Pediatr Infect Dis J, 1997 Feb, 16(2), 222 - 6 Standardization of gastric aspirate technique improves yield in the diagnosis of tuberculosis in children; Pomputius WF 3rd et al.; BACKGROUND: The diagnosis of tuberculous disease in children remains a difficult one, based on epidemiologic investigation, Mantoux skin testing and suggestive radiologic findings . Because children with pulmonary tuberculosis are unable to produce sputum, gastric aspirates remain the procedure of choice for microbiologic confirmation of tuberculous disease; however, yield is frequently low . OBJECTIVES: To evaluate the effect of a standardized gastric aspirate collection protocol on diagnostic culture yield . METHODS: The gastric aspirate culture yield for Mycobacterium tuberculosis in 13 historical control children with clinically confirmed tuberculosis from 1979 to 1994 was compared with the yield in 8 children with tuberculous disease after institution of a standardized gastric aspirate collection protocol involving physician education, strict timing of collection, base neutralization of aspirate specimens and expedited processing . RESULTS: Retrospective survey of gastric aspirate results in Rhode Island from 1979 to 1994 revealed that only 1 of 13 cases (8%) of pediatric pulmonary tuberculosis were confirmed in this manner . During a 12-month period after institution of a protocol, gastric aspirates yielded positive cultures in 4 of 8 children (50%) with pulmonary tuberculosis, a yield that compares favorably with the sensitivities of 20 to 52% published in the literature . CONCLUSIONS: Attention to the technique of gastric aspirate collection, and expedited processing in particular, appears to improve the yield of this diagnostic procedure for pediatric tuberculosis. Appl Environ Microbiol, 1997 Feb, 63(2), 628 - 35 Monocentric and polycentric anaerobic fungi produce structurally related cellulases and xylanases; Li XL et al.; Cellulase and xylanase cDNAs were isolated from a cDNA library of the polycentric anaerobic fungus Orpinomyces sp . strain PC-2 constructed in Escherichia coli . The cellulase cDNA (celB) was 1.8 kb long with an open reading frame (ORF) coding for a polypeptide of 471 amino acids, and the xylanase cDNA (xynA) was 1.2 kb long with an ORF encoding a polypeptide of 362 amino acids . Single transcripts of 1.9 kb for celB and 1.5 kb for xynA were detected in total RNA of Orpinomyces grown on Avicel . Genomic DNA regions coding for CelA and XynA were devoid of introns . The enzymes were highly homologous (80 to 85% identity) to the corresponding enzymes of the monocentric anaerobic fungus Neocallimastix patriciarum and, like those, contained in addition to a catalytic domain, a noncatalytic repeated peptide domain (NCRPD) . The Orpinomyces xylanase contained one catalytic domain and thus differed from the Neocallimastix xylanase, which had two similar catalytic domains (H . J . Gilbert, G . P . Hazlewood, J . I . Lauie, C . G . Orpin, and G . P . Xue, Mol . Microbiol . 6:2065-2072, 1992) . Two peptides corresponding to the catalytic domain and the NCRPD of XynA were synthesized, and antibodies against them were raised and affinity column purified . The antibodies against the catalytic domain peptide reacted specifically with the xylanases of Orpinomyces and Neocallimastix, while the antibodies against the NCRPD reacted with many (at least eight) extracellular proteins of Orpinomyces and Neocallimastix, suggesting that the NCRPD is present in a number of polypeptides. Appl Environ Microbiol, 1997 Feb, 63(2), 488 - 97 Glucoamylase gene fusions alleviate limitations for protein production in Aspergillus awamori at the transcriptional and (post) translational levels; Gouka RJ et al.; In this study we have analyzed the effects of a glucoamylase gene fusion on the mRNA levels and protein levels for the human interleukin-6 gene (hil6) and the guar alpha-galactosidase gene (aglA) . Previously it was shown that production of nonfused alpha-galactosidase and hIL-6 in Aspergillus awamori was limited at transcriptional and (post)translational levels, respectively (R . J . Gouka, P . J . Punt, J . G . M . Hessing, and C . A . M . J . J . van den Hondel, Appl . Environ . Microbiol . 62:1951-1957, 1996) . Vectors were constructed which contained either the hil6 or aglA gene fused to the Aspergillus niger glucoamylase gene (glaA) under control of the efficient 1,4-beta-endoxylanase A promoter and transcription terminator . For comparison, the vectors were integrated in a single copy at the pyrG locus of A . awamori . A glaA fusion to the 5' end of the hil6 gene resulted in a large increase in hIL-6 yield, whereas with a glaA fusion to the 3' end of the hil6 gene, almost no protein was produced . Nevertheless, the steady-state mRNA levels of both fusions were very similar and not clearly increased compared to those of a strain expressing nonfused hIL-6 . Fusions of glaA to the 5' end of the wild-type guar aglA gene resulted in truncated mRNA lacking almost 900 bases (> 80%) of the aglA sequence . When the coding sequence of the wild-type aglA gene was replaced by a synthetic aglA gene with optimized Saccharomyces cerevisiae codon usage, full-length mRNA was obtained . Compared to a nonfused synthetic aglA gene, a glaA fusion with the synthetic aglA gene resulted in a 25-fold increase in the mRNA level and, as a consequence, a similar increase in the alpha-galactosidase protein level . The truncated transcripts derived from the wild-type aglA gene were further analyzed by nuclear run-on transcription assays . These experiments indicated that transcription elongation in the nucleus proceeded at least 400 bases downstream of the site where the truncation was determined, indicating that transcription elongation or premature termination was not the reason for the generation of truncated mRNAs . As the truncated mRNA also contained a poly(A) tail, truncation most likely occurs by incorrect processing of the aglA mRNA in the nucleus. J Bacteriol, 1997 Feb, 179(4), 1246 - 52 Further characterization and in situ localization of chain-like aggregates of the gliding bacteria Myxococcus fulvus and Myxococcus xanthus; Freese A et al.; For the first time, chain-like aggregates, called "strands," have been enriched from crude cell wall preparations of liquid-grown vegetative cells of two strains of Myxococcus xanthus . These strands are highly isomorphic to macromolecular structures, previously described for Myxococcus fulvus (Lunsdorf and Reichenbach, J . Gen . Microbiol . 135:1633-1641, 1989) . The strands are morphologically composed of ring elements, consisting of six or more peripheral protein masses and possibly three small central masses . The ring elements are linked by two parallel strings of filamentous proteins, called elongated elements, which keep the ring elements at a constant distance . The overall dimensions of the ring elements are 16.6 +/- 1.0 nm (n = 55) for M . xanthus Mx x48 and 16.4 +/- 1.5 nm (n = 37) for M . xanthus DK 1622 . The distance between the ring elements, as a measure of the length of the elongated elements, is 16.6 +/- 1.1 nm (n = 59) for strain Mx x48 and 15.5 +/- 0.6 nm (n = 41) for strain DK 1622 . Characteristically, the strands and oligomeric forms thereof show a strict association with the outer membrane . In situ studies of freeze-fractured cells of M . fulvus showed ring elements, isomorphic to those described for M . xanthus, within the periplasm; they appeared in parallel rows just below the outer membrane but not in direct contact with the cytoplasmic membrane . A three-dimensional model summarizes the morphological data . It is hypothesized that the chain-like strands, as building blocks of a more complex belt-like continuum, represent the peripheral part of the gliding machinery, which transforms membrane potential energy into mechanical work. J Bacteriol, 1997 Feb, 179(3), 919 - 27 Characterization of the p-toluenesulfonate operon tsaMBCD and tsaR in Comamonas testosteroni T-2; Junker F et al.; Comamonas testosteroni T-2 uses a standard, if seldom examined, attack on an aromatic compound and oxygenates the side chain of p-toluenesulfonate (TS) (or p-toluenecarboxylate) to p-sulfobenzoate (or terephthalate) prior to complete oxidation . The expression of the first three catabolic enzymes in the pathway, the TS methyl-monooxygenase system (comprising reductase B and oxygenase M; TsaMB), p-sulfobenzyl alcohol dehydrogenase (TsaC), and p-sulfobenzaldehyde dehydrogenase (TsaD), is coregulated as regulatory unit R1 (H . R . Schlafli Oppenberg, G . Chen, T . Leisinger, and A . M . Cook, Microbiology {Reading} 141:1891-1899, 1995) . The components of the oxygenase system were repurified, and the N-terminal amino acid sequences were confirmed and extended . An internal sequence of TsaM was obtained, and the identity of the {2Fe-2S} Rieske center was confirmed by electron paramagnetic resonance spectroscopy . We purified both dehydrogenases (TsaC and TsaD) and determined their molecular weights and N-terminal amino acid sequences . Oligonucleotides derived from the partial sequences of TsaM were used to identify cloned DNA from strain T-2, and about 6 kb of contiguous cloned DNA was sequenced . Regulatory unit R1 was presumed to represent a four-gene operon (tsaMBCD) which was regulated by the LysR-type regulator, TsaR, encoded by a deduced one-gene transcriptional unit . The genes for the inducible TS transport system were not at this locus . The oxygenase system was confirmed to be a class IA mononuclear iron oxygenase, and class IA can now be seen to have two evolutionary groups, the monooxygenases and the dioxygenases, though the divergence is limited to the oxygenase components . The alcohol dehydrogenase TsaC was confirmed to belong to the short-chain, zinc-independent dehydrogenases, and the aldehyde dehydrogenase TsaD was found to resemble several other aldehyde dehydrogenases . The operon and its putative regulator are compared with units of the TOL plasmid. J Clin Microbiol, 1997 Feb, 35(2), 379 - 82 Gordona terrae central nervous system infection in an immunocompetent patient; Drancourt M et al.; The bacterial genus Gordona includes seven species of mycolic acid-containing actinomycetes which are cultured from environmental sources and rarely from clinical samples . They have been implicated in primary pulmonary diseases, nosocomial wound infections, and central nervous system infections in two immunocompromised patients . We isolated Gordona terrae from the cerebrospinal fluid sample of an immunocompetent patient with meningitis and multiple brain abscesses and detected Gordona terrae DNA in the abscesses . The outcome was good at the 4-year follow-up, after prolonged treatment with trimethoprim-sulfamethoxazole . Phenotypic identification of this isolate was confirmed by analysis of the 16S rRNA gene sequence, which shared 100% homology with that of G . terrae reference strains . Physicians and clinical microbiologists must be aware of the occurrence of Gordona species infection not only among immunocompromised patients but among all patients . Accurate identification of Gordona species may be accomplished by molecular techniques. Gene, 1997 Jan 31, 185(1), 127 - 32 A putative ABC-transport operon of Mycobacterium smegmatis; Barsom EK et al.; We have recently described the mpr gene of Mycobacterium smegmatis whose product confers resistance to mycobacteriophages L5 and D29 when overproduced (Barsom and Hatfull (1996) Mol . Microbiol . 21, 159-170) . We have determined the nt sequence of approximately 3.5 kb immediately adjacent to mpr which appears to encode components of an ATP-binding cassette (ABC) transport system . Four closely-spaced open reading frames (ORF) were identified although two of these may cooperate to produce an integral membrane component of the transport system via a programmed translational frameshift . Another putative protein is also predicted to be an integral membrane protein, while the third is an ABC-transporter protein . We propose that these three putative proteins form a mycobacterial membrane-bound complex involved in protein-dependent transport . This is the first ABC-transport system to be described in mycobacteria. Gene, 1997 Jan 31, 185(1), 27 - 33 Extending filamentous phage host range by the grafting of a heterologous receptor binding domain; Marzari R et al.; fd and IKe are two similar filamentous phage which infect their hosts by means of pili found on the host membrane: fd infects bacteria bearing F pili, whereas IKe infects bacteria bearing N or I pili . Infection is mediated by the gene 3 protein (g3p), which of the nine proteins found in both phage is the most diverse . Previous attempts to incorporate g3p from one phage into the other by complementation have been unsuccessful {Bross et al . (1988) J . Gen . Microbiol . 134, 461-471} . Here we have grafted different parts of IKe g3p to the end of fd g3p and so augmented the host range of fd phage . We show that phage bearing such chimeric g3p are able to infect bacteria bearing both N and F pili providing they contain at least the receptor domain of IKe g3p, the infection of N bearing bacteria occurring at a level 70,000 times greater than background . This level of infection can be increased tenfold by including the glycine-rich domain as well . Addition of the penetration domain does not improve the level of infection above that of the receptor domain alone, indicating that the fd penetration domain is functional in the infection of bacteria bearing either N or F pili . Similarly derived fd phagemid also show increased infection of bacteria bearing N pili, albeit at much lower levels, suggesting that efficient infection requires more than one functional g3p on the surface of the phage. J Exp Biol, 1997, 200(Pt 15), 2137 - 43 Honeydew sugars and osmoregulation in the pea aphid Acyrthosiphon pisum Wilkinson T, Ashford D, Pritchard J, Douglas A. Pea aphids, Acyrthosiphon pisum, containing their symbiotic bacteria (untreated aphids) and experimentally deprived of their bacteria by treatment with the antibiotic rifampicin (antibiotic-treated aphids) were reared on the plant Vicia faba . The sugars in the honeydew produced by untreated aphids comprised predominantly the monosaccharides glucose and fructose, while the honeydew of antibiotic-treated aphids contained considerable amounts of oligosaccharides of up to 16 hexose units . The honeydew and haemolymph of the aphids were iso-osmotic, and their osmotic pressure was significantly lower in untreated aphids (0.91&shy;0.95 MPa) than in antibiotic-treated aphids (1.01&shy;1.05 MPa) (P<0.05) . For insects reared on chemically defined diets containing 0.15&shy;1.0 mol l-1 sucrose (osmotic pressure 1.1&shy;4.0 MPa), the osmotic pressure of the aphid haemolymph did not vary with dietary osmotic pressure, but was regulated to approximately 1.0 MPa in untreated and 1.3 MPa in antibiotic-treated aphids . The sugars in the aphid honeydew varied with dietary sucrose concentration; with monosaccharides dominant at low concentrations and oligosaccharides dominant at high concentrations of dietary sucrose . The lowest dietary sucrose concentration at which honeydew oligosaccharides were detected was 0.2 mol l-1 for the antibiotic-treated aphids and 0.3 mol l-1 for untreated aphids . These data indicate that the aphid, and not its associated microbiota, mediates the synthesis of oligosaccharides when the osmotic pressure of the ingesta is high. Vestn Ross Akad Med Nauk, 1997, (5), 3 - 12 {Nikolaĭ Nikalaevich Sirotinin and his school}; Agadzhanian NA et al.; In a brief review of 50-year scientific activity of professor N . N . Sirotinin and his students the authors emphasize that this broad-minded scientist contributed to development of such disciplines as microbiology, pathophysiology, high-altitude, aerospace medicine . However, his main goal was evolution of reactivity and resistance, approaches to perfection of human health and performance . Much attention was paid to effects of low partial oxygen pressure on human and animal body, to hypoxic states of different origin . Methods of hypoxytherapy and hypoxic training are widely used in Russia and abroad . The contribution of academician N . N . Sirotinin to modern pathophysiology, high-altitude and aerospace physiology, internal and sport medicine is highly appreciated in Russia. Am J Nephrol, 1997, 17(3-4), 347 - 58 The introduction of renal biopsy into nephrology from 1901 to 1961: a paradigm of the forming of nephrology by technology; Cameron JS et al.; 'Biopsy' (Besnier 1895) became useful towards the end of the 19th century with the development of good histology and microbiology . Needle biopsy of the liver, although first performed in 1895, did not become current until 50 years later . Surgical biopsy of the kidney at incidental operations, particularly the then fashionable renal decapsulation, was performed from 1900 to 1930 . Percutaneous needle renal biopsy was introduced after first, the successful liver biopsy and second, demonstration of the value of aspiration needle biopsy in tumours of the kidney . In addition, a number of physicians obtained renal tissue by accident and without problems during intended biopsies of the liver . Nils Alwall of Sweden performed the first systematic aspiration needle biopsies of the kidney in 1944, but did not publish his results because of an early death which led him to abandon the technique . However, when Iversen and Brun in Copenhagen described their results in 1951, a number of physicians around the world immediately began to attempt renal biopsy, using cutting as well as aspiration techniques . Success was inconsistent and operator dependent: the refinements of technique and needles introduced by the group in Chicago led by Robert Kark, plus their advocacy of the technique and their training of many physicians in its performance rapidly led to widespread acceptance . New techniques of immunofluorescence and electron microscopy arrived at the same time so that the technique could be fully exploited . The performance and interpretation of renal biopsies became, along with classical whole-organ and nephron physiology and the introduction of dialysis and transplantation, powerful agents determining the emergence of Nephrology as a specialty around 1960. Pathologe, 1997 Jan, 18(1), 27 - 36 {Mastitis . Classification, histopathology and clinical aspects}; Bassler R; The frequency of biopsies with different types of mastitis among all patients with benign breast diseases in twenty years was 3% . About 2/3 of the patients were younger than 50 years, the average age was 46.7 years . Comparing the frequency of the different types of mastitis the puerperal mastitis is very rare in contrast to the increasing non-puerperal and granulomatous inflammatory breast lesions . The diagnosis "granulomatous mastitis" is one of exclusion . Both non-infectious and infectious causes must be considered . The origin of the granulomatous mastitis often is unknown, but it is supposed to be an autoimmune localized response due to the retained and extravasated fat and protein rich secretions in the ducts in cases of hyperprolactinemia (drug induced or by microprolactinomas) or hormonal imbalances characterized by epitheloid cell granulomas with giant cells microabscesses around lipid drops, but without necroses . An idiopathic type of granulomatous mastitis concerns young women in relationship to parturition with a similar histological pattern predominantly of the lobules . Special types of mastitis are the B-lymphocytic autoimmune mastitis associated with a longstanding insulin-dependent diabetes mellitus type I, the sarcoidosis, panniculitis and the rare but very different infectious diseases with breast involvement . Particularly in cases without features of secretory activity a carefully bacterioscopic and microbiologic work-up of fresh material from the surgical specimens is necessary for a final diagnostic report. Springer Semin Immunopathol, 1997, 18(4), 449 - 61 The intestinal epithelial cell: immunological aspects; Christ AD et al.; IECs likely play an important role in immunological defense mechanism . Apart from being a passive barrier against luminal bacteria, IECs secrete protective and microbiocidal products such as ITF, complement components and cryptdins into the lumen . Moreover, IECs produce secretory component that is essential for the transport of IgA from the lamina propria into the lumen . IECs also have regulatory functions . They express adhesion molecules important in the homing of T cells and other leukocytes, and likely modulate T cell functions in a paracrine way . Furthermore, IECs secrete cytokines, either constitutively or after bacterial challenge, and they express cytokine receptors . Lastly, IECs may play an important role as non-professional antigen-presenting cells by expressing classical MHC class I and class II and nonclassical MHC class I molecules on the cell surface . This aspect is particularly intriguing in that IECs also express a FcR that may have a function in luminal antigen sampling. Jt Comm J Qual Improv, 1997 Jan, 23(1), 23 - 31 Local heroes beat national champions: quality improvement in Finnish health care; Brommels M et al.; FINNISH HEALTH CARE: Although health care services in Finland are organized uniformly throughout the country, they, along with the application of total quality management (TQM)/continuous quality improvement (CQI), are the responsibility of municipalities . CASE 1: At the Helsinki City Health Organization, top management launched an organizationwide quality improvement (QI) initiative in 1993, entailing 150 CQI projects . Yet top and middle managers were not sufficiently dedicated to the initiative to provide adequate support to many of the quality projects . Only "islands of activity" were spotted . CASE 2: A physician in the pediatrics department of a hospital helped initiate CQI projects--for improvements in administration of x-rays for patients with antebrachium fractures, transport of samples to the microbiology laboratory, and admissions of patients with acute infectious disease . Successes led senior management in the hospital federation to issue a quality policy based on CQI management, showing the power of the good example . CASE 3: The first launch of CQI at the Helsinki University Central hospital was part of a management development project that did not have the full support of senior management and that, consequently, failed . A second initiative undertaken a year later, the quality council, was more successful . CASE 4: At a local center for social services and primary care, quality projects have strengthened the organization's team and network structures across two professional cultures . DISCUSSION: The four cases provide insight into the diffusion of TQM/QI and implementation strategies on the local level. Diagn Cytopathol, 1997 Jan, 16(1), 26 - 30 Aspergillus in cytology specimens: a review of 45 specimens from 36 patients; Fischler DF et al.; OBJECTIVE: To assess the clinical significance associated with the identification of fungal elements consistent with Aspergillus in cytology specimens . MATERIALS AND METHODS: For all cytology specimens with reported fungal elements consistent with Aspergillus, reported over a 9 yr and 8 mo period at The Cleveland Clinic Foundation, the patient's medical charts were reviewed with particular attention to underlying disease, presentation, treatment, and clinical course . Cytology results were compared with available microbiologic cultures and tissue specimens in all of the patients . RESULTS: Forty-five cytology specimens with Aspergillus fungal forms, from 36 patients, were identified . Twenty-six patients had concurrent specimens sent for culture in whom II grew Aspergillus species (10 Aspergillus fumigatus), eight grew organisms other than Aspergillus, and seven were no growth . A total of 16 patients (44%) were treated with antifungal treatment (Amphotericin B) . Treatment with Amphotericin B was significantly associated with a concurrent growth of Aspergillus species (9/11 patients with Aspergillus culture positive vs . 7/25 patients without a positive culture for Aspergillus, P value = 0.004 (ODDS ratio = 11, 95% confidence interval:#1.6-104, 2-tailed Fisher exact test.) CONCLUSIONS: The presence of fungal forms consistent with Aspergillus in cytology specimens is neither specific nor sensitive for significant infection due to Aspergillus . Treatment with Amphotericin B is more likely to be instituted when a concurrent clinical specimen grows Aspergillus species in culture. Ophthalmic Surg Lasers, 1997 Jan, 28(1), 43 - 9 Impact of cultures on management decisions following surgical repair of penetrating ocular trauma; Rubsamen PE et al.; BACKGROUND AND OBJECTIVE: The purpose of this study was to evaluate the utility of routine bacterial and fungal cultures in the diagnosis of endophthalmitis and in the subsequent management of patients following penetrating ocular trauma . PATIENTS AND METHODS: The medical records of 70 consecutive patients with penetrating ocular trauma for whom intraoperative bacterial and fungal cultures had been obtained from the wound, aqueous, vitreous, and/or intraocular foreign body (IOFB) were retrospectively reviewed . The incidences of infection among eyes with and without a clinical diagnosis of infection were compared . A determination as to change in clinical management (change of antibiotic, length of treatment) was made . RESULTS: Twenty of 70 patients (29%) had positive cultures of the wound, aqueous, vitreous, and/or IOFB . Nine (13%) of the 70 patients were diagnosed as having endophthalmitis, based on clinical findings at presentation and during the subsequent clinical course . Seven (78%) of these 9 patients with a clinical diagnosis of endophthalmitis had positive cultures . The remaining 61 eyes showed no evidence of clinically apparent infections, despite positive cultures from 13 eyes (21%) . Microbiologic data derived from the culture results influenced the clinical management of all 7 patients with endophthalmitis . Culture results (positive or negative) did not alter clinical decisions in eyes without clinical evidence of infection . CONCLUSION: Bacterial or fungal cultures obtained from the eye in the setting of penetrating trauma often had growth of organisms without clinical signs of infection . Positive culture results directly influenced management decisions in cases with clinically evident endophthalmitis . However, routine intra-operative bacterial cultures did not help to identify patients in whom endophthalmitis would develop, nor did they assist in directing management decisions in eyes without clinical suspicion of infection. J Clin Lab Anal, 1997, 11(1), 2 - 9 In situ hybridization: methods and applications; Jin L et al.; In situ hybridization (ISH) combines molecular biological techniques with histological and cytological analysis of gene expression . RNA and DNA can be readily localized in specific cells with this method . ISH has been useful as a research tool, and recent studies have used this technique in the diagnostic pathology laboratory and in microbiology for the tissue localization in infectious agents . Other recent developments in the applications of ISH involve in situ polymerase chain reaction (PCR) and in situ reverse transcription (RT)-PCR, which can be used to detect very low levels of nucleic acids in tissues by taking advantage of the powerful amplification capacity of PCR . In situ PCR will contribute significantly to progress in this field because of the marked increase in the sensitivity of this method. Clin Diagn Lab Immunol, 1997 Jan, 4(1), 60 - 3 PCR-restriction fragment length polymorphism analysis of the ospC gene for detection of mixed culture and for epidemiological typing of Borrelia burgdorferi sensu stricto; Masuzawa T et al.; Restriction fragment length polymorphism (RFLP) analysis of the outer surface protein C (ospC) gene amplicon was used for rapid screening for genetic variability within Borrelia burgdorferi sensu stricto species and for detection of multiple borreliae in culture . Primers for the ospC gene amplified a fragment of about 600 bp from Borrelia cultures . After cleavage of the amplified products by MboI and DraI, eight different RFLP types were found among 13 B . burgdorferi sensu stricto strains from various sources and geographical areas, and three RFLP types were found among 10 representative isolates from skin biopsy specimens taken from patients residing on the eastern end of Long Island, New York (B . W . Berger, R . C . Johnson, C . Kodner, and L . Coleman, J . Clin . Microbiol . 30:359-361, 1992) . These results suggested that the DNA organization of B . burgdorferi sensu stricto is heterogeneous not only globally but also within a localized geographical area and that the ospC-based typing approach could differentiate the B . burgdorferi sensu stricto . From the results obtained using mixed cultures of two different RFLP types of B . burgdorferi sensu stricto, contamination of at least 0.5% of different types of Borrelia cells in culture could be detected . This method could detect a multiple-B . burgdorferi sensu stricto infection in the bladders of mice experimentally infected with two different RFLP type strains . The present study showed that RFLP analysis of ospC-PCR products is a reliable method for epidemiological typing of B . burgdorferi sensu stricto and could be used for rapid detection of mixed Borrelia culture and multiple B . burgdorferi sensu stricto infections in animals, ticks, and patients. EXS, 1997, 80, 267 - 83 Microbiosensors using electrodes made in Si-technology; Hintsche R et al.; The combination of electrochemical transducers made in silicon technology with chemical and biochemical components has been used to manufacture miniaturized sensor structures . Three different types of sensors have been developed and optimized for practical use; (i) an ion-selective sensor, (ii) a glucose enzyme sensor, (iii) a redox-amplifying sensor for immunosensing . The immunodetection based on the redox recycling of mediator molecules is shown for low and high molecular weight analytes . The sensors have been integrated with miniaturized fluidic components and combined with sensor-related electronics and a common microcontroller. Am J Respir Crit Care Med, 1997 Jan, 155(1), 371 - 3 Pleural effusion due to herpes simplex type II infection in an immunocompromised host; Trudo FJ et al.; Pleural effusions caused by herpes simplex viruses are rare . We report a case of a young woman with acute lymphocytic leukemia (ALL) and prolonged neutropenia who developed pleural space infection with herpes simplex type II virus (HSV II), as confirmed by cytologic and microbiologic studies . We believe that this is the first report of a pleural effusion caused by HSV II, and suggest that this virus now be considered in the differential diagnosis of an unexplained exudative pleural effusion, especially in an immunocompromised host. Int J Syst Bacteriol, 1997 Jan, 47(1), 103 - 10 Investigations of the taxonomy and systematics of Fusobacterium nucleatum using allozyme electrophoresis; Morris ML et al.; Fusobacterium nucleatum forms part of the resident microbiota in both oral and extraoral sites in humans and animals . It is also involved in infections in such sites . Despite the genetic heterogeneity within the species, it has been divided into five subspecies, the validities of which have been questioned . In the present study, 44 F . nucleatum isolates were examined at 21 enzyme loci by using the allozyme electrophoretic technique to establish an accurate genetic framework for taxonomic purposes . Three distinct genetic clusters were identified; one cluster consisted exclusively of extraoral isolates, another cluster consisted predominantly of human oral isolates, and the third cluster consisted of a single human oral isolate . Our results highlight the urgent need for extensive biochemical, immunological, and epidemiological studies to accurately define the systematics of the genus fusobacterium based on the framework derived in this study by using 21 independent genetic characteristics. J Bacteriol, 1997 Jan, 179(2), 530 - 7 Function of the C-terminal domain of the alpha subunit of Escherichia coli RNA polymerase in basal expression and integration host factor-mediated activation of the early promoter of bacteriophage Mu; van Ulsen P et al.; Integration host factor (IHF) can activate transcription from the early promoter (Pe) of bacteriophage Mu both directly and indirectly . Indirect activation occurs through alleviation of H-NS-mediated repression of the Pe promoter (P . Van Ulsen, M . Hillebrand, L . Zulianello, P . Van de Putte, and N . Goosen, Mol . Microbiol . 21:567-578, 1996) . The direct activation involves the C-terminal domain of the alpha subunit (alphaCTD) of RNA polymerase . We investigated which residues in the alphaCTD are important for IHF-mediated activation of the Pe promoter . Initial in vivo screening, using a set of substitution mutants derived from an alanine scan (T . Gaal, W . Ross, E . E . Blatter, T . Tang, X . Jia, V . V . Krishnan, N . Assa-Munt, R . Ebright, and R . L . Gourse, Genes Dev . 10:16-26, 1996; H . Tang, K . Severinov, A . Goldfarb, D . Fenyo, B . Chait, and R . H . Ebright, Genes Dev . 8:3058-3067, 1994), indicated that the residues, which are required for transcription activation by the UP element of the rrnB P1 promoter (T . Gaal, W . Ross, E . E . Blatter, T . Tang, X . Jia, V . V . Krishnan, N . Assa-Munt, R . Ebright, and R . L . Gourse, Genes Dev . 10:16-26, 1996), are also important for Pe expression in the presence of IHF . Two of the RNA polymerase mutants, alphaR265A and alphaG296A, that affected Pe expression most in vivo were subsequently tested in in vitro transcription experiments . Mutant RNA polymerase with alphaR265A showed no IHF-mediated activation and a severely reduced basal level of transcription from the Pe promoter . Mutant RNA polymerase with alphaG296A resulted in a slightly reduced transcription from the Pe promoter in the absence of IHF but could still be activated by IHF . These results indicate that interaction of the alphaCTD with DNA is involved not only in the IHF-mediated activation of Pe transcription but also in maintaining the basal level of transcription from this promoter . Mutational analysis of the upstream region of the Pe promoter identified a sequence, positioned from -39 to -51 with respect to the transcription start site, that is important for basal Pe expression, presumably through binding of the alphaCTD . The role of the alphaCTD in IHF-mediated stimulation of transcription from the Pe promoter is discussed. J Bacteriol, 1997 Jan, 179(1), 163 - 9 Gene transcription and chromosome replication in Escherichia coli; Zhou P et al.; Transcript levels of several Escherichia coli genes involved in chromosome replication and cell division were measured in dnaC2(Ts) mutants synchronized for chromosome replication by temperature shifts . Levels of transcripts from four of the genes, dam, nrdA, mukB, and seqA, were reduced at a certain stage during chromosome replication . The magnitudes of the decreases were similar to those reported previously ftsQ and ftsZ (P . Zhou and C . E . Helmstetter, J . Bacteriol . 176:6100-6106, 1994) but considerably less than those seen with dnaA, gidA, and mioC (P . W . Theisen, J . E . Grimwade, A . C . Leonard, J . A . Bogan, and C . E . Helmstetter, Mol . Microbiol . 10:575-584, 1993) . The decreases in transcripts appeared to correlate with the estimated time at which the genes replicated . This same conclusion was reached in studies with synchronous cultures obtained with the baby machine in those instances in which periodicities in transcript levels were clearly evident . The transcriptional levels for two genes, minE and tus, did not fluctuate significantly, whereas the transcripts for one gene, iciA, appeared to increase transiently . The results support the idea that cell cycle timing in E . coli is not governed by timed bursts of gene expression, since the overall findings summarized in this report are generally consistent with cell cycle-dependent transient inhibitions of transcription rather than stimulations. Infect Immun, 1997 Jan, 65(1), 331 - 4 Protease susceptibility and toxicity of heat-labile enterotoxins with a mutation in the active site or in the protease-sensitive loop; Giannelli V et al.; To generate nontoxic derivatives of Escherichia coli heat-labile enterotoxin (LT), site-directed mutagenesis has been used to change either the amino acid residues located in the catalytic site (M . Pizza, M . Domenighini, W . Hol, V . Giannelli, M . R . Fontana, M . M . Giuliani, C . Magagnoli, S . Peppoloni, R . Manetti, and R . Rappuoli, Mol . Microbiol . 14:51-60, 1994) or those located in the proteolytically sensitive loop that joins the A1 and A2 moieties of the A subunit (C . C . R . Grant, R . J . Messer, and W . J . Cieplack, Infect . Immun . 62:4270-4278, 1994; B . L . Dickinson and J . D . Clements, Infect . Immun . 63:1617-1623, 1995) . In this work, we compared the in vitro and in vivo toxic properties and the resistance to protease digestion of the prototype molecules obtained by both approaches (LT-K63 and LT-R192G, respectively) . As expected, LT-K63 was normally processed by proteases, while LT-R192G showed increased resistance to trypsin in vitro and was digested by trypsin only under denaturing conditions (3.5 M urea) or by intestinal proteases . No toxicity was detected with the LT-K63 mutant, even when 40 micrograms and 1 mg were used in the in vitro and in vivo assays, respectively . In marked contrast, LT-R192G showed only a modest (10-fold) reduction in toxicity in Y1 cells with a delay in the appearance of the toxic activity and had toxicity comparable to that of wild-type LT in the rabbit ileal loop assay . We conclude that mutagenesis of the active site generates molecules that are fully devoid of toxicity, while mutagenesis of the A1-A2 loop generates molecules that are resistant to trypsin in vitro but still susceptible to proteolytic activation by proteases other than trypsin, and therefore they may still be toxic in tissue culture and in vivo. J Clin Microbiol, 1997 Jan, 35(1), 217 - 22 Capture-S, a nontreponemal solid-phase erythrocyte adherence assay for serological detection of syphilis; Stone DL et al.; A solid-phase erythrocyte adherence assay has been developed for the serological detection of reagin antibodies in syphilis . Capture-S (Immucor, Inc., Norcross, Ga.) is a nontreponemal, qualitative screening test for the detection of immunoglobulin G (IgG) and IgM antilipid antibodies in serum or plasma samples from blood donors . The Capture-S assay utilizes a modified Venereal Disease Research Laboratory antigen bound to microtitration wells and anti-IgG- plus anti-IgM-coated indicator erythrocytes as the detection system . The Capture-S assay was evaluated at six separate sites on 10,942 specimens . For patient samples of clinically diagnosed syphilis categories (n = 366), the Capture-S assay yielded a sensitivity of 80.7% versus 80.3% for the rapid plasma reagin (RPR) card test (Becton Dickinson Microbiology Systems, Cockeysville, Md.) . In comparative experiments on patient and donor samples (n = 10,222), the Capture-S assay demonstrated a sensitivity of 94% compared to 91.2% for the RPR card test . The Capture-S and RPR card tests produced essentially equivalent specificities of 99.2% and 99.3%, respectively, for this sample population . For five test sites, the Capture-S and RPR card test demonstrated a 98.3% agreement (10,085 of 10,264) of test results . These evaluations indicate that the Capture-S compares favorably to the RPR card test in assay sensitivity and specificity, with the added benefits of ease of use, accommodation of high-volume testing, and potential for automation. J Clin Microbiol, 1997 Jan, 35(1), 117 - 20 Detection of Bordetella pertussis in clinical specimens by PCR and a microtiter plate-based DNA hybridization assay; Nelson S et al.; In order to improve detection of Bordetella pertussis in nasopharyngeal aspirates (NPAs) in our laboratory, a PCR-based assay was optimized, and a study was designed (i) to compare results obtained by PCR to those obtained by culture and (ii) to evaluate a novel microtiter plate-based DNA hybridization assay (PCR-plate) by comparing it to agarose gel electrophoresis (PCR-gel) for detection of the PCR product . DNA for the PCR was extracted with a guanidine thiocyanate buffer and used in a PCR mixture containing primers directed against a reiterated gene sequence in B . pertussis (Q . He, J . Mertsola, H . Soini, M . Skurnik, O . Ruuskanen, and M . K . Viljanen, J . Clin, Microbiol . 31:642-645, 1993) . Of 96 NPAs submitted from a targeted study group, 23 were positive by culture, 27 were positive by PCR-gel, and 31 were positive by PCR-plate . All culture-positive specimens were also positive by PCR . Of nine patients with culture-negative-PCR-positive results, six had discharge diagnoses of pertussis . Thus, PCR with plate-based product detection is a sensitive method for the laboratory detection of B . pertussis in NPAs . Additional advantages of the plate assay include rapidity, objectivity in reading results, specificity, and the capability of being adapted to a high-volume, automated system. J Clin Microbiol, 1997 Jan, 35(1), 25 - 32 Genotypic characterization of five subspecies of Mycobacterium kansasii; Picardeau M et al.; Different molecular typing methods including restriction fragment length polymorphism (RFLP) analysis with the major polymorphic tandem repeat (MPTR) probe and the IS1652 probe, pulsed-field gel electrophoresis (PFGE), amplified fragment length polymorphism (AFLP) analysis, and PCR restriction analysis of the hsp-65 gene (PRA) were applied to clinical and water isolates of Mycobacterium kansasii . RFLP with the MPTR probe, PRA, PFGE, and AFLP analysis revealed five homogeneous clusters which appeared to be subspecies . RFLP with the MPTR probe and PRA gave patterns specific for each cluster, whereas PFGE and AFLP analysis gave polymorphic patterns . IS1652 was present in two of the five clusters and provided polymorphic patterns for one cluster only . The two IS1652-positive clusters were Accuprobe negative (Accuprobe test; Gen-Probe Inc.), and only two other clusters were Accuprobe positive . A PCR test based on the detection of a species-specific fragment (M . Yang, B.C . Ross, and B . Dwyer, J . Clin . Microbiol . 31:2769-2772, 1993) was positive for all M . kansasii strains . This PCR test is an accurate, rapid, and specific M . kansasii identification test . No subspecies was particularly more virulent, because all clusters contained clinical strains, from AIDS patients and non-AIDS patients, and environmental strains. Biochemistry, 1996 Dec 17, 35(50), 16264 - 9 Kinetic mechanism of the Escherichia coli UDPMurNAc-tripeptide D-alanyl-D-alanine-adding enzyme: use of a glutathione S-transferase fusion; Anderson MS et al.; The D-alanyl-D-alanine-adding enzyme encoded by the murF gene catalyzes the ATP-dependent formation of UDP-N-acetylmuramyl-L-gamma-D-Glu-meso-diaminopimelyl-D-Ala-D-Ala (UDP-MurNAc-tripeptide) . MurF has been cloned from Escherichia coli and expressed as a glutathione S-transferase (GST) fusion using the tac promoter-based pGEX-KT vector . From induced, broken cell preparations, highly active fusion was recovered and purified in one step by affinity chromatography . The purified fusion protein was strongly inhibited by substrate UDPMurNAc-tripeptide, a response unaltered by changes in assay pH or by cleavage from the fusion partner . However, this effect was suppressed by the addition of 0.5 M NaCl . Initial velocity and dead-end inhibitor studies with the fusion enzyme were most consistent with a sequential ordered kinetic mechanism for the forward reaction in which ATP binds to free enzyme, followed by tripeptide and D-Ala-D-Ala in sequence prior to product release . Reported homologies between the MurF protein and the three preceding steps of cytoplasmic murein biosynthesis, MurC, -D, and -E, {Ikeda et al . (1990) J . Gen . Appl . Microbiol . 36, 179-187}, raise the prospect that all of these enzymes will be found to proceed via this mechanism. JAMA, 1996 Dec 4, 276(21), 1737 - 42 Epidemiologic and microbiologic correlates of Chlamydia trachomatis infection in sexual partnerships; Quinn TC et al.; OBJECTIVES: To determine the frequency of Chlamydia trachomatis genital infection within sexual partnerships using highly sensitive polymerase chain reaction (PCR) amplification and to identify the variables that might modify transmission . DESIGN: Cross-sectional study of sexual partnerships comparing in vitro culture and PCR amplification for C trachomatis . SETTING: Two outpatient sexually transmitted disease clinics . PARTICIPANTS: Four hundred ninety-four people in sexual partnerships attending sexually transmitted disease clinics . MAIN OUTCOME MEASURE: Genital infection with C trachomatis . METHODS: DNA sequencing was performed to examine specific genotypes within and between partnerships . Cross-sectional analysis was performed to determine characteristics associated with concordance or discordance of infection with partnerships . RESULTS: Cultures were positive for C trachomatis in 8.5% of males and 12.9% of females (P=.03) . Using PCR, more infections were identified both in males (14.2%) and in females (15.8%), and the difference in infection rates analyzed by sex was no longer significant . In 20.4% of 494 couples, at least 1 partner had PCR results positive for C trachomatis, with a concordant infection rate of 10.7%, significantly higher than the 5.5% concordant infection rate demonstrable by culture (P<.01) . Male-female and female-male transmission frequencies were equal (68%) . The nucleotide sequences of the major outer membrane protein gene products were identical and unique for each of 15 culture-negative, PCR-positive concordant partnerships . In concordant infections, factors associated with infection in female partners were age less than 20 years, more than 1 partner in the past 6 months, and cervical ectopy greater than 25% . CONCLUSIONS: Using PCR, the frequency of chlamydia transmission by infected males and females was nearly identical . The high rate of concordant infection, high frequency of infection among asymptomatic individuals, and high frequency of transmission regardless of sex underscore the importance of routine screening for chlamydia in both males and females, along with provision of treatment to all sexual partners of chiamydia-infected individuals. Singapore Med J, 1996 Dec, 37(6), 631 - 5 Treatment of complicated parapneumonic effusions and pleural empyema: a four-year prospective study; Chin NK et al.; We studied prospectively the microbiologic findings, management, including the use of intrapleural streptokinase to improve pleural drainage, and outcome of 33 patients with complicated parapneumonic effusions (6) frank empyema (27) treated by our unit over a 4-year period . The mean age was 61 years, with more male (26) than female (7) patients . Seventy-nine percent of the patients had some form of underlying illness, especially chronic lung disease (46%), followed by diabetes mellitus (23%) . Fifteen percent of the pleural collections were loculated . Pleural fluid cultures were positive in 58%, Staph aureus and Kleb pneumoniae being the most prevalent aerobic isolates . The incidence of anaerobic isolates was 32% . Besides empirical antibiotics, all patients had drainage of the pleural collections at diagnosis . Four patients were treated with needle aspiration; 28 (85%) required thoracostomy tube drainage for a mean of 8 days, 5 of these went on to surgical decortication . Thirteen patients received intrapleural streptokinase (SK) to facilitate drainage, with significant increase in the volume drained . The mean duration of stay for the whole group was 22 days . The administration of intrapleural SK did not significantly shorten the duration of hospital stay . There were six deaths (18%), none as a direct result of the empyema . We describe a therapeutic approach to parapneumonic effusions and empyema which tailors the interventional modality to the clinical stage of disease. Microbiologia, 1996 Dec, 12(4), 621 - 30 {Brief history of the Spanish Society of Microbiology . VI . From 1991 to 1995}; Garcia Mendoza C; In this sixth and final chapter of the brief history of the Spanish Society for Microbiology (SEM), the major activities carried out from 1991 to 1995 are described . During that period, the 14th and 15th SEM National Congresses took place in Saragossa (1993) and Madrid (1995), respectively . The 1995 Congress was also the occasion to celebrate the fiftieth anniversary of the SEM . During that period, three new Specialized Groups were created: Molecular Microbiology . Aquatic Microbiology, and Protozoology . All Specialized Groups held their own meetings . Courses on the introduction to research in microbiology, for undergraduate students, continued, as well as the Awards of the SEM for young researchers . During that period Cesar Nombela was elected FEMS President, and Fernando Laborda was elected President of the Biodeterioration Society . The photographs of the Presidents and Secretaries of the SEM since its foundation until 1995 are included. J Med Assoc Thai, 1996 Dec, 79 Suppl 1, S78 - 87 Reliable sex determination of mouse preimplantation embryos by PCR amplification of male-specific genes in single blastomeres; Tangchaisin P et al.; OBJECTIVE: To assess the reliability of sex determination in mouse preimplantation embryos using the two-step polymerase chain reaction method . SETTING: Division of Immunology, Department of Microbiology and Division of Reproductive Medicine, Department of OB/GYN . METHODS: The Sry and Zfy genes, known to be present in the sex-determining region of mouse Y chromosome, were selected for Y-specific target sequences and DXNds 3 locus located on mouse X chromosome was served as the internal control sequence . DNAs extracted from heart blood of male and female mice were used to test the correctness and specificity of the selected primers using the two-step PCR method . The same experimental conditions were then used to amplify the single copy genes in single mouse blastomeres with two pairs of primers for each of the target sequences . The sex-determined embryos were transferred to the uteri of pseudopregnant recipients to test the consistency of the assay system . RESULTS: All male and female blood DNA sample results confirmed the correct sex identification of the origin (100%) . Nineteen of 20 single blastomeres showed the accurate diagnosis when compared with theirs 7/8 embryos . The sex of 36 of 37 mouse pups born from biopsied male and female embryos agreed with the predicted sex . CONCLUSION: The reliable genetic analysis of sex chromosome- specific sequences in single cell is possible by the two-step PCR method and could be applied for diagnosis of defective genes of human preimplantation embryos derived from the in vitro fertilization program. Arch Bronconeumol, 1996 Dec, 32(10), 500 - 4 {Computed tomography and polymerase chain reaction in tuberculosis infection in childhood}; Gomez-Pastrana Duran D et al.; The recognition of children with tuberculous infection without disease is often difficult . Minimal active disease may be present in many cases but unrecognised on chest radiography or by microbiologic methods . We have performed computed tomography in 22 children with tuberculous infection, a normal chest radiograph and negative microbacterial culture . In 16 children we also performed DNA amplification by polymerase chain reaction in gastric aspirates . It was found that 14 of 22 (63%) infected children had enlarged lymph nodes . Adenopathies were more frequent in children less than 8 years-old and in the right paratracheal positions . Polymerase chain reaction was positive in 4 of 8 studied children with abnormal computed tomography and in none of the children with normal computed tomography . The demonstration of unrecognised active disease raises the question of the adequate treatment for the children with tuberculous infection . It is proposed that a two drug regimen would be more appropriate than isoniazid alone in children less than 8 years old. Aust Fam Physician, 1996 Dec, 25(12), 1841 - 5 Tick-borne diseases in Australia; Playford G et al.; Tick bites are a common problem in Australia and an important cause of morbidity in medical and veterinary practice . Complications include local inflammation and infection, paralysis and transmission of various pathogens . Over the past three decades, several new tick-borne diseases have been recognised both in Australia and overseas . The importance of these diseases has also increased, in part due to greater recreational activities occurring in tick infested areas . However, our understanding of the microbiology and epidemiology of many of these diseases is incomplete. Aust Fam Physician, 1996 Dec, 25(12), 1831 - 9 The management of common ocular infections; Ng EW et al.; Bacterial and viral infections involving the anterior segment of the eye and ocular adnexa present commonly in general practice, affecting patients of all ages . Careful assessment of the patient is necessary to distinguish between benign conditions and those with more serious complications . An understanding of the microbiology and pathogenesis of these conditions is helpful in rationalising current therapeutic approaches . This article focuses on the clinical presentation of common infections of the lids, conjunctiva and cornea, with a discussion of relevant therapeutic regimens for each condition. J Formos Med Assoc, 1996 Dec, 95(12), 917 - 22 Clinical manifestations, microbiology and prognosis of 42 patients with necrotizing fasciitis; Hung CC et al.; Forty-two cases of necrotizing fasciitis (NF) surgically confirmed between January 1991 and October 1995 were retrospectively reviewed . This was done in order to describe the underlying diseases, clinical presentations, etiology and outcome of NF and to assess the prognostic value of a simplified severity scoring system . The system scores changes in consciousness status, body temperature, blood pressure and ventilation to determine the likely outcome of NF . Twenty-five men and 17 women with a median age of 51 years (range, 17-87 yr) were included . Diabetes mellitus (57.1%) was the most common underlying disease . The mean duration of symptoms before admission was 8 days (median, 7 d; range, 1-30 d) . The extremities (66.7%) were most commonly involved . Initial clinical presentations within 48 hours of admission included skin erythema and swelling at the affected site (97.6%), pyrexia (61.9%), hypotension (33.3%), altered consciousness (28.6%), bullous lesions (26.2%) and crepitus (9.5%) . The mean number of isolated pathogens was 1.8 (range, 0-6) . Eight patients had mixed aerobic and anaerobic infections . The attributable case fatality rate was 23.8% . Higher severity score (> or = 4 points), hypotension, altered consciousness, respiratory failure requiring ventilator support, elevation of alanine aminotransferase levels > twofold, serum creatinine > 177 mumol/L, thrombocytopenia (< 100 x 10(9)/L), and worsening symptoms and signs within 48 hours of admission were associated with higher fatality rates (p < 0.05). Semin Respir Infect, 1996 Dec, 11(4), 231 - 43 Microbiology of nontuberculosis mycobacteria; Gangadharam PR; Recognition of differences, particularly in color and other growth characteristics, between Mycobacterium tuberculosis and other related organisms has led to the identification of the nontuberculosis mycobacteria (NTM) . The Runyon Classification, based largely on the presence (or absence) of pigment and rate of growth, was a convenient system for classifying these organisms . Subsequent biochemical analysis has facilitated more precise speciation . However, many of these tests are complex and time consuming and beyond the capacity of most laboratories other than reference centers . The need for accurate identification has become clear because of differences in pathogenicity and treatment between various species and even subspecies of NTM . The recent appearance of rapid diagnostic tests and highly specific genetic identification systems have created new opportunities and challenges with respect to the identification of NTM . This article provides a historic context for the microbiologic study of the organisms, describes key features of the most commonly encountered NTM, notes several important new technical innovations in this field, and considers common clinical issues in relation to the microbiology of NTM. J Hosp Infect, 1996 Dec, 34(4), 247 - 65 RAPD typing in microbiology--a technical review; Power EG; Many biochemical and molecular techniques can be used for distinguishing isolates of a given bacterial species . Traditional typing techniques based on phenotypic characteristics such as serotyping are being increasingly challenged by the use of DNA-based methods . The introduction of the polymerase chain reaction (PCR) has led to typing techniques based on DNA amplification . Randomly amplified polymorphic DNA (RAPD) typing (also known as arbitrarily primed-polymerase chain reaction, APPCR) is one such technique which is being used increasingly to type micro-organisms, especially during clinical outbreaks . The application and potential problems and solutions of RAPD typing are discussed and the role of such techniques among established typing methods is addressed. J Bacteriol, 1996 Dec, 178(23), 6930 - 6 Effects of nutrition and growth rate on Lrp levels in Escherichia coli; Landgraf JR et al.; Lrp (leucine-responsive regulatory protein) activates some Escherichia coli operons that function in anabolism and represses others involved in catabolism (for a review, see J . M . Calvo and R . G . Matthews, Microbiol . Rev . 58:466-490, 1994) . This overall pattern suggests that Lrp may help cells adapt to changes in the nutritional environment . Here, we tested the idea that the nutritional richness of the medium determines the amount of Lrp in cells . Lrp was measured directly by Western blotting (immunoblotting) in cells grown in a chemically defined rich medium or in a minimal medium . In addition, transcription from the lrp promoter was assessed with a lacZ reporter gene . The results with these two different measurements were nearly the same, indicating that under the conditions employed, beta-galactosidase measurements can accurately reflect Lrp levels . For cells in a minimal medium, Lrp levels were consistently lowest during the logarithmic phase of growth, but overall, there was not much variation in levels as a function of growth phase (1.3-fold difference between highest and lowest values) . However, for cells in a rich medium, Lrp levels dropped 3- to 4-fold during the lag phase, remained constant during the log phase, and then rose to starting levels upon entry into the stationary phase . When cells in the log phase were compared, Lrp levels were 3- to 4-fold higher in cells growing in a minimal medium than those in a rich medium . The levels of lrp expression were the same or slightly higher in strains containing mutations in rpoS, cya, or crp compared with wild-type strains, suggesting that neither RpoS nor the cyclic AMP (cAMP) receptor protein-cAMP complex is required for expression . On the other hand, lrp expression was severely restricted in cells that could not make ppGpp because of mutations in relA and spoT . The reduced expression of lrp during logarithmic growth in a rich medium may be due to low ppGpp levels under these conditions . The repressive effects of rich medium and the stimulatory effects of ppGpp were also observed with a construct having only a minimal lrp promoter (-57 to +21) . The results of other experiments suggest that Lrp levels vary inversely with the growth rate of cells instead of being determined by some component of the medium. Infect Immun, 1996 Dec, 64(12), 5111 - 6 Surface exposure and species specificity of an immunoreactive domain of a 66-kilodalton outer membrane protein (P66) of the Borrelia spp . that cause Lyme disease; Bunikis J et al.; A chromosomally encoded 66-kDa protein (P66) of Borrelia spp . that cause Lyme disease has previously been shown to be associated with the spirochetal outer membrane . A topological model of P66 predicts a surface-exposed fragment which links the N- and C-terminal intramembranous domains of the protein (J . Bunikis, L . Noppa, and S . Bergstrom, FEMS Microbiol . Lett . 131:139-145, 1995) . In the present study, an immunogenic determinant of P66 was identified by a comparison of the immunoreactivities of different fragments of P66 generated either by proteolytic treatment of intact spirochetes or as recombinant proteins expressed in Escherichia coli . The immune response to P66 during natural infection was found to be directed against the predicted surface domain which comprises amino acids at positions 454 through 491 . A sequence comparison revealed considerable polymorphism of the surface domains of P66 proteins of different Lyme disease-causing Borrelia species . Five sequence patterns of this domain were observed in the B . garinii strains studied . In contrast, sequences of the relevant part of P66 of the B . afzelii and B . burgdorferi sensu stricto isolates studied were identical within the respective species . In immunoblotting, 5 of 17 (29.4%) sera from North American patients with early disseminated or persistent Lyme disease reacted against P66 of B . burgdorferi sensu stricto B31 . These sera, however, failed to recognize P66 of B . afzelii and B . garinii, as well as an analog of P66 in the relapsing fever agent, B . hermsii . In conclusion, the topological model of P66 is supported by the demonstration of an apparent surface localization of an immunoreactive domain of this protein . Furthermore, analogous to the plasmid-encoded borrelial outer surface proteins, the predicted surface-exposed portion of chromosomally encoded P66 appears to be antigenically heterogenous. Indian Pediatr, 1996 Nov, 33(11), 905 - 9 Endoscopic, histologic and microbiologic evaluation of upper abdominal pain with special reference to Helicobacter pylori infection; Kumar M et al.; OBJECTIVE: To study children with significant upper abdominal pain of unidentifiable etiology and evaluate: (a) the relationship of pain to inflammatory esophago-gastro-duodenal lesions and Helicobacter pylori (HP) infection, and (b) the response to specific therapy . DESIGN: Prospective study . SETTING: Pediatric section of a tertiary referral gastroenterology center . SUBJECTS: Thirty three consecutive children with significant upper abdominal pain {mean age 9.9 +/- 2.7, range 4-15 years; 20 males} were subjected to upper gastrointestinal tract endoscopy and antral mucosal biopsies obtained for rapid urease test (RUT), Gram's staining of impression/crush smears and culture for HP and histologic examination . Patients with HP gastritis were treated with triple therapy, colloidal bismuth subcitrate, amoxycillin and metronidazole, for two weeks . At 8 weeks from the initiation of therapy, patients were re-evaluated for symptoms and HP eradication by repeat endoscopy and antral biopsies . Patients with esophagitis, gastritis and duodenitis without HP infection were treated with ranitidine for 6 weeks . All the patients were followed up for 6 months . RESULTS: Histology revealed antral gastritis in 28/33 (85%) patients . HP infection was present in 12/28 (43%) patients with antral gastritis . Symptomatic improvement with triple therapy was observed in 10/12 (83%) patients with HP gastritis and eradication of HP in 5/7 . Improvement on ranitidine therapy was observed in 12/16 (75%) patients with HP negative gastritis . On follow-up, no patient with initial improvement with therapy had relapse of symptoms . CONCLUSION: Symptomatic children with HP related gastritis should be treated with triple therapy and HP negative gastritis with H2-receptor antagonist. Aktuelle Radiol, 1996 Nov, 6(6), 308 - 16 {Infections of the cervical spine}; Heindel W et al.; Diagnosis of infectious disease of the spine in an early stage ist difficult . Conventional X-ray examinations, often used as a basic screening study, will show the characteristic narrowing of the intervertebral disc space and the osteolytic and sclerotic changes in the adjointing vertebra only after two to eight weeks . Magnetic resonance imaging (MRI) has a sensitivity equivalent to bone scintigraphy and, due to its superior delineation of anatomic details, MRI has become the method of choice not only for the assessment of location, extent, and associated lesions in infectious bone disease, but also in the diagnosis of intradural infections including myelitis . MRI examination in the assessment of a successful therapy or osteomyelitis of the spine aims for the detection of regression of bone marrow edema, reappearance of fatty marrow, and less pronounced contrast enhancement . Computed tomography (CT) is mainly used for image-guided biopsy to obtain specimen for microbiologic culture or for the placement of a percutaneous drainage. Enferm Infecc Microbiol Clin, 1996 Nov, 14(9), 538 - 40 {Colonic tuberculosis as a cause of rectal bleeding in 2 patients with HIV infection}; Pintor E et al.; BACKGROUND: Extrapulmonary tuberculosis is a relatively frequent disease in patients with HIV infection . This may be observed coinciding with pulmonary involvement or without diagnostic data of the same . Within extrapulmonary involvement, gastrointestinal involvement is one of the least frequently observed and its diagnosis is very difficult due to the scarce clinical suspicion because of the unspecific symptomatology . METHODS: Segmentary colonic tuberculosis was diagnosed in two patients presenting rectal bleeding . The diagnosis was neither clinically nor endoscopically suspected and only histologic and microbiologic study of the biopsies led to establishment of the diagnosis . Pulmonary involvement was also seen in one of the 2 patients . RESULTS: Medical treatment was initiated following the diagnosis . The evolution was good in one patient, while the other developed a picture of massive rectal bleeding requiring surgical resection following which the patient died due to complications . CONCLUSIONS: Colonic tuberculosis is an infrequent disease in patients with HIV infection . Only a high index of clinical suspicion along with adequate histopathologic and microbiologic study of all the colonic lesion biopsies can establish the real frequency of this disease in this group of patients. Rev Prat, 1996 Nov 1, 46(17), 2089 - 92 {Pulmonary Chlamydia infections in children}; Pautard JC; In the last decades chlamydial infection of the respiratory tract have been described in children . Chlamydia trachomatis, the most common identifiable cause of neonatal infections conjunctivitis is also a recognised cause of pneumonia in newborns and is responsible for about one third of sexually transmitted disease . Chlamydia psittaci is mainly pathogenic for birds and occasionally for human . More recently Chlamydia pneumoniae has been found associated with pulmonary abnormalities, nasopharynx and pulmonary oropharynx infections . Microbiologic diagnosis can be difficult . Several non culture methods are now available such as the direct fluorescent antibody test and enzyme immuno-assays . X ray chest show a reticulated and (or) nodular pattern . Treatment is mainly macrolide based with erythromycin. Acta Cytol, 1996 Nov-Dec, 40(6), 1198 - 204 Histologic processing of thick tissue specimens from cytology slides . A novel technique; Verbeek DH et al.; OBJECTIVE: To describe a novel method by which small tissue fragments found in routine cytology specimens can be processed for histologic slides without destroying or damaging the cytologic specimen . STUDY DESIGN: To develop a method by which small tissue fragments (microbiopsies) are removed from cytologic slides and processed through paraffin or plastic, after which histologic slides can be prepared . RESULTS: In all cases described, the histologic sections from the microbiopsies were of excellent quality . Microscopic examination gave additional important information . Furthermore, immunochemistry could be performed on paraffin-processed microbiopsies . CONCLUSION: This method can be of great value in cytologic specimens in which microbiopsies are present, not only in those cases in which cytologic information is insufficient for diagnosis, but also in cases with conclusive diagnostic information . Routinely applied, this method could make biopsies unnecessary in some cases. Med Hypotheses, 1996 Nov, 47(5), 405 - 7 Immunological aspects of nitric oxide in HIV-1 infection; Torre D et al.; Nitric oxide is produced in large amounts during host defense and immunological reactions and it is likely to have a role in non-specific immunity: nitric oxide exerts microbiostatic and microbicidal activity against a variety of pathogens, including protozoa, fungi, bacteria and some viruses . HIV-1 stimulates nitric oxide production by human macrophages and its production is increased in patients with HIV-1 infection . It is postulated that nitric oxide may play a part in modulating the immune response during HIV-1 infection . Nitric oxide produced by the HIV-1 infected monocytes/macrophages of lymph nodes, may adversely affect the survival of activated immune cells, including B and T lymphocytes and dendritic cells within their vicinity . It is suggested here that production of large amounts of nitric oxide by macrophages may lead to the inactivation of lymphocytes and thus to the induction of a persistent immunosuppression. Sex Transm Dis, 1996 Nov-Dec, 23(6), 498 - 503 Sexual partner networks in the transmission of sexually transmitted diseases . An analysis of gonorrhea cases in Sheffield, UK; Ghani AC et al.; BACKGROUND AND OBJECTIVES: Routine contact tracing data on patients with gonorrhea are used to identify sexual partner networks . These are combined with gonococcal typing data to study patterns of transmission . The role of persons in transmission is discussed . STUDY DESIGN: Contact tracing data on patients with gonorrhea attending the Royal Hallamshire Hospital in Sheffield in 1988 and 1989 are analyzed . Gonococcal strains identified by auxotype/serovar (A/S) class are combined with these data to identify transmission paths . RESULTS: The network contained 1,272 persons, 724 (77%) of whom had gonorrhea during the study period . Four hundred two clusters of linked cases were identified . The largest cluster, containing 35 persons connected over 16 months, is discussed in greater detail to illustrate how these data may help identify patterns of transmission and the role of persons . CONCLUSION: Contact tracing data can be used to identify sexual partner networks and to study transmission patterns . Microbiologic data can aid interpretation . An person's risk of acquiring infection depends on indirect links as well as direct links . To understand patterns of transmission it may be important to distinguish between those involved in transmission and those only acquiring infection . Networks established through gonococcal transmission are also relevant to the transmission of other sexually transmitted diseases. Ophthalmology, 1996 Nov, 103(11), 1854 - 62; discussion 1862-3 Comparison of ciprofloxacin ophthalmic solution 0.3% to fortified tobramycin-cefazolin in treating bacterial corneal ulcers . Ciprofloxacin Bacterial Keratitis Study Group; Hyndiuk RA et al.; PURPOSE: The purpose of the study is to compare the clinical efficacy and safety of ciprofloxacin ophthalmic solution 0.3% (Ciloxan) with a standard therapy regimen (fortified tobramycin, 1.3%-cefazolin, 5.0%) for treating bacterial corneal ulcers . METHODS: This randomized, parallel group, double-masked, multicenter study was conducted in 324 patients at 28 centers in the United States, Europe, and India . Patients were randomized into 2 treatment groups: 160 to ciprofloxacin and 164 to fortified tobramycin-cefazolin . Positive microbiologic cultures were obtained in 188 (58%) of 324 patients . Of these, 176 patients met protocol criteria and were evaluated for treatment efficacy: 82 in the ciprofloxacin group and 94 in the standard therapy group . The dosing schedule for both treatment groups was 1 to 2 drops of the first study medication (ciprofloxacin or fortified tobramycin) every 30 minutes for 6 hours, then hourly for the remainder of day 1; 1 to 2 drops every hour on days 2 and 3; 1 to 2 drops every 2 hours on days 4 and 5, followed by 1 to 2 drops every 4 hours on days 6 to 14 . The second medication (ciprofloxacin or cefazolin) was instilled 5 to 15 minutes after the first drug, following the same dosing frequency . Physician's judgment of clinical success, cure rate, changes in ocular sings, and symptoms and the rate of treatment failures were the primary efficacy criteria . RESULTS: Topical ciprofloxacin monotherapy is equivalent clinically and statistically to the standard therapy regimen of fortified antibiotics . No statistically significant treatment differences were found between ciprofloxacin (91.5%) and standard therapy (86.2%) in terms of overall clinical efficacy (P = 0.34) . Similarly, no differences were noted in resolution of the clinical signs and symptoms (P > 0.08) or the time to cure (P = 0.55) . The incidence of treatment failures was less in the ciprofloxacin group (8.5%) compared with the standard therapy group (13.8%) . Significantly fewer patients treated with ciprofloxacin reported discomfort than did patients treated with the standard therapy regimen (P = 0.01) . CONCLUSION: Ciprofloxacin ophthalmic solution 0.3% monotherapy is equivalent clinically and statistically to standard therapy (fortified tobramycin-cefazolin) for the treatment of bacterial corneal ulcers and produces significantly less discomfort. Gastrointest Endosc, 1996 Nov, 44(5), 583 - 6 Deficiencies of automatic endoscopic reprocessors: a method to achieve high-grade disinfection of endoscopes; Ido K et al.; BACKGROUND: We show that disinfection using the automatic endoscopic reprocessor is not complete and propose a method for high-grade disinfection of endoscopes . METHODS: We used an automatic endoscopic reprocessor, Pyser System 83, and 2% glutaraldehyde . After each endoscopic procedure, the endoscopes were divided into three groups . Endoscopes in group A were washed only by the reprocessor . Group B endoscopes were washed by the reprocessor after the connectors were soaked in glutaraldehyde for 5 minutes . The channels, valves, connecting sections of group C endoscopes, and the connectors of the machine were sprayed with glutaraldehyde before machine-washing . Swabs were taken from all 13 parts of each endoscope and machine for microbiologic culture . RESULTS: Six endoscopes were positive, cumulatively, for bacterial contamination in group A . Among group B endoscopes, one remained contaminated . No endoscope was positive in group C . The difference between group A and C was statistically significant (p < .05) . CONCLUSIONS: Machine washing by automatic endoscopic reprocessors may not achieve complete disinfection . Additional procedures are necessary . High-grade disinfection of the connectors is critical . Disinfection of the interface between the connectors is important. Clin Nucl Med, 1996 Nov, 21(11), 838 - 43 Tc-99m stannous colloid-labeled leukocyte scintigraphy in the evaluation of the painful arthroplasty; Chik KK et al.; The aim of this study was to evaluate the utility of scintigraphy using leukocytes labeled with Tc-99m stannous colloid in the assessment of painful arthroplasties . The results of three-phase bone scintigraphy and imaging with Tc-99m-labeled leukocytes in 40 patients with painful joint prostheses, requiring the exclusion of infection are reported . Results were compared with microbiologic culture and/or long-term clinical review . Bone scintigraphy had a sensitivity of 100%, a specificity of 64%, and a diagnostic accuracy of 73% in detecting prosthetic infection . Tc-99m-labeled leukocytes yielded a sensitivity of 70%, a specificity of 100%, and a diagnostic accuracy of 93% . Leukocyte scintigraphy improves the specificity of bone scintigraphy in the assessment of the painful prosthetic joint . A positive study is highly suggestive of infection and warrants appropriate treatment. Ann Pharmacother, 1996 Nov, 30(11), 1238 - 42 Sterility of filgrastim (G-CSF) in syringes; Jacobson PA et al.; OBJECTIVE: To determine if the sterility of filgrastim (G-CSF) is maintained for up to 7 days when aseptically transferred from the vial to tuberculin syringes in a laminar air flow environment . DESIGN: The study was conducted in two phases: a validation and an experimental phase . The method was validated by inoculating samples of sterile filgrastim solution with common bacterial and fungal skin contaminants . Samples were aseptically drawn into syringes in a class 100 horizontal laminar air flow hood and refrigerated . The samples were equally divided and transferred to microbiology culture media at times 0, 24 hours, 48 hours, and 7 days; incubated; and the organisms identified and quantitated . In the experimental phase, samples of filgrastim were aseptically drawn into syringes, separated into three groups, and refrigerated . At 24 hours, 48 hours, and 7 days, the samples were transferred to broth, incubated, and observed for the development of turbidity . SETTING: A class 100 laminar air flow hood in a pediatric hospital pharmacy and a home-infusion pharmacy class 100,000 clean room . MAIN OUTCOME MEASURES: The sterility of filgrastim in syringes was determined by comparing experimental broth culture tubes to a control tube and observing for the development of turbidity . RESULTS: Filgrastim demonstrated the ability to support the growth of intentionally inoculated skin contaminants, both qualitatively and quantitatively . However, when aseptically transferred to syringes and refrigerated, all tested filgrastim samples remained sterile for at least 7 days . CONCLUSIONS: Syringes of filgrastim remain sterile for 7 days when prepared in a class 100 laminar air flow hood, using aseptic technique, and stored under refrigeration . This change in practice can result in significant cost savings. Infect Immun, 1996 Nov, 64(11), 4694 - 9 No apparent influence of immunoglobulins on indigenous oral and intestinal microbiota of mice; Marcotte H et al.; The role of secretory immunoglobulin A (sIgA) in the control of the indigenous microbiota is not well understood . In this study, we compared the oral and intestinal microbiota of transgenic B-cell-deficient (microMT) mice with their heterozygous (microMT/+) normal littermates . The levels of salivary IgA and serum IgA and IgG were normal in microMT/+ mice, while no immunoglobulins were detected in microMT/microMT mice . The acquisition and proportions of the different species of the oral and intestinal indigenous bacterial populations were not significantly different between the two groups of mice . Our results thus suggest that secretory IgA does not play a major role in the regulation of the indigenous microbiota of mice. Gene, 1996 Oct 24, 177(1-2), 243 - 51 Characterization of spaA, a Streptomyces coelicolor gene homologous to a gene involved in sensing starvation in Escherichia coli; Schneider D et al.; A Streptomyces coelicolor gene, called spaA, homologous to the stationary phase regulatory gene rspA of Escherichia coli {Huisman and Kolter (1994) Science 265, 537-539}, was cloned using the Streptomyces ambofaciens rspA homologue spa2 {Schneider et al . (1993) J . Gen . Microbiol . 139, 2559-2567} as a probe . Considerable differences in sequence and in genetic context were detected between spa2 of S . ambofaciens and spaA of S . coelicolor . A cloned internal fragment of spaA was used to direct integration of a phage vector into the spaA gene . The disruption caused delayed antibiotic production (undecylprodigiosin and actinorhodin) and led on further incubation to increased actinorhodin production at high, but not low, cell density . This phenotype was apparent only on the nutritionally poorest of three media tested . The attempted use of an integrating plasmid-based system for gene replacement of spaA gave rise to extensive deletions of adjacent chromosomal DNA. FEMS Microbiol Lett, 1996 Oct 15, 144(1), 109 - 15 Enhancement of frame-shift mutation by the overproduction of msDNA in Escherichia coli; Mao JR et al.; A minor population of wild Escherichia coli strains contain retroelements called retrons, which produce a peculiar satellite DNA, multicopy single-stranded DNA (msDNA) . It has been reported that mismatched base pairs in the secondary structure formed in msDNA are mutagenic in E . coli{Maas et al.(1994) Mol.Microbiol . 14,437-441; Maas et al . (1996) Mol . Microbiol, 19, 505-509} . We reexamined this proposal by converting mismatched base pairs to matched base pairs using a single msDNA species, msDNA-Ec86, or by deleting mismatched regions using msDNA-Ec73 . We also examined the effect of reverse transcriptases (RT) without msDNA production on mutagenesis . All the constructs are under the lpp/lac promoter-operator control so that their mutagenic effects can be tested in the absence and the presence of a lac inducer . It was found that when the production of msDNA-Ec86 or Ec73 was induced, reversion frequencies from Lac- to Lac+ significantly increased in the case of a Lac- mutation caused by a frame-shift mutation, but much less by a substitution mutation . The removal of mismatched base pairs eliminated the high mutation frequencies, and the inducible expression of RT alone was not mutagenic . These results are consistent with the hypothesis of Maas and his associates that mismatched base pairs in msDNA sequester a cellular mismatch repair system, resulting in the increase of frame-shift mutations. Ultraschall Med, 1996 Oct, 17(5), 253 - 6 {Tuberculoid caseating necrosis of cervical lymph nodes . Ultrasound after radiochemotherapy of undifferentiated nasopharyngeal carcinoma}; Friedrich RE et al.; Sonography following Radiochemotherapy for Undifferentiated Nasopharyngeal Carcinoma: AIM AND METHOD: An undifferentiated nasopharyngeal carcinoma (UCNT) recurred 5 1/2 months after radiochemotherapy in a 14-year-old Caucasian boy . By ultrasound, multiple lymph nodes, predominantly left sided, were identified both in groups and singly located adjacent to large cervical vessels . RESULTS: After lymphadenectomy, pathological examination revealed caseation necrosis with epitheloid cells and Langhans {corrected} cells in 2 of 6 enlarged lymph nodes (> 10 mm) . The tentative diagnosis of tuberculosis was excluded during (3 years) follow-up . CONCLUSION: Caseation necrosis in lymph node metastases is highly characteristic of UCNT . Sonography failed in the case described to demonstrate differences in lymph nodes with and without caseation necrosis . Differential diagnosis between tuberculous and metastatic cervical lymph nodes must therefore be made by other means (serology, microbiology). Pneumologie, 1996 Oct, 50(10), 712 - 7 {Community-acquired pneumonia requiring inpatient treatment: outcome and prognostic rule}; Ewig S et al.; OBJECTIVE: Mortality in community-acquired pneumonia (CAP) may be reduced by early identification of patients requiring intensive care treatment . The purpose of the study was to determine prognostic factors of outcome in patients with CAP in order to establish a clinically applicable discriminant rule . METHODS: 93 episodes of 92 patients with CAP were reviewed with regard to epidemiological, clinical, laboratory and microbiologic data . The prognostic analysis included a univariate as well as a multivariate approach in order to identify parameters correlated with death using the Cox regression hazard function in a backward stepwise selection model . The three parameters found to contribute most to the significance of the model were used in a discriminant rule for classification of outcome . RESULTS: The parameters found to be significantly different between survivors and non-survivors were heart rate, systolic, diastolic as well as mean blood pressures, leucocyte count, percentage of laymphocytes, and LDH values . The multivariate analysis revealed that heart rate, systolic arterial pressure, and LDH serum levels were associated best with lethal outcome (overall significance of the model p < 0.005) . A prognostic rule composed of the variables heart rate > or = 90 beats/min, systolic arterial blood pressure < or = 80 mmHg and LDH > or = 260 U/l achieved a sensitivity of 77%, a specificity of 75% and positive and negative predictive values of 42% and 93%, respectively . It was associated with a 6-fold increased risk of lethal outcome . CONCLUSIONS: Heart rate, systolic blood pressure, and LDH values were associated best with death in a multivariate analysis . A discriminant rule consisting of these three variables achieved favourable classification results . The rule qualifies for further prospective validation and may prove useful in the management of hospital treated CAP. Heart, 1996 Oct, 76(4), 337 - 43 Specific changes in skeletal muscle myosin heavy chain composition in cardiac failure: differences compared with disuse atrophy as assessed on microbiopsies by high resolution electrophoresis; Vescovo G et al.; OBJECTIVE: In congestive heart failure (CHF) the skeletal muscle of the lower limbs develops a myopathy with atrophy and shift from the slow type to the fast type fibres . The aim was to test the hypothesis that this myopathy is specific and not simply related to detraining, by comparing patients with different degrees of CHF with patients with severe muscle atrophy due to disuse . DESIGN: Case-control study involving 50-150 micrograms needle biopsies of the gastrocnemius muscle . By an electrophoretic micromethod, the three isoforms of myosin heavy chains (MHC) were separated . PATIENTS: Five patients restricted to bed for more than one year because of stroke with disuse atrophy and normal ventricular function, and 19 with CHF were studied . There were seven age matched controls . MAIN OUTCOME MEASURES: The percentage of MHC1 (slow isoform), MHC2a (fast oxidative), and MHC2b (fast glycolytic) was determined by densitometric scan and correlated with indices of severity of cardiac failure . RESULTS: Ejection fraction was 42.5 (SD 15.2)% in CHF, 59.5 (1.0)% in disuse atrophy and 60.3 (1.4)% in controls (P < 0.001 v both) . The degree of muscle atrophy as calculated by the body mass index/gastrocnemius cross sectional area, showed a profound degree of atrophy in patients with muscle disuse {0.94 (0.39)} . This was worse than in the controls {4.27 (0.16), P < 0.0005} and the CHF patients {2.60 (1.10), P < 0.005} . Atrophy in CHF patients was also greater than in controls (P < 0.005) . MHC1 was lower in CHF than in disuse atrophy {51.83 (15.04) v 84.5 (17.04), P < 0.01} while MHC2b was higher {23.5 (7.4) v 7.25 (7.92), P < 0.001} . There was a similar trend for MHC2a {24.83 (15.01) v 8.25 (9.12), P < 0.05} . Within the CHF group there was a positive correlation between NYHA class and MHC2a (r = 0.47, P < 0.05) and MHC2b (r = 0.55, P < 0.01) and a negative correlation between NYHA class and MHC1 (r = -0.74, P < 0.001) . Similarly, significant correlations were found for ejection fraction, diuretic consumption score, exercise test tolerance, and degree of muscle atrophy . CONCLUSIONS: The CHF myopathy appears to be specific and not related to detraining . The magnitude of MCH redistribution correlates with the severity of the disease . The electrophoretic micromethod used is very sensitive and reproducible . Biopsies are so well tolerated that can be repeated frequently, allowing thorough follow up. Pediatr Neurol, 1996 Oct, 15(3), 230 - 4 Primary amoebic meningoencephalitis with Naegleria fowleri: clinical review; Barnett ND et al.; Two children with primary amoebic meningoencephalitis secondary to Naegleria fowleri are reported . Both children died, and the causative agent was identified at autopsy . Presentation and outcome conformed to the usual course of primary amoebic meningoencephalitis and reaffirm the gravity and rapid progression of this infection . The epidemiology, microbiology, diagnostic considerations, and treatment are discussed . Primary amoebic meningoencephalitis should be considered in the differential diagnosis of children with meningitis or encephalitis. Semin Perinatol, 1996 Oct, 20(5), 409 - 17 The evaluation of infection and pulmonary maturity in women with premature rupture of the membranes; Beazley D et al.; Although the etiology of PROM is multifactorial, increasing evidence regarding clinical risk factors, membrane histology, membrane culture, and amniotic fluid microbiology shows a strong association with infection . Recent studies suggest an association between genital tract infection, preterm labor (PTL), and preterm premature rupture of the membranes (pPROM) . If correct, this information may be used to target areas for prevention, as well as to develop management protocols . This article reviews both the diagnostic tests for the causes associated with pPROM and the evaluation of intraamniotic infection and pulmonary maturity in patients with pPROM. Infect Control Hosp Epidemiol, 1996 Oct, 17(10), 641 - 8 Prevention of nosocomial influenza; Adal KA et al.; OBJECTIVE: To study compliance with preventive strategies at a university hospital during an outbreak of nosocomial influenza A during the winter of 1988, and the rates of vaccination of healthcare workers and of nosocomial influenza following changes in vaccine practices after the outbreak . DESIGN: Retrospective review of employee health, hospital epidemiology, hospital computing; and clinical microbiology records . SETTING: A university hospital . INTERVENTIONS: Unvaccinated personnel with exposure within the previous 72 hours to an unisolated case of influenza were offered influenza vaccine and 14 days of amantadine hydrochloride prophylaxis . Personnel with exposure more than 72 hours before evaluation were offered vaccine . A mobile cart was introduced for vaccinating personnel after the 1988 outbreak . RESULTS: An outbreak of influenza with 10 nosocomial cases occurred in 1988 . Only 4% of exposed employees had been vaccinated previously and 23% of exposed, unvaccinated employees agreed to take vaccine, amantadine, or both . A mobile-cart vaccination program was instituted, and annual vaccination rates steadily increased from 26.3% in 1989 to 1990 to 38% in 1993 to 1994 (P < .0001) . The relative frequency of documented cases of influenza in employees with symptoms of influenza decreased significantly during this period (P = .025), but nosocomial influenza rates among patients did not change significantly . CONCLUSION: A mobile-cart influenza vaccination program was associated with a significant increase in compliance among healthcare workers, but a majority still remained unvaccinated . The rate of nosocomial influenza among patients was not reduced by the modest increase in the vaccination rate, but influenza rates remained acceptably low, perhaps due to respiratory isolation of patients and furlough of employees with influenza. Clin Orthop, 1996 Oct, (331), 11 - 22 The Coventry Award . Polymerase chain reaction detection of bacterial infection in total knee arthroplasty; Mariani BD et al.; Synovial fluid aspirates from 50 patients with symptoms after total knee arthroplasty were analyzed by means of the polymerase chain reaction for the presence of bacterial deoxyribonucleic acid indicative of infection . Synovial fluid specimens were processed using a rapid bacterial lysis and extraction protocol, subjected to polymerase chain reaction amplification using universal bacterial primers, and polymerase chain reaction products analyzed by deoxyribonucleic acid hybridization methodology . Polymerase chain reaction testing on preoperative aspirates yielded 32 specimens positive for bacterial infection . Standard microbiologic culturing assays performed on the same samples gave 6 positive bacterial infection tests; intraoperative culturing identified 9 additional infected specimens . All culture positive specimens were polymerase chain reaction positive; in contrast, there were no false polymerase chain reaction positives in 21 negative control specimens obtained from aseptic joints . The synovial fluid processing protocol and polymerase chain reaction analysis can be performed with a minimum of time and may provide greater sensitivity than standard diagnostic tests . In view of the high incidence of false negative test results from standard microbiologic assays of synovial fluid specimens, the use of molecular biology based bacterial detection methodology should provide an additional, or alternative, assay to identify infected patient specimens. Clin Microbiol Rev, 1996 Oct, 9(4), 435 - 47 Mycobacterium haemophilum: microbiology and expanding clinical and geographic spectra of disease in humans; Saubolle MA et al.; Reports of the association of Mycobacterium haemophilum with disease in humans have greatly increased . At least 64 cases have now been reported, with symptoms ranging from focal lesions to widespread, systemic disease . The organism is now known to cause primarily cutaneous and subcutaneous infection, septic arthritis, osteomyelitis, and pneumonitis in patients who are immunologically compromised and lymphadenitis in apparently immunocompetent children . Underlying conditions in the compromised patients have included AIDS; renal, bone marrow, and cardiac transplantation; lymphoma; rheumatoid arthritis; marrow hypoplasia; and Crohn's disease . Reports have originated from diverse geographic areas worldwide . The epidemiology of M . haemophilum remains poorly defined; there appears to be a genetic diversity between strains isolated from different regions . The organism is probably present in the environment, but recovery by sampling has not been successful . M . haemophilum has several unique traits, including predilection for lower temperatures (30 to 32 degrees C) and requirement for iron supplementation (ferric ammonium citrate or hemin) . These may in the past have compromised recovery in the laboratory . Therapy has not been well elucidated, and the outcome appears to be influenced by the patient's underlying immunosuppression . The organisms are most susceptible to ciprofloxacin, clarithromycin, rifabutin, and rifampin . Timely diagnosis and therapy require communication between clinician and the laboratory. Otolaryngol Clin North Am, 1996 Oct, 29(5), 761 - 82 Otitis externa; Bojrab DI et al.; Otitis externa is a broad term for a disease state that includes inflammation or infection of the external auditory canal and auricle . It can range from mild inflammation and discomfort to a life-threatening disease . Treatment of otitis externa is dependent on a thorough understanding of anatomy and physiology of the external ear canal, a knowledge of the microbiology of potential pathogens, and familiarity with clinical presentation, so that an accurate and timely diagnosis can be reached. Ann Pharmacother, 1996 Oct, 30(10), 1115 - 6 Meningitis and hepatitis complicating intravenous immunoglobulin therapy; Shorr AF et al.; OBJECTIVE: To report a case of concomitant meningitis and hepatitis complicating the use of intravenous immune globulin (IVIG) . CASE SUMMARY: A 39-year-old African-American woman with an autoimmune syndrome developed both acute meningitis and hepatitis following administration of IVIG . These resolved over several days and left no sequellae . DISCUSSION: This represents the first case of concomitant acute meningitis and hepatitis associated with IVIG . Thorough microbiologic and serologic evaluation of the patient failed to demonstrate an infectious etiology . We postulate that our patient's syndrome resulted from direct toxicity of IVIG . CONCLUSIONS: Both acute meningitis and hepatitis may simultaneously complicate IVIG therapy . The specific mechanism remains unclear. J Clin Microbiol, 1996 Oct, 34(10), 2604 - 6 Comparison of reverse transcription-PCR with tissue culture and other rapid diagnostic assays for detection of type A influenza virus; Atmar RL et al.; We applied a reverse transcription (RT)-PCR assay for influenza A virus to combined nasal wash-throat swab specimens previously obtained from an outpatient pediatric population with acute respiratory illness during concurrent epidemics of influenza A virus and respiratory syncytial virus . The results of the RT-PCR assay were compared with those previously reported with virus cultivation and commercially available rapid diagnostic kits (E.A . Dominguez, L.H . Taber, and R.B . Couch, J . Clin . Microbiol . 31:2286-2290, 1993) . With virus cultivation as the "gold standard", the RT-PCR assay had a sensitivity, specificity, and efficiency of 95, 98, and 97%, respectively, compared with 75, 100, and 93%, respectively, for the best diagnostic kit (Becton Dickinson Directigen) . RT-PCR is an effective alternative to virus isolation for the detection of influenza A virus in clinical specimens. J Clin Microbiol, 1996 Oct, 34(10), 2469 - 74 Differentiation of Mycobacterium bovis isolates from animals by DNA typing; Skuce RA et al.; The insertion sequence IS6110 and the direct repeat (DR) specific to tuberculosis complex mycobacteria and the highly repeated DNA sequence, the polymorphic GC-rich repeat sequence (PGRS), were systematically used to identify restriction fragment length polymorphisms (RFLPs) within 210 isolates of Mycobacterium bovis . The isolates were primarily of bovine origin, but isolates from badgers, feral deer, sheep, humans, and a pig were included . The RFLP probes IS6110, DR, and PGRS individually identified 17, 18, and 18 different RFLP types, respectively, but in combination these probes identified a total of 39 different M . bovis RFLP types . The recommendations (J . D . A . van Embden, M . D . Cave, J . T . Crawford, J . W . Dale, K . D . Eisenach, B . Gicquel, P . W . M . Hermans, C . Martin, R . McAdam, T . M . Shinnick, and P . M . Small, J . Clin . Microbiol . 31:406-409, 1993) for a standardized RFLP analysis for M . tuberculosis were adapted to facilitate gel documentation, image analysis, and construction of a database of RFLP types . In the present study the same M . bovis RFLP types were evident in the various animal species included, indicating that the strains were not host restricted . Application of these techniques to defined field studies should help elucidate more accurately aspects of the epidemiology of bovine tuberculosis in different countries. Int J Syst Bacteriol, 1996 Oct, 46(4), 933 - 8 A proposal to transfer Microbispora bispora (Lechevalier 1965) to a new genus, Thermobispora gen . nov., as Thermobispora bispora comb . nov; Wang Y et al.; We determined almost complete 16S rRNA gene sequences of two Microbispora bispora (Lechevalier 1965) strains, ATCC 19993T (T = type strain) and JCM 3082 . The two sequences were 99% similar to each other but exhibited only 81 to 87.8% similarity with the 16S rRNA gene sequences of seven other Microbispora strains . A phylogenetic analysis revealed that the two sequences clustered not only distantly from other Microbispora strains, but also outside the cluster containing members of the family Streptosporangiaceae . On the basis of the results of our phylogenetic analysis and the results of a comprehensive review of the genus Microbispora by Miyadoh et al . (S . Miyadoh, S . Amano, H . Tohyama, and T . Shomura, J . Gen . Microbiol . 136:1905-1913, 1990) in which chemotaxonomic and DNA-DNA hybridization analyses were performed, we propose that Microbispora bispora should be transferred to a new genus, Thermobispora gen . nov., as Thermobispora bispora comb . nov. J Trauma, 1996 Oct, 41(4), 703 - 7 Comparison of nonbronchoscopic techniques with bronchoscopic brushing in the diagnosis of ventilator-associated pneumonia; Wearden PD et al.; OBJECTIVE: To determine the diagnostic accuracy of quantitative cultures obtained via nonbronchoscopic protected specimen brushing (PSB) and nonbronchoscopic bronchoalveolar lavage (BAL) compared with quantitative cultures obtained by bronchoscopic PSB in surgical patients suspected of ventilator-associated pneumonia . DESIGN: Prospective, crossover controlled study of 15 ventilated surgical intensive care unit patients in a university teaching hospital . METHODS: Fifteen consecutive ventilated patients suspected of ventilator-associated pneumonia on the basis of leukocytosis, purulent sputum, and appearance of chest roentgenogram were enrolled . All patients underwent nonbronchoscopic PSB and BAL followed by bronchoscopic PSB . The duration of each procedure was noted . Culture results were considered positive only if greater than 10(4) colony forming units (CFU) per milliliter were present . MAIN RESULTS: Perfect concordance was noted between bronchoscopic PSB and nonbronchoscopic BAL (kappa = 1.0) . The concordance between bronchoscopic PSB and nonbronchoscopic PSB was 93% ((kappa = 0.86) . The nonbronchoscopic procedures were performed in significantly less time than the bronchoscopic procedure . CONCLUSION: Nonbronchoscopic PSB and BAL provide similar microbiologic data to bronchoscopic PSB in the diagnosis of ventilator-associated pneumonia while shortening procedure time significantly. Nat Med, 1996 Oct, 2(10), 1116 - 21 Oral immunization with an anti-idiotypic antibody to the exoglycolipid antigen protects against experimental Chlamydia trachomatis infection; Whittum-Hudson JA et al.; Chlamydia trachomatis is the leading cause worldwide of preventable infectious blindness (trachoma) and sexually transmitted disease, including nongonoccocal urethritis and pelvic inflammatory disease . To date, no effective vaccine against C . trachomatis infection has been identified . A monoclonal anti-idiotypic antibody (anti-Id) to the chlamydial exoglycolipid antigen (GLXA) was tested in a murine model of ocular chlamydial infection for its ability to induce systemic immunity, which reduces microbiologic and clinical disease . The anti-Id to GLXA, delivered either systemically in soluble form or orally after encapsulation in poly(lactide) microspheres, induced significant protective immunity against ocular challenge of mice with a human biovar of C . trachomatis . Protection was associated with induction of anti-GLXA antibody and anti-chlamydial neutralizing antibody. Appl Environ Microbiol, 1996 Oct, 62(10), 3861 - 3 Improved method for the preparative synthesis of labeled trehalose of high specific activity by Escherichia coli; Horlacher R et al.; We report an improvement of a published procedure using Escherichia coli to synthesize 14C-labeled trehalose from {14C}glucose (B . Brand and W . Boos, Appl . Environ . Microbiol . 55:2414-2415, 1989) . Instead of inducing the expression of the trehalose-synthesizing enzymes encoded by the chromosomal genes otsAB by high osmolarity, we now induce their expression from a plasmid under normal growth conditions by the addition of IPTG (isopropyl-beta-D-thiogalactopyranoside) . Instead of using a pgi zwf double mutant to prevent glucose utilization, we use a pgi::Tn10 insertion only . In addition to being defective in treA, which encodes a periplasmic trehalase, the strain is now also defective in treF, which encodes a newly discovered cytoplasmic trehalase . This strain is genetically stable; it has no growth defects; and after induction with IPTG, it will transform {14C}glucose to {14C}trehalose in minimal medium without any carbon source under aerobic conditions at a rate of 3 nmol/min/10(9) cells . With the improved method, the overall yield of trehalose from glucose is about 80% and the process takes place without dilution of the specific radioactivity of the glucose residues . The accumulated trehalose is extracted from the bacteria by 70% hot ethanol and can easily be purified radiochemically by chromatographic techniques. J Bacteriol, 1996 Oct, 178(19), 5847 - 9 The cbpA chaperone gene function compensates for dnaJ in lambda plasmid replication during amino acid starvation of Escherichia coli; Wegrzyn A et al.; We found previously that lambda plasmid DNA replication in amino acid-starved Escherichia coli relA mutants (i.e., during the relaxed response), which is carried out by the inherited replication complex, is dependent on functions of DnaK and GrpE molecular chaperones but proceeds in a dnaj mutant at a nonpermissive temperature . Here we demonstrate that this replication is inhibited when functions of both dnaJ and cbpA are impaired . In complete media, the growth of the lambda pi A66 phage (capable of replicating in E . coli dnaJ, dnaK, and grpE missense mutants at 30 degrees C), as well as efficiency of transformation by the lambda pi A66 plasmid, is significantly decreased in a dnaJ259 cbpA::kan double mutant . These results strengthen the proposal of other authors (C . Ueguchi, M . Kakeda, H . Yamada, and T . Mizuno, Proc . Natl . Acad . Sci . USA 91:1054-1058, 1994; C . Ueguchi, T . Shiozawa, M . Kakeda, H . Yamada, and T . Mizuno, J . Bacteriol . 177:3894-3896, 1995; and T . Yamashino, M . Kakeda, C . Ueguchi, and T . Mizuno, Mol . Microbiol . 13:475-483, 1994) that the cbpA gene product is a functional analog of the DnaJ chaperone in E . coli. J Virol, 1996 Oct, 70(10), 6973 - 81 The rotavirus nonstructural glycoprotein NSP4 possesses membrane destabilization activity; Tian P et al.; During a unique morphogenetic process, rotaviruses obtain a transient membrane envelope when newly synthesized subviral particles bud into the endoplasmic reticulum (ER) . As rotavirus particles mature, they lose their transient membrane and a layer of the glycoprotein VP7 forms the virion outer capsid shell . The nonstructural glycoprotein NSP4 functions as an intracellular receptor in the ER membrane (K . S . Au, W . K . Chan, J . W . Burns, and M . K . Estes, J . Virol . 63:4553-4562, 1989), and it has been hypothesized that NSP4 is involved in the removal of the envelope during viral morphogenesis (M . K . Estes and J . Cohen, Microbiol . Rev . 53:410-449, 1989; B . L . Petrie, M . K . Estes, and D . Y . Graham, J . Virol . 46:270-274, 1983) . The purpose of the present study was to determine if NSP4 has a direct membrane destabilization activity (MDA) by using liposome leakage assays and electron microscopic visualization of liposome, microsome, and viral envelope disruption . The fluorescent marker (calcein) incorporated into liposomes was released when the liposomes were incubated with purified NSP4 . A region corresponding to amino acid residues 114 to 135 of NSP4 also released calcein from liposomes . NSP4(114-135) peptide-specific antibody completely blocked the MDA of the purified NSP4 protein . These results suggest that this region contains at least part of the functional domain of NSP4 . Liposomes composed of phosphatidylcholine and microsomes (to simulate ER membranes) were broken when observed by electron microscopy after incubation with NSP4 or the NSP4(114-135) peptide . In contrast, the envelope of Sendai virus, which is derived from cytoplasmic membranes, and erythrocytes were not disrupted by NSP4 and the NSP4(114-135) peptide . These results provide direct evidence that NSP4 possesses MDA and suggest that it can cause ER membrane damage . Therefore, NSP4 might play an important role in the removal of the transient envelope from budding particles during viral morphogenesis . A model for the MDA of NSP4 in viral morphogenesis is proposed. FEBS Lett, 1996 Sep 23, 394(1), 14 - 6 Purification and characterization of the Escherichia coli thermoresistant glucokinase encoded by the gntK gene; Izu H et al.; A thermoresistant gluconokinase encoded by the gntK gene of Escherichia coli K-12 was purified and characterized . The Km values of the purified enzyme for gluconate and ATP are 42 microM and 123 microM, respectively, and the activity was not altered by the presence of pyruvate . The enzyme was shown to function as a dimer with two identical subunits of 18.4 kDa . These characteristics appear to be distinct from those of the gluconokinase reported by E.I . Vivas, A . Liendo, K . Dawidowicz, and T . Isturiz (1994) J . Basic . Microbiol . 16, 117-122. Eur J Biochem, 1996 Sep 15, 240(3), 491 - 507 The regulation of human immunodeficiency virus type-1 gene expression; Kingsman SM et al.; Despite 15 years of intensive research we still do not have an effective treatment for AIDS, the disease caused by human immunodeficiency virus (HIV) . Recent research is, however, revealing some of the secrets of the replication cycle of this complex retrovirus, and this may lead to the development of novel antiviral compounds . In particular the virus uses strategies for gene expression that seem to be unique in the eukaryotic world . These involve the use of virally encoded regulatory proteins that mediate their effects through interactions with specific viral target sequences present in the messenger RNA rather than in the proviral DNA . If there are no cellular counterparts of these RNA-dependent gene-regulation pathways then they offer excellent targets for the development of antiviral compounds . The viral promoter is also subject to complex regulation by combinations of cellular factors that may be functional in different cell types and at different cell states . Selective interference of specific cellular factors may also provide a route to inhibiting viral replication without disrupting normal cellular functions . The aim of this review is to discuss the regulation of HIV-1 gene expression and, as far as it is possible, to relate the observations to viral pathogenesis . Some areas of research into the regulation of HIV-1 replication have generated controversy and rather than rehearsing this controversy we have imposed our own bias on the field . To redress the balance and to give a broader view of HIV-1 replication and pathogenesis we refer you to a number of excellent reviews {Cullen, B . R . (1992) Microbiol . Rev . 56, 375-394; Levy, J . A . (1993) Microbiol . Rev . 57, 183-394; Antoni, B . A., Stein, S . & Rabson, A . B . (1994) Adv . Virus Res . 43, 53-145; Rosen, C . A . & Fenyoe, E . M . (1995) AIDS (Phila.) 9, S1-S3}. Anal Chem, 1996 Sep 15, 68(18), 3173 - 9 Thin-film microbiosensors for glucose-lactate monitoring; Jobst G et al.; A miniaturized device for simultaneous measurement of glucose and lactate levels was produced by means of photopatterning of enzyme-containing photosensitive membrane precursors . This device shows no cross-talk and a lifetime for both the glucose and the lactate sensors of more than 2 weeks when continuously operated in undiluted bovine serum . Linear response ranges of up to 40 mM for glucose and 25 mM for L-lactate, in combination with 95% response times of < 30 s, were realized . The devices are mass produced by means of thin-film technology on flexible carriers to give catheter-type multisensing devices for in vivo applications . Ex vivo experiments, performed with human volunteers, where the device was continuously operated in an extracorporeal, undiluted, heparinized blood stream for 6 h, gave a correlation of r > 0.98 with respect to laboratory techniques . Subcutaneous measurements of glucose levels in pigs were close to the corresponding blood levels obtained without in vivo calibration. Commun Dis Rep CDR Rev, 1996 Sep 13, 6(10), R144 - 6 PHLS begins a national case control study of Escherichia coli O157 infection in England; Adak GK et al.; Infection with Vero cytotoxin producing Escherichia coli O157 (VTEC O157) is a growing public health problem and the commonest cause of acute renal failure in children in the United Kingdom . Foodborne outbreaks of VTEC O157 infection have been reported in the United Kingdom, other European countries, and North America . Most cases of infection are sporadic, however, and the contribution of food vehicles, animal contact, and person to person spread in the acquisition of infection needs to be clarified . The PHLS is starting a case control study in England to identify and estimate the relative importance of risk factors for the acquisition of VTEC O157 infection . The study will run for 12 months . This article describes its objectives and asks microbiologists, public health physicians, clinicians, and others who may be asked for details about cases or to find suitable controls for their help in achieving a successful outcome. Methods Inf Med, 1996 Sep, 35(3), 189 - 92 Medical informatics: once more towards systematization; Haux R; Commenting on a paper by Van Bemmel (Medical Informatics, Art or Science? {1}), the following questions are raised: What is the meaning of medical informatics?, How to systematize medical informatics?, is medical informatics an art, a science or a technology? . It is argued that medical informatics is concerned with the systematic processing of data, information and knowledge in medicine and health care, and that medical informatics is not just the application of computers in these fields . Three classifications for medical informatics research and education are presented . It is concluded that medical informatics is a scientific medical discipline, similar to surgery, internal medicine, epidemiology, or microbiology; and that medical informatics has a strong relationship with the health sciences concerning its field of application, and to informatics concerning its methods and tools . It is a cross-sectional discipline, with relevance for virtually all other specialties of medicine and the health sciences . This is the reason for its impact on research and education in these specialties . It also causes that the quality of the processing of data, information and knowledge has a direct and considerable effect upon the quality of health care in practically all these specialties. Chem Biol, 1996 Sep, 3(9), 757 - 63 Optically controlled collisions of biological objects to evaluate potent polyvalent inhibitors of virus-cell adhesion; Mammen M et al.; BACKGROUND: The biochemical and biomechanical determinants of adhesion between two biological objects following a