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