J Exp Biol, 1997 Dec, 200 ( Pt 23), 2977 - 86 A novel UDP-sugar, UDP-3-ketoglucosamine or UDP-4-ketoglucosamine, from bovine heart muscle reduces metmyoglobin with NAD(P)H; Kariya M et al.; 3-Ketoglucose and similar ketosugars have been identified in microorganisms only and little is known about their functions . UDP-sugars are widely found as an intermediate in sugar metabolism in living organisms . Yet what role UDP-sugars play, or whether they play a direct role in metabolism, is still unknown . UDP-sugars were isolated and purified from bovine heart muscle, and a UDP-sugar fraction capable of NAD(P)H-dependent catalytic reduction of metmyoglobin was detected . Subsequent identification revealed that the active UDP-sugar was UDP-3- or UDP-4-ketoglucosamine . These compounds were purified from bovine cardiac muscle by ultrafiltration, anion-exchange column chromatography and reverse-phase chromatography . They were further characterized by determination of their chemical reducing activity, by comparison with synthetic UDP-3- or UDP-4-ketoglucosamine standards using high-performance liquid chromatography, by estimation of molecular mass using fast atom bombardment mass spectrometry, and by Fourier transform infrared microspectroscopy and electron probe microanalysis . The results suggest that UDP-3- or UDP-4-ketoglucosamine reduces metmyoglobin in bovine cardiac muscle . It is important that the reducing activity displayed by this ketosugar is not the effect of UDP-3- or UDP-4-ketoglucosamine alone but depends on NAD(P)H . In other words, this action of UDP-3- or UDP-4-ketoglucosamine is catalytic. Adv Biochem Eng Biotechnol, 1998, 60, 153 - 94 Process models for production of beta-lactam antibiotics; Bellgardt KH; Great progress has been made in the modelling of biotechnical processes using filamentous microorganisms . This paper deals with cultivations of Penicillium chrysogenum for the production of Penicillin and of Acremonium chrysogenum for the production of Cephalosporin C . The properties of the processes and the existing models are reviewed . Models are presented for both processes that consider aspects which are important for industrial cultivation . The process model for Penicillin production is based on a detailed morphological description of growth of hyphal filaments and pellets . The model allows for simulation of the production process including the preculture and considering the inhomogenous pellet population . It opens new possibilities for understanding the complex kinetics of the process and improvement of its control . The structured segregated model for Cephalosporin C production considers soy oil as second carbon source besides sugar . The application of the model for dynamic optimization of feeding strategies by Iterative Dynamic Programming is demonstrated . As an alternative approach, modelling of the Cephalosporin production by an artificial neural network is discussed. Mycopathologia, 1997, 138(3), 127 - 35 The influence of surface carbohydrates on the interaction of Fonsecaea pedrosoi with Chinese hamster ovary glycosylation mutant cells; Limongi CL et al.; In order to better understand the role played by surface glycoconjugates during cell adhesion and endocytosis by the dematiaceous fungi Fonsecaea pedrosoi, we analyzed the interaction between this microorganism and five mutants of Chinese Hamster Ovary (CHO) cells, which differ from each other in the exposition of carbohydrate residues on the cell surface . Five clones (Gat-2 parental, and the clones: Lec1, Lec2, Lec8 and ldlLec1) were tested and the adhesion and endocytic indexes were determined after 2 hours of interaction . The Lec1 and ldlLec1 clones, which present exposed mannose residues, showed the greater adhesion index (AI) . On the other hand, the Lec8 clone, which exposes N-acetylglucosamine on the cell surface, presented the greater endocytic index . The role played by surface-exposed carbohydrate residues was further analyzed by addition of mannose or N-acetylglucosamine to the interaction medium and by previous incubation of the cells in the presence of the lectins Concanavalin A (ConA) and wheat germ agglutinin (WGA) . The results obtained suggest that mannose residues are involved in the first step of adhesion of F . pedrosoi to the cell surface, while N-acetylglucosamine residues are involved on its ingestion process. Oral Microbiol Immunol, 1997 Aug, 12(4), 219 - 26 Arbitrarily primed polymerase chain reaction fingerprinting and clonal analysis of oral Fusobacterium nucleatum isolates; George KS et al.; F . nucleatum is the most commonly isolated microorganism from subgingival plaque, but the role of this microorganism in periodontal diseases remains undefined . Arbitrarily primed polymerase chain reaction (AP-PCR) was evaluated as a method for fingerprinting F . nucleatum isolates and for use in clonal analysis . Pulsed field gel electrophoresis was used to further differentiate F . nucleatum isolates, with identical AP-PCR patterns . Extremely heterogeneous AP-PCR fingerprints were observed among the 98 F . nucleatum isolates, with 36 different genotypes observed with primer C1 and 30 different genotype detected with primer C2 . Combining the results of the AP-PCR genotype analysis from C1 and C2 primer amplifications revealed that up to 7 different genotypes could be distinguished from isolates from the same oral cavity and that up to 4 different genotypes were observed within a single site . An intense amplicon at approximately 450 bp generated in AP-PCR amplification with primer C2 was associated with F . nucleatum subsp . nucleatum (ATCC 25586) and with 15 F . nucleatum isolates from diseased sites and 2 isolates from healthy sites . Pulsed field gel electrophoresis confirmed the AP-PCR genotypes and demonstrated increased discriminatory power over AP-PCR . The results indicated that AP-PCR and pulsed field gel electrophoresis provide a simple and sensitive means for differentiating oral F . nucleatum isolates and further demonstrate the heterogeneity of this species . These techniques may serve as useful tools in the clonal and epidemiological analysis of F . nucleatum isolates, which may help define the role of these microorganisms in periodontal diseases. Aliment Pharmacol Ther, 1997 Dec, 11 Suppl 3, 3 - 8; discussion 8-9 Review article: Intestinal epithelia and barrier functions; Kraehenbuhl JP et al.; The mucosal epithelia of the digestive tract acts as a selective barrier, permeable to ions, small molecules and macromolecules . These epithelial cells aid the digestion of food and absorption of nutrients . They contribute to the protection against pathogens and undergo continuous cell renewal which facilitates the elimination of damaged cells . Both innate and adaptive defence mechanisms protect the gastrointestinal-mucosal surfaces against pathogens . Interaction of microorganisms with epithelial cells triggers a host response by activating specific transcription factors which control the expression of chemokines and cytokines . This host response is characterized by the recruitment of macrophages and neutrophils at the site of infection . Disruption of epithelial signalling pathways that recruit migratory immune cells results in a chronic inflammatory response . The adaptive defence mechanism relies on the collaboration of epithelial cells (resident sampling system) with antigen-presenting and lymphoid cells (migratory sampling system); in order to obtain samples of foreign antigen, these samples must be transported across the barriers without affecting the integrity of the barrier . These sampling systems are regulated by both environmental and host factors . Fates of the antigen may differ depending on the way in which they cross the epithelial barrier, i.e . via interaction with motile dendritic cells or epithelial M cells in the follicle-associated epithelium. Exp Gerontol, 1998 Jan-Mar, 33(1-2), 191 - 207 A novel technique for transforming the theft of mortal human cells into praiseworthy federal policy; Hayflick L; A revolution has occurred in the attitude of biologists toward their intellectual property rights . What today is patentable and highly profitable was, 20 years ago, unpatentable and given away for nothing . The history of this revolution began in the early 1960s when we made the first effort to have self-duplicating cell strains patented . The application was denied because patent law at that time did not include living matter . Because of the demand for our normal human diploid cell strain, WI-38, by NIH grantees, NIH support was provided to distribute WI-38 gratis to hundreds of recipients . These included vaccine and cell manufacturers who profited enormously from the direct sale of WI-38 or its use as a substrate for many human virus vaccines . When federal support for the distribution of WI-38 ended, but demand did not, I continued to distribute it for costs similar to those made by the American Type Culture Collection . When I took the first initiative and asked NIH to have the then unique question of title to a self-duplicating system resolved, they sent an accountant who accused me of theft of government property . I replied with a lawsuit that, after six years of litigation, we won with an out-of-court settlement . During these six years the United States Supreme Court ruled that living matter could be patented . Also, the biotechnology industry was launched by biologists who, like me, started companies using cells or microorganisms developed with federal support . This use of intellectual property rights by the nascent biotechnology industry was ultimately embraced by the entire biological community and by a directive from the President of the United States . This revolution has now evolved to the point where government biologists themselves may profit from research in federal laboratories, and the NIH itself aggressively seeks private commercial alliances . Universities have also pursued similar alliances to the extent that today the distinction between a research university and a commercial organization is only in the eyes of the Internal Revenue Service. Oral Microbiol Immunol, 1997 Aug, 12(4), 236 - 9 Transcriptional analysis of the groESL operon from Porphyromonas gingivalis; Hotokezaka H et al.; Transcriptional analysis of the groESL operon from Porphyromonas gingivalis, one of the obligative anaerobic oral microorganisms implicated in adult periodontitis, was performed . P . gingivalis 381 cultured at 37 degrees C was shifted to 42 degrees C, 45 degrees C or 48 degrees C for 10 mins . Northern hybridization analysis revealed that a band with 2.1-kb (kilo base pair) was observed, and the transcripts increased greatly by heat shock . Primer extension and S1 mapping detected four different 5'-ending sites of the mRNAs at the upstream region of the groES . Three sites out of the four were heat-inducible . There were inverted repeats and a Escherichia coli sigma 32-recognizing consensus sequence in the promoter region of the groESL, which may be relevant to the regulation of transcription of groESL operon in P . gingivalis . Both a heat shock promoter and inverted repeats may be relevant to the transcriptional regulation of the groESL operon in P . gingivalis. Mutat Res, 1997 Dec 12, 395(2-3), 215 - 21 Reinvestigation of in vivo genotoxicity studies in man . I . No induction of DNA strand breaks in peripheral lymphocytes after metronidazole therapy; Fahrig R et al.; Although a rodent carcinogen, metronidazole is widely used in humans for the treatment of infections with anaerobic organisms . Metronidazole is mutagenic for microorganisms, but has a mainly negative data base for mammals and humans . Therefore, metronidazole is generally considered as a non-genotoxic carcinogen . Only the results of two human in vivo studies would allow the classification of metronidazole as genotoxic carcinogen: (1) the induction of DNA strand breaks; and (2) the induction of chromosome aberrations in peripheral lymphocytes after metronidazole therapy . Because the classification of metronidazole as genotoxic carcinogen would imply enormous consequences with respect to its application, both studies were reinvestigated very thoroughly . The present report describes the reinvestigation of the induction of DNA strand breaks after metronidazole therapy . Each two probes of lymphocytes of metronidazole-treated patients (3 x 500 to 3 x 750 mg/day for 5-8 days) were examined separately for the appearance of DNA strand breaks before and after treatment . In total, 400 nuclei were examined per patient . Immediately before the first, and 30 min to 2 h after the last application, 2 x 10 ml blood per patient was sampled, transported to the laboratory at 15-20 degrees C to make DNA repair more difficult, and examined within the next 4-7 h for DNA strand breaks . At the same time, the individual metronidazole blood plasma levels were measured . In contrast to the published reports, no induction of DNA strand breaks after metronidazole therapy could be observed in the present study . As the applied doses (15,750 mg vs . 4800 mg) and the plasma level (up to 25 micrograms/ml vs . not measured) of metronidazole were much higher than in the published study, the relevance of the clearly negative result is obvious . As induction of DNA strand breaks is a frequent prerequisite for genotoxicity, metronidazole should be considered as a non-genotoxic carcinogen, and not as a genotoxic carcinogen. J Trauma, 1998 Jan, 44(1), 171 - 4 The presence of hemorrhagic shock increases the rate of bacterial translocation in blunt abdominal trauma; Kale IT et al.; BACKGROUND: Sepsis and multisystem organ failure are common after hemorrhagic shock . The aims of this study were to determine whether hemorrhagic shock would promote the translocation of bacteria and if it correlates with clinical outcome in patients with blunt abdominal trauma . METHODS: Twenty-six patients requiring laparotomy for blunt abdominal trauma (group I) and 30 patients operated electively (group II) were studied . Injury Severity Score, Trauma Score, and Acute Physiology and Health Evaluation (APACHE) II score were recorded before the operation . Peritoneal swab, mesenteric lymph node, portal venous blood, liver wedge biopsy, and spleen biopsy (in splenectomized patients) were sampled for culture after surgical hemostasis . Additionally, peripheral blood samples were taken preoperatively and postoperatively in group I patients for culture . The same samples were taken in group II patients except for the spleen biopsy . Moreover, patients in group I were further subdivided into subgroups A and B, indicating the presence or absence, respectively, of hemorrhagic shock (defined as systolic blood pressure < 90 mm Hg with identifiable blood loss) . Postoperatively, patients were checked for infectious and septic complications . RESULTS: Mean Injury Severity Score, Trauma Score, and APACHE II score were 32.0, 12.1, and 10.9 in group I and 2.1 (APACHE II,p < 0.01) in group II, respectively . Two patients in group IA, eight patients in group IB, and one patient in group II demonstrated bacterial translocation (BT) (p < 0.01) . Five patients with blunt abdominal trauma had major infectious complications, but only one had BT, and the same microorganism grew in the intra-abdominal abscess . There were two infectious complications in the control group . One of these patients had BT, and the same microorganism grew in the wound infection . CONCLUSION: We conclude that BT occurs after blunt abdominal trauma in humans and correlates with the presence of hemorrhagic shock, but the clinical significance of BT in trauma patients remains unclear. J Philipp Dent Assoc, 1996 Sep-Nov, 48(2), 31 - 8 The effect of 1% povidone iodine as a pre-procedural mouthrinse in 20 patients with varying degrees of oral hygiene; Domingo MA et al.; This study is conducted to determine the different types of microorganisms found in the saliva of individuals with varying degrees of oral hygiene, also to determine the effectiveness of 1% Povidone Iodine (Betadine) 1% gargle oral antiseptic as a pre-procedural mouthrinse in individuals with varying degrees of oral hygiene, and lastly to be able to determine the duration of the effectiveness of the solution . The cheek mucosa of the patients are procured of saliva specimens without prior rinsing of 1% Povidone Iodine and the saliva sample obtained was placed on a test tube with thioglycollate culture media and was labelled . The patients are asked to rinse or to gargle immediately with 1% Povidone Iodine at full strength (20ml.) for about 30 seconds . After two hours, the cheek mucosa was then again swabbed and placed on the second test tube . After four hours, the same procedure was done and the saliva specimen obtained was labelled and placed on the third test tube . The result of this study revealed that 1% Povidone Iodine used as a pre-procedural mouthrinse has a bactericidal effect in the microorganism concentration resulting to the reduction of surviving microorganisms up to four hours which is the limitation of the study. Pol Merkuriusz Lek, 1997 Oct, 3(16), 208 - 9 {Chlamydia pneumoniae as an etiologic factor in disease of the respiratory tract}; Nalepa P; The described recently species of Chlamydia pneumoniae is the subject of a large number of scientific reports . The majority of infections caused by this microorganism are asymptomatic . However, a lot of cases of symptomatic infections have been described with both mild and severe clinical course . Symptoms related to infection may affect both upper and lower parts of the respiratory tract and may be accompanied by disorders in other systems and organs . Reports on the influence of chronic inflammation by Chlamydia pneumoniae on the development of arteriosclerosis are still controversial . The effective drugs in the treatment of Chlamydia pneumoniae infections are macrolides and tetracyclines. Am J Clin Nutr, 1998 Feb, 67(2), 332 - 7 Safflower oil consumption does not increase plasma conjugated linoleic acid concentrations in humans; Herbel BK et al.; Conjugated linoleic acid (CLA) is a mixture of positional and geometric isomers of linoleic acid (LA) with conjugated double bonds . CLA has anticarcinogenic properties and has been identified in human tissues, dairy products, meats, and certain vegetable oils . A variety of animal products are good sources of CLA, but plant oils contain much less . However, plant oils are a rich source of LA, which may be isomerized to CLA by intestinal microorganisms in humans . To investigate the effect of triacylglycerol-esterified LA consumption on plasma concentrations of esterified CLA in total lipids, a dietary intervention (6 wk) was conducted with six men and six women . During the intervention period a salad dressing containing 21 g safflower oil providing 16 g LA/d was added to the subjects' daily diets . Three-day diet records and fasting blood were obtained initially and during dietary and postdietary intervention periods . Although LA intake increased significantly during the dietary intervention, plasma CLA concentrations were not affected . Plasma total cholesterol and LDL-cholesterol concentrations were significantly lower after addition of safflower oil to the diet . In summary, consumption of triacylglycerol-esterified LA in safflower oil did not increase plasma concentrations of esterified CLA in total lipids. Diagn Microbiol Infect Dis, 1997 Dec, 29(4), 219 - 25 Comparison of BACTEC plus Aerobic/F, Anaerobic/F, Peds Plus/F, and Lytic/F media with and without fastidious organism supplement to conventional methods for culture of sterile body fluids; Fuller DD et al.; We compared the BACTEC 9240 continuous-read instrument using Peds Plus/F, Lytic/F, Aerobic/F, and Anaerobic/F media (Becton Dickinson Diagnostic Instrument Systems, Sparks, MD) with and without fastidious organism supplement to conventional centrifugation preparation and plating for the recovery and speed of detection of microorganisms . A total of 908 sterile body fluid specimens were collected and processed, yielding 116 (13%) positive cultures . Of the 80 isolates considered clinically significant, 48 (60%) were recovered by both the BACTEC system and conventional culture, whereas 32 (40%) were recovered by BACTEC only . No clinically significant isolates were recovered only by conventional culture methods . The time to detection for isolates recovered from both sets was faster for BACTEC . It was found that BACTEC, with or without the addition of fastidious organisms supplement, exhibited improved sensitivity for the recovery of microorganisms. J Am Podiatr Med Assoc, 1998 Jan, 88(1), 17 - 20 Transmission of microbial pathogens by cedar shoe trees; Woeste S; Cedar shoe trees are used to keep shoes from shrinking and losing their shape . When the same cedar shoe trees are used by different shoe wearers, there is a chance of transmission of disease-causing microorganisms between people . This study was conducted to determine whether or not transmission of disease-causing microorganisms via cedar shoe trees occurs, what kinds of microorganisms can be transmitted, and how many organisms can be transmitted . In this study, both bacteria and bacterial spores were transmitted, while fungi were not; however, only several hundred to several thousand pathogenic bacteria were transmitted between any two shoes. Zh Mikrobiol Epidemiol Immunobiol, 1997 Sep-Oct, (5), 22 - 5 {The epidemiological aspects of a pseudotuberculosis polyclonal infection}; Shubin FN; The plasmid profile of 489 Y . pseudotuberculosis strains, serovar 1b, isolated from patients during 23 outbreaks and in sporadic cases of the disease were studied . As revealed in this study, in foci with the heterogeneous population of Y . pseudotuberculosis, serovar 1b, outbreaks and sporadic cases of the disease were caused by the polyclonal population of the microorganism . The study showed that three types of outbreaks could be differentiated, depending on the clonal characteristics of the microorganism causing these outbreaks and the presence of the dominating clone of the causative agent . Y . pseudotuberculosis infection caused by the combination of several clones of the microorganism was designated as polyclonal infection. Rocz Panstw Zakl Hig, 1997, 48(3), 269 - 74 {The action of ultraviolet radiation on microorganisms found in air}; Krzywicka H et al.; The effect of a device with UV lamp on the microorganisms in the air was investigated . The air was continuously pumped through the device for 24 h . After that time the number of microorganisms was reduced by 0.3 log. Helicobacter, 1997 Jun, 2(2), 100 - 5 Catalase and superoxide dismutase secreted from Helicobacter pylori; Mori M et al.; BACKGROUND: Previously, we have reported that toxic oxidants produced by activated neutrophils play a pivotal role in the development of Helicobacter pylori-induced gastric mucosal damage . Microorganisms, however, are characterized by their ability to produce a variety of antioxidant enzymes . This study is designed to measure the oxygen radical scavenging enzymes, such as catalase and superoxide dismutase (SOD), and urease in the supernatant of H . pylori (NCTC11637) suspension . MATERIALS AND METHODS: H . pylori was inoculated on a sheep blood agar plate and harvested . Bacterial suspensions (10(9) cfu/ml phosphate buffer) were washed twice and incubated at 37 degrees C for 1, 2, 12, and 24 hours or were sonicated in ice . Their supernatants were obtained by centrifugation and filtration . SOD activity was measured spectrophotometrically by the cytochrome c method . Catalase activity was assayed by the fall in absorbance at 240 nm as H2O2 is degraded . Urease activity was determined by measuring the release of ammonia using Berthelot reaction . RESULTS: Activities of both SOD and catalase were detected in the supernatants of 1-hour microaerophilic incubation . Their activities were almost constant in 4-, 12-, and 24-hour microaerobic incubation or sonication . Urease activity was increased dramatically in proportion to the period of microaerobic incubation . CONCLUSION: Although H . pylori possesses antioxidant as a constitutive compartment, the magnitude of its secretion was below the detectable level . It is not likely that SOD and catalase play a significant role for scavenging the oxidants from injured gastric mucosa, such as infiltrated leukocytes. J Clin Microbiol, 1998 Jan, 36(1), 105 - 9 Earlier positivity of central-venous- versus peripheral-blood cultures is highly predictive of catheter-related sepsis; Blot F et al.; To diagnose catheter-related sepsis without removing the catheter, we evaluated the differential positivity times of cultures of blood drawn simultaneously from central venous catheter and peripheral sites . In a 450-bed cancer reference center, simultaneous central- and peripheral-blood cultures were prospectively performed for patients with suspicion of catheter-related sepsis over an 18-month period . Data for 64 patients for whom the same microorganisms were found when central- and peripheral-blood samples were cultured were retrospectively reviewed by two independent physicians blinded to the differential positivity time values in order to establish or refute the diagnosis of catheter-related sepsis . The diagnosis was established in 28 cases, refuted in 14, and indeterminate in the remaining 22 . The differential positivity time was significantly greater for patients with catheter-related sepsis (P < 10(-4)) . A cutoff limit of +120 min had 100% specificity and 96.4% sensitivity for the diagnosis of catheter-related sepsis . These results strongly suggest that measurement of the differential positivity time might be a reliable tool facilitating the diagnosis of catheter-related sepsis in patients with an indwelling catheter. J Clin Microbiol, 1998 Jan, 36(1), 77 - 80 Clinical evaluation of a new PCR assay for detection of Coxiella burnetii in human serum samples; Zhang GQ et al.; A nested PCR method was developed for the detection of Coxiella burnetii in human serum samples . Two pairs of oligonucleotide primers were designed to amplify a 438-bp fragment of the com1 gene encoding a 27-kDa outer membrane protein of C . burnetii . The primers amplified the predicted fragments of 21 various strains of C . burnetii but did not react with DNA samples from other microorganisms . The 438-bp amplification products could be digested with restriction enzymes SspI and SalI . The utility of the nested PCR was evaluated by testing human serum samples . The com1 gene fragment was amplified from 135 (87%) of 155 indirect immunofluorescence test (IF)-positive serum samples and from 11 (11%) of 100 IF-negative serum samples . The nested PCR with primers targeted to the com1 gene appeared to be a sensitive, specific, and useful method for the detection of C . burnetii in serum samples. J Biol Chem, 1997 Dec 12, 272(50), 31301 - 7 A novel white laccase from Pleurotus ostreatus; Palmieri G et al.; Two laccase isoenzymes (POXA1 and POXA2) produced by Pleurotus ostreatus were purified and fully characterized . POXA1 and POXA2 are monomeric glycoproteins with 3 and 9% carbohydrate content, molecular masses of about 61 and 67 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis, of about 54 and 59 kDa by gel filtration in native conditions, and of 61 kDa by matrix-assisted laser desorption ionization mass spectrometry (only for POXA1) and pI values of 6.7 and 4.0, respectively . The N terminus and three tryptic peptides of POXA1 have been sequenced, revealing clear homology with laccases from other microorganisms, whereas POXA2 showed a blocked N terminus . The stability of POXA2 as a function of temperature was particularly low, whereas POXA1 showed remarkable high stability with respect to both pH and temperature . Both enzymes oxidize syringaldazine and ABTS (2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) together with a variety of different substituted phenols and aromatic amines with the concomitant reduction of oxygen, but POXA1 is unable to oxidize guaiacol . Both enzymes were strongly inhibited by sodium azide and thioglycolic acid but not by EDTA . UV/visible absorption spectra, atomic adsorption, and polarographic data indicated the presence of 4 copper atoms/mol of POXA2 but only one copper, two zinc, and one iron atoms were found/mol of POXA1 . The neutral pI and the anomalous metal content of POXA1 laccase render this enzyme unique in its structural characteristics . The lack of typical absorbance at 600 nm allows its classification as a "white" laccase. J Clin Invest, 1997 Dec 1, 100(11), 2737 - 43 Effects of subcutaneous interleukin-2 therapy on CD4 subsets and in vitro cytokine production in HIV+ subjects; De Paoli P et al.; HIV infection is characterized by the reduction of the CD4+, CD45RA+, CD26+, and CD28+ lymphocyte subsets and of the in vitro production of IL-2, IL-4, and interferon-gamma; on the contrary, chemokine production is usually increased . These abnormalities are only partially restored by antiretroviral chemotherapy . Therapy with interleukin-2 has been proposed to restore the functions of the immune system, but the mechanisms by which IL-2 exerts its activities are unknown . The aim of this study was to define the effects of rIL-2 administration on CD4+, CD45RA+, CD45R0+, and CD26+ lymphocytes and on the in vitro production of IL-2, IL-4, IL-10, IFN-gamma, RANTES, and sCD30 in HIV+ patients . 10 HIV+ patients with CD4 cell counts between 200 and 500 cells/mm3 were treated with six cycles of subcutaneous recombinant IL-2 administration, in combination with zidovudine and didanosine . This therapeutic regimen resulted in a remarkable increase in the number of CD4+ cells and in the prolonged reduction of the levels of viremia . CD45R01 cells were expanded during the first cycle of therapy, while CD45RA+/CD26+ cells predominated after the third cycle . At this time, the in vitro production of IL-2, IL-4, IFN-gamma, and sCD30 were significantly upregulated . These results demonstrate that rIL-2 in HIV+ patients induces the reconstitution of the CD4/CD45RA lymphocytes subtype . This expanded cell population recovered the ability to produce in vitro IL-2, IL-4, and IFN-gamma . These effects may be beneficial to HIV+ patients by improving their immune response to microorganisms or vaccines. Zh Mikrobiol Epidemiol Immunobiol, 1995 Jul-Aug, (4), 40 - 2 {Infectious risk factors in the development of malignant neoplasms}; Shevliagin VIa et al.; The influence of microorganisms on the course of tumor processes in laboratory animals was studied . The study revealed that the inoculation of Bordetella pertussis decreased survival time both in mice with implanted mastocytoma cells P-815 and in Syrian golden hamsters with implanted transformed hamster fibroblastomas . The smallpox vaccine virus was found to stimulate the development of virus-induced Rauscher leukosis in mice . The mechanisms of the co-carcinogenic action of microorganisms is discussed. Int J Biochem Cell Biol, 1997 Jul, 29(7), 939 - 43 Fibronectin; Romberger DJ; Fibronectin is a glycoprotein consisting of repeating units of amino acids, which form domains that enable the molecule to interact with a variety of cells through both integrin and non-integrin receptors . It is encoded by a single gene, but alternative splicing of pre-mRNA allows formation of multiple isoforms that have critical roles in cell adhesion, migration and proliferation . The essential nature of fibronectin in development has clearly been demonstrated in a "knock-out" mouse model in which early lethality occurs . Fibronectin influences diverse processes including inflammation, wound repair, malignant metastasis, microorganism attachment and thrombosis . Researchers are currently developing tools, including synthetic peptides based on specific fibronectin regions . These molecules have been shown to alter processes such as lymphocyte binding in synovial tissue of rheumatoid arthritis, coronary arteriopathy in animal models of cardiac transplantation, and platelet aggregation in patients, and are thus providing important new therapeutic possibilities. Clin Diagn Lab Immunol, 1998 Jan, 5(1), 11 - 7 A novel approach for detecting an immunodominant antigen of Porphyromonas gingivalis in diagnosis of adult periodontitis; Kawai T et al.; In the course of long-term infection with Porphyromonas gingivalis in adult periodontitis, a specific antibody response to this organism is generated . We describe a potential novel approach for identifying an immunodominant antigen in human periodontitis patients . First, various monoclonal antibodies (MAbs) were established from mice immunized with crude antigen preparations of P . gingivalis FDC 381 . The antigen specificities of these MAbs were compared with those of serum antibodies of 10 periodontitis patients in a competitive enzyme-linked immunosorbent assay . The binding of one MAb (termed PF18) was readily inhibited by sera from all patients but not by sera from healthy volunteers . The antigen recognized by PF18 existed on the cell surface, presumably in the capsule layer, shown by immunoelectron microscopic analysis . Purification of the antigenic substance, termed PF18-Ag, was performed by immunoaffinity chromatography with the MAb . Characterization of PF18-Ag suggested that the epitope was composed of carbohydrates but not peptides and that the substance was different from lipopolysaccharide . Measurement of levels of serum antibody to PF18-Ag better discriminated periodontitis patients from healthy individuals than measurement of antibodies to crude antigen preparations of P . gingivalis . Immunoglobulin G2 was the predominant isotype among the antibodies to PF18-Ag in the patients' sera . These results suggest that PF18-Ag, which is possibly a novel substance, is an important antigenic substance and is potentially useful for the clinical diagnosis of adult periodontitis . The approach that was used would also be relevant to detecting immunodominant antigens of other infectious microorganisms. FEMS Microbiol Lett, 1998 Jan 1, 158(1), 51 - 5 Distribution of cyclic imide-transforming activity in microorganisms; Soong CL et al.; Cyclic imide-transforming activity was found to be widely distributed in bacteria, yeast and molds . This activity was not correlated with cyclic ureide-transforming activity in bacteria, but there was some correlation in yeast and molds . These two activities are probably catalyzed by different enzymes in bacteria . Besides the well-known cyclic ureide transformation, cyclic imide transformation by microorganisms was common. J Calif Dent Assoc, 1997 Mar, 25(3), 215 - 8 Microleakage of endodontic access cavities restored with composites; Barkhordar RA et al.; A proper seal of restorative composites in endodontic access cavities is mandatory to prevent ingress of microorganisms and debris into the tooth-restoration interface . Chemically cured composites tend to have less polymerization shrinkage than photopolymerization composites . This study compared in vitro sealing performance of a chemically cured and a photopolymerization composite for access cavity restorations in endodontically treated teeth . In addition, the effects of four dentin pretreatments on the seal were evaluated . One hundred twenty-eight roots of extracted human incisors were cleansed, shaped and obturated with gutta percha and Grossman's sealer . The endodontic access cavities were later treated by Scotchbond Multipurpose, All-Bond 2, Vitrebond and Ketac-Bond . Half the access cavities within each group were filled by Coltene Microhybrid H.P., a chemically cured microhybrid composite . The other half were filled with Aelitfil, a photopolymerized composite . The filled teeth were thermocycled, submerged in silver nitrate solution, sectioned longitudinally, and their dye penetration measured and statistically analyzed by two-way ANOVA at 95 percent confidence level . The results show that the chemically cured composite allowed less dye penetration than the photopolymerized composite . As for the dentin pretreatments, the Scotchbond Multipurpose and All-Bond 2 exhibited less microleakage than Vitrebond and Ketac-Bond for both chemical and photopolymerized composite . Ketac-Bond allowed the highest amount of dye penetration, followed by Vitrebond, Scotchbond Multipurpose, then All-Bond, which had the least depth of microleakage . Chemically cured composites bonded with All-Bond 2 and Scotchbond Multipurpose may exhibit less microleakage than photopolymerized composites in endodontic access cavity restoration. Histochem Cell Biol, 1997 Dec, 108(6), 455 - 70 The role of M cells in the protection of mucosal membranes; Gebert A; The mucosa-associated lymphoid tissues continuously take up antigenic matter from the lumen to generate immune responses or to maintain immune tolerance . This antigen sampling is performed by highly specialised epithelial cells, the membranous (M) cells of the dome epithelia . M cells possess a unique ultrastructure and lie in close contact to lymphoid cells . They endocytose soluble and solid substances, including entire microorganisms, at their apical membrane, transport these in vesicles to their basolateral membrane and exocytose them to the intercellular space . This review summarises the structural and functional peculiarities of M cells in different species and at the different sites of lymphoid tissue along the digestive and respiratory tracts . Specialisations of M cells for antigen uptake and transport comprise the composition of their apical membrane and its glycocalyx, a modified cytoskeleton as compared to enterocytes and a pocket-like invagination of the basolateral membrane populated by lymphocytes and macrophages . Besides ultrastructural characteristics, histochemical markers are listed that are currently available for detecting M cells by light microscopy . The origin, differentiation and distribution of M cells and other epithelial cell types of the dome epithelium are outlined . As M cells are used as entry sites by various pathogens and, in the future, might be employed for the oral application of vaccines and drugs, the clinical relevance of M cells in health and disease is discussed. J Am Acad Dermatol, 1998 Jan, 38(1), 56 - 60 A new application of BCG antibody for rapid screening of various tissue microorganisms; Kutzner H et al.; BACKGROUND: In routine dermatopathology there is growing demand for a simple, fast, cost-effective, and highly sensitive screening tool for the detection of microorganisms . OBJECTIVE: Our purpose was to determine whether immunostaining with polyclonal anti-Mycobacterium bovis (BCG), which is known for its interspecies cross-reactivity, is a suitable screening method for many common microorganisms in dermatopathologic specimens . METHODS: A total of 254 formalin-fixed, paraffin-embedded skin specimens of viral, protozoal, fungal, and bacterial infections were stained with appropriate histochemical stains and with anti-BCG . RESULTS: Anti-BCG labeled bacteria and fungi with high sensitivity and minimal background staining, but did not react with spirochetes, viruses, or protozoa (Leishmania) . The quality and sensitivity of anti-BCG staining were superior to conventional histochemical stains . CONCLUSION: Because of its cross-reactivity with many bacteria and fungi as well as its high sensitivity and minimal background staining, the anti-BCG immunostain is a promising screening tool for the detection of the most common bacterial and fungal microorganisms in paraffin-embedded skin specimens. Blood, 1998 Feb 1, 91(3), 1044 - 58 Azurophilic granules of human neutrophilic leukocytes are deficient in lysosome-associated membrane proteins but retain the mannose 6-phosphate recognition marker; Cieutat AM et al.; During granulocyte differentiation in the bone marrow (BM), neutrophilic leukocyte precursors synthesize large amounts of lysosomal enzymes . These enzymes are sequestered into azurophilic storage granules until used days later for digestion of phagocytized microorganisms after leukocyte emigration to inflamed tissues . This azurophil granule population has previously been defined as a primary lysosome, i.e., a membrane-bound organelle containing acid hydrolases that have not entered into a digestive event . In this study, azurophil granules were purified and shown to contain large amounts of mannose 6-phosphate-containing glycoproteins (Man 6-P GP) but little lysosome-associated membrane proteins (LAMP) . In addition, the fine structural localization of Man 6-P GP and LAMP was investigated at various stages of maturation in human BM and blood . Man 6-P GP were present within the azurophilic granules at all stages of maturation and in typical multivesicular bodies (MVB) as well as in multilaminar compartments (MLC), identified by their content of concentric arrays of internal membranes . LAMP was absent in all identified granule populations, but was consistently found in the membranes of vesicles, MVB, and MLC . The latter compartment has not been previously described in this cell type . In conclusion, the azurophilic granules, which contain an abundance of lysosomal enzymes and Man 6-P GP, lack the LAMP glycoproteins . By current criteria, they therefore cannot be classified as lysosomes, but rather may have the functional characteristics of a regulated secretory granule . Rather, the true lysosomes of the resting neutrophil are probably the MVB and MLC . Finally, the typical "dense bodies" or mature lysosomes described in other cells are not present in resting neutrophils. J Aerosol Med, 1997 Summer, 10(2), 105 - 16 The size distribution of droplets in the exhaled breath of healthy human subjects; Papineni RS et al.; Droplets carried in exhaled breath may carry microorganisms capable of transmitting disease over both short and long distances . The size distribution of such droplets will influence the type of organisms that may be carried as well as strategies for controlling airborne infection . The aim of this study was to characterize the size distribution of droplets exhaled by healthy individuals . Exhaled droplets from human subjects performing four respiratory actions (mouth breathing, nose breathing, coughing, talking) were measured by both an optical particle counter (OPC) and an analytical transmission electron microscope (AEM) . The OPC indicated a preponderance of particles less than 1 mu, although larger particles were also found . Measurements with the AEM confirmed the existence of larger sized droplets in the exhaled breath . In general, coughing produced the largest droplet concentrations and nose breathing the least, although considerable intersubject variability was observed. J Clin Invest, 1997 Dec 15, 100(12), 3014 - 8 Immunization against the agent of human granulocytic ehrlichiosis in a murine model; Sun W et al.; The agent of human granulocytic ehrlichiosis (HGE) is a newly recognized tick-borne pathogen that resides within polymorphonuclear leukocytes . C3H/HeN mice can become infected with the agent of HGE (designated aoHGE) by syringe inoculation or tick-borne infection and develop transient neutropenia . They thereby partially mimic human disease and provide a model in which to study immunity to this microorganism . Mice vaccinated with lysates of purified aoHGE, or administered aoHGE antisera, were partially protected from both syringe- and tick-transmitted challenge with aoHGE . These data suggest that antibodies are sufficient to provide substantial, but not complete, immunity against aoHGE. Annu Rev Entomol, 1998, 43, 449 - 69 Parasites and pathogens of mites; Poinar G Jr et al.; Mites are an ancient group of arachnids that have had some 400 million years to adapt to a variety of conditions on Earth . Microorganisms have had the same amount of time to form symbiotic relationships with mites, with results ranging from phoresy to parasitism . This review covers the still fragmentary information on the groups of parasites and pathogens that are associated with mites . The known mite-associated bacteria, rickettsiae, fungi, Protozoa, viruses, and nematodes represent the tip of the iceberg, and few details of their host-parasite relationships have been recorded . Mites offer an opportunity to investigate new pathogens and new types of associations . Pathogens can be a boon when they affect mites that are detrimental to crops, livestock, or ourselves, and the diseases they cause probably play an important role in controlling mites, at least under certain conditions . However, pathogens can also cause crop failure and economic loss when they occur in biological control agents of pests. J Med Virol, 1998 Jan, 54(1), 44 - 53 Heparin-binding domain of human fibronectin binds HIV-1 gp120/160 and reduces virus infectivity; Bozzini S et al.; In vitro experiments indicate that components of the host present in body fluids may prevent the attachment of human immunodeficiency virus type 1 (HIV-1) to target cells . Fibronectin (Fn), a dimeric 440-kDa extracellular matrix adhesion protein, is secreted by mesenchymal cells and assembled into insoluble matrices . Fn exerts important effects on cell growth and differentiation through a number of discrete functional domains . Several microorganisms are known to bind Fn . We show that, under physiological conditions, HIV-1 gp120 and gp160 are capable of binding plasma and cellular Fn as well as laminin and vitronectin . Experiments were set up to analyze in detail the binding of HIV gp120 and gp160 to Fn . The gp120 and gp160 specifically recognize the C-terminal heparin-binding domain of Fn (Fn-CTHBD) with a calculated KD of 2.8 x 10(-7) M for gp160 . Binding of gp160 to Fn-CTHBD is a saturable and specific process that is blocked by antibodies to Fn-CTHBD and by heparin and is inhibited to a minor extent by heparan sulfate and dextran sulfate . These observations suggest that gp120/160 specifically recognize the III15 repeat within Fn-CTHBD . Intact Fn and Fn-CTHBD strongly inhibit the interaction of gp120/160 with soluble CD4 and, under low serum conditions, are capable of neutralizing the infectivity of HIV-1 for CD4-positive T cells . Thus, Fn that is present in plasma and mucinous secretions may well affect HIV infectivity and virus distribution in vivo. Pediatr Dent, 1997 Nov-Dec, 19(8), 476 - 9 Comparison of odontogenic and nonodontogenic facial cellulitis in a pediatric hospital population; Unkel JH et al.; Facial cellulitis in the pediatric hospital population can be classified as odontogenic and nonodontogenic . Emergency departments welcome timely diagnosis from consultants as cellulitis is associated with significant morbidity in children . The purpose of this retrospective study is to assist pediatric dentists in recognizing differences between odontogenic and nonodontogenic facial cellulitis and to determine whether odontogenic infections make up a major portion of facial swellings seen upon admission to the hospital . The completed medical records of 100 patients admitted to Children's Hospital of Pittsburgh from 1980-1989 with an ICD-9 diagnosis of facial cellulitis were reviewed . The types of cellulitis were differentiated using admission data . The information reviewed included age, sex, temperature, white blood cell count, location of facial infection, and season of the year . Odontogenic cellulitis comprised approximately 50% of the total hospital facial infections of the records reviewed during the 10-year period . Upon admission, patients with odontogenic and nonodontogenic facial cellulitis have similarities (season of onset during the year, febrile temperature, and location of infection) and differences (mean admission temperature, age at time of affliction, white blood cell count, and most commonly occurring microorganisms. Tidsskr Nor Laegeforen, 1997 Nov 10, 117(27), 3957 - 60 {New diagnostic and therapeutic possibilities in neonatal sepsis}; Hogasen AK; Neonatal sepsis remains a major clinical problem in neonatology, with high morbidity and mortality rates . The host defence against infections is immature in the newborn infant, and this makes the child more susceptible to invasive infection . The neutrophil storage pool and various granulocyte functions are impaired . In addition, the levels of immunoglobulins and complement are low . The detection of raised levels of complement activation products and cytokines may be of diagnostic help at an early stage of neonatal infection . Rapid treatment with antibiotics is essential for a favourable outcome . Possible adjuvant treatment may be to reduce the relative immunodeficiency by giving immunoglobulins or colony-stimulating factors which increase the production of leukocytes . Further, the potent inflammatory reaction initiated by the microorganisms may be suppressed by various therapies . In spite of much research in this field, no such adjuvant treatment has so far been shown to improve the outcome of neonatal sepsis. J Bacteriol, 1998 Jan, 180(2), 231 - 5 Chemotaxis in Borrelia burgdorferi; Shi W et al.; Borrelia burgdorferi is a motile spirochete which has been identified as the causative microorganism in Lyme disease . The physiological functions which govern the motility of this organism have not been elucidated . In this study, we found that motility of B . burgdorferi required an environment similar to interstitial fluid (e.g., pH 7.6 and 0.15 M NaCl) . Several methods were used to detect and measure chemotaxis of B . burgdorferi . A number of chemical compounds and mixtures were surveyed for the ability to induce positive and negative chemotaxis of B . burgdorferi . Rabbit serum was found to be an attractant for B . burgdorferi, while ethanol and butanol were found to be repellents . Unlike some free-living spirochetes (e.g., Spirochaeta aurantia), B . burgdorferi did not exhibit any observable chemotaxis to common sugars or amino acids . A method was developed to produce spirochete cells with a self-entangled end . These cells enabled us to study the rotation of a single flagellar bundle in response to chemoattractants or repellents . The study shows that the frequency and duration for pausing of flagella are important for chemotaxis of B . burgdorferi. Am J Infect Control, 1997 Dec, 25(6), 471 - 6 Natural bioburden levels detected on rigid lumened medical devices before and after cleaning; Chan-Myers H et al.; Controversy exists concerning the degree of microbial contamination associated with the use of rigid lumened medical devices, the efficacy of standard cleaning techniques used to remove pathogenic microorganisms from lumen channels, and whether patients are placed at risk of cross infection because of microbial contamination . In this study the level and types of microorganisms found on rigid lumened medical devices before and after cleaning in a hospital environment were investigated . The bioburden level after clinical use was found to be relatively low, ranging from 10(1) to 10(4) colony forming units (CFU) per device . After the instruments were cleaned, none of the devices studied contained bioburden levels greater than 10(4) CFU and 83% had bioburden levels less than or equal to 10(2) CFU . The bioburden present before cleaning was comprised of organisms derived from the handling of the device, from the hospital environment, and from the patient . The bioburden present after cleaning was comprised of organisms typically derived from the handling of the device and from the hospital environment . The level of bioburden per device was also related to the anatomic site where the device was used, with lower numbers of organisms found on devices exposed to sterile body sites and the respiratory tract. Acta Obstet Gynecol Scand, 1997 Nov, 76(10), 964 - 8 Relationship of bacteriological characteristics to semen indices and its influence on fertilization and pregnancy rates after IVF; Bussen S et al.; BACKGROUND: To determine if routine semen culture is useful in asymptomatic couples joining an in-vitro fertilization (IVF/ET) program . METHODS: Bacterial cultures and semen analysis according to WHO recommendations were performed on semen samples obtained before oocyte recovery from 88 asymptomatic couples undergoing IVF during a 7-month period . RESULTS: In 46 cultures at least one kind of microorganisms could be isolated . Forty-two cultures either contained bacterias regarded as normal skin flora (n = 14) or showed no growth of microorganisms (n = 28) . No differences were found in sperm concentration, total sperm count and sperm morphology between the semen samples with positive bacteriology and those with negative culture results . Sperm motility was decreased before Percoll preparation if microorganisms were present . Positive culture results had no effect on either fertilization or pregnancy rates . CONCLUSIONS: These observations suggest that bacteriospermia is not associated with abnormal sperm function after Percoll preparation or adverse IVF outcome. Gene, 1997 Dec 19, 204(1-2), 115 - 20 Cloning and characterization of human CIRP (cold-inducible RNA-binding protein) cDNA and chromosomal assignment of the gene; Nishiyama H et al.; Cold stress induces in microorganisms the synthesis of several proteins that are involved in various cellular processes such as transcription, translation and recombination . Recently, the cold-inducible RNA-binding protein (Cirp) was found to be induced in rodent cells by mild cold stress (32 degrees C) . Cirp consists of an N-terminal RNA-binding domain and a C-terminal Gly-rich domain, and plays an essential role in cold-induced suppression of cell proliferation . We report here the cloning of a cDNA encoding an 18-kDa protein with 95.3% identity in an amino-acid sequence to that of mouse Cirp . The human CIRP gene has been mapped to the chromosomal locus 19p13.3 by fluorescence in-situ hybridization . CIRP mRNA is constitutively expressed in all cell lines examined, including K562, HepG2, NC65, HeLa, T24, and NEC8 cells . In all of them, the levels of CIRP mRNA and protein were increased within 12 h after a temperature down-shift from 37 degrees C to 32 degrees C . These results demonstrated that CIRP is a cold-shock protein in human cells . Identification of CIRP may provide a clue to the regulatory mechanisms of cold responses in human cells. Biochim Biophys Acta, 1997 Dec 12, 1359(3), 187 - 99 In search of differentially expressed genes and proteins; Vietor I et al.; A great challenge for modern cell biology is the successful examination of the co-expression of thousands of genes under physiological or pathological conditions and how the expression patterns define the different states of a single cell, tissue or a microorganism . Gene expression can be analyzed today on a large scale by advanced technical approaches for differential screening of proteins and mRNAs . The identification of differentially expressed mRNAs has been successfully applied to understand gene function and the underlying molecular mechanism(-s) of differentiation, development and disease state . Analysis of gene expression by the systematic mapping of thousands of proteins present in a cell or tissue can be achieved by the use of two-dimensional (2D) gel electrophoresis, quantitative computer image analysis, and protein identification techniques . In this article, we comment on some of these techniques and try to stress their advantages and drawbacks . We show how data from RNA/DNA mapping, sequence information from genome projects and protein pattern profiling can be linked with each other and annotated . These comprehensive approaches permit the study of differential gene and protein expressions in cells or tissues. J Can Dent Assoc, 1997 Dec, 63(11), 837 - 45 Oral scurvy and periodontal disease; Touyz LZ; Although gingival bleeding is a manifestation of both scurvy and inflammatory periodontal disease, the two conditions are distinctly separate entities . The defective collagen synthesis associated with scurvy also manifests many of the same symptoms as deficient vitamin C physiology, but neither condition is associated with periodontal disease . Unlike scurvy, the various periodontal diseases are caused by oral plaque microorganisms . The body's reaction to these microorganisms is strongly influenced by the compromised functioning of leucocytes and monocytes . Although certain infections and systemic diseases cause gingival bleeding, avitaminosis-C does not cause commonly encountered periodontitis . Vitamin C should not be used for the prophylaxis or cure of periodontal disease in otherwise healthy, well-nourished individuals . A patient with bleeding gingivae warrants referral to a periodontist, oral medicine specialist, or appropriately qualified dentist for examination and treatment. Blood, 1998 Jan 15, 91(2), 649 - 55 Acute regulation of glucose transport after activation of human peripheral blood neutrophils by phorbol myristate acetate, fMLP, and granulocyte-macrophage colony-stimulating factor; Tan AS et al.; Activation of human peripheral blood neutrophils by pathogens or by phorbol myristate acetate (PMA), fMLP, or myeloid growth factors generates a respiratory burst in which superoxide production plays an important role in killing invading microorganisms . Although the increased energy demands of activated neutrophils would be expected to be associated with increased glucose uptake and utilization, previous studies have shown that PMA inhibits 2-deoxyglucose (2-DOG) uptake . In this study, we show that PMA activation of neutrophils, isolated by methods not involving hypotonic lysis, increases the rate of 2-DOG uptake and results in a 1.6-fold to 2.1-fold increase in transporter affinity for glucose without changing Vmax . Increased transporter affinity in response to PMA was also observed with 3-O-methyglucose, which is not phosphorylated, and inclusion of glucose in the activation medium further increased respiratory burst activity . Increased 2-DOG uptake and increased transporter affinity for glucose were also observed with the peptide activator, fMLP, and with granulocyte-macrophage colony-stimulating factor (GM-CSF) . The protein kinase C (PKC) inhibitor, calphostin C, and the tyrosine kinase inhibitor, genistein, inhibited both PMA- and fMLP-stimulated 2-DOG uptake . In contrast, genistein inhibited fMLP-induced superoxide production, but had little effect on the PMA-induced response, while staurosporine differentially inhibited PMA-induced superoxide production . These results show that neutrophil activation involves increased glucose transport and intrinsic activation of glucose transporter molecules . Both tyrosine kinases and PKC are implicated in the activation process. Curr Opin Biotechnol, 1997 Dec, 8(6), 708 - 12 Will natural products remain an important source of drug research for the future? Nisbet LJ, Moore M. In the highly competitive environment of contemporary pharmaceutical research, natural products provide a unique element of molecular diversity and biological functionality which is indispensable for drug discovery . The emergence of strategies to deliver drug leads from natural products within the same time frame as synthetic chemical screening has eliminated a major limitation of the past . At a more functional level, the application of molecular genetics techniques has permitted the manipulation of biosynthetic pathways for the generation of novel chemical species as well as rendering hitherto uncultivatable microorganisms accessible for secondary metabolite generation . These developments augur well for an industry confronted with the challenge of finding lead compounds directed at the plethora of new targets arising from genomics projects . The exploitation of structural chemical databases comprising a wide variety of chemotypes, in conjunction with databases on target genes and proteins, will facilitate the creation of new chemical entities through computational molecular modelling for pharmacological evaluation. Am Ind Hyg Assoc J, 1997 Dec, 58(12), 876 - 80 Performance of N95 respirators: reaerosolization of bacteria and solid particles; Qian Y et al.; If a respirator does not contain an exhalation value, and the respirator wearer sneezes or coughs, one may expect previously collected particles to be reaerosolized . This may be of special concern in environments contaminated with airborne microorganisms . The percentages of reaerosolization were measured in a test setup where the number of reaerosolized particles were registered by dynamic aerosol size spectrometry relative to the number of previously collected particles or bacteria . Experiments at low relative humidity have shown that the reaerosolization of particles below 1 micron, including Mycobacterium tuberculosis surrogate bacteria, does not exceed 0.025%, even if the re-entrainment air velocity is as high as 300 cm/sec (i.e., 37 times the air velocity through the respirator during breathing under heavy workload conditions) . The reaerosolization of larger particles into dry air was significant at the highest re-entrainment velocity of 300 cm/sec, which simulates violent sneezing or coughing: 0.1% for 3 microns and about 6% for 5-micron test particles . No reaerosolization was detected at relative humidity levels exceeding 35% at these conditions . Thus, it is concluded that the reaerosolization of particles and bacteria, collected on the fibrous filters of N95 respirators, is insignificant at conditions encountered in respirator wear. J Biol Chem, 1997 Dec 5, 272(49), 30672 - 7 Mercury-induced conformational changes and identification of conserved surface loops in plasma membrane aquaporins from higher plants . Topology of PMIP31 from Beta vulgaris L; Barone LM et al.; Aquaporins are integral membrane proteins occurring in mammals, plants, and microorganisms, which serve as channels that permit the bidirectional passage of water through cellular membranes . Higher plants contain abundant levels of aquaporins in both the tonoplast and plasma membrane . Aquaporins contain six transmembrane segments with three surface loops located at the apoplastic face of the membrane and two loops at the cytosolic side . In this study, we probed the topology of plasma membrane aquaporins to determine the effects of divalent cations on aquaporin conformation, and to identify structural features that distinguish plasma membrane intrinsic proteins from tonoplast intrinsic proteins . Plasma membrane vesicles from storage tissue of Beta vulgaris L . were subjected to limited proteolysis, and proteolytic fragmentation patterns were detected using affinity-purified antibodies recognizing aquaporins of 31-kDa . In its native membrane-associated state, the 31-aquaporin band, PMIP31, was refractory to proteolysis by trypsin . However, mercuric compounds specifically induced a conformational change resulting in the exposure of a proteolytic cleavage site and formation of a unique 22-kDa proteolytic fragment (p22) . N-terminal sequence analysis of p22 established its identity as an aquaporin-derived fragment . Topological studies using sealed right-side-out plasma membrane vesicles established that the proteolytic cleavage site is located at surface loop C, the second apoplastic loop, immediately preceding the sequence Gly-Gly-Gly-Ala-Asn . The Gly-Gly-Gly-Ala-Asn-X-X-X-X-Gly-Tyr motif of loop C and a 14 amino acid motif in apoplastic loop E, Thr-Gly-Ile/Thr-Asn-Pro-Ala-Arg-Ser-Leu/Phe-Gly-Ala-Ala-Ile/Val-Ile/ Val-Phe/Tyr-Asn are completely conserved in all known higher plant aquaporins of plasma membrane origin and are not present in any of the known tonoplast intrinsic proteins . These results demonstrate that the two highly conserved plasma membrane intrinsic protein surface loops are structural features that clearly distinguish plasma membrane from tonoplast aquaporins. J Telemed Telecare, 1995, 1(4), 229 - 33 Remote interpretation of microbiology specimens based on transmitted still images; Akselsen S et al.; A pilot study was carried out to examine the feasibility of the remote interpretation of microbiology specimens, that is, micro-organisms grown on agar in Petri dishes . The objective of our study was to decide whether still images contained enough information for microbiology specialists to identify the microorganisms accurately . A representative sample of microbiology specimens grown on the most commonly used agar media was used . Still images were captured using a video camera and a PC-based system . The results from a pilot study with the first video camera were discouraging, the interpretations differing in five out of 22 specimens; results with a second video camera were also disappointing . Images were then captured on photographic film, at a considerably higher resolution than images captured by the digitizer board in the PC . Again, however, the results were disappointing . We conclude that interpretation of microbiology specimens based exclusively on visual information is problematical . Remote microbiology interpretation in the future will require images of higher information content (e.g., including three-dimensional information), and will probably require additional information as well, from other senses, such as smell. J Antimicrob Chemother, 1997 Nov, 40(5), 622 - 30 Antibiotic susceptibilities of mycoplasmas and treatment of mycoplasmal infections; Taylor-Robinson D et al.; Mycoplasmas are the smallest free-living microorganisms, being about 300 nm in diameter . They are bounded by a triple-layered membrane and, unlike conventional bacteria, do not have a rigid cell wall . Hence, they are not susceptible to penicillins and other antibiotics that act on this structure . They are, however, susceptible to a variety of other broad-spectrum antibiotics, most of which only inhibit their multiplication and do not kill them . The tetracyclines have always been in the forefront of antibiotic usage, particularly for genital tract infections, but macrolides are also widely used for respiratory tract infections . Indeed, in comparison with the tetracyclines, erythromycin, the newer macrolides, the ketolides and the newer quinolones have equal or sometimes greater activity . The two latter antibiotic groups also have some cidal activity . The antibiotic susceptibility profiles of several mycoplasmas of human origin are presented, those of Mycoplasma pneumoniae and Mycoplasma genitalium being similar . Apart from the penicillins, mycoplasmas are innately resistant to some other antibiotics, for example the rifampicins . In addition, some may develop resistance, either by gene mutation or by acquisition of a resistance gene, to antibiotics to which they are usually sensitive . Resistance of mycoplasmas to tetracyclines is common and due to acquisition of the tetM gene . The antibiotic susceptibility pattern may be influenced greatly by the source of the mycoplasma; for example, one recovered from a contaminated eukaryotic cell culture that has been subjected to extensive antibiotic treatment may have an antibiotic profile quite different from the same mycoplasmal species that has been recovered directly from a human or animal source . Mycoplasmas may be difficult to eradicate from human or animal hosts or from cell cultures by antibiotic treatment because of resistance to the antibiotic, or because it lacks cidal activity, or because there is invasion of eukaryotic cells by some mycoplasmas . Eradication may be particularly difficult in immunosuppressed or immunodeficient individuals, particularly those who are hypogammaglobulinaemic . The regimes that are most likely to be effective in the treatment of respiratory or genitourinary mycoplasmal infections are presented. Electrophoresis, 1997 Oct, 18(11), 1909 - 11 Enhancement of resolution of low molecular weight RNA profiles by staircase electrophoresis; Cruz-Sanchez JM et al.; Stable low molecular weight (LMW) RNA comprises molecules used in the taxonomy of microorganisms and in studies on the microbial diversity of populations . However, the use of electrophoretic techniques has been hampered due to the low resolution obtained with techniques used for the separation of this kind of molecule . In this work we develop an electrophoretic method (staircase electrophoresis) that increases the resolution of the technique . This improvement in the resolution adequately resolves the three zones that integrate the profiles of LMW RNA: ribosomic 5S RNA (5S rRNA), class 2 transfer RNA (tRNA), and class 1 transfer RNA, allowing the technique to be applied to taxonomic studies (diagnostic, the identification of the individuals), phylogenetic studies and studies on naturally occurring microbial populations. J Infect Dis, 1998 Jan, 177(1), 81 - 7 Complement activation in patients with sepsis is in part mediated by C-reactive protein; Wolbink GJ et al.; The involvement of C-reactive protein (CRP) in the activation of complement in patients with sepsis was investigated . In 104 patients with infections of varying severity, circulating levels of CRP-complement complexes, which are specific indicators for CRP-mediated complement activation, were assessed . Complement-CRP complexes were increased in almost all patients and correlated significantly with levels of C3a (r = .59; P < .001) and C-reactive protein (r = .76; P < .001) . In addition, they correlated with levels of secretory phospholipase A2 (r = .59; P < .001) . Levels of complement-CRP complexes in patients with a pneumococcal type of infection were similar to those in patients with other types of infections . Complement-CRP complexes were significantly higher in patients with shock (P = .01) and in patients who died (P = .03) . These results demonstrate that part of the complement activation in patients with sepsis is independent from a direct interaction with microorganisms but rather results from an endogenous mechanism involving CRP. Mycoses, 1997, 40 Suppl 1, 17 - 21 {Dermatophytes do not produce sialidase in vitro}; Krug G et al.; Sialidase (EC 3.2.1.18) is a pathogenicity factor of many microorganisms, and may also play a role in adhesion of dermatophytes to the epithelia of their hosts by the hydrolytical cleavage of terminal, negatively charged sialic acids of glycoconjugates on the cell surfaces, thus allowing fungal lectins to bind to the subterminal sugars . Therefore, 116 strains of seven species of dermatophytes were investigated for sialidase production . Two highly sensitive, quantitative sialidase assays were applied to cell homogenates and culture supernatants from seven different media of the fungi, but were always negative for sialidase activity . However, sialidase activity was always detected in Ophiostoma stenoceras used as a positive control cultivated in parallel; the enzyme was inducible by sialylated mucins . A sialidase-dependent pathomechanism for dermatophytes appears unlikely based on the results presented. J Gerontol Nurs, 1997 Nov, 23(11), 11 - 6; quiz 54 Occupational health and the the provision of nursing care for older adults; Rymer TE; Providing nursing care to the senior citizen is a rewarding and exciting area of nursing . Care is provided in a vast array of settings from home health and day care to extended residential care sites . These challenges require health care workers to be adaptive and innovative, attentive to detail, compassionate teachers, and promoters of safety for their patients and for themselves . An understanding of the transmission of microorganisms, knowledge of protective mechanisms, and a focus on safety will help ensure good health for the nurse and the elder client . Maintenance of occupational safety and health are combined efforts for the employee and the employer working together to ensure a safe environment for those who need services and those who provide them. Intensive Care Med, 1997 Oct, 23(10), 1024 - 32 Facial mask noninvasive mechanical ventilation reduces the incidence of nosocomial pneumonia . A prospective epidemiological survey from a single ICU; Guerin C et al.; OBJECTIVE: To evaluate the impact of noninvasive positive pressure mechanical ventilation (NPPV) on ventilator-associated pneumonia (VAP) . DESIGN: Prospective observational study . SETTING: Medical intensive care unit (ICU) of a university teaching hospital . PATIENTS: Cohort of 320 consecutive patients staying in the ICU more than 2 days and mechanically ventilated for > or = 1 day . MEASUREMENTS AND RESULTS: VAP was diagnosed when, satisfying classical clinical and radiological criteria, fiberoptic bronchoalveolar lavage and/or protected specimen brush grew > or = 10(4) and > or = 10(3) CFU/ml, respectively, of at least one microorganism . Patients were classified into four subgroups according to the way in which mechanical ventilation was delivered: NPPV then tracheal intubation (TI) (n = 38), TI then NPPV (n = 23), TI only (n = 199), and NPPV only (n = 60) . Occurrence of VAP was estimated by incidence rate and density of incidence . Risk factors for VAP were assessed by logistic regression analysis . Twenty-seven patients had 28 episodes of VAP . The incidence rates for patients with VAP were 18% in NPPV-TI, 22% in TI-NPPV, 8% in TI, and 0% in NPPV (p < 0.0001) . The density of incidence of VAP was 0.85 per 100 days of TI and 0.16 per 100 days of NPPV (p = 0.04) . Logistic regression showed that length of ICU stay and ventilatory support were associated with VAP . CONCLUSIONS: There is a significantly lower incidence of VAP associated with NPPV compared to tracheal intubation . This is mainly explained by differences in patient severity and risk exposure. Am J Respir Crit Care Med, 1997 Dec, 156(6), 1908 - 14 Community-acquired pneumonia in the elderly . Clinical and nutritional aspects; Riquelme R et al.; Community-acquired pneumonia (CAP) in the elderly has a different clinical presentation than CAP in other age groups . Confusion, alteration of functional physical capacity, and decompensation of underlying illnesses may appear as unique manifestations . Malnutrition is also an associated feature of CAP in this population . We undertook a study to assess the clinical and nutritional aspects of CAP requiring hospitalization in elderly patients (over 65 yr of age) . One hundred and one patients with pneumonia, consecutively admitted to a 1,000-bed teaching hospital over an 8-mo period, were studied (age: 78 +/- 8 yr, mean +/- SD) . Nutritional aspects and the mental status of patients with pneumonia were compared with those of a control population (n = 101) matched for gender, age, and date of hospitalization . The main symptoms were dyspnea (n = 71), cough (n = 67), and fever (n = 64) . The association of these symptoms with CAP was observed in only 32 patients . The most common associated conditions were cardiac disease (n = 38) and chronic obstructive pulmonary disease (COPD) (n = 30) . Seventy-seven (76%) episodes of pneumonia were clinically classified as typical and 24 as atypical . There was no association between the type of isolated microorganism and the clinical presentation of CAP, except for pleuritic chest pain, which was more common in pneumonia episodes caused by classical microorganisms (p = 0.02) . This was confirmed by a multivariate analysis (relative risk {RR} = 11; 95% confidence interval {CI}: 1.7 to 65; p = 0.0099) . The prevalence of chronic dementia was similar in the pneumonia cohort (n = 25) and control group (n = 18) (p = 0.22) . However, delirium or acute confusion were significantly more frequent in the pneumonia cohort than in controls (45 versus 29 episodes; p = 0.019) . Only 16 patients with pneumonia were considered to be well nourished, as compared with 47 control patients (p = 0.001) . Kwashiorkor-like malnutrition was the predominant type of malnutrition (n = 65; 70%) in the pneumonia patients as compared with the control patients (n = 31; 31%) (p = 0.001) . The observed mortality was 26% (n = 26) . Pleuritic chest pain is the only clinical symptom that can guide an empiric therapeutic strategy in CAP (typical versus atypical pneumonia) . Both delirium and malnutrition were very common clinical manifestations of CAP in our study population. Antibiot Khimioter, 1997, 42(9), 3 - 13 {Determination of the nucleotide sequence of the silent aminoglycoside phosphotransferase gene in Streptomyces rimosus P3}; Akopiants KE et al.; The nucleotide sequence BgIII-Pstl of the DNA fragment containing the aph gene cloned from Streptomyces rimosus P3 not producing aminoglycoside antibiotics was determined . The aph gene was shown to encode neomycin phosphotransferase differing by the substrate specificity from the enzyme encoded by the aph genes from the organisms producing aminoglycoside antibiotics . Comparison of the cloned aph VIII gene and its product with the aph genes and their products from the antibiotic-producing actinomycetes and clinical microbial strains permitted to consider it as a representative of the third group genes which are likely widely distributed in soil microorganisms not producing aminoglycoside antibiotics . The gene length was 777 nucleotide pairs at the average GC content of 67 per cent . Comparison of the nucleotide and protein sequences of the S.rimosus gene with those of the genes cloned from the aminoglycoside-producing organisms (S.fradiae and Micromonospora chalcea) revealed high homology in the 3'-end region of the genes . However, the homology percentage by DNA for the S.rimosus gene was lower than that for the genes from the other organisms on their comparison with each other: 56-67 and 82-86 per cent respectively . Comparison of the profiles of the protein hydrophilic properties revealed differences in the region of the first 110 amino acids of the gene under the investigation . The amino acid sequence contained all the highly conservative regions characteristics of aminoglycoside phosphotransferases but had several amino acid substitutes detected for the first time including those in the region responsible for the antibiotic binding. Rev Argent Microbiol, 1997 Jul-Sep, 29(3), 157 - 66 {Mycoplasmas and AIDS}; Coronato S; AIDS is a complex illness due to HIV type 1 and 2 infection . It is characterized by an important immunodeficiency mainly caused by depletion of CD4+ T lymphocytes . The reasons for this depletion have not been sufficiently clarified yet . In 1986, Shy Ching Lo astonished the scientific community with reported evidence concerning the direct role played by mycoplasma in the etiopathology of AIDS . Since then, different theories have pointed to mycoplasma as cofactors, commensals or opportunistic agents . Although in vivo and in vitro experiments are controversial they suggest a possible mechanism that would explain the synergism between both agents: the mycoplasma belonging to normal intestinal flora could move to urethra, oropharynx or blood due to high risk sexual practice . There it would proliferate favoured by early immunological disorders related to HIV . It has been speculated that several microorganisms including mycoplasma, acting as superantigens, could induce a chronic CD4+ and CD8+ T lymphocytes activation resulting in apoptosis of the infected lymphocytes . The release of cytokines induced by mycoplasma could influence the progression of the disease. Dtsch Med Wochenschr, 1997 Nov 7, 122(45), 1377 - 81 {Chlamydia pneumoniae infection in a family}; Freidank HM et al.; HISTORY AND SYMPTOMS: For 11 weeks a 38-year-old woman had suffered from a respiratory infection with peribronchitis, nocturnal coughing fits and earache . INVESTIGATIONS, TREATMENT AND COURSE: The Chlamydia-CFR titre was raised . Subsequent throat swabs of her husband and two daughters grew Chlamydia pneumoniae (C.p.), but not in her case; 5 days earlier she had been started on roxithromycin . 3 weeks before the patient fell ill her two daughters had a flu-like illness with cough and subfebrile temperature and her husband also had flu-like symptoms, which regressed after few days . Antibiotic treatment with roxithromycin improved the symptoms in the mother and older daughter, but the younger daughter was not given treatment because she had no symptoms at the time the microorganism had been isolated . She developed joint symptoms, like those of reactive arthritis, 12 weeks after the mother's illness had begun, and conjunctivitis 5 weeks later . CONCLUSIONS: It is likely that the daughters had the C.p . infection first and then infected their parents . While the father's and older daughter's illness quickly regressed, the mother became quite ill . Her serology was positive for a primary infection in adulthood, but in the daughters the serology was negative and, despite demonstration of the organism, the diagnosis of acute C.p . infection could not be made. Enferm Infecc Microbiol Clin, 1997 Aug-Sep, 15(7), 373 - 7 {Intensive care for patients with the human immunodeficiency virus}; Esteban J et al.; BACKGROUND: HIV infection is of special importance in Spain, occasionally requiring the use of intensive care units (ICU) in the management of these cases . METHODS: A retrospective review of the seropositive HIV patients was carried out of those requiring ICU admission in the authors' center over a three year period . Twenty-five patients were included, with the cause for admission into ICU, the clinical features, mortality and prognostic factors being reported . RESULTS: Most of the patients presented as a risk behavior by intravenous drug addiction and did present AIDS prior to hospitalization . The most frequent cause for admission in the ICU was respiratory failure produced by germs other than Pneumocytis carinii, placing this microorganism in second place . The mean time of admission in the ICU was 10.8 days, with 88% requiring mechanical ventilation and 56% the use of vasoactive drugs . The ICU mortality was 48% . No survivor died posteriorly on the ward . Factors of bad prognosis were: less than 200 CD4 lymphocytes/mm3, low albumin levels and high creatinine in addition to the need for vasoactive drugs with no evidence of other data presenting statistical significance . Fourty-four percent of the survivors remain alive at 12 months of hospital discharge . CONCLUSIONS: Respiratory failure produced by Pneumocystis carinii or by other germs is the most frequent cause for ICU admission of patients with HIV infection . Renal failure, lymphopenia, and hypoalbuminemia are predictors of mortality in almost 50% of the cases. Zentralbl Hyg Umweltmed, 1996 Dec, 199(2-4), 334 - 49 {Procedures for hand hygiene in German-speaking countries}; Rotter M; According to the field of application, strategies for the prevention of the transfer of microbial skin flora from the hands must consider the various categories of flora: transient, resident or stemming from infected lesions on the hands (infection flora) . Depending on the species and virulence of the microorganism and of the susceptibility of the infection target, transient flora may or may not be of pathogenic importance . In contrast, resident skin flora is usually regarded as pathogenic only under certain circumstances such as in surgery, especially with transplantation of foreign bodies and in highly susceptible hosts . Microorganisms stemming from infected lesions are of proven pathogenicity . In the non-surgical field, only the transient and infection flora from the hands play a role . Such lesions are an absolute contraindication for patient-care, preparation of pharmaceuticals or foodstuff . In some procedures, the transmission of transient flora can be prevented by use of the non-touch technique ("instruments instead of fingers") or by the intelligent use of protective gloves . Hands already contaminated may be rendered safe by procedures for the elimination of transients such as handwashing, hygienic handwash and hygienic hand rub (in the order of increasing efficacy) . Among all useable chemicals, ethanol, isopropanol and n-propanol (in the order of increasing efficacy) are the strongest and fastest agents . Furthermore, the duration of treatment (between 30 and 60 s) significantly influences the achievable reduction of microbial release . According to the new European standards (CEN) for testing chemical disinfectants and antiseptics, products for hygienic handwash must be significantly more efficacious than unmedicated soap, on artificially contaminated hands . In contrast, products for the hygienic hand rub must not be significantly less efficacious than a reference disinfection including isopropanol 60% vol rubbed onto the hands of the same volunteers during 1 min . By this, the average reduction of microbial release amounts to 4.2 to 4.4 lg, in our hands . The effectiveness of procedures for the hygienic handwash is usually significantly lower than that of alcoholic rubs . Therefore, in hospitals, they can be used only in certain indications such as patient care in reverse isolation, preparation of pharmaceuticals or foodstuff . In the surgical field, where not only transient but also resident flora is a cause of post-operative infection, the microbial release from the hands of the surgical team into the surgical wound must be prevented by using surgical gloves . Because of frequent glove lesions, a surgical hand disinfection is usually performed before donning gloves to keep a possible inoculumn as small as possible . Also in this field of application, alcoholic rubs proved to be significantly more effective than washing hands with antiseptic detergents . There exists a strong positive correlation of the reduction of microbial release and the duration of hand treatment, between 1 and 5 min . The European test standards (CEN) require products for surgical hand disinfection to be at least as efficacious as a reference disinfection of clean hands, which are constantly rubbed and kept wet with n-propanol 60% vol during 3 min . By this, the achievable average reduction of the microbial release ranges between 2.0 and 2.4 lg . In contrast, antiseptic washing procedures with preparations containing povidone-iodine, chlorhexidine gluconate or triclosan reduce the bacterial release within 2-5 min only by 0.5 to 1.2 lg . Some of them exert a bacteriostatic sustaining effect which is not found with alcoholic preparations . This, however, is not necessary with the latter as the initial bacterial reduction is that strong that restitution of the skin flora takes > 3 hours . Alcoholic preparations are at least as tolerable for the skin as antiseptic detergents, if not better, if they contain suitable emollients . Because dilution renders alcohols i Zentralbl Hyg Umweltmed, 1995 Oct, 198(1), 72 - 83 {Hygienic processing of dental transmission instruments (handpieces and instruments, turbines) in dental practice}; Graf W et al.; The efficacy of a steam disinfection method for the processing of used instruments in dental practice was tested with 274 high-speed turbines and handpieces . The instruments treated with streaming steam were both taken out of the dental practice of treatment and artificially contaminated with five different test-microorganisms (E . coli, P . aeruginosa, S . aureus, E . faecium, C . albicans) and different concentrations of serum (1%, 10%, 20%) . In spite of the narrow channels of these instruments a microbial reduction of more than 5 log10 steps could be achieved in all cases . Thus dental high-speed turbines and handpieces can reliably be disinfected with streaming steam. Gene, 1997 Nov 12, 201(1-2), 31 - 6 Evidence for two types of subunits in the bacterioferritin of Magnetospirillum magnetotacticum; Bertani LE et al.; In order to investigate the role of bacterioferritin (Bfr) in the biomineralization of magnetite by microorganisms, we have cloned and sequenced the bfr genes from M . magnetotacticum . The organism has two bfr genes that overlap by one nucleotide . Both encode putative protein products of 18 kDa, the expected size for Bfr subunits, and show a strong similarity to other Bfr subunit proteins . By scanning the DNA sequence databases, we found that a limited number of other organisms, including N . gonorrhea, P . aeruginosa, and Synechocystis PCC6803, also have two bfr genes . When the sequences of a number of microbial Bfrs are compared with each other, they fall into two distinct types with the organisms mentioned above having one of each type . Differences in heme- and metal-binding sites and ferroxidase activities of the two types of subunits are discussed. Scand J Rheumatol, 1997, 26(5), 389 - 91 SLE with death from acute massive pulmonary hemorrhage caused by disseminated strongyloidiasis; Setoyama M et al.; We describe a case of disseminated strongyloidiasis involving a female patient with active systemic erythematosus (SLE) . The cause of death was massive pulmonary hemorrhage induced by a filariform larvae infection . This was initially diagnosed during examination of the bronchoalveolar lavage fluid just 2 days before her death . The case indicated that immunosuppressed individuals would be paid attention to possible parasitic infection prior to starting therapy even in non-endemic areas as well as other microorganisms. FEBS Lett, 1997 Oct 27, 416(3), 302 - 6 A novel small stable RNA, 6Sa RNA, from the cyanobacterium Synechococcus sp . strain PCC6301; Watanabe T et al.; We isolated a novel RNA species from the unicellular cyanobacterium Synechococcus PCC6301 and determined its gene sequence . This novel RNA was termed 6Sa RNA from its length (185 nt) . Cross-hybridization of 6Sa RNA to other related microorganisms suggests that its existence is restricted to the Synechococcus genus or related organisms . A high level of accumulation of this RNA was observed by Northern analysis, indicating that 6Sa RNA is stable in cells . Computer-aided prediction of the 6Sa RNA secondary structure also supports its stability. Curr Opin Hematol, 1996 Jan, 3(1), 94 - 9 Apoptosis of neutrophils; Homburg CH et al.; Neutrophils are the most abundant leukocytes and serve as a first line of defense against infectious microorganisms . For this purpose, neutrophils contain granules filled with proteolytic and other cytotoxic enzymes . Neutrophils have the shortest lifespan of all leukocytes . To prevent senescent neutrophils from releasing their toxic contents into the surrounding tissues, these cells become apoptotic and are then internalized by tissue macrophages . Recent studies have revealed more details about effects of cytokines on neutrophil apoptosis and on the uptake of apoptotic neutrophils by macrophages . In addition, the intracellular events leading to apoptosis are slowly being unraveled. Curr Opin Hematol, 1996 Jan, 3(1), 19 - 26 Peptide transport in antigen presentation; Marusina K et al.; The TAP1/2 complex translocates peptides from the cytosol into the endoplasmic reticulum . In the rat, TAP polymorphism affects the pool of peptides presented by major histocompatibility complex class I molecules, whereas in mouse and humans the functional consequences of observed structural polymorphism have not yet been determined . Peptide binding to TAP precedes ATP binding, and ATP hydrolysis is required to release and translocate peptides . Cytosolic peptides entering the class I pathway via the TAP complex may differ significantly in size and sequence . Not all peptides that are able to bind to TAP can be translocated into the endoplasmic reticulum, and TAP-binding affinity does not directly correlate with antigenicity of the peptide . Most translocated peptides are released from the endoplasmic reticulum by an incompletely defined ATP-dependent mechanism . TAP interaction with class I molecules stimulates peptide binding and transport by TAP and may also facilitate efficient loading of class I with peptides . Pathogenic microorganisms, such as herpes simplex virus, may encode inhibitors of TAP-mediated peptide transport in order to evade immune surveillance. Mol Biotechnol, 1997 Oct, 8(2), 139 - 45 Identification of microorganisms using random primed PCR; Mileham AJ; The polymerase chain reaction has facilitated the use of molecular approaches in microbiology including new strategies for the rapid identification of micro-organisms . Approaches based on the use of random primers and standard conditions, allows characteristic DNA fingerprints to be generated from any micro-organism even in the absence of information about its DNA sequence . Different primers can be used to produce genus-specific, species-specific, or even strain-specific DNA fingerprints . This article covers the background to this strategy, describes three different approaches to generating DNA fingerprints using random primers, and provides experimental detail for one method, RAPD. Mol Biotechnol, 1997 Oct, 8(2), 123 - 34 Overview of vaccines; Ada G; This article lists the vaccines current available for the control of both viral and bacterial infections . They may be attenuated live or inactivated whole microorganisms, or subunit preparations . Many more are in the pipeline and increasing attention is being given to establishing their safety before registration . Following the earlier eradication of smallpox, good progress is now being made toward the global eradication of poliomyelitis and a new program to eliminate measles from the Americas has begun . A variety of new approaches to vaccine development is now available . The hepatitis B virus surface antigen, made by DNA-transfected yeast or mammalian cells, is the basis of the first genetically engineered vaccine . Early in the 21st century, new vaccines based on oligopeptides, recombinant live viral or bacterial vectors (often existing live vaccines), or recombinant DNA plasmids are likely to be registered for human use . The efficacy of vaccines depends on the immune responses generated, and the recent substantial increase in our understanding of the mammalian immune system now offers great opportunities for manipulation to best obtain desired responses . These include mixing vaccine formulations to maximize immune responses, and combining vaccines to simplify their administration . Despite these advances, some persisting infections, such as those caused by HIV, plasmodia, and mycobacteria, still pose a great challenge to vaccine developers. Med Microbiol Immunol (Berl), 1997 Oct, 186(2-3), 75 - 81 Leishmania major infection in C57BL/10 mice differing at the Lps locus: a new non-healing phenotype; Muller I et al.; The course of cutaneous leishmaniasis was examined in mice from two genetically closely related strains, C57BL/10ScCr (Cr) and C57BL/10ScSn (Sn) . Sn mice are able to heal Leishmania major infections, while Cr mice are unable to heal . The cutaneous lesions of the Cr mice progressed continuously and the increase in lesion size was paralleled by an unrestricted growth of the parasites in vivo . Cr mice, in contrast to their Sn counterparts, are highly resistant to all effects of lipopolysaccharide (LPS) . The nonhealing L . major infection in Cr mice is in sharp contrast to the course of infection in another endotoxin-nonresponder mouse strain, C3H/HeJ, which heal infections with L . major . Cr mice exhibit, in addition to the defective LPS responsiveness, an impaired interferon-gamma (IFN-gamma) response after infection with a variety of microorganisms . The insufficient activation of parasitized macrophages to kill intracellular L . major could be due to the inability of splenocytes from infected Cr mice to secrete IFN-gamma upon restimulation with L . major . IFN-gamma is essential for the efficient activation of parasitized macrophages to kill intracellular L . major by producing nitric oxide (NO) . Although bone marrow-derived Cr macrophage do not produce NO in response to LPS, both Sn and Cr macrophages release NO upon stimulation with IFN-gamma and tumor necrosis factor, indicating that they are responsive to activation by these cytokines. Dermatology, 1997, 195 Suppl 2, 42 - 8 Effects of Betaisodona on parameters of host defense; Konig B et al.; The numbers of patients in intensive care units, with immunosuppression, and of elderly people increase in parallel with antibiotic-resistant microorganisms . Therefore the demand for an effective antisepsis increases . Moreover, it became evident that the pathophysiology and the outcome of infection are dependent on the properties of the microorganisms, e.g . synthesis of endo- and exotoxins, and on the host defense, the immune system . In addition to the microbicidal action, we studied the effects of povidone-iodine (PVP-I, Betaisodona) on the generation, release and activity of exotoxins (alpha-hemolysin, phospholipase C, lipase), as well as on granulocyte-derived tissue-destructive enzymes (elastase, beta-glucuronidase) and microbial-induced cytokine generation from human neutrophils . Our results clearly show that PVP-I does not only kill a wide range of bacteria but also inhibits the generation and release of bacterial exotoxins; furthermore, it also inactivates bacterial exotoxins as well as granulocyte-derived tissue-destructive enzymes and cytokines . These data support the usefulness and efficacy of PVP-I as an effective therapeutic agent to combat infection. Hum Genet, 1997 Dec, 101(2), 205 - 7 Assignment of human plasma methylumbelliferyl-tetra-N-acetylchitotetraoside hydrolase or chitinase to chromosome 1q by a linkage study; Eiberg H et al.; Plasma methylumbelliferyl tetra-N-acetylchitotetraoside hydrolase or chitinase (CHIT) might play a role in degrading the chitin wall of some microorganisms . In about 6% of Caucasian people the enzyme shows pseudodeficiency (defined as very low activity without apparent symptoms) . We have mapped this locus by linkage analysis to the marker D1S306 (z = 4.00 at theta M = F = 0.0) on chromosome 1q between the flanking markers D1S191 and D1S245 in the area of 1q31-1qter. Trends Microbiol, 1997 Nov, 5(11), 450 - 4 Filamentous growth in budding yeast; Kron SJ; The recent discovery that some laboratory strains of Saccharomyces cerevisiae are capable of limited filamentous growth has stimulated genetic analysis of dimorphism in this microorganism . The puzzle of why most strains are nonfilamentous is now resolved . Remarkably, a single point mutation with broad consequences separates these domesticated yeast from their wild ancestors. J Leukoc Biol, 1997 Dec, 62(6), 702 - 9 Elimination, blocking, and activation of macrophages: three of a kind? van Rooijen N, Sanders A. In mammals, macrophages are multifunctional cells . Apart from their scavenger role in the clearance of non-self materials such as microorganisms and altered-self materials such as apoptotic cells, senescent erythrocytes, immune complexes, and inflammatory products, they play a crucial role in the regulation of both innate and acquired immunity . Whereas the former activity is based on phagocytosis and intracellular degradation, the latter activity largely depends on the production and secretion of a panel of regulatory molecules such as cytokines, chemokines, and nitrogen oxide (NO) . Depletion of macrophages and blocking of phagocytosis form important approaches to study the role of these cells in various host defense mechanisms . Moreover, the efficacy of drug- and gene-targeting, based on the application of particulate carrier devices, can be improved in this way . However, compounds originally described as efficacious blockers of phagocytosis simultaneously activate their production of cytokines and NO . Moreover, elimination, blocking, as well as activation of macrophages are all dependent on the concentration of such compounds . When administered in vivo, they will reach some macrophages in a high and others in a low concentration . As a consequence, the former cells may be eliminated or blocked, whereas the latter are activated by the same treatment . In this review, the various methods for suppression of macrophage functions are compared and requirements for the development of new, selective, and organ-specific macrophage-suppressing devices are discussed. Immunol Invest, 1997 Aug-Dec, 26(5-7), 631 - 47 CD5+ B cell-dependent regulation of the murine T-cell independent immune response against the human blood group A antigen; Neron S et al.; The CD5+B lymphocyte (B1a) population is known to be involved in most immune responses to microorganism TI antigens . Moreover, xid mice deficient for immune responses against TI-2 antigens are known to lack the B1a population, suggesting a role for B1a cells in TI-2 immune responses . We previously established that the oligosaccharide human blood group A antigen stimulated murine TI-2 immune responses . In this work, we show that the frequency of anti-A-secreting hybridomas was higher in mice with larger splenic B1a populations and that in vivo anti-CD5 treatment reduced anti-A immune response without affecting the response against TD RBC antigens . A similar effect was observed by in vitro anti-CD5 treatment of splenocytes . The in vivo anti-CD5 treatment also interfered with the immunization-dependent increase in splenocyte numbers . These results are in agreement with an important role for the B-cell CD5 receptor in the regulation of TI-2 immune responses possibly mediated by its interaction with the CD72 ligand. Biochem Biophys Res Commun, 1997 Nov 26, 240(3), 669 - 72 Evidence that exposure of particulate air pollutants to human and rat alveolar macrophages leads to differential oxidative response; Rahman Q et al.; Macrophages and inflammatory cells generate active oxygen species in the process of killing and degrading microorganisms . Air pollutant particles may be ingested by macrophages and stimulate the same mechanisms to produce a long term oxidative burden to the lung if particles are not degraded . In the present study human and rat alveolar macrophages (AM) were compared in their response to inhaled particles using luminol dependent chemiluminescence (CL) and peroxide dependent CL assays . Cytotoxicity was measured by the release of lactate dehydrogenase (LDH) activity in the supernatant . Human AM produced more oxidants than rat AM whether, unstimulated, after addition of particles or addition of particles then peroxidase . Human AM also had a different spectrum of response to the same particles . Our results suggest that human macrophages produce more reactive oxygen species in respond to particles than rat AM. Dev Comp Immunol, 1997 Sep-Oct, 21(5), 385 - 95 The prophenoloxidase activating system of the shrimp Penaeus paulensis and associated factors; Perazzolo LM et al.; In the present study we investigated the proPO activating system of the penaeid Penaeus paulensis, focusing on its role in the shrimp immune system . The great majority of PO activity (more than 90%) was found in the shrimp hemocytes . The enzyme activity was greatly enhanced by components of microorganism cell walls, such as LPS and beta-1,3-glucans, suggesting its involvement in non-self recognition . PO activity was also found in the shrimp serum and trypsin, and LPS were able to increase the enzyme activity . Thus, serum can be used as an alternative for the study of the shrimp proPO activating system, as it is much more readily obtained than HLS . PO activity was cation-dependent, and 5 mM of calcium and 10 mM of magnesium were the optimal concentrations for the enzyme activity . An immune factor was found in the shrimp HLS, capable of inducing cell-adhesion and degranulation of the penaeid hemocytes. Am J Obstet Gynecol, 1997 Nov, 177(5), 1230 - 7 Lipopolysaccharide binding protein and soluble CD14 receptor protein in amniotic fluid and cord blood in patients at term; Roos T et al.; OBJECTIVES: Our purpose was to examine whether lipopolysaccharide binding protein and soluble CD14 are present in amniotic fluid and to determine whether the lipopolysaccharide binding protein and soluble CD14 concentrations are associated with indicators of infection or labor at term . A lipopolysaccharide-lipopolysaccharide binding protein complex activates macrophages through soluble CD14 at lipopolysaccharide concentrations up to 100 times lower than required with lipopolysaccharide alone . Thus lipopolysaccharide binding protein and soluble CD14 in amniotic fluid could explain the high concentrations of cytokines found in amniotic fluid of culture-positive patients and may even explain the presence of cytokines in some culture-negative patients . STUDY DESIGN: Healthy women at term undergoing cesarean section had amniotic fluid, chorioamnion, decidua, and cord blood obtained . Lipopolysaccharide binding protein was measured by enzyme-linked immunosorbent assay . Amniotic fluid was cultured and assayed for cytokines, and the chorioamnion and decidua were cultured and examined histologically . RESULTS: Lipopolysaccharide binding protein and soluble CD14 were present in all amniotic fluids and fetal cord blood . An elevated level of lipopolysaccharide binding protein (270 ng/ml/mg of protein) was present in the amniotic fluid of 12 (36%) of the 33 patients . An elevated level was associated with microorganisms in the chorioamnion and decidua, cytokines (tumor necrosis factor-alpha, interleukin-6, and interleukin-8) in amniotic fluid, histologic chorioamnionitis, and labor . Among patients in labor, the concentration of lipopolysaccharide binding protein appeared independent of microorganisms in the amniotic fluid . CONCLUSIONS: Lipopolysaccharide binding protein and soluble CD14 are present in amniotic fluid, and concentrations of lipopolysaccharide binding protein are elevated in patients in labor with and without evidence of infection . Lipopolysaccharide binding protein and soluble CD14 may mediate intrauterine inflammatory responses at term. J Infect Dis, 1997 Dec, 176(6), 1552 - 8 Inhibition of virulent Mycobacterium tuberculosis by Bcg(r) and Bcg(s) macrophages correlates with nitric oxide production; Arias M et al.; The Nramp1 gene controls macrophage resistance or susceptibility to several intracellular microorganisms; however, there is conflicting evidence regarding its role during infection with virulent Mycobacterium tuberculosis . Nitric oxide (NO) is a potent antimycobacterial agent produced by macrophages, which is also regulated by Nramp1 . The in vitro ability of B10R (resistant) and B10S (susceptible) murine macrophages to inhibit M . tuberculosis H37Rv and to produce NO in response to infection and interferon-gamma (IFN-gamma) was compared . Infected B10R macrophages inhibited {3H}uracil incorporation by M . tuberculosis and produced higher amounts of NO than did B10S macrophages . IFN-gamma increased the inhibitory activity of both cells . Inhibition of M . tuberculosis by IFN-gamma-activated B10R macrophages was reversed by N(G)-monomethyl-L-arginine (N(G)MMA) . L-arginine restored NO production and increased the antimycobacterial activity by IFN-gamma-stimulated N(G)MMA-treated macrophages . The Bcg/Nramp1 gene may regulate macrophage resistance or susceptibility to virulent M . tuberculosis by a differential capability of these cells to produce NO. Infect Immun, 1997 Dec, 65(12), 5272 - 8 Effects of mycobacteria on regulation of apoptosis in mononuclear phagocytes; Klingler K et al.; Since apoptosis is observed in tuberculous granulomata, we investigated the molecular mechanisms underlying the apoptotic pathway in an in vitro model of mycobacterial infection of mononuclear phagocytes . We postulated that Mycobacterium tuberculosis could trigger the apoptotic pathway in macrophages, resulting in death of the microorganism by modulating the expression of bcl-2, bax, bcl-xL, and bcl-xS . We found that the mRNA of bcl-2, an inhibitor of apoptosis, was downregulated in peripheral blood monocytes (PBM) between 2 and 6 h following infection with M . bovis BCG or induction with heat-killed M . tuberculosis H37Ra . Western analysis showed a downregulation of the Bcl-2 protein, with a half-life of 24 h . At the same time points, there was no change in the expression of Bax or Bcl-xS, inducers of apoptosis, but Bcl-xL, another inhibitor of apoptosis, was minimally upregulated by BCG . To determine if apoptosis could be a mechanism for growth inhibition in vivo, we obtained alveolar macrophages by bronchoalveolar lavage from involved sites in patients with active pulmonary tuberculosis . Using the TUNEL (terminal deoxynucleotidyltransferase mediated nick end labeling) technique, we observed significantly more apoptosis in involved segments of five tuberculosis patients (14.8 +/- 1.9%) than in those of normal controls (<1%, P = 0.02) or in uninvolved segments (4.3 +/- 0.9%, P < 0.05) . We conclude that apoptosis of mononuclear phagocytes induced by M . tuberculosis occurs in vivo and that in an in vitro model of mycobacterial infection, apoptosis may be mediated by downregulation of Bcl-2. Infect Immun, 1997 Dec, 65(12), 5096 - 102 Virulence characteristics of oral treponemes in a murine model; Kesavalu L et al.; This study was designed to investigate the virulence characteristics of Treponema denticola, T . socranskii, T . pectinovorum, and T . vincentii following challenge infection of mice . These microorganisms induced well-demarcated, dose-dependent, raised subcutaneous (s.c.) abscesses which were similar in time of onset, lesion progression, and duration of healing . Only viable cells were capable of inducing these characteristic s.c . abscesses . Histological examination of the skin lesion 3 and 5 days postinfection revealed abscess formation in the s.c . tissues, and abundant spiral organisms were demonstrated to be present in the abscess . Host resistance modulation by dexamethasone (neutrophil alteration) and cyclophosphamide (neutrophil depletion) pretreatment had a minimal effect on the virulence expression by any of these treponemes . The T . denticola isolates demonstrated significant trypsin-like protease (TLPase) activity, while both T . socranskii and T . vincentii were devoid of this activity . Interestingly, T . pectinovorum strains were heterogeneous with respect to TLPase as high producers, low producers, and nonproducers . However, no differences in lesion formation were noted regardless of whether the species expressed this proteolytic activity or whether treatment with N alpha-p-tosyl-L-lysine chloromethyl ketone (TLCK) and dithiothreitol was performed . These results showed that (i) a murine model may be used to evaluate virulence expression by oral treponemes; (ii) while TLPase activity varies among the oral treponemes, this protease does not appear to participate in abscess induction in the mouse model; and (iii) T . pectinovorum strains show variation in TLPase activity. J Bacteriol, 1997 Dec, 179(23), 7360 - 8 A study of iterative type II polyketide synthases, using bacterial genes cloned from soil DNA: a means to access and use genes from uncultured microorganisms; Seow KT et al.; To examine as randomly as possible the role of the beta-ketoacyl and acyl carrier protein (ACP) components of bacterial type II polyketide synthases (PKSs), homologs of the chain-length-factor (CLF) genes were cloned from the environmental community of microorganisms . With PCR primers derived from conserved regions of known ketosynthase (KSalpha) and ACP genes specifying the formation of 16- to 24-carbon polyketides, two CLF (KSbeta) genes were cloned from unclassified streptomycetes isolated from the soil, and two were cloned from soil DNA without the prior isolation of the parent microorganism . The sequence and deduced product of each gene were distinct from those of known KSbeta genes and, by phylogenetic analysis, belonged to antibiotic-producing PKS gene clusters . Hybrid PKS gene cassettes