experiments using ion trap mass analyzers are shown as another indispensable source of structural information . Due to the development of benchtop-type mass spectrometers coupled to high-performance liquid chromatography (HPLC), MS can be practised in almost every laboratory as a powerful tool in natural product analysis . Examples are given for special MS applications in identification of bioactive metabolites from screening strains.

J Dent Educ, 2001 Dec, 65(12), 1335 - 9
A revolution in biomedical assessment: the development of salivary diagnostics; Tabak LA; Since the early 1900s, saliva has proven to be a noninvasive medium from which to measure a wide range of hormones, pharmaceuticals, and antibodies . It has also proven to be a convenient source of host and microbial DNA . As we enter the era of genomic medicine, increasing use of salivary diagnostics will help catalyze a shift from disease diagnosis to health surveillance . However, with the advances in this technology comes the additional obligation to ensure the privacy and rights of patients.

Biochem Biophys Res Commun, 2002 Jan 11, 290(1), 236 - 41
Human lactoferrin activates transcription of IL-1beta gene in mammalian cells; Son KN et al.; Lactoferrin (Lf) has been suggested to play roles in primary defense against microbial infection and other cellular processes including immunomodulation . Lf is known to bind to DNA and implicated to activate transcription . In the present study, we demonstrated that Lf stimulated transcription of IL-1beta gene, one of natural genes containing putative Lf binding site (LBS) in the 5'-flanking sequences . K562 cells treated with a combination of Lf and PMA showed a synergistic induction in the level of IL-1beta mRNA over treatment with PMA alone . Synergistic stimulation of IL-1beta expression by Lf and PMA was also confirmed by IL-1beta/Luc reporter gene assays . Analysis of Lf domains revealed that the transcriptional domain of Lf is located within the N-terminal 90 amino acids, termed NIa and that the C-terminal half lobe lacked the transactivating activity . The NIa, the N-terminal half lobe as well as intact Lf stimulated transcription of IL-1beta gene in the transfected K562 cells along with PMA, while the C-terminal half lobe did not . Our results suggest that Lf may play some roles in transcription of IL-1beta gene and may also regulate transcription of other natural genes containing LBS . (c)2002 Elsevier Science.

Sci Total Environ, 2001 Dec 17, 281(1-3), 111 - 9
210Po and 210Pb cycling in a hydrothermal vent zone in the coastal Aegean sea; Boisson F et al.; To quantify the potential enhancement of naturally-occurring 210Po and 210Pb that may result from the high sulfur-reducing and sulfur-oxidizing regimes associated with hydrothermal vents, sinking particles from both inside and outside vent areas and benthic molluscs grazing on or living near bacterial mats in the vent zone were collected off Milos Island (Aegean Sea) and analyzed for their 210Po and 210Pb content . There was no significant difference in the range of 210Po specific activities measured in particulate material collected by sediment traps in a control area and in the vent area; the resultant 210Po levels were of the same order of magnitude as literature values reported for other Mediterranean coastal areas . 211Pb levels in sinking particles from the control site tended to be higher than those measured in the vent zone, as demonstrated by the lower 210Po/210Pb ratios observed in particles from the control site . Nevertheless, these 210Pb levels were also comparable with similar 210Pb data reported for the northwestern Mediterranean Sea . The 210Po and 210Pb vertical particulate fluxes were, on average, higher in the vent zone as a consequence of the higher particle flux . This observation indicates that vents can indirectly control the flux of these natural radionuclides by affecting the types and amount of particles produced in hydrothermal areas . The 210Po levels measured in a gastropod and a bivalve living on or near the microbial mat in the vent zone were higher than values reported for non-vent gastropods and bivalves from the NW Mediterranean Sea, an observation which suggests that an enhanced food chain transfer of 210Po may occur in the vicinity of vents off Milos Island . Nevertheless, the lack of a general enhancement of 210Po and 210Pb in the marine particulate samples collected indicates that any input of these radionuclides through venting activity may have a minimal effect in the surrounding environment.

J Microbiol Methods, 2002 Mar, 49(1), 19 - 29
A sandwich-designed temperature-gradient incubator for studies of microbial temperature responses; Elsgaard L et al.; A temperature-gradient incubator (TGI) is described, which produces a thermal gradient over 34 aluminium modules (15x30x5 cm) intersected by 2-mm layers of partly insulating graphite foil (SigraFlex Universal) . The new, sandwich-designed TGI has 30 rows of six replicate sample wells for incubation of 28-ml test tubes . An electric plate heats one end of the TGI, and the other end is cooled by thermoelectric Peltier elements in combination with a liquid cooling system . The TGI is equipped with 24 calibrated Pt-100 temperature sensors and insulated by polyurethane plates . A PC-operated SCADA (Supervisory Control And Data Acquisition) software (Genesis 4.20) is applied for temperature control using three advanced control loops . The precision of the TGI temperature measurements was better than +/-0.12 degrees C, and for a 0-40 degrees C gradient, the temperature at the six replicate sample wells varied less than +/-0.04 degrees C . Temperatures measured in incubated water samples closely matched the TGI temperatures, which showed a linear relationship to the sample row number . During operation for 8 days with a gradient of 0-40 degrees C, the temperature at the cold end was stable within +/-0.02 degrees C, while the temperatures at the middle and the warm end were stable within +/-0.08 degrees C (n=2370) . Using the new TGI, it was shown that the fine-scale (1 degrees C) temperature dependence of S(o) oxidation rates in agricultural soil (0-29 degrees C) could be described by the Arrhenius relationship . The apparent activation energy (E(a)) for S(o) oxidation was 79 kJ mol(-1), which corresponded to a temperature coefficient (Q(10)) of 3.1 . These data demonstrated that oxidation of S(o) in soil is strongly temperature-dependent . In conclusion, the new TGI allowed a detailed study of microbial temperature responses as it produced a precise, stable, and certifiable temperature gradient by the new and combined use of sandwich-design, thermoelectric cooling, and advanced control loops . The sandwich-design alone reduced the disadvantageous thermal gradient over individual sample wells by 56%.

J Microbiol Methods, 2002 Feb, 48(2-3), 259 - 65
Identification of bacteria from single colonies by fatty acid analysis; Buyer JS; While a commercially available system for microbial identification by fatty acid analysis (Microbial Identification System, MIDI, Newark, DE, USA) is available, it requires approximately 40-mg wet weight of biomass . Various authors have published methods for fatty acid analysis of single colonies, but no database of organisms has been developed for those methods . A modification of the MIDI system to increase sensitivity while maintaining relative retention times and peak areas allows the standard peak naming tables and libraries of organisms to be used with single colonies . Samples are evaporated down before injection to concentrate the fatty acids, while splitless injection is used to allow more sample to enter the gas chromatograph . Several known bacteria were correctly identified by this method.

J Microbiol Methods, 2002 Feb, 48(2-3), 101 - 6
Laser desorption mass spectrometry for microbial DNA analysis; Taranenko NI et al.; Recently, we demonstrated that a matrix-assisted laser desorption/ionization time-of-flight mass spectrometer (MALDI-TOF MS) can be used to determine the molecular weight of polymerase chain reaction (PCR) products of intact 16S rRNA regions and to profile their restriction digests . This is the first time that MALDI-TOF MS with ultraviolet (UV) photoionization has been used to analyze a PCR product of approximately 1600 nucleotides in length.

J Econ Entomol, 2001 Dec, 94(6), 1419 - 27
Field evaluation of a phototoxic dye, phloxine B, against three species of fruit flies (Diptera: Tephritidae); Moreno DS et al.; The xanthene dye phloxine B (D&C Red #28) bait was sprayed against fruit flies in mango orchards in 1996 and 1997 . The flies used for testing were Mexican fruit fly, Anastrepha ludens (Loew), West Indian fruit fly, Anastrepha obliqua (Macquart), and Mediterranean fruit fly, Ceratitis capitata (Wiedeman) . Results of the experiments indicate that the toxic efficacy of phloxine B against these fruit flies is as good as that of malathion-bait sprays . Results also indicate that type of protein used with phloxine B can dramatically influence its efficacy . Hydrolyzed proteins of corn origin, Mazoferm 802 and Nutriplus, and one from microbial origin, Coltec yeast broth, were best . Phloxine B-bait applications as complete coverage or alternate swaths reduced fly populations as well as 19.5 or 9.8% (AI) malathion-Captor 300 . Applications of phloxine B bait at concentrations of 0.12% phloxine B reduced populations as well as those applied at 0.48% (AI) . The fruit fly parasitoid Diachasmimorpha longicaudata was adversely affected when exposed to phloxine B-Nutriplus bait but not when exposed to the other proteins.

J Antibiot (Tokyo), 2001 Oct, 54(10), 783 - 8
A facile and effective screening method for p21WAF1 promoter activators from microbial metabolites; Nie L et al.; We have developed a novel p21WAF1 promoter activator screening system based on rapid and facile luciferase activity assay of a model cell system (H1299/tsp53-luc cells), a stable luciferase expression cell line established by transfecting H1299/tsp53 cells with a reporter gene construct pWWP-Luc-BSD . This plasmid was constructed by subcloning the 2.4 kb p21WAF1 promoter and a 2.6 kb of luciferase cDNA fragment activated by the p21WAF1 promoter into a pMAM2-BSD expression vector containing the blasticidin S deaminase gene (BSD) . A BSD-resistant clone H1299/tsp53-luc#4, showing the highest response to p53 activation (by temperature shift from 37 degrees C to 32 degrees C) by luciferase production, was used for screening microbial culture broths . Among approximately 1200 screened samples, trichostatin A related compounds and a new compound, lucilactaene, were isolated . This provides an effective and facile screening system for p21WAF1 promoter activators which should be of considerable value in the rapid identification of new anticancer agents.

Infect Control Hosp Epidemiol, 2001 Oct, 22(10), 635 - 9
An in-use evaluation of an alcohol-based pre-surgical hand disinfectant; Bryce EA et al.; OBJECTIVE: To determine whether alcohol hand disinfection is an effective alternative to traditional agents for the pre-surgical scrub . DESIGN: A prospective clinical trial of a 70% isopropanol pre-surgical hand disinfectant . SETTING: The operating room suites at two hospital sites in British Columbia . METHODS: Cases were selected to evaluate both short and longer procedures . The hand disinfectant was compared to agents in current use as surgical scrubs (4% chlorhexidine and 7.5% povidone-iodine) . Surgical technique and glove use were not modified . Pre- and postoperative fingertip impression and "glove-juice" cultures were used to determine microbial burden, and hands were evaluated for skin integrity . RESULTS: There was no statistical difference between the microbial hand counts following use of the alcohol-based product or the current agents, for cases less than 2 hours' duration . Comparison of longer surgical cases revealed significantly better pre- and postoperative culture results with the alcohol hand rinse, but analysis of matched pairs showed no significant difference in microbial counts . The alcohol hand rinse was equivalent to the operative scrub in terms of skin integrity and user acceptability . CONCLUSION: An alcohol hand rinse was equivalently effective in reducing microbial hand counts as the traditional pre-surgical scrub, both immediately after hand disinfection and at the end of the surgical procedure.

Allergy Asthma Proc, 2001 Nov-Dec, 22(6), 337 - 40
Endotoxin: friend or foe?
Liu AH, Redmon AH Jr.
Endotoxin in our living environment has been of increasing interest to our global community of allergists . Initially and largely studied for its capacity to mediate septic shock (and earning its "-toxin" suffix), we have since come to understand that endotoxin in the dust of many occupational settings also is an occult respiratory culprit, inducing airflow obstruction and aggravating asthma and allergies . More recently, environmental endotoxin has been implicated as a microbial exposure in early childhood that may have an atopy-protective effect by augmenting early Th1-type immune development . Although seemingly paradoxical, endotoxin's dual nature ultimately may serve to enlighten our understanding of how such bioactive exposures can interact with and guide our immune systems in both health and disease.

World J Surg, 2001 Dec, 25(12), 1512 - 8
Changes in bacterial concentration in the liver correlate with that in the hepaticojejunostomy after bile duct reconstruction: implication in the pathogenesis of postoperative cholangitis; Chuang JH et al.; Postoperative cholangitis is a frequent and unpredictable complication of unknown etiology following bile duct reconstruction (BDR), particularly for biliary atresia . This study was undertaken to correlate the growth of bacteria in the hepaticojejunostomy with that in the liver after BDR . Quantitative bacterial culture was done on the specimens taken from the liver and from the hepaticojejunostomy at 1 week (group 1, n = 7), 1 month (group 2, n = 7), and 2 months (group 3, n = 7) following BDR with Roux-en-Y hepaticojejunostomy in piglets after 2 weeks of common bile duct ligation . The histological examination of the liver and the hepaticojejunostomy, as well as serial monitoring of hemogram and liver function tests, were performed to correlate the findings with the bacterial concentration of the liver and the hepaticojejunostomy following BDR . The bacterial concentration of the hepaticojejunostomy, expressed as log10 colony-forming units per gram (log10 CFU/g) of the hepaticojejunostomy, showed a progressive decrease from 8.38 +/- 1.36 in group 1, 7.07 +/- 2.54 in group 2, to 3.56 +/- 1.31 in group 3 (p = 0.001) . The log10 CFU/g of the liver also showed a progressive decrease from 5.02 +/- 1.59 in group 1, 3.16 +/- 1.56 in group 2, to 2.19 +/- 1.09 in group 3 (p = 0.006) . There was a significant positive correlation of the log10 CFU/g of the liver (n = 21) with that of the hepaticojejunostomy (n = 21) following BDR (r = 0.600, p = 0.004) . Most of the infectious pathogens isolated from the liver were also isolated from the hepaticojejunostomy . The changes in hemoglobin, bilirubin, albumin, and ammonia significantly correlated with the changes of the bacterial concentration of the liver . The results of the study suggests that hepatic bacterial proliferation after BDR is significantly affected by microbial overgrowth in the bilioenteric anastomosis and is associated with deteriorated liver function and hemogram.

Environ Sci Technol, 2001 Dec 15, 35(24), 4783 - 8
Hydrogen concentrations in sulfate-reducing estuarine sediments during PCE dehalogenation; Mazur CS et al.; Despite recent progress made evaluating the role of hydrogen (H2) as a key electron donor in the anaerobic remediation of chloroethenes, few studies have focused on the evaluation of hydrogen thresholds relative to reductive dehalogenation in sulfidogenic environments . Competition for hydrogen exists among microbial populations in anaerobic sediments, and direct evidence indicates that lower hydrogen thresholds are observed with more energetically favorable electron-accepting processes . This study examined aqueous hydrogen concentrations associated with sulfate reduction and perchloroethylene (PCE) dehalogenation in anoxic estuarine sediment slurry microcosms and evaluated the competition for H2-reducing equivalents within these systems . After an initial lag period of 13 days, PCE was reductively transformed to trichloroethylene (TCE) . During the time of continuous PCE dehalogenation, a significantly (P < 0.05) lower hydrogen concentration (0.5 nM) was observed in the sediment slurries amended with PCE as compared to slurries without PCE (0.8 nM) . Sulfate reduction to sulfide was observed in all sediment slurries, but in microcosms actively dechlorinating PCE, the amount of reducing equivalents directed to sulfate reduction was approximately half the amount in sediment slurries without PCE . These findings provide evidence that a lower hydrogen threshold exists in anoxic estuarine sediment slurries with PCE as a terminal electron acceptor as compared to sediment slurries in which sulfate reduction was the predominant electron-accepting process . Furthermore, our results utilizing the inhibitor molybdate indicated that H2-utilizing methanogens may have the potential to effectively compete with dechlorinators for hydrogen when sulfate reduction is initially inhibited.

Environ Sci Technol, 2001 Dec 15, 35(24), 4719 - 25
Processes of nickel and cobalt uptake by a manganese oxide forming sediment in Pinal Creek, Globe mining district, Arizona; Kay JT et al.; A series of column experiments was conducted using manganese oxide coated sediments collected from the hyporheic zone in Pinal Creek (AZ), a metal-contaminated stream, to study the uptake and retention of Mn, Ni, and Co . Experimental variables included the absence (abiotic) and presence (biotic) of active Mn-oxidizing bacteria, the absence and presence of dissolved Mn, and sediment manganese oxide content . Uptake of Mn under biotic conditions was between 8 and 39% higher than under abiotic conditions . Continuous uptake of Mn due to biotic oxidation was evident from extraction of column sediments . Manganese uptake is hypothesized to initially occur as adsorption, which led to subsequent surface and/or microbial oxidation . Complete breakthrough of Ni within 100 pore volumes indicated no process of continuous uptake and was modeled as an equilibrium adsorption process . Nickel uptake in the presence of dissolved Mn was 67-100% reversible . Sediment extractions suggest that Ni uptake occurred through weak and strong adsorption . Continuous uptake of cobalt increased with sediment manganese oxide content, and Co uptake was up to 75% greater under biotic than abiotic conditions . Cobalt uptake was controlled by both existing and newly formed manganese oxides . Only a small amount of Co uptake was reversible (10-25%) . XANES spectral analysis indicated that most Co(II) was oxidized to Co(III) and probably incorporated structurally into manganese oxides . Although manganese oxides were the primary phase controlling uptake and retention of Mn, Ni, and Co, the mechanisms varied among the metals.

Appl Environ Microbiol, 2002 Jan, 68(1), 289 - 96
Duplicate copies of genes encoding methanesulfonate monooxygenase in Marinosulfonomonas methylotropha strain TR3 and detection of methanesulfonate utilizers in the environment; Baxter NJ et al.; Marinosulfonomonas methylotropha strain TR3 is a marine methylotroph that uses methanesulfonic acid (MSA) as a sole carbon and energy source . The genes from M . methylotropha strain TR3 encoding methanesulfonate monooxygenase, the enzyme responsible for the initial oxidation of MSA to formaldehyde and sulfite, were cloned and sequenced . They were located on two gene clusters on the chromosome of this bacterium . A 5.0-kbp HindIII fragment contained msmA, msmB, and msmC, encoding the large and small subunits of the hydroxylase component and the ferredoxin component, respectively, of the methanesulfonate monooxygenase, while a 6.5-kbp HindIII fragment contained duplicate copies of msmA and msmB, as well as msmD, encoding the reductase component of methanesulfonate . Both sets of msmA and msmB genes were virtually identical, and the derived msmA and msmB sequences of M . methylotropha strain TR3, compared with the corresponding hydroxylase from the terrestrial MSA utilizer Methylosulfonomonas methylovora strain M2 were found to be 82 and 69% identical . The msmA gene was investigated as a functional gene probe for detection of MSA-utilizing bacteria . PCR primers spanning a region of msmA which encoded a unique Rieske {2Fe-2S} binding region were designed . These primers were used to amplify the corresponding msmA genes from newly isolated Hyphomicrobium, Methylobacterium, and Pedomicrobium species that utilized MSA, from MSA enrichment cultures, and from DNA samples extracted directly from the environment . The high degree of identity of these msmA gene fragments, compared to msmA sequences from extant MSA utilizers, indicated the effectiveness of these PCR primers in molecular microbial ecology.

J Food Prot, 2001 Dec, 64(12), 2020 - 6
Consumer acceptance of irradiated meat and poultry in the United States; Frenzen PD et al.; Food manufacturers in the United States are currently allowed to irradiate raw meat and poultry to control microbial pathogens and began marketing irradiated beef products in mid-2000 . Consumers can reduce their risk of foodborne illness by substituting irradiated meat and poultry for nonirradiated products, particularly if they are more susceptible to foodborne illness . The objective of this study was to identify the individual characteristics associated with willingness to buy irradiated meat and poultry, with a focus on five risk factors for foodborne illness: unsafe food handling and consumption behavior, young and old age, and compromised immune status . A logistic regression model of willingness to buy irradiated meat or poultry was estimated using data from the 1998-1999 FoodNet Population Survey, a single-stage random-digit dialing telephone survey conducted in seven sites covering 11% of the U.S . population . Nearly one-half (49.8%) of the 10,780 adult respondents were willing to buy irradiated meat or poultry . After adjusting for other factors, consumer acceptance of these products was associated with male gender, greater education, higher household income, food irradiation knowledge, household exposure to raw meat and poultry, consumption of animal flesh, and geographic location . However, there was no difference in consumer acceptance by any of the foodborne illness risk factors . It is unclear why persons at increased risk of foodborne illness were not more willing to buy irradiated products, which could reduce the hazards they faced from handling or undercooking raw meat or poultry contaminated by microbial pathogens.

J Food Prot, 2001 Dec, 64(12), 1988 - 95
Alfalfa seed germination and yield ratio and alfalfa sprout microbial keeping quality following irradiation of seeds and sprouts; Rajkowski KT et al.; Foods can be treated with gamma radiation, a nonthermal food process, to inactivate foodborne pathogens and fungi, to kill insects on or in fruits and vegetables, and to increase shelf life . Gamma irradiation is especially well suited for these treatments because of its ability to penetrate commercial pallets of foods . Irradiated fruits, vegetables, poultry, and hamburger have been received favorably by the public and are now available in supermarkets . The use of irradiation on fresh alfalfa sprouts was studied to determine its effect on keeping quality as related to aerobic microbial load . After an irradiation dose of 2 kGy, the total aerobic count decreased from 10(5-8) to 10(3-5) CFU/g, and the total coliform counts decreased from 10(5-8) to 10(3-0) CFU/g . The results showed that the sprouts maintained their structure after irradiation, and the keeping quality was extended to 21 days, which is an increase of 10 days from the usual shelf life . The effect of various doses of irradiation on alfalfa seeds as measured by percent germination and yield ratio (wt/wt) of sprouts was determined . There was little effect on the percent germination, but as the dose increased, the yield ratio of alfalfa sprouts decreased . As the length of growing time increased, so did the yield ratio of the lower dose irradiated seeds (1 to 2 kGy) . The irradiation process can be used to increase the shelf life of alfalfa sprouts, and irradiating alfalfa seeds at doses up to 2 kGy does not unacceptably decrease the yield ratio for production of alfalfa sprouts.

Ann Chim, 2001 Sep-Oct, 91(9-10), 577 - 85
Organic content of particulate matter in the atmosphere of Ouargla City, Algeria; Yassaa N et al.; The particle-bound organic compounds comprising n-alkanes, n-alkanoic monocarboxylic acids, polynuclear aromatic hydrocarbons (PAH) and nitrated polynuclear aromatic hydrocarbons (NPAH) were investigated in ambient air of the Ouargla city area (Algeria) during a short campaign performed in November 1999 . The distribution profile of n-alkanes was consistent with the petrogenic origin of aerosols, suggesting that they were related to torching processes of crude oil refuses in the petroleum extraction field located not far from Ouargla . Instead, the presence of n-alkanoic acids of low molecular weight was indicative of microbial activity experienced by the site . PAH levels were low when compared to other polluted areas . The presence of 2-nitrofluoranthene and 2-nitropyrene, which can result from in-situ nitration of fluoranthene and pyrene, was concurrent with the substantial depletion of the most reactive among PAH, suggesting that photochemical processes influence the composition of the Saharian atmosphere.

Parasitology, 2001, 123 Suppl, S159 - 67
Mechanisms of molluscan host resistance and of parasite strategies for survival; Bayne CJ et al.; In parallel with massive research efforts in human schistosomiasis over the past 30 years, persistent efforts have been made to understand the basis for compatibility and incompatibility in molluscan schistosomiasis . Snail plasma contains molecules that are toxic to trematodes, but these seem to kill only species that never parasitize the mollusc used as the source of plasma . A sporocyst will be killed actively by haemocytes alone if they are from a snail that is resistant to the trematode . Oxygen-dependent killing mechanisms play a major role . Enzymes such as NADPH oxidase, superoxide dismutase, myeloperoxidase and nitric oxide synthase are critical components of the putative killing pathways . Metabolic intermediates such as hydrogen peroxide and nitric oxide appear to be more important against trematodes than the shorter-lived intermediates that are more important in anti-microbial defences . Products secreted by trematode larvae influence the physiology of snail haemocytes, implying active counter-defences mounted by the parasite, but these remain largely unexplored . A possible molecular basis for the susceptibility/resistance dichotomy in molluscan schistosomiasis is suggested to be deficient forms of enzymes in the respiratory burst pathway, and a selective disadvantage for schistosome resistance is an integral component of this model.

Huan Jing Ke Xue, 2001 Sep, 22(5), 7 - 11
{N2O emission by trees under natural condition}; Xu H et al.; The sources of N2O, one of the major greenhouse gases in the atmosphere, are still unclear till now . It was previously thought that only soil microbial process produced N2O . The aim of this study was to provide evidences that plant can emit N2O under the natural condition . The measurement of aboveground vertical profiles of N2O concentrations were conducted within coniferous-deciduous mixed forests in Changbai Mountain, China . Significant high N2O concentrations were found at or near the height of canopy . High concentrations in profiles were 3.03% to 64.9% higher than the "normal concentrations" . Differences between the high concentrations and the "normal concentrations" were statistically significant . The simultaneous occurrence of high concentrations at/nearby the canopy height and normal concentrations at the trunk space height indicated an efflux of N2O from foliage to atmosphere . This study afforded evidences supporting that plant per se, besides forest soil, was an important source of atmospheric N2O in a forest ecosystem.

Huan Jing Ke Xue, 2001 Sep, 22(5), 122 - 5
{Composting process of municipal solid waste with high effective complex microbial community}; Xi B et al.; The effects of high effective complex microbial community (MECMC) in a combination composting process of municipal solid waste (MSW) and sludge were examined through inspecting biomass, temperature, organic matter, the carbon, nitrogen ration (C/N) . Composting was performed under such conditions: The ratio of MSW:sewage:sawdust:composting of 45:25:15:15 (W/W) and organic matter of 60%, total N of 1.4%, total P of 0.69%, total K of 1.25%, original moisture of 58.5%, original ratio of C/N of 30, aration flow of 0.8 L/min.kg(volatile organic matter) . The rates of adding HECMC to treatment 1, treatment 2 and treatment 3 were 2%, 3%, 5% respectively . Control experiment added 3% inactive HECMC . The day of MSW and sludge mature in control experiment, treatment 1, treatment 2 and treatment 3 were 30, 24, 18 and 12, respectivly . This indicated HECMC were effective to compose organic matter and speedup composting change into humus . There are many active bacteria in composting which are favorable for soil.

J Bioenerg Biomembr, 2000 Jun, 32(3), 259 - 67
Metabolic functions of microbial nucleoside diphosphate kinases; Bernard MA et al.; This article summarizes research from our laboratory on two aspects of the biochemistry of nucleoside diphosphate kinase from Escherichia coli--first, its interactions with several T4 bacteriophage-coded enzymes, as part of a multienzyme complex for deoxyribonucleoside triphosphate biosynthesis . We identify some of the specific interactions and discuss whether the complex is linked physically or functionally with the T4 DNA replication machinery, or replisome . Second, we discuss phenotypes of an E . coli mutant strain carrying a targeted deletion of ndk, the structural gene for nucleoside diphosphate kinase . How do bacteria lacking this essential housekeeping enzyme synthesize nucleoside triphosphates? In view of the specific interactions of nucleoside diphosphate kinase with T4 enzymes of DNA metabolism, how does T4 multiply after infection of this host? Finally, the ndk disruption strain has highly biased nucleoside triphosphate pools, including elevations of the CTP and dCTP pools of 7- and 23-fold, respectively . Accompanied by these biased nucleotide pools is a strong mutator phenotype . What is the biochemical basis for the pool abnormalities and what are the mutagenic mechanisms? We conclude with brief references to related work in other laboratories.

J Anim Sci, 2001 Nov, 79(11), 2908 - 12
Effect of feeding frequency on bacterial and fungal concentrations, pH, and other parameters in the rumen; Dehority BA et al.; Three sheep were fed a pelleted high-roughage diet either once, 6, or 24 times per day in a 3 x 3 Latin square trial . During each 21-d period, 14 d were allowed for adaptation followed by a 7-d collection period, in which samples for microbial counts were taken on d 1 and 5 and several rumen parameters were measured on d 2 and 6 . Bacterial concentrations were not different between feeding frequencies on the first sampling day but were higher (P < 0.05) on the second sampling day when the sheep were fed 24 times a day . Fungal concentrations were not different among feeding frequencies on either sampling day . No effects of feeding frequency were observed for the concentration of cellulolytic bacteria or fungi . On d 2, ruminal volume was larger (P < 0.05) with six feedings than with one feeding and fluid volume turnover was greater (P < 0.05) when sheep were fed 24 times per day . Rumen pH values were higher (P < 0.01) on both d 1 and 5 when the sheep were fed once a day and the percentage of rumen dry matter was highest (P < 0.02) with 24 feedings . These findings would suggest that if the same amount of a given diet is fed daily, the number of feedings does not markedly affect microbial concentrations, rumen volume, or liquid turnover time.

PDA J Pharm Sci Technol, 2001 Nov-Dec, 55(6), 393 - 416
Method for qualifying microbial removal performance of 0.1 micron rated filters . Part III: bacterial challenge tests on 0.2/0.22 and 0.1 micron rated filter cartridges with Hydrogenophaga (formerly Pseudomonas) pseudoflava; Sundaram S et al.; We have previously reported on the preliminary characterization of Hydrogenophaga (formerly Pseudomonas) pseudoflava for potential use as a standard challenge organism to qualify 0.1 microm rated filters . This article reports on the retention efficiencies of a large panel of 0.2/0.22 microm and 0.1 microm rated filter cartridges for H . pseudoflava (ATCC 700892) versus the retention capabilities of the same filters for Brevundimonas diminuta (ATCC 19146) . A total of thirty-two 0.2/0.22 microm rated filter cartridges, spanning nine different "sterilizing grade" filter types from four different filter manufacturers, were challenged with H . pseudoflava at challenge levels exceeding 10(7) cfu/cm2 . H . pseudoflava was shown to penetrate every 0.2/0.22 microm rated filter tested, with log titer reduction (LTR) values ranging from 3.5 to 7.7 logs . H . pseudoflava was shown to be more penetrative than B . diminuta under the same challenge conditions . B . diminuta was fully retained by nineteen of the twenty 0.2/0.22 microm rated filters that were challenged with both organisms . In the case of 0.1 microm rated filters, eighteen filter cartridges, spanning five different filter types from three manufacturers were tested . H . pseudoflava was consistently retained by four out of the five filter types tested, with LTR values in excess of 11.5 to 12.2 logs . The 0.1 microm rated filter type that was penetrated by H . pseudoflava has been previously demonstrated to be not fully retentive for naturally occurring bacteria . The data show that H . pseudoflava penetrates 0.2/0.22 microm rated filters just as readily as B . diminuta penetrates 0.45 microm rated filters . In addition, titer reductions provided by 0.2/0.22 microm rated filters for H . pseudoflava are comparable to those reported for A . laidlawii mycoplasma, albeit under different conditions . This study demonstrates that H . pseudoflava meets all criteria for use as a standard organism for qualifying the microbial removal performance of 0.1 microm rated filters for enhanced sterility assurance.

PDA J Pharm Sci Technol, 2001 Nov-Dec, 55(6), 373 - 92
Method for qualifying microbial removal performance of 0.1 micron rated filters . Part II: preliminary characterization of Hydrogenophaga (formerly Pseudomonas) pseudoflava for use as a standard challenge organism to qualify 0.1 micron rated filters; Sundaram S et al.; In this article, we report on the preliminary characterization of Hydrogenophaga (formerly Pseudomonas) pseudoflava for potential use as a standard challenge organism to qualify 0.1 microm rated filters . Filter-cloned H . pseudoflava (ATCC 700892) was easily cultured in a low nutrient broth (R2A broth) under standard laboratory conditions, reaching high titers of 10(8)-10(9) cfu/mL within 48-65 hours of incubation at 25+/-5 degrees C . Under these conditions, H . pseudoflava is a rod-shaped bacterium, averaging 0.25+/-0.03 microm by 1.65+/-0.35 microm, and appears to be smaller than Brevundimonas diminuta in width (0.31+/-0.03 microm), but somewhat longer in length (0.88+/-0.19 microm), which may partly explain the observed penetration . In total, thirty-five 0.2/0.22 microm rated filter discs, spanning five different "sterilizing grade" filter types from two different filter manufacturers were challenged with H . pseudoflava . In all cases, H . pseudoflava was shown to consistently penetrate every 0.2/0.22 microm rated filter disc tested . These tests also spanned three different challenge durations, including short-term challenges (30-40 minutes), and two different challenge fluids . The use of serial (double) 0.22 mm rated filters, which is a common industry practice to reduce the prefiltration bioburden to the final "sterilizing" filter, was also shown to be inadequate to fully retain H . pseudoflava under the challenge condition used . In contrast, two different 0.1 microm rated filter types functionally qualified with a specified high titer reduction claim for Acholeplasma laidlawii, were shown to consistently and fully retain H . pseudoflava, and retention by these two filter types was shown to be robust and independent of the challenge duration.

PDA J Pharm Sci Technol, 2001 Nov-Dec, 55(6), 346 - 72
Method for qualifying microbial removal performance of 0.1 micron rated filters . Part I: characterization of water isolates for potential use as standard challenge organisms to qualify 0.1 micron rated filters; Sundaram S et al.; Although 0.1 microm rated filters intended for pharmaceutical sterilization applications have been commercially available for at least 15 years, there is no industry-wide standard for qualifying the microbial removal performance of these filters . In this article, we report on the bacterial challenge methodology used to screen four bacterial species for potential utility as a standard challenge organism to qualify 0.1 microm rated filters . These isolates were, in their natural state, demonstrated to penetrate 0.2/0.22 microm rated filters in prior studies . In the screening challenges described in this study, three out of these four candidates tested demonstrated consistent penetration of one 0.22 microm rated filter type tested (when cultured in a low nutrient medium under standard laboratory conditions) . These included 6204-22 (FAME ID Acidovorax avenae citrulli), 6266-15 (FAME ID Comamonas acidovorans), and 6266-34 (FAME ID Hydrogenophaga pseudoflava) . Of these, H . pseudoflava (6266-34) was chosen for additional experiments with other 0.2 microm rated filter membranes . In total, seventeen 0.2 and 0.22 microm rated filter discs, spanning five different "sterilizing grade" filter types from three different filter manufacturers were tested . H . pseudoflava penetration was observed for every filter tested . Under the same challenge conditions, H . pseudoflava was consistently retained by a 0.1 microm rated hydrophilic PVDF (polyvinylidenefluoride) filter with a specified high titer reduction claim for Acholeplasma laidlawii . In order to ensure selection of the most stable penetrative phenotype (i.e., select for nonrevertants), H . pseudoflava was subjected to three rounds of "filter cloning," and these results are described herein . The advantages of using H . pseudoflava for qualifying the microbial removal performance of 0.1 microm rated filters are also discussed.

Xenobiotica, 2001 Nov, 31(11), 799 - 809
Oxidation of phenothiazine and phenoxazine by Cunninghamella elegans; Sutherland JB et al.; 1 . To determine the ability of fungi to metabolize sulphur- and oxygen-containing azaarenes, Cunninghamella elegans ATCC 9245 was grown in 125-ml flasks containing fluid Sabouraud medium . The cultures and controls were incubated at 28 degrees C with shaking and dosed with 16.7 mM phenothiazine or phenoxazine . After incubation for 72h, the mycelia and filtrates were extracted with ethyl acetate and the combined residues analysed by high-performance liquid chromatography . Residual phenothiazine and phenoxazine were 21 and 22%, respectively, of the total UV absorbance at 254 nm . 2 . The metabolites were identified by mass spectrometry and proton nuclear magnetic resonance spectroscopy . The fungus oxidized phenothiazine to phenothiazine sulphoxide, 3-hydroxyphenothiazine sulphoxide, phenothiazin-3-one, and 3-hydroxyphenothiazine and oxidized phenoxazine to phenoxazin-3-one . 3 . Three of the four compounds produced by C . elegans from phenothiazine were identical to those produced by mammals, supporting the use of the fungus as a microbial model for drug metabolism.

Curr Opin Pharmacol, 2001 Oct, 1(5), 513 - 20
Proteomics: an holistic analysis of nature's proteins; Hebestreit HF; Proteomics has matured to a technology platform way beyond two-dimensional gel electrophoresis, delivering on its promise to identify structure, function and cellular localization of all proteins expressed in a cell at a given time . Major achievements in the past year include mapping the proteome of human and microbial cells, improvements in two-dimensional gel electrophoresis and mass spectrometric analysis, and the development of protein arrays and biochips.

Environ Toxicol Chem, 2001 Dec, 20(12), 2717 - 24
Fate of atrazine and alachlor in redox-treated ferruginous smectite; Xu JC et al.; The oxidation state of structural iron (Fe) in clay minerals exerts a large influence on clay surface chemistry and may affect the adsorption and degradation of pesticides in the environment . This effect, however, has been little investigated . In the present study, herbicides atrazine and alachlor were reacted with ferruginous smectite (sample SWa-1) in its oxidized, reduced (either chemically or bacterially), and reduced-reoxidized states . In some experiments the herbicide was labeled with 14C . Gas chromatography/mass spectrometry (GC-MS) was also used to detect alachlor degradation products . Compared to oxidized clays, reduction by both chemical and microbial treatments decreased the concentration of both herbicides in the surrounding solution . Reoxidized clay exhibited behavior similar to the oxidized clay . Hydrolysis-dechlorination of atrazine occurred in the presence of chemically reduced SWa-1, and GC-MS analysis of alachlor revealed at least 14 degradation products after treatment with reduced clay and only two with the oxidized clay . Interaction of atrazine and alachlor with the clay may be through a H bond with the waters of hydration surrounding interlayer cations, the extent of which should increase with increasing acidity; but under reduced conditions, the validity of this model is unclear . Reduction of structural Fe may affect pH-dependent phenomena in two ways: The increased surface charge density increases the number of hydrated interlayer cations, thereby enhancing surface acidity, and increased electron density at basal surface oxygens increases their Bronsted basicity . Atrazine could, therefore, adsorb and/or degrade through either acid or alkaline hydrolysis pathways . Increased reduction potential of the reduced clay surfaces may also promote degradation.

Mar Pollut Bull, 2001 Nov, 42(11), 1082 - 6
The effects of freshwater flushing on marine heterotrophic protists--implications for ballast water management; Hulsmann N et al.; Survivorship of ballast-entrained marine heterotrophic protists was examined following freshwater flushing . The recovered taxa, including typical marine rhizopods such as Platyamoeba murchelanoi, Labyrinthula spp, Pontifex maximus, Thecamoeba orbis, and the ciliate Condylostoma arenarium, were reared in waters of various salinities . After 2 months, the original salinity subsample retained five protist taxa, the freshwater six, including the amoeba Cochliopodium bilimbosum, the brackish water 22 taxa, and the seawater 19 taxa . Since protists form a major component of marine microbial food webs, their survival may be instrumental in supporting complex ballast-entrained food webs . Our study raises questions as to the reliability of open-ocean exchange (OOE) or freshwater flushing as effective control measures.

Arthritis Rheum, 2001 Dec, 44(12), 2879 - 87
TIA-1 regulates the production of tumor necrosis factor alpha in macrophages, but not in lymphocytes; Saito K et al.; OBJECTIVE: To determine whether TIA-1 differentially regulates the production of tumor necrosis factor a (TNFalphalpha) in macrophages and lymphocytes . METHODS: Peritoneal macrophages derived from wild-type and TIA-1-/- mice were cultured in the absence or presence of lipopolysaccharide (LPS) before comparison of the production of TNFalpha protein by intracellular flow cytometry and the secretion of TNFalpha protein by enzyme-linked immunosorbent assay . In parallel experiments, splenocytes were cultured in the absence or presence of concanavalin A (Con A), phorbol myristate acetate (PMA)/ionomycin, or anti-CD3/anti-CD28 before comparing the production of TNFalpha protein . Finally, the relative expression of TIA-1 protein in macrophages and splenocytes was compared using immunoblotting analysis . RESULTS: LPS-activated peritoneal macrophages derived from TIA-1-/- mice produced significantly more TNFalpha than macrophages from wild-type controls . In contrast, splenic lymphocytes (CD3+, CD4+, or CD8+) derived from wild-type and TIA-1-/- mice produced similar amounts of TNFalpha in response to Con A, PMA/ionomycin, or anti-CD3/anti-CD28 . Lymphocytes and macrophages expressed similar amounts of TIA-1 protein, indicating that differential expression of TIA-1 cannot account for these results . CONCLUSION: TIA-1 is the target of a regulatory pathway that operates in activated macrophages, but not in activated lymphocytes . Developing drugs that target this pathway might prevent the pathologic overexpression of TNFalpha without subverting the T lymphocyte response to microbial pathogens.

Bioresour Technol, 2002 Jan, 81(2), 163 - 5
Bioconversion of eucalyptus bark waste into soil conditioner; Yadav KR et al.; An optimized protocol for the bioconversion of eucalyptus bark was devised . It comprised: (i) mechanical reduction in bark size to 0.5-3.0 cm, (ii) moistening to 60-65%, (iii) fortification with ligninase-rich fungus Volvariella sp . (S-1) and 2% urea and (iv) maintenance of this composting mix under aerobic and ambient condition for 14-15 weeks . The resulting bark soil conditioner (BSC) was an easily crumbling, reddish brown biomass, with physico-chemical and microbial properties which would enrich soil fertility/productivity.

Bioresour Technol, 2002 Jan, 81(2), 155 - 7
Decolourisation of the textile dye Astrazon Red FBL by Funalia trogii pellets; Yesilada O et al.; The effects of various conditions such as initial pH, dye concentrations, amount of pellet, temperature and agitation on decolourising activity of Funalia trogii were investigated . These, except initial pH, were all found to be important for dye decolourising activity of F . trogii . The decolourisation of the dye involved adsorption of the dye compound by fungal pellets at the initial stage, followed by the decolourisation through microbial metabolism . Heat-killed pellets were also tested for their ability to decolourise Astrazon Red dye . These pellets adsorbed the dye and 55% decolourisation was obtained in 24 h . But at the second cycle there was only 24% decolourisation . Our observation showed that Astrazon Red dye decolourisation by heat-killed pellets was mainly due to biosorption . The longevity of the decolourisation activity of F . trogii pellets was also investigated in repeated batch mode . Variations in the amount of pellet increased % decolourisation and stability of pellets.

Indoor Air, 2001 Dec, 11(4), 223 - 31
Sign of eye irritation in female hospital workers and the indoor environment; Smedbold HT et al.; The relationships between indoor environment factors and signs of eye irritation were studied in a sample of female personnel with (n = 112) and without (n = 64) mucous membrane symptoms working in 36 nursing departments at geriatric hospitals in the municipality of Trondheim, Norway . The indoor climate was characterised by high room temperature (interquartile range 23.0-23.7 degrees C), low relative air humidity (interquartile range 17-26%) and high outdoor airflow rate, as indicated by low indoor levels of carbon dioxide (interquartile range 490-650 ppm) . An altered microbial flora was observed in nine of the departments, and Aspergillus fumigatus was found in three of these . Five of the hospitals were situated in urban parts of the municipality near roads with heavy traffic . In the initial analyses decreased tear-film stability (BUT) was observed in subjects working in departments with increased dust settlement rate (P = 0.03), in hospitals situated in urban areas with heavy traffic (P < 0.001), and in subjects working in departments with presence of A . fumigatus (P = 0.04) . Increased conjunctival staining (CS) was related to alteration of the microbial flora . In the final multivariate analyses statistically significant relationships were observed between BUT and dust settlement rate and urban vicinity of workplace and between CS and alteration of the microbial flora.

Isotopes Environ Health Stud, 2001, 37(2), 101 - 12
Origin and distribution of sulphate in surface waters of the Mansfeld mining district (Central Germany)--a sulphur isotope study; Strauch G et al.; In the Mansfeld region (Central Germany) copper mining contributed to an enormous pollution of the environment . Metal- and sulphate-bearing sediments and leachates emerge from the former copper smelters and mining waste heaps, spread along local rivers and finally reach the Saale river . A sulphur isotope study on water and stream sediments was performed along the River "Bose Sieben" and from its tributaries to determine the different sulphur sources . Four major sulphur sources exist in the area: metal sulphide mineralisations (Kupferschiefer), met alliferous sulphidic flue dust, slag, and anhydrite and gypsum of Permian and Triassic age . We obtained delta34S(SO4)-values in water samples varying from +4 per thousand to -18 per thousand CDT, clearly reflecting the input of sulphate from different sources . Sulphate from the oxidation of sulphidic mining residues is restricted to the mining area and cannot be traced for more than 5 km downstream . The major source for sulphate is the dissolution of gypsum and anhydrite . The sulphur isotope composition in dissolved and sedimentary adsorbed sulphate differs only slightly from each other . Microbial dissimilatory sulphate reduction can not be excluded in the shallow sediment layers.

Clin Exp Rheumatol, 2001 Sep-Oct, 19(5 Suppl 24), S6 - 12
Behçet's disease: an update on the pathogenesis; Gul A; Behcet's disease is a chronic inflammatory disorder of unknown etiology . It has long been postulated that immunological abnormalities, which are possibly induced by microbial pathogens in genetically susceptible individuals, are important in its pathogenesis . Recent findings have both supported the significance of genetic factors and better defined the nature of inflammation in Behcet's disease . Molecular genetic studies have strengthened the primary association of HLA-B51 with Behcet's disease . The exact pathogenic mechanism of the HLA-B51 molecule is still unknown, and its contribution to the overall genetic susceptibility to Behcet's disease is estimated to be less than 20% . Spontaneous and/or induced overexpression of pro-inflammatory cytokines (mainly Th1 type) from various cellular sources seems responsible for the enhanced inflammatory reaction in Behcet's disease, and it may be associated with the genetic susceptibility . An antigen-driven immune response superimposed on this primed-state and induced by heat shock proteins or other peptides from different strains of streptoccocci or other microbial agents has been suggested to trigger manifestations of Behcet's disease . Endothelial activation/injury and the resultant occlusive vasculopathy may also contribute to the tissue damage.

J Protein Chem, 2001 Aug, 20(6), 431 - 53
Application of the S-pyridylethylation reaction to the elucidation of the structures and functions of proteins; Friedman M; Cysteine (Cys) and cystine residues in proteins are unstable under conditions used for acid hydrolysis of peptide bonds . To overcome this problem, we proposed the use of the S-pyridylethylation reaction to stabilize Cys residues as pyridylethyl-cysteine (PEC) protein derivatives . This suggestion was based on our observation that two synthetic derivatives formed by pyridylethylation of the SH group of Cys with either 2-vinylpyridine (2-VP) or 4-vinylpyridine (4-VP), designated as S-beta-(2-pyridylethyl)-L-cysteine (2-PEC) and S-beta-(4-pyridylethyl)-L-cysteine (4-PEC), were stable under acid conditions used to hydrolyze proteins . This was also the case for protein-bound PEC groups . Since their discovery over 30 years ago, pyridylethylation reactions have been widely modified and automated for the analysis of many structurally different proteins at levels as low as 20 picomoles, to determine the primary structures of proteins and to define the influence of SH groups and disulfide bonds on the structures and functional, enzymatic, medical, nutritional, pharmacological, and toxic properties of proteins isolated from plant, microbial, marine, animal, and human sources . Pyridylethylation has been accepted as the best method for the modification of Cys residues in proteins for subsequent analysis and sequence determination . The reaction has also been proposed to measure D-Cys, homocysteine, glutathione, tryptophan, dehydroalanine, and furanthiol food flavors . This integrated overview of the diverse literature on these reactions emphasizes general concepts . It is intended to serve as a resource and guide for further progress based on the reported application of pyridylethylation reactions to more than 150 proteins.

Int J Hyg Environ Health, 2001 Nov, 204(2-3), 111 - 21
Detection of microbial volatile organic compounds (MVOCs) produced by moulds on various materials; Fiedler K et al.; Twelve fungal species were screened for microbial volatile organic compounds (MVOCs): Aspergillus fumigatus, A . versicolor, A . niger, A . ochraceus, Trichoderma harzianum, T . pseudokoningii, Penicillium brevicompactum, P . chrysogenum, P . claviforme, P . expansum, Fusarium solani and Mucor sp . More than 150 volatile substances derived from fungal cultures have been analysed by head-space solid-phase microextraction (HS-SPME) . Each species had a defined MVOC profile which may be subjected to considerable modification in response to external factors such as cultivation on different substrata . The cultivation on different substrata changes the number and concentration of MVOCs . Species-specific volatiles may serve as marker compounds for the selective detection of fungal species in indoor environments . Examination of MVOCs from indoor air samples may become an important method in indoor air hygiene for the detection of type and intensity of masked contamination by moulds.

Antonie Van Leeuwenhoek, 2001 Oct, 80(2), 185 - 91
Cometabolic biosynthesis of copolyesters consisting of 3-hydroxyvalerate and medium-chain-length 3-hydroxyalkanoates by Pseudomonas sp . DSY-82; Kang HO et al.; A newly isolated strain, designated as Pseudomonas sp . DSY-82, synthesized medium-chain-length polyhydroxyalkanoate (MCL-PHA) copolyesters when grown on alkanoates from hexanoate to undecanoate as the sole carbon source . When used alone, butyrate and valerate supported the growth of the isolate but not PHA production . However, unusual polyesters containing 3-hydroxyvalerate, as well as various MCL 3-hydroxyalkanoate monomeric units, were synthesized when valerate was cofed with either nonanoate or 10-undecenoate, suggesting the formation of monomer units from both substrates . Concentrations of 3-hydroxyvalerate, 3-hydroxyoctanoate, and 3-hydroxydecanoate in the PHAs produced were significantly elevated by the addition of valerate, indicating that the incorporation of these monomer units to PHA occurred primarily through cometabolism . The total amount of these monomer units in the PHAs reached up to 30% . The PHAs produced in this study were most likely random copolyesters as determined by differential scanning calorimetric analysis . This is the first case of microbial synthesis of copolyesters consisting of 3-hydroxyvalerate and MCL 3-hydroxyalkanoate monomer units through cometabolism.

J Hosp Infect, 2001 Aug, 48 Suppl A, S33 - 6
Hand antiseptics: rubs versus scrubs, alcoholic solutions versus alcoholic gels; Pietsch H; This report describes three different investigations undertaken to demonstrate the advantage of fluid alcoholic hand disinfectants . In the first study, the skin compatibility of Sterillium, a liquid alcoholic rub-in hand disinfectant was compared with that of Hibiscrub, a water-based handwashing antiseptic . Using various parameters such as image analysis of removed squames (D-squames), skin roughness or transepidermal water loss, Hibiscrub was found to be significantly inferior to Sterillium . Hibiscrub caused skin irritation in 15 volunteers who could not complete the test . In a second study, the microbicidal efficacy of Sterillium and Hibiscrub was tested in surgical hand disinfection . The microbial reduction by Sterillium was significantly greater than that of Hibiscrub, immediately after application as well as after the surgical procedure . In a third study, certain alcoholic gels were tested according to the EN 1500 'hygienic hand disinfection' . None of the gels tested passed the EN 1500 within 30s . However, Sterillium met the EN 1500 requirement within 30s . We conclude that Sterillium is superior to Hibiscrub in terms of skin tolerance and microbicidal efficacy in surgical hand disinfection . It is also superior to alcoholic gels.

J Hosp Infect, 2001 Aug, 48 Suppl A, S23 - 8
Hand hygiene--comparison of international recommendations; Wendt C; The value of hand hygiene for the prevention of cross-infection was first observed in the middle of the 19th century . Since then, which procedure is the most suitable for hand hygiene has been repeatedly discussed and several different guidelines and recommendations have been published . The aim of this review is to compare different recommendations for hand hygiene regarding technique and indication . Medline, the internet and a personal library were searched to obtain as many written recommendations as possible . In addition, a small questionnaire was sent by e-mail to 20 international colleagues . As a result, written recommendations from 10 countries could be compared . Recommended methods of hand hygiene include handwashing (washing hands with plain soap), hygienic handwash (washing hands with medicated soap) and hygienic hand-rub (use of antiseptic rubs) . In most countries handwashing and hygienic handwash are the methods of choice and only in central European countries is hygienic hand-rub the preferred technique . Situations in which performance of hand hygiene is recommended are comparable . However, no single indication is recommended in all guidelines . Hand hygiene is most often recommended before performing invasive procedures and after microbial contamination . Guidelines should be clear and easy to follow for them to become standard of care . Thus, guidelines are needed that do not leave to the health care worker a decision as to whether hand hygiene is indicated.

Ying Yong Sheng Tai Xue Bao, 2001 Aug, 12(4), 553 - 6
{Effect of different tillage methods on rice growth and soil ecology}; Li H et al.; Field experiments were conducted in double-cropping rice field in South China during 1998-1999 to study the effect of different tillage methods on rice growth and soil ecology . The results showed that with rice scattering planting, no-tillage method caused a reduction of rice tillering, effective panicle, and filled grains of rice . Grain yield under no-tillage was 13.40% lower than that under conventional tillage, and the economic benefits decreased by 10.9% . Soil analysis showed that in no-tillage fields, soil bulk density and hardness were increased, soil porosity and available P and K were decreased, the amount of actinomyces and fungi was reduced, while that of soil bacteria was increased, and the enzyme activity was promoted . Minimum tillage and conventional tillage had similar soil physical and chemical properties, soil microbial quantity and enzyme activity . Minimum tillage could produce a 2.1% higher grain yield than conventional tillage, and increase the economic benefits by 11.0%.

Oncology (Huntingt), 2001 Nov, 15(11 Suppl 9), 15 - 9
Antifungal prophylaxis in hematopoietic stem cell transplant recipients; Marr KA; Efforts at preventing and treating fungal infection in hematopoietic stem cell transplant (HSCT) recipients must take into account the types of infections likely to be encountered during the different risk periods in hosts with different underlying risks . Given the emergence of molds as prevalent pathogens and the long duration of risk in allogeneic HSCT recipients, optimal antifungal prophylaxis would consist of treatment that can be given over a prolonged period and that would provide both anti-Candida and anti-Aspergillus activity . Optimal empiric therapy would consist of a broad-spectrum agent in the absence of more sensitive and specific methods for microbial diagnosis . Fluconazole (Diflucan) is currently the standard prophylactic agent for candidiasis, although mold-active agents and alternative strategies for polyene administration are being investigated . The gold standardfor empiric therapy is currently a polyene antifungal, yet an increased appreciation for amphotericin B-resistant yeasts and molds, and less toxic mold-active alternatives, might lead to the use of other compounds in the future . The recent development of multiple alternatives emphasizes our need to establish treatment algorithms that consider both the likely pathogens and potential toxicities.

J Environ Sci Health B, 2001 Nov, 36(6), 729 - 39
Degradation of isoxaben in soils and an aqueous system; Camper ND et al.; The degradation of isoxaben {N-{3-(1-ethyl-1-methylpropyl)-5-isoxazolyl}-2,6-dimethoxybenzamide} was studied in soil and in an aqueous system . Soil studies were conducted in Erlenmeyer flasks (treated with 1 microg/g isoxaben) and mineralization studies in Biometer flasks (treated with 1 microg/g unlabeled and 14C-isoxaben) incubated at 23 C . Degradation in the aqueous system was performed in Erlenmeyer flasks under aerobic and anaerobic conditions incubated at 23 degrees C . Incubation mixtures were extracted at selected times and analyzed for isoxaben and degradation products by HPLC with product identification confirmed by GC-MS . After 8 weeks, 78% and 23% of the total isoxaben disappeared in nonsterile and sterile soils, respectively . After 12 weeks, approximately 1% of the labeled isoxaben was recovered as CO2 in the Biometer flask experiments; no volatile products were detected, and 5% and 33% of the total radioactivity was recovered from the nonsterile and sterile soils, respectively . In the aquatic system after 8 weeks, isoxaben had decreased from 1microg/g to 0.1 and 0.004 microg/g under aerobic and anaerobic conditions, respectively . Degradation products detected from the soil studies were 3-nitrophthalic acid and 4-methoxyphenol, and 3-nitrophthalic acid in the aqueous system studies . Microbial activity was considered to be a major factor in the degradation of isoxaben in this study.

Eur Respir J, 2001 Nov, 18(5), 872 - 81
Infection: friend or foe in the development of atopy and asthma? The epidemiological evidence; von Mutius E; There is increasing evidence from epidemiological studies to support the "hygiene hypothesis" with regard to the development of allergic diseases . This review will discuss the epidemiological findings relating to atopy and asthma, separately . The strongest arguments in favour of the "hygiene hypothesis" are the findings relating early life day care attendance to a significantly reduced risk of atopy and asthma, and the strong link between sibship size and the occurrence of atopy as confirmed by numerous studies . Furthermore, data relating serological immune responses to certain infections such as hepatitis A and Toxoplasma gondii suggest a role for such infections, or alternatively the lack of hygiene, to be operative in the inhibition of the development of allergic immune responses . The effects described seem to be stronger for atopy than for asthma . Furthermore, biological plausibility for such a hypothesis can be found from findings of recent in vitro studies and from animal experiments . Epidemiological studies attempting to identify single infections as being harmful or beneficial could fail, because it may be the total burden of microbial stimuli rather than a certain infection that directs the maturing immune system into a Th1-type T-helper cell polarized response . The timing of the exposure seems to be essential for promoting beneficial or harmful effects, and may provide a "window of opportunity".

Eur Respir J, 2001 Nov, 18(5), 784 - 9
The impact of meconium ileus on the clinical course of children with cystic fibrosis; Evans AK et al.; The present study was designed to compare the clinical course of children diagnosed with cystic fibrosis (CF) in infancy due to the presence of meconium ileus (MI) with children diagnosed by way of a newborn screening programme (non-MI) . A matched case-control study design was used . Matching was performed on the basis of sex and date of birth . All children born in New South Wales, Australia after 1980 and who had attended the CF clinic at The Children's Hospital at Westmead since diagnosis were included as possible cases or controls . Parameters pertaining to the clinical course were compared in 39 matched pairs . MI children had a significantly worse pulmonary status . The forced expiratory volume in one second was 16.3 +/- 5.2% higher (p<0.001, n=21 pairs) and the forced vital capacity value 10.5 +/- 4.7%, higher (p<0.05, n=21 pairs) in non-MI children . The difference between the pairs (18.6 +/- 4.4 MI and 20.5 +/- 3.4 non-MI) in the Shwachman chest radiograph score was statistically significant (p<0.05, n=39 pairs) . There were no significant differences in any other assessed parameters, such as height, weight, the presence of liver function abnormalities, the frequency of hospitalization or airway microbial colonization . Meconium ileus may be an early indication of a more severe phenotype of cystic fibrosis . This was suggested by the significantly lower pulmonary function found in children with a history of meconium ileus compared to age- and sex-matched children who did not have meconium ileus.

Eur J Med Chem, 2001 Nov-Dec, 36(11-12), 899 - 907
Synthesis and anti-microbial activities of some pyridinium salts with alkoxymethyl hydrophobic group; Pernak J et al.; A novel class of functionalized cationic surfactant has been obtained . The work-up procedure of synthesis is very simple, the yield is high and the pyridinium salts with alkoxymethyl hydrophobic group are easily purified . All the salts examined showed anti-microbial activities . Some of them exhibited strong activity and wide anti-bacterial spectra similar to the activity of benzalkonium chloride . The relationship between chemical structure and anti-microbial activity was analysed by the QSAR method.

Phytochemistry, 2002 Jan, 59(1), 39 - 44
Microbial metabolism of partheniol by Mucor circinelloides; Maatooq GT; Six new partheniol metabolites were isolated from the biotransformation reaction with Mucor circinelloides ATCC 15242 . These metabolites are: humula-1(10), 4, 7-trien-6alpha-ol 2, maali-3-en-8alpha-ol 3, aromadendrane-4alpha, 8alpha, 10alpha-triol 4, maaliane-4alpha, 8alpha, 9alpha-triol 5, maaliane-5alpha, 8alpha, 9alpha-triol 6, 5(9), 6-tricyclohumulane-4alpha, 8alpha, 10alpha-triol 7 . The structural assignments of these metabolites were made possible by different spectroscopic means.

Immunity, 2001 Dec, 15(6), 971 - 83
CD40 is a cellular receptor mediating mycobacterial heat shock protein 70 stimulation of CC-chemokines; Wang Y et al.; The 70 kDa mycobacterial heat shock protein (Mtb HSP70) stimulates mononuclear cells to release CC-chemokines . We now show that this function of Mtb HSP70, but not human HSP70, is dependent on the cell surface expression of CD40 . Deletion of the CD40 cytoplasmic tail abolished, and CD40 antibody inhibited, Mtb HSP70 stimulation of CC-chemokine release . Mtb HSP70 stimulated THP1, KG1 cells, and monocyte-derived dendritic cells to produce RANTES . Specific binding of CD40-transfected HEK 293 cells to Mtb HSP70 was demonstrated by surface plasmon resonance . Coimmunoprecipitation of Mtb HSP70 with CD40 indicates a physical association between these molecules . The results suggest that CD40 is critical in microbial HSP70 binding and stimulation of RANTES production.

J Agric Food Chem, 2002 Jan 2, 50(1), 136 - 9
Microbial response to bensulfuron-methyl treatment in soil; El-Ghamry AM et al.; A laboratory incubation study was conducted to evaluate the effect of bensulfuron-methyl treatment on soil microbial biomass and N-mineralization of a loamy sand soil . The herbicide was applied at 0 (control), 0.01 (field rate), 0.1, and 1.0 microg g(-1), and soil microbial biomass carbon (C(mb)), soil microbial biomass nitrogen (N(mb)), and N-mineralization rate (k) were measured at different times after herbicide treatment . Compared to the untreated soil, C(mb) and N(mb) decreased significantly (p < or = 0.05) within the first 7 days after herbicide treatment at 0.1 and 1.0 microg g(-1), and the impact was greater for N(mb) than for C(mb) . Nitrogen mineralization was significantly suppressed during the first 5 days of incubation when the soil was treated with bensulfuron-methyl at 0.1 and 1.0 microg g(-1) . The overall impact of bensulfuron-methyl to the soil microbial communities was closely related to the application rate in the range of 0.01-1.0 microg g(-1) . This effect, however, was found to be transitory, and significant impact occurred only at high application rates.

Nucleic Acids Res, 2002 Jan 1, 30(1), 66 - 8
Genome Information Broker (GIB): data retrieval and comparative analysis system for completed microbial genomes and more; Fumoto M et al.; Genome Information Broker (GIB) is a powerful tool for the study of comparative genomics . GIB allows users to retrieve and display partial and/or whole genome sequences together with the relevant biological annotation . GIB has accumulated all the completed microbial genome and has recently been expanded to include Arabidopsis thaliana genome data from DDBJ/EMBL/GenBank . In the near future, hundreds of genome sequences will be determined . In order to handle such huge data, we have enhanced the GIB architecture by using XML, CORBA and distributed RDBs . We introduce the new GIB here . GIB is freely accessible at http://gib.genes.nig.ac.jp/.

Nucleic Acids Res, 2002 Jan 1, 30(1), 27 - 30
DNA Data Bank of Japan (DDBJ) for genome scale research in life science; Tateno Y et al.; The DNA Data Bank of Japan (DDBJ, has made an effort to collect as much data as possible mainly from Japanese researchers . The increase rates of the data we collected, annotated and released to the public in the past year are 43% for the number of entries and 52% for the number of bases . The increase rates are accelerated even after the human genome was sequenced, because sequencing technology has been remarkably advanced and simplified, and research in life science has been shifted from the gene scale to the genome scale . In addition, we have developed the Genome Information Broker (GIB, that now includes more than 50 complete microbial genome and Arabidopsis genome data . We have also developed a database of the human genome, the Human Genomics Studio (HGS, HGS provides one with a set of sequences being as continuous as possible in any one of the 24 chromosomes . Both GIB and HGS have been updated incorporating newly available data and retrieval tools.

J Immunol, 2002 Jan 1, 168(1), 434 - 42
Bacterial lipopolysaccharide selectively up-regulates the function of the chemotactic peptide receptor formyl peptide receptor 2 in murine microglial cells; Cui YH et al.; Receptors for the bacterial chemotactic peptide fMLP are implicated in inflammation and host defense against microbial infection . We investigated the expression and function of fMLPR in microglial cells, which share characteristics of mononuclear phagocytes and play an important role in proinflammatory responses in the CNS . The expression of the genes encoding formyl peptide receptor (FPR)1 and FPR2, the high- and low-affinity fMLPR, was detected in a murine microglial cell line N9, but these cells did not respond to chemotactic agonists known for these receptors . N9 cells incubated with bacterial LPS increased the expression of fMLPR genes and developed a species of specific, but low-affinity, binding sites for fMLP, in association with marked calcium mobilization and chemotaxis responses to fMLP in a concentration range that typically activated the low-affinity receptor FPR2 . In addition, LPS-treated N9 cells were chemoattracted by two FPR2-specific agonists, the HIV-1 envelope-derived V3 peptide, and the 42 aa form of the amyloid beta peptide which is a pathogenic agent in Alzheimer's disease . Primary murine microglial cells also expressed FPR1 and FPR2 genes, but similar to N9 cells, exhibited FPR2-mediated activation only after LPS treatment . In contrast to its effect on the function of FPR2, LPS reduced N9 cell binding and biological responses to the chemokine stromal cell-derived factor-1alpha . Thus, LPS selectively modulates the function of chemoattractant receptors in microglia and may promote host response in inflammatory diseases in the CNS.

J Immunol, 2002 Jan 1, 168(1), 51 - 6
Immunostimulatory DNA sequences influence the course of adjuvant arthritis; Ronaghy A et al.; Bacterial DNA is enriched in unmethylated CpG motifs that have been shown to activate the innate immune system . These immunostimulatory DNA sequences (ISS) induce inflammation when injected directly into joints . However, the role of bacterial DNA in systemic arthritis is not known . The purpose of the present experiments was to determine whether ISS contributes to the development of adjuvant arthritis in Lewis rats after intradermal injection of heat-killed Mycobacterium tuberculosis (Mtb) . The results showed that Mtb DNA was necessary for maximal joint inflammation in adjuvant arthritis but could be replaced by synthetic ISS oligodeoxynucleotides . The arthritis-promoting effect of the Mtb DNA or of the ISS oligodeoxynucleotides correlated with an increased Th1 response to Mtb Ags, as measured by the production of IFN-gamma and increased production of the osteoclast differentiation factor, receptor activator of NF-kappaB ligand (RANKL) . The Mtb DNA did not enter the joints but dispersed to the bone marrow and spleen before the onset of systemic joint inflammation . Thus, adjuvant arthritis is a microbial DNA-dependent disease . In this model, we postulate that massive and prolonged activation of macrophages, dendritic cells, and osteoclast precursors in the bone marrow may prime the joints for the induction of inflammatory Th1 immune responses to Mtb Ags.

J Immunol, 2002 Jan 1, 168(1), 44 - 50
Generation of diversity in the innate immune system: macrophage heterogeneity arises from gene-autonomous transcriptional probability of individual inducible genes; Ravasi T et al.; Microbial products such as LPS stimulate macrophages to produce a wide diversity of inducible gene products needed for immediate host defense and priming of an appropriate acquired immune response . In this study, we have examined LPS-inducible gene expression in subclones of a mouse macrophage cell line, RAW264, using cDNA microarrays . Even archetypal target genes such as TNF-alpha were not induced in all subclones, and there was no absolute correlation between expression of pairs of genes . Nevertheless, the array analysis revealed clusters of genes that were more likely to be coexpressed . RAW264 cells stably transfected with luciferase reporter genes driven by LPS-responsive promoters revealed the same kind of clonal heterogeneity . The results indicate that each LPS-inducible gene has its own inherent probability of activation in response to LPS.

Clin Chem, 2002 Jan, 48(1), 121 - 30
Antigen microarrays for serodiagnosis of infectious diseases; Mezzasoma L et al.; BACKGROUND: Progress in robotic printing technology has allowed the development of high-density nucleic acid and protein arrays that have increased the throughput of a variety of assays . We generated protein microarrays by printing microbial antigens to simultaneously determine in human sera antibodies directed against Toxoplasma gondii, rubella virus, cytomegalovirus (CMV), and herpes simplex virus (HSV) types 1 and 2 (ToRCH antigens) . METHODS: The antigens were printed on activated glass slides with high-speed robotics . The slides were incubated first with serum samples and subsequently with fluorescently labeled secondary antibodies . Human IgG and IgM bound to the printed antigens were detected by confocal scanning microscopy and quantified with internal calibration curves . Both microarrays and commercial ELISAs were utilized to detect serum antibodies against the ToRCH antigens in a panel of characterized human sera . RESULTS: The detection limit (mean + 2 SD) of the microarray assay was 0.5 pg of IgG or IgM bound to the slides . Within-slide, between-slide, and between-batch precision profiles showed CVs of 1.7-18% for all antigens . Overall, >80% concordance was obtained between microarray assays and ELISAs in the classification of sera; for T . gondii, CMV, and HSV1, concordance exceeded 90% . CONCLUSIONS: The microarray is a suitable assay format for the serodiagnosis of infectious diseases and can be easily optimized for clinical use . The ToRCH assay performs equivalently to ELISA and may have potentially important advantages in throughput, convenience, and cost.

Biochim Biophys Acta, 2001 Nov 30, 1534(1), 34 - 44
Lipase-catalysed hydrolysis of short-chain substrates in solution and in emulsion: a kinetic study; Nini L et al.; We have studied the enzymatic hydrolysis of solutions and emulsions of vinyl propionate, vinyl butyrate and tripropionin by lipases of various origin and specificity . Kinetic studies of the hydrolysis of short-chain substrates by microbial triacylglycerol lipases from Rhizopus oryzae, Mucor miehei, Candida rugosa, Candida antarctica A and by (phospho)lipase from guinea-pig pancreas show that these lipolytic enzymes follow the Michaelis-Menten model . Surprisingly, the activity against solutions of tripropionin and vinyl esters ranges from 70% to 90% of that determined against emulsions . In contrast, a non-hyperbolic (sigmoidal) dependence of enzyme activity on ester concentration is found with human pancreatic lipase, triacylglycerol lipase from Humicola lanuginosa (Thermomyces lanuginosa) and partial acylglycerol lipase from Penicillium camembertii and the same substrates . In all cases, no abrupt jump in activity (interfacial activation) is observed at substrate concentration corresponding to the solubility limit of the esters . Maximal lipolytic activity is always obtained in the presence of emulsified ester . Despite progress in the understanding of structure-function of lipases, interpretation of the mode of action of lipases active against solutions of short-chain substrates remains difficult . Actually, it is not known whether these enzymes, which possess a lid structure, are in open or/and closed conformation in the bulk phase and whether the opening of the lid that gives access to the catalytic triad is triggered by interaction of the enzyme molecule with monomeric substrates or/and multimolecular aggregates (micelles) both present in the bulk phase . From the comparison of the behaviour of lipases used in this study which, in some cases, follow the Michaelis-Menten model and, in others, deviate from classical kinetics, it appears that the activity of classical lipases against soluble short-chain vinyl esters and tripropionin depends not only on specific interaction with single substrate molecules at the catalytic site of the enzyme but also on physico-chemical parameters related to the state of association of the substrate dispersed in the aqueous phase . It is assumed that the interaction of lipase with soluble multimolecular aggregates of tripropionin or short-chain vinyl esters or the formation of enzyme-substrate mixed micelles with ester bound to lipase, might represent a crucial step that triggers the structural transition to the open enzyme conformation by displacement of the lid.

FEMS Microbiol Lett, 2001 Dec 18, 205(2), 237 - 42
Comparative phylogeny of the ammonia monooxygenase subunit A and 16S rRNA genes of ammonia-oxidizing bacteria; Aakra A et al.; A fragment of the ammonia monooxygenase gene (amoA) from 31 strains of ammonia-oxidizing bacteria (AOB) was sequenced and analysed phylogenetically . The results were compared with the phylogeny of 16S rDNA from AOB . For most groups of AOB we found a high consistency between the phylogenetic trees based on the 16S rDNA and amoA sequences . Although it is not a phylogenetic marker, using the amoA as a probe when studying microbial diversity will probably reduce the amount of non-AOB detected, compared to using rDNA based probes . The data presented in this paper extend and improve the basis for application of amoA in studies of AOB in the environment.

Biomacromolecules, 2001 Spring, 2(1), 105 - 10
Effect of transglutaminase on reconstruction and physicochemical properties of collagen gel from shark type I collagen; Nomura Y et al.; The effects of microbial transglutaminase (MTGase) on type I collagen self-assembly and properties of reconstructed collagen fibrils from shark were investigated . Collagen self-assembly was accelerated with the addition of MTGase in dependence on that concentration . The relative amount of reconstructed collagen slightly decreased with MTGase . The diffusion coefficient of collagen gel was reduced by treatment with MTGase, and that suggested the reduction of mobility of the whole collagen network . At a high temperature, used to denature the collagen, MTGase-treated collagen gel remained as aggregates . By differential scanning calorimetry, the denaturation temperature of MTGase-treated collagen gel was about 2 degrees C higher than that of nontreated collagen gel . Treatment with MTGase yielded thermally stable cross-links in collagen molecules.

Chirality, 2002 Jan, 14(1), 51 - 8
Absolute configuration of 3-hydroxy acids formed by Stenotrophomonas maltophilia: application of multidimensional gas chromatography and circular dichroism spectroscopy; Weil K et al.; The soil bacterium Stenotrophomonas maltophilia was found to transform various long-chain fatty acids selectively into 3-hydroxy fatty acids of shorter chain length . Their chiral evaluation was performed by multidimensional gas chromatography (MDGC) on modified cyclodextrin phase comparing the enantiodistribution of 1,3-diol formed without loss of stereochemical information from a representative microbial product with those of synthetic (3RS)- and (3S)-1,3-diols . Enantiomeric excesses of 84-98% (R) were determined for the microbially produced 3-hydroxy acids . In addition, the CD exciton chirality method was applied to determine their absolute configuration . Derivatization with 9-anthryldiazomethane and 2-naphthoylimidazole led to the required bichromophoric structures . Their CD spectra displayed a positive first Cotton effect around 254 nm and a negative second Cotton effect around 237 nm, which confirmed the (R)-configuration of the bacterial products .

Circulation, 2001 Dec 18, 104(25), 3103 - 8
Toll-like receptor-4 is expressed by macrophages in murine and human lipid-rich atherosclerotic plaques and upregulated by oxidized LDL; Xu XH et al.; BACKGROUND: Inflammation is implicated in atherogenesis and plaque disruption . Toll-like receptor 2 (TLR-2) and TLR-4, a human homologue of drosophila Toll, play an important role in the innate and inflammatory signaling responses to microbial agents . To investigate a potential role of these receptors in atherosclerosis, we assessed the expression of TLR-2 and TLR-4 in murine and human atherosclerotic plaques . METHODS AND RESULTS: Aortic root lesions of high-fat diet-fed apoE-deficient mice (n=5) and human coronary atherosclerotic plaques (n=9) obtained at autopsy were examined for TLR-4 and TLR-2 expression by immunohistochemistry . Aortic atherosclerotic lesions in all apoE-deficient mice expressed TLR-4, whereas aortic tissue obtained from control C57BL/6J mice showed no TLR-4 expression . All 5 lipid-rich human plaques expressed TRL-4, whereas the 4 fibrous plaques and 4 normal human arteries showed no or minimal expression . Serial sections and double immunostaining showed TLR-4 colocalizing with macrophages both in murine atherosclerotic lesions and at the shoulder region of human coronary artery plaques . In contrast to TLR-4, none of the plaques expressed TLR-2 . Furthermore, basal TLR-4 mRNA expression by human monocyte-derived macrophages was upregulated by ox-LDL in vitro . CONCLUSIONS: Our study demonstrates that TLR-4 is preferentially expressed by macrophages in murine and human lipid-rich atherosclerotic lesions, where it may play a role to enhance and sustain the innate immune and inflammatory responses . Moreover, upregulation of TLR-4 in macrophages by oxidized LDL suggests that TLR-4 may provide a potential pathophysiological link between lipids and infection/inflammation and atherosclerosis.

Emerg Infect Dis, 2001 Sep-Oct, 7(5), 820 - 7
Clinical consequences and cost of limiting use of vancomycin for perioperative prophylaxis: example of coronary artery bypass surgery; Zanetti G et al.; Routine us of vancomycin for perioperative prophylaxis is discouraged, principally to minimize microbial resistance to it . However, outcomes and costs of this recommendation have not been assessed . We used decision-analytic models to compare clinical results and cost-effectiveness of no prophylaxis, cefazolin, and vancomycin, in coronary artery bypass graft surgery . In the base case, vancomycin resulted in 7% fewer surgical site infections and 1% lower all-cause mortality and saved $117 per procedure, compared with cefazolin . Cefazolin, in turn, resulted in substantially fewer infections and deaths and lower costs than no prophylaxis . We conclude that perioperative antibiotic prophylaxis with vancomycin is usually more effective and less expensive than cefazolin . Data on vancomycin's impact on resistance are needed to quantify the trade-off between individual patients' improved clinical outcomes and lower costs and the future long-term consequences to society.

J Pharm Sci, 2001 Nov, 90(11), 1787 - 94
Synthesis and evaluation of N-acyl-2-(5-fluorouracil-1-yl)-D,L-glycine as a colon-specific prodrug of 5-fluorouracil; Lee JS et al.; N-nicotinyl-2-(5-fluorouracil-1-yl)-D,L-glycine (NFG) and N-isonicotinyl-2-(5-fluorouracil-1-yl)-D,L-glycine (INFG) were synthesized as colon-specific prodrugs of 5-fluorouracil (5-FU) . As N-aromatic acyl amides of glycine, they are expected to be stable in the upper intestine and delivered to the colon as an intact form if they are nonabsorbable . Microbial hydrolysis of amide bond in the colon will give 2-(5-fluorouracil)-D,L-glycine, which releases 5-FU by spontaneous decomposition . NFG and INFG were soluble in water and stable in pH 1.2 and 7.4 buffer solutions . The apparent partition coefficient of NFG or INFG in 1-octanol/pH 7.4 phosphate buffer solution at 37 degrees was 0.025 or 0.024, respectively . On incubation with cecal contents of rats, conversion of NFG or INFG proceeded only 9 or 5% in 8 h, respectively, producing 5-FU and a metabolite . The metabolite formation was inhibited in the presence of diazouracil, a dihydrouracil dehydrogenase inhibitor . The HPLC retention time of the metabolite from the incubation of 5-FU, NFG, or INFG with cecal contents was identical to dihydro-5-fluorouracil (dihydro-5FU) . When N-nicotinyl-2-hydroxy-D,L-glycine methyl ester (NHGM) was incubated with the cecal contents, the extent of amide bond hydrolysis was 85% in 24 h . The result suggested that steric hindrance imposed by 5-FU at 2-position of glycine retarded the hydrolysis of the amide bond in NFG or INFG and suppressed the prodrug conversion .

J Biomed Mater Res, 2002 Jan, 59(1), 176 - 83
Mutagenicity evaluation of forty-one metal salts by the umu test; Yamamoto A et al.; Metallic biomaterials implanted in a human body may corrode and wear, releasing metal ions and debris which may induce adverse reactions such as inflammation, allergy, neoplastic formation, developmental malformation, etc . Mutagenicity is a very fundamental and important toxicity related to carcinogenicity and reproductive/developmental toxicity because the damages to genes or DNA can be a cause of carcinogenesis and developmental abnormalities . However, available mutagenic data on metallic ions and compounds are restricted to the number of elements . Therefore, to obtain the systematic data necessary for metal ion mutagenicity, 41 metal salts encompassing 36 metals and 5 metallic elements tested with different valences, were evaluated on their mutagenicity by a microbial test, the umu test . As a result, K(2)Cr(2)O(7), RhCl(3), IrCl(4), and MgCl(2) are positive without metabolic activation . Concentrations having the maximum mutagenic effect (C(max)) are 9.65 x 10(-5), 1.00 x 10(-4), 3.11 x 10(-3), 4.12 x 10(-3) mol . L(-1), respectively . CuCl(2), VCl(3), CuCl, RhCl(3), K(2)Cr(2)O(7), and IrCl(4) are positive with metabolic activation by S-9 mix with C(max) of 1.60 x 10(-5), 3.91 x 10(-5), 1.57 x 10(-4), 2.00 x 10(-4), 3.86 x 10(-4), 1.56 x 10(-2) mol . L(-1), respectively . Thirty-five metal salts were negative for tests performed both with and without metabolic activation, whereas it was impossible to evaluate the mutagenicity of MoCl(5) and ZrCl(4) by the umu test because of their colorimetric reaction to testing reagents .

Eur J Immunol, 2001 Dec, 31(12), 3525 - 34
Distinct CpG oligonucleotide sequences activate human gamma delta T cells via interferon-alpha/-beta; Rothenfusser S et al.; Oligodeoxynucleotides with CpG motifs (CpG ODN) mimic microbial DNA and activate effectors of innate immunity including NK cells . Human gamma delta T cells (Vgamma9/Vdelta2) are antigen specific "natural memory" T cells in a preactivated stage, which respond to common non-protein phosphoantigens . Among several CpG ODN tested, distinct CpG ODN sequences characterized by inducing high amounts of IFN-alpha/-beta in PBMC elicited strong gamma delta T cell and NK cell responses, as determined by CD69 expression, IFN-gamma production, perforin content and lytic activity . These CpG ODN activated gamma delta T cells and NK cells in the absence of an additional stimulus and synergistically increased responsiveness to cell-type-specific antigens like isopentenylpyrophosphate for gamma delta T cells and NK-sensitive tumor cells for NK cells . NK cells and gamma delta T cells were activated via IFN-alpha/-beta released by CpG ODN-stimulated PBMC . Purified gamma delta T cells and NK cells did not respond to CpG ODN but to recombinant IFN-alpha/-beta . In conclusion, CpG ODN sequences were identified which, based on their ability to induce high amounts of IFN-alpha/-beta, represent strong adjuvants for "natural memory" cells including responses of gamma delta T cells to non-protein antigens . Early IFN-alpha/-beta dependent stimulation of IFN-gamma synthesis in NK cells and gamma delta T cells may contribute to the CpG ODN-induced Th1 bias of an evolving immune response.

Eur J Immunol, 2001 Nov, 31(11), 3388 - 93
Specialization and complementarity in microbial molecule recognition by human myeloid and plasmacytoid dendritic cells; Jarrossay D et al.; Following encounter with pathogens, dendritic cells (DC) mature and migrate from peripheral tissues to the T cell areas of secondary lymphoid organs, where they produce regulatory cytokines and prime naive T lymphocytes . We investigated in two subsets of human peripheral blood DC the expression of Toll-like receptors (TLR1 through TLR9) and the regulation of chemokine receptors and cytokine production in response to different maturation stimuli . Myeloid DC express all TLR except TLR7 and TLR9, which are selectively expressed by plasmacytoid DC . Myeloid and plasmacytoid DC respond to pathogen-associated molecular patterns according to their TLR expression . In response to the appropriate stimuli both DC types up-regulate CCR7, a receptor that drives DC migration to the T cell areas . Type I IFN was produced only by plasmacytoid DC and at early time points after stimulation . Furthermore, its production was elicited by some of the maturation stimuli tested . These results reveal a remarkable specialization and complementarity in microbial molecule recognition as well as a flexibility in effector function among myeloid and plasmacytoid DC.

Biotechnol Bioeng, 2001 Dec 5, 75(5), 607 - 14
Significance of location of enzymes on their release during microbial cell disruption; Balasundaram B et al.; The release kinetics of the enzyme invertase and alcohol dehydrogenase from yeast and penicillin acylase from E . coli during disruption using various techniques has been investigated . The disruption techniques used were sonication, high-pressure homogenization, and hydrodynamic cavitation . The first-order-release kinetics was applied for the determination of release rate of these enzymes and total soluble proteins . Location factor (LF) values were calculated using these release rates . The location of the enzymes as given by the values of location factor coincided well with those reported in the literature . Varying values of location factor for the same enzyme by different disruption techniques gave some indications about the selectivity of release of a target enzyme by different disruption techniques . Varying values of location factor for the same enzyme with the use of a particular equipment or disruption technique at different conditions reveals the degree to which the cell is disrupted . Few plausible applications of this location factor concept have been predicted and these speculations have been examined . This location factor concept has been used for monitoring the heat-induced translocation of ADH and location of penicillin acylase during the growth period of E . coli cells .

J Nippon Med Sch, 2001 Dec, 68(6), 466 - 71
{CD1: A new paradigm for antigen presentation}; Sugita M; Molecules of the major histocompatibility complex (MHC) bind protein-derived peptide antigens and present them to T cells . This has been a central dogma in modern immunology, and our appreciation of a variety of cell-mediated immune responses has been based only on this paradigm . However, we now know that T cell recognition also involves non-peptide antigens . Studies over the past several years have established a new paradigm that non-MHC-encoded molecules of the CD1 family mediate presentation of lipid antigens to T cells, and unraveled their significant role in microbial immunity, tumor immunology, and autoimmunity . Identification of a novel pathway for T cell activation mediated by CD1 molecules opens a possibility for new therapeutic strategies, including development of lipid-based vaccines.

Environ Health Perspect, 2001 Dec, 109 Suppl 6, 877 - 84
Approaches to detecting immunotoxic effects of environmental contaminants in humans; Tryphonas H; Experimental animal studies indicate that environmental contaminants can have adverse effects on several organs and tissues of the immune system . Such effects are known to lead to increased host susceptibility to microbial infections and to compromised immunosurveillance mechanisms normally instrumental in the elimination of neoplastic cells and the prevention of autoimmune diseases . Evaluation of the potential risk environmental contaminants pose to the human immune system is currently accomplished via extrapolation of experimentally derived animal data to humans . Presently, this process requires that uncertainty factors such as interspecies differences and genetic variability be considered . Naturally, the process of risk assessment would be greatly facilitated if it were based on clinically relevant data derived from studying humans known to be exposed to environmental contaminants . However, the existing human data are scarce and often described as very limited in scope . To generate the much-needed human data we need to identify a set of clinically relevant immunologic end points that, when adequately standardized, can be incorporated easily into the design of prospective epidemiologic studies.

J Agric Food Chem, 2001 Dec, 49(12), 5778 - 84
Plant factors influencing enzyme retting of fiber and seed flax; Akin DE et al.; Retting, which is the microbial activity through which bast fibers are released from nonfiber tissues, is the limiting factor in flax processing . The objective of this work is to identify chemical and structural characteristics in a variety of fiber and seed flax types that influence enzyme retting in a recently developed method . Analyses of flax retted in a series of tests, including two enzyme rettings in some cases, indicated that lignin did not limit the separation of fibers from shive and showed that pectinases in enzyme-retting mixtures could ret fiber and seed flax . However, mature stems, such as that in flax produced for seed, had greater amounts of cutin and wax in the cleaned fiber product, suggesting that the cuticle could be a greater antiquality factor in seed versus fiber flax . With seed flax, the fraction of finer fibers produced during retting was significantly lower than with fiber flax . Results indicated that enzyme retting could be used to obtain flax fibers from seed flax stem residues and add value to this agricultural material.

Nat Immunol, 2002 Jan, 3(1), 76 - 82 Epub 2001 Dec 17.
Lipoxin-mediated inhibition of IL-12 production by DCs: a mechanism for regulation of microbial immunity; Aliberti J et al.; Lipoxins are eicosanoid mediators that show potent inhibitory effects on the acute inflammatory process . We show here that the induction of lipoxin A(4) (LXA(4)) accompanied the in vivo suppression of interleukin 12 (IL-12) responsiveness of murine splenic dendritic cells (DCs) after microbial stimulation with an extract of Toxoplasma gondii . This paralysis of DC function could not be triggered in mice that were deficient in a key lipoxygenase involved in LXA(4) biosynthesis . In addition, DCs pre-treated with LXA(4) became refractory to microbial stimulation for IL-12 production in vitro and mice injected with a stable LXA(4) analog showed reduced splenic DC mobilization and IL-12 responses in vivo . Together, these findings indicate that the induction of lipoxins in response to microbial stimulation can provide a potent mechanism for regulating DC function during the innate response to pathogens.

Mol Ecol, 2001 Oct, 10(10), 2409 - 22
Multilocus nested haplotype networks extended with DNA fingerprints show common origin and fine-scale, ongoing genetic divergence in a wild microbial metapopulation; Carbone I et al.; Nested haplotype networks for three loci in a haploid, fungal plant pathogen, Sclerotinia sclerotiorum, in two natural, Norwegian populations of the woodland buttercup, Ranunculus ficaria, were extended with DNA fingerprints to determine fine-scale population divergence . To preserve the cladistic structure in the network for both nonrecombinant and postrecombinant haplotypes in highly recombinant clades, recombinant events were not removed ('peeled off'), but instead were examined in alternative (marginal) networks . Fungi from both sampling locations share a common origin with subsequent genetic divergence, consistent with expectations for metapopulation structure . Evidence for divergence includes (i) lack of shared fingerprints between the two locations, (ii) evolution of new fingerprints, via transposition and recombination, within 2 years on a fine spatial scale within one sampling location, and (iii) increase in the size of the intergenic spacer (IGS) in both sampling locations . Sites of microsatellite repeat expansion and of an insertion were consistent with the boundaries of two recombination blocks in the IGS . Both alternative networks based on the recombination blocks were essential to finding all associations of DNA fingerprints with IGS size, sampling site, sampling year and mycelial compatibility group . Variation in the elongation factor 1alpha and calmodulin loci supported the topologies and the recurrent, ongoing polarity of change in fingerprints and IGS size inferred from the IGS.

J Bacteriol, 2002 Jan, 184(1), 91 - 103
ATP-binding cassette transport system involved in regulation of morphological differentiation in response to glucose in Streptomyces griseus; Seo JW et al.; Streptomyces griseus NP4, which was derived by UV mutagenesis from strain IFO13350, showed a bald and wrinkled colony morphology in response to glucose . Mutant NP4 formed ectopic septa at intervals along substrate hyphae, and each of the compartments developed into a spore which was indistinguishable from an aerial spore in size, shape, and thickness of the spore wall and in susceptibility to lysozyme and heat . The ectopic spores of NP4 formed in liquid medium differed from "submerged spores" in lysozyme sensitivity . Shotgun cloning experiments with a library of the chromosomal DNA of the parental strain and mutant NP4 as the host gave rise to DNA fragments giving two different phenotypes; one complementing the bald phenotype of the host, and the other causing much severe wrinkled morphology in the host . Subcloning identified a gene (dasR) encoding a transcriptional repressor belonging to the GntR family that was responsible for the reversal of the bald phenotype and a gene (dasA) encoding a lipoprotein probably serving as a substrate-binding protein in an ATP-binding cassette (ABC) transport system that was responsible for the severe wrinkled morphology . These genes were adjacent but divergently encoded . Two genes, named dasB and dasC, encoding a membrane-spanning protein were present downstream of dasA, which suggested that dasRABC comprises a gene cluster for an ABC transporter, probably for sugar import . dasR was transcribed actively during vegetative growth, and dasA was transcribed just after commencement of aerial hypha formation and during sporulation, indicating that both were developmentally regulated . Transcriptional analysis and direct sequencing of dasRA in mutant NP4 suggested a defect of this mutant in the regulatory system to control the expression of these genes . Introduction of multicopies of dasA into the wild-type strain caused ectopic septation in very young substrate hyphae after only 1 day of growth and subsequent sporulation in response to glucose . The ectopic spores of the wild type had a thinner wall than those of mutant NP4, in agreement with the observation that the former was sensitive to lysozyme and heat . Disruption of the chromosomal dasA or dasR in the wild-type strain resulted in growth as substrate mycelium, suggesting an additional role of these genes in aerial mycelium formation . The ectopic septation and sporulation in mutant NP4 and the wild-type strain carrying multicopies of dasA were independent of a microbial hormone, A-factor (2-isocapryloyl-3R-hydroxymethyl-gamma-butyrolactone), that acts as a master switch of aerial mycelium formation and secondary metabolism.

J Hosp Infect, 2001 Dec, 49(4), 268 - 73
Bacterial contamination of blenderized whole food and commercial enteral tube feedings in the Philippines; Sullivan MM et al.; Hospital-prepared tube feedings from four Philippine acute-care hospitals were analysed for microbial contamination . Two feedings were prepared on three separate days at each hospital . The tube feedings were either blended natural whole foods or a reconstituted commercial powdered nutritional product . Samples of each feeding were collected for coliform count and standard plate count at the time of preparation and at 1, 2 and 4 h after preparation after maintenance at room temperature (26-31 degrees C).At the time of preparation, mean coliform and standard plate counts for all samples were 10.3 most probable number per gram (MPN/g) and 7.4x10(4)colony-forming units per gram (cfu/g), respectively . Nine of 24 samples (38%) had coliform counts greater than 10 MPN/g, and 22/24 (92%) samples had standard plate counts greater than 10(3) cfu/g . There were significant increases in mean coliform and standard plate counts over 4 h (P=0.0005 and P=0.008, respectively) . At 4 h after preparation, the mean coliform and standard plate counts were 18.2 MPN/g and 2.1x10(5) cfu/g, respectively . At this time, 18/24 (75%) samples had coliform counts greater than 10 MPN/g and 20/24 (83%) samples had standard plate counts greater than 10(5) cfu/g . The results of this study show that the microbial quality of the majority of the hospital-prepared enteral tube feedings analysed were not within published guidelines for safety .

J Immunol, 2001 Dec 15, 167(12), 6773 - 9
Essential contribution of germline-encoded lysine residues in Jgamma1.2 segment to the recognition of nonpeptide antigens by human gammadelta T cells; Miyagawa F et al.; Human gammadelta T cells display unique repertoires of Ag specificities largely imposed by selective usages of distinct Vgamma and Vdelta genes . Among them, Vgamma2/Vdelta2(+) T cells predominate in the circulation of healthy adults and respond to various microbial small molecular mass nonpeptide Ags . The present results indicate that the primary Vgamma2/Vdelta2(+) T cells stimulated with the distinct groups of nonpeptide Ags, including monoethyl pyrophosphate, isobutyl amine, and aminobisphosphonate, invariably exhibit Jgamma1.2 in the Vgamma2(+) TCR-gamma chains . Gene transfer studies revealed that most of the randomly cloned Vgamma2/Jgamma1.2(+) TCR-gamma genes bearing diverse Vgamma/Jgamma junctional sequences could confer the responsiveness to all these nonpeptide Ags, while none of the Vgamma2/Jgamma1.1(+) or Vgamma2/Jgamma1.3(+) TCR-gamma genes could do so . Furthermore, mutation of the lysine residues encoded by the Jgamma1.2 gene, which are unique in human Jgamma1.2 and absent in other human or mouse Jgamma segments, completely abrogated the responsiveness to all the nonpeptide Ags without affecting the response to anti-CD3 mAb . These results strongly suggested that the positively charged lysine residues in the TCR-gamma chain CDR3 region encoded by the germline Jgamma1.2 gene play a key role in the recognition of diverse small molecular mass nonpeptide Ags.

BMC Biotechnol . 2001;1(1):10 . Epub 2001 Nov 20.
Long term adaptation of a microbial population to a permanent metabolic constraint: overcoming thymineless death by experimental evolution of Escherichia coli; de Crecy-Lagard VA et al.; BACKGROUND: To maintain populations of microbial cells under controlled conditions of growth and environment for an indefinite duration is a prerequisite for experimentally evolving natural isolates of wild-type species or recombinant strains . This goal is beyond the scope of current continuous culture apparatus because these devices positively select mutants that evade dilution, primarily through attachment to vessel surfaces, resulting in persistent sub-populations of uncontrollable size and growth rate . RESULTS: To overcome this drawback, a device with two growth chambers periodically undergoing transient phases of sterilization was designed . The robustness of this device was assessed by propagating an E . coli strain under permanent thymine starvation for over 880 days, i.e . metabolic conditions notoriously known to lead to cell death and clogging of cultivation vessels . Ten thousand generations were required to obtain a descendant lineage that could resist thymine starvation and had recovered wild-type growth rate . CONCLUSIONS: This approach provides a technological framework for the diversification and improvement of microbial strains by long-term adaptation to inescapable metabolic constraints . An E . coli strain that is totally resistant to thymineless death was selected.

Clin Exp Allergy, 2001 Dec, 31(12), 1839 - 45
Pets and vermin are associated with high endotoxin levels in house dust; Heinrich J et al.; BACKGROUND: Previous studies have shown that the risk for allergic sensitization is lower in children who grew up on farms and in young adults who were exposed to dogs in early childhood . A higher microbial exposure in general and in particular to endotoxin in early childhood might contribute to this lower risk of atopy . OBJECTIVE: We examined whether the presence of pets or vermin in the home is associated with higher endotoxin concentrations in settled house dust . METHODS: House dust was sampled in a standardized manner on the living room floors of 454 homes of German children aged 5-10 years (participation rate 61%) . Endotoxin was assessed with a quantitative kinetic chromogenic Limulus Amebocyte Lysate (LAL) method . Associations between endotoxin levels, pets and vermin are presented as ratios of the crude and confounder adjusted geometric means (means ratios) in the category of study vs . a reference category using multiple linear regression models . RESULTS: Endotoxin concentrations in living room floor dust sampled in homes without pets and vermin were lower (1246 ng per square meter, 1519 ng endotoxin/g dust, n = 157) than those sampled in homes with pets or vermin (2267 ng per square meter, 2200 ng endotoxin/g dust, n = 296) . After adjustment for city of residence, season of dust sampling, age of the building and story of the dwelling, means ratios for endotoxin expressed per gram of dust were statistically significantly increased for dog (1.64, 95% CI 1.09-2.46), for cat (1.50, 95% CI 1.03-2.18) and for cockroach (3.01, 95% CI 1.37-6.60), whereas no major statistically significant associations were found for other pets, ants and mice . CONCLUSION: Keeping a dog or a cat in the home is consistent with higher exposure to endotoxin and might therefore contribute to the lower risk of atopy in later life.

Biotechnol Prog, 2001 Nov-Dec, 17(6), 1061 - 4
Rice straw as a support for immobilization of microbial lipase; de Castro HF et al.; Candida rugosa lipase was covalently immobilized on rice straw activated with glutaraldehyde using poly(ethylene glycol) (PEG) as the stabilizing agent . The effects of PEG molecular weight and enzyme loading were studied according to a full 2(2) factorial design . Higher immobilization yields (>70%) were attained when the lipase loading was 95 units/mg of dry support, independent of PEG molecular weight . All derivatives showed high hydrolytic and synthetic activities . This work provides preliminary results on the use of agricultural residues as a support matrix for immobilizing lipase and on the application of the resulting derivatives to butyl butyrate synthesis as a study model.

Eur J Biochem, 2001 Dec, 268(23), 6190 - 6
Arginine 121 is a crucial residue for the specific cytotoxic activity of the ribotoxin alpha-sarcin; Masip M et al.; Alpha-sarcin, a cyclizing ribonuclease secreted by the mould Aspergillus giganteus, is one of the best characterized members of a family of fungal ribotoxins . This protein induces apoptosis in tumour cells due to its highly specific activity on ribosomes . Fungal ribotoxins display a three-dimensional protein fold similar to those of a larger group of microbial noncytotoxic RNases, represented by RNases T1 and U2 . This similarity involves the three catalytic residues and also the Arg121 residue, whose counterpart in RNase T1, Arg77, is located in the vicinity of the substrate phosphate moiety although its potential functional role is not known . In this work, Arg121 of alpha-sarcin has been replaced by Gln or Lys . These two mutations do not modify the conformation of the protein but abolish the ribosome-inactivating activity of alpha-sarcin . In addition, the loss of the positive charge at that position produces dramatic changes on the interaction of alpha-sarcin with phospholipid membranes . It is concluded that Arg121 is a crucial residue for the characteristic cytotoxicity of alpha-sarcin and presumably of the other fungal ribotoxins.

Biochim Biophys Acta, 2001 Nov 29, 1537(3), 204 - 10
Risk of iron overload is decreased in beating heart coronary artery surgery compared to conventional bypass; Mumby S et al.; Conventional cardiopulmonary bypass surgery (CCPB) increases the iron loading of plasma transferrin often to a state of plasma iron overload, with the presence of low molecular mass iron . Such iron is a potential risk factor for oxidative stress and microbial virulence . Here we assess 'off-pump' coronary artery surgery on the beating heart for changes in plasma iron chemistry . Seventeen patients undergoing cardiac surgery using the 'Octopus' myocardial wall stabilisation device were monitored at five time points for changes in plasma iron chemistry . This group was further divided into those (n=9) who had one- or two- (n=8) vessel grafts, and compared with eight patients undergoing conventional coronary artery surgery . Patients undergoing beating heart surgery had significantly lower levels of total plasma non-haem iron, and a decreased percentage saturation of their transferrin at all time points compared to conventional bypass patients . Plasma iron overload occurred in only one patient undergoing CCPB . Beating heart surgery appears to decrease red blood cell haemolysis, and tissue damage during the operative procedures and thereby significantly decreases the risk of plasma iron overload associated with conventional bypass.

FEMS Microbiol Lett, 2001 Nov 13, 204(2), 335 - 40
Arsenite oxidation and arsenate respiration by a new Thermus isolate; Gihring TM et al.; A new microbial strain was isolated from an arsenic-rich terrestrial geothermal environment . The isolate, designated HR13, was identified as a Thermus species based on 16S rDNA phylogenetic relationships and close sequence similarity within the Thermus genus . Under aerobic conditions, Thermus HR13 was capable of rapidly oxidizing inorganic As(III) to As(V) . As(III) was oxidized at a rate approximately 100-fold greater than abiotic rates . Metabolic energy was not gained from the oxidation reaction . In the absence of oxygen, Thermus HR13 grew by As(V) respiration coupled with lactate oxidation . The ability to oxidize and reduce arsenic has not been previously described within the Thermus genus.

J Investig Med, 2001 Nov, 49(6), 514 - 21
Calcitonin gene-related peptide expression in sepsis: postulation of microbial infection-specific response elements within the calcitonin I gene promoter; Domenech VS et al.; BACKGROUND: Recently, we reported an unexpected ubiquitous expression of calcitonin (CT)-mRNA in a hamster peritonitis model of sepsis . Using this animal model,we undertook a study to further investigate the pattern of expression of the calcitonin I (CALC-I) gene and CT gene-related peptide (CGRP)-mRNA in sepsis . METHODS: Live Escherichia coli impregnated in agar pellets were implanted in the peritoneal cavities of hamsters . Twelve hours after sepsis induction, the septic and healthy control animals were sacrificed and tissues and peritoneal macrophages were collected . CGRP-mRNA content was evaluated by reverse transcription polymerase chain reaction (RT-PCR), quantitated by the Taq-Man technique, and compared with the mRNA expression of CT, tumor necrosis factor alpha (TNF-alpha), and interleukin-6 (IL-6) . The 5' untranslated regions of the mRNA and potential alternative splicing sites were identified by 5' rapid amplification of cDNA ends . RESULTS: We found a tissue-wide, ubiquitous and uniform expression of CGRP-mRNA in all septic tissues examined . CGRP-mRNA was detectable by RT-PCR in various extraneuronal and extrathyroidal septic tissues, but not in healthy control tissues . As found for CT-mRNA in our earlier studies, CGRP-mRNA seemed to be more specifically up-regulated as compared with other classical cytokines (ie, II-6 and TNF-alpha) . Importantly, the 5' untranslated sequence in control and septic thyroid was similar to the sequence obtained from septic spleen . CONCLUSIONS: We postulate the presence of microbial infection-specific response elements in the CALC-I gene promotor, which, upon a specific stimulus, override the tissue-selective expression pattern . This new form of endocrine plasticity may be of importance in the response to systemic inflammation.

Arch Immunol Ther Exp (Warsz), 1999, 47(1), 61 - 5
Effect of granulocyte colony stimulating factor treatment on ex vivo cytokine production by blood cells of patients after chemotherapy or radiotherapy; Kaminska T et al.; We explored ex vivo alterations in the cytokine release of stimulated blood cells taken from 8 patients with hematological malignancies who, after chemotherapy or radiotherapy developed leukopenia, and were treated for 3-7 days subcutaneously with granulocyte colony stimulating factor (G-CSF), daily, dose of 5 microg/kg of body weight . Blood was also taken from 8 healthy controls not treated with G-CSF and from patients before and 24 h after last dose of G-CSF and ex vivo treated with interferon (IFN) inducers: Newcastle disease virus (NDV), phytohemagglutinin (PHA), concanavalin A (Con A) and with tumor necrosis factor (TNF) inducer--lipopolysaccharide (LPS) . Blood cells of patients before G-CSF treatment exhibited ex vivo a low ability to produce IFN-gamma in comparison to controls . After G-CSF therapy a significant increase in IFN-alpha production ability was detected . We conclude that G-CSF treatment for 3-7 days does not only increase the number of white blood cells (WBC) and neutrophilic granulocytes but also modify the host response of patients with hematological malignancies to microbial infections.

Schweiz Monatsschr Zahnmed, 2001, 111(10), 1160 - 4
{Quality of water of dental units . A survey in the region of Berne, Switzerland}; Tonetti-Eberle B et al.; The aim of this study was to assess the microbial contamination of the water from 175 dental units, which had not been in use for at least 12 hours, and to determine the usefulness of rinsing the water lines for 3 minutes . Only 10% of all units fulfilled all criteria for drinking water . Frequently the total bacterial counts markedly exceeded 100 CFU/ml . In all cases the rinsing decreased bacterial loads significantly . Bacterial contamination of water lines increased with the time the dental units remained unused . It is recommended to rinse dental unit water lines daily for at least 3 minutes before working on patients.

Int Arch Allergy Immunol, 2001 Oct, 126(2), 102 - 10
How to deal with polarized Th2 cells: exploring the Achilles' heel; Smits HH et al.; The central effector cells in the pathogenesis of atopic allergic diseases are type 2 T helper (Th2) cells, which display an aberrant cytokine profile dominated by type 2 cytokines . Initial reports from mouse studies indicated that established and committed Th2 cells are stable and unsusceptible to modulation . However, there is a growing awareness that in humans, established effector Th2 cells are more flexible and can be reverted to predominant Th1 phenotypes . In fact, the Th1-driving cytokine interleukin (IL)-12 is the crucial factor in this respect . IL-12 is mainly produced by dendritic cells (DC), which can be primed for high or low IL-12 production, depending on inflammatory and/or microbial signals they encounter during their residence in the peripheral tissues . Accordingly, both the regulation of and the priming for IL-12 production in DC form ideal targets for therapeutic intervention . The development of new therapies for atopic allergy now focuses on local IL-12-promoting substances to target both the development of new Th2 cells and the persistent population of established allergen-specific Th2 cells .

Cell Mol Biol (Noisy-le-grand), 2001 Jul, 47(5), 865 - 83
Temperature effects on cell-functioning--a critical role for vicinal water; Drost-Hansen W; Interfacially modified water ('vicinal water') appears to extend over distances of several molecular diameters from an adjacent interface and possesses highly unusual thermal properties . Specifically, the vicinal water undergoes relatively abrupt changes over narrow temperature intervals centered around 15 degrees, 30 degrees, 45 degrees and 60 degrees C and these changes most likely reflect higher order (structural) phase transitions . Vicinal water occurs at all interfaces (independent of specific surface characteristics) and will therefore also be present in all biological cells . As a result, the effects of vicinal water ramify through all of cell biology and this explains many unexpected thermal anomalies seen in the temperature responses of a great variety of living systems . Thus, the concept of the thermal anomalies may explain such observations as abrupt changes in enzyme functioning; anomalous temperature effects in membrane processes; unusual cell volume responses to temperature; dramatic changes in erythrocyte sedimentation rates; anomalies in chromosome aberration rates; pasteurization temperature; upper and lower thermal limits for microbial growth; multiple growth optima and minima and body temperatures of mammals and birds.

Int J Med Microbiol, 2001 Nov, 291(5), 323 - 9
Dendritic cells as vectors for vaccination against infectious diseases; Moll H et al.; Antigen presentation by dendritic cells (DCs) is critical for the induction of a specific immune response . The immunotherapeutic potential of antigen-pulsed DCs for the treatment of cancer has been confirmed in a number of experimental tumor models and in several preclinical trials . Recent advances in our understanding of the interaction of microbial pathogens with DCs have provided the basis to explore DCs as vaccine carriers for the induction of protective immune responses to infections . Support for this strategy comes from animal studies demonstrating that DCs, after ex vivo loading with microbial antigens, confer protection against microbial challenges in vivo . This may have important implications for the development of novel strategies for prophylactic or therapeutic immunizations against various microbial pathogens.

Expert Opin Biol Ther, 2001 Jan, 1(1), 67 - 77
Heat shock proteins: novel therapeutic tools for HIV-infection?
Brenner BG, Wainberg Z.
Heat shock proteins (Hsps), cyclophilins (Cyps) and FK binding proteins (FKBPs) form a family of intracellular chaperone molecules that facilitate protein folding and assembly . These stress proteins are selectively expressed in cells in response to a range of stimuli, including heat, lymphokine and microbial/viral infections . This review discusses the role of stress proteins in the HIV-1 viral life cycle, with regard to the development of specific Hsp-based therapeutic strategies against HIV-1 infection . Cumulative findings are cited implicating CypA, Hsp27, Hsp70 and FKBPs in host cell and viral activation, viral entry, assembly or formation of infectious virions . Biological response modifiers that show specific high-affinity interactions with Cyp, FKBPs and Hsps, including cyclosporins, FK-506 and cyclopentenone prostaglandins respectively, may block HIV-1 replication and infection, providing novel HIV-1 therapeutic strategies . Moreover, Hsp binding to viral complexes can enhance antiviral immunity, including natural killer (NK), antibody-dependent (ADCC), gamma delta T-cell and cytotoxic T-lymphocyte (CTL) activities against HIV-1 infected cells . The ability of Hsps to interact with HIV-1 viral proteins, combined with their inherent adjuvant and immunogenic properties indicates that Hsps may also serve as vehicles for antigen delivery and the design of AIDS vaccines.

Expert Opin Biol Ther, 2001 Jul, 1(4), 641 - 53
Fighting infection using immunomodulatory agents; Masihi KN; The last decade has seen the emergence of immunomodulators as promising therapeutic agents in infectious diseases . A diverse array of recombinant, synthetic and natural immunomodulatory preparations for prophylaxis and treatment of various infections are available today . Some of these substances, such as granulocyte colony-stimulating factor (G-CSF), interferons, imiquimod and bacterial-derived preparations are already licensed for use in patients . Others including IL-12, various chemokines, synthetic cytosine phosphate-guanosine (CpG) oligodeoxynucleotides and glucans are being investigated extensively in clinical and preclinical studies . Immunomodulatory regimens offer an attractive approach as an adjunct modality for control of microbial diseases in the era of antibiotic resistance . Practical application of the advances in molecular biology, bioinformatics, genomic mining and high-throughput peptide synthesis should foster future discovery and development of novel immunomodulators contingent upon scientific evidence rather than dictates of discursive empiricism.

ESA Bull, 1999 Mar, 97(5), 56 - 60
Water recovery in space; Tamponnet C et al.; In the absence of recycling, water represents over 90% of the life-support consumables for a manned spacecraft . In addition, over 90% of the waste water generated can be classified as moderately or slightly contaminated (e.g . shower water, condensate from the air-conditioning system, etc.) . The ability to recover potable water from moderately contaminated waste water hence enables significant savings to be made in resupply costs . A development model of such a water-recovery system, based on membrane technology has been produced and tested using "real waste water" based on used shower water Results indicate some 95% recovery of potable water meeting ESA standards, with total elimination of microbial contaminants such as bacteria, spores and viruses.

Proc Natl Acad Sci U S A, 2001 Dec 4, 98(25), 14530 - 5 Epub 2001 Nov 27.
High frequency mitotic gene conversion in genetic hybrids of the oomycete Phytophthora sojae; Chamnanpunt J et al.; Microbial populations depend on genetic variation to respond to novel environmental challenges . Plant pathogens are notorious for their ability to overcome pesticides and host resistance genes as a result of genetic changes . We report here that in particular hybrid strains of Phytophthora sojae, an oomycete pathogen of soybean, high frequency mitotic gene conversion rapidly converts heterozygous loci to homozygosity, resulting in heterokaryons containing highly diverse populations of diploid nuclei . In hybrids involving strain P7076, conversion rates of up to 3 x 10(-2) per locus per nucleus per generation were observed . In other hybrids, rates were of the order of 5 x 10(-5) . Independent gene conversion was observed within a selected linkage group including loci as close as 0.7 kb apart and in unlinked markers throughout the genome . Gene conversions continued throughout vegetative growth and were stimulated by further sexual reproduction . At many loci, conversion showed extreme disparity, with one allele always being lost, suggesting that conversion was initiated by allele-specific double-stranded breaks . Pedigree analysis indicated that individual loci undergo multiple independent conversions within the nuclei of a vegetative clone and that conversion may be preceded by a heritable "activation" state.

Int J Infect Dis, 2001, 5(3), 151 - 5
Rat-bite fever (Streptobacillus moniliformis): a potential emerging disease; Graves MH et al.; OBJECTIVES: To determine the relative prevalence of human infections attributable to Streptobacillus moniliformis in California over the past 3 decades . METHODS: A retrospective analysis of all the data collected was conducted on S . moniliformis cultures identified by the Microbial Diseases Laboratory (MDL) from January 1970 to December 1998 . RESULTS: Information on a total of 45 S . moniliformis isolates was analyzed . Overall, 91% of the isolates were from human sources; 58% were received since 1990 . These strains were divided almost equally between males and females, with 50% of the isolates from patients 9 years old or younger . In 75% of the cases of human infections where a diagnosis was given, rat-bite fever (RBF) was suspected; 83% of these suspected cases involved either a known rat bite or exposure to rodents . CONCLUSIONS: As crowding becomes an increasing environmental reality, humans are more frequently being exposed to zoonotic diseases as a result of encounters with "wild" animals . Domesticated animals also are exposed more frequently to wild animals; thus, increasing human exposure to once rare zoonotic illnesses . Rat-bite fever is a disease that seems to be easily recognizable by clinicians, easily identified in the clinical laboratory (if suspected), and successfully treated when the appropriate therapy is administered . Physicians should consider RBF as a possible diagnosis when fever, rash, and exposure to rats are part of the patient's history.

Water Sci Technol, 2001, 44(7), 165 - 71
Microbial degradation of benzene, toluene, ethylbenzene and xylene isomers (BTEX) contaminated groundwater in Korea; Chang SW et al.; A mixed culture derived from a gasoline-contaminated aquifer in Korea was enriched on toluene at 25 degrees C . A study was conducted to characterize the substrate interaction of BTEX by toluene-enriched consortia and determine the effects of initial BTEX concentration on BTEX degradation . Substrate degradation patterns in individual aromatics were found to differ significantly from patterns for aromatics in mixtures . In the experiment of a single substrate, toluene was degraded fastest, followed by benzene, ethylbenzene, and the xylenes . In BTEX mixtures, degradation followed the order of toluene, ethylbenzene, benzene, and the xylenes . The studies conducting with toluene-enriched consortia evaluated substrate interactions by the concurrent presence of multiple BTEX compounds and revealed a range of substrate interaction patterns including no interaction, stimulation, inhibition, and cometabolism . The simultaneous presence of benzene and toluene were degraded with a slight inhibitory effect on each other . Ethylbenzene was shown to be the most potent inhibitor of BTEX degradation . p-xylene also inhibited the degradation of benzene, toluene, and ethylbenzene, whereas the presence of either benzene or toluene enhanced the degradation of ethylbenzene and the xylenes.

Appl Environ Microbiol, 2001 Dec, 67(12), 5849 - 54
Variation of microbial communities in soil, rhizosphere, and rhizoplane in response to crop species, soil type, and crop development; Wieland G et al.; We investigated the influence of plant species, soil type, and plant development time on the shaping of microbial communities in soil and in association with roots . The sample group consisted of a total of 32 microcosms in three habitats: soil, rhizosphere, and rhizoplane . Communities were represented by the patterns of a sequence-specific separation of rRNA target sequences . Effects of experimental parameters were classified by a cluster analysis of pattern similarities . The type of plant species (clover, bean, or alfalfa) had the greatest effect in plant-associated habitats and also affected soil patterns . Plant development had a minor habitat-dependent effect that was partly obscured by replicate variation . The results stress the applicability of biased community representations in an analysis of induced variation.

Appl Environ Microbiol, 2001 Dec, 67(12), 5780 - 90
Development and evaluation of functional gene arrays for detection of selected genes in the environment; Wu L et al.; To determine the potential of DNA array technology for assessing functional gene diversity and distribution, a prototype microarray was constructed with genes involved in nitrogen cycling: nitrite reductase (nirS and nirK) genes, ammonia mono-oxygenase (amoA) genes, and methane mono-oxygenase (pmoA) genes from pure cultures and those cloned from marine sediments . In experiments using glass slide microarrays, genes possessing less than 80 to 85% sequence identity were differentiated under hybridization conditions of high stringency (65 degrees C) . The detection limit for nirS genes was approximately 1 ng of pure genomic DNA and 25 ng of soil community DNA using our optimized protocol . A linear quantitative relationship (r(2) = 0.89 to 0.94) was observed between signal intensity and target DNA concentration over a range of 1 to 100 ng for genomic DNA (or genomic DNA equivalent) from both pure cultures and mixed communities . However, the quantitative capacity of microarrays for measuring the relative abundance of targeted genes in complex environmental samples is less clear due to divergent target sequences . Sequence divergence and probe length affected hybridization signal intensity within a certain range of sequence identity and size, respectively . This prototype functional gene array did reveal differences in the apparent distribution of nir and amoA and pmoA gene families in sediment and soil samples . Our results indicate that glass-based microarray hybridization has potential as a tool for revealing functional gene composition in natural microbial communities; however, more work is needed to improve sensitivity and quantitation and to understand the associated issue of specificity.

Plant J, 2001 Oct, 28(2), 159 - 68
The mobility of the tobacco Tnt1 retrotransposon correlates with its transcriptional activation by fungal factors; Melayah D et al.; We have analyzed the stress-induced amplification of the tobacco Tnt1 element, one of the rare active plant retrotransposons . Tnt1 mobility was monitored using the retrotransposon-anchored SSAP strategy that allows the screening of multiple insertion sites of high copy number elements . We have screened for Tnt1 insertion polymorphisms in plants regenerated from mesophyll leaf cells, either via explant culture or via protoplast isolation . The second procedure includes an overnight exposure to fungal extracts known to induce high levels of Tnt1 transcription . Newly transposed Tnt1 copies were detected in nearly 25% of the plants regenerated via protoplast isolation, and in less than 3% of the plants derived from explant culture . These results show that Tnt1 transcription is followed by transposition, and that fungal extracts efficiently activate Tnt1 mobility . Transcription appears to be the key step to controlling Tnt1 amplification, as newly transposed Tnt1 copies show high sequence similarities to the subpopulations of transcribed Tnt1 elements . Our results provide direct evidence that factors of microbial origin are able to induce retrotransposon amplification in plants, and strengthen the hypothesis that stress modulation of transposable elements might play a role in generating host genetic plasticity in response to environmental stresses.

Immunology, 2001 Nov, 104(3), 243 - 51
Glycolipid targets of CD1-mediated T-cell responses; Moody DB et al.; Members of the CD1 family of antigen-presenting molecules bind and present a variety of mammalian and microbial glycolipids for specific recognition by T cells . CD1 proteins accomplish their antigen-presenting function by binding the alkyl chains of the antigens within a deep, hydrophobic groove on the membrane distal surface of CD1, making the hydrophilic elements of the antigen available for contact with the variable regions of antigen-specific T-cell receptors . Most models of CD1-restricted T cells function in infectious, neoplastic, or autoimmune diseases and are based on the premise that CD1-restricted T-cell responses are initiated by alterations in cellular glycolipid content . Although a growing number of self, altered self and foreign glycolipid antigens have been identified, the cellular mechanisms that could lead to the generation of antigenic glycolipids within cells, or control the presentation of particular classes of altered self or microbial glycolipids in disease states have only recently come under investigation . Here we review the structures of known glycolipid antigens for T cells and discuss how the chemical nature of these antigens, which is quite different from that of peptides, influences their recognition by T cells.

Environ Microbiol, 2001 Oct, 3(10), 662 - 6
Upstream-independent ribosomal RNA amplification analysis (URA): a new approach to characterizing the diversity of natural microbial communities; Yakimov MM et al.; Here, we propose an advanced method for recently developed fingerprinting strategies to analyse microbial populations by direct detection of 16S rRNA sequences occurring in natural habitats . The differential display (DD) technique, which is widely used to analyse for eukaryotic gene expression, was optimized to assess bacterial rRNA diversity in environmental samples . Double-stranded cDNAs of rRNAs were synthesized without a forward primer digested with endonuclease and ligated with a double-stranded adapter . The fragments obtained were then amplified using an adapter-specific extended primer and a 16S rDNA universal reverse primer pair displayed by electrophoresis on a polyacrylamide gel . We validated this approach by characterization of a microbial community colonizing a geothermal (48 degrees C) vent system located close to the eruption zone of the south-east crater of the Mount Etna volcano, Sicily . Analysis of the patterns of abundant 16S rRNA revealed a considerable diversity of metabolically active bacteria phylogenetically clustering within the Crenarchaeota, Cyanobacteria, Firmicutes, Planctomycetales and Thermus divisions . Two sequence phylotypes were affiliated with uncultivated representatives of the recently described candidate division OP10 from a Yellowstone hot spring.

Waste Manag Res, 2001 Apr, 19(2), 98 - 107
Experimental generation of organic dust from compostable household waste; Heldal KK et al.; The objective was to assess the influence of different waste storage systems on the emission of bioaerosols and gases from compostable household waste . Batches of waste were stored for 14 days in different storage systems: ventilated containers (compostainers) with or without added structure material and closed containers with or without a preservation additive . The microbial potential of the waste was measured with a rotating drum after storage . The weight loss in the compostainers (39%) was higher than in the closed containers (9%) . Hydrogen sulphide and mercaptans developed in the closed container, and the concentration of ammonia increased continuously in both systems to 140 ppm . The microbial content for the incubated waste was high for closed containers compared to compostainers, and waste in closed containers generated a liquid rich in endotoxin and bacteria . The aerosols emitted from the waste consisted mainly of fungal spores, especially Aspergillus fumigatus, and no significant differences were observed between the systems . The endotoxin potential was high for waste stored in closed containers . The use of a preservative prevented microbial growth and reduced the emission of bioaerosols and gases substantially.

J Anim Sci, 2001 Oct, 79(10), 2634 - 42
Effects of level of supplemental phytase on ileal digestibility of amino acids, calcium, and phosphorus in dehulled soybean meal for growing pigs; Traylor SL et al.; Ileally cannulated pigs were used to assess the effects of four dietary levels of microbial phytase (Natuphos) on the apparent and true digestibility of Ca, P, CP, and AA in dehulled soybean meal . Fourteen pigs (25 kg initial BW) were surgically fitted with T-cannulas at the terminal ileum and assigned to diets in a replicated 7 x 7 Latin square design . Following a 14-d recovery, four diets consisting of 30.5% soybean meal with 0, 500, 1,000, or 1,500 units of phytase/kg of diet were fed . Diets 5 (1.05% lysine, 0.90% Ca, and 0.75% P) and 6 (1.05% lysine, 0.90% Ca, and 0.75% P) contained 35.25% soybean meal and 27.0% soy protein concentrate, respectively . Diet 7 (0.37% lysine, 0.03% Ca, and 0.05% P) was a low-CP, casein-based diet used to estimate the nonspecific endogenous losses of Ca, P, CP, and AA in order to estimate the true digestibility of these nutrients . All diets contained cornstarch and dextrose and were fortified with vitamins and minerals . Chromic oxide was used as an indigestible indicator . The diets were fed daily at 9% of metabolic BW (BW0.75) . Apparent and true ileal digestibility of P increased quadratically (P < 0.01) and true digestibility of Ca increased linearly (P < 0.07) with increasing levels of phytase . Apparent digestibility of Ca was unaffected (P = 0.15) by phytase level . Apparent and true ileal digestibility of CP and most AA increased slightly with the addition of 500 units of phytase/kg of diet, but not at higher levels of phytase supplementation (in most cases, cubic effect, P < 0.05) . Apparent and true ileal nutrient digestibility coefficients were unaffected by soybean meal source (Diet 1 vs Diet 5), except for arginine and Ca . The apparent and true digestibility coefficients for most of the AA tended (P < 0.10) to be lower in diets containing soy protein concentrate vs the common source of soybean meal used in Diet 5, but ileal digestibilities of Ca and P were unaffected (P = 0.15) . In this study, supplemental microbial phytase did not improve the utilization of AA provided by soybean meal but was an effective means of improving Ca and P utilization by growing swine fed soybean meal-based diets.

Pest Manag Sci, 2001 Nov, 57(11), 1043 - 54
Quantitative structure-transformation relationships of phenylurea herbicides; Berger BM et al.; Quantitative relationships between the structure of phenylurea herbicides and their transformation in different matrices have been developed . Experimental data on microbial transformation by pure and mixed cultures of soil micro-organisms in inoculated and native soil, as well as on chemical transformation by hydrolysis in sterile soil and water, were available from previous studies . Around 60 experimental or calculated descriptors were used . Quantum chemical calculations were performed with three different semi-empirical methods . Models developed with multiple linear regression were generally easier to interpret than those derived with partial least-squares projection to latent structures . Quite simple and interpretable models could be found to predict transformation rates by pure cultures from lipophilicity, by mixed cultures from adsorption distribution coefficients, and by chemical or enzymatic hydrolysis from electronic properties . Transformation in inoculated soil could not be predicted, but for native soil the use of a quantum chemical descriptor for reactivity (energy of LUMO) together with molar refraction resulted in a general model.

Pest Manag Sci, 2001 Nov, 57(11), 1023 - 32
Rate of bentazone transformation in four layers of a humic sandy soil profile with fluctuating water table; Leistra M et al.; The rate of transformation of a pesticide as a function of the depth in the soil is needed as an input into computations on the risk of residues leaching to groundwater . The herbicide bentazone was incubated at 15 degrees C in soil materials derived from four layers at depths of up to 2.5 m in a humic sandy soil profile with a fluctuating water table (0.8 to 1.4 m), while simulating the redox conditions existing in the field . Gamma-irradiation experiments indicated that bentazone is mainly transformed by microbial activity in the soil . The rate constant for transformation was highest in the humic sandy top layer; it decreased with depth in the sandy vadose subsoil . However, material from the top of the phreatic aquifer had a higher rate constant than that from the layers just above . The presence of fossil organic material in the fluviatile water-saturated sediment probably stimulated microbial activity and bentazone transformation . The changes in the transformation rate constant with depth showed the same trend as those in some soil factors, viz organic carbon content, water-extractable phosphorus and microbial density as measured by fluorescence counts . However, the (low) concentration of dissolved organic carbon (DOC) in the top of the aquifer did not fit the trend . The rate constant for bentazone transformation in the layers was higher at lower initial contents of the herbicide.

Waste Manag Res, 2001 Aug, 19(4), 284 - 91
Stability evaluation of a cement based waste form to microbially induced degradation; Idachaba MA et al.; In this study the current Nuclear Regulatory Commission (NRC) protocol is used to evaluate the stability of Tuskegee cement/cobalt chloride waste form in the presence of Thiobacillus thiooxidans (T . thiooxidans) . A critical examination of this protocol and identified limitations are reported also . Tuskegee cement/cobalt chloride waste forms were shown to exhibit considerable instability to microbial degradation as indicated by significant physical deterioration, and increased leaching of calcium and cobalt on exposure to T . thiooxidans . The instability was aggravated with higher levels of cobalt chloride content of the waste forms . The degradative capability of T . thiooxidans closely followed its ability to significantly decrease the pH of its environment . Inherent limitations in the NRC protocol were observed which could lead to serious result interpretation errors . The use of a T . thiooxidans culture that is significantly lower in pH in comparison to the control medium could lead to an overestimation of the degradative effect of T . thiooxidans, while the use of a culture that is substrate limited could result in an underestimation of T . thiooxidans capability.

Brain Res Bull, 2001 Oct-Nov 1, 56(3-4), 281 - 4
The channel hypothesis of Huntington's disease; Kagan BL et al.; Extended tracts of polyglutamine (PG) have been implicated in the pathogenicity of the mutant protein huntingtin and have been shown to form ion channels in planar lipid bilayers . These lines of evidence suggest that huntingtin and other PG mutant proteins may damage cells via a channel mechanism . This mechanism could cause damage to the plasma membrane by running down ionic gradients, discharging membrane potential; or allowing influx of toxic ions such as Ca(2+) . PG damage to intracellular membranes such as the lysosomal membrane or the mitochondrial membrane could also injure cells via leakage of toxic enzymes or triggering of apoptosis . The channel mechanism is well-established for microbial toxins, and the existence of at least six other "amyloid" channels relevant to diseases such as Alzheimer's and Creutzfeld-Jakob, suggests that this may be a widespread pathogenic mechanism.

Mutat Res, 2002 Jan 15, 513(1-2), 143 - 50
Evaluation of the genotoxic potential of some microbial volatile organic compounds (MVOC) with the comet assay, the micronucleus assay and the HPRT gene mutation assay; Kreja L et al.; Microbial volatile organic compounds (MVOC), metabolites of fungi detected in indoor moulds and in working places in compost facilities are considered as a potential health hazard . Their toxicological relevance, however, is largely unknown and data are rare . The aim of this study was to evaluate in vitro the genotoxic, clastogenic and mutagenic potential of same typical MVOC . For the study of DNA damage human lung carcinoma epithelial A549 cells, V79 Chinese hamster fibroblasts and human peripheral blood cells were exposed and subjected to the alkaline comet assay (single cell gel test) . Taking the Chinese hamster V79 cell line as a target clastogenic effects were studied by the micronucleus test and mutagenic effects by the hypoxanthine-guanine-phosphoribosyl transferase gene mutation test (HPRT test) . The cytogenic effects of MVOC were assessed by a clonogenic assay using the A549 cell line . The alkylating agent methyl methanesulfonate (MMS) was taken as a positive control . The results indicate that MVOC induced DNA damage is only seen in conditions in which also cytotoxic effects are observed . Clastogenic and mutagenic effects could not be detected.

New Microbiol, 2001 Oct, 24(4), 389 - 96
Adherence of Actinobacillus actinomycetemcomitans on oral epithelial cells; Gasparetto A et al.; Actinobacillus actinomycetemcomitans is capable of colonizing mucosa and dental plaque and plays an important role in periodontal disease in young peoples and adult . Adherence mechanisms on epithelial cells, tooth or oral bacteria and gingival invasion probably are the initial steps in the pathogenesis of gingivitis or periodontitis . In this study, the adherence of A . actinomycetemcomitans on oral epithelial cells following subculturing were examined . The adherence on oral epithelial cells showed high in all the isolates values but with differences among them and at each time of subculturing . The adherence of A . actinomycetemcomitans FDC Y4 was stable in each of the subcultures . However, adhesion values of all the tested isolates were different except for strains #1, #38 and Y4, suggesting a heterogenicity within this microbial group . Morphologic variations were observed in extracellular structures of the A . actinomycetemcomitans tested . The adhesion process on oral epithelial cells of this organism can be influenced by subcultures, but additional studies are necessary to verify the influence of subculturing on adherence or other virulence factors.

Environ Sci Technol, 2001 Nov 1, 35(21), 4341 - 6
Hexahydro-1,3,5-trinitro-1,3,5-triazine mineralization by zerovalent iron and mixed anaerobic cultures; Oh BT et al.; Soil microcosms were used to evaluate the potential benefits of an integrated microbial-Fe0 system to treat groundwater contamination by RDX (hexahydro-1,3,5-trinitro-1,3,5-triazine) . Microcosms amended with both Fe0 filings and municipal anaerobic sludge mineralized RDX faster and to a greater extent than separate treatments, with up to 51% 14CO2 recovery after 77 d . The nitroso byproducts 1,3-dinitro-5-nitroso-1,3,5-triazacyclohexane (MNX), 1,3-dinitroso-5-nitro-1,3,5-triazacyclohexane (DNX), and 1,3,5-trinitroso-1,3,5-triazacyclohexane (TNX) were detected in all microcosms, although these compounds never accumulated above 5% of the added RDX on a molar basis . A soluble intermediate that was tentatively identified as methylenedinitramine {(O2NNH)2CH2} was relatively persistent, although it accumulated to a much lower extent in combined-treatment reactors than in sets with Fe0 or sludge alone . Some of the radiolabel was bound to soil and Fe0 and could not be extracted with CH3CN . This fraction, which was recovered by combustion with a biological oxidizer, was also found at lower concentrations in combined-treatment reactors . This work suggests that permeable reactive Fe0 barriers might be an effective approach to intercept and degrade RDX plumes and that treatment efficiency might be enhanced by biogeochemical interactions through bioaugmentation.

Environ Sci Technol, 2001 Nov 1, 35(21), 4265 - 70
Study of the degradation of the herbicides 2,4-D and MCPA at different depths in contaminated agricultural soil; Crespin MA et al.; Two phenoxyacid herbicides (2,4-D and MCPA) and their six corresponding phenols were determined in soil by using gas chomatography with electron impact mass spectrometry (GC/MS) for confirmation/quantitation . An automatic extraction (leaching), preconcentration, and cleanup (sorption) module was developed to extract the eight compounds from soil . The average recovery of all species, spiked to soil at microg/kg-mg/kg levels, was 95% (average standard deviation +/- 5%) . A plot of agricultural clayey soil (approximately 12 m2) was contaminated with both herbicides (approximately 96 g/m3, depth 10 cm, density 1.23 g/cm3) and irrigated with (17 mm) at variable time intervals . Both herbicides and their corresponding phenol compounds were monitored at different soil depths over a 50 day period . The degradation of both herbicides in the surface layer (t(1/2) approximately 5 days) is a result of photodecomposition and microbial action; in the deeper layers, the degradation products occur in lower proportions by effect of leaching and are also the result of microbial action . The six phenol metabolites are only detected in the surface layer as they form preferentially by photodecomposition . The main metabolites (viz . 2,4-DCP for 2,4-D and 4-C-2-MP for MCPA) are formed within 24 h after the soil is contaminated; their concentration peaks are at day 8 in the absence of irrigation.

J Endotoxin Res, 2001, 7(4), 327 - 32
The crucial role of systemic responses in the innate (non-adaptive) host defense; Munford RS et al.; We suggest that successful defense against microbial invasion requires both local inflammation and systemic anti-inflammation . The key systemic responses involve the hypothalamic-pituitary-adrenocortical axis, the sympathetic-adrenomedullary axis, acute phase protein production, thermoregulation and alterations in leukocyte responsiveness to agonists such as bacterial endotoxin . These integrated responses raise blood and tissue concentrations of several anti-infective molecules, mobilize leukocytes into the circulation, and increase blood flow to injured or infected sites . They also neutralize cytokines, proteases and oxidants that enter the bloodstream from inflamed local sites and forestall endothelial activation in distant vessels . Together, these forces help concentrate activated phagocytes at injured or infected local sites while preventing potentially damaging inflammation in uninvolved tissues.

Eur Respir J, 2001 Oct, 18(4), 692 - 704
Lung dendritic cells and host immunity to infection; Lambrecht BN et al.; The lung is a portal of entry for numerous microbial pathogens, against which evolution has created an adequate innate and adaptive immune response . Dendritic cells (DCs) are central to the integration of innate and specific immunity . These cells are located within the epithelium and interstitium of the lung where they are influenced by the innate immune system . Upon recognition and internalization of microbial antigens, DCs migrate to the draining lymph nodes of the lung to initiate the specific cellular and humoral immune response . By their capacity to integrate stimuli derived from the pathogen, the host and the environment, they are specialized to induce a protective immune response while at the same time avoiding damage to the host . It is becoming increasingly clear that dendritic cells are involved in the induction of immunity to viruses, bacteria, mycobacteria and fungi . Some pathogens subvert the function of dendritic cells to escape immune recognition . Not surprisingly, if dendritic cell function fails, the consequence for the host is immunodeficiency.

Nahrung, 2001 Oct, 45(5), 324 - 31
Enzymatic degradation studies of pectin and cellulose from red beets; Dongowski G; The influence of structural features of the cell wall polysaccharides pectin and cellulose on the enzymatic degradation of red beet was evaluated . Alcohol-insoluble substances and acetone-insoluble residues were prepared from red beets and characterized with respect to the content of dietary fibre, pectin fractions, neutral saccharide composition and water absorption . The high-methylated and high-acetylated pectin component was partly soluble in water and in EDTA . Pectin was hardly extractable from alcohol-insoluble substances as well as from red beets . Isolated pectin could not be completely degraded by pectolytic enzymes . After de-acetylation, the pectic acid from red beets was degradable in a similar rate like citrus pectic acid . From alcohol-insoluble substances, several cellulose and lignin fractions were prepared and analysed . A cellulose preparation from red beets was intensely degraded by cellulases with high activities as shown by the release of reducing end-groups, viscosity and scanning electron microscopy . Cell wall preparations from red beets were able to bind high amounts of water . A decrease in water absorption during enzymatic action or changes in viscosity and flow behaviour are sensitive markers for decomposition or depolymerization processes . Furthermore, an inhibitor of microbial enzymes was isolated from red beets and acetone-insoluble residues . The main reason for the poor enzymatic liquefaction or maceration of red beets by pectolytic and cellulolytic enzymes is the high degree of acetylation of the pectin component.

J Immunol, 2001 Dec 1, 167(11), 6503 - 9
CXCR2 deficiency confers impaired neutrophil recruitment and increased susceptibility during Toxoplasma gondii infection; Del Rio L et al.; Neutrophil migration to the site of infection is a critical early step in host immunity to microbial pathogens, in which chemokines and their receptors play an important role . In this work, mice deficient in expression of the chemokine receptor CXCR2 were infected with Toxoplasma gondii and the outcome was monitored . Gene-deleted animals displayed completely defective neutrophil recruitment, which was apparent at 4 h and sustained for at least 36 h . Kit(W)/Kit(W-v) animals also displayed defective polymorphonuclear leukocyte migration, suggesting mast cells as one source of chemokines driving the response . Tachyzoite infection and replication were accelerated in CXCR2(-/-) animals, resulting in establishment of higher cyst numbers in the brain relative to wild-type controls . Furthermore, serum and spleen cell IFN-gamma levels in infected, gene-deleted mice were reduced 60-75% relative to infected normal animals, and spleen cell TNF-alpha was likewise reduced by approximately 50% . These results highlight an important role for CXCR2 in neutrophil migration, which may be important for early control of infection and induction of immunity during Toxoplasma infection.

J Immunol, 2001 Dec 1, 167(11), 6421 - 30
Functional and structural similarity of V gamma 9V delta 2 T cells in humans and Aotus monkeys, a primate infection model for Plasmodium falciparum malaria; Daubenberger CA et al.; Gammadelta T cells are implicated to play crucial roles during early immune responses to pathogens . A subset of human gammadelta T cells carrying the Vgamma9Vdelta2 TCR recognize small, phosphorylated nonpeptidic Ags . However, the precise role of these cells and the ligands recognized in human immune responses against pathogens remains unclear because of the lack of suitable animal models . We have analyzed the reactivity of spleen cells of the New World monkey Aotus nancymaae against isopentenyl pyrophosphate (IPP), a phosphorylated microbial metabolite selectively activating Vgamma9Vdelta2 T cells . Spleen cells were stimulated by IPP and the expanding cell population expressed the Vgamma9 TCR . TRGV-J and TRDV-D-J rearrangements expressed by IPP-stimulated cells of Aotus were analyzed by RT-PCR and DNA sequencing . The TRGV-J and TRDV-D-J rearrangements expressed by IPP-stimulated Aotus and human gammadelta T cells were similar with respect to 1) TCR gene segment usage, 2) a high degree of germline sequence homology of the TCR gene segments used, and 3) the diversity of the CDR3 regions . Phylogenetic analysis of human, Pan troglodytes, and A . nancymaae TRGV gene segments showed that the interspecies differences are smaller than the intraspecies differences with TRGV9 gene segments located on a distinct clade of the phylogenetic tree . The structural and functional conservation of Vgamma9Vdelta2 T cells in A . nancymaae and humans implicates a functionally important and evolutionary conserved mechanism of recognition of phosphorylated microbial metabolites.

J Immunol, 2001 Dec 1, 167(11), 6247 - 55
Lipopolysaccharide modulation of dendritic cells is insufficient to mature dendritic cells to generate CTLs from naive polyclonal CD8+ T cells in vitro, whereas CD40 ligation is essential; Kelleher M et al.; Many cytotoxic CD8+ T cell responses are dependent on the interactions between CD40 ligand on the helper CD4+ T cell and CD40 on the APC . Although CD40 triggering of dendritic cells (DC) has been shown to mature the DC by increasing the level of expression of costimulatory molecules and inducing IL-12 secretion, the precise mechanisms by which CD40-CD40 ligand interactions allow DC to drive CTL responses remain unknown . We have used an in vitro model in which naive polyclonal CD8+ T cells can be activated by bone marrow-derived DC to investigate factor(s) that are responsible for this CD40-dependent generation of CTLs . DC modulated with agonistic anti-CD40 mAb (aCD40) are able to generate Ag-specific CTL responses while DC modulated with the microbial stimulus LPS alone do not . We compared the Ag-presenting capacity, levels of costimulatory molecules, and release of cytokines and chemokines of DC modulated with aCD40 to that of DC modulated by LPS . None of the factors assayed account for the unique capacity of anti-CD40-matured DC to drive CTL but this model provides a simplified system for further investigation . Although we attempted to use an LPS-free system for these studies, we are unable to rule out the possibility that very low levels of endotoxin (<20 pg/ml) may synergize with CD40 ligation in the generation of CTLs.

Acc Chem Res, 2001 Nov, 34(11), 919 - 30
Designer hybrid cyclopeptides for membrane ion transport and tubular structures; Ranganathan D; The incorporation of chosen, non-amino acids into cyclopeptides is a promising approach to designs capable of performing specific tasks . While such hybrid cyclopeptides are common in the microbial world, there is hardly any effort by synthetic organic chemists to explore such designs for creating architecturally beautiful and functionally useful macrocyclic hybrid peptides . This Account, mainly reviewing the author's own work, presents a simple design strategy enabling the crafting of a large variety of hybrid cyclopeptides and leading to the identification of excellent membrane ion carriers, units that self-assemble into tubular structures, and systems for specific guest recognition.

Am J Respir Cell Mol Biol, 2001 Nov, 25(5), 606 - 12
What is the role of nitric oxide in murine and human host defense against tuberculosis?Current knowledge; Chan ED et al.; The production of reactive oxygen intermediates and reactive nitrogen intermediates by innate immune cells is considered to be an effective host-defense mechanism against microbial pathogens . In the murine model of tuberculosis (TB), nitric oxide (NO) plays an essential role in the killing of Mycobacterium tuberculosis by mononuclear phagocytes . For example, in the mouse strain with a genetic disruption for inducible NO synthase (iNOS-/-), infection with M . tuberculosis is associated with a significantly higher risk of dissemination and mortality . Although more controversial in humans, there is a growing body of evidence that NO produced by TB-infected macrophages and by epithelial cells also has antimycobacterial effects against M . tuberculosis . The precise mechanism(s) by which NO and other reactive nitrogen species antagonize M . tuberculosis is not known, but may involve disruption of bacterial DNA, proteins, signaling, and/or induction of apoptosis of macrophages that harbor mycobacteria . In addition to cytokines such as tumor necrosis factor-alpha and interleukin 1-beta, mycobacterial cell wall components such as lipoarabinomannan and 19 kD lipoprotein, along with the T-cell-derived interferon-gamma, may also induce NO expression . In a Darwinian fashion, it also appears that certain strains of M . tuberculosis have evolved strategies to combat the toxic effects of NO.

Braz J Infect Dis, 2001 Aug, 5(4), 223 - 32
Successful use of a defined antigen/GM-CSF adjuvant vaccine to treat mucosal Leishmaniasis refractory to antimony: A case report; Badaro R et al.; Immunotherapy has been proposed as a method to treat mucosal leishmaniasis for many years, but the approach has been hampered by poor definition and variability of antigens used, and results have been inconclusive . We report here a case of antimonial-refractory mucosal leishmaniasis in a 45 year old male who was treated with three single injections (one per month) with a cocktail of four Leishmania recombinant antigens selected after documented hypo-responsiveness of the patient to these antigens, plus 50 microg of GM-CSF as vaccine adjuvant . Three months after treatment, all lesions had resolved completely and the patient remains without relapse after two years . Side effects of the treatment included only moderate erythema and induration at the injection site after the second and third injections . We conclude that carefully selected microbial antigens and cytokine adjuvant can be successful as immunotherapy for patients with antimonial-refractory mucosal leishmaniasis.

J Invest Dermatol, 2001 Nov, 117(5), 1193 - 9
Ultraviolet a radiation suppresses an established immune response: implications for sunscreen design; Nghiem DX et al.; The ultraviolet radiation present in sunlight is the primary cause of nonmelanoma skin cancer and has been implicated in the development of cutaneous malignant melanoma . In addition, ultraviolet is immune suppressive and the suppression induced by ultraviolet radiation has been identified as a risk factor for skin cancer induction . Ultraviolet also suppresses the immune response to infectious agents . In most experimental models, ultraviolet is applied to immunologically naive animals prior to immunization . Of equal concern, however, is the ability of sunlight to suppress established immune reactions, such as the recall reaction in humans, which protects against microbial infections . Here we demonstrate that solar-simulated ultraviolet radiation, applied after immunization, suppresses immunologic memory and the elicitation of delayed-type hypersensitivity . Further, we found that wavelengths in the ultraviolet A region of the solar spectrum were critical for inducing immune suppression . Ultraviolet A (320-400 nm) radiation was as effective as solar-simulated ultraviolet A + B (290-400 nm) in suppressing the elicitation of an established immune response . Irradiation with ultraviolet AI (340-400 nm) had no effect . Supporting a critical role for ultraviolet A in ultraviolet-induced immune suppression was the observation that applying a sunscreen that contained an ultraviolet B only filter had no protective effect, whereas, a sunscreen containing both ultraviolet A and ultraviolet B filters totally blocked ultraviolet-induced immune suppression . These data suggest that sunlight may depress the protective effect of prior vaccination . In addition, the observation that ultraviolet A is immunosuppressive indicates the need for ultraviolet A protection when designing sun protection strategies.

Int Immunopharmacol, 2001 Nov, 1(12), 2043 - 62
Factors modifying the migration of lymphocytes across the blood-brain barrier; Brown KA; Characterising the factors that control the entry of leucocytes into tissue in response to inflammatory or microbial insult continues to generate considerable interest . Of all the tissues studied it is probably that of the CNS which is the most fascinating because of the specialised properties of its blood vessel walls, which constitute the blood-brain barrier (BBB) . In health, very few leucocytes penetrate the BBB but in disorders such as MS the barrier becomes compromised with the result that there is an intense infiltration of the CNS by T lymphocytes whose subsequent activity appears to underlie the onset and progression of disease . The purpose of this article is to summarise and assess recent literature pertaining to how lymphocytes bind to cerebral endothelial cells, migrate across the blood vessel walls and enter the CNS parenchyma . Particular emphasis is devoted to the cellular and molecular aspects of these events and addressing the questions of whether certain subsets of circulating T lymphocytes are more favourably disposed than others to CNS infiltration and whether entry is dependent upon the initial expression of distinct groups of adhesion molecules and upon the generation of chemotactic factors . This article also focuses upon identifying the key stages of lymphocyte migration across the BBB and their susceptibility to antagonism by therapeutic agents . It is intended that the review will provide a useful source of information and offer additional insights into the mechanisms controlling lymphocyte passage across the BBB during pathological disturbance.

Biomacromolecules, 2000 Summer, 1(2), 157 - 61
Microbial degradation of poly(aspartic acid) by two isolated strains of Pedobacter sp . and Sphingomonas sp; Tabata K et al.; Microbial degradation of thermally synthesized poly(aspartic acid) (PAA) was investigated . A PAA-P1 sample (Mn, 7500; Mw, 20,000; number of branched units/100 monomer units, 3.1) was completely degraded in natural river water within 15 days at 25 degrees C . A new PAA-degrading bacterium (strain KP-2: JCM10638) was isolated together with Sphingomonas sp . KT-1 (JCM10459) from river water, and identified as a member of Pedobacter . A Pedobacter isolate was capable of degrading high-molecular-weight PAA polymers of 5000 to 150,000, and a small amount of low-molecular-weight products of 250 to 5000 was accumulated as residues during the growth of the isolate on PAA . In contrast, the other isolate Sphingomonas sp . KT-1 degraded only low-molecular-weight PAA below 5000 . A mixed culturing of Pedobacter sp . KP-2 with Sphingomonas sp . KT-1 resulted in a complete degradation of PAA-P1 sample, but a small amount of low molecular weight components was accumulated during the degradation of highly branched PAA-P2 and PAA-P3 samples.

Biomacromolecules, 2001 Fall, 2(3), 958 - 64
Enzymatic epimerization of bacterial mannuronan and of C-6 oxidized, galactose-depleted guar: a circular dichroism and 1H NMR study; Crescenzi V et al.; Attention has been focused on two uronans, namely, mannuronan and galactose-depleted C-6 oxidized guar, the former of microbial origin and the latter of artificial nature, to provide original data on the extent of epimerization they can undergo in dilute aqueous solution using two C-5 mannuronic acid epimerizing enzymes, that is, AlgE-4 and AlgE-6, alone or in admixture . Original circular dichroism data coupled with 1H NMR spectra clearly point out that both uronans can be epimerized, depending on the enzyme or enzyme mixture used, to high levels yielding guluronic-rich alginate samples and guluronic-rich heteropolysaccharides, respectively . Mannuronan and its epimerization products can easily form clear, firm aqueous gels when an excess of HCl is added or when mixed with aqueous CaCl2, respectively . Depleted-guarox does not gel upon addition of excess HCl, while the heterouronan derived from it having a percent of epimerization nearly identical to that of epimerized mannuronan, that is, ca . 70%, can form gel in the presence of Ca(II) only at higher polymer and Ca(II) concentrations . With the latter, heterouronan alpha-D-galacturonic side groups exert hindrance to "junction zone" formation.

Biomacromolecules, 2001 Fall, 2(3), 1055 - 60
Synthesis of microbial poly(beta-hydroxybutyrate) modified with oligo(pentaerythritol ethoxylate) by Ralstonia eutropha; Jenzsch M et al.; Poly(beta-hydroxybutyrate) (PHB) modified with different amounts of pentaerythritol ethoxylate (PEE) has been synthesized using Ralstonia eutropha . The growth kinetics and the synthesis of PHB in the presence of PEE were modeled using appropriate differential equations for the mass balance of the two-stage process . The influence of PEE addition on the morphology of PHB was studied by various microscopic and scattering techniques . Light microscopic and wide-angle X-ray measurements indicated that the addition of PEE had a nucleating effect on the crystallization of PHB . The spherulite growth rate was widely independent of the PEE addition . The lamellae of PHB became more disordered when PEE was added as demonstrated by atomic force microscopy . Furthermore, small-angle X-ray data indicated a decrease in the long period with increasing PEE content of the modified PHB.

Ann R Australas Coll Dent Surg, 2000 Oct, 15, 42 - 50
Aetiology and pathogenesis of periodontal disease; Kinane DF; Gingivitis is modified by several factors including smoking, ion channel blocking drugs and hormonal changes such as those seen in puberty or pregnancy . Periodontitis is also initiated by microbial plaque which follows on from gingivitis, but occurs in only 10 to 15% of the population and is influenced by the individual's immune and inflammatory response . Studies on the humoral and cellular immune response in periodontal disease indicate that: a) homing of relevant immune cells takes place within the periodontium; b) plasma cells are among the most active secretory cells in the gingiva; c) that immunoglobulin subclasses are similar between blood and the gingival pocket fluid for IgG but not for IgA; and that d) an individual's ability to mount a specific antibody response to periodontopathogenic organisms may indicate their susceptibility to the disease and their likely response to treatment . Recent reviews of the pathology of periodontal disease suggest that a histopathological established lesion with plasma cell predominance is likely to undergo periodontal bone loss . Furthermore, susceptibility to periodontitis may be related to whether plasma cells predominate in the tissues of an individual or a site as result of the microbial challenge of dental plaque . A tendency for an individual or site to form an extensive plasma cell infiltrate may indicate an inability to defend against periodontal bacteria and thus a predisposition to periodontitis . The various susceptibility factors such as smoking, medical condition, genetics, plaque control and local factors may all interact to influence the host response and specifically the immune response.

Ann R Australas Coll Dent Surg, 2000 Oct, 15, 34 - 41
Periodontal diagnostics; Kinane DF; Periodontitis affects a subset of the population and our current thinking is that progression of periodontal disease may be either continuous or cyclical (burst hypothesis) . These features make screening and diagnostic tools desirable in the management of this disease . Although many potential markers exist, several difficulties hamper our ability to declare them diagnostic tests with proven utility . The 'gold standard' for active periodontal disease is not available and inflammation due to gingivally confined lesions (gingivitis) and periodontal inflammation which results in attachment loss is a potential confounder of any test based on assessing the host response elements of the disease . The current absence of proof for the progression of periodontal disease i.e., whether or not the burst hypothesis is correct, is a further problem . Although much is written about the need for markers of current or future disease which will prevent us from overtreating pockets, the time, effort and cost involved in testing these sites has to be balanced against the relative ease and speed of routine therapy such as root planing . In addition, we are still some way from the development and validation of reliable host or microbial testing methods . In terms of screening tests for diseases such as the early-onset forms of periodontitis, the research and development on diagnostic tools involving genetic polymorphisms, specific genes, systemic antibodies or leucocyte cell surface markers of the patients, may become a clinical reality in time . One could envisage chairside tests using blood from thumb pricks being capable of determining a young individual's risk of developing disease at a later age, and thus the need for a timely prevention programme . Before applying any test we should reconsider what treatment planning effects a positive or negative result will have, and any test which does not influence the treatment plan is redundant . Whether periodontal diagnostic tests will be developed which will indicate to the clinician the need for various treatment regimes other than root planing, is still an open question . For example, in the future, a test of specific antibody levels in patients prior to treatment might indicate their likely response to therapy and thus the need for additional therapeutic agents such as antibiotics . Periodontal diagnostic tests are still at an early stage of development and much work remains to be performed to fully validate their utility such that they become an important and cost effective aspect of clinical treatment planning, screening or patient monitoring.

Ann R Australas Coll Dent Surg, 2000 Oct, 15, 235 - 9
Intraradicular space: what happens within roots of infected teeth?
Love RM.
The pulpo-dentine complex is normally protected from exogenous substances in the oral cavity by the overlying enamel or cementum . Dental caries, dental trauma, enamel/dentine cracks, and restorative procedures commonly breach the integrity of enamel or cementum and may allow infection of the pulpo-dentine complex to occur, possibly leading to pulp and periapical inflammatory disease . Infection of the intraradicular space is a complex and dynamic process involving interactions between host and microbial factors . An understanding of these factors has led to the development of endodontic techniques that offer predictable success.

Curr Issues Intest Microbiol, 2001 Mar, 2(1), 17 - 25
Terminal restriction fragment patterns: a tool for comparing microbial communities and assessing community dynamics; Kitts CL; Terminal Restriction Fragment (TRF) patterns, also known as Terminal Restriction Fragment Length Polymorphisms (T-RFLP), are a recently introduced PCR-based tool for studying microbial community structure and dynamics . Since the first review of TRF methodology (Marsh, 1999 . Curr . Op . Microbiol . 2: 323-7), at least 35 new research articles were published that include this powerful tool in some part of their reports . This review covers some of the applications that TRF patterns were used for and provides a discussion of how to create and analyze TRF pattern data . This data has the advantage of being simply and rapidly produced using standard DNA sequencing equipment . The raw data are automatically converted to a digitized form that can be easily analyzed with a variety of multivariate statistical techniques . The identification of specific elements in a TRF pattern is possible by comparison to entries in a good sequence database or by comparison to a clone library . As an added advantage when investigating complex microbial communities such as those in soils and intestines, TRF patterns are recognized as having better resolution than other DNA-based methods for evaluating community structure.

Curr Issues Intest Microbiol, 2001 Mar, 2(1), 1 - 15
Microbial biota of the human intestine: a tribute to some pioneering scientists; Savage DC; Research on the indigenous intestinal microbiota of man was initiated well before the end of the 19th Century . The work continued at a slow but steady pace throughout the first half of the 20th Century . Findings from the effort had little impact on medicine and other aspects of human biology, however, until the 6th decade of the 20th Century . During that decade, research in the area was begun by eight groups of investigators, each of which was led by one or two senior scientists with great experimental talent, creativity and foresight . Their findings added new dimension to knowledge of the microbiota and initiated an explosion of interest in research in the field that has continued to the present day . The research of the groups during the 1960's is described in this review as a tribute to the senior scientists who had such critical impact on this important field of study.

Microbes Infect, 2001 Nov, 3(13), 1167 - 71
How the immune system protects the host from infection; Janeway CA Jr; The immune system is made up of sets of interacting cells . The first to respond in all cases are the antigen-presenting cells (APCs), which are equipped with receptors for microbial patterns . Engagement of these receptors induces co-stimulatory molecules on the surface of the APCs, and allows it to stimulate potent CD4 T-cell responses, and also CD8 T-cell responses . This in turn leads to B-cell-derived antibody responses . The entire response is controlled by suppressor T cells, as predicted many years ago by Richard Gershon.

Vet Parasitol, 2001 Nov 22, 101(3-4), 187 - 200
Defining and interpreting intraspecific molecular variation; Rosenthal BM; Defining the extent and character of intraspecific genetic variation provides important information about gene function and organismal history . Powerful tests may be applied to sequenced alleles in order to critically examine whether natural selection is responsible for limiting or elevating intraspecific polymorphism in particular genes . Unconventional patterns of sequence variation and unusual allelic frequency distributions can be used to test whether genes encoding parasite antigens are being diversified by immune selection . The strikingly limited genetic variation in the falciparum malaria genome, and in human chromosomes encoding resistance to severe malaria, date the emergence of this disease to within the last few thousand years, illustrating the power of population genetic analysis to elucidate the history of host-parasite interactions . Coupling phylogenetic and geographic information and analyzing the rate of diversification in intraspecific gene trees provides new and rich sources of information on microbial evolution and epidemiology.

Genomics, 2001 Nov, 78(1-2), 12 - 4
Identification, genomic organization, and mRNA expression of LACTB, encoding a serine beta-lactamase-like protein with an amino-terminal transmembrane domain; Smith TS et al.; Database searching with bacterial serine beta-lactamases identified mouse expressed sequence tags (ESTs) with significant similarity scores.The cloned mouse cDNA encodes a novel 551-amino-acid protein, LACTB, with a predicted amino-terminal transmembrane domain but no signal peptide . It contains an active site motif related to C-class beta-lactamases . Homologues were detected in sequence data from human, rat, cow, rabbit, pig, toad, zebrafish, and Caenorhabditis elegans, but not in Saccharomyces cerevisiae or Drosophila melanogaster . The genes were mapped to human chromosome 15q22.1 and mouse chromosome 9 . Sequencing of a 14.7-kb fragment of mouse genomic DNA defined six exons . A virtual human cDNA and a 549-residue protein, predicted from unfinished genomic sequence, showed the same intron/exon structure . Northern blot analysis showed expression of the 2.3-kb mRNA predominantly in mouse liver and human skeletal muscle . This is the first reported vertebrate example of this microbial peptidase family.

Environ Pollut, 2001, 115(2), 219 - 30
A biological loss of endosulfan and related chlorinated organic compounds from aqueous systems in the presence and absence of oxygen; Guerin TF; Endosulfan is a cyclodiene organochlorine currently widely used as an insecticide throughout the world . This study reports that the endosulfan isomers can be readily dissipated from aqueous systems at neutral pH in the absence of biological material or chemical catalysts, in the presence or absence of oxygen . The study showed that aldrin, dieldrin, and endosulfan exhibit bi-phasic loss from water in unsealed and butyl rubber sealed vessels . Half-lives are substantially increased for endosulfan I when oxygen is removed from the incubation vessel . The study conditions, where PTFE was used, were such that loss due to volatilization and alkaline chemical hydrolysis was eliminated . Half-lives determined from these data indicate that the parent isomers are much less persistent than the related cyclodienes, aldrin and dieldrin, confirming the findings of previous studies . The major oxidation product of endosulfans I and II, endosulfan sulfate, is less volatile and can persist longer than either of the parent isomers . Endosulfan sulfate was not formed in any of the treatments suggesting that it would not be formed in aerated waters in the absence of microbial activity or strong chemical oxidants . Since endosulfan sulfate is formed in many environments through biological oxidation, and is only slowly degraded (both chemically in sterile media and biologically), it represents a predominant residue of technical grade endosulfan, which finds its way into aerobic and anaerobic aquatic environments . The data obtained contributes to and confirms the existing body of half-life data on endosulfan I and II and its major oxidation product, endosulfan sulfate . The half-life data generated from the current study can be used in models for predicting the loss of chlorinated cyclodiene compounds from aqueous systems . The findings also highlight the importance of critically reviewing half-life data, to determine what the predominant processes are that are acting on the compounds under study.

Infect Immun, 2001 Dec, 69(12), 7703 - 10
Host response to infection: the role of CpG DNA in induction of cyclooxygenase 2 and nitric oxide synthase 2 in murine macrophages; Ghosh DK et al.; Depending on sequence, bacterial and synthetic DNAs can activate the host immune system and influence the host response to infection . The purpose of this study was to determine the abilities of various phosphorothioate oligonucleotides with cytosine-guanosine-containing motifs (CpG DNA) to activate macrophages to produce nitric oxide (NO) and prostaglandin E(2) (PGE(2)) and to induce expression of NO synthase 2 (NOS2) and cyclooxygenase 2 (COX2) . As little as 0.3 microg of CpG DNA/ml increased NO and PGE(2) production in a dose- and time-dependent fashion in cells of the mouse macrophage cell line J774 . NO and PGE(2) production was noted by 4 to 8 h after initiation of cultures with the CpG DNA, with the kinetics of NO production induced by CpG DNA being comparable to that induced by a combination of lipopolysaccharide and gamma interferon . CpG DNA-treated J774 cells showed enhanced expression of NOS2 and COX2 proteins as determined by immunoblotting, with the relative potencies of the CpG DNAs generally corresponding to those noted for the induction of NO and PGE(2) production as well as to those noted for the induction of interleukin-6 (IL-6), IL-12, and tumor necrosis factor . Extracts from CpG DNA-treated cells converted L-arginine to L-citrulline, but the NOS inhibitor N(G)-monomethyl-L-arginine (NMMA) inhibited this reaction . The COX2-specific inhibitor NS398 inhibited CpG DNA-induced PGE(2) production and inhibited NO production to various degrees . The NOS inhibitors NMMA, 1400W, and N-iminoethyl-L-lysine effectively blocked NO production and increased the production of PGE(2) in a dose-dependent fashion . Thus, analogues of microbial DNA (i.e., CpG DNA) activate mouse macrophage lineage cells for the expression of NOS2 and COX2, with the production of NO and that of PGE(2) occurring in an interdependent manner.

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