Int J Food Microbiol, 1999 Sep 15, 50(1-2), 25 - 31 The safety and social acceptance of novel foods; Moseley BE; The regulatory processes employed in the UK and the European Union to assess the safety of novel foods and novel food ingredients, including those resulting from the application of recombinant DNA technology (genetically modified foods), are described . Examples are given of yeasts that have been genetically modified and can be used in food and drink manufacture and food enzymes derived from genetically modified microorganisms that have been deemed safe for use by the UK regulatory system . Social acceptance of such novel foods or food ingredients is not uniform in countries of the developed world . Consumer concerns can be based on ethical considerations (scientists "playing God") or safety worries ("more testing needs to be done") . The general acceptance of such foods and food ingredients in Europe is still unclear. Cornea, 1999 Sep, 18(5), 608 - 11 The treatment of Pseudomonas keratoscleritis after pterygium excision; Huang SC et al.; PURPOSE: To assess the effect of intensive topical and intravenous antibiotics plus oral prednisolone and surgical debridement in Pseudomonas keratoscleritis after pterygium excision . METHODS: We describe three cases of P . aeruginosa-induced keratoscleritis occurring 10 days to 18 months after uncomplicated pterygium excision . Treatment included early conjunctival debridement, topical and intravenous antibiotics, and low-dosage oral prednisolone . RESULTS: All three patients responded to the combined therapy . Microorganisms were eliminated, and ulcers were healed within 8 weeks . Treatment was not extended beyond that, and infection did not recur . No evisceration was required . The patients' best corrected visual acuities are 20/200, 20/400, and 20/120, respectively . CONCLUSION: Early diagnosis and prompt, intensive medical and surgical treatment may save a patient's vision and forestall evisceration. Vestn Ross Akad Med Nauk, 1999, (8), 43 - 7 {The group identification of the causative agents of infectious diseases in environmental objects}; Buianov VV et al.; Drop colorimetric reactions were studied for group identification of microorganisms referring to as viral rickettsial bacterial vegetative and bacterial sporal taxonomic groups . For their determination, the authors proposed a number of well-known reactions for preliminary detection of microorganisms if there was protein or protein components in a sample, as well as a series of reactions characteristic of some groups of microorganisms and their vital activity . The list was proposed to include not only tests for dehydrogenase and peroxidase activities and for inorganic phosphates, but also those for the ability of suspension production, etc, which were specific signs of the presence and vital ability of microorganisms . Drop reactions were made with eight indicator papers and a set of reagents used for control of the reactions, and with a set of wetting solutions for liquid-phase reactions . A set of reagents and accessories provided the identification of 40 samples of microorganisms, the analysis of one unknown sample took less than an hour . The authors developed an algorithm for the sequence of tests depending on the combination of positive and negative effects of indicator means, namely an algorithm for relating microorganisms to the above taxonomic groups. Curr Microbiol, 1999 Oct, 39(4), 200 - 4 Tryptophan biosynthesis and production of other related compounds from indole and L-serine by mixed ruminal bacteria, protozoa, and their mixture in vitro; Mohammed N et al.; Tryptophan (Trp) biosynthesis and production of other related compounds from 1 mM each of indole (IND), L-serine (Ser), and IND plus Ser by mixed ruminal bacteria (B), protozoa (P), and their mixture (BP) in an in vitro system were quantitatively investigated . Ruminal microorganisms were anaerobically incubated at 39 degrees C for 12 h . Trp and other related compounds produced in both the supernatants and microbial hydrolyzates of the incubation were analyzed by HPLC . B, P, and BP suspensions were not able to produce Trp when incubated with only IND or Ser . Appreciable amounts of Trp (9.8, 3.1, and 6.6% of substrate) were produced from IND plus Ser after 12 h by B, P, and BP suspensions, respectively . Trp produced from IND + Ser in B was found only in the hydrolyzate, whereas it was found in the medium as a free form in P after a 12-h incubation period . Rumen bacteria and protozoa were separately demonstrated for the first time to produce Trp from IND plus Ser, and the ability of P to produce Trp from IND plus Ser was about one-third that of B in 12 h . Trp produced from IND plus Ser by B, P, and BP suspensions was simultaneously degraded into its related compounds, and, among them, indoleacetic acid (IAA) was a major product found in B . Production of IAA was 4.3, 0.3, and 3.2% of IND in 12 h by B, P, and BP suspensions, respectively . A small amount of skatole (SKT) (1.1 and 2.5% in B and BP, respectively) and p-cresol (CRL) (2.4 and 3.4% in B and BP, respectively) were also produced from IND plus Ser during 12-h incubation . P suspension produced no SKT or CRL from IND plus Ser in 12-h incubation . These results suggested for the first time that both rumen bacteria and protozoa have an ability to produce Trp from IND plus Ser, and the ability was higher in B than in P . The ratios of Trp produced from IND plus Ser to that from indolepyruvic acid by B, P, and BP were 1:3.4, 1:14.2, and 1:6.6 during 12-h incubation period . From these results, the degree of importance of producing Trp from IND plus Ser in the rumen was indicated. Biochim Biophys Acta, 1999 Aug 4, 1412(3), 191 - 211 Does microbial life always feed on negative entropy? Thermodynamic analysis of microbial growth; von Stockar U et al.; Schrodinger stated in his landmark book, What is Life?, that life feeds on negative entropy . In this contribution, the validity of this statement is discussed through a careful thermodynamic analysis of microbial growth processes . In principle, both feeding on negative entropy, i.e . yielding products of higher entropy than the substrates, and generating heat can be used by microorganisms to rid themselves of internal entropy production resulting from maintenance and growth processes . Literature data are reviewed in order to compare these two mechanisms . It is shown that entropy-neutral, entropy-driven, and entropy-retarded growth exist . The analysis of some particularly interesting microorganisms shows that enthalpy-retarded microbial growth may also exist, which would signify a net uptake of heat during growth . However, the existence of endothermic life has never been demonstrated in a calorimeter . The internal entropy production in live cells also reflects itself in the Gibbs energy dissipation accompanying growth, which is related quantitatively to the biomass yield . An empirical correlation of the Gibbs energy dissipation in terms of the physico-chemical nature of the growth substrate has been proposed in the literature and can be used to predict the biomass yield approximately . The ratio of enthalpy change and Gibbs energy change can also be predicted since it is shown to be approximately equal to the same ratio of the relevant catabolic process alone. J Gastrointest Surg, 1999 Sep-Oct, 3(5), 496 - 505 Preoperative biliary drainage: impact on intraoperative bile cultures and infectious morbidity and mortality after pancreaticoduodenectomy; Povoski SP et al.; Whether it is necessary to perform biliary drainage for obstructive jaundice before performing pancreaticoduodenectomy remains controversial . Our aim was to determine the impact of preoperative biliary drainage on intraoperative bile cultures and postoperative infectious morbidity and mortality following pancreaticoduodenectomy . We retrospectively analyzed 161 consecutive patients undergoing pancreaticoduodenectomy in whom intraoperative bile cultures were performed . Microorganisms were isolated from 58% of these intraoperative bile cultures, with 70% of them being polymicrobial . Postoperative morbidity was 47% and mortality was 5% . Postoperative infectious complications occurred in 29%, most commonly wound infection (14%) and intra-abdominal abscess (12%) . Eighty-nine percent of patients with intra-abdominal abscess (P = 0.003) and 87% with wound infection (P = 0.003) had positive intraoperative bile cultures . Microorganisms in the bile were predictive of microorganisms in intraabdominal abscess (100%) and wound infection (69%) . Multivariate analysis of preoperative and intraoperative variables demonstrated that preoperative biliary drainage was associated with positive intraoperative bile cultures (P <0.001), postoperative infectious complications (P = 0.022), intra-abdominal abscess (P = 0.061), wound infection (P = 0.045), and death (P = 0 . 021) . Preoperative biliary drainage increases the risk of positive intraoperative bile cultures, postoperative infectious morbidity, and death . Positive intraoperative bile cultures are associated with postoperative infectious complications and have similar microorganism profiles . These data suggest that preoperative biliary drainage should be avoided in candidates for pancreaticoduodenectomy. Trends Biotechnol, 1999 Oct, 17(10), 403 - 9 The Earth's bounty: assessing and accessing soil microbial diversity; Rondon MR et al.; The study of microbial diversity represents a major opportunity for advances in biology and biotechnology . Recent progress in molecular microbial ecology shows that the extent of microbial diversity in nature is far greater than previously thought . Here, we discuss methods to analyse microorganisms from natural environments without culturing them and new approaches for gaining access to the genetic and chemical resources of these microorganisms. Rev Esp Med Nucl, 1999 Aug, 18(4), 272 - 5 {Validation of the preparation of individual doses of 51Cr-EDTA in syringes}; Iglesias Allende F et al.; The 51Cr-EDTA is one of the radiopharmaceuticals more used in the glomerular filtration rate determination . The necessity to prepare a daily reference dose every time that are prepared the patient's doses can go to a poor exploitation of the multidose vial . One way of improving its use consists on the preparation of individual doses in ready syringes to inject, for several days, employing only one of them as a standard . The aim of this study was to evaluate the storing of 51Cr-EDTA individual doses at room temperature and at 4 degrees C during 4 weeks, analyzing the radiochemical purity, the sterility and the radiopharmaceutical retention into the syringe . The results obtained showed that the radiochemical purity remains stable, always been greater than the value recommended for its use (> 95%) . Likewise, there were not microorganism contamination nor 51Cr-EDTA appreciable retention into the syringes . We concluded that the preparation of individual doses of 51Cr-EDTA in syringes and their conservation at room temperature, or at 4 degrees C, during a period of 4 weeks, neither influence in the radiopharmaceutical quality nor increase its retention into the syringe. Tidsskr Nor Laegeforen, 1999 Aug 10, 119(18), 2670 - 3 {Use of short wave ultraviolet radiation for disinfection in operating rooms}; Banrud H et al.; Over a number of years, short wave ultraviolet radiation (UVC; 200-280 nm) has been used to disinfect air and surfaces in operating rooms, patient rooms, laboratories and so on, as well as air in ventilation ducts . Despite the well-documented effect of ultraviolet radiation on air quality, thus reducing the occurrence of infections, this technology has been relatively little used . One advantage of this method is that the UVC sources ensure a continuous reduction in the number of airborne microorganisms that are generated all the time . There are, however, some disadvantages with this method . Human exposure to ultraviolet C may cause keratoconjunctivitis and erythema and requires protection of the skin and the eyes of people exposed to levels above recommended exposure limits . However, by enclosing the UVC sources or by irradiating in the absence of human activity, human exposure is eliminated . These and other aspects concerning the use of short wave ultraviolet radiation as a disinfection agent in operating rooms are discussed in this article. J Biol Chem, 1999 Sep 17, 274(38), 27105 - 11 Purification and characterization of phosphopantetheine adenylyltransferase from Escherichia coli; Geerlof A et al.; Phosphopantetheine adenylyltransferase (PPAT) catalyzes the penultimate step in coenzyme A (CoA) biosynthesis: the reversible adenylation of 4'-phosphopantetheine yielding 3'-dephospho-CoA and pyrophosphate . Wild-type PPAT from Escherichia coli was purified to homogeneity . N-terminal sequence analysis revealed that the enzyme is encoded by a gene designated kdtB, purported to encode a protein involved in lipopolysaccharide core biosynthesis . The gene, here renamed coaD, is found in a wide range of microorganisms, indicating that it plays a key role in the synthesis of 3'-dephospho-CoA . Overexpression of coaD yielded highly purified recombinant PPAT, which is a homohexamer of 108 kDa . Not less than 50% of the purified enzyme was found to be associated with CoA, and a method was developed for its removal . A steady state kinetic analysis of the reverse reaction revealed that the mechanism of PPAT involves a ternary complex of enzyme and substrates . Since purified PPAT lacks dephospho-CoA kinase activity, the two final steps of CoA biosynthesis in E . coli must be catalyzed by separate enzymes. Transplantation, 1999 Aug 27, 68(4), 535 - 40 Increased incidence of postoperative infections associated with peritoneal dialysis in renal transplant recipients; Passalacqua JA et al.; BACKGROUND: Infection is a frequent postoperative complication in renal transplant recipients . However, little information is available concerning the effect of pretransplantation dialysis modality on posttransplantation complications including infection . We therefore evaluated the effect of hemodialysis (HD) versus peritoneal dialysis (PD) on the incidence of postoperative infection as well as several other posttransplantation outcomes . METHODS: A retrospective analysis was performed using medical records covering the period 30 days after transplantation of 156 dialysis patients who underwent renal transplantation at a single center during a 22-month period . Of these patients, 103 received only HD, 32 received only PD, 13 received PD in the past and HD immediately before transplantation (PH/HD), and 8 received HD in the past and PD immediately before transplantation (HD/PD) . The presence of culture-proven infection, types of infecting organisms, length of initial hospital stay, and incidence of rejection during the first 30 days after transplantation were determined for each patient . RESULTS: All groups were similar with regard to age, race, gender, underlying disease, donor type, incidence of delayed graft function, and perioperative antibiotic prophylaxis . There were more infectious complications within 30 days after transplantation in patients on PD just prior to transplantation (PD and HD/PD) than in HD patients (67.5% vs . 25.9%, P<0.00001) . When types of infectious organisms were assessed, PD patients were found to have a greater incidence of infections with microorganisms that colonize human skin (P<0.0001) . The median length of hospital stay was 3 days longer for PD patients and 6.5 days longer for HD/PD patients than for patients receiving HD (P=0.01 and 0.04), and PD and HD/PD patients were more likely to have an episode of rejection than HD patients (P=0.02) . CONCLUSIONS: Renal replacement therapy with PD immediately before transplantation negatively affects outcome as compared with HD, predisposing patients to a greater incidence of postoperative infections and rejection and a longer hospital stay . Further study in a randomized controlled trial may help determine how adjustment of the dialysis method can optimize transplantation outcome. J Microbiol Methods, 1999 Sep, 37(3), 245 - 53 A PCR test for the identification and discrimination of Legionella longbeachae serogroups 1 and 2; Saint CP et al.; A PCR test has been developed for the specific identification of Legionella longbeachae . The test targeted sequence unique to both L . longbeachae serogroups 1 and 2 within the mip gene and permitted both species and serogroup identification . The test was trialed on a range of closely related species and 20 clinical isolates originating from Australia, the USA and Israel . Results were consistent with previous identification analyses . From 20 water samples known to contain Legionella spp . one sample yielded isolates which consistently tested positive by L . longbeachae serogroup 1 PCR . DNA sequencing of the PCR product, 5S rRNA gene sequence and hybridisation analysis with a specific oligonucleotide probe definitively identified one isolate as L . longbeachae serogroup 1 . PCR testing was demonstrated as a superior method of identification to traditional seroagglutination reactions, which were ambiguous and could explain the previous failure to identify the presence of this microorganism in water. J Microbiol Methods, 1999 Sep, 37(3), 201 - 13 In situ imaging of microorganisms in geologic material; Tobin KJ et al.; In order to fully delineate the interactions of microorganisms with geological substrates, unequivocal identification of intact microbial cells within geologic samples is required without the disruption of either the rock texture or the relationship of the microorganisms to the mineral fabric . To achieve this objective we developed a protocol that enables the visualization of intact microbial cells in petrographic thin sections, avoids detaching the cells from their host mineral surfaces and avoids microbial contamination during the lapidary process . Propidium iodide and POPO-3, nucleic acid stains that specifically target double-stranded DNA and RNA were utilized for in situ visualization of cells in surface and subsurface basalts from northeastern Idaho . Additionally, examination of samples incubated with acetic acid-UL-14C via phosphor imagining facilitated the in situ visualization of 14C labeled biomass . Biomass observed was low (<10(7) cells/g) . These observations indicate that the microbial distribution in these rocks exhibits a high degree of spatial heterogeneity at the sub-centimeter scale. Rev Esp Enferm Dig, 1999 Jul, 91(7), 508 - 15 {Are all H . pylori "bad"?}; Gisbert J et al.; The infection by Helicobacter pylori arises a question of great interest: should the infection be considered a disease in all cases? or, on the contrary, is the microorganism harmful in only some circumstances? The response to these questions is followed by therapeutic implications of great importance, as it is followed by the decision of administering eradication therapy to every patient or only to some individuals . In this article the arguments supporting each of the aforementioned options are reviewed . Firstly, the aphorism recently stated <<The only good H . pylori is a dead H . pylori>> suggests that all microorganisms are harmful and, therefore, generalized H . pylori eradication therapy should be administered to all infected subjects . Among the arguments supporting this attitude are the following: the H . pylori <<healthy>> carrier status does not exist; a high risk of developing peptic ulcer exists in infected patients; H . pylori positive patients are at risk of suffering gastric adenocarcinoma and lymphoma; and, finally, the eradication of the microorganism would reduce the potential reservoir of the infection . On the other hand, the expression <<not all H . pylori strains are created equal: should all be eliminated?>> summarises a more conservative position, that suggests the existence of <<bad>>, <<neutral, or even <<good>> H . pylori, advising the eradication of the microorganism in only some cases . Several arguments supporting this idea, and, therefore, against the generalized use of eradication therapy, have been proposed: H . pylori has colonized humans since immemorial time and, therefore, it can not be very harmful to the human; the bacterium will induce peptic ulcer or gastric cancer in only a minority of infected subjects; the beneficial effect of eradication has been definitively demonstrated in few entities as gastroduodenal ulcer and MALT lymphoma; some strains could be beneficial to the human; generalized administration of antibiotic therapy will cause problems; the H . pylori vaccine represents a more rational alternative; and, finally, the incidence of H . pylori infection is spontaneously decreasing in some regions . In summary, there are strong arguments supporting both of the strategies that are reviewed in the present article, and, therefore, the dilemma <<good H . pylori, bad H . pylori>> remains. Parasite Immunol, 1999 Sep, 21(9), 461 - 73 Vaccination against Leishmania major in a CBA mouse model of infection: role of adjuvants and mechanism of protection; Rivier D et al.; Gp63 is a major surface protein of Leishmania promastigotes . Its protective efficacy has been tested in several experimental models using different mouse strains, gp63 forms, adjuvants and routes of immunization, giving rise to conflicting results . This investigation was designed to determine whether these discrepancies could be ascribed to differing experimental procedures, and to compare gp63-induced protection with that achieved using live promastigotes . Preliminary experiments demonstrated that gp63 was an extremely potent immunogen compared to a standard antigen (ovalbumin) . Protection against Leishmania major infection afforded by gp63 inoculation was studied in CBA mice . Injection of gp63 in saline, or of CFA, BCG, and C . parvum without antigen, induced significant protection . When gp63 and adjuvants were combined, results differed depending on the site of vaccination relative to that of the challenge infection . Vaccination with gp63 plus adjuvants in the tail (i.e . close to the site of infection) led to a stronger reduction of lesion size than the basal level of protection elicited by adjuvants alone, except in the case of CFA . Surprisingly however, when the antigen was injected at a distance from the site of infection (immunization in the hind foot pads, infection in the rump), the protective effect of gp63 was decreased by the adjuvants . Finally, vaccination at either site using live parasites (radioattenuated or virulent promastigotes) resulted in most instances in better protection than achieved by any protocol using gp63 and adjuvants . While anti-gp63 T cells proliferated in vitro in response to L . major-infected bone marrow-derived macrophages, they were unable to activate macrophages for parasite killing . This is in contrast with lymphocytes from mice immunized with live parasites, which both proliferated and stimulated significant killing of the microorganisms within 48 h. Mol Immunol, 1999 Jun, 36(8), 505 - 14 Gene structure of the P100 serine-protease component of the human Ra-reactive factor; Takayama Y et al.; The Ra-reactive factor (RaRF) is a complement dependent anti-microbial factor that reacts with numerous microorganisms such as viruses, bacteria, fungi and protozoa . It is a complex of a mannan-binding lectin (MBL) and the serine protease, P100 (MASPI) . P100 activates the C4 component of the complement system and its domain organization is similar to C1r and C1s . In this study, determination was made of the structure of the human P100 gene which was found longer than 67 kbp and to be comprised of 16 exons . Its non-protease region consisted of 10 exons, as in the case of C1r and C1s, and the introns were found present in the boundary separating two CUB domains, an EGF-like domain and two CCP domains and each CUB and CCP domain contained extra internal introns . The serine protease region was comprised of 6 exons in contrast to C1r and C1s, either of which consists of a single exon . The exon-intron structure was found to reflect the evolution of these molecules and P100 to have derived earlier in the stage of evolution than C1r or C1s. In Vitro Cell Dev Biol Anim, 1999 Jan, 35(1), 43 - 8 Lipopolysaccharide of Escherichia coli, polyamines, and acetic acid stimulate cell proliferation in intestinal epithelial cells; Olaya J et al.; Our aim was to examine whether lipopolysaccharide of Escherichia coli, polyamines of dietetic and/or bacterial origin, and products of the bacterial metabolism influence cell proliferation in epithelial cells from the colon and small intestine . Lipopolysaccharide of Escherichia coli 0111:B4 was incubated with cultures from human colonic mucosa . The mitoses were arrested with Vincristine and the total number of metaphases per crypt was counted . In addition, lipopolysaccharide was incubated with a human colonic epithelial cell line from adenocarcinoma (LS-123 cells) and with a nontransformed small intestinal cell line from germ-free rats (IEC-6 cells) for 24 h . In the last 4 h, the cells were labeled with tritiated thymidine . The cells were incubated with putrescine, cadaverine, and spermidine at 10(-11)-10(-3) M and with acetic acid (10(-5)-10(-1) M), acetaldehyde (10(-10)-10(-4) M) and ammonium chloride (1-20 mM) . Lipopolysaccharide of Escherichia coli increased the number of arrested metaphases in human colonic crypts and DNA synthesis in L-123 and IEC-6 cells (P < 0.001) . All polyamines increased DNA synthesis in the colonic and small intestinal cell lines, the effects being more marked for putrescine (P < 0.001) . The higher concentrations of acetic acid increased DNA synthesis in both epithelial cell lines (P < 0.001) . Acetaldehyde slightly decreased DNA synthesis in LS-123 cells at cytotoxic concentrations . Ammonium chloride did not significantly affect DNA synthesis . The final concentration of nonionized ammonia was less than 3% . It is concluded that lipopolysaccharides of Escherichia coli and intraluminal factors derived from microorganisms increase cell proliferation in human colonic crypts and intestinal epithelial cell lines. Yakugaku Zasshi, 1999 Aug, 119(8), 529 - 83 {Antitumor compounds isolated from higher plants}; Itokawa H et al.; A lot of anticancer agents have been isolated from natural sources, especially from microorganisms and plants . However, there is no special type of compounds for cancer therapy . Various types of substances are effective for various types of cancers and tumors: for instance, alkaloids, lignans, terpenes and steroids, etc . In this report, the authors will describe especially about higher plants. Zentralbl Chir, 1999, 124(7), 573 - 84 {Parenteral and enteral nutrition in palliative medicine}; Herrmann A et al.; Artificial nutritional support does not alter the natural course in patients with malignant disease . The outcome of these patients is mainly determined by the type and stage of the underlying tumor . Progress of the underlying disease is often paralleled by malnutrition which in turn facilitates complications and may reduce survival and quality of life . Nutritional support can be applied to maintain body weight, immune function and quality of life . Enteral nutrition can be applied with a functioning gastrointestinal tract and has been proven to be superior compared to parenteral nutrition . Maintenance of intestinal mucosal function due to enteral substrate application prevents disruption of intestinal barrier function as well as the overgrowth of intestinal microorganisms . Using a step-by-step approach dietetic counselling in combination with augmented oral caloric intake should be the first measure . The next step to take is nutritional support by enteral tube feeding using formula diets . Parenteral feeding should only be used if other options to support caloric intake have failed. Appl Environ Microbiol, 1999 Sep, 65(9), 4064 - 70 Radioactive fingerprinting of microorganisms that oxidize atmospheric methane in different soils; Roslev P et al.; Microorganisms that oxidize atmospheric methane in soils were characterized by radioactive labelling with (14)CH(4) followed by analysis of radiolabelled phospholipid ester-linked fatty acids ((14)C-PLFAs) . The radioactive fingerprinting technique was used to compare active methanotrophs in soil samples from Greenland, Denmark, the United States, and Brazil . The (14)C-PLFA fingerprints indicated that closely related methanotrophic bacteria were responsible for the oxidation of atmospheric methane in the soils . Significant amounts of labelled PLFAs produced by the unknown soil methanotrophs coeluted with a group of fatty acids that included i17:0, a17:0, and 17:1omega8c (up to 9.0% of the total (14)C-PLFAs) . These PLFAs are not known to be significant constituents of methanotrophic bacteria . The major PLFAs of the soil methanotrophs (73.5 to 89.0% of the total PLFAs) coeluted with 18:1 and 18:0 fatty acids (e.g., 18:1omega9, 18:1omega7, and 18:0) . The (14)C-PLFAs fingerprints of the soil methanotrophs that oxidized atmospheric methane did not change after long-term methane enrichment at 170 ppm CH(4) . The (14)C-PLFA fingerprints of the soil methanotrophs were different from the PLFA profiles of type I and type II methanotrophic bacteria described previously . Some similarity at the PLFA level was observed between the unknown soil methanotrophs and the PLFA phenotype of the type II methanotrophs . Methanotrophs in Arctic, temperate, and tropical regions assimilated between 20 and 54% of the atmospheric methane that was metabolized . The lowest relative assimilation (percent) was observed for methanotrophs in agricultural soil, whereas the highest assimilation was observed for methanotrophs in rain forest soil . The results suggest that methanotrophs with relatively high carbon conversion efficiencies and very similar PLFA compositions dominate atmospheric methane metabolism in different soils . The characteristics of the methane metabolism and the (14)C-PLFA fingerprints excluded any significant role of autotrophic ammonia oxidizers in the metabolism of atmospheric methane. J Ethnopharmacol, 1999 Sep, 66(3), 289 - 93 Screening of Turkish anti-ulcerogenic folk remedies for anti-Helicobacter pylori activity; Yesilada E et al.; The anti-Helicobacter pylori effect of the extracts and fractions obtained from seven Turkish plants, which are used in folk medicine for the treatment of gastric ailments including peptic ulcers, were studied against one standard strain and eight clinical isolates of H . pylori by using the agar dilution method . Flowers of Cistus laurifolius and Spartium junceum, cones of Cedrus libani, herbs and flowers of Centaurea solstitialis ssp . solstitialis, fruits of Momordica charantia, herbaceous parts of Sambucus ebulus, and flowering herbs of Hypericum perforatum were evaluated in this study . Results showed that all except one extract from six of these plants showed activity against the microorganism with MICs between 1.95 and 250 microg/ml, with S . junceum being the only inactive species . Amongst the active plants the inhibitory properties of C . laurifolius were found prominent. J Clin Immunol, 1999 Jul, 19(4), 203 - 14 New insights into the biology of the acute phase response; Suffredini AF et al.; Innate or natural immunity is a highly conserved defense mechanism against infection found in all multicellular organisms . The acute phase response is the set of immediate inflammatory responses initiated by pattern recognition molecules . These germ cell-encoded proteins recognize microbial pathogens based on shared molecular structures and induce host responses that localize the spread of infection and enhance systemic resistance to infection . Innate immunity also influences the initiation and type of adaptive immune response by regulating T cell costimulatory activity and antigen presentation by antigen presenting cells and by influencing mediator production, which affects lymphocyte function and trafficking . Acute phase protein concentrations rapidly increase after infection, and their production is controlled primarily by IL-6- and IL-1-type cytokines . The acute phase proteins provide enhanced protection against microorganisms and modify inflammatory responses by effects on cell trafficking and mediator release . For example, serum amyloid A has potent leukocyte activating functions including induction of chemotaxis, enhancement of leukocyte adhesion to endothelial cells, and increased phagocytosis . The constellation of inflammatory responses seen after endotoxin administration to humans represents an in vivo model of the acute phase response . Studies with inflammatory modifying agents, such as soluble dimeric TNF receptor and IL-10, show that these responses are not dependent on a single mediator but result from multiple overlapping inflammatory pathways . Understanding the factors that initiate and alter the magnitude and duration of the acute phase response represents an important step in the development of new therapies for infectious and inflammatory diseases. Arch Oral Biol, 1999 Sep, 44(9), 711 - 9 Inhibition of experimental bone resorption and osteoclast formation and survival by 2-aminoethanesulphonic acid; Koide M et al.; It is known that bone resorption is mediated by osteoclasts, and lipopolysaccharide (LPS) and inflammatory mediators such as interleukin-1 (IL-1) and prostaglandin E2 (PGE2) induce osteoclast differentiation from haemopoietic cells, 2-aminoethanesulphonic acid, which is known as taurine, is an important nutrient and is added to most synthetic human infant milk formulas . In this study, it was found that 2-aminoethanesulphonic acid inhibits the stimulation of bone resorption mediated by LPS of the periodontopathic microorganism Actinobacillus actinomycetemcomitans Y4 in organ cultures of newborn mouse calvaria . The effect of 2-aminoethanesulphonic acid on the development and survival of osteoclast-like multinucleated cells produced in a mouse bone-marrow culture system was also examined . 2-aminoethanesulphonic acid (100 microg/ml) suppressed the formation of these osteoclast-like cells in the presence of LPS of A . actinomycetemcomitans Y4, IL-1alpha or PGE2 in mouse marrow cultures . On the other hand, 2-aminoethanesulphonic acid did not inhibit 1alpha, 25-dihydroxyvitamin D3-mediated osteoclast differentiation . Although IL-1alpha elongated the survival of the osteoclast-like cells, 2-aminoethanesulphonic acid blocked the supportive effect of IL-1alpha on osteoclast survival . 2-aminoethanesulphonic acid showed no effect on the growth of mouse osteoblasts . Finally, it was found that 2-aminoethanesulphonic acid inhibited alveolar bone resorption in experimental periodontitis in hamsters . These results suggest that 2-aminoethanesulphonic acid is an effective agent in preventing inflammatory bone resorption in periodontal diseases. EMBO J, 1999 Sep 1, 18(17), 4733 - 43 Steady-state free Ca(2+) in the yeast endoplasmic reticulum reaches only 10 microM and is mainly controlled by the secretory pathway pump pmr1; Strayle J et al.; Over recent decades, diverse intracellular organelles have been recognized as key determinants of Ca(2+) signaling in eukaryotes . In yeast however, information on intra-organellar Ca(2+) concentrations is scarce, despite the demonstrated importance of Ca(2+) signals for this microorganism . Here, we directly monitored free Ca(2+) in the lumen of the endoplasmic reticulum (ER) of yeast cells, using a specifically targeted version of the Ca(2+)-sensitive photoprotein aequorin . Ca(2+) uptake into the yeast ER displayed characteristics distinctly different from the mammalian ER . At steady-state, the free Ca(2+) concentration in the ER lumen was limited to approximately 10 microM, and ER Ca(2+) sequestration was insensitive to thapsigargin, an inhibitor specific for mammalian ER Ca(2+) pumps . In pmr1 null mutants, free Ca(2+) in the ER was reduced by 50% . Our findings identify the secretory pathway pump Pmr1, predominantly localized in the Golgi, as a major component of ER Ca(2+) uptake activity in yeast. Br J Dermatol, 1999 Aug, 141(2), 240 - 5 Development of an oligonucleotide probe specific for Trichophyton rubrum; El Fari M et al.; A species-specific DNA probe was developed to detect the dermatophyte species Trichophyton rubrum . The selected oligonucleotide sequence is derived from the highly variable internal transcribed spacer 2 region of the ribosomal DNA operon . The specificity of the non-radioactive labelled oligonucleotide probe was tested against related dermatophytes, other eukaryotic microorganisms and against human DNA . No cross-hybridization was found, and hybridization signals were invariably detected in all T . rubrum strains investigated . In addition, no homologous sequences were found searching the EMBL database . Experiments to establish a method for isolating DNA directly from clinical specimens gave successful amplification and hybridization products in about 30% of the samples. Biol Trace Elem Res, 1999 Sep, 69(3), 217 - 34 Accelerated volatilization rates of selenium from different soils; Stork A et al.; Selenium (Se), an element found naturally in a variety of soils, can accumulate in drainage water of lands under intensive irrigation, even reaching levels that are toxic to mammals and birds . Volatilization of Se by soil microorganisms into dimethylselenide (DMSe) can be enhanced by certain soil amendments and, thus, be used as a soil remediation process . In an 8-wk laboratory study, five soils from California and one from Germany were spiked with 75SeO3(2-) (22.3 mg/kg Se) . Two amino acids (DL-homocysteine and L-methionine), a carbohydrate (pectin), and a protein (zein) were tested as soil amendments . Gaseous 75Se emissions were trapped with activated carbon and measured in a gamma counter . Depending on soil type, the cumulative volatilization from the control flasks varied between 1.2% and 9.0% of applied 75Se . Both zein and L-methionine strongly increased volatilization (max . 43% of 75Se applied), whereas DL-homocysteine had a much smaller stimulating effect . Pectin showed a moderate effect, but enhanced Se volatilization rates were sustained much longer when compared to the zein amendment . Volatilization rates of Se followed a simple first-order reaction . Gaseous Se emission in the soils treated with L-methionine yielded an S-shaped curve, which fit a growth-modified first-order rate model . Although zein and L-methionine were the most favorable treatments enhancing Se volatilization, all six soils responded differently to the soil amendments. Proc Natl Acad Sci U S A, 1999 Aug 31, 96(18), 10489 - 94 Mitomycin resistance in mammalian cells expressing the bacterial mitomycin C resistance protein MCRA; Belcourt MF et al.; The mitomycin C-resistance gene, mcrA, of Streptomyces lavendulae produces MCRA, a protein that protects this microorganism from its own antibiotic, the antitumor drug mitomycin C . Expression of the bacterial mcrA gene in mammalian Chinese hamster ovary cells causes profound resistance to mitomycin C and to its structurally related analog porfiromycin under aerobic conditions but produces little change in drug sensitivity under hypoxia . The mitomycins are prodrugs that are enzymatically reduced and activated intracellularly, producing cytotoxic semiquinone anion radical and hydroquinone reduction intermediates . In vitro, MCRA protects DNA from cross-linking by the hydroquinone reduction intermediate of these mitomycins by oxidizing the hydroquinone back to the parent molecule; thus, MCRA acts as a hydroquinone oxidase . These findings suggest potential therapeutic applications for MCRA in the treatment of cancer with the mitomycins and imply that intrinsic or selected mitomycin C resistance in mammalian cells may not be due solely to decreased bioactivation, as has been hypothesized previously, but instead could involve an MCRA-like mechanism. J Mal Vasc, 1999 Jun, 24(3), 194 - 201 {Infections of vascular prostheses of the abdominal aorta . Diagnostic and therapeutic problems}; Parola P et al.; Aortic graft infection is one of the most serious complication of vascular surgery . While the incidence of such infections has declined over the years, it continues to be associated with high mortality and high rates of limb loss . Graft implanted in the inguinal area have a higher rate of infection than those that lie entirely within the abdomen . Infection of the intraabdominal extremities of vascular grafts is most frequently associated with prostheto-enteric fistula . DIAGNOSIS AND TREATMENT: Despite the many available sophisticated imaging techniques diagnosis remains difficult, particularly with intra-abdominal grafts because of nonspecific clinical and imaging findings . A variety of approaches to aortic graft infection have been proposed and optimal treatment usually requires an association of systemic antibiotic administration with surgery . Removal of the entire infected graft is often necessary for cure and there are only few situations in which conservative treatment is acceptable . A careful identification of the infecting microorganisms and in vitro susceptibility testing are essential for successful therapy . PREVENTION: Prevention consists in a strict adherence to principles of asepsis and the use of prophylactic antibiotics in vascular surgery . The use of antibiotics-bound knitted grafts has been recently proposed. Immunopharmacol Immunotoxicol, 1999 Aug, 21(3), 421 - 38 The effects of HIV viral load on the phagocytic activity of monocytes activated with lipopolysaccharide from oral microorganisms; Baqui AA et al.; A study was undertaken to determine whether viral load status in HIV+ patients has any potential effect on monocyte phagocytic function both before and after challenge of the monocytes with lipopolysaccharide (LPS) isolated from oral microorganisms . LPS of two putative periodontal pathogens Porphyromonas gingivalis (P . gingivalis) and Fusobacterium nucleatum (F . nucleatum) was prepared . Whole blood samples in EDTA were collected from 30 HIV+ patients presenting for dental care at the University of Maryland . Control samples were prepared from appropriate uninfected individuals . Viral load was determined using quantitative RT-PCR (Amplicor, Roche Diagnostics) . Phagocytic function was determined using FITC labeled Saccharomyces species in resting isolated monocytes and in cells after 24 h stimulation with 1 microgram/ml of LPS of P . gingivalis or F . nucleatum . Immunohistochemical staining was performed for complement receptor CR-1 (CD-35) on phagocyte cells . In HIV+ patients with high viral load (> 10,000 copies/ml), 13.5% of isolated resting monocytes demonstrated phagocytic activity, while 23% of the resting control monocytes from non-infected individuals showed phagocytic function . When the monocytes were stimulated with 1 microgram/ml of LPS of F . nucleatum, phagocytic activity was observed in 18.5% of monocytes in patients with high viral load, 33.5% with moderate viral load (400-10,00 copies/ml) and 51% with low viral load (<400 copies/ml), while 62% of the control monocytes demonstrated phagocytic activity . Stimulation of monocytes with LPS of P . gingivalis showed similar results . Complement receptor CD-35 showed a 50% decrease in expression in HIV+ patients with high viral load . A progressive decrease in monocyte/macrophage phagocytic function and CD-35 expression with and without oral LPS activation occurs after HIV infection and this trend appears to be accentuated in patients with high viral load . This relationship may contribute to increased susceptibility to oral opportunistic infections in advanced HIV+ patients. Biodegradation, 1999 Apr, 10(2), 149 - 57 Analyses of polycyclic aromatic hydrocarbon-degrading bacteria isolated from contaminated soils; Ahn Y et al.; Polycyclic aromatic hydrocarbon (PAH)-degrading bacteria isolated from PAH-contaminated soils were analyzed genotypically and phenotypically for their capacity for metabolism of naphthalene and other PAH substrates . The methods used for the analyses were DNA hybridization using NAH7-derived gene probes, PAH spray plate assays, 14C-PAH mineralization assays, and dioxygenase activity assays . The results of the analyses showed a dominant number of PAH-degrading bacteria with a NAH7-like genotype . The results support the continued use of the nahA probe for contaminated soils to monitor the genetic potential of indigenous microorganisms to degrade PAHs . However, the finding of non-nahA-hybridizing PAH-degrading bacteria show the limitation of NAH7-derived gene probes . Fifteen percent (13/89) of PAH-degrading bacteria isolated were not detected with the nahA gene probe . Four isolates (designated A5PH1, A8AN3, B1PH2, and B10AN1) did not hybridize with any of the NAH7-derived gene probes (nahA, nahG, nahH, and nahR) used in this study . Considering the numerous unculturable microorganisms in nature and their potential genotypes, NAH7-derived gene probes may underestimate the microbial potential to catabolize PAHs . This necessitates development of new gene probes for enumeration and isolation of PAH-degrading bacteria to better understand the in situ microbial potential to degrade PAHs. Parasitology, 1999, 118 Suppl, S3 - 9 The utility of complete genome sequences in the study of pathogenic bacteria; Hood DW; The availability of complete genome sequences is a revolution in the study of microorganisms . A fully annotated genome sequence provides an interactive tool for scientists and influences the approach and focus of research . In this article I discuss the impact of genome sequencing projects of bacteria . Much useful data have been obtained but the experimental methods needed to fully exploit the information continue to develop . Some of the approaches and particular applications relevant to bacteria of clinical importance are discussed. J Food Prot, 1997 Jan, 60(1), 48 - 53 Characteristics of low-and high-fat beef patties: effect of high hydrostatic pressure; Carballo J et al.; The purpose of this study was to analyze the consequences of applying high pressures (100 and 300 MPa for 5 or 20 min) on characteristics such as water- and fat-binding properties, texture, color, microstructure, and microbiology of low-fat (9.2%) and high-fat (20.3%) beef patties . In nonpressurized patties, the low-fat product exhibited significantly poorer (P < 0.05) binding properties and higher (P < 0.05) Kramer shear force and Kramer energy than did high-fat patties . Although high pressure did not clearly influence the binding properties of low- and high-fat beef patties, it did produce a rise in the Kramer shear force and energy which were more pronounced at 300 MPa . High pressures altered patty color, the extent of alteration depending on fat content, pressure, and pressurizing time . Pressurizing high- and low-fat beef patties at 300 MPa not only produced a lethal effect (P < 0.05) on microorganisms, but caused sublethal damage as well. Braz J Med Biol Res, 1999 Sep, 32(9), 1073 - 6 McFarland nephelometer as a simple method to estimate the sensitivity of the polymerase chain reaction using Mycobacterium tuberculosis as a research tool; Bollela VR et al.; Polymerase chain reaction (PCR) has been widely investigated for the diagnosis of tuberculosis . However, before this technique is applied on clinical samples, it needs to be well standardized . We describe the use of McFarland nephelometer, a very simple approach to determine microorganism concentration in solution, for PCR standardization and DNA quantitation, using Mycobacterium tuberculosis as a model . Tuberculosis is an extremely important disease for the public health system in developing countries and, with the advent of AIDS, it has also become an important public health problem in developed countries . Using Mycobacterium tuberculosis as a research model, we were able to detect 3 M . tuberculosis genomes using the McFarland nephelometer to assess mycobacterial concentration . We have shown here that McFarland nephelometer is an easy and reliable procedure to determine PCR sensitivity at lower costs. J Anim Sci, 1999 Aug, 77(8), 2269 - 78 Effect of wheat and corn variety on fiber digestion in beef steers fed high-grain diets; Martin C et al.; Six Salers steers, fitted with ruminal and duodenal cannulas, were used in a double 3x3 Latin square design to assess the depressive effect of the nature of wheat, flint corn, and dent corn on fiber digestion in animals fed high-concentrate diets, and to determine the mechanisms involved in these negative digestive effects . Diets were balanced to be equal in starch content (47.7+/-2.3%) . The three cereals were characterized by ruminal starch digestibilities of 86.6, 60.8, and 34.8% for the wheat, dent corn, and flint corn, respectively . Ruminal digestion of NDF was lower with wheat- than with corn-based diets (49.4 vs . 55.2%; P<.001), and with dent corn than with flint corn (53 vs . 57.3%; P<.01) . Degradability of hay in nylon bags was not affected by the grain source in the diet (P>.1) . The mean retention time of forage particles in the rumen was similar between wheat and corn diets (P>.1), but it was lower for steers fed dent corn than for those fed flint corn (P<.05) . Most fibrolytic activities of the solid-associated microorganisms were lower (P<.05) in animals fed wheat than in those fed corn . Differences in fibrolytic activities of the solid-associated microorganisms between the two corn genotypes were not statistically significant (P>.1), but activities of all fibrolytic enzymes were lower (P<.05) with the dent than with the flint corn diet . Protozoal number in ruminal fluid was lower in animals receiving wheat than in those fed corn (177 vs . 789x10(3)/mL; P<.001) and was related to the high ruminal acidity (P<.01) of the wheat diet . Large modifications in the rumen microbial ecosystem between the two corn genotypes were not visible in protozoal numbers or pH . Total-tract digestion of NDF was the same for wheat and for corn diets, averaging 55% for the three diets . A postruminal compensation of NDF digestion (14% of the total tract NDF digestion) seemed to occur with the wheat diet . The lack of any postruminal NDF digestion (0%) with the two corn diets may suggest negative digestive interactions in the hindgut similar to those in the rumen. Appl Microbiol Biotechnol, 1999 Jul, 52(1), 49 - 56 Processes of liquefaction/solubilization of Spanish coals by microorganisms Laborda F, Monistrol IF, Luna N, Fernandez M. Several fundamental aspects of microbial coal liquefaction/solubilization were studied . The liquefied/solubilized products from coal by microorganisms were analysed . The liquid products analysed by IR titration and UV/visible spectrometry showed some alterations with regard to the original coal . Humic acids extracted from the liquefied lignite showed a reduction in the average molecular weight and a increase in the condensation index, probably due to depolymerization caused by microorganisms . The mechanisms implicated in coal biosolubilization by two fungal strains, M2 (Trichoderma sp.) and M4 (Penicillium sp.) were also studied . Extracellular peroxidase, esterase and phenoloxidase enzymes appear to be involved in coal solubilization. Appl Microbiol Biotechnol, 1999 Jul, 52(1), 25 - 40 Biotechnology and microbiology of coal degradation Fakoussa RM, Hofrichter M. For several years it has been known that fungi and bacteria can attack and even liquefy low rank coals . This review covers the progress in coal biotechnology and microbiology, mainly during the last decade, from describing the first effects to elucidating the mechanisms used by the microorganisms . More than one mechanism is responsible for microbial coal degradation/liquefaction: oxidative enzymes (peroxidases, laccases), hydrolytic enzymes (esterases), alkaline metabolites and natural chelators . Due to the heterogeneous structure of coal, which is described in one section, and for economic reasons the review focuses on the enzymatic depolymerization of brown coal . Approaches which seem not so promising are discussed (anaerobic, reductive pathways, chemical pretreatment) . Finally the possible applications and products in this field are summarized, as lignite with a worldwide production of about 940 million tons a year will continue to play an important economic role in the future. Mil Med, 1999 Aug, 164(8), 541 - 2 Environmental air sampling to detect biological warfare agents; Campbell J et al.; The rapid and unequivocal detection and identification of biological warfare agents is a major goal of military and civilian defense authorities . To identify agents of concern in an environmental sample, a reliable, region-specific characterization of the microorganisms found naturally at the sampling location is required . We have analyzed environmental air samples from Korea, Kuwait, and Bahrain by polymerase chain reaction and temporal temperature gradient electrophoresis and have produced genetic fingerprints of the natural microbial flora in these regions . Results are displayed as specific bar code patterns against which the unique patterns of potential biological warfare agents appearing in a sample can be quickly discriminated . Data are stored on compact disk, along with other laboratory analyses and relevant meteorological data, and are available to appropriate authorities and researchers. Arch Pathol Lab Med, 1999 Sep, 123(9), 835 - 7 Coinfection of visceral leishmaniasis and Mycobacterium in a patient with acquired immunodeficiency syndrome; Wang J et al.; We report a case of coinfection of visceral leishmaniasis and Mycobacterium avium-intracellulare in the same lesions in the small bowel and bone marrow of a 33-year-old man with acquired immunodeficiency syndrome who complained of abdominal pain and chronic diarrhea . The duodenal mucosa and bone marrow biopsy specimens showed numerous foamy macrophages packed with two forms of microorganisms that were identified histologically and ultrastructurally as Leishmania and Mycobacterium species . Visceral leishmaniasis is rarely suspected in patients residing in nonendemic countries including the United States . It should be included in the differential diagnosis for opportunistic infection in patients with acquired immunodeficiency syndrome . An appropriate travel history is important . To our knowledge, this is the first reported case showing coinfection of visceral leishmaniasis and Mycobacterium avium-intracelluulare in the same lesion in a patient with acquired immunodeficiency syndrome. Afr J Med Med Sci, 1996 Sep, 25(3), 255 - 9 Prognostic factors in childhood intra-abdominal sepsis; Adejuyighe O et al.; Over a 36-month period, prognostic factors in childhood intra-abdominal abscesses were evaluated in fifty-five Nigerian children (38 boys {69.1%} and 17 girls {30.9%} aged 2 months-15 years (mean 8.9, SD 4.3 years) . The mortality rate was 20% . The adverse prognostic factors were: a high fever, a positive blood culture, subphrenic location of abscesses, abscesses associated with a typhoid perforation, post-operative abscesses, presence of organ impairment, and multiple abscesses occurring together in the same patient . The age of the patient, duration of illness before hospitalization as well as the number and type of microorganisms cultured from the abscess did not have any influence on the prognosis in the children in this study. J Food Prot, 1999 Aug, 62(8), 845 - 9 Survival of enterohemorrhagic Escherichia coli O157:H7 in bovine feces applied to lettuce and the effectiveness of chlorinated water as a disinfectant; Beuchat LR; Bovine feces are a potential vehicle for transmitting enterohemorrhagic Escherichia coli O157:H7 to humans . A study was undertaken to determine survival characteristics of E . coli O157:H7 on iceberg lettuce using 0.1% peptone water and bovine feces as carriers for inocula . Four levels of inoculum, ranging from 10(0) to 10(5) CFU of E . coli O157:H7 per g of lettuce, were applied . Populations surviving on lettuce stored at 4 degrees C were monitored for up to 15 days . Regardless of the type of carrier, viable cells of E . coli O157:H7 were detected on lettuce after 15 days, even when the initial inoculum was 10(0) to 10(1) CFU/g . Spray treatments of lettuce with 200 ppm chlorine solution or deionized water were equally effective in killing or removing E . coli O157:H7 from lettuce . Holding lettuce for 5 min after spray treatment was not more effective in reducing populations than holding for 1 min before rinsing with water . Prevention of contamination of lettuce with bovine feces that may harbor E . coli O157:H7 as well as other infectious microorganisms is essential to minimizing the risk of illness . The development of sanitizers more efficacious than chlorine for the removal of pathogens from raw fruits and vegetable is needed. Braz J Med Biol Res, 1999 Jul, 32(7), 835 - 9 Searching for the role of protein phosphatases in eukaryotic microorganisms; da-Silva AM et al.; Preference for specific protein substrates together with differential sensitivity to activators and inhibitors has allowed classification of serine/threonine protein phosphatases (PPs) into four major types designated types 1, 2A, 2B and 2C (PP1, PP2A, PP2B and PP2C, respectively) . Comparison of sequences within their catalytic domains has indicated that PP1, PP2A and PP2B are members of the same gene family named PPP . On the other hand, the type 2C enzyme does not share sequence homology with the PPP members and thus represents another gene family, known as PPM . In this report we briefly summarize some of our studies about the role of serine/threonine phosphatases in growth and differentiation of three different eukaryotic models: Blastocladiella emersonii, Neurospora crassa and Dictyostelium discoideum . Our observations suggest that PP2C is the major phosphatase responsible for dephosphorylation of amidotransferase, an enzyme that controls cell wall synthesis during Blastocladiella emersonii zoospore germination . We also report the existence of a novel acid- and thermo-stable protein purified from Neurospora crassa mycelia, which specifically inhibits the PP1 activity of this fungus and mammals . Finally, we comment on our recent results demonstrating that Dictyostelium discoideum expresses a gene that codes for PP1, although this activity has never been demonstrated biochemically in this organism. Pharmacotherapy, 1999 Aug, 19(8), 995 - 1001 Phaeohyphomycosis from Exphiala jeanselmei with concomitant Nocardia asteroides infection in a renal transplant recipient: case report and review of the literature; Sartoris KE et al.; A 59-year-old black man who received a cadaveric renal transplant 15 months earlier developed subcutaneous nodules on his right upper extremity that were identified as phaeohyphomycosis caused by Exophiala jeanselmei . The man was admitted 4 weeks later with a swollen left arm and had Nocardia asteroides in this area and in the apex of his left lung . He was treated with surgical excision, and itraconazole, imipenem-cilastatin, and trimethoprim-sulfamethoxazole . With the potential presence of more than one microorganism in an immunocompromised patient, it is important to identify and differentiate them correctly to direct appropriate therapy. Ann Otol Rhinol Laryngol, 1999 Aug, 108(8), 762 - 8 Mucin gene expression in the rat middle ear: an improved method for RNA harvest; Lin J et al.; Mucins are heavily glycosylated proteins characterized by high molecular weight and heterogeneous structure . Mucin genes are expressed in a tissue- or epithelium-specific manner . Although mucins are known to be important structural components of the mucociliary transport system that protects epithelium against invading microorganisms, very little is known about mucin gene expression unique to the middle ear . This study demonstrated that middle ear messenger RNA specifically hybridized with rat MUC2 and human MUC2 (SMUC-41) complementary DNA probes . MUC3 and MUC5AC mucin genes, dominantly expressed in rodent intestine and trachea, were not detected in the rat middle ears in this study . The middle ear MUC2 messenger RNA harvested by lavage was characterized by a single transcript--unlike its counterpart in intestine and airways, which is characterized by polydispersity--suggestive of a better method for RNA analysis . It was concluded that rat middle ears possess a MUC2 mucin gene or homologue of human MUC2 (SMUC-41). Clin Infect Dis, 1999 May, 28(5), 1043 - 8 Conventional respiratory viruses recovered from immunocompromised patients: clinical considerations; Rabella N et al.; Lower respiratory tract infection is the most common complication in the immunocompromised patient . From January 1991 to December 1995, 785 consecutive patients with suspected respiratory tract infections were studied . One hundred ninety-nine viruses were isolated from 182 (23%) of 785 bronchoalveolar lavage fluid specimens . Cytomegalovirus was isolated from 131 patients, herpes simplex virus was recovered from 31, and conventional respiratory viruses (CRVs) were recovered from 36 . There were 9 influenza A viruses, 2 influenza B viruses, 7 parainfluenza viruses, 5 respiratory syncytial viruses, 5 adenoviruses, 6 enteroviruses, and 3 rhinoviruses . We identified 22 patients from whom a CRV was the only microorganism recovered; 13 patients developed pneumonia, 10 had acute respiratory failure, 5 required support with mechanical ventilation, and 5 (23%) died . In conclusion, CRVs are frequent causes of respiratory illnesses and are associated with high rates of morbidity and mortality among immunocompromised patients. Electrophoresis, 1999 Jul, 20(10), 2100 - 10 Proteomics in human disease: cancer, heart and infectious diseases; Jungblut PR et al.; In recent years, genomics has increased the understanding of many diseases . Proteomics is a rapidly growing research area that encompasses both genetic and environmental factors . The protein composition represents the functional status of a biological compartment . The five approaches presented here resulted in the detection of disease-associated proteins . Calgranulin B was upregulated in colorectal cancer, and hepatoma-derived aldose reductase-like protein was reexpressed in a rat model during hepatocarcinogenesis . In these two investigations, attention was focused on one protein, obviously differing in amount, directly after two-dimensional electrophoresis (2-DE) . Additional methods, such as enzyme activity measurements and immunohistochemistry, confirmed the disease association of the two candidates resulting from 2-DE subtractive analysis . The following three investigations take advantage of the holistic potential of the 2-DE approach . The comparison of 2-DE patterns from dilated cardiomyopathy patients with those of controls revealed 25 statistically significant intensity differences, from which 12 were identified by amino acid analysis, Edman degradation or matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) . A human myocardial 2-DE database was constructed, containing 3300 protein spots and 150 identified protein species . The number of identified proteins was limited by the capacity of our group, rather than by the principle of feasibility . Another field where proteomics proves to be a valuable tool in identifying proteins of importance for diagnosis is proteome analysis of pathogenic microorganisms such as Borrelia burgdorferi (Lyme disease) and Toxoplasma gondii (toxoplasmosis) . Sera from patients with early or late symptoms of Lyme borreliosis contained antibodies of various classes against about 80 antigens each, containing the already described antigens OspA, B and C, flagellin, p83/100, and p39 . Similarly, antibody reactivity to seven different marker antigens of T . gondii allowed differentiation between acute and latent toxoplasmosis, an important diagnostic tool in both pregnancy and immunosuppressed patients. Electrophoresis, 1999 Jul, 20(10), 2094 - 8 An assessment of protein profiles from the marine oligotrophic ultramicrobacterium, Sphingomonas sp . strain RB2256; Fegatella F et al.; The protein expression profile of a novel marine oligotrophic ultramicrobacterium, Sphingomonas sp . strain RB2256, was investigated by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) . Analytical reference maps were generated from mid-log phase batches and steady-state chemostat cultures with pH 4-8 immobilised pH gradients (IPGs) followed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis . The resolved proteins were detected by two different methods: radioactive labeling and silver staining . Protein profiles generated from analytical 2-D PAGE gels were compared and differential analysis was performed using Melanie II software . Both methods (radioactive labeling and silver staining) resulted in reproducible, high resolution gels (up to 1600 protein spots) . This approach is proving to be a powerful tool for investigating the molecular basis of the unique physiology of this model oligotrophic microorganism. Scand J Work Environ Health, 1999 Jun, 25(3), 238 - 45 Exposure-response relationship between gastrointestinal problems among waste collectors and bioaerosol exposure; Ivens UI et al.; OBJECTIVES: A high frequency of gastrointestinal problems has previously been reported for waste collectors . The present study relates the gastrointestinal problems to measurements of the bioaerosols that waste collectors are exposed to during work . METHODS: A job-exposure matrix was constructed from a combination of questionnaire data and field measurements . The questionnaire data were collected from 1747 male waste collectors and a comparison group of 1111 male municipal workers . Moreover a total of 189 full-shift personal samples was collected . The samples were used for characterizing the bioaerosol exposure described by viable fungi, total count of fungal spores, microorganisms, and endotoxins . RESULTS: In a multivariate analysis, high exposure to endotoxins was associated with nausea {prevalence proportion ratio (PPR) 1.60}, and the risk of reporting nausea decreased with decreasing exposure so that workers with low exposure had the fewest reports (PPR 1.39) in the comparison with the unexposed group . High exposure to endotoxins was also associated with reports of diarrhea (PPR 5.60), and the risk of reporting diarrhea decreased with decreasing exposure so that the workers with low exposure had the fewest reports (PPR 3.02) . The same pattern existed for exposure to fungi, for which high exposure resulted in the most reports (PPR = 4.59), and for diarrhea, for which low exposure resulted in the fewest reports (PPR = 3.15) . CONCLUSIONS: An exposure-response relationship was found between nausea and endotoxin exposure and between diarrhea and exposure to both endotoxins and viable fungi. Infect Dis Obstet Gynecol, 1999, 7(4), 195 - 8 Increased susceptibility to Helicobacter pylori infection in pregnancy; Lanciers S et al.; OBJECTIVE: Helicobacter pylori plays a major role in abdominal symptoms and gastroduodenal pathology, including gastric cancer . Pregnancy has been associated with changes in both humoral and cell-mediated immunity . These changes include alterations in the various classes of antibodies during different gestational periods . It has been previously suggested that these alterations may expose pregnant women to an increased risk of infection with this microorganism . METHODS: To further investigate this hypothesis, we assayed sera from 229 asymptomatic pregnant women for the presence of H.-pylori-specific immunoglobulin G (IgG) and immunoglobulin M (IgM) antibodies by means of a commercially available serum ELISA test (Malakit, Biolab, Belgium) . Both tests were previously validated in large series of H.-pylori-positive and -negative subjects . While the presence of H.-pylori-specific IgG antibodies is only a marker for a "chronic" infection with this bacterium and therefore no indicator of the time of acquisition of the infection, specific IgM antibodies are a more specific marker for a recently acquired infection with H . pylori . Results were compared with those previously obtained in asymptomatic, healthy, nonpregnant individuals . RESULTS: One hundred twenty of 229 women (52.4%) and 55/118 nonpregnant subjects (46.6%) were seropositive for H.-pylori-specific IgG antibodies (P > 0.3) . Out of these 120 IgG-antibody-positive women, 36 (30%) were positive for H.-pylori-specific IgM antibodies, as were 25/109 (22.9%) in the IgG-antibody-negative group (P > 0.3) . Overall, 61/229 (26.6%) of the pregnant women had recently been infected with H . pylori, compared with 11% of the healthy, nonpregnant population (P > 0.01) . CONCLUSIONS: Our observations confirm the possibility of an increased susceptibility to H . pylori infection in pregnancy . Additional studies are necessary to further understand the immune response to H . pylori in pregnancy. Prev Vet Med, 1999 Jul 20, 41(2-3), 119 - 33 Risk factors for the between-herd spread of Mycobacterium bovis in Canadian cattle and cervids between 1985 and 1994; Munroe FA et al.; Microorganisms of the genus Mycobacterium cause tuberculosis in many animal species including humans . Generally, Mycobacterium bovis (M . bovis) infects cattle and cervids, but it has the potential to infect virtually all species of mammals . This study examined and analysed the data from the nine outbreaks of tuberculosis in Canadian cattle and cervids from 1985 to 1994 . For the purposes of this study, a positive herd was one with at least one culture-positive animal . A reactor herd had at least one animal which was positive or suspicious on a mid-cervical, comparative cervical, or gross or histopathologic test for tuberculosis . Herd classification was either reactor/positive or negative . Data for the study were collected from the outbreak records in the Regional or District offices of Agriculture and Agri-Food Canada . Logistic regression was used to study spread of tuberculosis between herds . Two risk factors were identified: increasing herd size; and, the reason why a herd was investigated as part of the outbreak . This latter factor was interpreted as a surrogate measure for the nature of contact between the study herd and other potentially infected herds in the outbreak . Increasing herd size was associated with an increased risk of being positive for tuberculosis with herds of 16-35, 36-80, and >80 animals having odds ratios of 2.9, 5.8, and 9.3, respectively, when compared to a herd size of <16 animals (p < 0.001) . When compared to perimeter testing (i.e . testing herds within a specified radius of an infected herd), all other reasons for investigation had higher odds ratios (p < 0.001) . These odds ratios were 57.8 for traceout herds (i.e . herds which had purchased animal(s) from a reactor/positive herd), 31.8 for herds with pasture or fence-line contact with a reactor/positive herd, and 14.9 for traceback herds (i.e . herds which had been a source of animals for reactor/positive herd(s)). J Math Biol, 1999 Aug, 39(2), 172 - 92 Effective drift velocities and effective diffusivities of swimming microorganisms in external flows; Yannacopoulos AN et al.; In this note we calculate the effective drift velocities and the effective diffusivities of swimming microorganisms in an external flow field . It is shown that if the ambient velocity field is incompressible then the effects of reorientation of the cells can under certain circumstances enhance the effective drift velocity along the preferred direction. Intensive Care Med, 1999 Aug, 25(8), 865 - 8 Severe non-infectious circulatory shock related to hypopituitarism; Hazouard E et al.; The authors report a case of refractory non-infectious circulatory shock with catecholamine and massive fluid loading-resistant features related to hypopituitarism . A 76-year-old man was admitted for shock after suffering from gastroenteritis for 3 days . He was pale and had sparse axillary and pubic hair and small testes . Right catheterization showed shock with low preload pressure and a low oxygen extraction ratio relevant for septic shock . Ultrasound tomography revealed a distended gallbladder due to a stone without peritoneal effusion . A non-inflammatory hydrops of the gallbladder was removed surgically . No microorganism was isolated . Cerebral computed tomography (CT) scan showed a pituitary mass . In the post-surgical period the shock became uncontrollable . Cortisol replacement therapy was instituted and clinical and hemodynamic improvement occurred after 2 h . Hormonal screening on admission before catecholamine administration showed a major decrease in all the hypothalamic-pituitary hormone concentrations . The patient died on day 15 with multiple organ failure . Hypopituitarism, probably owing to pituitary adenoma, was the only disease identified in this case . Hormone replacement therapy dramatically improved the clinical and hemodynamic status, although the role of an abdominal sepsis could not be eliminated . Arguments that pituitary hormone deficiency might increase the hemodynamic consequences of adrenal deficiency are discussed. Thyroid, 1999 Jul, 9(7), 643 - 6 The immunology of pregnancy; Weetman AP; Pregnancy is an immunological balancing act in which the mother's immune system has to remain tolerant of paternal major histocompatibility (MHC) antigens and yet maintain normal immune competence for defense against microorganisms . The placenta separates fetal and maternal blood and lymphatic systems and it is fetal trophoblast that plays the major role in evading recognition by the maternal immune system . Trophoblast cells fail to express MHC class I or class II molecules and the extravillous cytotrophoblast cells strongly express the nonclassic MHC gene encoding HLA-G, which may downregulate natural killer (NK) cell function . In addition, the trophoblast expresses Fas ligand, thereby conferring immune privilege: maternal immune cells expressing Fas will undergo apoptosis at the placenta/decidua interface . A third protective mechanism exploited by the trophoblast is the expression of the complement regulatory proteins CD46, CD55, and CD59 . Uterine decidual and placental cells produce a huge array of cytokines which, in part, contribute to the deviation of the immune response from Th1 to Th2 . This may leave the mother more open to infection whose control is Th1-dependent, but increased production of Th1 cytokines has been linked to spontaneous abortion and small-for-dates babies . This bias in cytokines and hormonally mediated effects on the thymus and on B cells may also contribute to the suppression of autoimmune responses and changes in circulating and local T-cell subsets in pregnancy. Appl Occup Environ Hyg, 1999 May, 14(5), 317 - 28 A multi-zone model evaluation of the efficacy of upper-room air ultraviolet germicidal irradiation; Nicas M et al.; Engineering controls can be used to reduce the spread of airborne infectious disease, particularly tuberculosis (TB), in high-risk settings . This article evaluates published data on the efficacy of upper-room air ultraviolet germicidal irradiation (UVGI) . A three-zone representation of a TB patient room equipped with a germicidal UV lamp is developed . The lamp irradiates the upper-room zone and inactivates airborne mycobacteria; the unirradiated lower-room zone also contains a near-field zone surrounding the TB patient . Infectious particles are generated in the near-field zone and transported throughout the room by air flow between zones . Each zone is independently well-mixed; the whole room, however, is not well-mixed . The three-zone model is applied to a previously published study of UVGI against airborne mycobacteria in a test room . Based on the estimated slopes of the semi-log concentration decay curves for viable mycobacteria, and on the assumption that the test room was essentially well-mixed, the published study reported that UVGI provided 10 to 25 equivalent air changes per hour . However, when the same decay curve slopes are interpreted in the context of the three-zone model, UVGI is seen to be far less effective in reducing exposure intensity near the TB patient . Near-field exposure intensity is relevant because health care workers are usually in close proximity to the TB patients they attend . In general, the interpretation of concentration decay data depends on the specific model of room air mixing that is assumed appropriate . It is recommended that tests of the efficacy of UVGI and other control devices against airborne microorganisms be based on steady-state concentration measurements rather than concentration decay measurements, because the former measurements do not require inferences based on a particular model. J Microbiol Methods, 1999 Aug, 37(2), 155 - 64 The potential of in situ hybridization and an immunogold assay to identify Legionella associations with other microorganisms; Desai R et al.; Based on in vitro studies, bacteria in the genus Legionella are believed to multiply within protozoa such as amoebae in aquatic environments . Current methods used for detection of Legionella species, however, are not designed to show this relationship . Thus the natural intimate association of Legionella with other microorganisms remains to be clearly documented and the extent to which protozoa might be infected with Legionella species remains undefined . In this report we describe methods based on the use of Legionella specific reagents that would prove useful in describing its associations with other microorganisms . An immunogold and in situ hybridization technique have the potential to demonstrate the natural occurrence of Legionella species in free-living amoebae . In preliminary observations, however, bacteria reactive with Legionella specific reagents were often not intimately associated with amoebae . Bacteria occurred as free single cells, as cell aggregates, in proximity to other cells and debris, and only occasionally in close proximity to amoebae . Although some Legionella species replicate within amoebae, these preliminary observations suggest the bacteria may be encountered most frequently as extracellular microorganisms, either free-floating or in association with other structures or microorganisms . The future use of these techniques will aid in the elucidation of any naturally occurring relationships between Legionella species and other microorganisms. J Periodontal Res, 1999 May, 34(4), 203 - 13 Antigen activation of THP-1 human monocytic cells after stimulation with lipopolysaccharide from oral microorganisms and granulocyte-macrophage colony-stimulating factor; Baqui AA et al.; A human THP-1 monocyte cell line culture system has been utilized to evaluate the morphological changes in THP-1 cells and to measure expression of activation antigens (CD-11b, CD-11c, CD-14, CD-35, CD-68, CD-71 and HLA-DR) as evidence of maturation of THP-1 cells in response to stimulation by lipopolysaccharide (LPS) from the oral microorganisms, Fusobacterium nucleatum and Porphyromonas gingivalis, and granulocyte-macrophage colony-stimulating factor . THP-1 cells were stimulated with LPS (1 microgram/ml) of P . gingivalis or F . nucleatum for different time periods (1, 2, 4 and 7 d) . Detection of different activation antigens on THP-1 cells was performed by indirect immunohistochemical staining followed by light microscopy . Confirmational studies were performed in parallel using indirect immunofluorescence and immunogold electron microscopy for detection of the corresponding activation antigens . Expression of different activation antigens by resting THP-1 cells revealed HLA-DR to be on 3% of the cells; CD-11b, 9%; CD-11c, 8%; CD-14, 22%; CD-35, 9% and CD-68, 7% . The CD-71 activation antigen was not expressed in untreated THP-1 cells . LPS stimulation increased expression of all activation antigens . A significant (p < 0.05) increase in expression of CD-11b, CD-11c, CD-14, CD-35, CD-68 and CD-71 was observed when GM-CSF (50 IU/ml) was supplemented during the treatment of THP-1 cells with LPS of F . nucleatum or P . gingivalis . Activation and differentiation of THP-1 cells by LPS from oral microorganisms in the presence of GM-CSF supports a role for human macrophages in acute and chronic periodontal diseases and may explain the clinically observable periodontal exacerbations in some patients after GM-CSF therapy. Am J Physiol, 1999 Aug, 277(2 Pt 1), C271 - 9 Human colonic subepithelial myofibroblasts modulate transepithelial resistance and secretory response; Beltinger J et al.; The epithelium of the gastrointestinal tract transports ions and water but excludes luminal microorganisms and toxic molecules . The factors regulating these important functions are not fully understood . Intestinal myofibroblasts lie subjacent to the basement membrane, at the basal surface of epithelial cells . We recently showed that primary cultures of adult human colonic subepithelial myofibroblasts express cyclooxygenase (COX)-1 and COX-2 enzymes and release bioactive transforming growth factor-beta (TGF-beta) . In this study we have investigated the role of normal human colonic subepithelial myofibroblasts in the regulation of transepithelial resistance and secretory response in HCA-7 and T84 colonic epithelial cell lines . Cocultures of epithelial cells-myofibroblasts and medium conditioned by myofibroblasts enhanced transepithelial resistance and delayed mannitol flux . A panspecific antibody to TGF-beta (but not piroxicam) antagonized this effect . In HCA-7 cells, myofibroblasts downregulated secretagogue-induced change in short-circuit current, and this effect was reversed by pretreatment of myofibroblasts with piroxicam . In contrast to HCA-7 cells, myofibroblasts upregulated the agonist-induced secretory response in T84 cells . This study shows that intestinal subepithelial myofibroblasts enhance barrier function and modulate electrogenic chloride secretion in epithelial cells . The enhancement of barrier function was mediated by TGF-beta . In contrast, the modulation of agonist-induced change in short-circuit current was mediated by cyclooxygenase products . These findings suggest that colonic myofibroblasts regulate important functions of epithelial cells via distinct secretory products. Nature, 1999 Jul 29, 400(6743), 476 - 80 Structure of cytochrome c nitrite reductase; Einsle O et al.; The enzyme cytochrome c nitrite reductase catalyses the six-electron reduction of nitrite to ammonia as one of the key steps in the biological nitrogen cycle, where it participates in the anaerobic energy metabolism of dissimilatory nitrate ammonification . Here we report on the crystal structure of this enzyme from the microorganism Sulfurospirillum deleyianum, which we solved by multiwavelength anomalous dispersion methods . We propose a reaction scheme for the transformation of nitrite based on structural and spectroscopic information . Cytochrome c nitrite reductase is a functional dimer, with 10 close-packed haem groups of type c and an unusual lysine-coordinated high-spin haem at the active site . By comparing the haem arrangement of this nitrite reductase with that of other multihaem cytochromes, we have been able to identify a family of proteins in which the orientation of haem groups is conserved whereas structure and function are not. J Periodontol, 1999 Jul, 70(7), 761 - 71 Longitudinal effect of non-surgical treatment and systemic metronidazole for 1 week in smokers and non-smokers with refractory periodontitis: a 5-year study; Soder B et al.; BACKGROUND: Periodontitis consists of a mixture of diseases, most of which respond favorably to traditional mechanical therapy . It is now recognized that advanced periodontitis does not always respond to conventional management with scaling, periodontal surgery, and oral hygiene measures . However, various types of antibiotics given systemically or locally improve the success rate of periodontal therapy . In short-term studies, it has been shown that metronidazole, when systemically administered after debridement, resulted in treatment benefits including less need for surgical intervention . METHODS: In this double-blind study, we evaluated periodontal treatment involving initial non-surgical treatment, systemic administration of metronidazole for 1 week, and then follow-ups for scaling and root planing every 6 months, for 5 years . The study population consisted of 64 subjects (37 smokers and 27 non-smokers), mean age 36.3 (+/-3.0 SD) years, with severe periodontal disease . After initial scaling and root planing, patients were randomly assigned to the intervention or placebo groups: 400 mg metronidazole or a placebo administered at 8-hour intervals for 1 week . The participants underwent an extensive clinical periodontal examination . Gingival crevicular fluid (GCF) was analyzed for spirochetes and granulocytes . Samples were cultured for Actinobacillus actinomycetemcomitans (A.a.), Porphyromonas gingivalis (Pg.), and Prevotella intermedia (Pi.) . RESULTS: The number of patients infected with A.a., Pg., Pi., and spirochetes decreased during the study . Most patients who harbored spirochetes at the end of the study had these microorganisms at the beginning . Smokers responded less favorably to periodontal therapy than non-smokers . Non-smoking patients who required only non-surgical therapy in the intervention group showed statistically significant improvement in the clinical parameters after 5 years . Patients with complete healing, defined as the absence of inflamed sites > or =5 mm, after 5 years were found only in the intervention group . The patients considered healthy after 5 years were the same patients found to be healthy after 6 months . CONCLUSIONS: Decisive factors in the sustained long-term improvement of patients who respond satisfactorily to treatment are probably initial scaling and root planing; a brief course of metronidazole; and regular follow-up examinations at 6-month intervals for oral hygiene and scaling and root planing. Am J Clin Pathol, 1999 Aug, 112(2), 204 - 10 Epithelial cell atypia in bronchoalveolar lavage specimens from lung transplant recipients; Ohori NP; In lung transplant recipients, bronchoalveolar lavage (BAL) is mainly performed to detect infectious agents . However, in addition to microorganisms, epithelial cell atypia may be identified, and determination of its significance is necessary . Specimens obtained at BAL in lung and heart-lung transplant recipients (LTRs) between 1991 and 1998 were examined for the presence of significant cytologic atypia in epithelial cells . Ten cases in 9 patients were identified, and these composed the core of our study . These transplant BAL specimens were compared with 4 BAL specimens with carcinoma from non-transplant patients (NTPs) . Fourteen cytologic parameters were evaluated, and clinical and biopsy correlation was made in each case . Significant overlap in cytologic features, including background cellularity, number of atypical cell clusters, number of cells in each cluster, size of cell clusters, contour of clusters, 3-dimensionality, tenacious intercytoplasmic connections, multinucleation, nuclear size, nuclear/cytoplasmic ratio, nuclear membrane irregularity, chromatin pattern, intranuclear inclusions, and nucleolar characteristics, was observed between atypical LTR cases and NTP carcinoma cases . Clinically, all LTR cases derived from nonneoplastic conditions including harvest injury (diffuse alveolar damage), acute cellular rejection, and infections . Our study results show that evaluation of cytologic features alone does not permit differentiation of atypical cells found in nonneoplastic conditions from those in malignant conditions . Clinical and histologic correlation and awareness of the range of atypia seen in posttransplant syndromes is important in correct interpretation of these cases. Electrophoresis, 1999 Jun, 20(8), 1722 - 8 DNA preservation: a microsatellite-DNA study on ancient skeletal remains; Burger J et al.; To determine the effect of environmental factors on the preservation of DNA, archeological teeth of approximately similar age but greatly differing site milieu were examined for DNA content . The complex relational system of locational milieu of the samples was reduced to its essential and, at the same time, easily measurable factors . These are temperature, humidity, pH value, the geochemical properties of the soil, the amount of postmortal organic substances and the general degree of microbial infestation in the respective soil . The relative DNA content in the samples was established by determining the rate of successful polymerase chain reaction (PCR) amplifications . Differences in quantity and quality of the results are attributed to the respective prevailing environmental factor or to the respective storage conditions . Dryness, low temperature and absence of microorganisms favors the preservation of DNA . The bioapatite of bones and teeth, like the DNA, are preserved under neutral or slightly alkaline conditions . Brief storage at room temperature does not affect the amount of amplifiable DNA but does affect the reproducibility of the results . Long storage outside a lab freezer reduces the amount and the reproducibility of DNA amplifications in ancient specimens. Gene, 1999 Aug 5, 236(1), 149 - 57 Genetic characterization and evolutionary implications of a chromosomally encoded naphthalene-degradation upper pathway from Pseudomonas stutzeri AN10; Bosch R et al.; Pseudomonas stutzeri strain AN10 is a naphthalene-degrading strain whose dissimilatory genes are chromosomally encoded . We sequenced a total of 11514bp including the entire naphthalene-degradation upper pathway (nah) of P . stutzeri AN10 . Nine open reading frames, nahAaAbAcAdBFCED, encoding the enzymes for the degradation of naphthalene to salicylate, were identified . The nah genes of P . stutzeri AN10 have been compared with genes encoding isofunctional proteins from other Pseudomonas naphthalene-degradation upper pathways . The implications of the sequence homologies to the evolution of aromatic catabolic pathways are discussed . Our findings indicate that this entire catabolic module of P . stutzeri AN10 was recruited from other microorganisms and a short period of time has elapsed after its incorporation within the P . stutzeri AN10 genome . Comparisons also suggest the coexistence of two entire nah upper pathways in a host strain, and further recombination between them . These events could accelerate the evolution of modern catabolic pathways. Development, 1999 Sep, 126(17), 3735 - 45 Ontogeny and behaviour of early macrophages in the zebrafish embryo; Herbomel P et al.; In the zebrafish embryo, the only known site of hemopoieisis is an intra-embryonic blood island at the junction between trunk and tail that gives rise to erythroid cells . Using video-enhanced differential interference contrast microscopy, as well as in-situ hybridization for the expression of two new hemopoietic marker genes, draculin and leucocyte-specific plastin, we show that macrophages appear in the embryo at least as early as erythroid cells, but originate from ventro-lateral mesoderm situated at the other end of the embryo, just anterior to the cardiac field . These macrophage precursors migrate to the yolksac, and differentiate . From the yolksac, many invade the mesenchyme of the head, while others join the blood circulation . Apart from phagocytosing apoptotic corpses, these macrophages were observed to engulf and destroy large amounts of bacteria injected intravenously; the macrophages also sensed the presence of bacteria injected into body cavities that are isolated from the blood, migrated into these cavities and eradicated the microorganisms . Moreover, we observed that although only a fraction of the macrophage population goes to the site of infection, the entire population acquires an activated behaviour, similar to that of activated macrophages in mammals . Our results support the notion that in vertebrate embryos, macrophages endowed with proliferative capacity arise early from the hemopoietic lineage through a non-classical, rapid differentiation pathway, which bypasses the monocytic series that is well-documented in adult hemopoietic organs. Am J Infect Control, 1999 Aug, 27(4), 315 - 9 Levels of naturally occurring microorganisms on surgical instruments after clinical use and after washing; Chu NS et al.; Surgical instruments exposed to sterile body sites should be contaminated with relatively low levels of microbial contamination or bioburden; however, few studies in the literature have determined the quantitative level and types of contamination . A study was conducted at 2 clinical sites to determine the level of microbial contamination of surgical instruments after clinical use and after washing . Quantitative assays showed that bioburden levels were in the range of 0 to 4415 colony forming units per instrument after clinical use, and 88% of the instruments had bioburden levels lower than 1000 . As expected, a reduction in counts occurred after washing; however, in some cases, higher counts were found on the instruments after the washing process . Although the washing procedure is effective in reducing the microbial levels deposited on the surgical instruments during use, a recontamination process occurs that results in increased counts after washing . The low bioburden level after washing consists of predominantly vegetative microorganisms that present a relatively low challenge to sterilization and disinfection systems. J Neurosurg, 1999 Aug, 91(2), 330 - 4 Idiopathic pinealitis . Case report; Nikas DC et al.; This 63-year-old man presented with complaints of "having a feeling of falling backward" over a 3-month period . Results of his general physical examination, laboratory studies, and neurological examination were unremarkable . A magnetic resonance image revealed a 1.8 x 1.4 x 1.2-cm enhancing mass in the posterior third ventricle just above the corpora quadrigemina . The pineal gland was found to be diffusely enlarged at operation and separable from the posterior thalamus and was totally resected . The patient had an uneventful postoperative course but continues to be somewhat confused . The lesion consisted of a remarkable chronic inflammatory cell infiltrate permeating the pineal lobules and was composed of T and B lymphocytes, macrophages, eosinophils, and mast cells . Immunoperoxidase studies did not demonstrate Langerhans cells, and a search for microorganisms was unrevealing . There was no evidence of neoplasia; results of immunostaining for germ cell markers and other tumor-associated antigens were negative. Rev Clin Esp, 1999 Jun, 199(6), 356 - 61 {A multicenter study on fungemia caused by yeasts in Spain (April-June, 1997) . A Work Group to Study Fungemia)}; Rodriguez-Tudela JL et al.; Yeast fungemia has increased markedly in the last few years . Currently, Candida spp . is one of the microorganisms recovered most frequently from blood cultures . To better know the prevalence of yeast fungemia in our country we conducted a cross-sectional study with yeasts recovered from blood cultures obtained during a 3-month period from 39 hospitals in the Spanish public health hospital net . A total of 153 yeasts were recovered (59 were C . albicans, 53 C . parapsilosis, and 41 other species of yeasts) . The percentage of males was 64.1 . Mean age was 50.8 years . Factors associated with the appearance of fungemia were previous antibiotic therapy (83.7%), venous catheter (66%), previous bacterial infections (47.7%), and parenteral nutrition (40.5%) . The recovery of C . albicans was independently associated with the admission to ICU, PDA and presence of urological/renal disease; C . parapsilosis was independently associated with the presence of hematological disease . The susceptibility studies of these strains recovered from blood cultures showed that in vitro resistance to amphotericin B and 5-fluorocytosine are practically nonexistent and that resistance to azole compounds is low . Multicentric epidemiologic studies are still necessary in this field of Microbiology. Obstet Gynecol, 1999 Aug, 94(2), 279 - 83 Apoptosis in placentas from human T-lymphotropic virus type I-seropositive pregnant women: a possible defense mechanism against transmission from mother to fetus; Fujino T et al.; OBJECTIVE: The mechanism by which the placenta serves as the barrier against mother-to-fetus transmission of microorganisms remains to be elucidated . Programmed cell death, apoptosis, is considered a cellular defense mechanism against infection . The hypothesis of this study is that apoptosis of human T-lymphotropic virus type I (HTLV-I)-infected placental villous cells is involved in the defense mechanism against mother-to-fetus transmission of HTLV-I . METHODS: Apoptosis was compared in term placentas from eight HTLV-I-seropositive pregnant women and eight HTLV-I-seronegative pregnant women by the terminal deoxynucleotidyl transferase-mediated deoxyuridine nick end-labeling method . In addition, an in vitro cocultivation with an HTLV-I-infected lymphocyte cell line (MT-2 cells) was performed to examine whether placental villous cells were infected with HTLV-I and apoptosis was induced . RESULTS: The incidence of apoptosis-positive cells (nuclei) in placentas from the HTLV-I-seropositive pregnant women was higher than in the HTLV-I-seronegative pregnant women (P < .02) . Cocultivation with MT-2 cells showed that trophoblast cells were able to be infected with HTLV-I and that apoptosis was induced in the placental villous cells . CONCLUSION: HTLV-I infection induces apoptosis in the placenta . We speculate that apoptosis may be involved in the defense mechanism of the placenta against mother-to-fetus transmission of HTLV-I. Arch Med Res, 1999 May-Jun, 30(3), 171 - 8 Secretion antigens of Mycobacterium tuberculosis: a comparison between a reference strain and seven wild isolates; Rojas-Espinosa O et al.; BACKGROUND: This study was carried out with the aim of detecting possible differences between proteins secreted by fresh wild isolates of Mycobacterium tuberculosis and from a reference strain of this microorganism, H37Rv TMCC 102 . MATERIALS AND METHODS: This reference strain of M . tuberculosis has been in our laboratory for over 10 years, where it has been maintained by serial subcultures in PBY and Lowenstein-Jensen media . Patterns of protein secretion and recognition by sera derived from both tuberculosis patients and normal individuals were analyzed by electrophoresis and Western blotting . RESULTS: No major qualitative differences were observed among the several strains studied with respect to protein patterns or recognition of these proteins by test sera . Normal sera were found to react with almost all antigens recognized by tuberculosis sera, but with less intensity . However, a small protein of 14.5 kDa, secreted by both the wild and reference strains of M . tuberculosis, was recognized by 32 of the 40 tuberculous patient sera tested (80%), and was not recognized by any of the 40 serum samples derived from healthy individuals . CONCLUSIONS: This small protein seems to be a potentially important antigen for the serological diagnosis of tuberculosis and/or for use in the follow-up of patients who received treatment. Appl Environ Microbiol, 1999 Aug, 65(8), 3641 - 50 Isolation and characterization of Methanomethylovorans hollandica gen . nov., sp . nov., isolated from freshwater sediment, a methylotrophic methanogen able to grow on dimethyl sulfide and methanethiol; Lomans BP et al.; A newly isolated methanogen, strain DMS1(T), is the first obligately anaerobic archaeon which was directly enriched and isolated from a freshwater sediment in defined minimal medium containing dimethyl sulfide (DMS) as the sole carbon and energy source . The use of a chemostat with a continuous DMS-containing gas stream as a method of enrichment, followed by cultivation in deep agar tubes, resulted in a pure culture . Since the only substrates utilized by strain DMS1(T) are methanol, methylamines, methanethiol (MT), and DMS, this organism is considered an obligately methylotrophic methanogen like most other DMS-degrading methanogens . Strain DMS1(T) differs from all other DMS-degrading methanogens, since it was isolated from a freshwater pond and requires NaCl concentrations (0 to 0.04 M) typical of the NaCl concentrations required by freshwater microorganisms for growth . DMS was degraded effectively only in a chemostat culture in the presence of low hydrogen sulfide and MT concentrations . Addition of MT or sulfide to the chemostat significantly decreased degradation of DMS . Transient accumulation of DMS in MT-amended cultures indicated that transfer of the first methyl group during DMS degradation is a reversible process . On the basis of its low level of homology with the most closely related methanogen, Methanococcoides burtonii (94.5%), its position on the phylogenetic tree, its morphology (which is different from that of members of the genera Methanolobus, Methanococcoides, and Methanohalophilus), and its salt tolerance and optimum (which are characteristic of freshwater bacteria), we propose that strain DMS1(T) is a representative of a novel genus . This isolate was named Methanomethylovorans hollandica . Analysis of DMS-amended sediment slurries with a fluorescence microscope revealed the presence of methanogens which were morphologically identical to M . hollandica, as described in this study . Considering its physiological properties, M . hollandica DMS1(T) is probably responsible for degradation of MT and DMS in freshwater sediments in situ . Due to the reversibility of the DMS conversion, methanogens like strain DMS1(T) can also be involved in the formation of DMS through methylation of MT . This phenomenon, which previously has been shown to occur in sediment slurries of freshwater origin, might affect the steady-state concentrations and, consequently, the total flux of DMS and MT in these systems. Appl Environ Microbiol, 1999 Aug, 65(8), 3398 - 400 Culture-independent identification of microorganisms that respond to specified stimuli; Borneman J; A new approach that permits culture-independent identification of microorganisms that respond to specified stimuli was developed . This approach was illustrated by examination of microorganisms that grew in response to various nutrient supplements added to soil . A thymidine nucleotide analog, bromodeoxyuridine (BrdU), and supplements were added to soil and incubated for 3 days . DNA was extracted from the soil, and the newly synthesized DNA was isolated by immunocapture of the BrdU-labeled DNA . The unique perspective this approach offers was demonstrated by comparing the microbial community structures obtained from total soil DNA and the BrdU-labeled fraction in an rRNA gene (rDNA) analysis . The traditional total DNA analysis revealed no notable differences between the treatments, whereas the BrdU-labeled DNA showed significantly different banding patterns between the nutrient supplement treatments and compared with total DNA banding patterns . PCR primers were developed to specifically amplify the intergenic region of an rDNA sequence unique to the BrdU analysis of a phosphate supplement treatment . Amplification of DNA from all treatments using these primers showed that it was unique to the phosphate treatment and that it was present in both the total DNA and BrdU-labeled DNA fractions . This result demonstrates the promise of this new strategy, because it was able to permit identification of a sequence from a phosphate-responsive organism that was not discernable in the traditional total DNA community structure analysis. Biochem Cell Biol, 1999, 77(1), 25 - 31 Expression of a mammalian Na+/H+ antiporter in Saccharomyces cerevisiae; Montero-Lomeli M et al.; The basolateral Na+/H+ antiporter (NHE) from LLC-PK1 cells was expressed in Saccharomyces cerevisiae . Two different strategies were tested for expression . In the first, we used a yeast strain that contains a temperature-sensitive mutation in the SEC-6 gene, whose product is required for the fusion of secretory vesicles with the plasma membrane . This strain was transformed with a vector containing the coding region of the NHE1 isoform under control of a heat shock (HS) promoter (pYNHE1-HS) . In the second strategy, we replaced the heat shock promoter from pYNHE1-HS with a galactose (GAL) promoter (pYNHEI-GAL) and transformed wild-type yeast . In both cases, Northern blots demonstrated a transcript that hybridized against a probe containing the membrane region of the exchanger . When an antibody against the last 40 amino acids of the carboxy-terminus of NHE1 was used for immunoblots, a protein with a Mr of 73000 was seen in total membranes from both yeast transformants . Subcellular fractionation revealed that NHE1 was expressed in the endoplasmic reticulum . In the case of the pYNHEI-GAL transformant, the 100000 x g membrane pellet was reconstituted in phosphatidylcholine liposomes, and ethylisopropylamiloride-sensitive Na+/H+ exchange was observed . These results have paved the way for expression of the Na+/H+ exchanger in a genetically well-known microorganism. Acta Paediatr, 1999 May, 88(5), 521 - 7 Interactions of infectious symptoms and modifiable risk factors in sudden infant death syndrome . The Nordic Epidemiological SIDS study; Helweg-Larsen K et al.; The aim of the study was to investigate the effect of infection on sudden infant death syndrome (SIDS) and to analyse whether modifiable risk factors of SIDS, prone sleeping, covered head and smoking act as effect modifiers . In a consecutive multicentre case-control study of SIDS in Denmark, Norway and Sweden, questionnaires on potential risk factors for SIDS were completed by parents of SIDS victims, and for at least two controls matched for gender, age and place of birth . All SIDS cases were verified by an autopsy . The study comprised 244 SIDS cases and 869 controls, analysed by conditional logistic regression . Significantly more cases than controls presenting symptoms of infectious diseases during the last week and/or last day were treated with antibiotics and had been seen by a physician . The finding is consistent with the hypothesis of an infectious mechanism in SIDS induced by local microorganism growth and toxin or cytokine production, and also adds further support to a possible association between infection and SIDS by loss of protective mechanisms, such as arousal . The risk of SIDS among infants with the combined presence of infectious symptoms and either of the other modifiable risk factors, prone sleeping, head covered or parental smoking, was far greater than the sum of each individual factor . These risk factors thus modify the dangerousness of infection in infancy. Structure Fold Des, 1999 Jul 15, 7(7), 865 - 75 Crystal structure of phenylalanine-regulated 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase from Escherichia coli; Shumilin IA et al.; BACKGROUND: In microorganisms and plants the first step in the common pathway leading to the biosynthesis of aromatic compounds is the stereospecific condensation of phosphoenolpyruvate (PEP) and D-erythrose-4-phosphate (E4P) giving rise to 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) . This reaction is catalyzed by DAHP synthase (DAHPS), a metal-activated enzyme, which in microorganisms is the target for negative-feedback regulation by pathway intermediates or by end products . In Escherichia coli there are three DAHPS isoforms, each specifically inhibited by one of the three aromatic amino acids . RESULTS: The crystal structure of the phenylalanine-regulated form of DAHPS complexed with PEP and Pb2+ (DAHPS(Phe)-PEP-Pb) was determined by multiple wavelength anomalous dispersion phasing utilizing the anomalous scattering of Pb2+ . The tetramer consists of two tight dimers . The monomers of the tight dimer are coupled by extensive interactions including a pair of three-stranded, intersubunit beta sheets . The monomer (350 residues) is a (beta/alpha)8 barrel with several additional beta strands and alpha helices . The PEP and Pb2+ are at the C-ends of the beta strands of the barrel, as is SO4(2-), inferred to occupy the position of the phosphate of E4P . Mutations that reduce feedback inhibition cluster about a cavity near the twofold axis of the tight dimer and are centered approximately 15 A from the active site, indicating the location of a separate regulatory site . CONCLUSIONS: The crystal structure of DAHPS(Phe)-PEP-Pb reveals the active site of this key enzyme of aromatic biosynthesis and indicates the probable site of inhibitor binding . This is the first reported structure of a DAHPS; the structure of its two paralogs and of a variety of orthologs should now be readily determined by molecular replacement. New Microbiol, 1999 Jul, 22(3), 181 - 6 Seroprevalence of anti-human parvovirus B19 antibodies in patients attending a centre for sexually transmitted diseases; Salvatori L et al.; The aim of this study was to establish the serological prevalence of anti-human Parvovirus B19 (HP-B19) antibodies in a group of 321 patients attending a Centre for Sexually Transmitted Diseases (STDs) and epidemiologically examine whether this virus may also be sexually transmitted . For this purpose, the serum prevalence of anti-HP-B19 evaluated in STD patients (39%) was compared with that of 164 healthy blood donors (10%, p < 0.001), using commercially available ELISA methods detecting the anti-VP1 reactivity of the sera . The same STD patients were also analyzed for serum reactivities against 4 STD-causing microorganisms, namely T . pallidum (TPHA), HBV (HBcAb), HCV (HCV-Ab) and HIV (HIV-Ab), to observe possible associations with the serum anti-HP-B19 reactivity . These tests were also carried out with commercially available kits . The results suggest that the serum anti-HP-B19 antibody prevalence in patients with STDs is increased, also independently of their intravenous drug addition and varies with the reactivity pattern determined . In addition, as expected for a STD, the anti-HP-B19 prevalence is increased in homobisexual patients compared with heterosexuals. Bacteriol Virusol Parazitol Epidemiol, 1998 Oct-Dec, 43(4), 237 - 9 {The development of an original reagent for the rapid determination of the deoxyribonuclease (DNAse) activity of microorganisms}; Morari N; The DN-ase test is the most important tests used for identification of pathogenic microorganisms . But often this test is not accessible for microbiology laboratories because of its high prices . The author elaborated a new original preparation for the rapid determination of the DN-ase activity, which permits a 350-400 fold reduction of the cost/test ratio . Moreover, the technique procedure is essentially simplified; there is