Vaccine, 1996 Aug, 14(12), 1149 - 53 Oral application of romurtide, a synthetic muramyl dipeptide derivative, stimulates nonspecific resistance to microbial infections and hematopoiesis in mice; Namba K et al.; Romurtide given orally enhanced the nonspecific resistance against microbial infections and hematopoiesis up to the levels achieved by subcutaneous (s.c.) injection of the compound in mice . Oral romurtide conferred protection and, in consequence, enhanced therapeutic efficacy of antibiotics against systemic infections in mice . The leukocytosis followed by the elevations of colony stimulating activity in serum and the colony forming unit of granulocyte-macrophage (c.f.u.-GM) in femoral bone marrow was observed as successive event in mice treated orally with romurtide . To obtain a comparable potency to s.c . injection of the compound at a dose of 0.1 mg per mouse, oral application required doses of 3 and 10 mg per mouse for stimulating the nonspecific resistance to infection and hematopoiesis, respectively. Comput Appl Biosci, 1996 Aug, 12(4), 353 - 6 Software for microbial fingerprinting by means of the infrared spectra; Maradona MP; Two computer programs were designed for helping in library handling and microbial identification by means of their infrared spectra . The program 'Transform' runs in the IR Data Manager environment and produces ASCII files containing transformed data from infrared absorbance spectra . The program 'WinSpectra' is written in Visual Basic v . 3.0 . It imports the ASCII files created with 'Transform', and is able to handle, analyse and identify them by their mathematical comparison with a library of microbial spectra . Both programs run in graphical, menu-driven interface (GEM and MS Windows, for IBM-PC compatibles, with extensive on-line help. J Oral Pathol Med, 1996 Aug, 25(7), 360 - 6 Salivary protein composition in epileptic patients on different medications; Henskens YM et al.; Several salivary proteins were assayed in saliva from epileptic patients who were using different anti-epileptic drugs, viz, phenytoin, valproate and carbamazepine, and were compared with levels in unmedicated healthy control subjects . Flow rate and pH of the patient groups were not different from the controls . In all patient groups the specific amylase activity was increased up to twofold . In the phenytoin group only, the salivary IgA concentration was strongly reduced . Levels of salivary cystatin C were similar among all patient groups studied, and were not different from those of the control group . In contrast, the absolute and relative concentrations of cystatin S were diminished, particularly in patients using either valproate or phenytoin . These data suggest that use of anti-epileptic drugs over long periods may result in decreased levels of several salivary proteins such as sIgA and cystatins, which are involved in the protection of the oral cavity against microbial infections. J Dairy Sci, 1996 Aug, 79(8), 1410 - 24 Effects of dietary fat with or without nicotinic acid on nutrient flow to the duodenum of dairy cows; Christensen RA et al.; Four Holstein cows, fitted with ruminal and duodenal cannulas, were utilized in a 4 x 4 Latin square design to investigate the effects of supplementing nicotinic acid to diets that contained 35% alfalfa haylage, 15% corn silage, and either 50% of a low fat concentrate or 10% whole raw soybeans and 40% of a high fat concentrate containing tallow . Treatments in a 2 x 2 arrangement were 1) low fat, no supplemental nicotinic acid; 2) low fat, 12 g/d of nicotinic acid; 3) high fat, no supplemental nicotinic acid; and 4) high fat, 12 g/d of nicotinic acid . The DMI and OM apparently and truly digested in the rumen and apparently digested postruminally were not different among treatments . Addition of fat to the diet decreased the concentration of total VFA in ruminal fluid but did not alter the molar proportions of any of the VFA; supplementation of nicotinic acid tended to decrease the molar proportion of acetate . Amounts of NAN, microbial N, nonammonia nonmicrobial N, and AA that flowed to the duodenum were similar among diets . The concentration of urea N in plasma decreased, and concentrations of cholesterol and triglycerides increased, when cows were fed supplemental fat . Milk composition and production of milk, 4% FCM, and milk components were not altered by addition of fat or nicotinic acid to the diet . Supplementation of fat or nicotinic acid to diets of dairy cows was not beneficial in this experiment. J Hematother, 1996 Aug, 5(4), 351 - 7 Granulocytes harvested following G-CSF-enhanced leukocyte recovery retain their functional capacity during in vitro culture for 72 hours; Murea S et al.; The purpose of this study was to compare two different in vitro culture conditions for the preservation of human granulocytes . These cells could be used in patients with severe neutropenia following cytotoxic chemotherapy if the functional capacity was retained, and autologous transfusions of granulocytes would circumvent the risk of alloimmunization . Granulocytes were obtained from the peripheral blood of healthy donors and patients with hematologic malignancies who received cytotoxic chemotherapy supported by recombinant human granulocyte colony-stimulating factor (R-metHuG-CSF, 300 micrograms/day, s.c.) . Granulocytes were either cultured for 72 h at 4 degrees C in the presence of 100 ng/ml G-CSF or cryopreserved at -196 degrees C . The viability, surface antigen expression, and function of the granulocytes were assessed . Since effective microbial killing involves the attachment of granulocytes to blood vessel walls, transmigration into tissues, chemotaxis, and phagocytosis, the surface expression of the adhesion molecules LFA-1 (CD11a/CD18) and gp 150,95 (CD11c/CD18) was measured . In addition, the IgG receptors Fc gamma RI (CD64), Fc gamma RII (CD32), and Fc gamma RIII (CD16), as well as the complement receptor CR3 (CD11b/CD18), were assessed . Dynamic superoxide anion release served as a measure of the metabolic pathway of the oxidative burst after f-Met-Leu-Phe (fMLP) and phorbol-12-myristate-13-acetate (PMA) stimulation . Substantial differences in the preservation of granulocyte integrity and function were observed between the two storage conditions . Cryopreservation abolished reactivity to extracellular stimuli and severely affected the cell phenotype . On the other hand, functional activity could be maintained for up to 72 h when in vivo primed granulocytes of patients were incubated at 4 degrees C in the presence of G-CSF . This storage modality may permit the use of granulocyte autotransfusion to reduce the risk of neutropenic fever. Am J Infect Control, 1996 Aug, 24(4), 304 - 9 Advantages and limitations of automatic flexible endoscope reprocessors; Muscarella LF; The advantages and limitations of automatic flexible endoscope reprocessors (AFERs) are presented . These medical devices can increase patient safety and minimize personnel exposure to the liquid chemical germicide and its vapors . Some models feature a printer to document important reprocessing parameters, an in-line tap water filtration system to reduce the likelihood of endoscope (and patient) contamination, and a heater to satisfy the elevated temperature requirements of some liquid chemical germicide labels . Although they offer significant advantages compared with manual reprocessing, AFERs have their limitations . Current models do not automate every endoscope reprocessing step and are not typically designed to reprocess every endoscope channel . Some AFER models may have to be periodically decontaminated to ensure that their internal components do not support microbial colonization . Moreover, their cost per cycle can be considerably more than manual reprocessing without necessarily reducing overall reprocessing time . Its limitations notwithstanding, an AFER is likely to be an asset in a busy endoscopy center . In addition to comparing automated and manual reprocessing, this article addresses important reprocessing issues and purchasing concerns. Eur J Gastroenterol Hepatol, 1996 Aug, 8(8), 829 - 33 Duodenal ulcer disease; Tytgat GN; An overview is given of the pathogenic mechanisms involved in Helicobacter pylori-associated duodenal and gastric ulceration . Special attention is given to the role of microbial virulence factors, the effects on gastric acid secretion and the development of 'gastric type' metaplasia in the duodenal bulb . Cure of the infection essentially eliminates ulcer recurrence . Cure also prevents ulcer-related bleeding . Antibiotic regimens providing a reliable high cure rate are briefly discussed . Finally the questions about ulcer disease which remain unanswered are summarized. J Anim Sci, 1996 Aug, 74(8), 1973 - 81 Barley varieties for finishing steers: II . Ruminal characteristics and rate, site, and extent of digestion; Boss DL et al.; Four ruminally and abomasally cannulated steers were used to evaluate the effects of barely variety on rate, site, and extent of digestion of high-concentrate diets . The treatments compared were 1) corn, 2) Gunhilde barley (GUN), Harrington barley (HAR), and 4) Medallion barley (MED) . Diets were balanced to be isocaloric and isonitrogenous . Ruminal OM digestion was greater (P = .04) in steers fed corn than in those fed GUN, HAR, and MED (61.9 vs average 53.7%) . No differences (P > .10) were seen in ruminal starch digestion (average 92.8%) or in starch flow to the abomasum (average 199 g/d) between diets . Total tract digestion of starch was greater (P = .09) in steers fed barley than in those fed corn (average 98.6 vs 95.7%) . Total and nonammonia N presented to the abomasum were greater (P < .05) for steers fed HAR and GUN than for those fed MED and corn . Microbial N flow was lowest (P = .01) in corn-fed steers, highest in steers fed GUN and HAR, and intermediate in steers fed MED . Microbial efficiency was 59% greater (P = .03) in steers fed barley than in steers consuming corn . Ruminal acetate: propionate was lower (P = .002) in steers fed corn and HAR than in those fed GUN and MED . Compared to GUN, HAR, and MED barleys, corn had a lower (P < .03) rate (-.11 vs average -.47) and extent (15 h; 70.3 vs average 98.1%) of in situ starch disappearance . Differences in digestive characteristics found between barley varieties may contribute to differences in animal performance. J Anim Sci, 1996 Aug, 74(8), 1873 - 87 Mathematical modeling of digestion and nutrient absorption in pigs; Bastianelli D et al.; A simple simulation model of digestion and absorption in pigs was developed . The structure of the model is a set of four anatomical compartments for DM: stomach, two portions of small intestine, and the large intestine . In each of these anatomical compartments, subcompartments correspond to the major biochemical components of feed and their products of degradation . The major degradation and absorption events are considered as well as the effect of microbial activity in the large intestine . The total number of compartments is 44 . The numerical integration with a time step of 1 min allows prediction of kinetic features of digestion phenomena such as absorption patterns and transit flows . First validation of the model shows that the global dynamic behavior of the model is realistic and promising . However, some additional factors must be considered for improved accuracy, in particular the susceptibility of the feed components to enzymatic degradation . The outputs of such a model could be used as inputs for metabolic or growth models running with time steps smaller than the 24-h basis often used in nutrition. Cardiol Clin, 1996 Aug, 14(3), 405 - 36 Advances in medical and antibiotic management of infective endocarditis; Kubak BM et al.; Infective endocarditis remains a serious medical problem despite advancements in laboratory detection, echocardiographic techniques, and newer antibiotic agents . This article summarizes the microbial agents in infective endocarditis, in addition to developments in medical and antibiotic management. Poult Sci, 1996 Aug, 75(8), 979 - 90 Effect of microbial phytase on nitrogen and amino acid digestibility and nitrogen retention of turkey poults fed corn-soybean meal diets; Yi Z et al.; The effect of microbial phytase on N and amino acid (AA) digestibility and N retention was investigated in a 29-d trial using 480 Nicholas Large White Turkey female poults fed corn-soybean meal diets . A 2 x 2 x 2 factorial arrangement of treatments was used with 0.45 and 0.60% nonphytate P (nP), 22.5 and 28.0% CP, and 0 and 750 U of microbial phytase/kg of diet . At 0.45% nP, adding phytase to either 22.5 or 28.0% CP diets increased BW gain (P < 0.01), and percentage (P < 0.01) and weight (P < 0.10) of toe ash; at 0.60% nP, the magnitude of the effect of phytase was less (P > 0.10) than observed for 0.45% nP and inconsistent . Apparent and true ileal digestibility of N and AA was estimated by using chromic oxide as an indicator at Day 24 . At 0.45% nP, adding phytase to 22.5% CP diets tended to improve the apparent and true ileal digestibility of N and AA, except cysteine or methionine; adding phytase to 28.0% CP diets increased the digestibility of N and most of the AA (P < 0.001 to 0.10) . At 0.60% nP, adding phytase to 22.5% CP diets increased the apparent and true ileal digestibility of N and all the AA (P < 0.001 to 0.10), but did not change digestibilities at 28.0% CP diets . Adding phytase also increased (P < 0.001 to 0.10) apparent ileal digestibility of DM and P at 0.45% nP for both CP diets, but only for 22.5% CP diets at 0.60% nP . The total excreta were collected at Day 27 to 29 . Adding phytase to 0.45% nP diets increased apparent utilization of DM (P < 0.01 to 0.10) and retention of N (P < 0.05 to 0.10) at both CP levels; retention of P was only increased (P < 0.10) at 22.5% CP . At 0.60% nP, adding phytase increased utilization of DM (P < 0.05) and retention of N (P < 0.10) only at 22.5% CP; P retention was not affected . In summary, microbial phytase enhanced growth performance, toe ash, ileal N and AA digestibility, and apparent N and P retention. J Antibiot (Tokyo), 1996 Aug, 49(8), 802 - 10 Microbial conversion of EM574 and EM523, gastrointestinal motor stimulating agents; Funabashi Y et al.; EM574 exerts gastrointestinal motor stimulating (GMS) activity even after being converted to its metabolites P1 and P2 in dogs . These metabolites were isolated from dog liver using a series of chromatographic procedures . Their structures were determined to be the 15- and 14-hydroxyl derivatives of EM574, respectively, by spectral analysis . Large scale preparation by microbial transformation was investigated for further evaluation of the metabolites, because the amounts obtained by oxidation with dog liver homogenate were limited . Three strains of actinomycetes, Amycolatopsis tolypophorus IFO 13151, Dactylosporangium variesporum IFO 14104 and Nocardia capreola IFO 12847, were found to have the aiming oxidative potency . HPLC analysis of the crude extracts from these three cultures showed that the bioactive metabolites, EM574 P1 and P2 were produced . They were isolated from the culture broth with the other bioactive products EM574 P3 and P4 . These bioactive products were prepared by large scale cultivation . EM574 P3 and P4 showed GMS activity comparable to that of EM574 P1 and P2 . The structures of EM574 P3 and P4 were elucidated by spectral analysis and found to be the 3"-O-demethyl derivatives of EM574 P2 and EM574, respectively . Moreover, the absolute configuration at the C14 position of P2 was determined to be R by spectral analysis of the 6-membered cyclic carbonate of EM574 P2. J Antibiot (Tokyo), 1996 Aug, 49(8), 781 - 7 Microbial biotransformation products of cyclosporin A; Kuhnt M et al.; In order to mimic the human metabolic pathway of cyclosporin A (CyA) a total of 28 bacterial and 72 fungal strains was screened for their ability to transform CyA . Among 3 bacteria and 11 fungi, which produced the main human metabolite OL-17 {eta HyMeBmt1}CyA, Actinoplanes sp . (ATCC 53771) achieved the best transformation rate (5.4%) . Furthermore, the two N-demethylated minor products {Leu4}CyA (3.2%) and {Leu9}CyA (4.7%) were isolated, both known as minor natural metabolites and the first one also as a human biotransformation product . Microbial conversion of CyA using the actinomycete Sebekia benihana (NRRL 11111) yielded {gamma HyMeLeu4}CyA (35%), {gamma HyLeu4}CyA (4.5%) and {gamma HyMeLeu4, gamma HyMeLeu6}CyA (8.6%) . The structures of these derivatives correspond with those of the human metabolic pathway . The related compounds {Nva2}CyA (CyG) and {D-MeSer3}CyA were similarly converted to the corresponding 4-gamma-hydroxylated analogues . None of the biotransformation products showed a better immunosuppressive effect than CyA, although in various cases the cyclophilin binding affinity was comparable to that of CyA. Risk Anal, 1996 Aug, 16(4), 549 - 63 Quantifying water pathogen risk in an epidemiological framework; Eisenberg JN et al.; Traditionally, microbial risk assessors have used point estimates to evaluate the probability that an individual will become infected . We developed a quantitative approach that shifts the risk characterization perspective from point estimate to distributional estimate, and from individual to population . To this end, we first designed and implemented a dynamic model that tracks traditional epidemiological variables such as the number of susceptible, infected, diseased, and immune, and environmental variables such as pathogen density . Second, we used a simulation methodology that explicitly acknowledges the uncertainty and variability associated with the data . Specifically, the approach consists of assigning probability distributions to each parameter, sampling from these distributions for Monte Carlo simulations, and using a binary classification to assess the output of each simulation . A case study is presented that explores the uncertainties in assessing the risk of giardiasis when swimming in a recreational impoundment using reclaimed water . Using literature-based information to assign parameters ranges, our analysis demonstrated that the parameter describing the shedding of pathogens by infected swimmers was the factor that contributed most to the uncertainty in risk . The importance of other parameters was dependent on reducing the a priori range of this shedding parameter . By constraining the shedding parameter to its lower subrange, treatment efficiency was the parameter most important in predicting whether a simulation resulted in prevalences above or below non outbreak levels . Whereas parameters associated with human exposure were important when the shedding parameter was constrained to a higher subrange . This Monte Carlo simulation technique identified conditions in which outbreaks and/or nonoutbreaks are likely and identified the parameters that most contributed to the uncertainty associated with a risk prediction. Br J Nutr, 1996 Aug, 76(2), 231 - 48 The fate of absorbed and exogenous ammonia as influenced by forage or forage-concentrate diets in growing sheep; Lobley GE et al.; Changes in splanchnic energy and N metabolism were studied in sheep, prepared with vascular catheters across the portal-drained viscera (PDV) and the liver, and maintained on supramaintenance intakes of either gross or grass + barley pellets . The animals were challenged, on both diets, with 4 d intramesenteric vein infusions of NH4Cl (25 mumol/min) plus NH4HCO3 (at either 0 or 125 mumol/min) . On the final day of each treatment the natural abundance NH4Cl was replaced with 15NH4Cl over a 10 h infusion while over the same period {1-13C}leucine was infused via a jugular vein . Measurements were made of blood flow plus mass transfers of NH3, urea, free amino acids and O2 across the PDV and liver . Enrichments of {14N15N}urea and {15N15N}urea plus {15N}glutamine, aspartate and glutamate were also monitored . Whole-body urea flux was determined by infusion of {14C}urea . At the end of the study the animals were infused for 3 h with 15nH4Cl, killed and liver samples assayed for intracellular free amino acid enrichments and concentrations . Blood flows across the splanchnic region were unaffected by either diet or level of ammonium salt infusion . At the lower ammonium salt infusion there was a trend for greater absorption of NH3 across the PDV (P < 0.10) with grass + barley than with the grass diet, while removal of urea was unaltered . At the higher ammonium salt infusions there was a significantly greater appearance of NH3 across the PDV and this exceeded the extra infused . Urea-N removal, however, was also elevated and by more than that required to account for the additional NH3 . The PDV contributed 19-28% to whole-body O2 consumption and the liver 23-32% . Hepatic extraction of absorbed NH3 was complete on all treatments and systemic pH remained constant . The fractions of urea-N apparently derived from NH3 were similar on the grass (0.50-0.64) and grass + barley (0.64-0.67) diets . Hepatic production of urea agreed well with urea flux measurements . Between the two levels of ammonium salt infusion and within diets the additional NH3 removed across the PDV was accounted for by the increased urea-N production . The {14N15N}:{15N15N} ratio of the urea produced was 97:3, while the enrichment of hepatic intracellular free aspartate was lower than that of {14N15N}urea . Glutamine enrichments were 0.23-0.37 those of {14N15N}urea, indicating a minor role for those hepatocytes (probably perivenous) which contain glutamine synthetase (EC 6.3.1.2) . Leucine kinetics, either for the whole body or splanchnic tissues, were not different between diets or level of ammonium salt infusion, except for oxidation which was less on the grass + barley ration . Amino acid concentrations were lower on the grass + barley diet but net PDV absorptions were similar . The pattern of essential amino acids absorbed into the PDV showed good agreement with the published composition of mixed rumen microbial protein . Fractional disappearances of absorbed free essential amino acids across the liver varied from 0.4 (branched chains) to near unity (histidine, phenylalanine). Nahrung, 1996 Aug, 40(4), 194 - 200 Prolongation of the keeping quality of Mozzarella cheese by treatment with sorbate; Aly ME; An attempt was carried out for prolongation of the shelf-life of Mozzarella cheese by incorporation of potassium sorbate into the cheese . Three manufacturing techniques were used: a) addition of potassium sorbate to kneading water (at level of 6%), b) addition during brine salting (at level of 0.5%) and c) dipping the cheese into potassium sorbate solution (6%) directly prior to packaging . Control cheese was made without potassium sorbate treatment . The resulting cheeses were divided into two portions, one of which was contaminated with Penicillium roqueforti and then packaged, while the second one was packaged without contamination . Both were stored at refrigerator (5 +/- 1 degree C) temperature and analysed periodically until spoilage . The results showed that treatment with potassium sorbate did not affect the organoleptic properties of the cheeses, except that a slight objectionable bitter flavour was observed in fresh cheeses treated with sorbate using the techniques of dipping or in brine salting then it was disappeared during storage . However, the overall acceptabilities of the sorbate-treated cheese were increased up to 10 weeks of storage compared with 4 weeks for untreated cheeses . Treatment with potassium sorbate in kneading water or brine appeared to be more effective than dipping . Addition of potassium sorbate inhibited microbial growth, especially that of moulds and yeasts . The sorbate-treated cheeses had higher moisture, pH values and lower acidity than the control . Fat, salt and total nitrogen were unaffected during storage . Levels of soluble N, non-protein N and total volatile fatty acids in sorbate-treated cheeses were slightly higher than in the control . Furthermore, addition of potassium sorbate increased the meltability and improved the fat leakage of Mozzarella cheese. Curr Opin Immunol, 1996 Aug, 8(4), 510 - 6 T-cell recognition of non-peptide antigens; Porcelli SA et al.; Studies of two distinct human T-cell systems have provided the exciting finding that T cells are able to recognize non-peptide antigens: gammadelta T cells have been shown to recognize isopentenyl pyrophosphate and related structures and human CD1 has been shown to present microbial lipids and lipoglycans such as mycolic acids and lipoarabinomannan to T cells . T-cell responses to these non-peptide antigens should provide a strategic target for immunologic intervention in infectious disease. Am J Trop Med Hyg, 1996 Aug, 55(2), 165 - 73 Risk of human exposure to vector ticks (Acari: Ixodidae) in a heavily used recreational area in northern California; Lane RS; The risk of humans encountering vector ticks along hiking trails or in picnic grounds in a Lyme disease-endemic area was evaluated in a multipurpose recreational area (Tilden Regional Park) in the populous San Francisco Bay region of California . Four hillside hiking trails (two high-use, two low-use) were sampled by dragging and walking through low vegetation biweekly for one year; four heavily used picnic areas were sampled concurrently by dragging . Adults of three human-biting ticks were enumerated (n = 1,911) along all trials: Dermacentor occidentalis (63.6% of total), Ixodes pacificus (26.2%), and D . variabilis (0.2%) . Subadults (n = 1,669) of D . occidentalis (0.06% of total) and two nonhuman-biters, D . albipictus (70.3%) and Haemaphysalis leporispalustris (29.7%), also were collected . Dragging yielded many more adult ticks than walking year-round for all trials . These methods were significantly correlated during periods of peak tick abundance, but the associations were not sufficiently strong or consistent to allow prediction of captures for either method based on the other . Adult ticks were distributed largely in clusters along the uphill sides of trails . Several adult ticks collected adjacent to trails were found to contain spirochetes identified with polyclonal antibodies as Borrelia burgdorferi (D . occidentalis, 0 of 861; D . variabilis, 2 of 126 {1.6%}; I . pacificus, 1 of 609 {0.2%}) . Picnic areas produced low numbers of adult D . occidentalis and I . pacificus, which prohibited testing them for spirochetes . Two measures for evaluating risk were calculated, the encounter distance (= mean number of meters traveled before encountering a vector tick by either dragging or walking) and the mean number of spirochete-infected ticks encountered by these methods per kilometer of trial . These measures revealed that the risk of exposure to spirochete-infected adult ticks along trails was low year-round irrespective of usage, and risk was even lower in picnic areas . Future studies evaluating human exposure to vector ticks in recreational areas should incorporate, whenever possible, testing for multiple pathogens because most ixodid ticks that commonly bite people in the United States are capable of transmitting two or more microbial disease agents. Microbiology, 1996 Aug, 142 ( Pt 8), 1973 - 82 Evidence for a novel class of microbial 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase in Streptomyces coelicolor A3(2), Streptomyces rimosus and Neurospora crassa; Walker GE et al.; The tryptophan-sensitive 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthases from Streptomyces coelicolor A3(2), Streptomyces rimosus and Neurospora crassa have been purified to homogeneity . All three enzymes have a subunit Mr of 54,000 . The S . coelicolor DAHP synthase was physically and kinetically characterized and the N-terminal amino acid sequence was obtained . The N-terminal amino acid sequence could not be obtained for the enzymes from S . rimosus and N . crassa, their N-termini apparently being blocked . However, following proteolytic digestion, internal amino acid sequences were obtained from both enzymes . A comparison with the known DAHP synthase sequences indicated that these DAHP synthases are unrelated to other microbial DAHP synthase sequences but are similar to plant DAHP synthases . Up until now, two distinct classes of DAHP synthase have been described, one comprising exclusively enzymes from plants, the other restricted to enzymes from micro-organisms . These studies indicate that the class containing the plant DAHP synthases also contains enzymes from a microbial eukaryote and from several bacteria. J Exp Med, 1996 Aug 1, 184(2), 747 - 52 Ligation of CD40 on dendritic cells triggers production of high levels of interleukin-12 and enhances T cell stimulatory capacity: T-T help via APC activation; Cella M et al.; We investigated the possibility that T helper cells might enhance the stimulatory function of dendritic cells (DCs) . We found that ligation of CD40 by CD40L triggers the production of extremely high levels of bioactive IL-12 . Other stimuli such as microbial agents, TNF-alpha or LPS are much less effective or not at all . In addition, CD40L is the most potent stimulus in upregulating the expression of ICAM-1, CD80, and CD86 molecules on DCs . These effects of CD40 ligation result in an increased capacity of DCs to trigger proliferative responses and IFN-gamma production by T cells . These findings reveal a new role for CD40-CD40L interaction in regulating DC function and are relevant to design therapeutic strategies using cultured DCs. J Bacteriol, 1996 Aug, 178(16), 4877 - 84 Identification and characterization of iron-regulated Bordetella pertussis alcaligin siderophore biosynthesis genes; Kang HY et al.; Bordetella bronchiseptica mutants BRM1, BRM6, and BRM9 fail to produce the native dihydroxamate siderophore alcaligin . A 4.5-kb BamHI-Smal Bordetella pertussis genomic DNA fragment carried multiple genes required to restore alcaligin production to these siderophore-deficient mutants . Phenotypic complementation analysis using subclones of the 4.5-kb genomic region demonstrated that the closely linked BRM1 and BRM9 mutations were genetically separable from the BRM6 mutation, and both insertions exerted strong polar effects on expression of the downstream gene defined by the BRM6 mutation, suggesting a polycistronic transcriptional organization of these alcaligin biosynthesis genes . Subcloning and complementation experiments localized the putative Bordetella promoter to a 0.7-kb BamHI-SphI subregion of the cloned genomic DNA fragment . Nucleotide sequencing, phenotypic analysis of mutants, and protein expression by the 4.5-kb DNA fragment in Escherichia coli suggested the presence of three alcaligin system genes, namely, alcA, alcB, and alcC . The deduced protein products of alcA, alcB, and alcC have significant primary amino acid sequence similarities with known microbial siderophore biosynthesis enzymes . Primer extension analysis mapped the transcriptional start site of the putative alcaligin biosynthesis operon containing alcABC to a promoter region overlapping a proposed Fur repressor-binding site and demonstrated iron regulation at the transcriptional level. Toxicol Lett, 1996 Aug, 86(2-3), 205 - 10 Infections and the development of allergy; Holt PG; The development of resistance or sensitivity to environmental antigens depends to a large extent on the nature of immunological memory which is generated during early antigen encounters in infancy and early childhood . The latter involves antigen-driven selection for specific Th-1-like versus Th-2-like memory cells within individual immune responses to inhaled allergens, a process which occurs in the regional lymph nodes (RLN) draining the conducting airways and which is regulated by a variety of cytokines produced by antigen-specific CD4+ and CD8+ T-cells . In addition, this T-cell selection process can theoretically be influenced by infectious agents, at two distinct levels . Firstly, infections in the airway mucosa may mobilise and activate local tissue macrophages, which migrate to RLN and secrete Th-2-inhibitory cytokines such as IL-12 and IFN alpha, and also add to IFN gamma levels in the milieu via stimulation of NK cells . Secondly, microbial stimulation via gastrointestinal tract (GIT) commensals and pathogens is recognised as the principal trigger for postnatal maturation of overall immune competence in mammals . Recent studies indicate that the speed with which the immune system in human infants attains adult-equivalent competence postnatally is inversely related to 'risk' for primary allergic sensitisation to environmental antigens . Factors which affect qualitative/quantitative aspects of microbial colonisation of the GIT during early postnatal life may accordingly have unexpected downstream effects on seemingly unrelated processes such as development of T cell memory to allergens. Science, 1996 Jul 19, 273(5273), 349 - 52 Cytoplasmic tail-dependent localization of CD1b antigen-presenting molecules to MIICs; Sugita M et al.; CD1 proteins have been implicated as antigen-presenting molecules for T cell-mediated immune responses, but their intracellular localization and trafficking remain uncharacterized . CD1b, a member of this family that presents microbial lipid antigens of exogenous origin, was found to localize to endocytic compartments that included the same specialized subset of endosomes in which major histocompatibility complex (MHC) class II molecules are proposed to bind endocytosed antigens . Unlike MHC class II molecules, which traffic to antigen-loading endosomal compartments {MHC class II compartments (MIICs)} primarily as a consequence of their association with the invariant chain, localization of CD1b to these compartments was dependent on a tyrosine-based motif in its own cytoplasmic tail. J Biotechnol, 1996 Jul 18, 48(1-2), 73 - 80 Microbial cyanide sensor for monitoring river water; Ikebukuro K et al.; A microbial cyanide sensor using Saccharomyces cerevisiae for monitoring a river water is described . This sensor is based on the inhibition of S . cerevisiae's respiration by cyanide . This sensor is a reactor type flow system and composed of two oxygen electrodes and a reactor which contains S . cerevisiae immobilized beads . The S . cerevisiae's respiration activity is measured using the oxygen electrodes . The sensor showed a linear response in the range from 0 to 15 microM and maintained stable response for 9 days at ambient temperature . The sensor was optimized for the monitoring of river water and was applied to river water analysis. Experientia, 1996 Jul 15, 52(7), 716 - 22 Two classes of metabolites from Theonella swinhoei are localized in distinct populations of bacterial symbionts; Bewley CA et al.; The marine sponge Theonella swinhoei (lithistid Family Theonellidae, Order Astrophorida) has yielded many important, bioactive natural products, most of which share structural features with bacterial natural products . The presence of microbial symbionts in T . swinhoei has been reported, and it was originally suggested that the cytotoxic macrolide swinholide A and many of the bioactive cyclic peptides from T . swinhoei were all produced by simbiotic cyanobacteria . By transmission electron microscopy, we found four distinct cell populations to be consistently present in T . swinhoei: eukaryotic sponge cells, unicellular heterotrophic bacteria, unicellular cyanobacteria and filamentous heterotrophic bacteria . Purification and chemical analyses of each cell type showed the macrolide swinholide A to be limited to the mixed population of unicellular heterotrophic bacteria, and an anti-fungal cyclic peptide occurred only in the filamentous heterotrophic bacteria . Contrary to prior speculation, no major metabolites were located in the cyanobacteria or sponge cells. FEBS Lett, 1996 Jul 8, 389(3), 263 - 7 Structural and mechanistic similarities of 6-phosphogluconate and 3-hydroxyisobutyrate dehydrogenases reveal a new enzyme family, the 3-hydroxyacid dehydrogenases; Hawes JW et al.; Rat 3-hydroxyisobutyrate dehydrogenase exhibits significant amino acid sequence homology with 6-phosphogluconate dehydrogenase, D-phenylserine dehydrogenase from Pseudomonas syringae, and a number of hypothetical proteins encoded by genes of microbial origin . Key residues previously proposed to have roles in substrate binding and catalysis in sheep 6-phosphogluconate dehydrogenase are highly conserved in this entire family of enzymes . Site-directed mutagenesis, chemical modification, and substrate specificity studies were used to compare possible mechanistic similarities of 3-hydroxyisobutyrate dehydrogenase with 6-phosphogluconate dehydrogenase . The data suggest that 3-hydroxyisobutyrate and 6-phosphogluconate dehydrogenases may comprise, in part, a previously unrecognized family of 3-hydroxyacid dehydrogenases. Compend Contin Educ Dent, 1996 Jul, 17(7), 691 - 4, 696-8 passim; quiz 709 Microbial contamination of dental unit waterlines: current preventive measures and emerging options; Williams JF et al.; Various approaches to prevention and control of microbial contamination of dental unit water are evolving . Currently available methods and emerging technologies are reviewed here in the context of contemporary federal regulatory and advisory agency positions on water quality in the dental operatory . Clean water systems, fully autoclavable systems, and a variety of devices designed to provide physical barriers to the influx and accumulation of microbial contaminants can all be used to assure satisfactory quality for coolant and irrigant water . There is one important proviso: all require a commitment to maintenance if they are to be deployed successfully . Stricter adherence to American Dental Association/American National Standards Institute standards in manufacturing and production and compliance with federal regulatory marketing procedures for equipment and devices would do much to advance the cause of dental unit water quality control. Biotechnol Prog, 1996 Jul-Aug, 12(4), 417 - 22 Problems of translating heterologous genes in expression systems: the role of tRNA; Smith DW; tRNA can have large effects on the expression and overexpression of heterologous genes in microbial expression systems through reduced translation and errors in amino acid sequences of protein products . Examples are given of large effects on gene expression related to tRNA content and to tRNA base modifications, both of which differ in heterologous expression systems compared to the cells from which the genes originally came . tRNA should be of greater concern in the expression of heterologous genes. Poult Sci, 1996 Jul, 75(7), 891 - 9 Effect of dietary fat, fiber, and monensin on cecal activity in turkeys; Zubair AK et al.; Two experiments were conducted to determine the physical, chemical, and microbial properties of turkey cecal droppings and relate them to intake of common dietary components, namely fat, fiber, and the anticoccidial, monensin . Experiment 1 involved collection and analysis of physical and chemical properties of cecal and regular droppings from commercial turkey flocks . Experiment 2 tested the effect of dietary fat, fiber, and monensin on growth performance and cecal activity in male turkeys . Compared to regular excreta, cecal droppings analyzed in Experiment 1 were higher in viscosity and fat content, and lower in dry matter, nitrogen, and fiber content (P < 0.05) . High dietary fiber and fat significantly (P < 0.05) improved growth performance in Experiment 2 . Prolonged feeding of monensin significantly (P < 0.05) reduced cecal evacuation . Results of chemical composition showed that higher dietary fat and fiber significantly (P < 0.01) reduced the fat percentage of cecal contents, whereas prolonged feeding of monensin increased the fat in both cecal contents and droppings . There was no significant effect of any of the treatments on pH, viscosity, and microbial counts of cecal contents . There is the need to identify and characterize the compound responsible for the high viscosity of cecal droppings. Patol Fiziol Eksp Ter, 1996 Jul-Sep, (3), 22 - 4 {Iron in oral cavity fluid in gingival inflammation}; Petrovich IuA et al.; In parodontosis, iron levels are higher in the human gingival fluid and mixed saliva . The free iron-binding capacity of saliva is also increased . In gingivitis, higher levels of iron and free binding capacity are less evident . The increased levels of iron activate free radical oxidation in gingival tissue and improve microbial metabolism . This all contributes to the deterioration of a pathological process in the gingiva. Diagn Microbiol Infect Dis, 1996 Jul, 25(3), 113 - 5 Prolonged incubation of blood and bone marrow cultures in 12B bottles processed on the BACTEC 460 TB system does not increase microbial recovery; Wilson ML et al.; Standard references continue to recommend testing specimens processed on the BACTEC TB System for 8 weeks, despite evidence that mycobacteria are rarely recovered beyond 5-6 weeks . To clarify this issue, we retrospectively reviewed all positive blood/bone marrow cultures processed during a 17-month period when specimens were tested for 6 weeks . We then prolonged the incubation period to 8 weeks during the subsequent 5 months . Excluding Mycobacterium genavense, only 1 of 159 mycobacterial isolates was recovered during and none were recovered beyond the 5th week of incubation and testing on the BACTEC TB System. FEMS Immunol Med Microbiol, 1996 Jul, 14(4), 221 - 30 DNA immunization: effects of vehicle and route of administration on the induction of protective antiviral immunity; Yokoyama M et al.; The effectiveness of DNA immunization has been demonstrated in several model systems, usually following intramuscular injection of DNA in saline, or topical administration to the skin . In this study we have compared DNA delivered by three routes (intramuscular, intravenous, and intraperitoneal) and, for each route, in two vehicles (cationic liposome complex and pH sensitive liposome) . These two lipid vehicles were evaluated because they are frequently used in gene therapy studies, but their immunogenicity has not been extensively studied . Each of these six combinations has been evaluated not only by assay of marker gene expression in a variety of tissues, but also by measurement of biologically-relevant parameters of immunity induction of antibodies, cytotoxic T lymphocytes, and protection against viral challenge . By both criteria (marker gene expression and induced immunity), the outcomes vary markedly among the six combinations . The combination leading to maximal marker gene expression (DNA with cationic lipid, administered i.v.) also induces detectable antibodies and CTL, and is the only one of the six combinations to induce immune responses comparable to those seen following i.m . injection of DNA in saline . However, marker gene expression can be detected in other combinations in the absence of induced immunity thus the value of marker gene expression in predicting the protection induced by a microbial antigen is questionable suggesting that, when evaluating various promoter constructs, marker gene expression may not adequately replace the direct measurement of biological outcomes. J Nihon Univ Sch Dent, 1996 Jul, 38(2), 94 - 101 Immunohistochemical characterization of lymphocyte subsets in chronic adult periodontitis; Gurses N et al.; It is well known that interactions between microbial dental plaque and the host immune system play a major role in the etiopathogenesis of periodontal disease . The aim of the present study was to analyze the phenotypic properties of gingival T lymphocytes and subsets in patients with chronic inflammatory adult periodontitis (AP) showing various degrees of inflammation and to relate the results to the immunopathogenesis of AP . Gingival biopsies were obtained from patients aged between 26 and 52 yr who were grouped according to gingival index scores (GI) of 1, 2, and 3 . Using immunohistochemical techniques, T cells (CD2+), T-helper cells (CD4+) and T-suppressor cells (CD8+) were identified in three well-defined areas of the biopsy samples . Moreover, peripheral blood was collected from the same patients, and relative counts of B cells (CD19+), HLA-DR+ cells and IL-2R+ cells as well as CD3+, CD4+, CD8+ cells were determined using three color flow cytometry . While the blood results were found to be within the normal ranges, the relative counts of CD4+ cells showed statistically significant decreases as the GI score increased . Similarly, the CD4+/CD8+ ratio also decreased . Moreover, gingival T lymphocyte and subset counts appeared to be related to the severity of gingival inflammation . Particularly, CD4+ cells showed a significant increase with the GI score . Furthermore, the CD4+/CD8+ ratio beneath the pocket epithelium was apparently correlated with increasing GI score (p < 0.05) . The cytotoxic effect of CD8+ cells seems to be more prominent at the local level while the suppressor effect is more active systematically . This means that the price of systemic protection appears to be local destruction. Br Poult Sci, 1996 Jul, 37(3), 689 - 93 Microbial spoilage of ostrich (Struthio camelus) eggs; Deeming DC; 1 . The incidence of microbial spoilage was investigated in three batches of ostrich eggs, one from Africa and two from Europe . 2 . 18% to 21% of eggs in each batch were contaminated with bacteria and fungi, with the latter being found more often in eggs which contained more advanced embryos . 3 . Although infertility and prolonged storage reduced hatchability, the high incidence of microbial contamination in ostrich eggs is deemed to be a significant problem . 4 . Improving nest hygiene is considered to be the simplest way to reduce microbial spoilage. New Microbiol, 1996 Jul, 19(3), 243 - 9 Microbial respiratory activity in the euphotic zone of the Mediterranean Sea; La Ferla R et al.; Microplankton respiration in the euphotic zone in the Central Mediterranean Sea was calculated by ETS activity measurements . Distribution of ETSa in the studied area appeared homogeneous and comparable with previous data measured in the Mediterranean Sea . A typical ETSa value for euphotic Mediterranean waters of 0.14 meq O2 h-1 m-3 was calculated . The investigated area supports a metabolic CO2 production of 70 g C m-2 y-1 . When compared with regional productivity, the respiration budget in the euphotic zone accounted for 22% of carbon fixed . ETSa appeared a useful assay for the study of evolutive history of the water masses in the Mediterranean Sea. Steroids, 1996 Jul, 61(7), 407 - 10 Microbial hydroxylation of 13-ethyl-17 beta-hydroxy-18,19-dinor-17 alpha-pregn-4-en-20-yn-3-one; Hu SH et al.; The microbial transformation of the racemic mixture of 13-ethyl-17 beta-hydroxy-18,19-dinor-17 alpha-pregn-4-en-20-yn-3-one (1) was investigated . Rhizopus nigricans (AS 3.2050), R . arrhizus (AS 3.4523), Aspergillus niger (AS 3.2744), A . ochraceus (AS 3.1408), and Curvularia lunata (NRRL 4381) transformed 1 into its 10 beta-hydroxy derivative (2) as a major metabolite . Biotransformation of 1 by Aspergillus ochraceus AS 3.1408 afforded 7 beta-hydroxy derivative (3) as the only product. J Periodontol, 1996 Jul, 67(7), 694 - 702 The dynamics of microbial colonization of barrier membranes for guided tissue regeneration; Nowzari H et al.; The microbial colonization of expanded polytetrafluoroethylene membrane by putative periodontopathogens at 3 minutes of intraoral manipulation was determined in 42 patients with 42 mandibular posterior two- to three-wall defects . Twenty patients exhibited no periodontal pockets of > or = 5 mm, other than the study site, and low levels of pathogens (group A) . Twenty-two patients revealed multiple periodontal pockets of 5 mm or more and numerous pathogens (group B) . Within the preceding 3 months of regenerative surgery, group A patients had received apically positioned flap surgery with osseous recontouring (except for the study site), and group B patients had been enrolled in a non-surgical maintenance program . The subgingival microbiota was examined prior to regenerative therapy, and the membrane microbiota was examined at 3 minutes and at the time of removal at 6 weeks by culture, DNA probes, and phase-contrast microscopy . The mean initial defect depth was 7.4 mm for group A and 7.2 mm for group B . At 6 months, the difference in mean clinical attachment gain was statistically significant (P < 0.001; group A: 3.4 mm; group B: 1.4 mm) . At 3 minutes, putative pathogens were detected in seven (16.7%) membranes in group B (group Binfected), and the associated sites gained only 0.6 mm in clinical attachment at 6 months . Clinical attachment gain was modeled as a linear function of the explanatory variables (r2 = 86%) . The presence of Porphyromonas gingivalis detected by DNA probe at 3 minutes was associated with 1.5 mm less expected gain (P = 0.0002) . Total microbial counts and the percentage of Peptostreptococcus micros and Capnocytophaga species at baseline, and of motile rods on the membrane surface facing the gingiva at 6 weeks, were statistically significant negative predictors of clinical attachment . For each week the membrane remained covered, an additional 0.5 mm gain could be expected (P = 0.002); and for every 10 sites that exhibited bleeding on probing, the clinical attachment gain was 0.6 mm less at the site of regeneration (P < 0.0001) . The present results showed that putative pathogens may colonize membranes within 3 minutes of intraoral manipulation . The patient group treated with periodontal osseous surgery revealed the lowest levels of periodontal pathogens in the membranes and exhibited the most gain in clinical attachment. J Anim Sci, 1996 Jul, 74(7), 1462 - 9 Influence of dietary magnesium level on metabolic and growth-performance responses of feedlot cattle to laidlomycin propionate; Zinn RA et al.; A metabolism trial and a growth-performance trial were conducted to evaluate the interaction of dietary magnesium level (.18 vs .32%) and laidlomycin propionate (LP) (0 vs 11 ppm, airdry basis) on utilization of a high-energy finishing diet by feedlot steers . There were no treatment effects (P > .10) on ruminal and total tract digestion of OM and ADF . However, there was an interaction (P < .05) between magnesium level and LP on ruminal starch digestion . With LP, magnesium level had no effect on ruminal starch digestion . Without LP, increasing dietary magnesium decreased ruminal starch digestion . Laidlomycin propionate decreased (P < .10) ruminal degradation of feed N (13.9%) and microbial efficiency (8.9%) and increased (P < .10) total tract N digestibility . There were no treatment interactions (P > .10) on site and extent of magnesium digestion . Magnesium absorption decreased with increased dietary magnesium (11.6%, P < .05) and LP (16.9%, P < .01) . There was an interaction (P < .10) of treatments on postabomasal calcium absorption . With the low magnesium level LP increased calcium absorption, whereas with the higher magnesium level LP decreased calcium absorption . There was an interaction between magnesium level and LP on ruminal pH at .5 h (P < .05) and 8 h (P < .10) after feeding . In general, ruminal pH increased with LP supplementation at the lower magnesium level and decreased with LP supplementation at the higher magnesium level . There were interactions (P < .05) between magnesium level and LP on ruminal VFA molar proportions and estimated methane production . At the lower magnesium level, the effects of LP on VFA molar proportions were small . At the higher magnesium level, however, LP decreased (13.2%) molar proportions of acetate and increased (26.5%) molar proportions of propionate . There was an interaction (P < .05) between magnesium level and LP on feed intake . At the lower magnesium level, LP increased (3.9%) feed intake; at the higher magnesium level LP decreased (4.4%) intake . Increasing dietary magnesium enhanced ADG (6%, P < .10) . Both magnesium and LP enhanced (2.8%, P < .10) diet NE, and this effect was strictly additive . There were no effects (P > .10) of magnesium level or LP on plasma magnesium concentrations . Plasma calcium concentrations were decreased by supplemental magnesium (15.2%, P < .05) and increased by LP (16.0%, P < .01) . We conclude that dietary magnesium levels modulate the metabolic and performance responses of feedlot steers to supplemental LP. PDA J Pharm Sci Technol, 1996 Jul-Aug, 50(4), 246 - 51 Suitability of polyvinylidene fluoride (PVDF) piping in pharmaceutical ultrapure water applications; Burkhart M et al.; This paper discusses polyvinylidene fluoride (PVDF) as an alternative to stainless steel in pharmaceutical ultrapure water piping systems . Two leading Swiss companies, a piping systems supplier and a pharmaceutical manufacturer, teamed together to jointly carryout the research . Issues addressed are metallic contamination, microbial concerns, piping system functionally and ozone resistance . Comparisons between PVDF and stainless steel are made when necessary. PDA J Pharm Sci Technol, 1996 Jul-Aug, 50(4), 201 - 4 In support of settle plates; Whyte W; During pharmaceutical production the quality of the air in manufacturing areas is monitored by volumetric samplers and settle plates . Settle plates are often assigned a secondary role, their usefulness in predicting airborne contamination of containers not being fully recognised . In this paper criticisms of settle plates are answered and the advantages of settle plates explained . It is concluded that using settle plates is the best method of quantifying the likely airborne microbial contamination of containers filled in pharmaceutical production areas. Poult Sci, 1996 Jul, 75(7), 852 - 3 Water quality and poultry production; King AJ; Mineral and microbial content of water affects the performance of poultry . Because poultry production can adversely affect water quality, the Environmental Protection Agency monitors and regulates its impact . Management of nonpoint source water contamination is especially important . If properly managed, litter, a valuable secondary commodity associated with poultry production, can be used as fertilizer, food, or energy. Physiol Behav, 1996 Jul, 60(1), 243 - 8 Salivary IgA secretion rate in young and elderly persons; Miletic ID et al.; Immunoglobulin A (IgA) is the dominant immunoglobulin isotype on all mucosal surfaces where it acts as a first line of defense against microbial invasion . Recent investigations suggest that secretory IgA (sIgA) concentrations vary over the day due to a range of variables including dietary factors, daily mood, and exercise . In this study, salivary IgA was determined by ELISA in samples of 48 persons grouped as "young" (20-30 years old) and "elderly" (60-80 years old) . Unstimulated, stabilized morning and afternoon saliva was collected during 7 consecutive days . Saliva flow, total proteins, sIgA concentrations, and sIgA secretion rates were determined . The main finding was that saliva flow and sIgA secretion rate were significantly lower in the elderly than in the young . Salivary IgA secretion rate was found to be independent of total proteins secreted in all samples . There is individual variability within a particular age group . It was also found that stress and daily events influence the sIgA secretion rate. J Antibiot (Tokyo), 1996 Jul, 49(7), 669 - 75 New betaclamycin and aclarubicin analogs obtained by prolonged microbial conversion with an aclarubicin-negative mutant; Johdo O et al.; Microbial conversion of beta-rhodomycinone and aklavinone using an aclarubicin-negative Streptomyces galilaeus mutant afforded new anthracycline antibiotics CG21-C and CG1-C which had a rednosyl-2-deoxyfucosyl-rhodosaminyl trisaccharide residue at C-7 of each added aglycone . They were produced only when a prolonged conversion culture took place . Because the usual conversion products containing a cinerulosyl-2-deoxyfucosyl-rhodosaminyl residue were at first accumulated and then decreased during further cultivation, it was evident that they occurred by the modification of terminal cinerulose . The isolation, purification, and structural determination are described, and cytotoxicity in vitro against cultured L1210 cells and the formation mechanism are discussed. Appl Environ Microbiol, 1996 Jul, 62(7), 2676 - 80 Phylogenetic compositions of bacterioplankton from two California estuaries compared by denaturing gradient gel electrophoresis of 16S rDNA fragments; Murray AE et al.; The phylogenetic compositions of bacterioplankton assemblages from San Francisco Bay and Tomales Bay, Calif., differed substantially when analyzed by PCR-denaturing gradient gel electrophoresis; these differences are consistent with the results of previous studies demonstrating differences in their metabolic capabilities . PCR-denaturing gradient gel electrophoresis analysis of complex microbial assemblages was sensitive and reliable, and the results were reproducible as shown by experiments with constructed and naturally occurring assemblages. Appl Environ Microbiol, 1996 Jul, 62(7), 2501 - 7 A computer-simulated restriction fragment length polymorphism analysis of bacterial small-subunit rRNA genes: efficacy of selected tetrameric restriction enzymes for studies of microbial diversity in nature; Moyer CL et al.; An assessment of 10 tetrameric restriction enzymes (TREs) was conducted by using a computer-simulated restriction fragment length polymorphism (RFLP) analysis for over 100 proximally and distally related bacterial small-subunit (SSU) rRNA gene sequences . Screening SSU rDNA clone libraries with TREs has become an effective strategy because of logistic simplicity, commercial availability, and economy . However, the rationale for selecting the type and number of TREs has not been systematically evaluated . Our objective was to identify the optimal combination of TREs for RFLP screening of cloned SSU rRNA genes from undefined bacterial clone libraries . After computer-simulated TRE digestion, the resultant fragments were categorized on the basis of the frequency of different restriction fragment size classes . Three groups of distribution patterns for the TREs were determined and further examined via graphical exploratory data analysis . The RFLP size-frequency distribution data for each group of enzymes were then used to infer phylogenetic relationships via the neighbor-joining method . The resulting bootstrap values and the correct placement of node bifurcations were used as additional criteria to evaluate the efficacy of the selected TREs . These RFLP data were compared with known phylogenetic relationships based on SSU rRNA sequence analysis as defined by the Ribosomal Database Project . A heuristic approach testing random combinations of TREs showed that three or more TRE combinations detected > 99% of the operational taxonomic units (OTUs) within the model data set . OTUs that remained undetected after three TRE treatments had a median sequence similarity of 96.1% . Of the 10 restriction enzymes examined, HhaI, RsaI, and BstUI (group 3) were the most efficacious at detecting and differentiating bacterial SSU rRNA genes on the basis of their ability to correctly classify OTUs . Group 3 TREs are therefore recommended for screening in studies using bacterial SSU rRNA genes as descriptors of in situ microbial diversity. FEMS Microbiol Rev, 1996 Jul, 18(4), 289 - 99 Entry of pathogens into the central nervous system; Tuomanen E; The blood-brain barrier (BBB) is formed by the tight junctions of the cerebral capillary endothelium and the choroid plexus epithelium . The molecular anatomy of the tight junction resembles that of a polarized, transporting epithelium, suggesting some model cell culture systems can provide insight into traffic into the central nervous system . Pathogens target both the endothelium, causing encephalitis, and the choroid plexus, leading to meningitis . Routes of entry are diverse including paracellular and transcellular penetration . In addition, circulating microbial products can induce loss of BBB function . Understanding the heterogeneous molecular interactions between pathogens and the BBB may provide avenues to interrupt the devastating neurological sequelae that accompany central nervous system infections. J Clin Invest, 1996 Jul 1, 98(1), 50 - 61 Soluble beta-glucan polysaccharide binding to the lectin site of neutrophil or natural killer cell complement receptor type 3 (CD11b/CD18) generates a primed state of the receptor capable of mediating cytotoxicity of iC3b-opsonized target cells; Vetvicka V et al.; When phagocyte CR3 binds to iC3b on bacteria or yeast, phagocytosis and degranulation are triggered because of simultaneous recognition of iC3b via a CD11b I-domain binding site and specific microbial polysaccharides via a lectin site located COOH-terminal to the I-domain . By contrast, when phagocyte or natural killer (NK) cell CR3 adheres to iC3b on erythrocytes or tumor cells that lack CR3-binding membrane polysaccharides, neither lysis nor cytotoxicity are stimulated . This investigation showed that soluble CR3-specific polysaccharides such as beta-glucan induced a primed state of CR3 that could trigger killing of iC3b-target cells that were otherwise resistant to cytotoxicity . Anti-CR3 added before sugars prevented priming, whereas anti-CR3 added after sugars blocked primed CR3 attachment to iC3b-targets . Polysaccharide priming required tyrosine kinase(s) and a magnesium-dependent conformational change of the I-domain that exposed the CBRM1/5 activation epitope . Unlike LPS or cytokines, polysaccharides did not up-regulate neutrophil CR3 expression nor expose the mAb 24 reporter epitope representing the high affinity ICAM-1-binding state . The current data apparently explain the mechanism of tumoricidal beta-glucans used for immunotherapy . These polysaccharides function through binding to phagocyte or NK cell CR3, priming the receptor for cytotoxicity of neoplastic tissues that are frequently targeted with iC3b and sparing normal tissues that lack iC3b. Crit Care Med, 1996 Jul, 24(7), 1157 - 62 Microbial contamination of blood conservation devices during routine use in the critical care setting: results of a prospective, randomized trial; Peruzzi WT et al.; OBJECTIVES: To compare microbial contamination of two different blood conservation devices; to determine if there was an association between contamination of the blood conservation devices and clinical infections; to determine if there was a significant user preference for either of the two devices . DESIGN: Prospective, randomized trial . SETTING: Medical, neurosurgical, and spinal cord intensive care units of an urban, university hospital . PATIENTS: Forty patients who required clinically indicated intrafierial catheters placed at new sites . INTERVENTIONS: The two most widely available blood conservation devices at the time of the study (Venous Arterial blood Management Protection system {VAMP}, Baxter Edwards Critical-Care, Irvine, CA; and Safe Draw, Ohmeda, Madison, WI) were chosen for comparison . After the normal 48 to 72 hrs of device use, the blood conservation systems were removed and semi-quantitative and quantitative cultures were taken from comparable sites of the two devices . Positive cultures from the patients were recorded and correlated with cultures obtained from the devices . In order to assess preference for either device, a survey tool was administered to the nursing staff who participated in the study . MEASUREMENTS AND MAIN RESULTS: Quantitative cultures from all sites cultured in both groups demonstrated mean colony counts of < 10(3) colony-forming units (cfu)/mL . There were no statistically significant differences in the colony counts at any of the sites compared between the two groups . There were no statistically significant relationships between positive cultures and patient age, gender, duration of device utilization, frequency of device entry, or the intensive care unit in which the study was conducted . In no circumstance did positive cultures from any of the blood conservation devices correlate with positive culture results from any sites of clinical infection . The clinical survey demonstrated a statistically significant preference for the VAMP system, which persisted despite increased experience with the Safe Draw system . CONCLUSIONS: The levels of microbial contamination noted in these devices were not consistent with clinical infection (defined as 10(3) cfu/mL on quantitative cultures) . There was no significant difference in degree or pattern of contamination between the two devices . When utilized and changed according to the Centers for Disease Control guidelines, blood conservation devices are not harbors of infection in the critical care setting . Blood conservation devices can be used as part of a comprehensive blood conservation program in the critical care setting without undue concern for exacerbating infectious processes. Microb Ecol, 1996 Jul, 32(1), 35 - 46 A Comparison of the Survival of Intraperiplasmic and Attack Phase Bdellovibrios with Reduced Oxygen Schoeffield AJ, Williams HN, Turng B, Fackler WA Jr. The ability of intraperiplasmic and attack phase bdellovibrios to survive and/or grow under anoxic and microaerobic conditions was examined . Both halotolerant and nonhalotolerant bdellovibrio strains were examined . In all instances, the bdellovibrio strains were unable to grow under anoxic conditions, but were able to survive for periods of time in both the extracellular and intraperiplasmic forms . However, the intraperiplasmic organisms were observed to survive longer . Increased temperature hastened the loss of viability of both forms of the predatory bacteria in oxic and anoxic environments . Under microaerobic conditions, halotolerant bdellovibrios were observed to grow, although at a slightly reduced rate than in atmospheric oxygen, while two nonhalotolerant isolates survived but did not grow . The ability of attack phase bdellovibrios to survive in an anoxic environment for up to nine days and their growth or survival under microaerobic conditions greatly expands the possible ecological niches in which the predators may be active members of the microbial community. Lancet, 1996 Jun 29, 347(9018), 1797 - 801 Lethal infection by a previously unrecognised metazoan parasite; Santamaria-Fries M et al.; BACKGROUND: New microbial pathogens or variant clinical manifestations of known organisms may be first found in immunodeficient patients . An HIV-infected man developed a rapidly-enlarging abdominal mass, suggestive of a neoplasm, that subsequently invaded his liver and caused death . Initial studies showed unusual tissue morphology that could not be matched with any known disease process . METHODS: Tissues obtained from biopsy at laparotomy and necropsy were studied by light microscopy, immunohistochemistry, electron microscopy, and broad-range ribosomal DNA-amplification and sequence analysis . FINDINGS: Tissue lesions were characterised by peculiar cytoplasmic sacs containing minute cells with very prominent nucleoli . The pathological process was recognised as a parasitic infection, although its features were different from those of any known eukaryotic pathogen . Phylogenetic analysis of a 357 bp 18S rDNA sequence amplified directly from the involved tissue indicated that the causative agent was a previously-uncharacterised cestode . INTERPRETATION: Fatal disease produced by this newly recognised cestode may not be limited to immunodeficient hosts . Awareness of this metazoan infection may allow early diagnosis-by morphology and DNA sequence analysis--and perhaps successful treatment of subsequent cases. Science, 1996 Jun 28, 272(5270), 1953 - 5 A Chemoautotrophically Based Cave Ecosystem Sarbu SM, Kane TC, Kinkle BK. Microbial mats discovered in a ground-water ecosystem in southern Romania contain chemoautotrophic bacteria that fix inorganic carbon, using hydrogen sulfide as an energy source . Analysis of stable carbon and nitrogen isotopes showed that this chemoautotrophic production is the food base for 48 species of cave-adapted terrestrial and aquatic invertebrates, 33 of which are endemic to this ecosystem . This is the only cave ecosystem known to be supported by in situ autotrophic production, and it contains the only terrestrial community known to be chemoautotrophically based. Eur J Biochem, 1996 Jun 15, 238(3), 822 - 9 Molecular characterization of a saline-soluble lectin from a parasitic fungus . Extensive sequence similarities between fungal lectins; Rosen S et al.; It has been proposed that the interactions between several parasitic and pathogenic fungi and their hosts are mediated by soluble lectins present in the fungus . We have cloned and analyzed a gene encoding such a lectin (AOL) from the nematophagous fungus Arthrobotrys oligospora (deuteromycete) . The deduced primary structure of the AOL gene displayed an extensive similarity (identity 46.3%) to that of a gene encoding a lectin (ABL) recently isolated from the mushroom Agaricus bisporus (basidiomycete), but not to any other fungal, microbial, plant or animal lectins . The similarities between AOL and ABL were further demonstrated by the observation that an antibody specific for AOL cross-reacted with ABL . Together with data showing that AOL has a binding specificity that is similar to that of ABL {Rosen, S., Bergstrom, J., Karlsson, K.-A . & Tunlid, A . (1996) Eur . J . Biochem . 238, 830-837}, these results indicate that AOL and ABL are members of a novel family of saline soluble lectins present in fungi . Southern blots indicated that there is only one AOL gene in the genome encoding a subunit (monomer) of the lectin . The primary structure of AOL did not show the presence of a typical N-terminal signal sequence . Comparison of the deduced primary structure with the molecular mass of AOL as determined by electrospray mass spectrometry (16153 Da), indicated that AOL has an acetylated N-terminal but no other post-translational modifications, and that a minor isoform is formed by deamidation . Circular dichroism (CD) spectroscopy suggested that the secondary structure of AOL contains 34% beta-sheets, 21% alpha-helix, and 45% turns and coils. J Immunol, 1996 Jun 15, 156(12), 4570 - 5 Bacterial DNA induces NK cells to produce IFN-gamma in vivo and increases the toxicity of lipopolysaccharides; Cowdery JS et al.; Microbial products released during bacterial infection induce cytokine-mediated inflammatory responses that can be protective, but excessive release of inflammatory cytokines may promote development of the sepsis syndrome . We examined the ability of bacterial DNA to induce in vivo cytokine release and to potentiate the toxicity of LPS . Intravenous treatment of mice with Escherichia coli (EC) DNA, but not calf thymus (CT) DNA, induced a rapid (within 4 h) dose-dependent increase in serum IFN-gamma and splenic IFN-gamma-forming cells . Over 90% of splenic IFN-gamma-producing cells were identified by surface phenotype as NK cells . Mice also mounted an IFN-gamma response following challenge with 20-base oligonucleotide that contained an internal CG motif (but did not respond to a control oligonucleotide) . Treatment of mice with EC DNA followed by a sublethal LPS challenge resulted in a 3-fold increase in the peak serum level of TNF-alpha and a 10-fold increase in the peak level of IL-6 compared with mice that received CT DNA followed by LPS . Mice treated with EC DNA followed by LPS showed 75% mortality, compared with no deaths in mice treated with CT DNA followed by LPS . EC DNA/LPS treatment of mice with disrupted IFN-gamma genes resulted in a 5% mortality while 59% of similarly treated +/+ mice died . Thus, bacterial DNA induces in vivo release of IFN-gamma which, in turn, is associated with an increase in LPS-induced TNF-alpha and IL-6 release, and with increased sensitivity to the toxic effects of LPS. Mutat Res, 1996 Jun 12, 363(2), 97 - 104 Purification and characterization of Drosophila melanogaster photolyase; Kim ST et al.; Animal-type photolyases have very limited sequence homology to microbial-type photolyases . We wanted to find out whether the two types of enzymes have different or similar biochemical and photochemical properties . In particular, the chromophore/cofactor composition of animal photolyases is of special interest since the presence and nature of a second chromophore in these enzymes are not known in contrast to the microbial photolyases which contain FAD cofactor, and folate or deazaflavin as second chromophores . We overproduced the Drosophila melanogaster photolyase in Escherichia coli using the cloned gene . The enzyme contains FAD and folate and thus belongs in the folate class of enzymes but with an action spectrum peak at 420 nm. J Biol Chem, 1996 Jun 7, 271(23), 13724 - 30 Site-directed mutagenesis of Cys-15 and Cys-20 of pulmonary surfactant protein D . Expression of a trimeric protein with altered anti-viral properties; Brown-Augsburger P et al.; Surfactant protein D (SP-D) molecules are preferentially assembled as dodecamers consisting of trimeric subunits associated at their amino termini . The NH2-terminal sequence of each monomer contains two conserved cysteine residues, which participate in interchain disulfide bonds . In order to study the roles of these residues in SP-D assembly and function, we employed site-directed mutagenesis to substitute serine for cysteine 15 and 20 in recombinant rat SP-D (RrSP-D), and have expressed the mutant (RrSP-Dser15/20) in Chinese hamster ovary (CHO-K1) cells . The mutant, which was efficiently secreted, bound to maltosyl-agarose, but unlike RrSP-D, was assembled exclusively as trimers . The constituent monomers showed a decreased mobility on SDS-polyacrylamide gel electrophoresis resulting from an increase in the size and sialylation of the N-linked oligosaccharide at Asn-70 . Although RrSP-Dser15/20 contained a pepsin-resistant triple helical domain, it showed a decreased Tm, and acquired susceptibility to proteolytic degradation . Like RrSP-D, RrSP-Dser15/20 bound to the hemagglutinin of influenza A . However, it showed no viral aggregation and did not enhance the binding of influenza A to neutrophils (PMN), augment PMN respiratory burst, or protect PMNs from deactivation . These studies indicate that amino-terminal disulfides are required to stabilize dodecamers, and support our hypothesis that the oligomerization of trimeric subunits contributes to the anti-microbial properties of SP-D. Pharm Pract Manag Q, 1996 Jul, 16(2), 4 - 8 Cost and quality considerations in antibiotic formulary management; Ritchie DF; Numerous factors are considered in the design of an antibiotic formulary, consisting primarily of indicators of clinical efficacy and safety, as well as economic viability . Similarly, there are many potential strategies for subsequent antibiotic formulary management ranging from global educational efforts to comprehensive required antibiotic justification programs . Antibiotic formulary management programs should be tailored to individual institutions and utilize a multidisciplinary approach where possible . The primary outcome goals of such programs are generally controlled antibiotic costs, optimized clinical efficacy, minimization of adverse events, and reduced microbial resistance development, although quantification of program impact on these parameters continues to be a challenge. Minerva Chir, 1996 Jun, 51(6), 443 - 6 {Experimental study of postoperative lavage: standardization of the method}; Sortini A et al.; Postoperative infections are an outstanding problem in a surgical department . We have been studying them from a clinical and experimental point of view has a long time . In this study we present a method standardisation of postoperative peritoneal lavage as prevention of surgical infections . Winstar mice, infected with intra peritoneal E . coli cultures, underwent peritoneal lavage with saline solution or sterile water . The results showed the right amount of solution and the exact administration of lavage according to grade of infection . Encouraged by this result we are going to test the efficacy of anti microbial agents for the postoperative peritoneal lavage. Oral Dis, 1996 Jun, 2(2), 167 - 80 Osteoclast activation in inflammatory periodontal diseases; Wiebe SH et al.; OBJECTIVE: In this paper, we review the mechanisms thought to be involved in the activation of osteoclasts in periodontitis . SUMMARY: Osteoclasts are regulated by both microbial and host factors . Some factors act directly on cells of the osteoclast lineage, whereas others act indirectly through other cell types in the bone environment . The proinflammatory cytokines (interleukins 1 and 6, tumor necrosis factors) have been implicated in the stimulation of osteoclastic resorption . The roles of the immunoregulatory cytoknes (interleukins 2 and 4, interferon gamma) are less clear, but decreased levels of these factors may contribute to periodontitis . A number of lipid mediators may be involved in stimulation of bone resorption . These include bacterial lipopolysaccharide and host-derived platelet-activating factor and prostaglandins . More recently, reactive oxygen intermediates and extracellular nucleotides, both present at sites of inflammation, have been investigated as possible modulators of osteoclast activity . The potential use of antiresorptive therapies in periodontitis is reviewed . CONCLUSIONS: A wide range of host and bacterial factors contribute to the loss of alveolar bone in periodontitis . However, much remains to be understood about the complex mechanisms through which these factors regulate osteoclast activity . Further studies at the cellular and molecular level will lead to a better understanding of these processes and perhaps suggest new approaches for periodontal therapy. Oral Dis, 1996 Jun, 2(2), 129 - 34 Peroxidases, lactoferrin and lysozyme in peripheral blood neutrophils, gingival crevicular fluid and whole saliva of patients with localized juvenile periodontitis; Suomalainen K et al.; OBJECTIVE: The aim of this study was to examine the longitudinal association of selected non-immune anti-microbial host factors (peroxidases, lysozyme and lactoferrin) to the localized juvenile periodontitis (LJP) disease status . MATERIALS AND METHODS: Peroxidases, lysozyme and lactoferrin were quantitated from seven patients with LJP before and after periodontal therapy . Analyses were performed from simultaneously collected samples of peripheral blood polymorphonuclear leukocytes (PMNs), gingival crevicular fluid (GCF from diseased sites) and paraffin-stimulated whole saliva . Similar assays were done also from seven periodontally healthy controls . RESULTS: During untreated phase of LJP myeloperoxidase, lysozyme and lactoferrin concentrations were remarkably elevated in peripheral blood PMNs, also reflected in their high concentrations in GCF . All these values normalised with respect to healthy controls during the periodontal therapy . No similar longitudinal changes were seen in whole saliva but during therapy salivary peroxidase concentrations declined below the control values, in accordance with our previous observations in parotid saliva samples of LJP patients . CONCLUSIONS: In LJP the concentrations of lysozyme, lactoferrin and myeloperoxidase are significantly elevated in peripheral blood PMNs, also reflected in GCF . During periodontal therapy these values decline and approach those observed in healthy controls . No similar changes are seen in stimulated whole saliva. Cytokine Growth Factor Rev, 1996 Jun, 7(1), 47 - 64 Chemokine receptors: structure, function and role in microbial pathogenesis; Murphy PM; The chemokine superfamily is composed of at least 20 different leukocyte chemoattractants that act by binding to a family of G protein-coupled receptors . Leukocyte subtypes respond preferentially to unique but overlapping subsets of chemokines as determined by the receptor distribution, yet the receptors appear to signal through a common Gi-type G protein . Since chemokines appear to play major roles in inflammatory pathology, their receptors may be good targets for developing leukocyte selective anti-inflammatory drugs . Two chemokine receptors, CC CKRS and ONCC, function pathologically as cell entry factors respectively for human immunodeficiency virus 1, the cause of AIDS, and Plasmodium vivax, the major cause of malaria. Int J Food Microbiol, 1996 Jun, 30(1-2), 157 - 73 Incorporation of elements of quantitative risk analysis in the HACCP system; Notermans S et al.; Foodborne bacterial diseases cause considerable morbidity and mortality throughout the world . Preventive measures such as good manufacturing practices (GMP), supplemented by the hazard analysis critical control point (HACCP) system, have been introduced as a means of ensuring the production of safe food . However, their use does not necessarily provide quantitative information on the risks associated with the consumption of a particular food product . To obtain such information, elements of quantitative risk analysis (QRA) need to be used . QRA is defined as a stepwise analysis of the health risks associated with a specific type of food product, resulting in an estimation of the probability of occurrence of adverse effects on health following consumption of the food in question . It also includes an analysis of the nature of the risks . Taking this definition, five successive steps can be recognized: hazard identification, exposure assessment, dose response assessment, risk characterization and risk management . Food production is a dynamic activity, involving changes in, e.g . the composition and microbial quality of raw materials due to seasonal variation . Also, there may be continuing changes in processing conditions and in product composition due to consumer demands . Therefore, it will be desirable to incorporate QRA in existing safety assurance systems, such as HACCP, when sufficient information is available to permit this approach. J Steroid Biochem Mol Biol, 1996 Jun, 58(3), 337 - 49 Microbial transformations of steroids--X . Cytochromes P-450 11 alpha-hydroxylase and C17-C20 lyase and a 1-ene dehydrogenase transform steroids in Nectria haematococca; Ahmed F et al.; Nectria haematococca contains four enzymes that metabolise exogenous steroids . Two of these are microsomal cytochromes P-450 which act sequentially on progesterone producing firstly, by side-chain cleavage, the C19 steroid androstenedione (C17-C20 lyase), and then, in a subsequent set of transformations, 11 alpha-hydroxylated derivatives (11 alpha-hydroxylase) . Two other conversions occur after side-chain cleavage . Unsaturation, in the form of a double bond at C1-C2, is introduced into the A ring by a catalytically self-sufficient microsomal 1-ene dehydrogenase . This enzyme is specific for C19 substrates . A C17-specific oxidoreductase is also involved in the production of androstenedione and testosterone from progesterone . The lyase, 11 alpha-hydroxylase and 1-ene dehydrogenase were purified to homogeneity. Drug Saf, 1996 Jun, 14(6), 386 - 93 A risk-benefit assessment of agents used in the treatment of scabies; Elgart ML; Permethrin is probably the most effective topical treatment for scabies . There is little microbial resistance to the drug, and it is highly effective . Toxicity is limited to occasional contact dermatitis . Similarly, oral ivermectin 200 micrograms/kg is extremely effective . Oral administration eliminates the need to be certain that medication has been applied properly . Toxicity has been very limited . It is not available in the US for human use in scabies at this time . The toxicity of most of other treatments that are available has not been studied carefully . Sulphur 6% in petrolatum is recommended as safe, but there are no good studies to confirm this . Sulphur is probably the medication of choice when cost is the overriding concern . Crotamiton and benzyl benzoate are probably safe, but are not as effective as permethrin . Lindane has some potential CNS toxicity if used incorrectly, and must be used carefully on damaged skin to avoid excessive absorption . It is important to stress that all household and sexual contacts must be treated, whether or not they have symptoms . Proper application of topical medications must be achieved, including under the fingernails, and up to the edge of all body orifices. Clin Otolaryngol, 1996 Jun, 21(3), 269 - 73 The detection of Mycoplasma pneumoniae in nasal polyps; Gurr PA et al.; The aetiology and microbial flora of nasal polyps is not well understood . No study in the literature has reported an association between the sub-bacterium Mycoplasma pneumoniae and nasal polyps . We have developed an assay method using the Polymerase Chain Reaction (PCR) to amplify a specific region of the M . pneumoniae DNA in extracts of clinical samples using species-specific primers designed to a region of the 16S rRNA . The presence of M . pneumoniae was detected in 13/14 (93%) nasal polyps, in 4/5 (80%) rhinosinusitis mucosal samples but only in 1/7 (14%) of control samples (obstructive turbinates) . An epidemic of infections due to M . pneumoniae is expected to occur in 1995 . We believe this assay could form the basis of a rapid technique for M . pneumoniae detection . We also propose that the presence of M . pneumoniae may be of importance in the aetiology of nasal polyps. Methods, 1996 Jun, 9(3), 563 - 77 The O-2 Generating NADPH Oxidase of Phagocytes: Structure and Methods of Detection Edwards SW. The NADPH oxidase of phagocytic cells of the immune system plays an important role in the destruction of certain types of microbial pathogens during infections . Its inappropriate activation to release reactive oxidants may also contribute to host tissue damage in inflammatory diseases . In this review, the structure of the NADPH oxidase is described and many of the commonly used methods to detect its activation during the respiratory burst are listed . The advantages and disadvantages of each of these methods are critically discussed. J Vet Med Sci, 1996 Jun, 58(6), 537 - 41 Elevation of serum G-CSF level in horses with transportation-induced fever; Momoi Y et al.; Levels of granulocyte-colony stimulating factor (G-CSF) in the blood of horses were measured before and after a long-distance transportation to clarify the pathogenesis of transportation-induced fever . The serum G-CSF level was measured by its ability to stimulate growth in a mouse myeloblastic cell line, NFS-60 . Of 26 horses transported for a long distance, 9 had fever more than 39.0 degrees C during or after transportation . After transportation, the serum G-CSF level significantly increased in horses with transportation-induced fever but not in those without fever, and the serum G-CSF level correlated positively with the peak body temperature and with an increase in peripheral white blood cell count . These data indicate that microbial infection, which is closely related to the elevation of the serum G-CSF levels, is the causative factor of transportation-induced fever. Mol Med Today, 1996 Jun, 2(6), 258 - 67 Molecular and cellular basis of interleukin 12 activity in prophylaxis and therapy against infectious diseases; Gazzinelli RT; A major goal for immunologists dealing with infectious diseases is the development of vaccines and immunotherapies that will protect patients against infection or the undesired effects of immune responses elicited by pathogens . Studies defining the function of different cytokines have contributed to the progress of new strategies to manipulate the immune response . Recent studies have demonstrated that interleukin 12 (IL-12) is a key cytokine for promoting cell-mediated immunity and initiating resistance to infection . Because IL-12 is a potent stimulator of host defense against a variety of pathogens, it holds great promise for therapeutic use . In addition, IL-12 antagonists protect the host against immunopathology and death caused by an excessive cellular immune response that can occur during acute microbial infections. Mol Med Today, 1996 Jun, 2(6), 250 - 7 Animal models of antisense oligonucleotides: lessons for use in humans; Plenat F; The ability of oligonucleotides to inhibit genetic expression in a sequence-specific manner has been well documented . Because of their potential for exquisite specificity, oligonucleotides have been proposed as therapeutic agents for a variety of human diseases, including cancers, microbial infections and autoimmune disorders . Approximately 16 clinical trials are currently in progress . However, relatively little is known about the in vivo behaviour of oligonucleotides . Extrapolations from in vitro studies to predict in vivo pharmacokinetics and effects in humans might be difficult and inappropriate . Animal models still remain an essential tool in the development of oligonucleotides as efficient drugs in humans. J Periodontol, 1996 Jun, 67(6), 577 - 82 Efficacy of a 2-phase oil: water mouthrinse in controlling oral malodor, gingivitis, and plaque; Kozlovsky A et al.; The purpose of the study was to examine the anti-malodor, anti-gingivitis, and plaque reducing properties of a 2 phase oil:water mouthrinse compared with a control mouthrinse . Fifty subjects rinsed with one of the two rinses for 30 seconds twice a day over 6 weeks, while continuing their normal oral hygiene habits . Measurements were made at time zero (prior to beginning the rinsing regimen), and > or = 9 hours following rinsing, at intervals of 1, 3, and 6 weeks . Malodor of whole mouth, as well as tongue dorsum anterior and posterior, was assessed on a 0 to 5 semi-integer scale by two odor judges . Volatile sulphide compounds (VSC) were determined using a sulphide monitor . Gingival, plaque, and bleeding indices were recorded for Ramfjord teeth . Oral microbial levels were assessed using the oratest . Salivary levels of diamines (putrescine and cadaverine) were analyzed by HPLC . Results were analyzed by 2-tailed covariant ANOVA, with the time zero value as covariant . Dramatic improvements were observed in parameters associated with malodor, periodontal health, plaque accumulation, and microbial levels in both groups . As compared to time zero scores, whole mouth odor, tongue dorsum anterior and posterior odors decreased continuously over time, attaining 80%, 79% and 70%, reductions, respectively following 6 weeks, in the 2-phase mouthrinse group, versus 70%, 77% and 59% for the control group . For whole mouth and tongue dorsum posterior, the reductions observed in the 2-phase mouthrinse group were significantly greater than those obtained with the control mouthrinse (P = 0.026 and P = 0.025, respectively), suggesting that the 2-phase mouthrinse is superior to the control mouthrinse in long-term reduction of oral malodor . For bleeding index, gingival index, oral microbial levels, and VSC, differences between the groups were not significant . Diamine levels were not significantly reduced in either group . The control mouthrinse reduced plaque index more significantly than the 2-phase mouthrinse (P < 0.005) . The results of this randomized clinical trial suggest that the 2-phase oil:water mouthrinse formulation is superior to the control mouthrinse in long-term reduction of oral malodor. J Arthroplasty, 1996 Jun, 11(4), 469 - 73 Efficacy of the Steri-Shield filtered exhaust helmet in limiting bacterial counts in the operating room during total joint arthroplasty; Shaw JA et al.; Filtered exhaust helmets (space suits) have been reported as efficacious in decreasing wound infections following joint arthroplasty procedures; however, strict experimental control of the many variables related to the incidence of clinical infection has been lacking in these studies, making support of such conclusions difficult . Any potential benefit of filtered exhaust helmets in the control of infection rates can be logically assigned to the reduction of airborne bacterial counts within the operating room . A study was done using the Steri-Shield helmet (Bio-Medical Devices, Irvine, CA) to define its efficacy in limiting airborne bacterial contaminates during total joint arthroplasty . Forty-eight paired prosthetic hip and knee cases were evaluated using a block experimentation design . One case of each pair was done with all surgical and scrub personnel cloaked in Steri-Shield filtered exhaust helmets and the other with the same personnel in conventional paper hoods and masks . Air samples were obtained with an Anderson two-stage viable particle sampler (Anderson Samplers, Atlanta, GA) placed immediately adjacent to the wound . Quantitative microbial cultures were obtained . The filtered exhaust helmet evaluated in this study provided no increased protection against bacterial contamination in the area of the surgical field than conventional paper hoods and masks. Gastroenterologist, 1996 Jun, 4(2), 129 - 33 Pouchitis: pathogenesis, diagnosis, and management; Rubinstein MC et al.; The current treatment of choice for patients requiring colectomy for ulcerative colitis or familial adenomatous polyposis (FAP) is ileoanal anastomosis with pouch creation . Symptomatic inflammation of this pouch, a condition known as pouchitis, will develop in up to 40% of patients who undergo this surgery . Patients will present with crampy abdominal pain, fever, rectal bleeding, and diarrhea, and they may have either acute intermittent attacks or a chronic pouchitis syndrome . Most reported cases of pouchitis have occurred in patients with a previous history of ulcerative colitis, whereas complications develop in only a handful of patients with FAP . The etiology of pouchitis is probably a multifactorial event involving genetic, immune, microbial, and toxic mediators . The initial medical management of pouchitis usually relies on metronidazole; however, other drugs that are useful for ulcerative colitis have been found to be beneficial for pouchitis . Studying the etiology and management of pouchitis may help elucidate the pathogenesis of inflammatory bowel disease. Appl Environ Microbiol, 1996 Jun, 62(6), 1998 - 2005 Identification in situ and dynamics of bacteria on limnetic organic aggregates (lake snow); Weiss P et al.; Microbial assemblages on large organic aggregates (lake snow) of Lake Constance, Germany, were analyzed with rRNA-directed fluorescent oligonucleotide probes specific for the domain Bacteria and the alpha-, beta-, and gamma-subclasses of the class Proteobacteria . Lake snow aggregates were either collected in situ by SCUBA diving or in a sediment trap at 50 m or formed of natural lake water incubated in rolling cylinders under simulated in situ conditions . For the latter aggregates, the time course of the microbial colonization was also examined . The natural aggregates and those made in rolling cylinders were composed of the particulate organic material present in the lake and thus reflected the composition of the ambient plankton community . All types of lake snow aggregates examined were heavily colonized by microbial cells and harbored between 0.5 x 10(6) and > 2 x 10(6) cells aggregate -1 . Between 55 and 100% of the microbial cells stained with 4', 6-diamidino-2-phenylindole (DAPI) could be visualized with the domain Bacteria-specific probe . In most samples, beta-subclass proteobacteria dominated the microbial community, constituting 27 to 42% of total cells as counted by DAPI staining, irrespective of the composition of the aggregates . During the time course experiments with the laboratory-made aggregates, the fraction of beta-subclass proteobacteria usually increased over time . Except for a few samples, alpha- and gamma-subclass proteobacteria were far less abundant than beta-subclass proteobacteria, constituting 11 to 25 and 9 to 33% of total cells, respectively . Therefore, we assume that a specific aggregate-adapted microbial community was established on the aggregates . Because the compositions of the microbial assemblages on natural and laboratory-made aggregates were similar, we conclude that aggregates made in rolling cylinders are good model system with which to examine the formation and microbial colonization of macroscopic organic aggregates. Appl Environ Microbiol, 1996 Jun, 62(6), 1889 - 96 Characterization of a Neocallimastix patriciarum cellulase cDNA (celA) homologous to Trichoderma reesei cellobiohydrolase II; Denman S et al.; The nucleotide sequence of a cellulase cDNA (celA) from the rumen fungus Neocallimastix patriciarum and the primary structure of the protein which it encodes were characterized . The celA cDNA was 1.95 kb long and had an open reading frame of 1,284 bp, which encoded a polypeptide having 428 amino acid residues . A sequence alignment showed that cellulase A (CELA) exhibited substantial homology with family B cellulases (family 6 glycosyl hydrolases), particularly cellobiohydrolase II from the aerobic fungus Trichoderma reesei . In contrast to previously characterized N . patriciarum glycosyl hydrolases, CELA did not exhibit homology with any other rumen microbial cellulases described previously . Primary structure and function studies in which deletion analysis and a sequence comparison with other well-characterized cellulases were used revealed that CELA consisted of a cellulose-binding domain at the N terminus and a catalytic domain at the C terminus . These two domains were separated by an extremely Asn-rich linker . Deletion of the cellulose-binding domain resulted in a marked decrease in the cellulose-binding ability and activity toward crystalline cellulose . When CELA was expressed in Escherichia coli, it was located predominantly in the periplasmic space, indicating that the signal sequence of CELA was functional in E.coli . Enzymatic studies showed that CELA had an optimal pH of 5.0 and an optimal temperature of 40 degrees C . The specific activity of immunoaffinity-purified CELA against Avicel was 9.7 U/mg of protein, and CELA appeared to be a relatively active cellobiohydrolase compared with the specific activities reported for other cellobiohydrolases, such as T . reesei cellobiohydrolases I and II. Biodegradation, 1996 Jun, 7(3), 203 - 21 Transformations of six isomers of dimethylbenzothiophene by three Pseudomonas strains; Kropp KG et al.; Dimethylbenzothiophenes are among the sulfur heterocycles in petroleum that are known to be degraded by microbial activity . Six of the 15 possible isomers of dimethylbenzothiophene were synthesized and used in bio-transformation studies with three Pseudomonas isolates that oxidize a variety of condensed thiophenes including methylbenzothiophenes and methyldibenzothiophenes . The isomers of dimethylbenzothiophene were chosen to have a variety of substitution patterns: both methyl groups on the thiophene ring (the 2,3-isomer); a methyl group on each of the rings (the 2,7-, 3,5-and 3,7-isomers); and both methyl groups on the benzene ring (the 4,6-and 4,7-isomers) . Each isolate was grown on 1-methylnaphthalene or glucose in the presence of one of the dimethylbenzothiophenes and culture extracts were analyzed to identify nearly 30 sulfur-containing metabolites in total . Sulfoxides and sulfones were commonly found metabolites in culture extracts from the 2,3-, 2,7- and 3,7-isomers, whereas 2,3-diones, 3(2H)-ones and 2(3H)-ones were formed from the 4,6- and 4,7-isomers . High-molecular-weight products, some of which were tentatively identified as tetramethylbenzo{b}naphtho{1,2-d}thiophenes, were detected in the extracts of cultures incubated with 4,6- or 4,7-dimethylbenzothiophene . The methyl groups of all of the isomers, except 4,6-, were oxidized to give hydroxymethyl-methylbenzothiophenes and methylbenzothiophene-carboxylic acids, and these were the only products detected from the oxidation of 3,5-dimethylbenzothiophene. Br J Nutr, 1996 Jun, 75(6), 839 - 52 Response of broilers to graded levels of microbial phytase added to maize-soyabean-meal-based diets containing three levels of non-phytate phosphorus; Kornegay ET et al.; Male 1-d-old broilers (n 920) were given 0, 200, 400, 600, 800, 1000 and 1200 U microbial phytase/kg diet in combination with 2.0, 2.7 or 3.4 g non-phytate P (nP)/kg or 4.0, 5.1 or 5.8 g total P (tP)/kg in a 21 d trial to assess the effectiveness of phytase in a maize-soyabean-meal diet . In addition to the above twenty-one diets, a positive control P diet supplied 4.5 g nP/kg, 6.9 g tP/kg and 10 g Ca/kg . The basal diet contained 230 g crude protein/kg, 8.8 g Ca/kg, 4.4 g tP/kg and 2.0 g nP/kg . Defluorinated phosphate and limestone were used to supply P and Ca . A Ca:tP ratio of 2:1 was maintained except in the positive control diet which had a ratio of 1.45:1 . Phytase additions linearly increased (P < 0.01) body-weight (BW) gain, feed intake, toe ash percentage, and apparent retention (% of intake) or total amount (g/bird) of retained Ca and P, and linearly decreased (P < 0.01) P excretion (g/kg of DM intake) at each level of nP with the magnitude of the response inversely related to the level of nP . Above-normal mortality was only observed in the group receiving 2.0 g nP/kg diet without phytase . Adding nP linearly increased (P < 0.01) BW gain, feed intake, toe ash percentage, Ca retention, total amount (g/bird) of P retained, and P excretion, and linearly decreased (P < 0.01) apparent retention (%) of P . Derived linear and non-linear equations for BW gain and toe ash percentage at the two lower nP levels, 2.0 and 2.7 g/kg, were used to calculate P equivalency values of microbial phytase . The results show that 939 U microbial phytase is equivalent to 1 g P from defluorinated phosphate in broilers fed on maize-soyabean-meal diets . The amount of P released per 100 U phytase decreased as the total amount of phytase increased. Unfallchirurgie, 1996 Jun, 22(3), 139 - 42 {Systematic revision--a contribution to the treatment strategy of early infection in trauma surgery}; Eyssel M et al.; Infections of the soft tissue and joint are the serious complication in traumatology . We are presenting our new surgical planned revision concept based on the clinical findings and the microbial sensitivity test . This concept shows that a number of radical debridements influencing by the local findings and the sterile results of 2 following cytologic smears led us to a higher certainty in healing surgical soft tissue and joint infection. Appl Microbiol Biotechnol, 1996 Jun, 45(5), 600 - 6 Purification and some properties of a novel microbial lactate oxidase; Sztajer H et al.; Geotrichum candidum was found to produce a lactate oxidase . The enzyme was purified by gel filtration and ion-exchange chromatography . The purified lactate oxidase showed a molecular mass of 50 kDa under denaturing and about 400 kDa under non-denaturing conditions . Transmission electron microscopy analysis confirmed an octameric structure . FMN was found to be a cofactor for this enzyme . Polarographic studies confirmed an oxygen uptake by the lactate oxidase . The enzyme showed specificity towards the L isomer of lactate and did not oxidise pyruvate, fumarate, succinate, maleate and ascorbate . It was stable at alkaline pH and also for 15 min at 45 degrees C . The addition of glycerol and dextran 500,000 to the enzyme sample enhanced storage stability. Int J Biol Macromol, 1996 Jun, 18(4), 307 - 9 Some novel N-fatty acyl derivatives of a microbial galactosaminan; Zhang M et al.; Novel seven N-fatty acyl derivatives (degree of substitution 0.78-0.96) of a microbial galactosaminan were prepared in 59-79% yields by its reaction with fatty acid anhydrides in aqueous acetic acid-methanol . N-Acetyl and N-propionyl derivatives were soluble in water, aqueous 2% sodium hydroxide, and aqueous 2% acetic acid, but N-higher fatty acyl (> C6) derivatives were insoluble . Gel was not formed in these reactions. Poult Sci, 1996 Jun, 75(6), 729 - 36 The effects of supplemental microbial phytase on the performance and utilization of dietary calcium, phosphorus, copper, and zinc in broiler chickens fed corn-soybean diets; Sebastian S et al.; A 3-wk feeding trial with 180 sexed day-old broiler chickens was conducted to study the efficacy of microbial phytase (Natuphos 1000) on growth performance, relative retention of P, Ca, Cu, and Zn, and mineral contents of plasma and bone . Treatments involved a normal P level corn-soybean diet, a low-P diet, and a low-P plus phytase (600 phytase units/kg) diet . Phytase supplementation increased (P < or = 0.05) body weight in male and female chickens by 13.2 and 5.8%, respectively, at 21 d . The improvements yielded body weights comparable to those obtained on the normal P diet . Phytase supplementation overcame (P < or = 0.05) the depression of feed intake observed on the low-P diet . Treatments had no effect on feed:gain ratio . Phytase supplementation of the low-P diet increased (P < or = 0.05) the relative retention of total P, Ca, Cu, and Zn by 12.5, 12.2, 19.3, and 62.3 percentage units, respectively, in male chickens . Microbial phytase increased the plasma P by 15.7% and reduced (P < or = 0.05) the Ca concentration by 34.1%, but had no effect on plasma concentrations of Cu or Zn . Phytase supplementation increased the percentage ash in both head and shaft portions of dry, fat-free tibia bone to a level comparable to that of the normal-P diet . Phytase supplementation had no effect on the concentration of any of the minerals measured in whole tibia ash but did increase (P < or = 0.05) the DM percentage of P and Ca min tibia head of male chickens by 0.65 and 1.4 percentage units, respectively . These results show that microbial phytase supplementation of a low-P diet increased growth and relative retention of total P, Ca, Cu, and Zn and improved bone mineralization in broiler chickens. Leuk Res, 1996 Jun, 20(6), 491 - 7 Angelmicin B, a new inhibitor of oncogenic signal transduction, inhibits growth and induces myelomonocytic differentiation of human myeloid leukemia HL-60 cells; Yokoyama A et al.; Angelmicin B is a new microbial substance which inhibits src tyrosine kinase activity and oncogenic signal transduction . We investigated the effect of angelmicin B on the proliferation and differentiation of the HL-60 human myeloid leukemia cell line . Angelmicin B caused the dose-dependent inhibition of cell proliferation and induction of differentiation along the myelomonocytic pathway, as determined by morphological changes, nitroblue tetrazolium (NBT) reduction, and non-specific esterase and lysozyme activities at concentrations ranging from 0.1 to 0.5 microgram/ml . Also, it induced significantly the differentiation of mouse myeloid leukemia M1 cells . A similar concentration of angelmicin B inhibited the growth of the myeloid leukemia cell lines K562, HEL, KU812, ML-1, U937 and THP-1, but did not induce differentiation of these cells significantly . The differentiation of HL-60 cells was enhanced by combined treatment with angelmicin B and 1 alpha, 25-dihydroxyvitamin D3 (VD3), retinoic acid or tumor necrosis factor-alpha (TNF alpha) . Angelmicin analogs (A1, A2, B, C and D) had almost equivalent effects on the differentiation of HL-60 cells, although angelmicins C and D inhibited src tyrosine kinase activity less than the other analogs . The effective concentrations of angelmicin B in src kinase inactivation was about 100-fold higher than those required for the growth inhibition and differentiation induction . These findings indicate that the differentiation-inducing activity of angelmicins is not associated with their src kinase-inhibiting activity, and may be associated with the modulation of other signal pathway(s). J Appl Bacteriol, 1996 Jun, 80(6), 617 - 25 Microbial biomass in a shallow, urban aquifer contaminated with aromatic hydrocarbons: analysis by phospholipid fatty acid content and composition; Franzmann PD et al.; The city of Perth contains a number of sites that have been contaminated with hydrocarbons due to leakage from petroleum underground storage tanks . Microbial biomass in groundwater and sediment cores from above and below the water table, and from within and outside a plume of hydrocarbon contamination, was examined using phospholipid fatty acid methyl ester analysis . Microbial numbers, calculated from the phospholipid content, ranged from 0.9 x 10(6) to 7.8 x 10(6) 'Escherichia coli equivalent cells' g-1 dry wt of sediment . Over 96% of the microbial biomass was attached to the sediment and the proportion of attached cells did not decrease within the plume of contaminants . The amount of biomass within aquifer samples seemed to be related more to the proximity of the rhizosphere to the shallow aquifer, and other unknown urban inputs, rather than to the effects of the plume of contaminants . Fatty acids common to many bacterial groups dominated within the plume, and as such the analyses gave limited insight into microbial community structure . For site assessment of intrinsic remediation of shallow aquifers in urban areas, estimates of microbial biomass may not provide information that is readily applicable to plume management. J Clin Anesth, 1996 Jun, 8(4), 329 - 36 Compatibility of propofol, fentanyl, and vecuronium mixtures designed for potential use in anesthesia and patient transport; Isert PR et al.; STUDY OBJECTIVE: To determine whether propofol emulsion, fentanyl, and vecuronium remain compatible and stable when mixed in clinically appropriate concentrations . DESIGN: Seven separate in vitro experiments . Two different propofol-fentanyl-vecuronium mixtures were formulated . Regular assays of propofol, fentanyl, and vecuronium were performed during a 90-minute infusion period and also during storage of one of the mixtures for 30 days at 4 degrees C, 22 degrees C, and 30 degrees C . Initial and storage pH were determined, and assays of the three drugs were performed when pH of the mixtures was varied between 1.4 and 12.7 . Emulsion stability was assessed by measuring immediate and delayed alterations in droplet charge (zeta potential) and size . Finally, samples of the stored mixtures were incubated in culture medium for one week . SETTING: Teaching hospital and university laboratories . MEASUREMENTS AND MAIN RESULTS: Concentrations of the three drugs remained unchanged during infusion . During storage, concentrations of all three drugs were stable for two weeks at 4 degrees C or 22 degrees C, and for several days at 30 degrees C . Initial pH of the two mixtures was 5.1