Environ Res, 2000 Mar, 82(3), 253 - 7 A comparison of sampling media for environmental viable fungi collected in a hospital environment; Wu PC et al.; Quantitative evaluation of fungal exposure is often conducted by analysis of the composition of microbes in air samples and calculation of the concentrations afterward . The collecting medium that favors the growth for most saprophytic fungi is considered to be the ideal choice in most circumstances . Currently, the culture medium most frequently adopted in environmental sampling for airborne fungi is MEA (malt extract agar) recommended by the ACGIH for its suitability for most fungal growth . DG18 (dichloran glycerol-18), developed in 1980, is suggested for growth at lower water activity (a(w)=0.95) specifically and is not as commonly used in general studies . This investigation collected airborne viable fungi using a single stage/N6 Andersen impactor with MEA and DG18 agar plates attached simultaneously to the same set of samplers . The sampling locations were at 17 sites within a central air-conditioned hospital . After incubation and morphological identification, concentrations of airborne fungi and bacteria were expressed as CFU/m(3) (colony forming units/m(3)) . There are 405 DG18 plates and 378 plates available for statistical analysis . Results show that the airborne fungal concentrations, shown by geometric mean (GM), are higher from the DG18 plates than from the MEA plates . The total fungal concentrations is 68.6 vs 12.94 CFU/m(3), and for Aspergillus spp., the concentration is 1.58 vs 0.72 CFU/m(3); for Penicillium spp., 3.37 vs 0.71; and for yeast, 5.09 vs 0.49 CFU/m(3) . In addition, the number of different genera present is greater on the DG18 plates than on the MEA plates, on average, 2.85 types vs 1.72 . This study suggests that in a hospital environment with 24-h, central air conditioning, DG18 plates appear to be more effective in collecting more fungal colonies in terms of both quantity and types of genera . Such a finding is presumed to be attributed to the characteristic of DG18 in slowing colony growth so that the dominating genus will not over occupy the culture plate surface before the less competitive genus can fully develop . Future studies on related biological mechanisms are essential to conclude whether the above results sustain when sampling is conducted in other environments . Proc Natl Acad Sci U S A, 2000 Feb 15, 97(4), 1665 - 70 In vitro cloning of complex mixtures of DNA on microbeads: physical separation of differentially expressed cDNAs; Brenner S et al.; We describe a method for cloning nucleic acid molecules onto the surfaces of 5-micrometer microbeads rather than in biological hosts . A unique tag sequence is attached to each molecule, and the tagged library is amplified . Unique tagging of the molecules is achieved by sampling a small fraction (1%) of a very large repertoire of tag sequences . The resulting library is hybridized to microbeads that each carry approximately 10(6) strands complementary to one of the tags . About 10(5) copies of each molecule are collected on each microbead . Because such clones are segregated on microbeads, they can be operated on simultaneously and then assayed separately . To demonstrate the utility of this approach, we show how to label and extract microbeads bearing clones differentially expressed between two libraries by using a fluorescence-activated cell sorter (FACS) . Because no prior information about the cloned molecules is required, this process is obviously useful where sequence databases are incomplete or nonexistent . More importantly, the process also permits the isolation of clones that are expressed only in given tissues or that are differentially expressed between normal and diseased states . Such clones then may be spotted on much more cost-effective, tissue- or disease-directed, low-density planar microarrays. Am Surg, 2000 Feb, 66(2), 126 - 32 Sepsis syndrome; Fry DE; A clinical syndrome including fever, leukocytosis, elevated cardiac output, and reduced systemic vascular resistance has been associated with severe infection (i.e., sepsis) . However, during the last 15 years, many patients have demonstrated all of the findings that have traditionally been associated with "sepsis" but have not had demonstrated sources of infection . This led to the term "sepsis syndrome" to refer to that population of patients who appeared to have a physiologic and metabolic response associated with, but who did not have, severe infection . More commonly called the systemic inflammatory response syndrome (SIRS), the sepsis syndrome is now associated with the nonspecific systemic activation of the human inflammatory cascade by any of a number of clinical events . The management of the SIRS patient has been ineffective because of incomplete definition of the mechanisms responsible for the syndrome . It is argued that all of the biological mechanisms that are operative in a simple wound and are beneficial are negative for the host when activated systemically . Thus, SIRS is seen in three separate scenarios at present: (1) invasive infection; (2) dissemination of microbes secondary to failure of host defense mechanisms; and (3) severe activation of inflammation by injury, shock, severe soft tissue inflammation, and other noninfectious but proinflammatory events . Newer treatment strategies will need to focus not on the inciting event itself but on better control of the complex responses of the host. Am Surg, 2000 Feb, 66(2), 117 - 25 Diagnosis and treatment of opportunistic infections in immunocompromised surgical patients; Dunn DL; The advent of successful therapy for patients who suffer many types of organ dysfunction and failure, malignancies, and acquired immunodeficiency syndrome has led to the concurrent threat of infection due to a wide array of pathogens, particularly opportunistic microbes that rarely cause disease under routine circumstances . Among patients who are subjected to extreme degrees of immunosuppression, almost any type of bacterial, fungal, viral, protozoal, or parasitic organism can exhibit pathogenic potential and lead to devastating consequences for the host . Immunosuppressive drug therapy for the purpose of organ allograft maintenance, cancer chemotherapy, or the human immunodeficiency virus exerts potent effects upon cellular immunity . Therefore, although these groups of patients are more susceptible to all types of infectious disease processes, infections due to those pathogens that require a component of cellular immunity for their eradication, such as fungi and viruses, occur at a higher frequency than that observed among normal individuals . Of critical importance, all types of infections are associated with higher rates of morbidity and mortality in immunosuppressed patients . Currently, improved diagnostic techniques and new treatment modalities have rendered many serious infections, for which suitable therapy previously did not exist, amenable to treatment . Because of the large number of immunosuppressed patients who now lead highly productive lives, it is important for the surgical practitioner to become familiar with the modalities currently available to precisely diagnose and effectively treat opportunistic infections in immunocompromised surgical patients. J Lab Clin Med, 2000 Feb, 135(2), 129 - 38 Cytokines as targets of immunotherapy in bacterial pneumonia; Standiford TJ et al.; The generation of a vigorous inflammatory response is essential for the rapid clearance of microbes from the alveolar space . The magnitude of the inflammatory response is tightly controlled by host-derived cytokines, which mediate lung inflammation by serving as leukocyte chemoattractants, leukocyte activating factors, or afferent signals in the induction or regulation of other effector molecules . In this article the role of specific cytokines in lung innate immunity will be reviewed . Future directions regarding the use of specific forms of immunotherapy, including compartmentalized cytokine delivery with gene therapy as adjuvant therapy in the treatment of pneumonia, will be explored. Parasitol Today, 2000 Mar, 16(3), 114 - 8 Tsetse--A haven for microorganisms; Aksoy S; Arthropods are involved in the transmission of parasitic and viral agents that cause devastating diseases in animals and plants . Effective control strategies for many of these diseases still rely on the elimination or reduction of vector insect populations . In addition to these pathogenic organisms, arthropods are rich in microbes that are symbiotic in their associations and are often necessary for the fecundity and viability of their hosts . Because the viability of the host often depends on these obligate symbionts, and because these organisms often live in close proximity to disease-causing pathogens, they have been of interest to applied biologists as a potential means to genetically manipulate populations of pest species . As knowledge on these symbiotic associations accumulates from distantly related insect taxa, conserved mechanisms for their transmission and evolutionary histories are beginning to emerge . Here, Serap Aksoy summarizes current knowledge on the functional and evolutionary biology of the multiple symbionts harbored in the medically and agriculturally important insect group, tsetse, and their potential role in the control of trypanosomiasis. Wei Sheng Yan Jiu, 1998 May, 27(3), 164 - 6 {Effect of particles size calibration on air microbe sampling}; Hu Q; The airborne bacterial particles number sampled with 6 grades Andersen sampler at Xidan in Beijing from 1987 to 1988 was calibrated by the alive bioparticles calibration formula . The results showed that the average number of airborne bacterial particles sampled in 3 minutes and in 84.9 L air was 257, but it is 315 when calibrated . The later was higher than the former (t = 2.012, P < 0.05) . The average number sampled with the 1st grade sampler (> 8.2 microns) was 98, but it is 136 when calibrated, and the difference was obviously (t = 2.409, P < 0.05) . There was no obvious difference between the number sampled with the rest grades sampler (2nd-6th) and when calibrated (t = 1.701-0.026, P > 0.05) . The effects of calibration on the particles size distribution and its concentration day's change were not obviously. Shi Yan Sheng Wu Xue Bao, 1997 Mar, 30(1), 73 - 81 {Studies on cell signaling immunomodulated murine peritoneal suppressor macrophages: LPS and PMA mediate the activation of RAF-1, MAPK p44 and MAPK p42 and p38 MAPK}; Chang ZL et al.; Monocytes-macrophages which serve as host immune cells to kill pathogens can often be "activated" after exposing to viruses, bacteria, cytokines as well as chemical substances, However, it is paradoxical that highly activated macrophages can be induced to become the suppressor ones by live microbes, microbial products, tumor, and autoimmune disease, although the mechanism remains unknown . Our previous experimental studies have shown that immuno-suppressor activities of suppressor macrophages on T, B and NK cells can be prevented by the treatment with LPS or supernatant in vitro from mitogen-stimulated lymphocytes, while, at the same time, the tumoricidal activities of those macrophages can be kept or even enhanced following the same treatment . This phenomenon was then termed as "immune modulation" For the understanding of its mechanism, we are now undertaking signal transduction in modulated macrophages . Since mitogen-activated protein kinase (MAPK) is an integration point of different signal transduction pathways, its cascade and regulation of activation are being investigated extensively by the assay of electrophoresis mobility shift . Recent results suggested that interaction of ligand-receptor triggers protein tyrosine kinase(PTK) activation leading to Ras-GTP binding with Raf-1 to phosphorylate MAPK kinase (MAPKK), the specific activator of MAPK . It is reported that PKC-alpha can directly phosphorylate or activate Raf-1 in NIH3 T3 cells . Raf-1 (74 KDa), with an intrinsic serine (Ser)-threonine (The) kinase activity, becomes hyperphosphorylated after activation which can be followed by gel mobility shift test . It has also been shown that a variety of extracellular factors stimulate a pair of MAPK p44 and MAPK p42 of MAPK family members . A significant property of activation of ERK 1 and ERK 2 is the requirement for the phosphorylation of both Thr-183 and Tyr-185 (at TEY motif) within in its protein kinase subdomain VIII . More recently, two other MAPK subtypes, p38 MAPK (mammalian equivalents of HOG1 in yeast) and JNK MAPK have been discovered . The requirement for activation of p38 MAPK for both Thr-180 and Tyr-182 (at TGY motif) has been shown . p38 MAPK is important in certain transcriptional regulatory pathways, since it can phosphorylate the following transcriptional factors: 1) Elk at Ser 383/389 for binding with SRE motif; 2) . ATF 2 at Ser 69/71, forming a complex with Myc for DNA binding at CRE motif; 3) Max at Ser-62 to combine DNA of E-Box motif . p38 MAPK can be activated by LPS, inflammatory cytokines, such as TNF and IL-1, osmolarity . To examine the possibility that whether activation of Raf-1 and ERK 1, ERK2 and p38 MAPK can be regulated directly or/and differently by PKC and PKA pathways, herbimycin A (Ki = 0.9 mumol/L), a potent PTK inhibitor (J . Immunol . 155:3944-4003, 1995) at 2 mumol/L concentration was utilized to block Ras/Raf-1/MAPK cascade . After pre-incubation of macrophages with herbimycin A for 30 min or 90 min, cells were treated with LPS (10 micrograms/ml) and PMA (100 nmol/L) for 15 min . No inhibition of phosphorylation of Raf-1, MAPK p44 and MAPK p42 in response to LPS and PMA was observed (Fig . 1 and 3) . However, forskolin, a cAMP inducer for protein kinase A (PKA) activation, inhibited the phosphorylation of LPS- and PMA-stimulated Raf-1, MAPK p44 and MAPK p42 (Fig . 2 and 4) . Similarly, in agreement with a very recent report from David, M et al in NIH, in which they indicated that forskolin (30 mumol/L) inhibited IFN-beta-stimulated ERK activity by U 266 cells (J . Biol . Chem . 271: 4585-4588 1996), we found that the levels of phosphorylations of Raf-1 and ERK1 and ERK2 were declined when forskolin (30 mumol/L) was added to macrophages for 20 min at 37 degrees C prior to the stimulation by LPS and PMA . Interestingly, under the same condition, forskolin (30 mumol/L) stimulated the phosphorylation of LPS- and PMA-triggered p38 MAPK of murine peritoneal suppressor macrophages, suggesting that activatio J Comp Neurol, 2000 Feb 14, 417(3), 299 - 314 Sensory neuroanatomy of a passively ingested nematode parasite, Haemonchus contortus: amphidial neurons of the first stage larva; Li J et al.; When infective larvae of Haemonchus contortus (a highly pathogenic, economically important, gastric parasite of ruminants) are ingested by grazing hosts, they are exposed to environmental changes in the rumen, which stimulate resumption of development . Presumably, resumption is controlled by sensory neurons in sensilla known as amphids . Neuronal function can be determined by ablation of specifically recognized neurons in hatchling larvae (L1) in which neuronal cell bodies are easily visualized using differential interference microscopy . Using three-dimensional reconstructions from electron micrographs of serial transverse sections, amphidial structure of the L1 is described . Each amphid of H . contortus is innervated by 12 neurons . The ciliated dendritic processes of 10 neurons lie in the amphidial channel . Three of these end in double processes, resulting in 13 sensory cilia in the channel . One process, that of the so-called finger cell, ends in a number of digitiform projections . Another specialized dendrite enters the amphidial channel, but leaves it to end within the sheath cell, a hollow, flask-shaped cell that forms the base of the amphidial channel . Although not flattened, this process is otherwise similar to the wing cells in Caenorhabditis elegans; we consider it AWC of this group . Two other neurons, ASA and ADB, appear to be homologs of wing cells AWA and AWB in C . elegans, although they end as ciliated processes in the amphidial channel, rather than as flattened endings seen in C . elegans . Each of the 12 amphidial neurons was traced to its cell body in the lateral ganglion, posterior to the worm's nerve ring . The positions of these bodies were similar to their counterparts in C . elegans; they were named accordingly . A map for identifying the amphidial cell bodies in the living L1 was prepared, so that laser microbeam ablation studies can be conducted . These will determine which neurons are involved in the infective process, as well as others important in establishing the host-parasite relationship. Proc Natl Acad Sci U S A, 2000 Feb 29, 97(5), 2099 - 104 Induction of a bystander mutagenic effect of alpha particles in mammalian cells; Zhou H et al.; Ever since the discovery of X-rays was made by Rontgen more than a hundred years ago, it has always been accepted that the deleterious effects of ionizing radiation such as mutation and carcinogenesis are attributable mainly to direct damage to DNA . Although evidence based on microdosimetric estimation in support of a bystander effect appears to be consistent, direct proof of such extranuclear/extracellular effects are limited . Using a precision charged particle microbeam, we show here that irradiation of 20% of randomly selected A(L) cells with 20 alpha particles each results in a mutant fraction that is 3-fold higher than expected, assuming no bystander modulation effect . Furthermore, analysis by multiplex PCR shows that the types of mutants induced are significantly different from those of spontaneous origin . Pretreatment of cells with the radical scavenger DMSO had no effect on the mutagenic incidence . In contrast, cells pretreated with a 40 microM dose of lindane, which inhibits cell-cell communication, significantly decreased the mutant yield . The doses of DMSO and lindane used in these experiments are nontoxic and nonmutagenic . We further examined the mutagenic yield when 5-10% of randomly selected cells were irradiated with 20 alpha particles each . Results showed, likewise, a higher mutant yield than expected assuming no bystander effects . Our studies provide clear evidence that irradiated cells can induce a bystander mutagenic response in neighboring cells not directly traversed by alpha particles and that cell-cell communication process play a critical role in mediating the bystander phenomenon. J Biomater Sci Polym Ed, 2000, 11(1), 13 - 25 Protein A immobilization and HIgG adsorption onto porous/nonporous and swellable HEMA-incorporated polyEGDMA microspheres; Ayhan H et al.; Both non swellable and swellable poly(EGDMA/HEMA) microbeads were produced by suspension copolymerization . These microbeads were modified by immobilization of a spacer-arm (hexamethylene diamine (HMDA)) and protein A . The optimal values for modifications were as follows: sodium periodate concentration, 1.0 mgml(-1); HMDA concentration, 4 mgml(-1); and glutaraldehyde concentration, 0.070 microgml(-1) . Adsorption of protein A onto the plain and periodate oxidized poly(EGDMA/HEMA) microbeads were very close to each other, and were 0.01-0.02 mg protein A on the 1-g Microbeads I and II, respectively . Protein A immobilization on poly(EGDMA/HEMA) microbeads were studied at different temperatures, times, and pHs using single protein solution containing different amounts of proteins . The optimal values for immobilization were as follows: the initial protein A concentration, 0.1 mgml(-1); temperature, 25 degrees C; pH, 9.5; and immobilization time, 120 min . Incorporation of protein A resulted in 1.420 and 1.825 mg protein A on the 1-g Microbeads I and II, respectively . HIgG adsorption capacity on the protein A-incorporated poly(EGDMA/HEMA) microbeads is 27 and 35 mg HIgGg(-1) polymer for Microbeads I and II, respectively. Biosci Biotechnol Biochem, 1999 Nov, 63(11), 1953 - 8 5-Hydroxy-4-oxo-L-norvaline depletes intracellular glutathione: a new modulator of drug resistance; Tagashira M et al.; To search for compounds that reverse the drug resistance induced by glutathione (GSH), an original screening system to detect intracellular GSH depleters was established . Among 8843 microbes derived from the soil samples tested, the extracts of two Streptomyces species named KS6701 and KS8846, lowered the intracellular GSH level of Saccharomyces cerevisiae 5 x 47 . From both the microbes, 5-hydroxy-4-oxo-L-norvaline (HON) was isolated as the active compound . At a concentration of 50-100 micrograms/ml, HON also decreased the GSH/protein level of the human ovarian tumor cell line, 2008/C13*5.25 and reversed its resistance to cisplatin . We also investigated the mechanism of the depletion . HON had little effect on gamma-glutamylcysteine synthetase (gamma-GCS) or glutathione synthetase, but HON decreased the quantity of thiol substances when it was spontaneously reacted with them . This suggested that the GSH depletion by HON occurred through a mechanism different from that of buthionine sulfoximine, a selective gamma-GCS inhibitor. Artif Cells Blood Substit Immobil Biotechnol, 2000 Jan, 28(1), 65 - 77 Evaluation of heparin immobilized chitosan-PEG microbeads for charcoal encapsulation and endotoxin removal; Chandy T et al.; A technique is described to encapsulate activated charcoal for hemoperfusion to be used in an artificial liver support . Activated charcoal was encapsulated within chitosan-PEG matrix and subsequently surface modified with PGE1 or heparin (hep-AC-PEGCB) via the glutaraldehyde functionalities . This novel matrix was used as the supports for perfusion of endotoxin, under a flow rate of 30 ml/mt . Endotoxin adsorption was quantitatively measured by the method of Limulus Amebocyte lysate test . It seems, the hep-AC-PEGCB may be a good adsorbent system for the removal of toxic endotoxin, and the system may be useful for detoxification of blood . The hep-AC-PEGCB matrix had improved biocompatibility as demonstrated from their hemolytic potential and charcoal release . However, further studies are needed to determine their behaviour under clinical conditions. Biol Trace Elem Res, 1999 Winter, 71-72, 77 - 82 An external ion microbeam for studies of biological samples; Sakai T et al.; A new external ion-beam system was developed and combined with a light ion-microbeam system in JAERI Takasaki . The system is designed for micro-particle-induced X-ray emission analysis of biological samples in air environment with 1 micron spatial resolution . One of the most serious problems in keeping such a high spatial resolution is multiple scattering in a beam exit window . Thin Kapton film (7.5 microns thick) was adopted as the exit window as well as a sample backing foil to minimize the distance between the film and samples . The lifetime of the foil under ion irradiation and spatial resolution of the external microbeam were investigated . The results shows that the film can endure sufficient long-time irradiation to take elemental maps and the resolution can be kept nearly 1 micron. Biol Trace Elem Res, 1999 Winter, 71-72, 57 - 64 Microbeam PIXE analysis of Na, K, Mg, and Ca in severely damaged cardiac tissue; Verhoef BA et al.; The aim of the study was to measure with microbeam PIXE elements such as Na, K, Mg, and Ca in cardiac tissue after various treatments in vivo, which affect the cardiomyocyte integrity . It was assumed that local deviations from normal electrolyte levels indicate the degree of cardiac damage . The first step in this feasibility study was comparison of severely damaged cardiac tissues with controls . Severe cardiac damage was introduced by the so-called Ca paradox . Experiments were performed with isolated rat hearts, perfused retrogradely with an oxygenated crystalloid buffer . Results indicated that severe cardiac damage was accompanied with almost complete disappearance of the normal intracellular electrolyte composition as a result of the loss of membrane integrity . Identifications of smaller and more locally present ischemic damages on basis of altered electrolyte levels appeared to be feasible . However, the prerequisite was that the mobility of electrolytes be kept under control during tissue sampling and sample preparation, when physiological mechanisms stop to maintain gradients. J Cardiovasc Pharmacol, 2000 Feb, 35(2), 294 - 301 Sarpogrelate, a selective 5-HT2A serotonergic receptor antagonist, inhibits serotonin-induced coronary artery spasm in a porcine model; Miyata K et al.; Serotonin is one of the most important vasoactive substances and has been implicated in the pathogenesis of coronary artery spasm and of acute coronary syndrome . We have recently demonstrated that local and long-term treatment with interleukin-1beta(IL-1beta) causes coronary arteriosclerotic changes and hyperconstrictive responses to serotonin in pigs in vivo . However, it remains to be examined which serotonergic (5-HT) receptor subtype mediates coronary spasm and whether alterations in serotonergic receptors are involved in the abnormality . In this study, we examined the inhibitory effect of sarpogrelate, a selective 5-HT2A serotonergic receptor antagonist, on the serotonin-induced coronary spasm as well as the possible alterations of serotonergic receptors in our porcine model . A segment of the porcine coronary artery was carefully dissected and aseptically wrapped with cotton mesh absorbing IL-1beta-bound microbeads from the adventitia . Two weeks after the procedure, angiographic study was performed, followed by binding assay for 5-HT1B and 5-HT2A serotonergic receptors and reverse transcription-polymerase chain reaction (RT-PCR) analysis for mRNA of those receptors . Angiographic study showed coronary vasospastic responses to serotonin at the IL-1beta-treated site . Sarpogrelate dose-dependently inhibited the serotonin-induced coronary spasm, but it did not affect the prostaglandin F2alpha-induced vasoconstriction . Radiolabeled receptor-binding assay showed that receptor affinity or receptor number of the 5-HT1B, or 5-HT2A receptors did not differ significantly between the spastic and the control sites . Furthermore, RT-PCR analysis showed that the expression of neither 5-HT2A nor 5-HT1B receptor mRNA was significantly altered at the spastic site . These results indicate that serotonin-induced coronary spasm is mediated primarily by 5-HT2A receptor in our porcine model, although the 5-HT2A receptor was not up-regulated, suggesting that alteration in the signal-transduction pathway for vascular smooth muscle contraction beyond the 5-HT2A receptor plays a primary role in the pathogenesis of coronary spasm in our porcine model. Fungal Genet Biol, 1999 Dec, 28(3), 214 - 26 Major surface glycoprotein genes from Pneumocystis carinii f . sp . ratti; Schaffzin JK et al.; Pneumocystis carinii occurs in a variety of mammals, each of which harbors one or more genetically distinct "special forms" of the microbe . Laboratory rats can be infected by two special forms, P . carinii f . sp . ratti and P . carinii f . sp . carinii . P . carinii f . sp . carinii has a variable antigen, the major surface glycoprotein (MSG), the expression of which is controlled by genetic recombination . Recombination may involve the CRJE, a 23-bp DNA sequence element invariant among P . carinii f . sp . carinii MSG genes . To better understand the role of the CRJE in MSG gene expression and to explore the possible role of MSG in P . carinii infection in rats, P . carinii f . sp . ratti MSG genes were studied . These genes were found to be related to MSG genes of P . carinii f . sp . carinii, but less so than MSG genes from P . carinii f . sp . carinii are to each other . P . carinii f . sp . ratti MSG genes were present throughout the genome and were expressed as an abundant mRNA species slightly smaller than that found in P . carinii f . sp . carinii . P . carinii f . sp . ratti MSG transcripts included a CRJE-like sequence only 78% identical to the CRJE of P . carinii f . sp . carinii . Comparison of MSG proteins from the two rat special forms of P . carinii to those from human, ferret, and mouse P . carinii did not support the hypothesis that growth in the rat lung requires certain primary MSG peptide sequences . BMJ, 2000 Feb 12, 320(7232), 412 - 7 Exposure to foodborne and orofecal microbes versus airborne viruses in relation to atopy and allergic asthma: epidemiological study; Matricardi PM et al.; OBJECTIVE: To investigate if markers of exposure to foodborne and orofecal microbes versus airborne viruses are associated with atopy and respiratory allergies . DESIGN: Retrospective case-control study . PARTICIPANTS: 240 atopic cases and 240 non-atopic controls from a population sample of 1659 participants, all Italian male cadets aged 17-24 . SETTING: Air force school in Caserta, Italy . MAIN OUTCOME MEASURES: Serology for Toxoplasma gondii, Helicobacter pylori, hepatitis A virus, measles, mumps, rubella, chickenpox, cytomegalovirus, and herpes simplex virus type 1; skin sensitisation and IgE antibodies to relevant airborne allergens; total IgE concentration; and diagnosis of allergic asthma or rhinitis . RESULTS: Compared with controls there was a lower prevalence of T gondii (26% v 18%, P=0.027), hepatitis A virus (30% v 16%, P=0.004), and H pylori (18% v 15%, P=0.325) in atopic participants . Adjusted odds ratios of atopy decreased with a gradient of exposure to H pylori, T gondii, and hepatitis A virus (none, odds ratio 1; one, 0 . 70; two or three, 0.37; P for trend=0.000045) but not with cumulative exposure to the other viruses . Conversely, total IgE concentration was not independently associated with any infection . Allergic asthma was rare (1/245, 0.4%) and allergic rhinitis infrequent (16/245, 7%) among the participants (245/1659) exposed to at least two orofecal and foodborne infections (H pylori, T gondii, hepatitis A virus) . CONCLUSION: Respiratory allergy is less frequent in people heavily exposed to orofecal and foodborne microbes . Hygiene and a westernised, semisterile diet may facilitate atopy by influencing the overall pattern of commensals and pathogens that stimulate the gut associated lymphoid tissue thus contributing to the epidemic of allergic asthma and rhinitis in developed countries. Naturwissenschaften, 2000 Jan, 87(1), 1 - 11 Symbiosis and pathogenesis: evolution of the microbe-host interaction; Steinert M et al.; Symbiotic and pathogenic bacteria have in common that they live in or on host organisms or host cells . To make a successful living in eukaryotic hosts, bacteria must possess the traits to recognize a given host and establish adherence . When the bacterial location is internal or intracellular, they must further have the ability to invade, to establish a niche, and finally to multiply within a host . The underlying mechanisms which allow this form of existence show similarities between symbiotic and pathogenic bacteria . The final outcome, however, may result in a wide spectrum of consequences for the host ranging from the acquisition of novel metabolic pathways to damage or death . Despite the vastly different forms of interactions, symbiotic and pathogenic bacteria have in common that they are adapted to a particular environmental niche represented by the host organism or compartment thereof . This contribution reviews the evolutionary forces which have shaped the microbial-host interactions . Particular emphasis is placed on the genetic and molecular mechanisms that drive bacterial evolution in response to the selective pressures of the host environment. Phys Med Biol, 2000 Jan, 45(1), 103 - 19 Oxygen consumption through metabolism and photodynamic reactions in cells cultured on microbeads; Schunck T et al.; Oxygen consumption by cultured cells, through metabolism and photosensitization reactions, has been calculated theoretically . From this result, we have derived the partial oxygen pressure PO2 in the perfusion medium flowing across sensitized cultured cells during photodynamic experiments . The PO2 variations in the perfusate during light irradiation are related to the rate of oxygen consumption through photoreactions, and to the number of cells killed per mole of oxygen consumed through metabolic processes . After irradiation, the reduced metabolic oxygen consumption yields information on the cell death rate, and on the photodynamic cell killing efficiency . The aim of this paper is to present an experimental set-up and the corresponding theoretical model that allows us to control the photodynamic efficiency for a given cell-sensitizer pair, under well defined and controlled conditions of irradiation and oxygen supply . To demonstrate the usefulness of the methodology described, CHO cells cultured on microbeads were sensitized with pheophorbide a and irradiated with different light fluence rates . The results obtained, i.e . oxygen consumption of about 0.1 microM s(-1) m(-3) under a light fluence rate of 1 W m(-2), 10(5) cells killed per mole of oxygen consumed and a decay rate of about 1 h(-1) of living cells after irradiation, are in good agreement with the theoretical predictions and with previously published data. Med Phys, 2000 Jan, 27(1), 239 - 44 Improved spatial resolution by MOSFET dosimetry of an x-ray microbeam; Kaplan GI et al.; Measurement of the lateral profile of the dose distribution across a narrow x-ray microbeam requires a dosimeter with a micron resolution . We investigated the use of a MOSFET dosimeter in an "edge-on" orientation with the gate insulating oxide layer parallel to the direction of the beam . We compared results using this technique to Gafchromic film measurements of a 200 micrometer wide planar x-ray microbeam . The microbeam was obtained by using a vernier micrometer-driven miniature collimator attached to a Therapax DXT300 x-ray machine operated at 100 kVp . The "edge-on" application allows utilization of the ultra thin sensitive volume of the MOSFET detector . Spatial resolution of both the MOSFET and Gafchromic film dosimeters appeared to be of about 1 micrometer . The MOSFET dosimeter appeared to provide more uniform dose profiles with the advantage of on-line measurements. Mol Plant Microbe Interact, 2000 Feb, 13(2), 151 - 8 Hypaphorine from the ectomycorrhizal fungus Pisolithus tinctorius counteracts activities of indole-3-acetic acid and ethylene but not synthetic auxins in eucalypt seedlings; Ditengou FA et al.; Very little is known about the molecules regulating the interaction between plants and ectomycorrhizal fungi during root colonization . The role of fungal auxin in ectomycorrhiza has repeatedly been suggested and questioned, suggesting that, if fungal auxin controls some steps of colonized root development, its activity might be tightly controlled in time and in space by plant and/or fungal regulatory mechanisms . We demonstrate that fungal hypaphorine, the betaine of tryptophan, counteracts the activity of indole-3-acetic acid (IAA) on eucalypt tap root elongation but does not affect the activity of the IAA analogs 2,4-D ((2,4-dichlorophenoxy)acetic acid) or NAA (1-naphthaleneacetic acid) . These data suggest that IAA and hypaphorine interact during the very early steps of the IAA perception or signal transduction pathway . Furthermore, while seedling treatment with 1-amincocyclopropane-1-carboxylic acid (ACC), the precursor of ethylene, results in formation of a hypocotyl apical hook, hypaphorine application as well as root colonization by Pisolithus tinctorius, a hypaphorine-accumulating ectomycorrhizal fungus, stimulated hook opening . Hypaphorine counteraction with ACC is likely a consequence of hypaphorine interaction with IAA . In most plant-microbe interactions studied, the interactions result in increased auxin synthesis or auxin accumulation in plant tissues . The P . tinctorius / eucalypt interaction is intriguing because in this interaction the microbe down-regulates the auxin activity in the host plant . Hypaphorine might be the first specific IAA antagonist identified. Bioorg Med Chem, 1999 Dec, 7(12), 3025 - 38 Syntheses and studies of multiwarhead siderophore-5-fluorouridine conjugates; Lu Y et al.; Siderophores are microbial iron chelating agents that sequester physiologically essential iron for microbes . Conjugation of drugs to siderophores allows use of active iron transport for microbially directed drug delivery . Syntheses and biological studies are described of the first multidrug isocyanurate-based siderophore analogues separately containing one, two, and three 5-fluorouridine (5-FU) derivatives as the drug component . The results indicate that a single siderophore can be used to deliver multiple drugs to target pathogenic microorganisms. Cytometry, 2000 Jan 1, 39(1), 56 - 66 A new method for the simultaneous analysis of growth and death of immunophenotypically defined cells in culture; Prieto A et al.; BACKGROUND: Internal standards have been used in flow cytometry methods to enumerate lymphoid subsets and hemopoietic progenitor cells ex vivo . However, the currently available methods cannot be readily applied to the analysis of cultured cells because of the frequent occurrence of cell death during in vitro assays . METHODS: This paper reports a new method for the enumeration of both viable and nonviable cells in culture . Cells were counted with the aid of an internal reference standard of microbeads, and live versus dead cell discrimination was performed using 7-amino-actinomycin D which allows the double staining of surface antigens . RESULTS: The method is more precise, accurate and sensitive than either conventional light microscopy-based or automated cell counting . Additionally, it may be used to accurately measure the number of apoptotic cells in a culture . RESULTS: Through the enumeration of surviving cells it is demonstrated that, when applied to the study of mitogen-activated T lymphocytes, current flow cytometry techniques (which do not use internal standards) for the study of the viability and apoptosis overestimate the fraction of viable cells and underestimate both the fraction of dead and apoptotic cells . CONCLUSIONS: The new method overcomes these limitations and is of use in the in vitro study of cell growth and apoptosis . Proc Natl Acad Sci U S A, 2000 Feb 1, 97(3), 1247 - 51 A habitat for psychrophiles in deep Antarctic ice; Price PB; Microbes, some of which may be viable, have been found in ice cores drilled at Vostok Station at depths down to approximately 3,600 m, close to the surface of the huge subglacial Lake Vostok . Two types of ice have been found . The upper 3,500 m comprises glacial ice containing traces of nutrients of aeolian origin including sulfuric acid, nitric acid, methanosulfonic acid (MSA), formic acid, sea salts, and mineral grains . Ice below approximately 3,500 m comprises refrozen water from Lake Vostok, accreted to the bottom of the glacial ice . Nutrients in the accretion ice include salts and dissolved organic carbon . There is great interest in searching for living microbes and especially for new species in deepest Antarctic ice . I propose a habitat consisting of interconnected liquid veins along three-grain boundaries in ice in which psychrophilic bacteria can move and obtain energy and carbon from ions in solution . In the accretion ice, with an age of a few 10(4) years and a temperature a few degrees below freezing, the carbon and energy sources in the veins can maintain significant numbers of cells per cubic centimeter that are metabolizing but not multiplying . In the 4 x 10(5)-year-old colder glacial ice, at least 1 cell per cm(3) in acid veins can be maintained . With fluorescence microscopy tuned to detect NADH in live organisms, motile bacteria could be detected by direct scanning of the veins in ice samples. J Clin Microbiol, 2000 Feb, 38(2), 807 - 13 Pathogenic clones versus environmentally driven population increase: analysis of an epidemic of the human fungal pathogen Coccidioides immitis; Fisher MC et al.; For many pathogenic microbes that utilize mainly asexual modes of reproduction, it is unknown whether epidemics are due to either the emergence of pathogenic clones or environmentally determined increases in the population size of the organism . Descriptions of the genetic structures of epidemic populations, in conjunction with analyses of key environmental variables, are able to distinguish between these competing hypotheses . A major epidemic of coccidioidomycosis (etiologic agent, Coccidioides immitis) occurred between 1991 and 1994 in central California, representing an 11-fold increase above the mean number of cases reported from 1955 to 1990 . Molecular analyses showed extensive genetic diversity, a lack of linkage disequilibria, and little phylogenetic structure, demonstrating that a newly pathogenic strain was not responsible for the observed epidemic . Epidemiological analyses showed that morbidity caused by C . immitis was best explained by the interaction between two variables, the lengths of droughts preceding epidemics and the amounts of rainfall . This shows that the principal factors governing this epidemic of C . immitis are environmental and not genetic . An important implication of this result is that the periodicity of cyclical environmental factors regulates the population size of C . immitis and is instrumental in determining the size of epidemics . This knowledge provides an important tool for predicting outbreaks of this pathogen, as well as a general framework that may be applied to determine the causes of epidemics of other fungal diseases. J Microbiol Immunol Infect, 1999 Dec, 32(4), 269 - 77 {The effect of thermal power plant on microbial ecology and environmental quality}; Yang SS et al.; To investigate the effect of thermal power plant on the microbial ecology and the environmental quality, the Hsieh-Ho Thermal Power Plant was chosen and the populations of microbes including bacteria, actinomycetes, fungi, and cellulolytic, phosphate-solubilizing and nitrogen-fixing microbes were selected as the parameters of microbial ecology . The pH values of the soil sample collected from inside and outside of the plant were 5.2-6.2 and 4.0-5.3, respectively . Moisture content in plant area was lower than that in the surrounding area . Microbial populations of the topsoils were higher than those of the subsoils . Each gram of soil contained 3.64 x 10(4)-5.16 x 10(7) colonies of bacteria, 1.75 x 10(3)-1.10 x 10(6) colonies of actinomycetes and 6.72 x 10(3)-8.78 x 10(6) colonies of fungi in the plant area; while they were 5.52 x 10(4)-2.14 x 10(7), 8.26 x 10(3)-7.25 x 10(5) and 3.49 x 10(3)-2.74 x 10(6) colonies of bacteria, actinomycetes and fungi, respectively, in the surrounding area . The effect of seasonal change on microbial populations was not significant . The ratio of cellulolytic, phosphate-solubilizing and nitrogen-fixing microbes to the total count in the plant area was also higher than that in the surrounding area, and some of them had significant differences . From the statistical analysis, the effect of thermal power generator on the population and distribution of microbes was significantly different. Adv Perit Dial, 1998, 14, 158 - 63 Sphingosine and sphinganine levels in human mesothelial cells in vitro as a potential index of signal transduction pathways impacted by microbes and osmolality; Nowak DM et al.; Sphingolipids are emerging as important regulators of mammalian cell biology . In this study, the contents of six separate preparations of human omental mesothelial cells in vitro were examined for free sphingosine and sphinganine, and for the total levels of these sphingoid bases in ceramide-containing sphingolipids . Two high-performance liquid chromatography (HPLC) methods for determination of sphingoid base levels in cultured cells were compared . The rapid-HPLC method was found to yield the highest recovery of internal standard . Mesothelial cells initially isolated by collagenase digestion of the omentum were found to have higher free- and total-sphingoid base levels than cells isolated by trypsin-EDTA digestion . Use of sphingoid base levels to gain insights into the status of cellular nutrition, inflammation, programmed cell death, exposure to microbial toxins, cytokines, and growth factors within the peritoneum will require a systematic description of sphingolipids in normal, diseased, and dialyzed mesothelium. C R Acad Sci III, 1999 Nov, 322(11), 953 - 8 Development of novel tuberculosis vaccines; Hess J et al.; Efficacious control of tuberculosis (TB), one of the world's major health threats, is best achieved by a combination of chemotherapy and vaccination . The current vaccine, BCG, fails to prevent pulmonary TB in adults, which is the most prevalent form of this disease . Consequently, the design of novel vaccines against TB is urgently required . Because the acquired immune response is mediated by different T-cell sets, an optimal combination of these populations must be stimulated . As one third of the world's population is already infected with Mycobacterium tuberculosis, two types of vaccine may be required: one for eradication of already established infection and the other for prompt combat of invading microbes . A rational judgement on the efficacy of the different types of vaccine currently under development needs to await further evaluation. C R Acad Sci III, 1999 Nov, 322(11), 943 - 51 Vaccination against the major infectious diseases; Plotkin SA; The reputation of vaccination rests on a 200-year-old history of success against major infectious diseases . That success has led to the doctrine of 'for each disease, a vaccine' . Although some diseases have proved frustrating, this doctrine carries considerable truth . However, when one reviews the vaccines now available it is apparent that most successes have been obtained when the microbe has a bacteremic or viremic phase during which it is susceptible to the action of neutralizing antibodies, and before replication in the particular organ to which it is tropic . Poliomyelitis and infections by capsulated bacteria are examples where vaccination has worked efficiently . However, some success has also been achieved against agents replicating on respiratory or gastrointestinal mucosae . Influenza, pertussis and rotavirus vaccines are examples of such agents, against which it has been possible to induce immune responses acting locally as well as systemically . In addition, when bacteria produce disease through exotoxins, purification and chemical or genetic inactivation of those toxins has yielded highly efficacious vaccines . Control of intracellular pathogens has not been achieved, except partly with the BCG vaccine against tuberculosis, and modern efforts are directed towards pathogens against which cellular immune responses are critical . In general, two achievements have been crucial to the success of vaccines: the induction of long-lasting immunological memory in individuals and the stimulation of a herd immunity that enhances control of infectious diseases in populations. Genome Res, 2000 Jan, 10(1), 62 - 71 Simple sequence repeats in Escherichia coli: abundance, distribution, composition, and polymorphism; Gur-Arie R et al.; Computer-based genome-wide screening of the DNA sequence of Escherichia coli strain K12 revealed tens of thousands of tandem simple sequence repeat (SSR) tracts, with motifs ranging from 1 to 6 nucleotides . SSRs were well distributed throughout the genome . Mononucleotide SSRs were over-represented in noncoding regions and under-represented in open reading frames (ORFs) . Nucleotide composition of mono- and dinucleotide SSRs, both in ORFs and in noncoding regions, differed from that of the genomic region in which they occurred, with 93% of all mononucleotide SSRs proving to be of A or T . Computer-based analysis of the fine position of every SSR locus in the noncoding portion of the genome relative to downstream ORFs showed SSRs located in areas that could affect gene regulation . DNA sequences at 14 arbitrarily chosen SSR tracts were compared among E . coli strains . Polymorphisms of SSR copy number were observed at four of seven mononucleotide SSR tracts screened, with all polymorphisms occurring in noncoding regions . SSR polymorphism could prove important as a genome-wide source of variation, both for practical applications (including rapid detection, strain identification, and detection of loci affecting key phenotypes) and for evolutionary adaptation of microbes. Radiology, 2000 Jan, 214(1), 113 - 9 Embolization with radiopaque microbeads of polyacrylonitrile hydrogel: evaluation in swine; Gobin YP et al.; PURPOSE: To perform in vitro and in vivo studies of radiopaque microbeads of polyacrylonitrile (PAN) hydrogel to evaluate their characteristics as embolic material . MATERIALS AND METHODS: PAN microbeads were analyzed in vitro for swelling in different concentrations of contrast material and saline solution and injected through various microcatheters . In three nonsurvival swine, various organs were embolized with PAN microbeads . In eight survival swine, the rete mirabile was embolized with PAN microbeads or polyvinyl alcohol particles . Follow-up angiograms were obtained regularly, and histopathologic analysis was performed at 1 and 6 months . RESULTS: The microbeads were black with a regular shape and smooth surface . They were easily visible in the syringe and easy to inject through the microcatheters . When wet, their diameters increased by 40% . The microbeads were sufficiently radiopaque to be visible in all vascular territories . Vascular occlusion was not permanent, and even when embolization was adequate, some revascularization was detected at 3 months . The microbeads were intact and still radiopaque at 6 months . Histopathologic examination demonstrated variable inflammatory reactions and foreign-body giant cell reaction and no angionecrosis or hemorrhage . CONCLUSION: Because PAN microbeads are biocompatible, radiopaque, and easy to handle during embolization procedures, they have potential as a therapeutic embolic agent. FEMS Microbiol Ecol, 2000 Feb 1, 31(2), 99 - 106 Comparative analyses in aquatic microbial ecology: how far do they go? Gasol JM, Duarte CM. Methodological developments in recent years have led to an increase in empirical databases on the abundance and functions of aquatic microbes, now allowing synthesis studies . Most of these studies have adopted a comparative approach, such that comparative analyses are now available for most aspects of aquatic microbial food webs (more than 50 papers published in the last 15 years) . Some of these analyses apparently yield conflicting results, introducing confusion and unnecessary disputes in the field . We briefly review the comparative analyses so far produced and we highlight generalities, show that some of the perceived discrepancies largely derive from partial analyses of a general underlying trend and formulate predictions based on these general trends that provide new avenues for research. Curr Med Chem, 2000 Feb, 7(2), 159 - 97 Studies and syntheses of siderophores, microbial iron chelators, and analogs as potential drug delivery agents; Roosenberg JM 2nd et al.; Siderophores (microbial iron chelators) play an extremely important role in microbial pathogenicity . Microbial uptake of siderophore-iron complexes through active transport systems allow microbes to survive and proliferate even under iron deficient environments during invasion of a host . Due to their structural complexity, unique iron (III) chelation, acquisition properties, and their therapeutic potential, siderophores have attracted much attention in a broad range of disciplines . Tremendous progress has been made in siderophore syntheses, in determination of the structures and functions of outer membrane receptors (e.g . FhuA and FepA), and in the mechanistic insight into siderophore-iron-mediated active transport processes . One of the important practical applications of this active transport system is development of species-selective active drug transport (the Trojan Horse approach) to potentially treat infections due to drug resistant strains of microbes . Siderophore-drug conjugates have shown great potential in active drug delivery to target pathogenic microbes. J Dermatol, 1999 Nov, 26(11), 757 - 63 New technologies toward dendritic cell-based cancer immunotherapies; Matsue H et al.; Immunologically naive T cells are activated most efficiently or even exclusively by special subsets of antigen presenting cells, termed dendritic cells (DC) . Members of the DC family have been identified in virtually all epithelial tissues that are constantly exposed to environmental antigens or infectious microbes . For example, skin is equipped with at least two members of this family, epidermal Langerhans cells (LC) and dermal DC . DC have been shown to play pathogenic roles in several different inflammatory/immunological disorders and protective roles against infectious pathogenes and cancer development . In this review article, we will overview the recent progress in the development of DC-based immunotherapies for the prevention and treatment of cancers. Nucleic Acids Res, 2001 Jan 1, 29(1), 246 - 54 ARED: human AU-rich element-containing mRNA database reveals an unexpectedly diverse functional repertoire of encoded proteins; Bakheet T et al.; The adenylate uridylate-rich elements (AREs) mediate the rapid turnover of mRNAs encoding proteins that regulate cellular growth and body response to exogenous agents such as microbes, inflammatory and environmental stimuli . However, the full repertoire of ARE-containing mRNAs is unknown . Here, we explore the distribution of AREs in human mRNA sequences . Computational derivation of a 13-bp ARE pattern was performed using multiple expectation maximization for motif elicitations (MEME) and consensus analyses . This pattern was statistically validated for the specificity towards the 3'-untranslated region and not coding region . The computationally derived ARE pattern is the basis of a database which contains non-redundant full-length ARE-mRNAs . The ARE-mRNA database (ARED; reveals that ARE-mRNAs encode a wide repertoire of functionally diverse proteins that belong to different biological processes and are important in several disease states . Cluster analysis was performed using the ARE sequences to demonstrate potential relationships between the type and number of ARE motifs, and the functional characteristics of the proteins. Bone Marrow Transplant, 1999 Dec, 24(12), 1329 - 36 Selective loss of progenitor subsets following clinical CD34+ cell enrichment by magnetic field, magnetic beads or chromatography separation; Johnsen HE et al.; In this preclinical evaluation we have compared the efficacy of three clinical CD34+enrichment procedures with respect to purity, yield and recovery, as well as risk of selective loss of CD34+ lineage-specific subsets . The three devices work by different principles and have several different manipulation steps: The magnetic field separator uses paramagnetic iron-dextran particles; the magnetic microbead selection is based on the advantage of a large surface area for immobilisation of the monoclonal antibody within a very small volume; the original immunoabsorption technique is based on the use of biotinylated antibody applied to a column of avidin-coated sephadex beads . The results of this evaluation gave a median purity 96% (88-98%), 86% (62-97%), and 49% (18-85%), and median yield of 65% (54-100%), 40% (21-74%), and 30% (8-55%), respectively . Subset analysis recognised a selective loss of CD34+/61+ after enrichment, most likely due to class I-II antibodies used for the enrichment step or, alternatively, nonspecific binding of megakaryocytic progenitors . Tumour cell spiking experiments on a clinical scale documented an expected 2-4 log reduction resulting in a number of potentially malignant cells in the CD34 enriched product . Our data support four major conclusions: First, that magnetic field separation is superior to magnetic beads and chromatography selection, mainly due to the risk of cell loss and insufficient recovery with the two latter methods . Second, that late differentiated progenitors with CD34 class III epitopes present are lost during the enrichment procedures . The third major conclusion is that chromatography selection results in a selective loss of CD34bright cells, which are most likely uncommitted early progenitors . This was an unexpected finding which may be a consequence of an imbalance between the strong forces between biotin-avidin and insufficient physical manipulation for CD34+ cell release . Finally, the data document that CD34 selection alone is an inappropriate way to eliminate tumour cells due to the uncontrolled variables and the inconsistent outcome . The only products which can be expected to be purged free of tumour cells are the ones with very minimal (<10-5) contamination in the starting products, ie products documented tumour free with the most sensitive techniques for quantitation . If this is not the case, the optimal purging strategy may be a two-step procedure including CD34 selection and subsequent depletion of the tumour cells in question. Hum Immunol, 1999 Dec, 60(12), 1293 - 302 Flow cytometric detection of HLA antibodies using a spectrum of microbeads; Pei R et al.; We describe here the use of HLA antigen coated beads for specificity and class determination of HLA antibodies by flow cytometry . The HLA specificity of antibodies was determined by use of beads containing eight levels of fluorescence . HLA antigens isolated from eight cultured cells were coated onto these beads so that each bead was the equivalent of one cell . By using four sets of eight beads, an equivalent of 32 cells could be examined in four test tubes . A total of 76 class I and 25 class II specificities could be determined by the 32 class I bead-panel and 32 class II bead-panel used, respectively . We noted no cross-reactivity of reactions between class I and II . The sensitivity of the test was shown to be higher than that of the standard cytotoxicity by dilution experiments and detection of additional cross-reacting antigens . By use of these coated beads, we achieved improved standardized detection of HLA antibodies . Antigen-coated beads have several advantages over the use of spleens or lymphocytes . (a) A highly selected panel of antigens can be routinely used . (b) Class I and class II antibodies can be readily distinguished from each other, even when they are present as mixtures in one serum . (c) Non-HLA antibodies are not detected because the beads do not have any other antigens than HLA on them . (d) The quantity of antigens coated on beads is more uniform than that found in cells from different individuals . (e) Beads are more convenient for storage and daily use. Anal Chem, 1999 Dec 15, 71(24), 5465 - 9 Separating microbes in the manner of molecules . 1 . Capillary electrokinetic approaches; Armstrong DW et al.; Selective, high-efficiency separations of intact bacteria may, in some cases, allow them to be identified and quantified in much the same way that molecules are done today . Two different capillary electrokinetic approaches were utilized . The first approach used a dissolved polymer-based CE separation that may be affected by size and shape considerations . Another approach uses capillary isoelectric focusing to separate bacteria by their surface charge or isoelectric point . Good peak shapes and extremely high efficiencies are observed (up to approximately 1,600,000 theoretical plates/m) . Careful sample preparation and separation runs are essential in order to obtain reproducible separations . Expansion of these types of rapid, efficient microbial separations could have profound effects on many branches of science and technology. Microbes Infect, 1999 Oct, 1(12), 1005 - 14 Pathogenicity of microbes associated with cystic fibrosis; Hutchison ML et al.; Cystic fibrosis patients are exceptionally prone to colonisation by a narrow spectrum of pathogenic bacteria . Since pulmonary infection presently, and for the foreseeable future, plays such a major role in CF lung disease, we review the microbes that are classically associated with CF and the virulence, inflammatory potential and resistance mechanisms which contribute to the reduction in life expectancy for colonised CF patients. Trends Cell Biol, 1999 Dec, 9(12), M2 - 5 Millennium bugs; Davies J; Microbiology has a long way to go . Microbes are ubiquitous, and all other life forms in the biosphere exist solely because of them, but, as less than 1% of microorganisms can be grown in the laboratory, more than a century of research has revealed only the tip of the iceberg concerning this most crucial of life sciences . There are many intellectual challenges remaining . The flow of complete sequences of bacterial genomes is likely to spawn renewed research in answering many questions of concern to academic, medical and industrial interests . Elucidating the roles of microbes, the oldest and most vital inhabitants of the biosphere, in the evolutionary process and in the maintenance of other life forms will be the major thrust in the years to come. Can J Gastroenterol, 1999 Sep, 13(7), 563 - 6 Helicobacter pylori-epithelial cell interactions: from adhesion to apoptosis; Jones NL et al.; Studies suggest that host cell signal transduction cascades are manipulated during infection with microbes, including the gastric pathogen Helicobacter pylori . Several putative adhesins have been proposed to mediate the attachment of H pylori to gastric epithelial cells . Following bacterial binding, a series of signalling pathways are activated in the infected gastric epithelial cell . These signals include both cytoplasmic (such as vacuolization, tyrosine phosphorylation and elevation of cytosolic calcium) and nuclear (proliferation, apoptosis and chemokine transcription) events . Research aimed at elucidating the interactions that occur between the host cell and the bacterium during infection should improve the limited knowledge of disease pathogenesis. J Neurovirol, 1999 Dec, 5(6), 591 - 603 Molecular and cellular mechanisms for microbial entry into the CNS; Zhang JR et al.; A number of pathogenic microbes including neuroinvasive viruses, bacteria and parasites are capable of entry into the central nervous system (CNS) and cause a variety of clinical manifestations . The cellular and molecular mechanisms for the CNS invasion have been extensively studied in the last two decades . Viruses invade neurons and thereby cause encephalitis or peripheral neuritis, while bacteria enter the cerebrospinal fluid (CSF) and cause meningitis . In contrast, the mechanisms for parasitic neuroinvasion are much more complex and less clear . The capabilities that enable these elite subsets of pathogens to engineer uptake into the CNS will be the subject of this review. J Agric Food Chem, 1999 May, 47(5), 2082 - 7 Changes in volatile flavor components of guava juice with high-pressure treatment and heat processing and during storage; Yen GC et al.; The changes in volatile flavor components of guava juice during pressure processing (25 degrees C, 600 MPa, 15 min), heat processing (95 degrees C, 5 min), and storage at 4 and 25 degrees C were evaluated by purge and trap/gas chromatography/mass spectrometry . Esters were the major volatile fraction in guava juice, and alcohols were the second . Pressure processing could maintain the original flavor distribution of the juice . Heat processing (95 degrees C, 5 min) caused decreases in the majority of flavor components in the juice when compared with freshly extracted juice . High-pressure treatment at 600 MPa for 15 min can effectively sterilize microbes but partially inactivate enzymes of guava juice; therefore, volatile components in pressure-treated juice gradually changed during storage periods . Pressure-treated guava juice showed increases in methanol, ethanol, and 2-ethylfuran with decreases in the other components during storage period . Nevertheless, the volatile distribution of 600 MPa treated guava juice was similar to that of freshly extracted juice when stored at 4 degrees C for 30 days. Graefes Arch Clin Exp Ophthalmol, 1999 Nov, 237(11), 915 - 9 Corneal biopsy in keratitis performed with the microtrephine; Kompa S et al.; BACKGROUND: The aetiology of most cases of keratitis remains unclear because the causative agents respond to broad-spectrum antibiotics . Problems occur when they become resistant to local therapies . Further diagnostic measures such as corneal scrapings or biopsies are then necessary . In order to ensure early and gentle biopsy followed by effective diagnosis within 24 h, corneal biopsy specimens were obtained with a microtrephine . PATIENTS AND METHODS: Microbiopsies were obtained from 28 patients suffering from corneal infiltrates or ulcerative keratitis . Different stainings were used to identify the pathogens . Photographs of the clinical healing process were taken immediately after biopsy and during the follow-up . RESULTS: One hundred and ten microbiopsies were performed . One hundred and eighteen specimens could be obtained . No perforation occurred . In 5 of 10 cases in which herpetic keratitis was predicted, herpes DNA could be confirmed . The other five cases were found to be caused by other microbes . In 15 of 18 cases, the bacterial pathogen could be confirmed by Gram's stain diagnosis after microtrephination . Corneal smear was positive in only 7 of these cases . In 2 of 6 cases, predicted to be caused by fungi, lactophenol-blue staining of the microbiopsies showed positive results . Corneal smear was positive in only 1 of these 2 fungal cases . No intraoperative or postoperative complications occurred . No worsening of the disease as a result of treatment could be observed . CONCLUSIONS: The confirmation of microbial cause of keratitis is more effective using microbiopsy than with corneal smears . As a result of the effective treatment after biopsy diagnosis, the majority of cases of keratitis healed . Local therapy seems to have been optimised due to the unroofing of infection during biopsy as well . Therefore microbiopsy in combination with laboratory diagnosis may prove to be a very useful diagnostic and possibly therapeutic method in the clinical routine. Zhonghua Min Guo Wei Sheng Wu Ji Mian Yi Xue Za Zhi, 1996 Nov, 29(4), 185 - 96 Microbial corrosion of aluminum alloy; Yang SS et al.; Several microbes were isolated from the contaminated fuel-oil in Taiwan and the microbial corrosion of aluminum alloy A356-T6 was tested by MIL-STD-810E test method . Penicillium sp . AM-F5 and Cladosporium resinac ATCC 22712 had significant adsorption and pitting on the surface of aluminum alloy, Pseudomonas acruginosa AM-B5 had weak adsorption and some precipitation in the bottom, and Candida sp . AM-Y1 had the less adsorption and few cavities formation on the surface . pH of the aqueous phase decreased 0.3 to 0.7 unit for 4 months of incubation . The corrosion of aluminum alloy was very significant in the cultures of Penicillium sp . AM-F2, Penicillium sp . AM-F5 and C . resinac ATCC 22712 . The major metabolites in the aqueous phase with the inoculation of C . resinac were citric acid and oxalic acid, while succinic acid and fumaric acid were the minors. Science, 1999 Dec 10, 286(5447), 2141 - 4 Geomicrobiology of subglacial ice above Lake Vostok, Antarctica; Priscu JC et al.; Data from ice 3590 meters below Vostok Station indicate that the ice was accreted from liquid water associated with Lake Vostok . Microbes were observed at concentrations ranging from 2.8 x 10(3) to 3.6 x 10(4) cells per milliliter; no biological incorporation of selected organic substrates or bicarbonate was detected . Bacterial 16S ribosomal DNA genes revealed low diversity in the gene population . The phylotypes were closely related to extant members of the alpha- and beta-Proteobacteria and the Actinomycetes . Extrapolation of the data from accretion ice to Lake Vostok implies that Lake Vostok may support a microbial population, despite more than 10(6) years of isolation from the atmosphere. Am J Respir Crit Care Med, 1999 Dec, 160(6), 1943 - 6 Nitric oxide and proinflammatory cytokines in nasal lavage fluid associated with symptoms and exposure to moldy building microbes; Hirvonen MR et al.; Epidemiological data indicate that living or working in a moldy building is associated with increased risk of respiratory symptoms and disease related to inflammatory reactions, but biochemical evidence linking cause and effect is still scarce . The staff working in a mold-contaminated school, and a reference group without such exposure, were studied . Nasal lavage was performed and health data were collected with a questionnaire at the end of the spring term, after a 2.5-mo summer vacation, and at the end of the fall term . Here we show that concentrations of tumor necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6), and nitric oxide (NO) in nasal lavage fluid were significantly higher in the exposed than in the control subjects at the end of the first exposure period . These inflammatory mediators decreased to reference group concentrations during the period when there was no exposure and the production of NO and IL-6 increased again during the reexposure in the fall term . Reports of cough, phlegm, rhinitis, eye irritation, and fatigue paralleled the changes in the measured inflammatory markers . These results point to an association between inflammatory markers in the nasal lavage fluid, the high prevalence of respiratory symptoms among the occupants, and chronic exposure to molds in the indoor environment. Int J Cancer, 2000 Jan 1, 85(1), 35 - 9 Chlamydia trachomatis infection as a risk factor for invasive cervical cancer; Koskela P et al.; Cervical carcinoma is a sexually transmitted disease most strongly linked with human-papillomavirus (HPV) infection . We conducted a prospective sero-epidemiologic study to evaluate the role of Chlamydia trachomatis infection in the development of cervical carcinoma, with invasive cancer as an end point . A nested case-control study within a cohort of 530000 Nordic women was performed . Linking data files of 3 Nordic serum banks and the cancer registries of Finland, Norway and Sweden identified 182 women with invasive cervical carcinoma diagnosed during a mean follow-up of 5 years after serum sampling . The serum samples of the cases and matched cancer-free controls were analyzed for IgG antibodies to C . trachomatis, C . pneumoniae (a control microbe) and HPV types 16, 18 and 33, as well as for serum cotinine (an indicator of tobacco smoking) . Serum antibodies to C . trachomatis were associated with an increased risk for cervical squamous-cell carcinoma (HPV- and smoking-adjusted OR, 2.2; 95% CI, 1.3-3.5) . The association remained also after adjustment for smoking both in HPV16-seronegative and -seropositive cases (OR, 3.0; 95% CI, 1.8-5.1; OR, 2.3, 95% CI, 0 . 8-7.0 respectively) . No such association was found for C . pneumoniae . Our prospective study provides sero-epidemiologic evidence that infection with C . trachomatis confers an increased risk for subsequent development of invasive squamous-cell carcinoma of the uterine cervix . Appl Environ Microbiol, 1999 Dec, 65(12), 5314 - 21 A study of deep-sea natural microbial populations and barophilic pure cultures using a high-pressure chemostat; Wirsen CO et al.; Continuous cultures in which a high-pressure chemostat was used were employed to study the growth responses of (i) deep-sea microbial populations with the naturally occurring carbon available in seawater and with limiting concentrations of supplemental organic substrates and (ii) pure cultures of copiotrophic barophilic and barotolerant deep-sea isolates in the presence of limiting carbon concentrations at various pressures, dilution rates, and temperatures . We found that the growth rates of natural populations could not be measured or were extremely low (e.g., a doubling time of 629 h), as determined from the difference between the dilution rate and the washout rate . A low concentration of supplemental carbon (0.33 mg/liter) resulted in positive growth responses in the natural population, which resulted in an increase in the number of cells and eventually a steady population of cells . We found that the growth responses to imposed growth pressure by barophilic and barotolerant pure-culture isolates that were previously isolated and characterized under high-nutrient-concentration conditions were maintained under the low-nutrient-concentration limiting conditions (0.33 to 3.33 mg of C per liter) characteristic of the deep-sea environment . Our results indicate that deep-sea microbes can respond to small changes in substrate availability . Also, barophilic microbes that are copiotrophic as determined by their isolation in the presence of high carbon concentrations and their preference for high carbon concentrations are versatile and are able to compete and grow as barophiles in the low-carbon-concentration oligotrophic deep-sea environment in which they normally exist. Appl Environ Microbiol, 1999 Dec, 65(12), 5234 - 41 Ubiquity and diversity of dissimilatory (per)chlorate-reducing bacteria; Coates JD et al.; Environmental contamination with compounds containing oxyanions of chlorine, such as perchlorate or chlorate {(per)chlorate} or chlorine dioxide, has been a constantly growing problem over the last 100 years . Although the fact that microbes reduce these compounds has been recognized for more than 50 years, only six organisms which can obtain energy for growth by this metabolic process have been described . As part of a study to investigate the diversity and ubiquity of microorganisms involved in the microbial reduction of (per)chlorate, we enumerated the (per)chlorate-reducing bacteria (ClRB) in very diverse environments, including pristine and hydrocarbon-contaminated soils, aquatic sediments, paper mill waste sludges, and farm animal waste lagoons . In all of the environments tested, the acetate-oxidizing ClRB represented a significant population, whose size ranged from 2.31 x 10(3) to 2.4 x 10(6) cells per g of sample . In addition, we isolated 13 ClRB from these environments . All of these organisms could grow anaerobically by coupling complete oxidation of acetate to reduction of (per)chlorate . Chloride was the sole end product of this reductive metabolism . All of the isolates could also use oxygen as a sole electron acceptor, and most, but not all, could use nitrate . The alternative electron donors included simple volatile fatty acids, such as propionate, butyrate, or valerate, as well as simple organic acids, such as lactate or pyruvate . Oxidized-minus-reduced difference spectra of washed whole-cell suspensions of the isolates had absorbance maxima close to 425, 525, and 550 nm, which are characteristic of type c cytochromes . In addition, washed cell suspensions of all of the ClRB isolates could dismutate chlorite, an intermediate in the reductive metabolism of (per)chlorate, into chloride and molecular oxygen . Chlorite dismutation was a result of the activity of a single enzyme which in pure form had a specific activity of approximately 1,928 micromol of chlorite per mg of protein per min . Analyses of the 16S ribosomal DNA sequences of the organisms indicated that they all belonged to the alpha, beta, or gamma subclass of the Proteobacteria . Several were closely related to members of previously described genera that are not recognized for the ability to reduce (per)chlorate, such as the genera Pseudomonas and Azospirllum . However, many were not closely related to any previously described organism and represented new genera within the Proteobacteria . The results of this study significantly increase the limited number of microbial isolates that are known to be capable of dissimilatory (per)chlorate reduction and demonstrate the hitherto unrecognized phylogenetic diversity and ubiquity of the microorganisms that exhibit this type of metabolism. Radiology, 1999 Oct, 213(1), 299 - 300 US probes: risk of cross infection and ways to reduce it--comparison of cleaning methods; Fowler C et al.; After their use at ultrasonography (US) in the intensive therapy unit, probes were used to directly inoculate blood agar plates before and after various cleaning procedures . The uncleaned probes transmitted large numbers of clinically important microbes . Simple cleaning methods were effective in reducing transmission among certain patients: fit patients, double paper wipe; patients at risk of contracting infection, single paper wipe followed by alcohol wipe; patients with a potential source of infection, single paper wipe followed by alcohol wipe. Plant J, 1999 Nov, 20(3), 371 - 8 Technical advance: single pollen typing combined with laser-mediated manipulation; Matsunaga S et al.; We combined single pollen typing with laser-mediated manipulation . After drilling a hole in the wall of a pollen grain from a dioecious plant (Silene latifolia) with a UV-laser microbeam, the single pollen grain was recovered directly in the cap of a PCR tube, using a non-contact method called laser pressure catapulting . The entire genome of the single pollen grain was then amplified with improved primer-extension-preamplification PCR (I-PEP PCR) . Nested PCR with sequence tagged site (STS)-specific primers was used to analyze several loci in the haploid genome . The single copy gene MROS1 was detected in most of the single pollen grains analyzed . Bgl10, which is localized on the Y chromosome, was detected in approximately half of the pollen grains . MROS3 is reported to be localized on the X chromosome . Using inverse PCR, we isolated two genomic clones of MROS3: MROS3A and MROS3B . The single pollen analysis using nested PCR showed that MROS3A and MROS3B are derived from different loci that are not located on the X chromosome . Single pollen typing not only reveals sex chromosome-linkage within the haploid genome, but can also discriminate between alleles and different loci . This method should also be useful for measuring recombination frequencies without genetic crossover analysis. Int J Mol Med, 1999 Dec, 4(6), 655 - 8 Brucellosis outbreak: analysis of risk factors and serologic screening; Mishal J et al.; Israel is one of the Mediterranean countries in which Brucellosis is endemic . As recently there has been a Brucellosis outbreak in a kibbutz, the aim of this study is to identify asymptomatic infected Kibbutz members, and to delineate the manner of infection in this setting . Therefore, all the asymptomatic Kibbutz members were screened by the Rose Bengal test for Brucellosis, while both patients and healthy members were asked to fill in a questionnaire in order to pinpoint the manner of infection, and signs and symptoms of the disease . In addition to the 14 patients with Brucellosis, 2 other Kibbutz members were also found to be infected by the screening tests . Analysis of the data of the questionnaires from 142 healthy and 16 patients disclosed that almost all of the infected patients (15/16) worked in the cowshed, as opposed to only 24 out of 142 (16.9%) of the healthy members . The infected tended to participate more in calf deliveries, and had contact with cow's blood and placenta, compared with the healthy subjects (P<0.001), while there were no significant differences with respect to having cuts on hands, or working in the cowshed without gloves . In addition, 15 out of 16 (93.8%) infected persons also drank unpasteurized milk, as compared with only 17 of the 142 (12%) healthy members (P<0.001), and thus were exposed to 2 major risk factors (working in the cowshed and consumption of unpasteurized milk) . As the cows of the Kibbutz's cowshed were affected by Brucella melitensis (which usually affects flocks of goats and sheep rather than cows), the microbe was probably transmitted to the cowshed from neighboring flocks by wandering dogs, and then to the infected humans. Immunol Rev, 1999 Aug, 170, 65 - 72 Regulation of the IL-12/IL-12R axis: a critical step in T-helper cell differentiation and effector function; Sinigaglia F et al.; Interleukin (IL)-12 is required for the development of T-helper (Th) 1 cells, which have been shown to be important for protective cell-mediated immune responses against a variety of intracellular pathogens . Recent studies have clarified the sources and the regulation of IL-12 production leading to Th1 development against microbes . Expression of IL-12R is necessary for maintaining IL-12 responsiveness and controlling Th1 lineage commitment . Advances in this area have included a broader understanding of the factors involved in the regulation of the IL-12R beta 2 signaling component . Expression of this receptor subunit in humans is critically influenced by IL-12 and type I interferons . IL-12 signaling results in STAT4 activation and interferon (IFN)-gamma production . Recent evidence suggests that IL-12 also modulates a number of genes involved in leukocyte trafficking . Thus, IL-12 is not only an important proinflammatory cytokine, which induces production of IFN-gamma and subsequent activation of phagocytic cells but also plays a major role in regulating the migration and proper positioning of effector cells. Immunol Rev, 1999 Aug, 170, 7 - 27 Personal historical chronicle of six decades of basic and applied research in virology, immunology, and vaccinology; Hilleman MR; The sciences of vaccinology and of immunology were created just two centuries ago by Jenner's studies of prevention of smallpox by inoculation with cowpox virus . This rudimentary beginning was expanded greatly by the giants of late 19th and early 20th centuries biomedical sciences . The period from 1930 to 1950 was a transitional era with the introduction of chick embryos and minced tissues for propagating viruses and rickettsiae in vitro for vaccines . Modern era vaccinology began about 1950 as a continuum of notable advances made during the 1940s and World War II . Present vaccinology is based largely on breakthroughs in cell culture, bacterial polysaccharide chemistry, molecular biology, and immunology . By invitation, the author, who is a microbe hunter in fact, was asked to chronicle his six decades of pioneering achievements in basic and applied virology, bacteriology, immunology, molecular biology, epidemiology, and cancer, with special reference to the pioneering creation of most of the present day vaccines . Knowledge of the past may guide the present and future . This chronicle will have achieved its legacy if it helps others to understand the why and how of the past that may help to create the substance of the future. Br J Gen Pract, 1999 Jun, 49(443), 436 - 40 Influence of prescription patterns in general practice on anti-microbial resistance in Norway; Lindbaek M et al.; BACKGROUND: The global pandemic of antibiotic resistance is causing considerable concern, and a major reason for the growing world-wide resistance problem is the overuse of anti-infective drugs, especially the use of broad spectrum antibiotics . This is still a relatively minor problem in most of the Nordic countries where the consumption of antibiotics is less than half of that reported from southern Europe and the United States of America (USA) . AIM: To describe the resistance pattern among common respiratory tract pathogens in Norwegian general practice, the national consumption of antibiotics, and GPs' prescription patterns for respiratory tract infections . To offer some suggestions as to why Norway has maintained a favourable situation regarding resistant microbes during the past 10 years . METHODS: An analysis of the prescription patterns in Norwegian general practice . RESULTS: There is a low total prescription volume of antibiotics compared with other countries . Penicillin V is the most commonly used antibiotic for the most common airway diseases in general practice in Norway . CONCLUSION: Although there is a low prevalence of antibiotic resistance in Norway, there is still a great potential for reducing the unnecessary antibiotic prescribing for the most common respiratory illnesses. Am J Hypertens, 1999 Oct, 12(1O Pt 1), 1030 - 7 Comparisons of the effects of different long-acting delivery systems on the pharmacokinetics and pharmacodynamics of diltiazem; Smith DH et al.; The benzothiazepine calcium channel antagonist diltiazem is a short-acting drug . To achieve effective 24-h blood pressure control with once-daily dosing, it relies on various extended drug-delivery systems that have grown in importance as a result of the recent reports relating the use of short-acting calcium channel antagonists to increased cardiovascular morbidity . This study examines the pharmacokinetics and resulting pharmacodynamics of two different delivery systems, each loaded with 240 mg of diltiazem and administered to 40 moderately hypertensive patients in a randomized, double-blind crossover trial . After a 4-week, single-blind placebo lead-in, patients with a clinical diastolic blood pressure of > or =100 mm Hg were randomized to either the single or dual microbead diltiazem delivery system for a 4-week period . At the end of this period, each subject was evaluated with 24-h ambulatory blood pressure monitoring and subjected to 24-h inpatient pharmacokinetic analysis on separate days . This was followed by a similar 4-week period in which each subject was treated with the alternative delivery system . For diltiazem, the area under the curve for plasma concentration versus time and the maximum plasma concentration attained by the single microbead system exceeded the values achieved by the dual bead system by 15% and 25%, respectively . These differences were greatest from the 3rd through the 13th h after dosing . During this period, both systolic and diastolic ambulatory blood pressure was significantly lower when the single microbead system was used . When compared with baseline blood pressure, blood pressure reductions achieved with the single microbead system exceeded reductions achieved with the dual microbead system by at least 2 mm Hg for 10 of the 24 postdose hours . Heart rates were slightly reduced but not significantly different . This improved blood pressure control at higher plasma levels of diltiazem suggests that a more efficient delivery system could provide better blood pressure control for identical doses of diltiazem. Vaccine, 1999 Oct 29, 17 Suppl 3, S64 - 7 Paul Ehrlich and O.T . Avery: pathfinders in the search for immunity; Krause RM; This paper is concerned with the use of passive immunity for the prevention and treatment of infections from a historic perspective, particularly in regard to the research of Paul Ehrlich on diphtheria and that of O.T . Avery on pneumonia . It is timely to reexamine this matter, particularly in regard to virus infections, in view of the difficulties in developing antiviral drugs . In addition, specific antibodies can serve as an alternative therapy for antibiotic resistant microbes.It is worth recalling in the history of vaccine development that success in passive immunity has been the key element in devising a successful strategy to develop a vaccine to produce humoral immunity. J Immunol, 1999 Nov 15, 163(10), 5519 - 25 Defects in the generation of IFN-gamma are overcome to control infection with Leishmania donovani in CC chemokine receptor (CCR) 5-, macrophage inflammatory protein-1 alpha-, or CCR2-deficient mice; Sato N et al.; We investigated the immune responses in mice lacking CCR2, CCR5, or macrophage inflammatory protein-1 alpha (MIP-1 alpha), a ligand for CCR5, in two situations: following T cell stimulation or after challenge with Leishmania donovani, an intracellular microbe whose control is dependent on a Th1 immune response . Mice deficient in CCR5, MIP-1 alpha, or CCR2 had reduced IFN-gamma responses following ligation of the TCR . Reduced IFN-gamma responses following PMA and ionomycin were also observed in CD8+ T cells of CCR5-/- and CCR2-/- mice . During the early phases of infection, all three knockout mice had low Ag-specific IFN-gamma responses . However, this reduced IFN-gamma response was overcome during a state of persistent Ag stimulation (chronic infection), and was not associated with an adverse parasitologic outcome in any of the gene-targeted mouse strains . To the contrary, during the late phase of infection, an exaggerated Ag-specific IFN-gamma response was evident in CCR5-/- and MIP-1 alpha-/- mice, and this correlated with an enhanced control of parasite replication . Although granuloma formation was abnormal in each of the knockout mice, there was no correlation between the number or architecture of the granulomas and parasite burden . Collectively, these findings indicate an important role for CCR5, MIP-1 alpha, and CCR2 in granulomatous inflammation, and that CCR5 and MIP-1 alpha, possibly acting through CCR5, might play a deleterious role in the outcome of chronic L . donovani infection . Our data also suggest that there might be cross-talk between TCR and chemokine receptor signaling pathways. Int J Syst Bacteriol, 1999 Oct, 49 Pt 4, 1809 - 16 A novel species of thermoacidophilic archaeon, Sulfolobus yangmingensis sp . nov; Jan RL et al.; A novel microbe was isolated from a geothermal vent in Yang-Ming National Park in northern Taiwan . This spherical microbe with mean cell diameter of 1.1 +/- 0.2 microns is a facultatively chemolithoautotrophic archaeon that grows on elemental sulfur and reduced sulfur compounds . The optimal pH and temperature for growth are 4.0 (pH range 2.0-6.0) and 80 degrees C (temperature range 65-95 degrees C) . Its membranes contain the lipids calditoglycerocaldarchaeol and caldarchaeol, which are common to other members of the Sulfolobaceae . Like Sulfolobus acidocaldarius, Sulfolobus shibatae and Sulfolobus solfataricus, the new isolate utilizes sugars and amino acids effectively as sole carbon sources . The G + C content of the genomic DNA was 42 mol% . DNA of the isolate hybridized weakly to the DNA of other Sulfolobus species . Phylogenetic analysis of the 16S rRNA indicated that the new isolate represents a deep branch within the genus Sulfolobus . On the basis of these properties, the new isolate appears to represent a new species of Sulfolobus, for which the name Sulfolobus yangmingensis sp . nov . is proposed . The type strain is strain YM1T. Naturwissenschaften, 1999 Nov, 86(11), 507 - 15 Modeling plasticity at the micrometer scale Gao H, Huang Y, Nix WD. Plastic deformation exhibits a strong dependence upon size at and below micrometer length scales . To model micrometer scale plasticity we propose a theory called mechanism-based strain gradient plasticity (MSG), which connects microscale dislocation interactions to mesoscale plasticity via a hierarchical framework linking Taylor's dislocation hardening model to strain gradient plasticity . We distinguish the microscale, at which dislocation interaction is considered, from the mesoscale, at which the plasticity theory is formulated . The MSG theory differs from all existing phenomenological theories of plasticity in its mechanism-based guiding principles . We show that the MSG theory gives predictions in agreement with microindentation, microtorsion, and microbending experiments. J Oral Pathol Med, 1999 Nov, 28(10), 451 - 5 Crystalloid architecture of a sialolith in a minor salivary gland; Riesco JM et al.; A sialolith from a minor salivary gland of the mucosa of the upper lip was studied morphologically and analytically . Under stereoscopic microscopic visualisation, no core or any concentric laminar structure was found in the sialolith and it had a transparent glassy appearance . Scanning electron microscopy revealed differences between the internal structure of the sialolith and its external structure . No microbes were observed but some mineralized inclusion bodies were seen . X-ray diffraction indicated the absence of inorganic crystals in the sialolith, while energy dispersive x-ray microanalysis revealed a high content of S followed by Ca, Si and Na and little and scarce P on the fracture surface, with S, Na, Ca and P being distributed throughout the external coating of the sialolith . The results suggest that the sialolith was young and consisted of a crystalloid body with an incipient coating undergoing calcification. Clin Diagn Lab Immunol, 1999 Nov, 6(6), 970 - 6 Sterols of Pneumocystis carinii hominis organisms isolated from human lungs; Kaneshiro ES et al.; The opportunistic pathogen Pneumocystis carinii causes pneumonia (P . carinii pneumonia, or PCP) in immunocompromised individuals such as AIDS patients . Rat-derived P . carinii carinii organisms have distinct sterols which are not synthesized by mammals and not found in other microbes infecting mammalian lungs . The dominant sterol present in the organism is cholesterol (which is believed to be scavenged from the host), but other sterols in P . carinii carinii have an alkyl group at C-24 of the sterol side chain (C(28) and C(29) 24-alkylsterols) and a double bond at C-7 of the nucleus . Recently, pneumocysterol (C(32)), which is essentially lanosterol with a C-24 ethylidene group, was detected in lipids extracted from a formalin-fixed human P . carinii-infected lung, and its structures were elucidated by gas-liquid chromatography, mass spectrometry, and nuclear magnetic resonance spectrometry in conjunction with analyses of chemically synthesized authentic standards . The sterol composition of isolated P . carinii hominis organisms has yet to be reported . If P . carinii from animal models is to be used for identifying potential drug targets and for developing chemotherapeutic approaches to clear human infections, it is important to determine whether the 24-alkylsterols of organisms found in rats are also present in organisms in humans . In the present study, sterol analyses of P . carinii hominis organisms isolated from cryopreserved human P . carinii-infected lungs and from bronchoalveolar lavage fluid were performed . Several of the same distinct sterols (e.g., fungisterol and methylcholest-7-ene-3beta-ol) previously identified in P . carinii carinii were also present in organisms isolated from human specimens . Pneumocysterol was detected in only some of the samples. Annu Rev Microbiol, 1999, 53, 103 - 28 Aerotaxis and other energy-sensing behavior in bacteria; Taylor BL et al.; Energy taxis is widespread in motile bacteria and in some species is the only known behavioral response . The bacteria monitor their cellular energy levels and respond to a decrease in energy by swimming to a microenvironment that reenergizes the cells . This is in contrast to classical Escherichia coli chemotaxis in which sensing of stimuli is independent of cellular metabolism . Energy taxis encompasses aerotaxis (taxis to oxygen), phototaxis, redox taxis, taxis to alternative electron acceptors, and chemotaxis to a carbon source . All of these responses share a common signal transduction pathway . An environmental stimulus, such as oxygen concentration or light intensity, modulates the flow of reducing equivalents through the electron transport system . A transducer senses the change in electron transport, or possibly a related parameter such as proton motive force, and initiates a signal that alters the direction of swimming . The Aer and Tsr proteins in E . coli are newly recognized transducers for energy taxis . Aer is homologous to E . coli chemoreceptors but unique in having a PAS domain and a flavin-adenine dinucleotide cofactor that is postulated to interact with a component of the electron transport system . PAS domains are energy-sensing modules that are found in proteins from archaea to humans . Tsr, the serine chemoreceptor, is an independent transducer for energy taxis, but its sensory mechanism is unknown . Energy taxis has a significant ecological role in vertical stratification of microorganisms in microbial mats and water columns . It plays a central role in the behavior of magnetotactic bacteria and also appears to be important in plant-microbe interactions. Anal Chem, 1999 Oct 15, 71(20), 4727 - 33 Analysis for TNF-alpha using solid-phase affinity capture with radiolabel and MALDI-MS detection; Hurst GB et al.; Screening of mutant mice for subtle phenotypes requires sensitive, high-throughput analyses of sentinel proteins in functional pathways . The cytokine TNF-alpha is upregulated during inflammatory reactions associated with autoimmune diseases . We have developed a method to monitor the concentration of TNF-alpha under physiological conditions . TNF-alpha is captured, purified, and concentrated using monoclonal antibody-coated microbeads . The capture is efficient (> 80%) and can be used in the concentration range < 100 pg/mL to > 50 ng/mL, as determined by detection of 125I-labeled TNF-alpha . The bead capture of TNF-alpha can be combined with direct detection by MALDI-MS for sample concentrations of > 10 ng/mL . TNF-alpha can be captured and detected from diluted mouse serum, with minimal interferences observed in the MALDI spectrum . This method is adaptable to high-throughput sample handling with microfluidic devices and automated mass spectrometric analysis. J Soc Biol, 1999, 193(3), 307 - 9 {Perspectives and hopes in infectious pathology}; Ramuz M; Over the last few decades, changes in socio-economic conditions and social practices as well as aggressive therapy of many diseases have led to the emergence of new infectious pathologies . These new pathologies are either associated with newly identified microbial species or the emergence of known microbes which have encountered new environments in which they are able to cause disease . Recent progress has allowed us to understand the mechanisms by which these pathogens express their virulence and will certainly allow us to diagnose and treat these infections more efficiently in the future. Proc Natl Acad Sci U S A, 1999 Oct 26, 96(22), 12275 - 80 Identification of lumichrome as a sinorhizobium enhancer of alfalfa root respiration and shoot growth Phillips DA, Joseph CM, Yang GP, Martinez-Romero E, Sanborn JR, Volpin H. Sinorhizobium meliloti bacteria produce a signal molecule that enhances root respiration in alfalfa (Medicago sativa L.) and also triggers a compensatory increase in whole-plant net carbon assimilation . Nuclear magnetic resonance, mass spectrometry, and ultraviolet-visible absorption identify the enhancer as lumichrome, a common breakdown product of riboflavin . Treating alfalfa roots with 3 nM lumichrome increased root respiration 21% (P < 0.05) within 48 h . A closely linked increase in net carbon assimilation by the shoot compensated for the enhanced root respiration . For example, applying 5 nM lumichrome to young alfalfa roots increased plant growth by 8% (P < 0.05) after 12 days . Soaking alfalfa seeds in 5 nM lumichrome before germination increased growth by 18% (P < 0.01) over the same period . In both cases, significant growth enhancement (P < 0.05) was evident only in the shoot . S . meliloti requires exogenous CO(2) for growth and may benefit directly from the enhanced root respiration that is triggered by lumichrome . Thus Sinorhizobium-alfalfa associations, which ultimately form symbiotic N(2)-reducing root nodules, may be favored at an early developmental stage by lumichrome, a previously unrecognized mutualistic signal . The rapid degradation of riboflavin to lumichrome under many physiological conditions and the prevalence of riboflavin release by rhizosphere bacteria suggest that events demonstrated here in the S . meliloti-alfalfa association may be widely important across many plant-microbe interactions. Sci Total Environ, 1999 Sep 15, 236(1-3), 173 - 80 Influence of microbes on the mobilization, toxicity and biomethylation of arsenic in soil; Turpeinen R et al.; To understand the effects of microbial activity on the mobilization and speciation of arsenic in soil, the cycling of arsenic was studied in microcosm experiments under laboratory conditions . Particular attention was paid to the biomethylation of arsenic and to the toxicity of inorganic and organic arsenic species for microbes . Microbes enhanced mobilization of arsenic from soil by 19-24% compared to formaldehyde inhibited controls . Formation of dissolved methylated arsenic species by microbes was low (< 0.1%) during the 5-day incubation . Even though methylation may function as a detoxification method, it was of minor importance in the soil tested. J Am Acad Dermatol, 1999 Nov, 41(5 Pt 1), 733 - 7 Bullous erysipelas: A retrospective study of 26 patients; Guberman D et al.; BACKGROUND: Erysipelas is a superficial form of cellulitis caused by a variety of microbes, and it responds to antibiotic treatment . During the past few years we treated several patients with a bullous form of erysipelas involving the lower legs . We believe their disease had a more protracted course than patients with nonbullous erysipelas . OBJECTIVE: We studied bullous erysipelas by conducting a retrospective analysis of 26 patients with bullous erysipelas of the legs treated by the authors during a 5-year period . METHODS: We conducted a retrospective review of the records of all patients with a diagnosis of bullous erysipelas who were treated at the Department of Dermatology, Hadassah Medical Center, Jerusalem, between the years 1992 and 1996 . Data regarding patients with nonbullous erysipelas were obtained from the medical center's computerized data pool . RESULTS: A total of 26 cases of bullous erysipelas were found, comprising 22 women and 4 men whose ages ranged from 28 to 87 (mean, 58.8) years . The average hospital stay was 20.57 days (range, 12 to 46 days) . The average hospital stay for patients with nonbullous erysipelas and cellulitis treated in the same department by the authors during the study period was 10.6 days (range, 2 to 54 days) . CONCLUSION: Bulla formation is a complication of erysipelas, seen in our series in 5.2% of the patients (26 of 498 admissions for erysipelas and cellulitis) . The course of the disease is protracted, requiring longer medical attention. Drug Dev Ind Pharm, 1999 Oct, 25(10), 1129 - 35 Investigating the fundamental effects of binders on pharmaceutical tablet performance; Joneja SK et al.; For solid dosage forms, a better understanding of the fundamental properties of the binders helps in developing better formulations and products . The objective of this study was to determine the effects of binder toughness and plastic flow on tablet hardness, friability, and capping . The characteristic of binder toughness was determined, and the correlation between the ejection force of the tablet and the toughness of the binder was established . Evaluation was conducted using acetaminophen tablets with different kinds of binders (i.e., hydroxypropylcellulose, methylcellulose {MC}, povidone {PVP}, starch, etc.) . A rotary tablet press was used for tableting at three different speeds . The properties of binders and acetaminophen tablets were determined using a diametral compression test . The toughness was measured as the curve of the area under the load versus deflection . The microbehavior of these binders was also studied . The acetaminophen tablets with the binders were subjected to predetermined loads and then examined under a scanning electron microscope . The tablets that contained hydroxypropylcellulose as a binder showed the highest toughness and had the lowest ejection force . The ejection force of tablets decreased with increasing concentrations of hydroxypropylcellulose in the dosage forms . The tablets that contained other binders failed by capping and random cracking in the middle . These results show that hydroxypropylcellulose, a thermoplastic polymer, provides the best physical characteristics for the tablets . This effect could help in improving tablet manufacturing conditions (e.g., compression force and speed). Methods, 1999 Sep, 19(1), 8 - 16 Effects of xenobiotics on macrophage function: evaluation in vitro; Hubbard AK; Macrophages participate in a variety of inflammatory and immunologic functions (e.g., phagocytosis, cytokine production, killing of microbes and tumor cells, and processing and presentation of antigen to T lymphocytes) . Because these cells are widely distributed in the circulation and throughout tissue, the effects of xenobiotics on macrophage function can be significant . Measures in vitro of altered function elicited by xenobiotic exposure can include changes in expression of cell surface proteins, in production of oxygen and nitrogen free radicals, in production of cytokines {interleukin (IL)-1, IL-6, IL-12, tumor necrosis factor alpha (TNFalpha)}, in expression of adhesion molecules (ICAM-1), in phagocytosis and intracellular killing, and in antigen presentation . Proc Assoc Am Physicians, 1999 Sep-Oct, 111(5), 373 - 82 The NADPH-dependent oxidase of phagocytes; Nauseef WM; Polymorphonuclear leukocytes (PMNs) represent a prominent cellular element in the innate immune system, serving to ingest exogenous particles and microbes and to kill phagocytosed microorganisms . The microbicidal activity of PMNs depends on the interactions of a broad array of potent systems, including relatively stable degradative proteins as well as labile reactive radicals . These systems can be categorized as oxygen-dependent and nonoxidative mechanisms, although the physiologically relative activity depends on the precisely orchestrated interplay between both systems . The enzyme complex responsible for the activity of the oxygen-dependent system is the respiratory burst oxidase and its important contribution to host defense is best illustrated by the frequent and severe infections seen in individuals whose PMNs lack oxidase activity, namely patients with chronic granulomatous disease (CGD) . Multiple elements comprise the oxygen-dependent system, and significant advances have been made in the past decade in understanding the protein components of the respiratory burst oxidase, their subcellular distribution in resting PMNs, and their agonist-dependent assembly into a functional system at phagosomal and plasma membranes . In parallel, substantial insights into the molecular bases of CGD have likewise been made . Nonetheless there remain significant gaps in our understanding of the precise functional contributions of particular components of the system, the molecular mechanisms that regulate their coordinated assembly, and the role of related proteins in nonphagocytic cells. Clin Chest Med, 1999 Sep, 20(3), 475 - 88, vii Pulmonary host defenses . Implications for therapy; Mason CM et al.; Pulmonary host defenses comprise a redundant system of protective mechanisms against invasion of the lungs by pathogenic microbes . The upper and lower airways are uniquely suited to contain and remove organisms that gain access to the respiratory mucosa . If the balance between host and organism is disputed, however, microbial clearance may be ineffective, and infection established . Pulmonary host defense mechanisms, which provide the basis for several current therapeutic strategies, are reviewed. J Investig Allergol Clin Immunol, 1999 Jul-Aug, 9(4), 248 - 52 Elevated CD154 (CD40 ligand) synthesis in T-cells from allergic patients after nonspecific stimulation in vitro; Markert UR et al.; The interaction of the CD154 molecule (CD40 ligand, gp39) on activated T-cells with the CD40 antigen on B-cells seems to play a key role in immunoglobulin class switching . We aimed to compare the capacity of intracellular CD154 expression after nonspecific stimulation with phorbol-12-myristate-13-acetate and ionomycin on separated T-cells from allergic patients and healthy donors . We analyzed blood from 104 patients allergic to grass pollen, house dust mites or birch pollen, and from 44 healthy donors . Lymphocytes were isolated using a density gradient and B-cells were extracted by magnet-activated cell separation (MACS) using anti-CD19 microbeads . Cells were nonspecifically stimulated for 5 h, permeabilized and stained with anti-CD154 for fluorescence-activated cell sorter analysis . It was found that stimulation induced a 1.4% increase of intracellular CD154+ T-cells; a 4.6% increase of mean channel fluorescence of all T-cells from healthy donors; a 6.1% increase in intracellular CD154+ T-cells; and a 28.1% increase of mean channel fluorescence of all T-cells from allergic patients . The data demonstrated an elevated capability of B-cell independent CD154 synthesis in T-cells from allergic patients when compared to healthy individuals . It is possible that the enhanced IgE production of B-cells from allergic patients might be partly due to the phenomena described. Membr Cell Biol, 1999, 12(6), 805 - 15 Postmitotic reconstruction of nucleoli in culture cells with UV-microbeam photoinactivated centrosome; Neverova AL et al.; Ultraviolet microirradiation of one of the poles of the mitotic spindle of PK cells was performed 1 min after the onset of the anaphase . Formation of the nucleolus in the telophase and G1 period was studied by vital observation, electron microscopy and indirect immunofluorescence using antibodies against B23 protein . Sister cells with nonirradiated centrosomes and cells with partially irradiated cytoplasm were used as controls . During the first hour after the anaphase, the nuclei in both sister cells were identical and contained numerous small dense particles with granular ultrastructure . B23 protein detected in the mitotic poles and at the chromosome surface in the anaphase was dispersed in the cytoplasm in both cells in the early G1 period . Later, control cells did not display any difference from intact cells: nucleoli of a typical structure were formed, B23 protein appeared in the karyoplasm and was then accumulated in the nucleoli and disappeared from the cytoplasm and karyoplasm . Nucleoli in cells with irradiated centrosomes did not achieve the normal size and contained a significantly lower amount of granular component . B23 protein was dispersed in the karyoplasm and was not accumulated in the nucleoli . Nucleoli in cells with irradiated centrosomes contained small dense particles for at least 24 h . Telophase cells where microtubule formation had been inhibited by nocodazole formed normal nucleoli . It shows that the effects observed in cells with irradiated centrosomes are not due to the absence of the microtubule radial system . We conclude that UV microirradiation of the mitotic centrosome disturbs the postmitotic reconstruction of nucleoli probably because of the photodestruction of B23 protein accumulated in the mitotic pole. Trends Biotechnol, 1999 Nov, 17(11), 443 - 7 Automation of functional assays by flow injection fluorescence microscopy; Scampavia LD et al.; Bead-injection spectroscopy is a novel technique that uses immobilized eukaryotic cells on microbeads as a renewable biosensor for fluorescence microscopy . The use of a flow injection instrument allows fast functional assays that generate full kinetic characterization of a drug . Because the cell population is automatically replaced for each assay, variability is minimized, thus allowing greater accuracy. J Ind Microbiol Biotechnol, 1999 Aug, 23(2), 106 - 111 Screening for L-sorbose and L-sorbosone dehydrogenase producing microbes for 2-keto-L-gulonic acid production; Lee HW et al.; Acetic acid bacteria incompletely oxidize L-sorbose to 2-keto-L-gulonic acid (2KLG) by L-sorbose- and L-sorbosone dehydrogenases . In order to isolate novel microorganisms with these enzyme activities, a new screening method has been studied with a presumption that microorganisms reuse their metabolic products when principal carbon sources are exhausted . When various keto-aldonic acid-producing microorganisms were tested for the ability to grow in minimal media containing such products as 2,5-diketo-gluconic acid, 2-keto-D-gluconic acid, 5-keto-D-gluconic acid or 2-keto-L-gulonic acid, they grew with these keto-aldonic acids as the sole carbon source . By enriching the isolates collected from screening samples for their growth in minimal medium containing 2KLG as the sole carbon source, as much as 50% of selected strains showed L-sorbose- and L-sorbosone dehydrogenase activities . In spite of the presence of these enzymes, no significant amount of 2KLG was detected in the culture broth, possibly due to 2KLG reductase activity, indicating that the direct screening for 2KLG producer microorganisms would be less successful . These results suggest that the screening strategy using 2KLG as a carbon source is a useful method for the selective screening of microorganisms with L-sorbose- and L-sorbosone dehydrogenases, and that a similar strategy may be applied to other cases. Appl Environ Microbiol, 1999 Oct, 65(10), 4451 - 7 Pelagic-benthic coupling of nucleic acids in an abyssal location of the northeastern Atlantic Ocean; Dell'Anno A et al.; Spatial and temporal changes in sedimentary nucleic acid concentrations in an abyssal locality of the northeastern Atlantic Ocean were investigated in relation to fluxes of nucleic acids produced in the photic layer . Sediment trap material, collected between 1996 and 1998 at depths of 1,000, 3,000, and 4,700 m, and sediment samples were analyzed for DNA and RNA content . Nucleic acid concentrations in the sediments were very high and displayed significant temporal changes, whereas mesoscale variability was low . DNA and RNA concentrations generally displayed opposite temporal patterns, which are likely to be dependent on the nature and characteristics of DNA and RNA molecules . Nucleic acid fluxes were high and displayed clear seasonal changes apparently coupled with seasonal pulses of primary production . However, while median values of DNA fluxes were relatively similar in all sediment traps, median values of RNA fluxes almost doubled from the 1,000- to the 4,700-m depth, suggesting differ