Lipids, 1996 Jan, 31(1), 53 - 9 Retinal fatty acids of piglets fed docosahexaenoic and arachidonic acids from microbial sources; Craig-Schmidt MC et al.; Docosahexaenoic acid (DHA, 22:6n-3) and arachidonic acid (AA, 20:4n-6) serve important roles in perinatal visual and neural development . A neonatal pig model was used to determine if dietary supplementation with DHA and AA at slightly greater concentrations than normally found in human milk would influence fatty acid accretion in retina . One-day-old piglets were assigned to one of four diets (n = 5/group): (i) STD, standard diet containing fat similar to infant formula; (ii) STD + DHA, 0.7% of fatty acids as DHA; (iii) STD + AA, 0.9% as AA; and (iv) STD + BOTH, 0.8% as DHA plus 1.0% as AA . After 25 d, fatty acids in retina phosphatidylcholine (PC) and phosphatidylethanolamine (PE) were determined . Supplementation with DHA resulted in approximately twofold increases (P < 0.05) in PC-DHA (4.88% in STD vs . 10.03% in STD + DHA and 9.47% in STD + BOTH) . Similarly, AA supplementation increased PC-AA 1.3-1.4-fold (4.47% in STD vs . 6.19% in STD + AA and 5.70% in STD + BOTH) . For PE, supplementation with either fatty acid or in combination resulted in no significant increases, except for a 1.2-fold increase in DHA for STD + BOTH (32.66%) vs . STD (28.38%) . Thus, PC responded to dietary supplementation, with addition of DHA, AA, or BOTH, resulting in increases in respective fatty acids; PE was less responsive, with only STD + BOTH resulting in increased DHA . No significant competition between DHA and AA in incorporation into phospholipids was observed . In conclusion, consumption of a combination of DHA and AA by neonatal pigs supported accretion of DHA in retina phospholipids, while simultaneously supplying the AA necessary for membrane phospholipids and eicosanoid biosynthesis. Arch Virol, 1996, 141(3-4), 743 - 9 Immunisation with DNA polynucleotides protects mice against lethal challenge with St . Louis encephalitis virus; Phillpotts RJ et al.; In vivo transfection by intramuscular injection with plasmids expressing the immunogenic proteins of microbial pathogens has considerable potential as a vaccination strategy against many pathogens of both man and animals . Here we report that weanling mice given a single intramuscular injection of 50 micrograms of a plasmid, pSLE1 expressing the St . Louis encephalitis virus (SLE) prM/E protein under the control of the cytomegalovirus immediate early protein promoter produced SLE-specific antibody and were protected against lethal challenge with the virulent virus . Polynucleotide vaccine technology provides a unique opportunity to produce vaccines against flavivirus diseases of low incidence cheaply and rapidly, and to produce multivalent vaccines such as would be required for immunisation against dengue virus disease. Ophthalmology, 1996 Jan, 103(1), 29 - 36 Limbal transplantation; Tan DT et al.; BACKGROUND: Limbal transplantation is a surgical technique of ocular surface epithelial transplantation advocated for a variety of ocular surface disorders with presumed stem-cell deficiency . Limbal transplantation was performed in 18 patients with ocular surface disease, which included aniridia keratopathy, chronic contact lens-associated epitheliopathy, chemical injury, Stevens-Johnson syndrome, and corneal intraepithelial dysplasia . METHODS: Limbal allograft transplantation was performed in nine eyes with the use of heterologous limbal tissue from cadaveric donor eyes or live relatives, whereas nine eyes underwent conventional limbal autograft transplantation . Results: Limbal allograft transplantation resulted in restoration of a stable ocular surface in seven of nine cases, with early visual rehabilitation and significant reduction in symptoms . At a mean follow-up period of 14.7 months, one patient was noted to have failure of the inferior graft related to postoperative microbial keratitis, whereas one patient had acute rejection episode after early cessation of oral cyclosporine . The mean follow-up period for limbal autografts was 27.1 months . Limbal autograft failure occurred in two patients with limbal autograft transplantation for chronic contact lens-associated epitheliopathy . One contact lens wearer had epithelial dysplasia in the fellow eye at the previous donor site . Subclinical involvement of the fellow eye is suggested as a reason for graft failure and donor eye complications in these eyes . CONCLUSIONS: Although it requires a longer follow-up period, limbal allograft transplantation is a viable procedure for bilateral ocular surface disease and for presumed bilateral disease (viz bilateral exposure to injurious agents) and may be a preferred alternative to limbal autograft transplantation for such patients . The role of immunosuppression is being evaluated. East Afr Med J, 1996 Jan, 73(1), 40 - 3 Cholera: its story in Africa with special reference to Kenya and other east African countries; Waiyaki PG; Cholera is an acute often fatal infection whose aetiology, epidemiology, clinical presentation, management, prevention and control are well understood . However, the disease has recently re-emerged as a serious microbial threat due to environmental pressures, increased international travel, population migrations and the emergence of new strains of "old" pathogens . This paper emphasizes, the significance of putting into place quick surveillance methods backed by laboratory services and the importance of creating a rapid response capacity and mechanics to outbreaks. East Afr Med J, 1996 Jan, 73(1), 32 - 4 Rabies: the emergence of a microbial threat; Borus P; Since 1912 when rabies was first confirmed in Kenya, the disease has largely existed in varying degrees of occurrence . Spatial and temporal distribution of cases of animal rabies are well documented . Records of human cases have not been as detailed as the veterinary cases . The principal animal reservoir for rabies has been the dog . Save for the 1960s, rabies outbreaks occurred in the country for most part of this century . The 1980's witnessed a dramatic upward swing in the number of cases reported annually . Over the years an enzootic pattern covering most parts of the country emerged . This article reviews available data showing rabies as an emerging microbial threat in Kenya. East Afr Med J, 1996 Jan, 73(1), 13 - 9 Human immunodeficiency virus infection and AIDS in east Africa: challenges and possibilities for prevention and control; Mhalu FS et al.; Human immunodeficiency infection and AIDS are a major recent microbial infection in east Africa with serious health and socioeconomic impacts in the region . At present HIV infection and AIDS account for more than 50% of adult medical admissions into some of the national and provincial hospitals as well as for 10-15% of paediatric admissions . AIDS is also at present the commonest cause of death among those aged 15-45 years . Tuberculosis, a closely associated disease to HIV infection, has increased more than three fold in some countries in the region . The prevalence of HIV infection currently ranges from 10-30% among adults in urban areas and from less than 1% to 25% in adults in rural areas; since this prevalence is still rising, the full impact of the AIDS problem in east Africa is yet to be realised . This is different from the situation in many developed countries where AIDS is no longer a priority health issue and where peak prevalences of the infection have been reached . The differences in HIV prevalences between east Africa and developed countries are due to poverty, ignorance, high prevalence of other STDs and associated cultural and traditional practices which prevail and facilitate HIV transmission in the region . While more than 80% of HIV infection in east Africa is transmitted through heterosexual intercourse, 5-15% of cases are perinatally transmitted and the remaining cases are transmitted through blood and blood products . While a lot of scientific advances have been made in immunopathology of AIDS, diagnostics and in social behavioural studies, we are still a long way towards getting curative therapy and or effective preventive vaccines . Recent discovery that use of zidovudine can significantly reduce perinatal HIV transmission is an additional breakthrough . While knowledge and tools for preventing HIV transmission are available in the world, prospects for AIDS control in east Africa appear gloomy unless major efforts are made in the reduction of poverty, ignorance and in the control of other common sexually transmitted diseasesPIP: HIV/AIDS poses a major health and socioeconomic burden in east Africa . More than 50% of adults and 10-15% of children admitted to some of the national and provincial hospitals in the region have HIV/AIDS . AIDS is the leading cause of death among people aged 15-45 . Tuberculosis, an opportunistic infection associated with HIV infection, has risen more than three times in some east African countries . 10-30% of adults in urban areas and from less than 1% to 25% of adults in rural areas are infected with HIV . In fact, HIV prevalence is still increasing; so the full impact of AIDS in eastern Africa has not yet occurred . Yet in many developed countries, HIV infection has peaked and HIV/AIDS is no longer a top priority . Poverty, ignorance, high prevalence of other sexually transmitted diseases (STDs), and associated cultural and traditional practices (e.g., inheritance of widows and polygamy), which abound and facilitate HIV transmission, account for the differences in HIV prevalence between east Africa and developed countries . Heterosexual intercourse accounts for more than 80% of HIV infections in east Africa . Perinatal transmission accounts for 5-15% of HIV cases . Blood and blood products transmitted HIV in the remaining cases . There have been considerable scientific advances in immunopathology of AIDS, diagnostics, and social behavioral studies; yet a cure and/or an effective vaccine is not in the near future . Recently, scientists have discovered that a mother's use of zidovudine during the last weeks of pregnancy and labor and zidovudine administered to the newborn significantly reduce perinatal HIV transmission . The knowledge and tools for preventing HIV transmission are available worldwide, but AIDS control in eastern Africa is apt to be for naught unless strong efforts are implemented toward the reduction of poverty, ignorance, and in the control of other common STDs . Curr Opin Periodontol, 1996, 3, 45 - 58 Early-onset periodontitis; Novak MJ et al.; The group of periodontal diseases known as the early-onset periodontal diseases are defined by the age of onset of periodontal destruction, distribution of lesions, association of disease with specific microbial infections, and identification of characteristic alterations, in the host response . Significant progress has recently been made in our understanding of the etiology of juvenile and rapidly progressive periodontitis . Considerable evidence points to a familial pattern of disease; both localized and generalized forms of disease may be observed in the same family . The exact mode of inheritance remains unclear, and disease may be the result of a complex interplay between genetically determined alterations of the host response and a specific bacterial challenge . Both neutrophil function and immunoglobulin response are altered and appear to be characteristic of an immunologic hyperresponsiveness . Bacterial colonization by Actinobacillus actinomycetemcomitans (serotype b) and/or Porphyromonas gingivalis appears to be the primary initiator of disease . Evidence suggests that lesion distribution may be a function of the nature of the infecting agent and the characteristics of the immune response. Arch Neurol, 1996 Jan, 53(1), 18 - 22 Emerging viral infections; Johnson RT; New microbial threats to human health and survival have arisen intermittently over the centuries . The increasing global population provides greater numbers of hosts for mutational evolution and sufficient hosts to ensure maintenance of new agents; the magnitude and modes of modern travel make a larger population of susceptible people accessible and provide rapid spread of infectious agents . This milieu is particularly hospitable for viruses; therefore, in the future we can anticipate increasing numbers of emerging viral diseases . This article on microbial threats has been limited to viruses associated with neurological diseases. Can J Microbiol, 1996 Jan, 42(1), 66 - 71 Halopicolinic acids, novel products arising through the degradation of chloro- and bromo-biphenyl by Sphingomonas paucimobilis BPSI-3; Davison AD et al.; Sphingomonas paucimobilis BPSI-3 was previously isolated from a mixed microbial consortium growing on biphenyl as the sole source of carbon and energy . Transformation of 4-chlorobiphenyl (4CBP) was demonstrated by this strain, although little or no growth was observed . In minimal salts medium supplemented with 4CBP or bromobiphenyl and dextrose, yellow coloured product(s) were rapidly formed . Gas chromatography-mass spectrometry (GC-MS) revealed single-ring N-heterocyclic compounds that were identified as halopicolinic acids . We believe this to be the first report of such compounds being formed via biological transformation of halobiphenyls . A mechanism is proposed for their formation. Berl Munch Tierarztl Wochenschr, 1996 Jan, 109(1), 28 - 31 {Effect of premortal stress on the endogenous microbial contamination of broiler carcasses}; Mengert U et al.; A study was undertaken to determine the interrelation between premortal stress and endogenic contamination . In 370 broilers the influence of transport, of a 48-h rest period and of an additional heat stress on the bacterial translocation was investigated . Even the normal transportation caused a contamination rate of nearly 50 per cent . 10 per cent of the transported animals suffered from a bacteriaemia induced by stress . The control group showed a significantly lower contamination rate . The rest time reduced the frequency of contamination . Heat stress did not have an influence on the contamination rate . The conclusion is that stress is aggravating the microbial risk for the consumer of poultry meat . Further research is necessary for characterizing the process of endogenic contamination. Arch Insect Biochem Physiol, 1996, 31(2), 119 - 33 Immune response in insects: the role of phenoloxidase in defense reactions in relation to melanization and sclerotization; Marmaras VJ et al.; It is well known that activated prophenoloxidase (proPO) plays an important role in cuticular melanization and sclerotization . In addition, studies dealing with immune response of insects suggest that phenoloxidase (PO) is also critical in the defense reactions of insects against invaders . proPO is activated by elicitors derived from microbial cell wall components such as peptidoglycan, beta-1,3-glucan, and lipopolysaccharide (LPS) . According to our recent studies we proposed a model clarifying the role of PO in both cellular and humoral immune responses . LPS triggers Ceratitis capitata hemocytes via induced protein tyrosine phosphorylation to release biologically active molecules, including p47 and proPO-activators . Furthermore, hemocytes in response to LPS facilitate clearance of LPS from the hemocoel of medfly . The effector molecules involved in the LPS clearance are hemocyte surface-associated p47 (mp47), soluble p47 (sp47), activated proPO, and tyrosine . A similar LPS clearance system in the integument of medfly in vitro was also demonstrated . According to our data, the proposed mechanism for LPS clearance from hemocoel and from integument is the crosslinking of LPS to p47 or certain integumental proteins via the intermediacy of reactive tyrosine derivatives generated by PO activity, as is the case for cuticular protein-chitin crosslinks during sclerotization . We also demonstrated that metabolites of the eumelanin biosynthesis and not melanin itself or N-acetyldopamine (NADA), the key precursor of sclerotizing agent, were necessary for the immune responses by hemocytes and integument. Adv Biochem Eng Biotechnol, 1996, 53, 89 - 118 Production of rhamnolipid biosurfactants; Ochsner UA et al.; Biosurfactants are of increasing interest due to their broad range of potential applications . A large variety of microbial surfactants is known at present, some of which may be used for specific applications . Towards the large scale industrial production of biosurfactants, the physiology, biochemistry and genetics of biosurfactant synthesis has to be well understood . A fully integrated process has to be developed, allowing high productivities under optimized conditions . In the past few years, we have investigated the molecular biology of rhamnolipid biosynthesis have been partially purified and characterized . The structural and regulatory genes encoding the rhamnolipid synthesis pathway have been isolated and characterized . The knowledge of the complex mechanisms involved in rhamnolipid synthesis facilitates the overproduction of these extracellular compounds . Furthermore, the transfer of the relevant genes into other species allows the production of rhamnolipids in heterologous hosts under controlled conditions . An integrated process for the production of rhamnolipids on an industrial scale has been developed . This process involves continuous cultivation under optimized media and growth conditions and makes use of refined methods of cell recycling, gas exchange and downstream processing, thus allowing high yields and productivities. Arch Ophthalmol, 1996 Jan, 114(1), 84 - 7 Empirical or culture-guided therapy for microbial keratitis? A plea for data; McDonnell PJ; In the United States, about 30,000 bacterial corneal ulcers are treated annually . Compared with 100, or even 20, years ago ophthalmologists today have available to them many diagnostic tools (including special media and bacterial identification techniques), and in impressive assortment of antibiotics . Many reviews and book chapters describe the uses of microbiologic studies--from Gram and Giemsa staining to media inoculation to immunofluorescence and even molecular genetics--to identify causative organisms . This literature also describes the formulas for preparation of highly concentrated, "fortified" antibiotics for initial treatment of bacterial keratitis, until culture and sensitivity results are available to guide modifications in therapy . It would seem, therefore, that "experts" in the field of corneal and external diseases have reached consensus on an appropriate initial microbiologic evaluation and treatment of suspected microbial keratitis, and the large body of literature on this topic might be considered to represent practice guidelines . It comes as a surprise to many that these published "guidelines" apparently are routinely ignored in current clinical practice. Am J Med, 1995 Dec 29, 99(6A), 45S - 52S; discussion 52S-53S Do monoclonal antibodies and anticytokines still have a future in infectious diseases? Cohen J, Heumann D, Glauser MP. The continuing high mortality of septic shock has prompted a major effort by the research community to identify novel therapeutic targets . These targets can be conveniently grouped into (1) those derived from microbial components or products; (2) inflammatory mediators; and (3) effector molecules . Many of the experimental, so-called adjunctive agents developed have been monoclonal antibodies or anticytokine molecules of various kinds, and some have progressed into clinical trial . Unfortunately, these trials have failed to show unequivocal survival benefit for patients in shock, prompting a reappraisal of our approach to these agents . In this article, we discuss the possible reasons for these failures: (1) the targets are wrong; (2) the agents are inappropriate; or (3) the trial design is flawed . It would be premature to conclude that adjunctive agents have no future in the therapy of sepsis, but identifying the correct agent, and perhaps more importantly, the correct target population, is going to be more difficult than was at first believed. J Immunol, 1995 Dec 15, 155(12), 5700 - 4 An ospA frame shift, identified from DNA in Lyme arthritis synovial fluid, results in an outer surface protein A that does not bind protective antibodies; Fikrig E et al.; Passive immunization with murine or human Abs to outer surface protein A (OspA) can protect mice against Borrelia burgdorferi, but OspA Abs elicited during natural infection in mice or humans are unable to clear the spirochete from the infected host . To examine Ab binding by OspA during the course of human infection, we amplified the operon encoding full-length ospA and ospB from synovial fluids of a patient with chronic Lyme arthritis, the first such recoveries from human material, at four separate time points over 4.5 mo, and expressed OspA in Escherichia coli . OspA mAbs that passively protected mice from infection did not bind one of the expressed OspAs, because of a deletion in ospA that resulted in a frame shift and premature stop codon near the carboxyl terminus . However, expressed OspA from a later synovial fluid sample did not contain this deletion . Thus, although altered forms of OspA, which potentially can influence host immune effectiveness, do occur in the human host, they cannot be the only factors responsible for microbial persistence. Arch Latinoam Nutr, 1995 Dec, 45(4), 317 - 21 {Proteolytic activity control in fish silage}; Tome E et al.; Microbial fish silage was produced from ground low commercial fish species added with 15% mollase and stored at 35 degrees C . Experiment were conducted to evaluate the effect of the addition of a commercial preparation of bromeline (0.20-0.90%) to the fish silage . Liquefaction was evaluated measuring both the product consistency and its non protein nitrogen content . Results indicate that bromeline addition (0.7 or higher), increases dramatically the rate of proteolysis and decreases the time required for liquefaction from 15 days (no enzyme control) to only 12 hours . Therefore, the use of the commercial preparation of bromeline is highly recommended for silage production. J Small Anim Pract, 1995 Dec, 36(12), 542 - 6 Periodontal disease and leucopenia; Lonsdale T; Periodontal disease is rare in nature but widespread in domestic dog and cat populations . Unnatural diets are known to facilitate the buildup of oral microbial communities which then interact with host-immune defences giving rise to periodontitis . Eight of 14 animals undergoing dental treatment and dietary change at a suburban veterinary practice were investigated and found to have low leucocyte counts . Follow-up testing revealed changes averaging a 77.7 per cent increase with concomitant 'subjective good health' . These findings serve to cast doubt on the commonly used haematological reference ranges where the subject animals may have suffered from periodontal disease . The demonstrated reversibility of white cell depression associated with periodontal disease should provide a focus for further research. Fundam Appl Toxicol, 1995 Dec, 28(2), 274 - 82 Mucous cell metaplasia in the airways of rats exposed to machining fluids; Gordon T et al.; Occupational exposure to microbial-contaminated machining fluids is associated with a variety of adverse pulmonary effects including chronic bronchitis and increased sputum production . We have previously demonstrated in F344 rats that inhaled endotoxin can increase the amount of stored intraepithelial mucosubstances (Vs) in the respiratory tract . The purpose of the present study was to examine the effect of endotoxin-contaminated machining fluid aerosols on mucous production . Rats were exposed to aerosols of pyrogen-free water, 1 or 10 mg/m3 used machining fluid, or 10 mg/m3 unused machining fluid for 3 hr/day for 3 days . Twenty-four hours after the final exposure, right lung lobes were lavaged and the nasal cavity and left lung were fixed in formalin . The amount of Alcian blue/periodic acid-Schiff-stained mucosubstances was determined by morphometry . Exposure to 10 mg/m3 used machining fluid (equivalent to 0.8 micrograms/m3 endotoxin) produced a significant increase in Vs in the epithelial lining of both the nasal septum and intrapulmonary airways . These changes in Vs were accompanied by a significant increase in total cells and neutrophils in the lavage fluid . No changes in stored mucosubstances or lavage parameters were found in animals exposed to 1 mg/m3 used machining fluid aerosols . A significant increase in Vs was observed in the nasal septum but not in the intrapulmonary airways of animals exposed to 10 mg/m3 unused machining fluids (no measurable endotoxin) . These results suggest that in addition to endotoxin, nonendotoxin components of machining fluids may contribute to the increase in sputum and chronic bronchitis reported for workers exposed to machining fluid aerosols. Fundam Appl Toxicol, 1995 Dec, 28(2), 155 - 66 Water chlorination: essential process or cancer hazard? Bull RJ, Birnbaum LS, Cantor KP, Rose JB, Butterworth BE, Pegram R, Tuomisto J. Chlorine has been successfully used for the control of waterborne infectious disease for nearly a century . In the 1970s it was found that chlorine reacted with natural organic matter present in surface waters to produce disinfection by-products (DBP) . Concern focused initially on the trihalomethanes (THM), but a wide variety of DBPs are now known to result from chlorination . Chlorination of drinking water has been one of the most effective public health measures ever undertaken . There are a number of alternatives to chlorination that are in active use in many parts of the world, but the risks associated with their by-products are even less well established than for chlorination . Moreover, the use of these alternatives vary in their effectiveness and some require greater sophistication in their application . This can mean less protection to public health as a result of inappropriate application and control . Therefore, hazards associated with the use of such a clearly beneficial process as chlorination must be carefully considered not only in an absolute sense, but also in the context of alternative approaches for producing a safe drinking water . The key question is whether the hazards associated with by-products have been sufficiently well established to warrant regulations that will undoubtedly have both positive and negative impacts on the public health . This symposium examined the toxicological and epidemiological data on chemical hazards associated with chlorination and attempted to measure this hazard against competing microbial risks . The first presentation discussed the available analytical epidemiological studies . A second presentation dealt with the importance of chlorination to the prevention of waterborne infectious disease . Pharmacokinetic, mechanistic, and modeling information on the prototypical DBP, chloroform, were discussed and contrasted with data on brominated THMs to determine if it was scientifically appropriate to regulate THMs as a single toxicological class . The fifth presentation dealt with the carcinogenic properties of a potent mutagen that is produced by chlorination . The final presentation discussed the haloacetates, carcinogenic DBPs whose concentrations approach and occasionally exceed those of the THMs . Clearly, there is a need to carefully weigh these different types and sometimes competing risks when considering the delivery of drinking water to ever-increasing populations for which there are finite sources of fresh water. Scand J Work Environ Health, 1995 Dec, 21(6), 450 - 9 Cancer risk among workers in biomedical research; Cordier S et al.; OBJECTIVE: This epidemiologic study was undertaken after a cluster of five cases of rare forms of cancer (bone sarcoma, non-Hodgkin's lymphoma) had been observed among biomedical research workers at the Pasteur Institute in Paris to ascertain whether their disease was connected with exposure during this research . METHODS: A mortality study included 3765 people who worked at the Pasteur Institute between 1971 and 1986 and were followed until the end of 1987 . Within this cohort a nested case-referent study included 23 cases of cancer {non-Hodgkin's lymphoma (6), multiple myeloma (1), leukemia (3), pancreatic cancer (7), bone cancer (3), brain tumor (3)}, and four referents per case, matched for gender and year of birth . RESULTS: Total mortality from cancer was less than expected, the standardized mortality ratio (SMR) being 72 for the men and 82 for the women . Among the women the proportion of pancreatic cancer cases was larger than expected {SMR 490, 95% confidence interval (95% CI) 158-1144}, as was the number of brain cancer cases (SMR 239, 95% CI 48-696) . Among the men, mortality from bone cancer was greater than expected (SMR 553, 95% CI 62-2006) . In the nested case-referent study, more cases than referents had worked in the areas of molecular biology {odds ratio (OR) 7.1, 95% CI 1.5-33} and microbial genetics (OR 6.7, 95% CI 1.3-35) . These cases especially included non-Hodgkin's lymphoma and bone cancer . Associated with this finding was the fact that more cases had used certain chemicals, including ethidium bromide, acrylamide, methylnitronitrosoguanidine and ethylmethanesulfonate, and radioactive compounds (essentially 32phosphorus) . CONCLUSIONS: As the products used are potent genotoxicants, the present findings suggest that work in biomedical research might well involve an increased risk of certain types of cancer; this conclusion should be balanced by the fact that two of the five index cases were included in the mortality study and four in the nested case-referent study. J Ind Microbiol, 1995 Dec, 15(6), 457 - 71 Review: biocatalytic transformations of ferulic acid: an abundant aromatic natural product; Rosazza JP et al.; In this review we examine the fascinating array of microbial and enzymatic transformations of ferulic acid . Ferulic acid is an extremely abundant, preformed phenolic aromatic chemical found widely in nature . Ferulic acid is viewed as a commodity scale, renewable chemical feedstock for biocatalytic conversion to other useful aromatic chemicals . Most attention is focused on bioconversions of ferulic acid itself . Topics covered include cinnamoyl side-chain cleavage; nonoxidative decarboxylation; mechanistic details of styrene formation; purification and characterization of ferulic acid decarboxylase; conversion of ferulic acid to vanillin; O-demethylation; and reduction reactions . Biotransformations of vinylguaiacol are discussed, and selected biotransformations of vanillic acid including oxidative and nonoxidative decarboxylation are surveyed . Finally, enzymatic oxidative dimerization and polymerization reactions are reviewed. Chem Biol, 1995 Dec, 2(12), 775 - 80 Cleaning up our own backyard: developing new catabolic pathways to degrade pollutants; Minshull J; Microbial-based strategies for pollution control require metabolic pathways by which man-made compounds may be degraded . Recombination-based mutagenesis and selection procedures may be able to mimic the evolution of catabolic pathways and generate enzymes with novel specificities. J Accid Emerg Med, 1995 Dec, 12(4), 286 - 7 Do patients presenting to accident and emergency departments with the sensation of a foreign body in the eye (gritty eye) have significant ocular disease? Jayamanne DG. A total of 1155 consecutive patients selected on symptomatology alone who presented to an ophthalmic accident and emergency (A&E) department over a period of 3 months from a total patient population of 3522 were assessed . The study showed that about 96% of the patients reporting a sensation of a foreign body in their eye had no serious ocular disease . Most patients had a superficial mechanical injury or minor microbial infection . Steroid drugs were prescribed to only 2.1% of the patients . Topical steroid treatment was contraindicated in 0.35% of the patients . A low rate of morbidity was noted, suggesting that these patients could have been treated adequately in a general A&E department rather than an ophthalmic department. Gastroenterologist, 1995 Dec, 3(4), 329 - 44 Vitamin B12 in health and disease: part I--inherited disorders of function, absorption, and transport; Kapadia CR; All of vitamin B12 in nature is of microbial origin . Cobalamin, as vitamin B12 should correctly be termed, is a large polar molecule that must be bound to specialized transport proteins to gain entry into cells . Entry from the lumen of the intestine under physiological conditions occurs only in the ileum and only when bound to intrinsic factor . It is transported into all other cells only when bound to another transport protein, transcobalamin II . Congenital absence or defective synthesis of intrinsic factor or transcobalamin II result in megaloblastic anemia . The Immerslund-Graesbeck syndrome, a congenital defect in the transcellular transport of cobalamin through the ileal cell during absorption, also presents with megaloblastic anemia, but with accompanying albuminuria . In most bacteria and in all mammals, cobalamin regulates DNA synthesis indirectly through its effect on a step in folate metabolism, the conversion of N5-methyltetrahydrofolate to tetrahydrofolate, which in turn is linked to the conversion of homocysteine to methionine . This reaction occurs in the cytoplasm, and it is catalyzed by methionine synthase, which requires methyl cobalamin (MeCbl), one of the two coenzyme forms of the vitamin, as a cofactor . Defects in the generation of MeCbl (cobalamin E and G diseases) result in homocystinuria; affected infants present with megaloblastic anemia, retardation, and neurological and ocular defects . 5'-Deoxyadenosyl cobalamin (AdoCbl), the other coenzyme form of cobalamin, is present within mitochondria, and it is an essential cofactor for the enzyme Methylmalonyl-CoA mutase, which converts L-methylmalonyl CoA to succinyl CoA . This reaction is in the pathway for the metabolism of odd chain fatty acids via propionic acid, as well as that of the amino acids isoleucine, methionine, threonine, and valine . Impaired synthesis of AdoCbl (cobalamin A or B disease) results in infants with methylmalonic aciduria who are mentally retarded, hypotonic, and who present with metabolic acidosis, hypoglycemia, ketonemia, hyperglycinemia, and hyperammonemia . Megaloblastic anemia does not develop in these children because adequate amounts of MeCbl are present, but the effect of methylmalonic acid on marrow stem cells may give rise to pancytopenia . Congenital absence of reductases in the cytoplasm, which normally reduce the cobalt atom in cobalamin from its oxidized to its reduced state (cobalamin C and D diseases), results in impaired synthesis of both MeCbl and AdoCbl . Both methylmalonic aciduria and homocystinuria therefore develop in these children, and they present with megaloblastosis, mental retardation, a host of neurological and ocular disorders, and failure to thrive; however, they do not have hyperglycinemia or hyperammonemia . A similar biochemical profile and clinical presentation is also seen in cobalamin F disease, which results from a defect in the release of cobalamin from lysosomes, following receptor-mediated endocytosis of the transcobalamin II-cobalamin complex into cells . It is important to recognize these inborn errors of cobalamin absorption, transport, or function as soon after birth as possible, because most respond (in some patients more fully than others) to parenteral administration of cobalamin . Delays in diagnosis can lead to grave clinical consequences. Cardiovasc Surg, 1995 Dec, 3(6), 711 - 4 Contained rupture of infected abdominal aortic aneurysm due to systemic candidiasis; Ikeda M et al.; A case is reported of contained rupture of an infected abdominal aortic aneurysm as a result of vascular catheter-associated candidiasis . Candida was identified in the mural thrombus taken from the aneurysm during surgery . Since Candida has played an important role in nosocomial infection and catheter-associated sepsis during the past decade, it is becoming an increasingly significant pathogen in microbial arteritis . Successful treatment included axillobifemoral bypass grafting followed by a complete excision of the infected aneurysm, retroperitoneal drainage and long-term antifungal therapy . Although infected abdominal aortic aneurysm is not common, without surgical intervention it may lead to serious consequences, such as rupture or uncontrollable sepsis . As the clinical symptoms of this disease are minimal and non-specific during the early stages, a high index of suspicion for this condition is essential for precise diagnosis and successful treatment . Moreover, initial treatment of candidiasis is especially important to prevent this rare but fatal late complication, infected abdominal aortic aneurysm. Int J Gynaecol Obstet, 1995 Dec, 51(3), 211 - 8 Intrauterine death and infections during pregnancy; Moyo SR et al.; OBJECTIVE: To elucidate whether microbial infections are involved in the etiology of intrauterine death . METHODS: One hundred four cases of stillbirth of unknown etiology and 96 age- and parity-matched referents with live births were analyzed with respect to microbial infection by cultures from the placenta, endocervix and internal organs of the fetuses, external sites of the babies and fetuses, and by serology for bacteria, viruses and Toxoplasma gondii . RESULTS: In 17 cases in whom no other infectious agent was diagnosed, Escherichia coli was isolated from the placenta and one or more internal fetal organs . Tests for Treponema pallidum and Toxoplasma gondii were more frequently positive in cases than in referents (O.R . 8.3 and 3.9, respectively) . There was no increased risk for intrauterine death in women with human immunodeficiency virus, cytomegalovirus, herpes simplex virus or rubella virus . CONCLUSIONS: Our findings indicate that infections remain an important cause of intrauterine death in Zimbabwe. J Clin Invest, 1995 Dec, 96(6), 2578 - 82 Endogenous production of interleukin 15 by activated human monocytes is critical for optimal production of interferon-gamma by natural killer cells in vitro; Carson WE et al.; Natural killer (NK) cells are large granular lymphocytes that constitutively express functional IL-2 receptors . We have shown that recombinant human IL-15 uses the IL-2 receptor to activate human NK cells and can synergize with recombinant human IL-12 to stimulate NK cell production of IFN-gamma in vitro . IFN-gamma production by NK cells is critical in the prevention of overwhelming infection by obligate intracellular microbial pathogens in several experimental animal models . Herein, we demonstrate that human monocytes produce IL-15 protein within 5 h of activation with LPS . Using an IL-15-neutralizing antiserum in a coculture of LPS-activated monocytes and NK cells, we demonstrate that monocyte-derived IL-15 is critical for optimal NK cell production of IFN-gamma . Endogenous IL-15 activates NK cells through the IL-2 receptor, and with endogenous IL-12, regulates NK cell IFN-gamma after monocyte activation by LPS . These in vitro studies are the first to characterize a function for endogenous IL-15, and as such, suggest an important role for IL-15 during the innate immune response . IL-15 may be an important ligand for the NK cell IL-2 receptor in vivo. Rev Sci Tech, 1995 Dec, 14(4), 913 - 23 Limitations, definitions, principles and methods of risk analysis; North DW; Decisions on veterinary biologicals involve large uncertainties, complexities which cut across many scientific and technical disciplines, and large potential adverse impacts on public health and on important sectors of the economy . How should risk assessment help to guide the decision process on veterinary biologicals? How can risk assessment practices be harmonized internationally, given the different regulatory traditions and institutions of different countries? A broad view of risk assessment is needed, that risk assessment is a framework for summarizing applicable scientific judgement in support of regulatory decision-making . Support for this view of risk assessment is found in the major reports which have defined risk assessment as currently practised by many regulatory agencies in the United States of America (USA) . However, some interested and affected parties perceive risk assessment in the USA as overly quantitative and narrowly focused on regulatory standards for carcinogens . An example of risk assessment for microbial contamination indicates how quantitative methods can be used when data are sparse and decisions must be made in the face of great uncertainty . Such quantitative methods can be used to improve communication about risk, to promote consensus in support of controversial decisions, and to identify valuable opportunities for research to reduce the important sources of uncertainty. Oral Microbiol Immunol, 1995 Dec, 10(6), 365 - 71 Viability of four putative periodontal pathogens and enteric rods in the anaerobic transport medium VMGA III; Ali RW et al.; The aim of this study was to determine the survival in VMGA III of Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, Actinobacillus actinomycetemcomitans and enteric rods in laboratory cultures as well as in subgingival plaque samples . Laboratory strains of the 4 putative periodontal pathogens and Escherichia coli were used in the laboratory part of this study . Also, 31 subgingival plaque samples were obtained from 22 periodontal patients and stored in VMGA III . Each sample, from both the laboratory and the clinical parts, was divided into 3 portions . One portion was cultured within a few hours of collection (baseline), while the second was processed after 24 h (day 2) and the third 48 h later (day 3) . The results of the clinical part indicate that the detection frequencies of all 4 periodontal pathogens and their levels in positive samples decreased, to different degrees, by day 2 and decreased further by day 3 . Enteric rods were not detected in base line samples . However, they were present in 16.1% and 22.6% of day 2 and day 3 samples, respectively . Similarly, the laboratory results demonstrate a significant decrease in the levels of the 4 periodontal pathogens tested by day 2 and day 3, whereas the opposite occurred for E . coli . P . gingivalis, P . intermedia, and F . nucleatum survived better in the presence of E . coli than alone, whereas A . actinomycetemcomitans survived less well when co-inoculated with E . coli . VMGA III appears to maintain microbial population ratios for periods up to 24 h . After 24 h, the multiplication of enteric organisms may alter the original proportions of the sample. Biophys J, 1995 Dec, 69(6), 2226 - 33 Method for immobilizing microbial cells on gel surface for dynamic AFM studies; Gad M et al.; The processes of cell growth and budding of the yeast cells Saccharomyces cerevisiae, which were gently immobilized on 3% agar and submerged in culture medium, were successfully imaged with an atomic force microscope for 6-7 h . Similar experiments on chemically fixed cells did not detect any appreciable change in their appearance except in a few scannings at the very beginning, indicating that the dissolution of agar and/or scraping of its surface by the scanning tip, if any, did not significantly interfere with the images taken thereafter . The increment in the height of many of the untreated cells, accompanied by their lateral enlargement, was taken as an indication of successful imaging of the growth process of yeast cells, together with an image of a growing daughter cell attached to its mother cell. Appl Microbiol Biotechnol, 1995 Dec, 44(3-4), 393 - 8 Purification and characterization of a newly screened microbial peptide amidase; Stelkes-Ritter U et al.; A microbial peptide amidase was found in a limited screening and purified about 500-fold from Stenotrophomonas maltophilia . The native enzyme has a molecular mass of 38 kDa (gel filtration) . The sequence of the first 16 amino acids was determined by Edman degradation . The isoelectric point was found to be around 5.8 . The peptide amidase exhibited a pH optimum of 6.0 and a temperature optimum of about 39-45 degrees C . The enzyme is stable in 50 mM TRIS/HCl, pH 7.5, at 30 degrees C, and the residual activity was found to be above 90% after 1 week of incubation . The biocatalyst is not inhibited by potential inhibitors like Hg2+, EDTA, D-cycloserine or dithiothreitol and only weakly influenced by inhibitors of serine proteases . The peptide amidase deamidates selectively C-terminal amide groups in peptide amides without hydrolysing internal peptide bonds or amide functions in the side-chain of glutamine or asparagine . Unprotected amino acid amides are not hydrolysed . The enzyme is stereoselective with regard to L-enantiomers in the C-terminal position. Toxicol Lett, 1995 Dec, 82-83, 159 - 65 Modulators of signal transduction as cancer chemotherapeutic agents--novel mechanisms and toxicities; Gescher A; The recent explosion of knowledge of the molecular basis of regulation of growth and differentiation has engendered the search for novel anticancer drugs which act via interference with cellular signal transduction pathways . Among the agents which are under investigation are modulators of protein kinases, for example quercetin and bryostatin 1 . Both compounds have undergone phase 1 clinical evaluation . The major dose-limiting toxicities of quercetin and bryostatin are nephrotoxicity and myalgia, respectively . Novel analogs of the microbial product staurosporine, a potent kinase inhibitor, possess intriguing specificities for different kinases. Trends Biotechnol, 1995 Dec, 13(12), 527 - 37 Engineering liposomes for drug delivery: progress and problems; Gregoriadis G; Liposomes were first proposed and tested as a drug delivery system 25 years ago . Since then, advances in our understanding of the fate and behaviour of liposomes at the cellular and subcellular level in vivo have allowed the rational design of constructs for use in the treatment and prevention of disease, both in experimental animals and clinically . The involvement of several liposome-based biotechnology companies in the early 1980s, paralleled by great leaps in liposome technology, has culminated in the design and licencing of formulations for the treatment of certain microbial infections and cancers, and the first liposome-based vaccine for use in humans. Allerg Immunol (Paris), 1995 Dec, 27(10), 375 - 6 {Candida vasculitis}; Bourrain JL et al.; Apart from direct skin attacks, yeasts of the Candida group hold a place that is much discussed in the occurrence of immuno-allergic reactions of the skin, because of their usual saprophytic characteristics . We report here on observation of a patient of 62 years who presented with a secondary purpuric maculo-papular eruption making a tri-symptom of Gougerot, for which we have incriminated the role of the immunogenicity of Candida . Skin histology confirmed the presence of an angiitis and necrotising capillarity . An intradermal test with candidine reproduced the skin symptoms and histology, whilst other microbial and fungal tests remained within normal limits . Other examinations showed the transitory presence of circulating immune complexes, although the remainder of the immunity factors were negative . A digestive focus of Candida was present . Recovery occurred after systemic mycological treatment was started, though there was a return of symptoms after a temporary initial check. Hautarzt, 1995 Dec, 46(12), 858 - 9 {Pruritus of the artificial vagina of a transsexual patient caused by gonococcal infection}; Haustein UF; Report on a 31-year-old male-to-female transsexual prostitute with a 3-week history of worsening pruritus and dysaesthesia in her neovagina, which had been constructed by means of penile skin invagination . The microbial diagnostic tests for sexually transmitted diseases (STD) showed gonococcal infection in the high vagina, while none was found in the urethra . Other STD and allergic contact sensitization were excluded . Despite the regularly stratified epithelium of the neovagina, which is physiologically resistant to gonococcal infection, this possibility must be considered in transsexual prostitutes. Dig Dis Sci, 1995 Dec, 40(12), 2532 - 9 Morphometric analysis of intestinal mucins under different dietary conditions and gut flora in rats; Sharma R et al.; Elucidation of the mechanisms that alter the biosynthesis, turnover, and degradation of intestinal mucins is relevant to the understanding of both the normal gut ecosystem and various intestinal diseases . In this study image analysis was used to quantify the effects of diet and microbial flora on the mucin composition of goblet and deep crypt cells, the number and volume density of mucin-containing cells, and the staining density of their stored mucins in the small and large intestine of germ-free and conventionally maintained rats fed two different diets . One was a coarsely ground commercial rodent diet containing crude fiber of cereal origin and the other a purified diet composed of finely powdered ingredients, including cellulose as a source of fiber . The changes in mucin production were also analyzed in germ-free rats colonized with a human flora . Feeding a commercial diet reduced the volume density of cells containing neutral and sulfomucins in the jejunum of conventional rats and the staining density of neutral and acidic mucins in the germ-free rats . Both rat and human floras reduced the number of cells containing acidic and sulfomucins and the staining density of neutral mucins in the small intestine of animals fed on a purified diet . However, inoculation of human flora increased the staining density of stored neutral and sulfated mucins in the cells of the large intestine . The results demonstrate that the dietary changes are influential in modifying the amount and proportion of mucins in the small intestine and the microbial flora in the large intestine. Appl Environ Microbiol, 1995 Dec, 61(12), 4269 - 73 Rapid differentiation of bacterial species with multiple probes of different lengths in a single slot blot hybridization; Wang GC et al.; We describe a highly efficient method for dot and slot blot hybridizations with multiple oligonucleotide probes for high throughput identification of organisms and studies of microbial community structures . Several probes with distinct specificities were designed to have the same melting temperature but unique lengths by adding different numbers of nonspecific nucleotides to one end . All of the probes were mixed, labelled with 32P, and hybridized to one piece of membrane on which genes coding for 16S rRNAs from different bacterial species had been immobilized . After hybridization, the bound probes were eluted and resolved on a denaturing polyacrylamide gel and the identities of the genes coding for 16S rRNAs were read from an autoradiograph of the gel . The results from the application of this technique to pure actinomycete cultures are reported here. J Bacteriol, 1995 Dec, 177(24), 7186 - 93 Ferrichrome transport in Escherichia coli K-12: altered substrate specificity of mutated periplasmic FhuD and interaction of FhuD with the integral membrane protein FhuB; Rohrbach MR et al.; FhuD is the periplasmic binding protein of the ferric hydroxamate transport system of Escherichia coli . FhuD was isolated and purified as a His-tag-labeled derivative on a Ni-chelate resin . The dissociation constants for ferric hydroxamates were estimated from the concentration-dependent decrease in the intrinsic fluorescence intensity of His-tag-FhuD and were found to be 0.4 microM for ferric aerobactin, 1.0 microM for ferrichrome, 0.3 microM for ferric coprogen, and 5.4 microM for the antibiotic albomycin . Ferrichrome A, ferrioxamine B, and ferrioxamine E, which are poorly taken up via the Fhu system, displayed dissociation constants of 79, 36, and 42 microM, respectively . These are the first estimated dissociation constants reported for a binding protein of a microbial iron transport system . Mutants impaired in the interaction of ferric hydroxamates with FhuD were isolated . One mutated FhuD, with a W-to-L mutation at position 68 {FhuD(W68L)}, differed from wild-type FhuD in transport activity in that ferric coprogen supported promotion of growth of the mutant on iron-limited medium, while ferrichrome was nearly inactive . The dissociation constants of ferric hydroxamates were higher for FhuD(W68L) than for wild-type FhuD and lower for ferric coprogen (2.2 microM) than for ferrichrome (156 microM) . Another mutated FhuD, FhuD(A150S, P175L), showed a weak response to ferrichrome and albomycin and exhibited dissociation constants two- to threefold higher than that of wild-type FhuD . Interaction of FhuD with the cytoplasmic membrane transport protein FhuB was studied by determining protection of FhuB degradation by trypsin and proteinase K and by cross-linking experiments . His-tag-FhuD and His-tag-FhuD loaded with aerobactin specifically prevented degradation of FhuB and were cross-linked to FhuB . FhuD loaded with substrate and also FhuD free of substrate were able to interact with FhuB. J Leukoc Biol, 1995 Dec, 58(6), 643 - 9 On the expression of nitric oxide synthase by human macrophages . Why no NO? Albina JE. The production of nitric oxide (NO) and its role in the anti-tumor and anti-microbial effects of rodent macrophages appears well established . In contrast, the circumstances required for its release from human monocytes/macrophages and its potential role in human pathology remain controversial . Evidence to be discussed suggests that NO is a redundant, autotoxic, immunosuppressive, and inefficient mediator of macrophage function . For these reasons, the expression of nitric oxide synthase as a rapid-response, high-output effector pathway may have been evolved out of the human monocyte/macrophage response repertoire or severely restricted in its expression . Hypothetical roles for a modest and circumscribed production of NO by human macrophages are proposed. Am J Pathol, 1995 Dec, 147(6), 1525 - 33 Pathology and emerging infections--quo vadimus? Schwartz DA, Bryan RT, Hughes JM. There have been dramatic changes in the occurrence of infectious diseases throughout the world in the previous two decades . The emergence of new microbial agents, and the reemergence of infections previously believed to be controlled, threatens the health of all populations . The emergence of these infectious diseases has occurred during a period of breakdown in the capabilities of the public health surveillance systems, prevention programs, and disease control efforts . Expertise in pathology is critical to provide a strong national control program for emerging and reemerging infectious diseases . Despite the many significant achievements made by pathologists in improving understanding of the pathogenesis and diagnosis of infectious diseases, the field of pathology remains largely oriented toward neoplastic diseases, and has not yet identified infectious disease diagnosis as an important component of anatomic pathology training and research, even in the face of the current threats posed by microbial agents . In addition, there is currently a dearth of infectious disease pathologists in the United States and elsewhere in the world, and the use of the autopsy, a prime pathological tool for diagnosis of emerging infections, is on the wane . The most serious problem is that no formal training program for infectious disease pathology currently exists in the United States or elsewhere in the world . As a consequence of this lack of training opportunities, there is a severe deficiency of young, well-educated pathologists with infectious disease expertise . This article explores the historical linkages between the disciplines of infectious diseases and pathology, and suggests that infectious disease pathology as a subspecialty be strengthened and training programs be initiated. Ugeskr Laeger, 1995 Nov 20, 157(47), 6585 - 6 {Landfill gas}; Laursen E et al.; In a landfill mixture of gases, consisting principally of methane and carbon dioxide, may be produced by microbial degradation of organic waste under anaerobic conditions . Methane is explosive at concentrations between 5 and 15% by volume . Other gases, for instance hydrogensulphide, mercury and ethane, may be emitted at low concentrations, but usually do not represent a health hazard following normal atmospheric dilution . Indoor climate may be affected, though, in cases of accumulation in closed spaces . A case is presented where two persons died following an explosion caused by lighting a cigarette in their house which was surrounded on three sides by a landfill . The explosion occurred after heavy precipitation on a day with low atmospheric pressure . Methane measurements showed values consistent with risk of explosion. Rev Esp Salud Publica, 1995 Nov-Dec, 69(6), 445 - 61 {Microbial resistance: what is to be done? Information from the panel of experts}; Mochales FB et al.; This document represents the recommendations of a panel of Spanish experts on antibiotic use and resistance . In a Task Force, under the auspices of the Spanish Ministry of Health and Consumer Affairs that took place in 1994 in Madrid, the members were gravely concerned about the national increase in antibiotic resistance . They analysed the development, evolution and spread of antibiotic resistance among community-acquired human bacterial pathogens in Spain, its relation with antibiotic consumption, and they proposed future surveillance strategies for monitoring the patterns of antibiotic use and consumption . Success will require a collective action among the producers (pharmaceutical industry), prescribers (doctors, veterinarians), dispensers (pharmacists), and consumers (patients) . Two similar documents have been recently published by the American Society of Microbiology and the World Health Organization showing the global concern about this topic. Crit Rev Food Sci Nutr, 1995 Nov, 35(6), 509 - 24 Use of edible coatings to preserve quality of lightly (and slightly) processed products; Baldwin EA et al.; Lightly processed agricultural products present a special problem to the food industry and to scientists involved in postharvest and food technology research . Light or minimal processing includes cutting, slicing, coring, peeling, trimming, or sectioning of agricultural produce . These products have an active metabolism that can result in deteriorative changes, such as increased respiration and ethylene production . If not controlled, these changes can lead to rapid senescence and general deterioration of the product . In addition, the surface water activity of cut fruits and vegetables is generally quite high, inviting microbial attack, which further reduces product stability . Methods for control of these changes are numerous and can include the use of edible coatings . Also mentioned in this review are coating of nut products, and dried, dehydrated, and freeze-dried fruits . Technically, these are not considered to be minimally processed, but many of the problems and benefits of coating these products are similar to coating lightly processed products . Generally, the potential benefits of edible coatings for processed or lightly processed produce is to stabilize the product and thereby extend product shelf life . More specifically, coatings have the potential to reduce moisture loss, restrict oxygen entrance, lower respiration, retard ethylene production, seal in flavor volatiles, and carry additives that retard discoloration and microbial growth. Med Hypotheses, 1995 Nov, 45(5), 421 - 6 Views on the sustenance of resistant Mycobacterium tuberculosis in the environment; Basu A et al.; Microbial resistance to conventional as well as newly introduced drugs is a hallmark feature of several infectious diseases, notably tuberculosis . It is hypothesized that the greater the selective pressure exerted by increasingly potent drugs, the more rapidly is an organism able to adapt to a drug-containing environment . The roles of drug-containing environments, and the immunological status of the host and bacterial molecular mechanisms of development of drug resistance to Mycobacterium tuberculosis have been examined and examples cited for implementation of modified drug regimens in tuberculosis-control programmes . The views expressed, albeit restricted to Mycobacterium tuberculosis, encourage consideration of drug regimens on a disease evolution basis as well as understanding of the natural rules that govern development and sustenance of drug resistance in the microbial world. Mycoses, 1995 Nov-Dec, 38(11-12), 489 - 94 Cell-surface hydrophobicity and lipolysis as essential factors in human tinea nigra; Gottlich E et al.; Hydrophobic adhesion of cells of the black yeast Hortaea werneckii (Horta) Nishimura & Miyaji, causative agent of human tinea nigra, was established by microbial adhesion to hexadecane (MATH) and adhesion to polystyrene, and compared with adhesion of other species of black yeasts . Additional ecophysiological tests were performed . Hortaea werneckii cells proved to have a high degree of hydrophobicity (98.5% MATH) . The species is unable to degrade keratin, but shows significant lipolytic activity . It is concluded that H . werneckii is a commensal, that shows lipophilic adhesion to human skin and survives by the assimilation of excretion products. J AOAC Int, 1995 Nov-Dec, 78(6), 1525 - 9 Application of triphenyltetrazolium chloride in microbial limit test of pharmaceuticals and cosmetics; Ohara MT et al.; An alternative method was developed for counting viable microbial contaminants in pharmaceuticals and cosmetics with insoluble materials and low bioburden . The method uses triphenyltetrazolium chloride (TTC) . The inhibitory effect of TTC on microbial growth observed by others was eliminated by adding it as an overlay, after incubation of sample in culture medium for 48 h . Six samples of pharmaceutical suspensions and 6 samples of cosmetics were evaluated by most-probable number (MPN) and pour-plate techniques with TTC in comparison with MPN with subculturing . The equivalence between MPN technique with TTC and subculturing was demonstrated for all samples and between pour-plate and subculturing for 10 samples . The differences were probably due to some error inherent in both techniques and not from application of TTC . Comparison of coefficients of variation showed that plate countings were more precise than the MPN method, as expected. Mol Plant Microbe Interact, 1995 Nov-Dec, 8(6), 960 - 70 Purification and characterization of beta 2-tomatinase, an enzyme involved in the degradation of alpha-tomatine and isolation of the gene encoding beta 2-tomatinase from Septoria lycopersici; Sandrock RW et al.; Lycopersicon species often contain the toxic glycoalkaloid alpha-tomatine, which is proposed to protect these plants from general microbial infection . however, fungal pathogens of tomato often are tolerant to alpha-tomatine and detoxification of alpha-tomatine may be how these pathogens avoid this potential barrier . As an initial step to evaluate this possibility, we have purfied to homogeneity a beta-1,2-D glucosidase from the tomato pathogen Septoria lycopersici that hydrolyzes the beta-1,2-D glucosyl bond on the tetrasaccharide moiety of alpha-tomatine to produce beta2-tomatine . The enzyme is a 110-kDa protein with a pI of 4.5 and a Km for alpha-tomatine of 62 microM . Little or no activity was detected on a variety of other glycosides . The gene encoding this protein was isolated and contains an open reading frame of 803 amino acids that shares sequence homology with several other beta-D-glucosidases . When S . lycopersici was incubated with alpha-tomatine, beta2-tomatinase mRNA accumulated, suggesting that the enzyme is substrate inducible . Aspergillus nidulans expressed inverted question markbeta2-tomatinase inverted question mark activity when transformed with this gene but transformants were still sensitive to alpha-tomatine. Scand J Clin Lab Invest, 1995 Nov, 55(7), 597 - 607 The human cystatin C gene promoter: functional analysis and identification of heterogeneous mRNA; Olafsson I; Human cystatin C is a low molecular weight protein involved in the control of human cysteine proteinase activity as well as microbial cysteine proteinase activity threatening the integrity of tissues . The gene for cystatin C is located on the short arm of chromosome 20, spans 6.5 kb and has three exons . To understand the mechanisms for the expression of cystatin C at the transcriptional level we mapped the 5' boundary of mRNA transcripts and studied the 5'-region of the cystatin C gene in a transient expression system with chimeric constructs utilizing various fragments of 1.1 kb of the 5'-flanking region coupled to the gene for human growth hormone . Mapping of the 5'-end of human cystatin C mRNA from placenta and seminal vesicles (low to medium versus high cystatin C expression, respectively) identified three major transcription initiation sites (positions -75, -78 and -80, A of initiation ATG as +1) and three minor sites (positions -98, -101 and -103) . The relative amounts of different mRNA species were approximately the same in these two tissues . Functional analysis of the 5'-region in cultured HeLa cells revealed one region (positions -279 to -156) with a strong positive effect on transcription and comprising three identical tandemly arranged GC-rich sequences. Adv Dent Res, 1995 Nov, 9(3), 239 - 43 Salivary factors in caries models; Hay DI; Consideration of salivary factors in caries models rarely extends beyond viewing saliva as a sink or diluent for plaque metabolic products, or as a source of buffering, for neutralizing plaque acids . In reality, saliva has a complex chemistry and a wide range of biochemical activities that may significantly affect plaque chemistry and microbiology . Thus, saliva is a major source of microbial nutrients, without which bacterial acid production is diminished . Buffering by salivary bicarbonate, and base production from urea and basic amino acids and peptides, significantly affect Stephan curves . Saliva is supersaturated with respect to basic calcium phosphate salts and contains novel inhibitors of calcium phosphate precipitation, while specific salivary proteins bind calcium . It seems important to consider if this system is reflected in plaque . Saliva, with contributions from serum and bacterial constituents, provides most of the precursors for the acquired enamel pellicle, which acts to slow demineralization during caries attack . Pellicle constituents appear to influence initial bacterial colonization of tooth surfaces and, therefore, may affect the microbial composition of plaque, but their detailed effects on plaque are poorly understood . Microbial adaptations to the anti-bacterial systems also seem important but are poorly investigated . Thus, saliva possesses an array of activities that have substantial actual or potential impact on plaque and, therefore, merit consideration for inclusion in systems intended to model dental caries. Poult Sci, 1995 Nov, 74(11), 1843 - 54 Response of turkey poults to tiered levels of Natuphos phytase added to soybean meal-based semi-purified diets containing three levels of nonphytate phosphorus; Ravindran V et al.; A 3-wk feeding trial using 920 day-old turkey poults was conducted to evaluate the addition of seven levels of phytase (Natuphos; 0, 200, 400, 600, 800, 1,000, and 1,200 U/kg of diet) to diets containing three levels of nonphytate P (nP) (.27, .36, and .45%) . A positive control diet contained .60% nP . Semi-purified basal diets contained soybean meal as the only protein source . The increase in BW gain from added phytase was greatest for the lowest nP diet (nP by phytase interaction, P < .001) . At .27% nP, gains improved (P < .001) to 800 U of phytase/kg of diet and then reached a plateau . At .36 and .45% nP, increases in gains were observed only for 200 U of phytase/kg of diet . The highest phytase addition to.36 and .45% nP diets produced gains equal to those of the positive control diet . Feed intake increases paralleled those of BW gains . Gain:feed was lowest for the .27% nP diets without phytase, but improved (P < .001) to 800 U of phytase/kg of diet and then reached a plateau . The high incidence of leg disorders and high mortality (40%) observed for the poults fed the .27% nP diet without added phytase declined with the addition of 200 to 400 U of phytase/kg of diet . Ash percentage of toes and tibias increased as the levels of nP (P < .001) and phytase (P < .01) increased; the magnitude of the response to phytase decreased as nP in the diet increased, resulting in an nP by phytase interaction (P < .001) . Tibial shear force and stress responded in a similar manner to increasing levels of nP and added phytase . Results show that 652 U of microbial phytase is equivalent to 1 g of P from defluorinated phosphate in turkey starter diets using soybean meal as the only source of phytate P . The response per 100 U of phytase decreased as the total amount of phytase added was increased. Poult Sci, 1995 Nov, 74(11), 1831 - 42 Improving phosphorus availability in soybean meal for broilers by supplemental phytase; Denbow DM et al.; A 21-d experiment was conducted with day-old male broilers (n=840) to evaluate the effectiveness of supplemental phytase for improving the availability of phytate P in soybean meal when varying levels of P were fed . The semi-purified basal diet (.18% phytate P) contained soybean meal as the only protein source . Seven levels of phytase (0, 200, 400, 600, 800, 1,000, and 1,200 U/kg diet) were added to diets formulated to contain .20, .27, or .34% nonphytate P (nP; or .38, .45, and .52% total P, respectively) . The desired levels of nP in the three basal P diets were achieved by adding varying amounts of defluorinated phosphate . A 2:1 Ca:total P ratio was maintained in all diets . Body weight gains and feed intake were improved (P < .001) by phytase at all nP levels, but the magnitude of response was greatest at low nP levels, resulting in an nP by phytase interaction (P < .01) . Gain:feed was unaffected by phytase addition . A high mortality (35 to 45%) was observed for the .20 and .27% nP diets without added phytase, but this declined to normal levels with the addition of 200 to 400 U phytase/kg diet . Ash percentage of toes and tibia and shear force and stress of tibia increased with added phytase . These responses clearly show that the phytate-bound P in soybean meal was made more available to broilers by microbial phytase, and the total response was related to the phytase and nP/total P levels . Based on the high R2 values for the second order translog equations, BW gain, feed intake, and toe ash percentage were the most sensitive indicators to assess P availability, followed by tibia force and ash percentage . Derived nonlinear and linear equations for BW gain and toe ash percentage at the two lower nP levels were used to calculate P equivalency values of phytase for inorganic P . Using the average function of P released ( gamma ) by microbial phytase ( chi ) derived with nP levels of .20 and .27% for BW gain and toe ash percentage, gamma = 1.120 - 1.102e-.0027chi, 1 g of P could be released with 821 U of phytase . The amount of P released increased with increasing levels of phytase, but the amount of P released per 100 U of phytase decreased . Released P ranged from 31 to 58% of phytate P for 250 to 1,000 U of phytase/kg of diet. Vet Immunol Immunopathol, 1995 Nov, 49(1-2), 1 - 13 Validation of immunoassays for bovine haptoglobin; Young CR et al.; Haptoglobin (Hp) is an acute phase protein . The plasma concentration of Hp increases rapidly following tissue damage associated with infection and inflammation . Thus Hp levels could be used as a screening test for organic disease, an objective index of disease activity and response to therapy, or as a sign of microbial infection . Recently, an enzyme-linked immunosorbent assay for bovine Hp was described . We have now developed three different immunoassay formats for bovine Hp and report on their validation and relative value to the diagnosis of bovine disease . Hp levels measured using these three immunoassays were compared and contrasted with results obtained for Hp estimation as measured by the increase in the protection of peroxidase activity against acid inactivation following binding with bovine haemoglobin . The quantitative Hp immunoassays evaluated in the present study are simple, rapid, inexpensive, reproducible, and well suited for both field and laboratory use. J Pharm Sci, 1995 Nov, 84(11), 1291 - 4 Effects of polyacrylic polymers on the lumenal proteolysis of peptide drugs in the colon; Bai JP et al.; The in-vitro effectiveness of polyacrylic acid polymers in inhibiting degradation of insulin, calcitonin, and insulin-like growth factor-I by colonic lumenal contents was determined . Further, the effect of Carbopol 974P, a polyacrylic acid polymer, on colonic absorption of insulin in rats was studied . The results revealed that Carbopol 934P, 971P, and 974P all strongly inhibited microbial proteolytic activities against insulin, calcitonin, and insulin-like growth factor-I . Inhibition by Carbopol polymers was complete or almost complete when the concentration of each polymer in saline or in 50 or 100 mM Tris buffer was 0.4%, where the pH of the medium was lower than 5 . The extensive inhibition by these polyacrylic acid polymers seems to correlate with their ability to acidify the incubation medium . Further, in-situ absorption studies showed that Carbopol 974P increased the pharmacological availability of colonic insulin . In summary, Carbopol polymers are useful in minimizing colonic proteolysis of peptide drugs. J Anim Sci, 1995 Nov, 73(11), 3476 - 84 Effect of barley variety and dietary barley content on digestive function in beef steers fed grass hay-based diets; Feng P et al.; Five ruminally and duodenally cannulated steers were used in a 5 x 5 Latin square design with a 2 x 2 + 1 arrangement of treatments to study the effects of barley variety and dietary barley content on digestive function in steers fed grass hay-based diets . Barley varities evaluated were Russell and Steptoe, which had bulk densities of 67.7 and 64.5 kg/hL, respectively . Supplemental treatments were as follows: corn, low Russell (Ru-lo), low Steptoe (St-lo), high Russell (Ru-hi), and high Steptoe (St-hi) . Corn, Ru-lo, and St-lo were provided at 30% (DM basis) of grass hay-based diets, whereas Ru-hi and St-hi were provided at 35.5% of diet DM (equal starch content as the corn treatment) . No treatment differences (P > .10) were observed for DMI, ruminal particulate passage rate, and NDF total tract digestibility . Ruminal DM and starch digestibility were greater (P < .01) for barley-containing diets than for the corn diet . Similarly, total tract DM (P < .10) and starch (P < .01) digestibility was greater for barley than for corn diets . Microbial protein and non-NH3 N flow to the small intestine were greater (P < .01) for the barley diets than for the corn diet . Starch intake (P < .01) and DM digestibility (P < .10) were greater for high- than for low-barley diets; however, differences due to barley variety were not observed (P > .10) . In situ disappearance of grass hay NDF at 8 and 96 h of incubation was greater (P < .05) for barley than for corn diets . Rate of in situ disappearance of grain DM was greater (P < .01) for barley than for corn and for Russell than for Steptoe barley . Responses suggest that ruminal and total tract digestibility and protein flow to the small intestine can be increased with barley compared with corn as an energy supplement to grass hay-based diets. J Anim Sci, 1995 Nov, 73(11), 3427 - 36 Supplemental protein and energy for beef cows consuming endophyte-infected tall fescue; Forcherio JC et al.; Effects of energy and protein supplementation of endophyte (Acremonium coenophialum)-infected (E+) and noninfected (E-) tall fescue (Festuca arundinacea Schreb.) on forage intake, digestibility, N flow to the small intestine, and cow-calf productivity was evaluated in two experiments . In Exp . 1, 10 ruminally and duodenally cannulated steers were fed either E- or E+ hay with four supplements or E- or E+ hay unsupplemented . Four supplements formulated with either cracked corn or soybean hulls with 100 or 200 g/d of ruminally undegraded intake protein (UIP) were compared . Levels of UIP were varied by adding soybean meal or blood meal . Hay OM intake was not affected (P > .20) by source of energy of level of UIP; however, intake of E- was greater (P < .05) than that of E+ . True ruminal OM digestion tended to be greater (P < .12) for steers fed 200 g/d of UIP than for steers fed 100 g/d . Steers receiving 200 g/d of UIP had increased (P < .10) total N flow to the duodenum compared with steers receiving 100 g/d but similar (P > .20) microbial efficiencies . In Exp . 2, 30 cows (average initial BW 459 +/- 26 kg) and their calves (average initial BW 74 +/- 5 kg and 74 +/- 5 d of age) grazed an 8.1-ha E+ pasture from late May to late July . Cows were individually fed supplements used in Exp . 1 each day . Cows that received cracked corn lost .10 kg/d when fed 100 g/d of UIP but gained .33 kg/d when fed 200 g/d . Cows fed soybean hulls and 100 g/d of UIP gained .07 kg/d, whereas cows provided 200 g/d lost .10 kg/d . Calves nursing cows supplemented with 100 g/d of UIP gained more (P < .08) BW because of increased (P < .07) milk consumption and slightly greater (P < .19) forage intake than calves nursing cows supplemented with 200 g/d of UIP. Hokkaido Igaku Zasshi, 1995 Nov, 70(6), 839 - 46 Vaginal and cervical pH in bacterial vaginosis and cervicitis during pregnancy; Sagawa T et al.; The purpose of our study is to see whether vaginal and cervical pH are helpful to screen for bacterial vaginosis and cervicitis during pregnancy . One hundred and seven pregnant women underwent prospectively both vaginal pH and cervical pH measurement and screening for microbial infections of the lower genital tracts at 10, 20 and 30 gestational week between February 1993 and August 1993 . The value of vaginal pH significantly elevated in bacterial vaginosis (P < 0.05) . Vaginal pH values also significantly elevated in patients who had had sexual intercourse 2 days before the dates of examinations (P < 0.01) . Cervical pH value exhibited no significant change in bacterial vaginosis and cervicitis . Vaginal pH is a valid indicator for screening of bacterial vaginosis during pregnancy . However, vaginal pH might be influenced by the presence of semen . Cervical pH is not a useful parameter for screening of cervicitis during pregnancy. Ann Plast Surg, 1995 Nov, 35(5), 455 - 7 Clinical significance of intracapsular fluid in patients' breast implants; Ahn CY et al.; Clinical reports on the incidence and clinical significance of intracapsular fluid are lacking in the literature . It remains unknown whether the presence of intracapsular fluid has any relation to implant infection or colonization . The purpose of this study was to determine the frequency and type of intracapsular fluid, specifically, whether intracapsular fluid causes implant infection, implant rupture, or bacterial colonization . A total of 139 implants from 72 symptomatic patients were entered into the prospective clinical study . Our study demonstrated the presence of intracapsular fluid in 21 of 139 (15%) implants . Positive microbial cultures were identified in 39% of the implants in the positive intracapsular fluid group, compared to 43% in the negative fluid group . There was no statistically significant difference between these groups . Also, no adverse clinical relationship was demonstrated between local symptoms and presence of intracapsular fluid . There was, however, a positive trend toward the presence of fluid when implant shell types were nonsmooth (polyurethane and textured silicone implants) . Further studies are indicated to elucidate the fluid production mechanism and possible secretory activity of prosthetic capsules interfacing the textured breast implant surface. J Clin Microbiol, 1995 Nov, 33(11), 2927 - 30 Enzyme-linked immunosorbent assay-based inhibition test for neutralizing antibodies to polioviruses as an alternative to the neutralization test in tissue culture; Edevag G et al.; A poliovirus-binding inhibition test (PoBI test) was established for the quantitative determination of antibodies to polioviruses and was evaluated in comparison with the conventional neutralization test (NT) . The first step of the PoBI test is an incubation of serial dilutions of test samples with inactivated poliovirus followed by the detection of free viral epitopes by a double antibody sandwich enzyme-linked immunosorbent assay with type-specific capture polyclonal antisera and type-specific neutralizing monoclonal indicator antibodies . A comparison of the PoBI test with the conventional NT for antibodies to all three types in 100 human serum samples showed excellent correlations (r > 0.95) over a wide range of antibody concentrations . The PoBI test, not necessitating live virus and tissue culture facilities, could be a simple alternative to the NT, and the principle of the assay is potentially applicable to other microbial systems. J Clin Immunol, 1995 Nov, 15(6), 284 - 92 CpG DNA: a pathogenic factor in systemic lupus erythematosus? Krieg AM. Systemic lupus erythematosus (SLE) is a multifactorial disease of unknown etiology . Characteristic features of SLE include (1) polyclonal B cell activation, (2) overexpression of the immune stimulatory cytokine interleukin-6 (IL-6), (3) defective tolerance to self antigens, and (4) production of anti-DNA antibodies (Ab) . Bacterial infection has been suspected as a triggering factor for lupus . Bacterial DNA differs from vertebrate DNA in the frequency and methylation of CpG dinucleotides . These CpG motifs in bacterial DNA induce a variety of immune effects, including (1) polyclonal activation of murine and human B cells, (2) IL-6 secretion, and (3) resistance to apoptosis, thereby potentially allowing the survival of autoreactive cells . These results suggest that microbial DNA could therefore be a pathogenic factor in SLE . SLE patients have elevated levels of circulating plasma DNA which is reportedly enriched in hypomethylated CpGs . Genomic DNA is also hypomethylated in SLE . The purpose of this review is to summarize the immune effects of CpG motifs and to present the evidence for their possible involvement in the pathogenesis of SLE. Int J Food Sci Nutr, 1995 Nov, 46(4), 319 - 25 Irradiated versus fumigated spices in sausage; Bolander CR et al.; The efficacy of gamma radiation and ethylene oxide fumigation as decontaminating treatments for spices used in sausages was compared . Microbial loads of sausages containing irradiated, fumigated or non-treated spices were examined over three intervals of time . No significant differences between microbial loads of Mexican sausages containing irradiated spices (10 kGy) verses ethylene oxide-treated spices were found . Mexican sausages containing treated spices had significantly lower yeast, mould and spore counts than control sausages . After 8 weeks of storage, only the microbial loads of control sausages suggested spoilage . No discernable differences in treatment efficacy were noted with respect to yeast, mould and spore counts in Italian sausages. Yakugaku Zasshi, 1995 Nov, 115(11), 892 - 908 {Biosynthesis and microbial transformation of griseofulvin and carcinogenesis and prevention of cancer by estrogens}; Sato Y; In the biosynthetic study of griseofulvin by Penicillium urticae and microbial transformation of (-)- and (+)-dehydrogriseofulvin and their derivatives by Streptomyces cinereocrocatus excellent informations were obtained from 2H-NMR spectroscopy . In the reduction of (-)-dehydrogriseofulvin into (+)-griseofulvin by a partially purified enzyme system of S . cinereocrocatus, the origin of the 6' alpha-hydrogen of (+)-griseofulvin was a hydride ion donated by pro-4R-hydrogen of NADPH . In connection with the study of carcinogenesis, diethylstilbestrol (DES) was proved to disrupt microtubules in vitro . The other synthetic estrogens, E,E-dienestrol, meso-hexestrol, and dl-hexestrol were inhibitors of microtubule assembly in vitro, and induced twisted ribbon structures or ribbon-sheet-microtubules from microtubule proteins . Next, the effects of DES and its methyl ethers on the chromosome of and the cellular microtubule architecture, revealed by fluorescent anti-tubulin antibody, of Chinese hamster V79 cells were examined, and further estradiol-17 beta was proved to exhibit higher microtubule-disruptive activity than DES in V79 cells . Furthermore, cytoplasmic microtubules in the human breast cancer cell lines MCF-7 and MDA-MB-231, estrogen receptor-positive and -negative cell lines, respectively, were disrupted equally by estradiol-17 beta . Then, natural estrogens and their derivatives comprising 30 compounds in total were tested in Chinese hamster V79 cells, proving that 2-methoxyestradiol showed the strongest activity (EC50: 2 microM) to disrupt microtubules . Further, in the assay of indenestrol A, a metabolite of DES, indenestrol B and their monomethyl ethers, the 4'-methyl ether of {(-)-3S}-indenestrol B exhibited both the strongest cytotoxicity in, and greatest disruption of the cellular microtubules of V79 cells, and no correlation with the affinity for estrogen receptors was shown. Laryngorhinootologie, 1995 Nov, 74(11), 684 - 90 {Establishment and characterization of 2 cell lines of squamous epithelial carcinoma of the mouth floor and tongue}; Gorogh T et al.; OBJECTIVE: The cultivation of squamous cell carcinoma cells of the upper aerodigestive tract is more difficult compared to other malignancies because of microbial contamination . The establishment of new squamous cell carcinoma cell lines is absolutely necessary to further characterize this tumor entity . METHODS: Two squamous cell carcinoma cell lines of the floor of the mouth and the tongue were established from tumor biopsies . They were examined by light and electron microscopy . Furthermore immunocytochemical and DNA cytofluorometric studies were performed . The growth features were tested in a tumor-stem cell assay and through transplantation in thymusless nude mice . RESULTS: The epithelial character of the cells was determined morphologically, immunocytochemically, and immunologically . Both cell lines shaved an aneuploid DNA Index of 1.92 for the tongue carcinoma cell line (HCTK-1) and 1.77 for the floor of the mouth cell line (HCFMK-1) . The plating efficiency was 0.008 and 0.01% . Cell differentiation in the original tumor and the nude mice tumor was identical . HCTK-1 is in the 25th passage and HCFMK-1 in the 42nd . CONCLUSIONS: The two new squamous cell carcinoma cell lines HCFMK-1 and HCTK-1 may serve as a basis for various in vitro investigations and may help to further characterize squamous cell carcinomas of the upper aerodigestive tract. Microbiology, 1995 Nov, 141 ( Pt 11), 2897 - 904 Arylsulphatase from Alteromonas carrageenovora; Barbeyron T et al.; Arylsulphatase activity was identified in cultures of the marine bacterium Alteromonas carrageenovora, using methylumbelliferyl sulphate as substrate . In contrast with most other microbial arylsulphatases, arylsulphatase production in A . carrageenovora was not repressed by sulphate . The structural gene of arylsulphatase (atsA) was cloned and sequenced . An ORF of 984 bp was found, specifying a primary translation product of 328 amino acids with a molecular mass of 35797 Da . Arylsulphatase was partially purified from cell extracts of both A . carrageenovora and recombinant Escherichia coli . Both the recombinant and native enzymes exhibited a pI of 5.5, a Michaelis constant for methylumbelliferyl sulphate of 68 microM, and a molecular mass of approximately 35,000 Da in SDS-PAGE analysis . Secondary structure comparisons using hydrophobic cluster analysis suggest functional analogies between the arylsulphatase of A . carrageenovora, that of Mycobacterium leprae and a 33.5 kDa protein from Porphyromonas gingivalis . It is speculated that these proteins are all glycosulphohydrolases, involved with desulphatation of sulphated polysaccharides. Natl Med J India, 1995 Nov-Dec, 8(6), 261 - 2 Value of antibiotic therapy in tuberculin-positive children with parenchymal lung lesions; Somu N et al.; BACKGROUND . A tuberculin-positive child with radiological evidence of a parenchymal lung lesion is likely to be treated for tuberculosis by a physician . However, non-tuberculous microbial infections may also cause parenchymal lung lesions . We tried to distinguish tuberculous from non-tuberculous lung lesions by administering a course of antibiotics . METHODS . Three hundred and five tuberculin-positive children with parenchymal lung lesions due to pneumonia, bronchiectasis (cylindrical and reversible) and minor fissure opacification were studied at the Tuberculosis Clinic, Institute of Child Health, Madras . Those with more serious forms of tuberculosis like miliary, cavitary and segmental lesions and with grade III and IV undernutrition were excluded . Three weeks of oral antibiotic therapy, with erythromycin (30 mg/kg/day) and chloramphenicol (50 mg/kg/day) for the first two weeks followed by co-trimoxazole (trimethoprim 6 mg/kg/day and sulphamethoxazole 25 mg/kg/day) for the third week, was given . Chest X-rays were taken before and after antibiotic therapy . RESULTS . Sixty per cent of the children with pneumonia, 57% with bronchiectasis and 62% with minor fissure opacification showed complete radiological clearance . CONCLUSION . In tuberculin-positive children with parenchymal lung lesions radiological clearance was seen in 60% after three weeks of antibiotic therapy indicating that the parenchymal lung lesions were caused by non-tuberculous organisms . Hence a course of antibiotic therapy in these children may have diagnostic value as well as considerable financial, social and therapeutic implications. J Exp Med, 1995 Nov 1, 182(5), 1557 - 65 CD40 ligation induces Apo-1/Fas expression on human B lymphocytes and facilitates apoptosis through the Apo-1/Fas pathway; Schattner EJ et al.; The Apo-1/Fas antigen (CD95) mediates programmed cell death of lymphocytes when bound by Fas ligand or anti-Apo-1/Fas antibody . In contrast, the CD40 antigen provides a potent activation and survival signal to B lymphocytes when it is engaged by its T cell ligand (CD40L, gp39) or cross-linked by anti-CD40 antibody . In this study, we use human tonsillar B cells and the Ramos Burkitt's lymphoma B cell line, which serves as a model for human germinal center B lymphocytes, to study the effectors of Apo-1/Fas expression and apoptosis of human B cells . We found that Apo-1/Fas expression was upregulated on both malignant and normal human B lymphocytes after CD40 ligation induced by (a) cognate T helper-B cell interaction mediated by microbial superantigen (SAg); (b) contact-dependent interaction with CD40L+, but not CD40L- Jurkat mutant T cell clones; and (c) monoclonal anti-CD40, but not any of a panel of control antibodies . Enhanced B cell Fas/Apo-1 expression is functionally significant . Coculture of Ramos Burkitt's lymphoma line cells with irradiated SAg-reactive CD4+ T cells with SAg or CD40L+ Jurkat T cells results in B cell apoptosis, evidenced by reduced cell viability and DNA laddering . This process is augmented by the addition of anti-Apo-1/Fas monoclonal antibody, consistent with an acquired susceptibility to Apo-1/Fas-mediated apoptosis . These data support an immunoregulatory pathway in which seemingly contradictory signals involving the B cell proliferation/survival antigen CD40, as well as the Apo-1/Fas molecule, which mediates programmed cell death of lymphocytes, are linked in the process of human B cell activation. J Bone Joint Surg Br, 1995 Nov, 77(6), 967 - 70 A new in-vitro model to investigate antibiotic penetration of the intervertebral disc; Thomas Rde W et al.; We have studied the ability of a range of antibiotics to penetrate intervertebral disc tissue in vitro, using a mouse disc model . Equilibrium concentrations of antibiotics incorporated into the entire disc were determined by bioassay using a microbial growth-inhibition method . Uptake was significantly higher with positively-charged aminoglycosides compared with negatively-charged penicillins and cephalosporins . Uncharged ciprofloxacin showed an intermediate degree of uptake . Our results support the hypothesis that electrostatic interaction between charged antibiotics and negatively-charged glycosaminoglycans in the disc is an important factor in antibiotic penetration, and may explain their differential uptake. Fertil Steril, 1995 Nov, 64(5), 1012 - 22 Screening for subclinical inflammation in ejaculates; Eggert-Kruse W et al.; OBJECTIVE: To determine the clinical significance of albumin determination in ejaculates by means of an easy office test to screen semen samples for subclinical infection-inflammation . PATIENTS: One hundred fifty-nine randomly chosen males of couples with longstanding infertility (median duration of infertility 4 years (range 1 to 19 years) without clinical signs or symptoms of genital tract infection . SETTING: Outpatient Infertility Clinic of the University of Heidelberg, Germany . MAIN OUTCOME MEASURES: Screening of ejaculates for subclinical infection-inflammation by means of a ready-to-use kit for semiquantitative detection of albumin in addition to determination of leukocytes rates by means of monoclonal antibodies for differentiation of round cells and measurement of granulocyte elastase concentration in semen samples . Evaluation of sperm quality by means of standard sperm analysis including determination of local antisperm antibodies with the mixed antiglobulin reaction, evaluation of sperm functional capacity in vitro with the standardized sperm-cervical mucus (CM) penetration test, and semen cultures . All tests were performed from aliquots of the same ejaculates . RESULTS: Screening of semen samples for elevated albumin with the modified paper strips proved to be very easy, quick, and suitable for routine use . Positive results were not related markedly to medical history and outcome of clinical examination as well as to standard parameters of sperm analysis and were not influenced by local antisperm antibodies of the immunoglobulin (Ig)G and/or IgA class and microbial colonization . However, albumin-positive semen samples were significantly less frequent in case of very good outcome of the sperm-CM penetration test . A significant relationship was found with high rates of leukocytes of the round cells in semen samples (total range 0% to 96%) and the concentration of granulocyte elastase (total range 1 to 880 micrograms/L) . CONCLUSIONS: The results of this prospective study suggest that the determination of albumin in semen samples with ready-to-use test kits might be a valuable additional marker for subclinical infection-inflammation of the male genital tract and therefore suitable for screening during infertility investigation. Plast Reconstr Surg, 1995 Nov, 96(6), 1430 - 4 Paecilomyces variotii contamination in the lumen of a saline-filled breast implant; Young VL et al.; This report describes a case of gross contamination with the filamentous fungus P . variotii cultured from an intraluminal saline breast implant removed from a patient 14 months after implantation because of severe capsular contracture . We suspect the fungal contamination occurred when a container of saline was left open in the operating room prior to filling and placement of the implant . This case may be the first documented report of microbial growth and reproduction in the internal environment of a saline implant . We assume that organisms such as P . variotii can survive--and accumulate biomass--on the minute amounts of substrates that diffuse across an implant envelope. J Nutr, 1995 Nov, 125(11), 2751 - 62 Pyridoxine and pyridoxine-5'-beta-D-glucoside exert different effects on tissue B-6 vitamers but similar effects on beta-glucosidase activity in rats; Nakano H et al.; Pyridoxine glucoside is a partially available form of vitamin B-6 present in plant-derived foods . In this set of three studies, rats were fed diets containing different concentrations of either pyridoxine or pyridoxine glucoside in the presence or absence of a fixed concentration of pyridoxine for 2 wk . The distribution of B-6 vitamers and beta-glucosidase activity in tissues was examined to determine the metabolic effects of chronic consumption of pyridoxine glucoside . Rats fed pyridoxine glucoside either with or without pyridoxine exhibited a significant increase in the amount of hepatic pyridoxine 5'-phosphate (not detected in rats fed pyridoxine alone), whereas hepatic pyridoxal 5'-phosphate was decreased with increasing dietary pyridoxine glucoside . The activity of cytosolic beta-glucosidases in small intestine and kidney was affected by the dietary concentration of both pyridoxine and pyridoxine glucoside . Enzymatic activity capable of hydrolyzing pyridoxine glucoside was found in mucosal and intraluminal fractions of small intestine and in the kidney . Other tissues examined, including liver, spleen and stomach, did not hydrolyze pyridoxine glucoside in detectable quantities . These findings indicate that microbial and mucosal enzymes can participate in the intestinal hydrolysis of pyridoxine glucoside and that the kidney may contribute to postabsorptive hydrolysis . These findings further support the observations that dietary pyridoxine glucoside influences vitamin B-6 metabolism. Biochim Biophys Acta, 1995 Oct 26, 1259(1), 73 - 81 Preparative-scale kinetic resolutions catalysed by microbial lipases immobilised in AOT-stabilised microemulsion-based organogels: cryoenzymology as a tool for improving enantioselectivity; Rees GD et al.; Gelatin-containing microemulsion based organogels have been used as an immobilisation matrix for lipases from a number of different sources . Kinetic resolutions of octan-3-ol, 1-octen-3-ol and 1-octyn-3-ol by esterification with decanoic acid have been performed using Chromobacterium viscosum (CV) lipase . CV lipase is highly enantioselective in favour of the (R)-(-) isomer of octan-3-ol, but the enantioselectivity is both reversed and decreased by the introduction of unsaturation at the 1-position . Marked improvements in enantioselectivity were achieved by carrying out the reaction at -15 degrees C, the enantiomeric excess of the ester product increasing from 47% (E = 3) to 73% (E = 8) in the case of 1-octen-3-ol, and from 17% (E = 1.4) to 38% (E = 2.5) in the case of 1-octyn-3-ol . The enantiomeric excess was approximately 85% (E approximately 15) for octan-3-ol, and there was no marked improvement in enantioselectivity even at -15 degrees C . Apparent activation energies for the esterification using decanoic acid of octan-3-ol, 1-octen-3-ol and 1-octyn-3-ol by CV lipase were 32 kJ mol-1, 31 kJ mol-1 and 41 kJ mol-1, respectively . This compares to an activation energy of 21 kJ mol-1 for the esterification of octan-1-ol with decanoic acid using CV lipase under the same conditions . Lipases from Pseudomonas (Fluka), Pseudomonas (Genzyme) and lipoprotein lipase ex Microbial (Genzyme) also selectively esterified the (R)-(-) isomer of racemic octan-3-ol, the two Pseudomonas preparations yielding product with an enantiomeric excess of 90% . Candida cylindracea lipase did not exhibit activity in gelatin-containing MBGs . Large-scale syntheses were performed in a 1 dm3 batch reactor in which 200 cm3 of pelleted MBG (containing 350 mg of CV lipase) was used repeatedly for the kinetic resolution of octan-3-ol. Biochim Biophys Acta, 1995 Oct 25, 1252(2), 305 - 11 Molecular characterization of a Galactomyces geotrichum lipase, another member of the cholinesterase/lipase family; Phillips A et al.; Geotrichum candidum secretes several lipase isoenzymes, differing in their selectively towards esters of long chain fatty acids with a cis-9 double bond . One group shows an absolute selectively towards these fatty acid esters, the other group has a more relaxed specificity and will also hydrolyze other long chain fatty acid esters . Galactomyces geotrichum secrets a lipase that has the same specificity as the latter group . The corresponding lipase gene was cloned from Galactomyces geotrichum . From an alignment of our enzymes' primary structure with those of different strains of Geotrichum candidum, remarkable conservation is evident and it is not yet possible to identify residues/structures responsible for differences in fatty acid specificity . Comparison of the GCL/GGL family with a variety of lipases from other sources, indicated that they are more related to mammalian than microbial lipases. J Biotechnol, 1995 Oct 16, 42(3), 271 - 80 Solution and properties of age population balance models which assume discrete division ages; Hjortso M; A solution procedure for models of the transient age distribution which employ the assumption of discrete, constant division ages is developed . The solutions for the final state of the age distribution, the solutions obtained in the limit as time goes to infinity, are found to have properties which do not make biological sense . In particular, the solutions will only approach the steady-state solution for very special initial conditions . For most initial conditions, the solution for the final state will instead exhibit an oscillatory behavior . In addition, the oscillatory solutions are unstable with respect to changes in values of the model parameters and solutions with very different periods of oscillation are found arbitrarily close to each other in the parameter space . Models which assume discrete division ages must therefore be used with caution and may be unsuitable as models of autonomous microbial oscillations. Ophthalmology, 1995 Oct, 102(10), 1476 - 85 Cytologic evidence of corneal diseases with limbal stem cell deficiency; Puangsricharern V et al.; PURPOSE: To determine which human corneal diseases show similar abnormal corneal surfaces, characterized by conjunctival epithelial ingrowth (conjunctivalization), vascularization, and chronic keratitis (i.e., a constellation of signs termed limbal stem cell dysfunction {deficiency}, which have been noted in experimental rabbit models) . METHODS: A total of 134 impression cytology specimens of the perilimbal region collected from 1984 to 1994 were reviewed . Limbal deficiency was diagnosed if conjunctival goblet cells were found on the corneal surface . RESULTS: Ninety-four patients were found to have limbal deficiency . Category 1 comprised 53 patients with a clear history showing limbal stem cell destruction by chemical/thermal burns, Stevens-Johnson syndrome, multiple surgeries and cryotherapies, contact lens wear, and severe microbial keratitis . Patients in category 2 (n = 41), did not have such a history, but gradual loss of stem cell functions over time was disclosed and included diverse causes such as aniridia, multiple endocrine deficiencies, neurotrophic keratopathy, peripheral inflammatory keratopathy or limbitis, and idiopathy . The 40 remaining patients with suspicious findings did not have limbal deficiency . CONCLUSIONS: Impression cytology can be used to diagnose and monitor corneal diseases with limbal deficiency, which manifest distinct clinical problems and are generally poor candidates for penetrating keratoplasty . The identification of category 1 diseases allows one to consider limbal (stem cell) transplantation for surface reconstruction . The presence of category 2 diseases indicates that limbal stem cell functions can be modulated by developmental, hormonal, neuronal, vascular, and inflammatory factors in the limbal stroma. Int J Immunopharmacol, 1995 Oct, 17(10), 793 - 803 The specific type IV phosphodiesterase inhibitor rolipram combined with adenosine reduces tumor necrosis factor-alpha-primed neutrophil oxidative activity; Sullivan GW et al.; Monocytes and macrophages produce tumor necrosis factor-alpha (TNF alpha) in response to microbial products including endotoxin . TNF alpha is a potent primer of neutrophil (PMN) oxidative activity . Certain xanthine phosphodiesterase (PDE) inhibitors such as pentoxifylline have been shown to inhibit stimulated oxidative activity in PMN . In the present study, the non-xanthine PDE type IV inhibitor rolipram (4-{3'-cyclopentyloxy-4'-methoxyphenyl}-2-pyrrolidone) alone and in combination with adenosine is examined as a potential modulator of TNF alpha-primed PMN oxidative activity . Attainable in vivo concentrations of rolipram and physiological concentrations of adenosine alone and together synergistically decreased rhTNF alpha-primed suspended PMN oxidative activity stimulated by the chemoattractant f-met-leu-phe . The rolipram effect was reversible by washing, and rolipram had a comparable effect if added before or after priming, indicating that its effect was on the primed response rather than on priming per se . In addition, rolipram especially when combined with adenosine, decreased rhTNF alpha-stimulated PMN adherence to a fibrinogen-coated surface, and the oxidative burst of rhTNF alpha-stimulated adherent PMN . The specific adenosine A2a receptor agonists CGS 21680 and WRC-0474 had comparable activity to adenosine in these experiments . Adenosine (or CGS 21680) combined with rolipram synergistically increased f-met-leu-phe-stimulated PMN cAMP content . The effects of both adenosine and rolipram with adenosine could be only partly counteracted by treatment of the PMN with the protein kinase A inhibitor KT 5720, indicating that protein phosphorylation is only partially involved . Rolipram activity was about 1000 x (by molar concentration) greater than pentoxifylline in comparable assays . Thus, rolipram, especially when combined with adenosine, has potent modulating effects on PMN activation and may be useful in decreasing inflammatory tissue damage in patients with sepsis. New Microbiol, 1995 Oct, 18(4), 409 - 22 Structural differences of the microbial community in two sites of the Terra Nova Bay (Ross Sea, Antarctica): a statistical analysis; Bruni V et al.; The microbial community structure was investigated in a one month survey at two sampling stations located in the Terra Nova Bay (Antarctica) . The microbial assemblage was studied by direct counts with epifluorescent microscopy (picoplankton and picophytoplankton), fractionated adenosine-triphosphate, lipopolysaccarides and bacterial growth on cultural medium . The results underwent the Spearman rank correlation and the principal component analysis together with other environmental parameters (temperature, salinity, nutrient salts and chlorophyll a) with the aim of revealing differences in the microbial community structure between the in-shore and off-shore sites. Pharm Res, 1995 Oct, 12(10), 1493 - 8 Microbial and mammalian metabolism studies on the semisynthetic antimalarial, deoxoartemisinin; Khalifa SI et al.; PURPOSE . Deoxoartemisinin is a semisynthetic antimalarial with potential for treatment of multiple drug resistant malaria . Metabolism studies were conducted to aid in future drug development . METHODS . Microbial model systems were employed which have been shown to be good predictors of mammalian drug metabolites . Metabolism studies using rats were also performed . RESULTS . Three microbial metabolites of deoxoartemisinin were identified (2, 3, and 4) . Metabolite 3 was also found in rat plasma . HPLC/MS analyses were performed on the rat plasma using 2, 3, and 4 as standards . All metabolites were thoroughly characterized by 1H and 13C-NMR . An additional rat plasma metabolite was revealed and it was shown not to be 9 alpha-hydroxyartemisinin . CONCLUSIONS . Deoxoartemisinin was metabolized to three microbial metabolites . Metabolism by rats showed the presence of two metabolites in the plasma, one of which was the same as the microbial metabolite. Int J Food Microbiol, 1995 Oct, 27(2-3), 139 - 46 Accuracy of microbial growth predictions with square root and polynomial models; Delignette-Muller ML et al.; The results of growth predictions using square root and polynomial models published in 14 papers were studied . Errors on quantities of practical interest such as lag time, generation time or the time required to reach a given increase in number of cells, are analyzed . The distribution of these errors was examined with the perspective of the practical use of predictive models in food industry . Highly unsafe predictions and significant average errors were observed in some cases . A good knowledge of predictive models accuracy seems essential for their efficient and safe use, for example to predict the shelf life of a product . Yet, authors generally gave no pragmatic information on such things as the average relative error or the range of errors on predicted variables . Problems of robustness of models when tested in different conditions were noticed, which corroborates the necessity of a systematic validation of models on new data. Int J Food Microbiol, 1995 Oct, 27(2-3), 107 - 15 The application of the WLF equation to predict lag time as a function of temperature for three psychrotrophic bacteria; Schaffner DW; Many mathematical models for microbial growth rates or lag times have been proposed . Most of these models either predict well, but are empirical and offer no insight into mechanism or they reflect mechanism but are too complex or do not fit the data adequately . The Williams-Landel-Ferry (WLF) equation (used by polymer chemists) is empirical, but shows some relation to first principles, and hence may offer some insight into mechanism . This model also has only three parameters and therefore a fairly simple form . The WLF equation was fit to lag times derived from three datasets previously developed in our laboratory . The fits obtained with the WLF equation were as good as the best fits obtained with other models (e.g Arrhenius, Davey, response surface and square root) . The WLF model was able to account for 98 to 99% of the variance in the three datasets, indicating a very good fit overall . The parameter estimates of the WLF model were not as highly correlated as those of some of the other models . Many of the models, including the WLF equation did not predict well at very long lag times . Weighted least squares non-linear regression improved the fit for these long lag times. Curr Opin Struct Biol, 1995 Oct, 5(5), 622 - 35 Microbial recognition of target-cell glycoconjugates; Karlsson KA; Attachment of microbial protein to host cell-surface carbohydrate is considered essential for successful infection . Information of conceptual interest, including crystal structures of protein-saccharide complexes and convincing models of target mem