Ann N Y Acad Sci, 1999 Jun 30, 878, 404 - 12 C-telopeptide pyridinoline cross-links . Sensitive indicators of periodontal tissue destruction; Giannobile WV; C-telopeptides and related pyridinoline cross-links of bone Type I collagen are sensitive markers of bone resorption in osteolytic diseases such as osteoporosis and osteoarthritis . We have studied the release of C-telopeptide pyridinoline crosslinks of Type I collagen as measures of bone destruction in periodontal disease . Studies in preclinical animal models and humans have demonstrated the relationship between radiographic bone loss and crevicular fluid C-telopeptide levels . We have recently found that C-telopeptide levels correlate strongly with microbial pathogens associated with periodontitis and around endosseous dental implants . Host-modulation of bone-related collagen breakdown has been shown by studies in humans demonstrating that MMP inhibition blocks tissue destruction and release of C-telopeptides in patients with active periodontal disease. Arch Microbiol, 1999 Aug, 172(2), 83 - 94 Specific detection of different phylogenetic groups of chemocline bacteria based on PCR and denaturing gradient gel electrophoresis of 16S rRNA gene fragments; Overmann J et al.; Specific amplification of 16S rRNA gene fragments in combination with denaturing gradient gel electrophoresis (DGGE) was used to generate fingerprints of Chromatiaceae, green sulfur bacteria, Desulfovibrionaceae, and beta-Proteobacteria . Sequencing of the gene fragments confirmed that each primer pair was highly specific for the respective phylogenetic group . Applying the new primer sets, the bacterial diversity in the chemoclines of a eutrophic freshwater lake, a saline meromictic lake, and a laminated marine sediment was investigated . Compared to a conventional bacterial primer pair, a higher number of discrete DGGE bands was generated using our specific primer pairs . With one exception, all 15 bands tested yielded reliable 16S rRNA gene sequences . The highest diversity was found within the chemocline microbial community of the eutrophic freshwater lake . Sequence comparison revealed that the six sequences of Chromatiaceae and green sulfur bacteria detected in this habitat all represent distinct and previously unknown phylotypes . The lowest diversity of phylotypes was detected in the chemocline of the meromictic saline lake, which yielded only one sequence each of the Chromatiaceae, beta-2-Proteobacteria, and Desulfovibrionaceae, and no sequences of green sulfur bacteria . The newly developed primer sets are useful for the detection of previously unknown phylotypes, for the comparison of the microbial diversity between different natural habitats, and especially for the rapid monitoring of enrichments of unknown bacterial species. J Immunol, 1999 Aug 1, 163(3), 1570 - 6 Generation of nitric oxide by the inducible nitric oxide synthase protects gamma delta T cells from Mycobacterium tuberculosis-induced apoptosis; Sciorati C et al.; Gamma delta T cells are early recruited into mycobacterial lesions . Upon microbial Ag recognition, gamma delta cells secrete cytokines and chemokines and undergo apoptosis via CD95/CD95 ligand (CD95L) interaction, possibly influencing the outcome of infection and the characteristics of the disease . In this paper we show that activated phagocytes acquire, upon challenge with Mycobacterium tuberculosis, the ability to inhibit M . tuberculosis-induced gamma delta cell apoptosis . Apoptosis protection was due to NO because it correlated with NO synthase (NOS)-2 induction and activity in scavenger cells and was abrogated by NOS inhibitors . Furthermore, the NO donor S-nitrosoacetylpenicillamine mimicked the effect of enzyme induction . NO left unaffected the expression of CD95 and CD95L, suggesting interference with an event ensuing CD95/CD95L interaction . NO was found to interfere with the intracellular accumulation of ceramide and the activation of caspases, which were involved in gamma delta T cells apoptosis after M . tuberculosis recognition . We propose that NO generated by infected macrophages determines the life span and therefore the function of lymphocytes at the infection site, thus linking innate and adaptive immunity. Med Parazitol (Mosk), 1999 Jan-Mar, (1), 55 - 8 {The survival of the causative agent of plaque in the long-tailed suslik from a Tuva natural focus in wintertime}; Maevskii MP et al.; The survival of the causative agent of plague in the long-tailed souslik in the Tuva natural focus in winter was experimentally studied . They were made in a special bunker laboratory just in the focus . The experimental conditions were close to the hygrothermal parameters of a long-tailed souslik's burrow . Inoculation and placement of the animals and fleas into the bunker were accomplished in the September to early October . The rodents and ectoparasites were examined after their hibernation in the late April to early May of the following year . The duration of the experiment was 7.5 months . It has been found that the long-tailed souslik can be infected with the causative agent of plague before hibernation through transmission . There were no cases of plague microbial infection through the bite of fleas in sousliks following hibernation . Low infection rates of the fleas hibernating with their host were notified . The causative agent was found to survive in the mummified carcasses of sousliks for 7.5 months (the follow-up period). Plant Mol Biol, 1999 May, 40(2), 323 - 32 A novel E-type endo-beta-1,4-glucanase with a putative cellulose-binding domain is highly expressed in ripening strawberry fruits; Trainotti L et al.; Two full-length cDNA clones (faEG1 and faEG3, respectively) have been isolated by screening a cDNA library representing transcripts from red strawberry fruits . Southern blot analysis of genomic DNA suggests that the strawberry endo-beta-1,4-glucanases (EGases) are encoded by a multigene family . The cognate genes are predominantly expressed during the ripening process proper, although, in the case of faEG3, some expression has also been observed in large green fruits and, at low amounts, in young vegetative green tissues . In agreement with other ripening-related genes in strawberry, also the expression of faEG1 and faEG3 is down-regulated by treatment with an auxin analogue (1-naphthaleneacetic acid, NAA) . Differences in temporal expression of the two EGase genes in fruits are not accompanied by differences in spatial expression . The pattern of expression and the sequence characteristics of the two polypeptides suggest that the two strawberry EGases operate in a synergistic and coordinate manner . The protein encoded by faEG1 looks like one of the usual higher-plant EGases (average molecular mass of 54 kDa), while the protein encoded by faEG3 has a greater deduced molecular mass (about 68 kDa) due to the presence of an extra peptide of about 130 amino acids at the C-terminus . Such unusual peptide shows some features also found in microbial cellulases and contains a putative cellulose-binding domain . We propose that the faEG3-encoded EGase might especially hydrolyse the xyloglucans coating the cellulose microfibrils, thus rendering the cell wall more susceptible to the subsequent hydrolytic activity of the faEG1-encoded EGase. Croat Med J, 1999 Sep, 40(3), 392 - 7 Histological analysis and ancient DNA amplification of human bone remains found in caius iulius polybius house in pompeii; Cipollaro M et al.; Thirteen skeletons found in the Caius Iulius Polybius house, which has been the object of intensive study since its discovery in Pompeii 250 years ago, have provided an opportunity to study either bone diagenesis by histological investigation or ancient DNA by polymerase chain reaction analysis . DNA analysis was done by amplifying both X- and Y-chromosomes amelogenin loci and Y-specific alphoid repeat locus . The von Willebrand factor (vWF) microsatellite locus on chromosome 12 was also analyzed for personal identification in two individuals showing alleles with 10/11 and 12/12 TCTA repeats, respectively . Technical problems were the scarcity of DNA content from osteocytes, DNA molecule fragmentation, microbial contamination which change bone structure, contaminating human DNA which results from mishandling, and frequent presence of Taq DNA polymerase inhibiting molecules like polyphenols and heavy metals . The results suggest that the remains contain endogenous human DNA that can be amplified and analyzed . The amplifiability of DNA corresponds to the bone preservation and dynamics of the burial conditions subsequent to the 79 A.D . eruption. JAMA, 1999 Jul 14, 282(2), 175 - 81 The rational clinical examination . Does this adult patient have acute meningitis? Attia J, Hatala R, Cook DJ, Wong JG. CONTEXT: Early clinical recognition of meningitis is imperative to allow clinicians to efficiently complete further tests and initiate appropriate therapy . OBJECTIVE: To review the accuracy and precision of the clinical examination in the diagnosis of adult meningitis . DATA SOURCES: A comprehensive review of English- and French-language literature was conducted by searching MEDLINE for 1966 to July 1997, using a structured search strategy . Additional references were identified by reviewing reference lists of pertinent articles . STUDY SELECTION: The search yielded 139 potentially relevant studies, which were reviewed by the first author . Studies were included if they described the clinical examination in the diagnosis of objectively confirmed bacterial or viral meningitis . Studies were excluded if they enrolled predominantly children or immunocompromised adults or focused only on metastatic meningitis or meningitis of a single microbial origin . A total of 10 studies met the criteria and were included in the analysis . DATA EXTRACTION: Validity of the studies was assessed by a critical appraisal of several components of the study design . These components included an assessment of the reference standard used to diagnose meningitis (lumbar puncture or autopsy), the completeness of patient ascertainment, and whether the clinical examination was described in sufficient detail to be reproducible . DATA SYNTHESIS: Individual items of the clinical history have low accuracy for the diagnosis of meningitis in adults (pooled sensitivity for headache, 50% {95% confidence interval {CI}, 32%-68%}; for nausea/vomiting, 30% {95% CI, 22%-38%}) . On physical examination, the absence of fever, neck stiffness, and altered mental status effectively eliminates meningitis (sensitivity, 99%-100% for the presence of 1 of these findings) . Of the classic signs of meningeal irritation, only 1 study has assessed Kernig sign; no studies subsequent to the original report have evaluated Brudzinski sign . Among patients with fever and headache, jolt accentuation of headache is a useful adjunctive maneuver, with a sensitivity of 100%, specificity of 54%, positive likelihood ratio of 2.2, and negative likelihood ratio of 0 for the diagnosis of meningitis . CONCLUSIONS: Among adults with a clinical presentation that is low risk for meningitis, the clinical examination aids in excluding the diagnosis . However, given the seriousness of this infection, clinicians frequently need to proceed directly to lumbar puncture in high-risk patients . Many of the signs and symptoms of meningitis have been inadequately studied, and further prospective research is needed. Eur J Biochem, 1999 Jun, 262(3), 915 - 23 The inhibitory properties and primary structure of a novel serine proteinase inhibitor from the fruiting body of the basidiomycete, Lentinus edodes; Odani S et al.; A novel proteinase inhibitor, Lentinus proteinase inhibitor, has been purified from the fruiting bodies of the edible mushroom, Lentinus edodes, by buffer extraction and affinity chromatography on immobilized anhydrotrypsin . The protein simultaneously inhibits bovine beta-trypsin and alpha-chymotrypsin at independent sites, with apparent dissociation constants of 3.5 x 10(-10) M and 4 x 10(-8) M, respectively . The purified protein is eluted as two well-separated peaks on reversed-phase HPLC, one of which is inhibitory-active and the other inactive, and they are interconvertible under folding/unfolding conditions . Among the mammalian and microbial serine proteinases examined, including human enzymes of blood coagulation and fibrinolysis, activated factor XI was inhibited by the Lentinus proteinase inhibitor . Chemical modification studies suggest involvement of one or more arginine residues in the inhibition of trypsin . The complete primary structure composed of 142 amino acids with an acetylated N-terminus was determined by protein analysis . The theoretical molecular mass (15999.2) from the sequence is close to the experimental value of 15999.61 +/- 0.61 determined by mass spectrometry . Although there are no apparently homologous proteinase inhibitors in the protein database, there is a rather striking similarity to the propeptide segment of a microbial serine proteinase, as well as to the N-terminal region of the mature enzyme. Curr Opin Rheumatol, 1999 Jul, 11(4), 251 - 6 Psoriatic arthritis and the spectrum of syndromes related to the SAPHO (synovitis, acne, pustulosis, hyperostosis, and osteitis) syndrome; Winchester R; During the past year, the increasing use of nuclear magnetic resonance imaging techniques, with their ability to delineate cartilage and ligamentous structures and to identify edema, are providing a radical improvement in ascertainment of musculoskeletal abnormalities, although their significance remains incompletely delineated . A second theme has come from the study of spondyloarthropathies in different ethnic groups and societal environments, revealing that the Northern European and North American form of the disease, with its powerful association with HLA-B27, is little evident in the rest of the world's population and that different susceptibility genes and environmental factors operate in other regions and peoples . Related to this theme is the compelling evidence of the marked influence of HIV infection on the development of spondyloarthropathies in Africa . Two areas of immune recognition are discussed as examples of emerging fields that may provide useful paradigms for the experimental approach to mechanisms in psoriatic arthritis . One of these is the three-cell model of CD8 T-cell interaction, in which a dendritic cell presents a peptide from an immunogenic protein to both a CD4 and CD8 T-cell clone, providing a cognitive interaction that disrupts tolerance and results in the expansion of the cytotoxic T-cell clone . In this respect, the combination of an activated dendritic cell, together with enhanced availability of arthritogenic microbial antigens caused by microbial persistence, are interesting candidates to explore as the basis of the HIV-associated rheumatic diseases . The second area of immune recognition is the growing understanding of the outline of the solution to the problem of the association of a spondyloarthropathy with several J Ophthalmic Nurs Technol, 1999 Mar-Apr, 18(2), 50 - 9; quiz 74-5 Practical ophthalmic microbiology; Levine J et al.; Infection is commonly encountered in everyday ophthalmic practice . It is of great importance that all ophthalmic personnel are familiar with the basic etiologies of these infections, and the basic techniques which are used in diagnosis . When evaluating a patient with possible infection, it is often of great help to be familiar with the normal microbial flora of the human body . This normal flora has been detailed in the article . The assumption is that knowledge of a patient's flora can help guide decisions about the possible etiologies of an infection . When presented with an obvious ocular infection, health care personnel should review the "usual suspects"--that is, the common etiologies for each type of infection . The key to combating ocular infections lies in accurate treatment of the presumed infectious agent . Therefore, the techniques and steps used in the identification of the etiology of an infection should be known to all ophthalmic personnel . The proper sterile techniques of obtaining a specimen from a suspected corneal ulcer or conjunctivitis and the plating of the specimen on specific agar for identification are essential to everyday practice (Figure 8) . Patients who will undergo surgery, regardless of the type, are being placed at a special risk for infection . These post-operative infections can destroy the work of the most careful and exact surgical technique . It is an essential part of the procedure that the patient be protected as best they can from subsequent infection . The techniques to reduce the risk of post-operative infection have been detailed above . Proper preparation of a sterile field, sterile draping, and perioperative antibiotics can reduce the chance of a subsequent infection . It is important that these basic steps of care be properly provided to help insure successful surgical outcomes and excellence in health care. Am J Physiol, 1999 Jul, 277(1 Pt 1), G1 - 5 Mucosal immunity and inflammation . III . The mucosal antigen barrier: cross talk with mucosal cytokines; Perdue MH; We have known for many years that mucosal responses to antigens are regulated by immune cells and their molecular signals . More recently, it has become clear that epithelial cells also synthesize and secrete chemokines and cytokines . A sophisticated system of bidirectional cytokine signals is responsible for immune activation in the case of enteropathogens vs . immune suppression to food and commensal microbial antigens . A key factor in determining antigen handling is the route taken by antigens across the epithelial barrier . Cytokines and other mucosal messenger molecules play a critical role in the regulation of transepithelial antigen transport. Rapid Commun Mass Spectrom, 1999 Jul, 13(13), 1311 - 1314 Carbon stable isotopes reveal complex trophic interactions in lake plankton; Grey J et al.; The lower trophic linkages in lake plankton food webs are generally described as relatively simple, even accounting for the additional complexity of potential 'microbial looping' . Crustacean zooplankton are frequently amalgamated into one trophic functional group as grazers of autotrophic production . The carbon stable isotope ratios for separated zooplankton species, particulate organic matter (POM) and phytoplankton from a number of lakes in Finland and the UK were analysed . These revealed greater complexity in trophic interactions than would otherwise be observed if the zooplankton had been represented by a mixed sample . Grazing zooplankton were usually depleted in (13)C relative to the bulk POM on which they might feed, with (13)C deviating by up to 17 per thousand There were no consistent differences between (13)C values for copepods and cladocerans . Predatory cladocerans were generally enriched by greater than 1 per thousand compared to their putative prey . We suggest that care in separating the zooplankton species for stable isotope analysis may expose otherwise undetected sources of carbon and facilitate unravelling trophic links further up the food web . Rapid Commun Mass Spectrom, 1999, 13(13), 1291 - 4 Natural 13C abundance: a tool to trace the incorporation of dung-derived carbon into soil particle-size fractions; Amelung W et al.; During the decay of 13C enriched dung patches, the; delta 13C signal of surface soil (1-5 cm) increased with a temporary maximum after 42 d . To understand the underlying processes, we investigated the incorporation of dung-derived C into soil particle-size fractions . Dung, collected from beef steers fed on maize (delta 13C = -15.36/1000) or ryegrass (delta 13C = -25.67/1000), was applied in circular patches to a C3 pasture at North Wyke, UK . Triplicates were sampled from surface soil (1-5 cm) at 14, 28, 42, and 70 d after application, pooled, separated into fine (< 0.2 micron) and coarse clay (0.2-2 microns), silt plus fine sand (2-250 microns), and coarse sand (250-2000 microns), and analyzed for total C, N, and delta 13C . As particle-size diameter decreased, the C/N ratios decreased and delta 13C values increased at all plots due to increasing microbial alteration of soil organic matter . After dung application, ca . 60% of dung-derived C in soil was recovered in the 0.2-250 microns fractions during the whole experiment . The proportion of dung-derived C in the fine clay peaked 42 d after dung application, coinciding with the delta 13C maximum in the bulk soil and the maximum leaching rate measured in lysimeters at this time in another study at the same sites . The percentage of dung-derived C as particulate C in the coarse sand fraction increased until the end of the experiment . We conclude that incorporation of C into soil from decomposing dung patches involved both temporary sorption of leached dung C to < 0.2 micron fractions and continuous accumulation of particulate C (> 250 microns). Rapid Commun Mass Spectrom, 1999 Jul, 13(13), 1284 - 1290 Isotopic ((13)C) fractionation during plant residue decomposition and its implications for soil organic matter studies; Schweizer M et al.; Carbon isotopic fractionations in plant materials and those occurring during decomposition have direct implications in studies of short-and longer-term soil organic matter dynamics . Thus the products of decomposition, the evolved CO(2) and the newly formed soil organic matter, may vary in their (13)C signature from that of the original plant material . To evaluate the importance of such fractionation processes, the variations in (13)C signatures between and within plant parts of a tropical grass (Brachiaria humidicola) and tropical legume (Desmodium ovalifolium) were measured and the changes in (13)C content (signatures) during decomposition were monitored over a period of four months . As expected the grass materials were less depleted in (13)C (-11.4 to -11.9 per thousand) than those of the legume (-27.3 to -25.8 per thousand) . Root materials of the legume were less (1.5 per thousand) depleted in (13)C compared with the leaves . Plant lignin-C was strongly depleted in (13)C compared with the bulk material by up to 2.5 per thousand in the legume and up to 4.7 per thousand in the grass . Plant materials were subsequently incubated in a sand/nutrient-solution/microbial inoculum mixture . The respiration product CO(2) was trapped in NaOH and precipitated as CaCO(3), suitable for analysis using an automated C/N analyser coupled to an isotope ratio mass spectrometer . Significant depletion in (13)C of the evolved CO(2) was observed during the initial stages of decomposition probably as a result of microbial fractionation as it was not associated with the (13)C signatures of the measured more decomposable fractions (non-acid detergent fibre and cellulose) . While the cumulative CO(2)-(13)C signatures of legume materials became slightly enriched with ongoing decomposition, the CO(2)-C of the grass materials remained depleted in (13)C . Associated isotopic fractionation correction factors for source identification of CO(2-)C varied with time and suggested errors of 2-19% in the estimation of the plant-derived C at 119 days of incubation in a soil of an intermediate (-20.0 per thousand) (13)C signature . Analysis of the residual material after 119 days of incubation showed little or no change in the (13)C signature partly due to the incomplete decomposition at the time of harvesting . Glycobiology, 1999 Aug, 9(8), 747 - 55 Evolutionary considerations in relating oligosaccharide diversity to biological function; Gagneux P et al.; The oligosaccharide chains (glycans) attached to cell surface and extracellular proteins and lipids are known to mediate many important biological roles . However, for many glycans, there are still no evident functions that are of obvious benefit to the organism that synthesizes them . There is also no clear explanation for the extreme complexity and diversity of glycans that can be found on a given glycoconjugate or cell type . Based on the limited information available about the scope and distribution of this diversity among taxonomic groups, it is difficult to see clear trends or patterns consistent with different evolutionary lineages . It appears that closely related species may not necessarily share close similarities in their glycan diversity, and that more derived species may have simpler as well as more complex structures . Intraspecies diversity can also be quite extensive, often without obvious functional relevance . We suggest one general explanation for these observations, that glycan diversification in complex multicellular organisms is driven by evolutionary selection pressures of both endogenous and exogenous origin . We argue that exogenous selection pressures mediated by viral and microbial pathogens and parasites that recognize glycans have played a more prominent role, favoring intra- and interspecies diversity . This also makes it difficult to appreciate and elucidate the specific endogenous roles of the glycans within the organism that synthesizes them. Neurol Res, 1999 Jun, 21(4), 391 - 8 Effects of microbial invasion on cerebral hemodynamics and oxygenation monitored by near infrared spectroscopy in experimental Escherichia coli meningitis in the newborn piglet; Park WS et al.; This study was carried out to elucidate the pathophysiologic mechanism of cerebral hyperemia observed during the early phase of bacterial meningitis . We tested the hypothesis that microbial invasion through the blood-brain barrier is responsible for cerebral vasodilation and hyperemia in meningitis . Escherichia coli was given either intravenously (i.v.) or intracisternally (i.c.) to closely mimic the primary or secondary bacterial invasion occurring in meningitis and newborn piglets were grouped according to their invasion results (+ or -); 12 in the i.v . (+) group, 14 in the i.v . (-) group, 13 in the i.c . (+) group, 15 in the i.c . (-) group . The results were compared with eight animals in the control group . Near infrared spectroscopy (NIRS) was employed to monitor changes in total hemoglobin (HbT), oxygenated hemoglobin (HbO), deoxygenated hemoglobin (Hb), deduced hemoglobin (HbD), and oxidized cytochrome aa3 (Cyt aa3) . HbT, as an index of cerebral blood volume, increased progressively in both i.v . (+) and i.v . (-) groups and became significantly different from control and baseline values at 2 h . Hb significantly increased only in i.v . (+) group . HbD, as an index of cerebral blood flow, decreased significantly in i.v . (+), i.v.(-) and i.c . (-) groups and this change was mitigated in i.c . (+) group, HbO was reduced in i.c . (-) group and this decrease was attenuated in i.c . (+) group . Increased Cyt aa3 was observed in all experimental groups after bacterial inoculation . Changes in ICP, blood pressure, cerebral perfusion pressure, blood or CSF glucose or lactate, CSF TNF-alpha level, or CSF leukocytes number were not associated with changes in NIRS findings . These findings suggest that primary or secondary bacterial invasion across the blood-brain barrier is primarily responsible for cerebral vasodilation and hyperemia observed during the early phase of bacterial meningitis. Mol Immunol, 1999 Mar-Apr, 36(4-5), 249 - 60 Evasion of pathogens by avoiding recognition or eradication by complement, in part via molecular mimicry; Wurzner R; Most pathogens invading the human body are attacked by the host immune system directly following entry and usually also during most stages of the disease, especially when they are in contact with the blood . However, pathogens have developed an effective battery of specific strategies to overcome immune defense . This, far from being complete, review concentrates on evasion of pathogens by avoiding recognition or eradication by complement . The latter is achieved by removal of complement either by shedding it off the microbial surface, by consuming it away from the target membrane or by destroying it . Alternative procedures of avoiding eradication are the inhibition of complement activation or the employment of complement proteins via several highly sophisticated mechanisms, including the imitation of complement-like proteins (molecular mimicry). Jpn J Thorac Cardiovasc Surg, 1999 May, 47(5), 199 - 203 Does preoperative chemotherapy cause adverse effects on the perioperative course of patients undergoing esophagectomy for carcinoma? Baba M, Natsugoe S, Shimada M, Nakano S, Shirao K, Kusano C, Fukumoto T, Aikou T. The aim of this study was to clarify whether preoperative chemotherapy caused adverse effects on the perioperative course of patients undergoing esophagectomy . A total of 42 esophageal cancer patients were entered into a randomized trial and were analyzed . Twenty-one patients were assigned to immediate surgery (Surgery Group) . The other 21 received two 5-day courses of chemotherapy comprising cisplatin (70 mg/m2) on day 1, and fluorouracil (700 mg/m2) and leucovorin (20 mg/m2) on each of days 1 to 5 (chemotherapy group) . Hospital mortality comprised of one patient (2.3%) who had undergone an operation in the beginning of this series at 21 days after chemotherapy . Thereafter, the interval between the chemotherapy and operation was prolonged, with the average being 35 +/- 7 days . Preoperatively, both the lymphocyte counts and serum albumin levels were not increased in the chemotherapy group of patients even though their body weights increased . In the chemotherapy group, the operation time and the blood loss were increased and, on the 1st postoperative day, the development of systemic inflammatory response syndrome was high but the level of C-reactive protein was low . The incidence of positive microbial cultures of sputum and/or wound discharge within 8 postoperative days was higher in the chemotherapy group (42.9%) than in the surgery group (4.8%) . The host defense damage caused by chemotherapy may be prolonged and may show adverse effects in patients undergoing esophagectomy in the early postoperative period . Minimally, a 4-week interval between the completion of chemotherapy and operation is recommended for preventing surgical mortality related to the preoperative chemotherapy. J Biol Chem, 1999 Jul 16, 274(29), 20578 - 86 Broad spectrum thiopeptide recognition specificity of the Streptomyces lividans TipAL protein and its role in regulating gene expression; Chiu ML et al.; Microbial metabolites isolated in screening programs for their ability to activate transcription of the tipA promoter (ptipA) in Streptomyces lividans define a class of cyclic thiopeptide antibiotics having dehydroalanine side chains ("tails") . Here we show that such compounds of heterogeneous primary structure (representatives tested: thiostrepton, nosiheptide, berninamycin, promothiocin) are all recognized by TipAS and TipAL, two in-frame translation products of the tipA gene . The N-terminal helix-turn-helix DNA binding motif of TipAL is homologous to the MerR family of transcriptional activators, while the C terminus forms a novel ligand-binding domain . ptipA inducers formed irreversible complexes in vitro and in vivo (presumably covalent) with TipAS by reacting with the second of the two C-terminal cysteine residues . Promothiocin and thiostrepton derivatives in which the dehydroalanine side chains were removed lost the ability to modify TipAS . They were able to induce expression of ptipA as well as the tipA gene, although with reduced activity . Thus, TipA required the thiopeptide ring structure for recognition, while the tail served either as a dispensable part of the recognition domain and/or locked thiopeptides onto TipA proteins, thus leading to an irreversible transcriptional activation . Construction and analysis of a disruption mutant showed that tipA was autogenously regulated and conferred thiopeptide resistance . Thiostrepton induced the synthesis of other proteins, some of which did not require tipA. J Biol Chem, 1999 Jul 16, 274(29), 20165 - 70 The inhibition of capacitative calcium entry due to ATP depletion but not due to glucosamine is reversed by staurosporine; Vemuri S et al.; The capacitative Ca2+ entry pathway in J774 macrophages is rapidly inhibited by the amino sugar glucosamine . This pathway is also inhibited by treatments such as 2-deoxy-D-glucose (2dGlc) or glucose deprivation that inhibit glycolysis and lead to significant decreases in cellular ATP and other trinucleotides . We sought to determine whether glucosamine's effect on capacitative Ca2+ entry was also due to ATP depletion, as has been suggested recently for its link to insulin resistance . In contrast to brief treatments with 2dGlc, there was no significant decrease in ATP following exposure to glucosamine . In addition, the 2dGlc-mediated inhibition of capacitative Ca2+ influx was reversed by staurosporine, a microbial alkaloid that inhibits a broad range of protein kinases . Staurosporine was also able to reverse the inhibition of capacitative Ca2+ entry seen following other treatments that decreased cellular ATP levels, including cytochalasin B and iodoacetic acid . Other inhibitors of protein kinase C, including bisindolylmaleimide, K252a, H-7, and calphostin C, were unable to mimic this effect of staurosporine . However, the inhibition of capacitative Ca2+ influx in the presence of glucosamine was not reversed by staurosporine . These data indicate that the inhibitory action on capacitative Ca2+ entry of glucosamine is distinct from that caused by ATP depletion. Steroids, 1999 Mar, 64(3), 178 - 86 Recent advances in applied and mechanistic aspects of the enzymatic hydroxylation of steroids by whole-cell biocatalysts; Holland HL; Recent advances in microbial steroid hydroxylation are covered, including new biocatalysts and substrate groups and new methodologies such as the use of low-water systems, immobilised biocatalysts, genetically constructed biocatalysts and enzyme mimics . Mechanistic factors that control the regiochemistry and stereochemistry of steroid hydroxylation are also discussed. Amino Acids, 1999, 16(3-4), 191 - 213 Enzymatic resolution of amino acids via ester hydrolysis; Miyazawa T; The present review outlines recent examples of enzyme-based resolution procedures for amino acids via the hydrolysis of their esters . The resolutions have been achieved by using proteases (alpha-chymotrypsin, subtilisin and other microbial proteases, and sulfhydryl proteases of plant origin) and lipases . Relevant work utilizing yeast and other microbial cells is also included. J Mol Recognit, 1999 Jan-Feb, 12(1), 38 - 44 Receptins: a novel term for an expanding spectrum of natural and engineered microbial proteins with binding properties for mammalian proteins; Kronvall G et al.; A new term 'receptin', derived from recipere (lat.), is proposed to denote microbial binding proteins that interact with mammalian target proteins . An example of such a 'receptin' is staphyloccocal protein A which binds to the Fc part of many mammalian immunoglobulins . Several other types of 'receptins' are listed . This term may easily be distinguished from the similar term 'receptor', describing a binding site on a cell surface, mostly eukaryotic, where a secondary effect is induced inside the cell upon binding to a ligand . A receptin, however, does not necessarily have to induce a secondary event . Receptins include so called MSCRAMMs, adhesins, and also engineered receptins, affibodies, and engineered ligands . It denotes any protein of microbial origin, cell-bound or soluble, which can bind to a mammalian protein . It fulfills the need for an umbrella terminology for a large group of binding structures . In contrast, the term 'lectin' represents a group of proteins with affinity for carbohydrate structures . The new term 'receptin' includes a number of key microbial proteins involved in host-parasite interactions and in virulence . Some receptins are promising vaccine candidates . Biotechnol Bioeng, 1999 Jul 5, 64(1), 74 - 81 Thermodynamic analysis of growth of methanobacterium thermoautotrophicum Schill NA, Liu JS, Stockar Uv. Growth of Methanobacterium thermoautotrophicum, an anaerobic archaebacterium using methanogenesis as the catabolic pathway, is characterized by large heat production rates, up to 13 W g-1, and low biomass yields, in the order of 0.02 C-mol mol-1 H2 consumed . These values, indicating a possibly "inefficient" growth mechanism, warrant a thermodynamic analysis to obtain a better understanding of the growth process . The growth-associated heat production (DeltarHX0, min) and the growth-associated Gibbs energy dissipation per mol biomass formed (DeltarGXmin) were -3730 kJ C-mol-1 and -802 kJ C-mol-1, respectively . The Gibbs energy change found in this study is indeed unusually high as compared to aerobic methylotrophes, but not untypical for methanogens grown on CO2 . It explains the low biomass yield . Based on the information available on the energetic metabolism and on an ATP balance, the biomass yield can be predicted to be approximately in the range of the experimentally determined value . The fact that the exothermicity exceeds vastly even the Gibbs energy change can be explained by a dramatic entropy decrease of the catabolic reaction . Microbial growth characterized by entropy reduction and correspondingly by unusually large heat production may be called entropy-retarded growth . Biotechnol Bioeng, 1999 Jul 5, 64(1), 1 - 13 Efficiency of physical (light) or chemical (ABA, tetracycline, CuSO4 or 2-CBSU)-stimulus-dependent gus gene expression in tobacco cell suspensions; Boetti H et al.; In this study, the efficiency of inducible promoters to switch on gene expression in the presence of inducer or to switch it off in its absence was evaluated in tobacco cell suspensions transformed with the gus gene coding sequence . Either plant (pats1A, pSalT, pIn2-2) or microbial (pMre, pTet) inducible promoters were used to drive gus expression . The inducers were light, abscisic acid, 2-CBSU, CuSO4, tetracycline, respectively . For each construct (inducible promoter-gus coding sequence), the optimal induction conditions were determined (inducer concentration, induction time, and age of cells in culture cycle before induction) . The efficiency of the inducible promoter was then evaluated under optimal induction conditions . GUS-expression levels obtained under non-inducing and inducing conditions were systematically compared . Thirty or forty percent of the clones transformed with the pSalT-gus or pTet-gus construct, respectively, showed high induction rates (>1000) and GUS activities of the same order as those obtained with a constitutive system . However, basal GUS levels were always high for the pTet-gus cell lines . Seventy or eighty-five percent of the cell lines transformed with the pMre-gus or pln2-2-gus construct, respectively, had induction rates of 1.5 to 1000 . The pats1A-gus construct gave very low induction rates-55% of cell lines had induction rates less than 1.5 . Only the pSalt-gus construct gave both the highest induction rates and basal GUS-levels equivalent to the endogenous GUS background . J Periodontol, 1999 Jun, 70(6), 610 - 7 Opsonophagocytic effect of antibody against recombinant conserved 40-kDa outer membrane protein of Porphyromonas gingivalis; Saito S et al.; BACKGROUND: Porphyromonas gingivalis is associated with the initiation and progression of adult periodontitis . The outer membrane proteins of the bacteria are potentially important targets for interaction with host defense systems . A 40-kDa outer membrane protein (40-kDa OMP) is conserved among many strains of P . gingivalis . We have cloned the gene for 40-kDa OMP from P . gingivalis 381 and produced a recombinant protein . For the development of recombinant 40-kDa OMP as a component of a vaccine for passive immunization, the elucidation of the roles of the anti-recombinant 40-kDa OMP antibody in the host defense against P . gingivalis is essential . The objective of this study was to determine the opsonic capacity of the antibody for phagocytosis by neutrophils which play a key role in the immune response to microbial infections . METHODS: To test the opsonic activity of a rabbit polyclonal antibody against r40-kDa OMP (r40-kDa OMP Ab) on human neutrophils to phagocytize P . gingivalis, we constructed a reproducible in vitro model of P . gingivalis-neutrophil interaction using the human promyelocytic cell line HL-60 . RESULTS: We demonstrated that r40-kDa OMP Ab in the presence of human complement successfully opsonized {3H}-thymidine-labeled P . gingivalis as a target for phagocytosis by HL-60 cells differentiated with dimethyl sulfoxide . The phagocytized bacteria were then intracellularly killed and lysed, and the radioactive degradation debris egested into the culture medium . CONCLUSIONS: We conclude that antibody against r40-kDa OMP has opsonic activity on human neutrophil function for phagocytosis of P . gingivalis . Subgingival bacteria are coated in vivo with immunoglobulin and complement . When the antibody is specific for crevicular bacteria, immunological interactions can be expected in the crevice . Our observations suggest that the anti-recombinant 40-kDa OMP antibody in concert with the crevicular complement may prevent P . gingivalis colonization r40-kDa OMP may contribute to the development of a local immunotherapy when applied to the crevice of a patient with P . gingivalis-related periodontitis which relates to susceptibility for certain systemic diseases such as diabetes mellitus, cardiovascular disease, and preterm labor. Proc Natl Acad Sci U S A, 1999 Jul 6, 96(14), 7998 - 8002 The agricultural pathology of ant fungus gardens Currie CR, Mueller UG, Malloch D. Gardens of fungus-growing ants (Formicidae: Attini) traditionally have been thought to be free of microbial parasites, with the fungal mutualist maintained in nearly pure "monocultures." We conducted extensive isolations of "alien" (nonmutualistic) fungi from ant gardens of a phylogenetically representative collection of attine ants . Contrary to the long-standing assumption that gardens are maintained free of microbial pathogens and parasites, they are in fact host to specialized parasites that are only known from attine gardens and that are found in most attine nests . These specialized garden parasites, belonging to the microfungus genus Escovopsis (Ascomycota: anamorphic Hypocreales), are horizontally transmitted between colonies . Consistent with theory of virulence evolution under this mode of pathogen transmission, Escovopsis is highly virulent and has the potential for rapid devastation of ant gardens, leading to colony mortality . The specialized parasite Escovopsis is more prevalent in gardens of the more derived ant lineages than in gardens of the more "primitive" (basal) ant lineages . Because fungal cultivars of derived attine lineages are asexual clones of apparently ancient origin whereas cultivars of primitive ant lineages were domesticated relatively recently from free-living sexual stocks, the increased virulence of pathogens associated with ancient asexual cultivars suggests an evolutionary cost to cultivar clonality, perhaps resulting from slower evolutionary rates of cultivars in the coevolutionary race with their pathogens. J Microbiol Methods, 1999 Jul, 37(1), 17 - 22 Quantification of bacterial lead resistance via activity assays; Konopka A et al.; The level of microbial resistance to heavy metals is an important issue for the microbial ecology of heavy metal-contaminated habitats . However, assays based upon growth in nutrient media will overestimate the resistance level due to metal ion interactions with inorganic and organic components . The analysis of Pb-resistant bacteria isolated from soils containing up to 38 mmol total Pb x kg(-1) indicated that PYT80B medium which did not contain inorganic salts, contained low amounts of organic matter, and was buffered with a molecule that did not interact with metal ions (2-N-morpholinoethanesulfonic acid (MES)) provided the lowest estimates of lead resistance . However, better results were obtained by assaying metabolic activity (aerobic respiration) of resting cells suspended in 10 mM MES . By this criterion, 50% inhibition of Arthrobacter JS7 was found at 37 microM Pb(NO3)2 . The effects of Pb+2 concentrations upon respiration of resting cells and growth rate in PYT80B medium were similar . The activity assay also showed that metal resistance was induced to higher levels when Arthrobacter JS7 was grown in the presence of Pb. Am Fam Physician, 1999 Jun, 59(11), 3093 - 102, 3107-8 Kawasaki disease; Taubert KA et al.; Kawasaki disease is a leading cause of acquired heart disease among children in the United States and other developed countries . Most children who contract this illness are less than two years old, and 80 percent of affected children are younger than five years of age . A generalized vasculitis of unknown etiology, Kawasaki disease can cause coronary artery abnormalities, including coronary aneurysms . From 20 to 25 percent of untreated children develop coronary artery abnormalities, which may resolve or persist . These abnormalities are of particular concern because they can lead to thrombosis, evolve into segmental stenosis or, rarely, rupture . The principal cause of death from Kawasaki disease is myocardial infarction . The cause of the disease remains unknown, but epidemiologic investigations and the clinical presentation suggest a microbial agent . Diagnostic criteria, including fever and other principal features, have been established . In the acute phase of the disease, treatment with acetylsalicylic acid and intravenously administered immunoglobulin is directed at reducing inflammation of the coronary arteries and myocardium . Early recognition and treatment of Kawasaki disease can reduce the development of potentially life-threatening coronary artery abnormalities. Trends Microbiol, 1999 Jul, 7(7), 275 - 81 The cellulosome concept as an efficient microbial strategy for the degradation of insoluble polysaccharides; Shoham Y et al.; The cellulosome is an extracellular supramolecular machine that can efficiently degrade crystalline cellulosic substrates and associated plant cell wall polysaccharides . The cellulosome arrangement can also promote adhesion to the insoluble substrate, thus providing individual microbial cells with a direct competitive advantage in the utilization of the soluble hydrolysis products. J S C Med Assoc, 1999 Jun, 95(6), 227 - 30 The modifiable factors contributing to leading causes of death in South Carolina; Pilibosian ED et al.; In 1996, there were 34,035 deaths in South Carolina . Almost 70 percent of these deaths were due to chronic diseases . There are known ways to prevent chronic diseases from developing or at least delay their developmental process, thereby lengthening years of life . The purpose of this paper is to report modifiable risk factors for mortality related to leading causes of death . The top ten causes of death in South Carolina were obtained from the South Carolina Department of Health and Environmental Control . Estimates of the number of deaths due to certain modifiable risk factors were made using results of a study published by McGinnis and Foege . The percentage of deaths due to each cause was adapted to South Carolina death certificate data from the national estimates . Results indicate that small modifications in individual lifestyles could prevent or delay nearly 50 percent of deaths in South Carolina annually . Tobacco use, diet and physical activity, and misuse of alcohol contribute to the largest number of deaths . Other modifiable behaviors contributing to the 50 percent mortality are microbial agents, toxic agents, firearms, sexual behavior, motor vehicle accidents, and illicit use of drugs . The implication in these findings is that these risk factors for mortality are mainly modifiable . There are many causes of death that may be delayed due to these modifiable risk factors . By looking at preventable causes of death, rather than focusing on traditional causes of death, it becomes clear that prevention strategies are critically important. Biophys J, 1999 Jul, 77(1), 493 - 504 Molecular dynamics of microbial lipases as determined from their intrinsic tryptophan fluorescence; Graupner M et al.; We have studied the intrinsic tryptophan fluorescence of the lipases from Chromobacterium viscosum (CVL), Pseudomonas species (PSL), and Rhizopus oryzae (ROL) in aqueous buffer, zwitterionic detergent micelles, and isopropanol-water mixtures . It was the purpose of this study to obtain information about biophysical properties of the respective enzymes under conditions that modulate enzyme activities and stereoselectivities to a significant extent . According to their decay-associated emission spectra, CVL tryptophans are located in the hydrophobic interior of the protein . In contrast, the PSL and ROL tryptophans are probably confined to the core and the surface of the lipase . From the tryptophan lifetime distributions it can be concluded that the conformation of CVL is not much affected by detergent or organic solvent (isopropanol) . Accordingly, CVL is enzymatically active in these systems and most active in the presence of isopropanol . In contrast, ROL and PSL show high conformational mobility, depending on the solvent, because their lifetime distributions are very different in the presence and absence of detergent or isopropanol . Time-resolved anisotropy studies provided evidence that the lipases exhibit very high internal molecular flexibility . This peculiar feature of lipases is perhaps the key to the great differences in activity and stereoselectivity observed in different reaction media . Furthermore, information about self-association of the lipases in different solvents could be obtained . PSL, but not CVL and ROL, forms aggregates in water . Lipase aggregation can be reversed by the addition of detergent or isopropanol, which competes for the hydrophobic surface domains of this protein . This dissociation could efficiently contribute to the increase in lipase activity in the presence of a detergent or isopropanol. Appl Environ Microbiol, 1999 Jul, 65(7), 3164 - 74 Seasonal dynamics of bacterioplankton community structure in a eutrophic lake as determined by 5S rRNA analysis; Hofle MG et al.; Community structure of bacterioplankton was studied during the major growth season for phytoplankton (April to October) in the epilimnion of a temperate eutrophic lake (Lake Plusssee, northern Germany) by using comparative 5S rRNA analysis . Estimates of the relative abundances of single taxonomic groups were made on the basis of the amounts of single 5S rRNA bands obtained after high-resolution electrophoresis of RNA directly from the bacterioplankton . Full-sequence analysis of single environmental 5S rRNAs enabled the identification of single taxonomic groups of bacteria . Comparison of partial 5S rRNA sequences allowed the detection of changes of single taxa over time . Overall, the whole bacterioplankton community showed two to eight abundant (>4% of the total 5S rRNA) taxa . A distinctive seasonal succession was observed in the taxonomic structure of this pelagic community . A rather-stable community structure, with seven to eight different taxonomic units, was observed beginning in April during the spring phytoplankton bloom . A strong reduction in this diversity occurred at the beginning of the clear-water phase (early May), when only two to four abundant taxa were observed, with one taxon dominating (up to 72% of the total 5S rRNA) . The community structure during summer stagnation (June and July) was characterized by frequent changes of different dominating taxa . During late summer, a dinoflagellate bloom (Ceratium hirudinella) occurred, with Comamonas acidovorans (beta-subclass of the class Proteobacteria) becoming the dominant bacterial species (average abundance of 43% of the total 5S rRNA) . Finally, the seasonal dynamics of the community structure of bacterioplankton were compared with the abundances of other major groups of the aquatic food web, such as phyto- and zooplankton, revealing that strong grazing pressure by zooplankton can reduce microbial diversity substantially in pelagic environments. Appl Environ Microbiol, 1999 Jul, 65(7), 3064 - 70 Composition of toluene-degrading microbial communities from soil at different concentrations of toluene; Hubert C et al.; Toluene-degrading bacteria were isolated from hydrocarbon-contaminated soil by incubating liquid enrichment cultures and agar plate cultures in desiccators in which the vapor pressure of toluene was controlled by dilution with vacuum pump oil . Incubation in desiccators equilibrated with either 100, 10, or 1% (wt/wt) toluene in vacuum pump oil and testing for genomic cross-hybridization resulted in four genomically distinct strains (standards) capable of growth on toluene (strains Cstd1, Cstd2, Cstd5, and Cstd7) . The optimal toluene concentrations for growth of these standards on plating media differed considerably . Cstd1 grew best in an atmosphere equilibrated with 0.1% (wt/wt) toluene, but Cstd5 failed to grow in this atmosphere . Conversely, Cstd5 grew well in the presence of 10% (wt/wt) toluene, which inhibited growth of Cstd1 . 16S ribosomal DNA sequencing and cross-hybridization analysis indicated that both Cstd1 and Cstd5 are members of the genus Pseudomonas . An analysis of the microbial communities in soil samples that were incubated with 10% (wt/wt) toluene with reverse sample genome probing indicated that Pseudomonas strain Cstd5 was the dominant community member . However, incubation of soil samples with 0.1% (wt/wt) toluene resulted in a community that was dominated by Pseudomonas strain Q7, a toluene degrader that has been described previously (Y . Shen, L . G . Stehmeier, and G . Voordouw, Appl . Environ . Microbiol . 64:637-645, 1998) . Q7 was not able to grow by itself in an atmosphere equilibrated with 0.1% (wt/wt) toluene but grew efficiently in coculture with Cstd1, suggesting that toluene or metabolic derivatives of toluene were transferred from Cstd1 to Q7. Appl Environ Microbiol, 1999 Jul, 65(7), 2994 - 3000 Comparative diversity of ammonia oxidizer 16S rRNA gene sequences in native, tilled, and successional soils; Bruns MA et al.; Autotrophic ammonia oxidizer (AAO) populations in soils from native, tilled, and successional treatments at the Kellogg Biological Station Long-Term Ecological Research site in southwestern Michigan were compared to assess effects of disturbance on these bacteria . N fertilization effects on AAO populations were also evaluated with soils from fertilized microplots within the successional treatments . Population structures were characterized by PCR amplification of microbial community DNA with group-specific 16S rRNA gene (rDNA) primers, cloning of PCR products and clone hybridizations with group-specific probes, phylogenetic analysis of partial 16S rDNA sequences, and denaturing gradient gel electrophoresis (DGGE) analysis . Population sizes were estimated by using most-probable-number (MPN) media containing varied concentrations of ammonium sulfate . Tilled soils contained higher numbers than did native soils of culturable AAOs that were less sensitive to different ammonium concentrations in MPN media . Compared to sequences from native soils, partial 16S rDNA sequences from tilled soils were less diverse and grouped exclusively within Nitrosospira cluster 3 . Native soils yielded sequences representing three different AAO clusters . Probes for Nitrosospira cluster 3 hybridized with DGGE blots from tilled and fertilized successional soils but not with blots from native or unfertilized successional soils . Hybridization results thus suggested a positive association between the Nitrosospira cluster 3 subgroup and soils amended with inorganic N . DGGE patterns for soils sampled from replicated plots of each treatment were nearly identical for tilled and native soils in both sampling years, indicating spatial and temporal reproducibility based on treatment. Biochemistry, 1999 Jun 15, 38(24), 7678 - 88 Escherichia coli methionine aminopeptidase: implications of crystallographic analyses of the native, mutant, and inhibited enzymes for the mechanism of catalysis; Lowther WT et al.; By improving the expression and purification of Escherichia coli methionine aminopeptidase (eMetAP) and using slightly different crystallization conditions, the resolution of the parent structure was extended from 2.4 to 1.9 A resolution . This has permitted visualization of the coordination geometry and solvent structure of the active-site dinuclear metal center . One solvent molecule (likely a mu-hydroxide) bridges the trigonal bipyramidal (Co1) and octahedral (Co2) cobalt ions . A second solvent (possibly a hydroxide ion) is bound terminally to Co2 . A monovalent cation binding site was also identified about 13 A away from the metal center at an interface between the two subdomains of the protein . The first structure of a substrate-like inhibitor, (3R)-amino-(2S)-hydroxyheptanoyl-L-Ala-L-Leu-L-Val-L-Phe-OMe, bound to a methionine aminopeptidase, has also been determined . This inhibitor coordinates the metal center through four interactions as follows: (i) ligation of the N-terminal (3R)-nitrogen to Co2, (ii, iii) bridging coordination of the (2S)-hydroxyl group, and (iv) terminal ligation to Co1 by the keto oxygen of the pseudo-peptide linkage . Inhibitor binding occurs with the displacement of two solvent ligands and the expansion of the coordination sphere of Co1 . In addition to the tetradentate, bis-chelate metal coordination, the substrate analogue forms hydrogen bonds with His79 and His178, two conserved residues within the active site of all MetAPs . To evaluate their importance in catalysis His79 and His178 were replaced with alanine . Both substitutions, but especially that of His79, reduce activity . The structure of the His79Ala apoenzyme and the comparison of its electronic absorption spectra with other variants suggest that the loss in activity is not due to a conformational change or a defective metal center . Two different reaction mechanisms are proposed and are compared to those of related enzymes . These results also suggest that inhibitors analogous to that reported here may be useful in preventing angiogenesis in cancer and in the treatment of microbial and fungal infections. Microb Ecol, 1999 Jul, 38(1), 50 - 57 Decomposition and CO2 Evolution from Standing Litter of the Emergent Macrophyte Erianthus giganteus; Kuehn KA et al.; > Abstract Decomposition of standing litter of the emergent macrophyte Erianthus giganteus (plumegrass) was quantified in a small freshwater wetland in Alabama, USA . Living green shoots of E . giganteus were tagged and periodically retrieved for determination of leaf and culm mass loss, litter-associated fungal biomass (ergosterol), and nitrogen and phosphorus concentrations . Laboratory studies were also conducted to examine the effects of plant litter moisture content and temperature on rates of CO2 evolution from plant litter . Culm and leaf material lost 25 and 32% AFDM, respectively, during plant senescence and early litter decay . Fungal biomass, as determined by ergosterol concentrations, increased significantly in both leaf and culm litter during decomposition, with maximum biomass accounting for 3.7 and 6.7% of the total detrital weight in culm and leaf litter, respectively . Spatial differences in fungal biomass were observed along the culm axis, with upper regions of the culm accumulating significantly greater amounts of fungal mass than basal regions (p < 0.01, ANOVA) . Rates of CO2 evolution from both leaf and culm litter increased rapidly after wetting (0 to 76 microg CO2-C g-1 AFDM h-1 within 5 min) . In addition, rates of CO2 evolution from water saturated culms increased exponentially as the temperature was increased from 10 to 30 degrees C . These results provide evidence that considerable microbial colonization and mineralization of standing emergent macrophyte litter can occur before collapse of senescent shoot material to the water and sediment surface.http://link.springer-ny.com/link/service/journals/00248/bibs/38n1p50.html Curr Opin Microbiol, 1999 Jun, 2(3), 257 - 64 Biodesulfurization; McFarland BL; Microbial sulfur-specific transformations have been identified that selectively desulfurize organic sulfur compounds in fossil fuels . Recent discoveries related to biodesulfurization mechanisms may lead to commercial applications of biodesulfurization through engineering recombinant strains for over-expression of biodesulfurization genes, removal of end product repression, and/or by combining relevant industrial and environmental traits with improvements in bioprocess design. Curr Opin Microbiol, 1999 Jun, 2(3), 323 - 7 Terminal restriction fragment length polymorphism (T-RFLP): an emerging method for characterizing diversity among homologous populations of amplification products; Marsh TL; Terminal restriction fragment length polymorphism is a recent molecular approach that can assess subtle genetic differences between strains as well as provide insight into the structure and function of microbial communities . The technique has both high sensitivity and throughput making it ideal for comparative analyses. Curr Opin Microbiol, 1999 Jun, 2(3), 306 - 11 The role of short sequence repeats in epidemiologic typing; van Belkum A; Short sequence repeats (SSRs), also known as variable number of tandem repeats or micro-satellites, are inherently unstable entities that undergo frequent variation in the number of repeated units through slipped strand mispairing during DNA synthesis . In humans, unit number variability in SSRs has been associated with the occurrence of specific genetic diseases, whereas in micro-organisms SSRs have been elegantly linked to modulation of gene expression . Knowledge of the functional constraints imposed upon the SSRs sheds light on their potential use as molecular clocks for monitoring microbial genome evolution . Although microbial SSR genotypes have been used with increasing frequency for studying the epidemiology and evolution of microbial strains and isolates, such approaches should be used with caution. Gastroenterol Nurs, 1999 Mar-Apr, 22(2), 63 - 8 Microbial bioburden in endoscope reprocessing and an in-use evaluation of the high-level disinfection capabilities of Cidex PA; Vesley D et al.; Most endoscopy clinics use 2% glutaraldehyde as a high-level disinfectant for reprocessing flexible endoscopes . However, even with contact times greater than 30 minutes, survival of organisms has been documented . We compared the high-level disinfection capabilities of glutaraldehyde (45-minute immersion) with a new peracetic acid germicide (20- to 25-minute immersion) . Channels, valve housings, and outer sheaths were sampled to quantify bioburden levels after a patient procedure, after manual cleaning, and after disinfection . Total mean bioburden after clinical use was greater than 6 log10 colony-forming units (CFU) . Manual cleaning reduced the bioburden by means of 4.7 log10 CFU (gastroscopes) and 6.2 log10 CFU (colonoscopes) . High-level disinfection with the new product was achieved in five of six (product stressed by EPA Reuse Test) and 7 of 10 (product stressed by dilution and organic load) successfully disinfected endoscopes, whereas glutaraldehyde achieved it in 4 of 10 (product stressed by dilution and organic load) . We conclude that the new peracetic acid product (20- and 25-minute contact time) is at least as effective as glutaraldehyde (45-minute contact time) for reducing the bioburden of vegetative aerobic organisms in endoscopes. Biomaterials, 1999 Jun, 20(12), 1143 - 50 Mechanical loading of bovine pericardium accelerates enzymatic degradation; Ellsmere JC et al.; Bioprosthetic heart valves fail as the result of two simultaneous processes: structural deterioration and calcification . Leaflet deterioration and perforation have been correlated with regions of highest stress in the tissue . The failures have long been assumed to be due to simple mechanical fatigue of the collagen fibre architecture; however, we have hypothesized that local stresses-and particularly dynamic stresses-accelerate local proteolysis, leading to tissue failure . This study addresses that hypothesis . Using a novel, custom-built microtensile culture system, strips of bovine pericardium were subjected to static and dynamic loads while being exposed to solutions of microbial collagenase or trypsin (a non-specific proteolytic enzyme) . The time to extend to 30% strain (defined here as time to failure) was recorded . After failure, the percentage of collagen solubilized was calculated based on the amount of hydroxyproline present in solution . All data were analyzed by analysis of variance (ANOVA) . In collagenase, exposure to static load significantly decreased the time to failure (P < 0.002) due to increased mean rate of collagen solubilization . Importantly, specimens exposed to collagenase and dynamic load failed faster than those exposed to collagenase under the same average static load (P = 0.02) . In trypsin, by contrast, static load never led to failure and produced only minimal degradation . Under dynamic load, however, specimens exposed to collagenase, trypsin, and even Tris/CaCl2 buffer solution, all failed . Only samples exposed to Hanks' physiological solution did not fail . Failure of the specimens exposed to trypsin and Tris/CaCl2 suggests that the non-collagenous components and the calcium-dependent proteolytic enzymes present in pericardial tissue may play roles in the pathogenesis of bioprosthetic heart valve degeneration. Biopolymers, 1999 Aug, 50(2), 193 - 200 Enzymatic ligation for synthesis of single-chain analogue of monellin by transglutaminase; Ota M et al.; Monellin, a sweet protein, consists of two noncovalently associated polypeptide chains: an A chain of 44 amino acid residues and a B chain of 50 residues . Microbial transglutaminase (MTGase) was used for ligation of the monellin subunits without any protecting groups, and without activation of the C alpha-carboxyl group at the C-terminus . Since a peptide fragment LLQG is a good substrate for MTGase to form an amide bond between the gamma-amide group of the Gln residue and the epsilon-amino group of Lys, a monellin B chain analogue in which LLQG was elongated at the C-terminus (B-LLQG) was synthesized by solid-phase synthesis . The monellin A chain analogue in which KGK was elongated at the N-terminus (KGK-A) was synthesized by the same method as that of the B chain analogue . The KGK-A chain and the B-LLQG chain were coupled by MTGase to give single-chain analogue of monellin . The single-chain analogue of monellin was characterized by analytical reverse phase high performance liquid chromatography, electrospray ionization, and amino acid analyses . All analyses gave satisfactory results . The single-chain analogue of monellin was more heat stable than natural monellin. J Rheumatol, 1999 Jun, 26(6), 1306 - 11 Presence of antibodies against Helicobacter pylori and its heat-shock protein 60 in the serum of patients with Sjögren's syndrome; Aragona P et al.; OBJECTIVE: Helicobacter pylori infection elicits a local and systemic immune response against bacterial antigens, including a heat-shock protein of 60 kDa (HSP60) . The homology between microbial and human HSP suggests that the immune response to bacterial HSP may play a role in the pathogenesis of autoimmune disorders . Since gastric involvement and H . pylori have been reported in Sjogren's syndrome (SS), we investigated the prevalence of antibodies against H . pylori and its specific HSP60 in sera from patients with SS . METHODS: Four groups of patients were studied . Group 1, 34 patients with primary SS (pSS); Group 2.19 patients with secondary SS; Group 3, 22 patients with various autoimmune diseases and Group 4, 43 healthy controls . Serum IgG levels against HSP60 were determined by an ELISA using recombinant full length HSP60 expressed in Escherichia coli, as the antigen . To confirm the H . pylori infection, a commercial ELISA was used . RESULTS: Out of 34 patients in Group 1, 27 (79.4%) and 30 (88.2%) had antibodies against H . pylori and its HSP60, respectively . The prevalence was significantly higher than that found in Group 3 (18.2%, p < 0.0001 and 27.3%, p < 0.0001) and in Group 4 (48.8%, p < 0.005 and 37.2%, p < 0.0001) but not than that of Group 2 (48.8% and 37.2%) . If the prevalence of patients either positive or negative for both antibodies was considered, a statistically significant difference was found between Group I and respectively Groups 3 and 4 . CONCLUSION: The hypothetical role of HSP60 in the development of the immune response both in pSS and secondary SS seems strictly linked to the prevalence rate of H . pylori infection. Am J Dermatopathol, 1999 Jun, 21(3), 234 - 40 A clinical and histologic study of 37 cases of immunoglobulin A-associated vasculitis; Magro CM et al.; Immunoglobulin (Ig) A-associated vasculitis is commonly equated with the multiorgan systemic vasculitic syndrome Henoch-Schonlein purpura (HSP), which occurs predominantly in the pediatric age group . By natural language search of the databases of two outpatient dermatopathology practices, the authors selected for review 37 cases of IgA-associated vasculitis, 23 of which were associated with antecedent infection, most commonly of the upper respiratory tract . Criteria for a diagnosis of HSP were met in 15 cases, 13 of which were in the setting of prior infection . Lower extremity skin involvement was ubiquitous . A more widespread form of vasculitis was also seen, particularly in the setting of previous infection . Several of the patients with previous infection had underlying medical illnesses including rheumatoid arthritis, atopy, renal failure, lupus erythematosus, insulin dependent diabetes mellitus, autoimmune thyroid disease, and Wegener's granulomatosis . In those patients lacking an apparent microbial trigger, Sjogren's disease with anti-Ro antibodies and hypergammaglobulinemia, lupus erythematosus, inflammatory bowel disease, IgA paraproteinemia, bronchogenic and prostatic carcinoma, cryoglobulinemia, and lymphoma were uncovered . Regardless of whether an infectious stimulus was implicated, certain cofactors with the potential to enhance vascular injury were uncovered; these included anti-Ro antibodies, antineutrophil cytoplasmic antibody, diabetic microangiopathy, and a hyperviscosity state . In the infective group, a pustular vasculitis, defined as a neutrophilic vascular reaction in concert with epithelial pustulation, was seen in 81% of cases versus 33% in the noninfectious group (p = 0.02) . The prototypic histomorphology in the noninfective group was one of a mild cell poor leukocytoclastic vasculitis; Vasculitis was of greater severity in patients with antecedent infection (p = 0.026) . An infectious trigger, typically of mucosal origin, can frequently be identified in patients with cutaneous IgA-associated vasculitis, especially those with the symptom complex of HSP . The light microscopy appears to distinguish patients who have an infectious trigger from those who do not . IgA-associated vasculitis may be a clue to the presence of certain underlying disorders where there is immune dysregulation or enhanced susceptibility to immune complex entrapment. J Microbiol Methods, 1999 Jun, 36(3), 203 - 13 Comparison of two kinds of Biolog microplates (GN and ECO) in their ability to distinguish among aquatic microbial communities; Choi KH et al.; We compared the abilities of Biolog's GN and ECO plates to distinguish among aerobic and heterotrophic bacterial communities in samples from six aquatic environments . The Biolog system is based on interpreting patterns of sole-carbon substrate utilization indicated by color development in a 96-well microtiter plate . Whether of fresh or saltwater origin, bacterial communities utilized > 95% of substrates in both types of plates . Samples from any one environment exhibited similar time courses of average well color development (AWCD) in both GN and ECO plates . Principal component analysis was performed on data sets resulting from combinations of algorithms (AWCD and curve-integration methods) and levels of color development (end-point and set-point approaches) . In all cases, the two types of plates demonstrated an equal capacity to discriminate among the heterotrophic expressions of the six microbial communities . Substantial deviation from an anticipated 1:1 correspondence occurred when color development of 25 substrates common to both types of plates was compared . The discrepancies likely are related to the different formulations of low-nutrient media in GN and ECO plates. J Neuroimmunol, 1999 Jan 1, 93(1-2), 149 - 55 Synergistic interaction of catecholamine hormones and Mycobacterium avium results in the induction of interleukin-10 mRNA expression by murine peritoneal macrophages; Chen L et al.; The results of this investigation provides evidence that catecholamine hormones interact with macrophages that are infected with Mycobacterium avium resulting in the induction of IL-10 mRNA and protein . The effect of catecholamine hormones was prevented by treating the cells with the beta-adrenergic receptor antagonist propranolol but not by alpha-adrenergic antagonist phentolamine . The effect of catecholamine stimulation was mimicked by the addition of beta-2 adrenergic agonists and by the addition of cAMP to the infected macrophage cultures . These observations suggest that sympathetic nervous system activation together with microbial infection results in a synergistic interaction that could result in the control of inflammatory processes. FEMS Immunol Med Microbiol, 1999 Jun, 24(2), 193 - 9 Molecular methods for typing of Helicobacter pylori and their applications; Colding H et al.; Microbial typing is a useful tool in clinical epidemiology for defining the source and route of infection, for studying the persistence and reinfection rates, clonal selection in the host and bacterial evolution . Phenotypic methods such as biotyping, serotyping and hemagglutinin typing have little discriminatory power compared to genotypic methods concerning the typing of Helicobacter pylori . Therefore great efforts have been made to establish useful molecular typing methods . In this context, the most frequently used genotypic methods are described based on our own experience and the literature: (1) restriction endonuclease analysis, (2) endonuclease analysis using pulsed-field gel electrophoresis, (3) ribotyping, (4) polymerase chain reaction (using either random primers or repetitive DNA sequence primers), and (5) polymerase chain reaction-restriction fragment length polymorphism analysis of e.g . the urease genes . Furthermore, reproducibility, discriminatory power, ease of performance and interpretation, cost and toxic procedures of each method are assessed . To date no direct comparison of all the molecular typing methods described has been performed in the same study with the same H . pylori strains . However, PCR analysis of the urease gene directly on suspensions of H . pylori or gastric biopsy material seems to be useful for routine use and applicable in specific epidemiological situations. Bioessays, 1999 May, 21(5), 377 - 81 The genome of Rickettsia prowazekii and some thoughts on the origin of mitochondria and hydrogenosomes; Muller M et al.; The sequence of an alpha-proteobacterial genome, that of Rickettsia prowazekii, is a substantial advance in microbial and evolutionary biology . The genome of this obligately aerobic intracellular parasite is small and is apparently still undergoing reduction, reflecting gene losses attributable to its intracellular parasitic lifestyle . Evolutionary analyses of proteins encoded in the genome contain the strongest phylogenetic evidence to date for the view that mitochondria descend from alpha-proteobacteria . Although both Rickettsia and mitochondrial genomes are highly reduced, it appears that genome reduction in these lineages has occurred independently . Rickettsia's genome encodes an ATP-generating machinery that is strikingly similar to that of aerobic mitochondria . But it does not encode homologues for the ATP-producing pathways of anaerobic mitochondria or hydrogenosomes, leaving an important issue regarding the origin and nature of the ancestral mitochondrial symbiont unresolved. J Anim Sci, 1999 Jun, 77(6), 1587 - 96 Influence of grain source on ruminal characteristics and rate, site, and extent of digestion in beef steers; Philippeau C et al.; Six cannulated Salers steers (305 +/- 17 kg initial BW) were used in a double 3 x 3 Latin square design to compare the effects of the nature of the cereal (wheat vs corn) and the corn genotype (dent vs flint) on rate, site, and extent of digestion of high-concentrate diets . The cereals were coarsely cracked, and the diets were balanced to have the same percentage of starch (47.7 +/- 2.3%) and CP (14.6 +/- .7%) . Differences in ruminal starch digestion were observed between wheat- and corn-based diets (86.6 vs 47.8%; P < .001) and between corn genotypes (60.8 vs 34.8% for dent and flint corns; P < .001) . For flint corn, more than half the starch was digested in the hindgut . Total tract digestion of starch was greater (P < .001) by steers fed wheat than by those fed corn and did not differ (P > .1) between the two corn genotypes . Ruminal mean pH (P < .01) was lower and total VFA concentration (P < .1) was higher for wheat- than for corn-based diets . Ruminal acetate:propionate tended to increase with the decrease in the amount of starch degraded in the rumen, but differences were not significant (P > .1) . When wheat replaced corn, nonammonia, nonmicrobial N duodenal flow decreased (P < .01), and microbial duodenal flow increased (P < .05), so there were no differences in the duodenal flow of nonammonia N duodenal flow (P > .1) . The lower nonammonia N duodenal flow for the dent corn- than for the flint corn-based diet (P < .05) was related to a lower passage of nonammonia, nonmicrobial N into the duodenum . Efficiency of microbial protein synthesis was inversely correlated with the amount of starch degraded in the rumen . Nature of the cereal, wheat vs corn, and genotype of the corn, dent vs flint, alter the site and extent of starch digestion. J Anim Sci, 1999 Jun, 77(6), 1565 - 71 In situ neutral detergent insoluble nitrogen as a method for measuring forage protein degradability; Mass RA et al.; A method of estimating the undegraded intake protein (UIP) concentration of forages was developed and validated with a series of in situ experiments . The hypothesis was that UIP calculated from in situ neutral detergent insoluble N (NDIN) is equal to total in situ N minus the microbial N that is estimated from purines (MN) . The in situ disappearance rates of total in situ N (TN), MN, and NDIN were measured for six hay samples and two range masticate samples . Hypothetical rates of passage (2 or 5%/h) were used to calculate UIP (% of DM) for each N pool . Estimates of UIP from TN were higher (P = .0001) than those from either MN or NDIN, and MN estimates of UIP were similar (P = .48) to NDIN estimates . A low-N fiber source (solka floc) was incubated in situ for 8 h . Analysis of the residue detected purines before, but not after, neutral detergent extraction . Several in situ incubation (i.e., Dacron bag size and number of Dacron bags in a mesh bag) and neutral detergent extraction conditions were tested . None of the factors tested affected in situ NDIN disappearance (P > .05) . The hypothesis that NDIN is completely digestible in the rumen was tested . Estimates of the extent of NDIN digestion were made using 96-h in situ incubations, and UIP was recalculated for the test samples . Mean in situ UIP concentration decreased upon recalculation (P = .05) . In situ NDIN provides estimates of forage UIP that are equal to estimates from MN . Forage UIP estimates are less when extent of N degradation is estimated and included in the calculation. Ann Acad Med Singapore, 1999 Jan, 28(1), 120 - 32 Immunological consequences of trauma and shock; Catania RA et al.; The immune system is a powerful, complex entity composed of numerous cell types and regulated by autocrine, paracrine, and hormonal mechanisms . Trauma and haemorrhagic shock induce numerous changes within this system which are ultimately deleterious and contribute to the high incidence of organ dysfunction and infectious complications seen following injury . Regional hypoxia and depletion of intracellular energy stores occur in response to diminished microcirculatory blood flow, and these changes alter cellular signalling and result in the release of pro-inflammatory cytokines and prostanoids which mediate further suppression of immune function . Neutrophil priming serves to induce tissue damage in critical organ systems such as the lungs, heart, liver, and gut, further insulting the injured organism . Depression of antigen presentation and cytokine elaboration by macrophages and other antigen presenting cells effectively prevents a normal response from the acquired immune system, and lymphocyte-monocyte interactions are squelched . The resulting depression in cell mediated and humoral immunity renders the organism susceptible to microbial infection and contributes to the morbidity and mortality associated with nosocomial infections . Hormonal modulation of the immune response is highly evident following trauma and haemorrhage, and the preponderance of male morbidity associated with sepsis can be explained by the depression in immune function seen in males, but not females in the pro-oestrous state . Despite the multitude of changes induced by trauma and haemorrhage, experimental studies have revealed several promising pharmacologic interventions which may serve to blunt the effect of injury on the immune system, and render the host competent to withstand the bacterial and viral challenges responsible for so much of the late mortality following severe injury. Proteins, 1999 Jul 1, 36(1), 117 - 34 Conserved water molecules in a large family of microbial ribonucleases; Loris R et al.; We systematically analyzed the crystallographically determined water molecules of all known structures of RNase T1 and compared them to the ordered solvent in a large number of related microbial nucleases . To assess the crystallographers' impact on the interpretation of the solvent structure, we independently refined five validation structures from diffraction data derived from five isomorphous crystals of RNase T1 . We also compared the positions of water molecules found in 11 published isomorphous RNase T1 inhibitor complexes . These data suggest that the positions of most of the waters located on the surface of a protein and that are well-determined in the experimental electron density maps are determined primarily by crystal packing forces . Water molecules with less well-defined electron density are in general unique to one or a small number of crystal structures . Only a small number of the well-defined waters are found to be independent of the crystal environment . These waters have a low accessible surface area and B-factor, and tend to be conserved in the crystal structures of a number of evolutionary related ribonucleases as well . A single water molecule is found conserved in all known microbial ribonucleases. Anticancer Res, 1999 Mar-Apr, 19(2A), 1037 - 42 Thionins from Viscum album L: influence of the viscotoxins on the activation of granulocytes; Stein GM et al.; BACKGROUND: Extracts from European mistletoe (Viscum album L.) are applied in adjuvant cancer treatment, and some components, especially the mistletoe lectins (ML) have been immunologically characterised, but not the thionins, termed viscotoxins (VT) . MATERIALS AND METHODS: The influence of the VT on human granulocytes was studied by flow cytometry: E.coli co-stimulated respiratory burst by oxidation of dihydrorhodamine 123 to rhodamine 123 and phagocytosis by ingestion of FITC-labelled E.coli . RESULTS: VT (25 and 250 micrograms/ml), in contrast to ML, significantly enhanced phagocytosis and burst activity . VT-rich mistletoe extracts also exerted significant effects . In addition, E.coli-activated granulocytes positively stain with Annexin-V and propidium iodide only due to 250 micrograms/ml VT incubation, suggesting that at this concentration burst activity was induced by the physiological activity of granulocytes after microbial ingestion and also by cytotoxic effects . CONCLUSION: Viscotoxins exert yet unknown strong immunomodulatory effects on human granulocytes, which might be of benefit for tumour patients, in addition to their cytotoxic properties. Arch Dermatol Res, 1999 May, 291(5), 260 - 8 Stratum corneum thiol protease (SCTP): a novel cysteine protease of late epidermal differentiation; Watkinson A; Proteolytic enzymes play crucial roles in the formation of the stratum corneum barrier tissue and in its subsequent maturation . Despite this, the proteases involved in stratum corneum physiology are not well characterized . Hence, studies were performed to identify these proteolytic enzymes present in the peripheral layers of this tissue using a combination of tape stripping and zymography . Using this approach, a novel human cysteine protease was identified and characterized, and named stratum corneum thiol protease (SCTP) . Gelatin zymography revealed that SCTP is composed of two variants with apparent molecular weights of 34 and 35 kDa which do not correspond to any previously described stratum corneum protease . Mechanistically SCTP belongs to the cysteine proteinase class as shown by: (1) acid protease activity, (2) a requirement for mild reducing conditions, and (3) the specific inhibition of activity by E64 and Z-phe-ala-diazomethylketone . Further analysis using concanavalin A affinity chromatography demonstrated that the two 34 and 35 kDa variants are both glycoproteins, which, after removal of the oligosaccharide sidechains with the specific enzyme N-glycopeptidase F, reveal a single active core protease of 32 kDa . SCTP did not crossreact with antibodies raised against the lysosomal cysteine proteases cathepsins B, H or L, thereby distinguishing it from the classical cysteine cathepsins . Localization studies revealed that SCTP is present at all depths in the stratum corneum, thereby precluding microbial contamination as the enzyme source . Moreover, it was also present at all body sites investigated, except for the hyperkeratotic palmoplantar stratum corneum . SCTP was found to be a product of late differentiation in cultured human keratinocytes; the enzyme was synthesized by differentiated calcium-switched cells and secreted into the medium, whereas nondifferentiated basal keratinocytes did not produce this protease . Moreover, human fibroblast cultures did not produce the enzyme, suggesting that SCTP is not produced by the dermis and hence is epidermal specific . The function of SCTP is unknown, but the observed gelatinolytic activity coupled with its secretion into the medium by cultured keratinocytes indicates that physiologically it is responsible for the degradation of extracellular structural proteins . Furthermore, the optimal activity at acid pH suggests that it can function in the acidic environment of the stratum corneum . It remains to be elucidated whether this enzyme has a role in desquamation. J AOAC Int, 1999 May-Jun, 82(3), 648 - 55 Microbial ranking of porous packaging materials (exposure chamber method), ASTM method: collaborative study; Placencia AM et al.; A collaborative study involving 11 laboratories was conducted to measure the microbial barrier effectiveness of porous medical packaging . Two randomly cut samples from each of 6 commercially available porous materials and one positive and one negative control were tested by one operator in each of 11 laboratories . Microbial barrier effectiveness was measured in terms of logarithm reduction value (LRV), which reflects the log10 microbial penetration of the material being tested . The logarithm of the final concentration is subtracted from that of the initial concentration to obtain the LRV . Thus the higher the LRV, the better the barrier . Repeatability standard deviations ranged from 6.42 to 16.40; reproducibility standard deviations ranged from 15.50 to 22.70 . Materials B(53), C(50), D(CT), and E(45MF) differ significantly from the positive control . The microbial ranking of porous packaging materials (exposure chamber method), ASTM method, has been adopted First Action by AOAC INTERNATIONAL. Toxicol Sci, 1999 May, 49(1), 86 - 92 Factors contributing to the acute and subchronic adverse respiratory effects of machining fluid aerosols in guinea pigs; Gordon T et al.; Several physical, chemical, and microbial factors are potential contributors to the adverse pulmonary effects associated with occupational exposure to machining fluid aerosols . The present study examined the relative toxicity of 3 major classes of machining fluids (soluble, semi-synthetic, and synthetic) as well as that of unused (fresh) versus used (grab samples taken from manufacturing sites) machining fluids . Pulmonary function and changes in cellular and biochemical indices in bronchoalveolar lavage fluid were examined during and 24 h after exposure, respectively . Statistically significant differences in toxicity were observed in guinea pigs exposed for 3 h to respirable aerosols of unused machining fluids (semi-synthetic > soluble >> synthetic) . In addition, greater toxicity was observed in animals exposed to used, machining fluid aerosols compared to unused fluids . Moreover, within the used machining fluid types, significantly greater adverse effects were observed in animals exposed to poorly maintained fluids (i.e., heavy microbial contamination) versus well-maintained fluids . Changes in biochemical and cellular parameters in bronchoalveolar lavage fluid occurred after a single exposure to 5 mg/m3 of the poorly maintained used machining fluid aerosols . Changes in inflammation but not LDH and protein were observed in animals repeatedly exposed to semi-synthetic machining fluid aerosols . A statistically significant increase in lavage fluid neutrophils was observed in guinea pigs exposed to 5 mg/m3 used, semi-synthetic machining fluid aerosols for 4 weeks . In separate experiments, physicochemical properties of unused machining fluids were found to contribute to the production of adverse effects . Adjustment of the alkaline and hypotonic nature of the unused semi-synthetic machining fluid to isotonicity and pH 7 significantly reduced adverse effects . Together, these findings strongly suggest that multiple factors contribute to the adverse respiratory effects associated with occupational exposure to machining fluid aerosols. Mol Biotechnol, 1999 Feb, 11(1), 55 - 65 Mechanisms and assessment of lectin-mediated mitogenesis; Kilpatrick DC; The discovery of lectin-mediated mitogenesis by Nowell in 1960 stimulated interest in the properties of lectins while advancing knowledge of immunology . Although some lectins are polyclonal activators both in vitro and in vivo, others may display a broad range of activities toward human lymphocytes . Indeed, the same lectin (e.g., wheat germ agglutinin or Datura lectin) may be mitogenic, comitogenic, or antimitogenic, depending on the experimental conditions . An individual lectin may bind to several glycoproteins on the lymphocyte surface, resulting in interactions that may or may not be functionally relevant, and that may have opposing effects . Studies with lectins and with monoclonal antibodies (MAbs) have established that a surprisingly large variety of cell-surface molecules can influence the initiation and regulation of lymphocyte activation and proliferation . Interactions between lymphocytes and accessory cells are crucial; some signals are cell-mediated, but others depend on soluble cytokines . Mitogenic lectins presumably bind to the T-cell receptor complex and also promote a positive costimulatory signal leading to the synthesis of interleukin 2 and interleukin 2 receptors (IL-2R) . Nonmitogenic, comitogenic, and antimitogenic lectin activities also probably act via accessory molecules involved in costimulation . Plant lectin-animal lymphocyte interactions presumably have no physiological significance, but it is suggested that the former mimics microbial superantigens, which may function in the colonization of host cells . Mitogenic stimulation of lymphocytes can be assessed in several ways . The standard technique measures {3H}-thymidine incorporation into DNA, but nonradioactive procedures are also available. Int J Food Sci Nutr, 1998 Sep, 49(5), 343 - 52 Lipid content and fatty acid composition in foods commonly consumed by nursing Congolese women: incidences on their essential fatty acid intakes and breast milk fatty acids; Rocquelin G et al.; The fat content and fatty acid (FA) composition of nearly 40 foods, currently consumed by 102 nursing Congolese mothers living in Brazzaville, were determined to assess their impact on mothers' essential fatty acid (EFA) intakes and breast milk FA . Data on mothers' milk FA and dietary habits which allowed food selection were recently published (Rocquelin et al., 1998) . Most foods were locally produced . Food samples were collected at local markets, bleached if necessary to avoid microbial degradation, and stored at +4 degrees C or -20 degrees C . They were lyophilized upon their arrival in the laboratory before lipid analyses . FA composition of food lipids was determined by capillary gas chromatography . Staple diets included low-fat, high-carbohydrate foods (processed cassava roots, wheat bread) and high-polyunsaturated fatty acid (PUFA) foods: soybean oil (high in 18 : 2 n-6 and alpha-18 : 3 n-3), bushbutter (dacryodes edulis), peanuts, avocado (high in fat and 18 : 2 n-6), freshwater and salt-water fish (high in LC n-3 and/or n-6 PUFA), and leafy green vegetables (low in fat but very high in alpha-18 : 3 n-3) . Their frequent consumption by nursing mothers provided enough EFA to meet requirements due to lactation . It also explains why mothers' breast milk was rich in C8-C14 saturated FA (26% of total FA) and in n-6, n-3 PUFA (respectively 15.0% and 2.4% of total FA) highly profitable for breastfed infants' development . From this point of view, dietary habits of Congolese mothers have to be sustained for they are more adequate than most Western-type diets. Bioinformatics, 1999 May, 15(5), 362 - 9 Interpolated markov chains for eukaryotic promoter recognition; Ohler U et al.; MOTIVATION: We describe a new content-based approach for the detection of promoter regions of eukaryotic protein encoding genes . Our system is based on three interpolated Markov chains (IMCs) of different order which are trained on coding, non-coding and promoter sequences . It was recently shown that the interpolation of Markov chains leads to stable parameters and improves on the results in microbial gene finding (Salzberg et al., Nucleic Acids Res., 26, 544-548, 1998) . Here, we present new methods for an automated estimation of optimal interpolation parameters and show how the IMCs can be applied to detect promoters in contiguous DNA sequences . Our interpolation approach can also be employed to obtain a reliable scoring function for human coding DNA regions, and the trained models can easily be incorporated in the general framework for gene recognition systems . RESULTS: A 5-fold cross-validation evaluation of our IMC approach on a representative sequence set yielded a mean correlation coefficient of 0.84 (promoter versus coding sequences) and 0.53 (promoter versus non-coding sequences) . Applied to the task of eukaryotic promoter region identification in genomic DNA sequences, our classifier identifies 50% of the promoter regions in the sequences used in the most recent review and comparison by Fickett and Hatzigeorgiou ( Genome Res., 7, 861-878, 1997), while having a false-positive rate of 1/849 bp. Exp Cell Res, 1999 Jun 15, 249(2), 240 - 7 NA22598, a novel antitumor compound, reduces cyclin D1 levels, arrests cell cycle at G1 phase, and inhibits anchorage-independent growth of human tumor cells; Kawada M et al.; NA22598, a novel antitumor compound isolated from a microbial cultured broth, inhibited the growth of human colon cancer DLD-1 cells in suspension cultures (anchorage-independent growth) severalfold more strongly than in substratum-attached monolayer cultures . It arrested the cell cycle progression at early G1 phase under both these culture conditions . Rb phosphorylation, cyclin D1 expression, and cdk2 activation in G1 progression were all inhibited by NA22598, but the amounts of cdk2 and p27 were not affected . Among these effects the inhibition of cyclin D1 expression was most prominent, and NA22598 was found to inhibit the synthesis of cyclin D1 without affecting mRNA expression or protein degradation . p27 binding to cdk2 was more markedly increased in suspension cultures than in attached cultures by NA22598, but the compound had no effect on total p27 . Apparently, the decrease of cyclin D1 induced redistribution of p27 from the cyclin D1/cdk4 to the cyclin E/cdk2 complexes during G1 phase in the suspension cultures . Because p27 is upregulated during suspension culture, a greater amount of it was associated with cyclin E/cdk2, thus producing greater growth inhibition . An agent, like NA22598, which induces the downregulation of cyclin D1 might offer a new anticancer strategy . Ophthalmology, 1999 Jun, 106(6), 1166 - 70; discussion 1171 Risk factors in microbial keratitis leading to penetrating keratoplasty; Miedziak AI et al.; OBJECTIVE: To determine the characteristics of infectious corneal ulcers at the time of presentation to the cornea specialist associated with a favorable response to medical therapy versus a poor outcome manifested by the need for penetrating keratoplasty for therapy or visual rehabilitation . DESIGN: Retrospective, case-control study . PARTICIPANTS: A total of 162 patient records were reviewed, including the study group of 30 patients and the control group of 132 patients . INTERVENTION: A retrospective review of all cases of microbial keratitis presenting to the Cornea Service between January 1, 1989 and December 31, 1995 was conducted . The cases were divided into two groups . The study group consisted of patients with microbial keratitis who failed medical therapy and required penetrating keratoplasty . The control group included patients with infectious ulcers who responded to medical therapy alone . MAIN OUTCOME MEASURES: The influence of demographics, medical and ocular history, delay in presentation to the primary ophthalmologist or the corneal specialist, topical medications, and contact lens usage were compared . Visual acuity and ulcer characteristics were recorded . The statistical significance was evaluated by the chi-square test for independence and multiple logistic regression . RESULTS: Older age (P=0.001), delay in referral to the corneal specialist (P<0.03), and treatment with topical steroids prior to presentation (P<0.0001) were statistically significant factors associated with the need for penetrating keratoplasty . Steroid use and the delay in referral were correlated . A past history of ocular surgery (P=0.01), poor visual acuity at presentation (P<0.001), and ulcer characteristics, including central location (P<0.0001), large size (P<0.0001), presence of perforation or descemetocele (P<0.0001), limbal involvement (P<0.0001), and hypopyon (P=0.05), were all associated with the need for penetrating keratoplasty . CONCLUSIONS: Older age, delay in referral to the corneal specialist, topical steroid treatment, past ocular surgery, poor vision at presentation, large size, and central location of the ulcer are risk factors for poor outcome of microbial keratitis, as indicated by the need for penetrating keratoplasty. Phytochemistry, 1999 May, 51(2), 257 - 61 Microbial metabolism of artemisitene; Orabi KY et al.; Studies on the microbial transformation of the sesquiterpene endoperoxide artemisitene have revealed that artemisitene was metabolized by Aspergillus niger (NRRL 599) to yield 11-epi-artemisinin, 9 beta-hydroxydeoxy-11-epi-artemisinin and 9 beta-hydroxy-11-epi-artemisinnin . These metabolites were characterized on the basis of their spectral data. Science, 1999 Jun 11, 284(5421), 1835 - 7 The nature of the principal type 1 interferon-producing cells in human blood; Siegal FP et al.; Interferons (IFNs) are the most important cytokines in antiviral immune responses . "Natural IFN-producing cells" (IPCs) in human blood express CD4 and major histocompatibility complex class II proteins, but have not been isolated and further characterized because of their rarity, rapid apoptosis, and lack of lineage markers . Purified IPCs are here shown to be the CD4(+)CD11c- type 2 dendritic cell precursors (pDC2s), which produce 200 to 1000 times more IFN than other blood cells after microbial challenge . pDC2s are thus an effector cell type of the immune system, critical for antiviral and antitumor immune responses. Curr Opin Biotechnol, 1999 Jun, 10(3), 225 - 9 16S rDNA methods in soil microbiology O'Donnell AG, Gorres HE. With the introduction of molecular methods, the past decade has seen renewed interest in soil microbiology . New and exciting molecular technologies and the promise of finally opening the microbial black box in soil drive much of this interest . Although these pioneering studies have added much to our knowledge of microbial diversity in soils, it is debatable whether they have as yet advanced our understanding of the relationship between this diversity and soil processes . Hopefully, over the next few years, the knowledge gained from molecular studies will provide a better understanding of microbial communities in soils and lead ultimately to improvements in land management and to the exploitation of the genetic resources of soil. Curr Opin Biotechnol, 1999 Jun, 10(3), 220 - 4 Molecular approaches to the study of aquatic microbial communities MacGregor BJ. Molecular methods for studying microbial communities are still under development . Enormous sequence catalogues can be collected; they must now be related to microbial activities . Messenger RNA detection, fluorescent in situ hybridization, cell sorting, and oligonucleotide array technology are currently being explored . Biases are associated with all these methods, but combined approaches offer checks and balances. N Y State Dent J, 1999 Apr, 65(4), 32 - 7 Current and future approaches for diagnosis of periodontal diseases; Papapanou PN et al.; Our understanding of the etiology and pathogenesis of periodontal diseases has grown in the recent past, and new findings have led to advances in patient management . This article summarizes important new knowledge and offers a description of traditional and novel diagnostic approaches . These include clinical and radiographic assessments of the periodontal tissues, evaluation of the microbial challenge and the host response, and certain elements of the host genotype that may confer susceptibility to destructive periodontal diseases. Hum Cell, 1998 Dec, 11(4), 201 - 6 {Control of tumor-related angiogenesis}; Oikawa T; Tumor-related angiogenes is expected to become an important target for improving treatment of cancer, because it plays key roles in tumor growth, invasion and metastasis . We considered that the successful development of such angiostatic treatment depended entirely upon the development of useful anti-angiogenic agents, and attempted to find novel angiogenesis inhibitors by using three in vivo assays, based on an idea of ours . As a result we have demonstrated that different types of agents with low molecular weight including microbial metabolites, cell differentiation modulators like retinoids and steroids, exhibit anti-angiogenic activity, anti-metastatic activity and/or antitumor activity . Taken these findings, an ideal anti-angiogenic agent is discussed. Am J Physiol, 1999 Jun, 276(6 Pt 1), C1231 - 42 Molecular mechanisms of cytoadherence in malaria; Ho M et al.; Microbial pathogens subvert host adhesion molecules to disseminate or to enter host cells to promote their own survival . One such subversion is the cytoadherence of Plasmodium falciparum-infected erythrocytes (IRBC) to vascular endothelium, which protects the parasite from being removed by the spleen . The process results in microcirculatory obstruction and subsequent hypoxia, metabolic disturbances, and multiorgan failure, which are detrimental to the host . Understanding the molecular events involved in these adhesive interactions is therefore critical both in terms of pathogenesis and implications for therapeutic intervention . Under physiological flow conditions, cytoadherence occurs in a stepwise fashion through parasite ligands expressed on the surface of IRBC and the endothelial receptors CD36, intercellular adhesion molecule-1 (ICAM-1), P-selectin, and vascular adhesion molecule-1 . Moreover, rolling on ICAM-1 and P-selectin increases subsequent adhesion to CD36, indicating that receptors can act synergistically . Cytoadherence may activate intracellular signaling pathways in both endothelial cells and IRBC, leading to gene expression of mediators such as cytokines, which could modify the outcome of the infection. Eur Respir J, 1999 Apr, 13(4), 835 - 8 Antibiotic use in patients admitted with acute exacerbations of chronic obstructive pulmonary disease; Smith JA et al.; The objective of this report was to document the pattern of initial antibiotic prescribing in acute exacerbations of chronic obstructive pulmonary disease (COPD) in a hospital setting . All episodes of acute exacerbation of COPD, as diagnosed by the admitting doctor, in one hospital in the period January to May 1996, were identified . Case notes were reviewed retrospectively . Cases of radiographic pneumonia, bronchiectasis and incorrectly coded admissions were excluded . Symptoms, microbial cultures and initial antibiotic therapies were recorded . One hundred and fifty-nine patient episodes were identified; 40 were excluded yielding a sample of 119 . Nineteen case notes were unavailable leaving a sample of 100 (84%) episodes . Eighty were treated with antibiotics on admission; amoxycillin was the most frequently prescribed, in 46 (58%) episodes . Of the antibiotic treated group, 42 (53%) patients were given dual therapy, most commonly a macrolide antibiotic with either amoxycillin or a cephalosporin . Intravenous treatment was used in 22 (28%) cases . The duration of intravenous treatment was >48 h in 12 (15%) cases . A total of 76 sputum samples were analysed from 55 patient episodes: 34 (45%) were culture positive . In 15 (27%) patient episodes, antibiotic therapy was changed or instituted on the basis of culture results . These data suggest that antibiotic treatment is not optimal, with overuse of antibiotics, especially intravenous and dual therapy. Zentralbl Bakteriol, 1999 Apr, 289(2), 147 - 54 Cold-induced augmentation of I blood group antigen interactions with galactophilic lectins; Sudakevitz D et al.; The I antigen appears on human cells in the postnatal period, by addition of N-acetyllactosamine (beta 1-6) branching to the fetal i antigen structure, which is specified by linear oligo N-acetyllactosamine (beta 1-3) chain . Concurrently with the I antigen appearance on adult human erythrocytes most human sera exhibit low levels of anti-I agglutinins . These antibodies induce hemagglutination mainly at low temperatures (4 degrees C) and scantly at body temperature . Therefore they were named "cold agglutinins" . We have used these antibodies and several hemagglutinating galactophilic animal, plant, and microbial lectins that also react with the I antigen, to study whether the cold-favored agglutination of the I antigen-bearing cells is a peculiar property of the anti-I antibodies or a special trait of that antigen . It has been found that the interactions of all of the examined lectins, irrespective of their source, with the adult human erythrocytes significantly increased at 4 degrees C, in contrast to those of the same cells with diverse I-insensitive antibodies and lectins, which were significantly higher at room temperature. Int Immunol, 1999 Jun, 11(6), 899 - 906 Expansion of neonatal tolerance to self in adult life: I . The role of a bacterial adjuvant in tolerance spread; Grabie N et al.; T cell neonatal tolerance to self evolves perturbation of the Th1/Th2 balance towards Th2-type self-specific T cells . In the current study we have demonstrated that a tolerant state could be extended to another encephalitogenic determinant only if the neonatally tolerizing determinant was co-administered in adult life with an emulsion of Mycobacterium tuberculosis (i.e . complete Freund's adjuvant) . The mechanisms underlying tolerance elicitation and expansion were then explored by an in vitro system in which indirect suppression could be measured . Addition of a tolerizing epitope to splenic T cells from neonatally tolerized animals induced a marked suppression of the anti-MT response . This response could be restored by neutralizing antibodies to IL-4 . In contrast, the neutralizing antibodies to IL-4 had no affect on the response of these cells to the tolerizing determinant . These findings are highly significant not only because they explore the important role of microbial antigens in neonatal tolerance, but also because they distinguish, for the first time, between tolerizing and tolerized T cells. J Allergy Clin Immunol, 1999 Jun, 103(6), 1153 - 7 A cross-sectional survey of sensitization to Aspergillus oryzae-derived lactase in pharmaceutical workers; Bernstein JA et al.; BACKGROUND: The presence of IgE-mediated occupational respiratory sensitization to microbial enzymes has been well documented in a variety of industries . Aspergillus oryzae -derived lactase is used as a dietary aid for patients with lactose intolerance . OBJECTIVE: In 1993, a cross-sectional survey of 94 pharmaceutical workers exposed to lactase for a mean duration of 23 months and 24 nonexposed recently hired employees was initiated to identify lactase-sensitized workers and potential risk factors that could be used in making recommendations for preventing future cases of lactase sensitization . METHODS: The survey included a physician-administered questionnaire, skin prick testing to lactase enzyme and a panel of common aeroallergens, and spirometry . RESULTS: Twenty-seven of 94 lactase-exposed workers (29%) had positive skin test responses to lactase . These workers were 9 times more likely to have upper or lower respiratory symptoms compared with workers with negative skin test responses . Atopic workers were 4 times more likely to have lactase skin sensitivity than nonatopic workers . However, atopy was not a risk factor for the development of upper and/or lower respiratory symptoms . Lactase skin reactivity was not observed in the 24 nonexposed employees . CONCLUSION: This cross-sectional survey revealed that atopic workers were more likely to have lactase sensitization and that lactase-sensitized workers were more likely to have upper and/or lower respiratory symptoms, but atopy was not a risk factor for upper or lower respiratory symptoms