J Bacteriol, 1992 Aug, 174(15), 5021 - 6 Light and oxygen regulation of the synthesis of bacteriochlorophylls a and c in Chloroflexus aurantiacus; Oelze J; Control of the synthesis of bacteriochlorophylls (Bchls) a and c by light and oxygen was studied in Chloroflexus aurantiacus grown in batch or chemostat culture with serine as the growth-limiting substrate . For comparison, inhibition by gabaculine of the formation of selected tetrapyrroles was studied . The inhibitory effect of gabaculine decreased in the following order of tetrapyrrole formation: coproporphyrin greater than Bchl c greater than Bchl a . Not only did addition of 5-aminolevulinate (ALA) reverse the inhibition by gabaculine, it also caused an increase in Bchl c content when the cultures grew at high concentrations of ALA . Inhibition of Bchl a, Bchl c, and coproporphyrin formation by oxygen was similar to inhibition by gabaculine . Addition of ALA to aerated cultures led to significant accumulation of coproporphyrin . These results suggest that oxygen inhibits tetrapyrrole formation at a site before ALA formation . Control by light was studied with chemostat cultures transferred from 5 klx to 25 klx . This resulted in only a transient increase of the protein level of the culture, while specific contents of Bchls c and a and the ratio Bchl c/Bchl a decreased to lower steady states . However, the specific content of coproporphyrin increased . Addition of ALA to chemostat cultures adapted to 50 klx increased specific coproporphyrin and Bchl c contents by factors of about 20 and 4, respectively, while the specific Bchl a content was only slightly increased and protein levels were unaffected . Increasing the serine concentration caused an initial increase in the specific Bchl c content, which returned to the original value as soon as the protein content had attained its maximal level . These results suggest that light does not control ALA formation as strictly as oxygen and that competition of biomass formation and tetrapyrrole synthesis for common precursors may be influenced by light. J Gen Microbiol, 1992 Aug, 138 ( Pt 8), 1671 - 7 Demonstration of the intracellular production of tissue-destructive protease by Legionella pneumophila multiplying within guinea-pig and human alveolar macrophages; Rechnitzer C et al.; The major extracellular enzyme of Legionella pneumophila, a metalloprotease, has been proposed as a pathogenic factor in Legionnaires' disease due to its cytotoxic, tissue-destructive, and phagocyte-inhibitory properties . The relevance of these activities depends on the production of the protease during infection, i.e . by L . pneumophila multiplying intracellularly . In this study, L . pneumophila was demonstrated to produce protease in guinea-pig and human alveolar macrophages infected in vitro . After 24 h infection, approximately 0.1 to 0.2 micrograms of protease per 10(6) bacteria was measured by ELISA in culture supernatants and lysates of the infected cells, whereas no protease could be detected immediately after infection . Immunogold labelling using anti-protease antibody showed the enzyme to be located within phagosomes and distributed throughout the macrophages . Recent observations have shown that this protease could modify host defence mechanisms through inhibition of bacterial killing by neutrophils and monocytes . The intracellular production of the enzyme in infected macrophages demonstrated here further supports a role for the protease in the pathogenesis of Legionnaires' disease. J Gen Microbiol, 1992 Aug, 138 ( Pt 8), 1613 - 21 An iron stress operon involved in photosynthetic electron transport in the marine cyanobacterium Synechococcus sp . PCC 7002; Leonhardt K et al.; The iron-stress-induced genes isiA and isiB have been cloned and sequenced from the marine unicellular cyanobacterium Synechococcus sp . PCC 7002 . These genes code for a photosystem II chlorophyll-binding protein and flavodoxin respectively . The genes form a dicistronic operon that is transcriptionally activated under iron-stress conditions to produce an abundant monocistronic message containing isiA and a much less abundant dicistronic message that also contains isiB . The arrangement of these genes, their transcriptional control and the relative abundance of the monocistronic and dicistronic messages produced under iron stress parallels the pattern shown by the freshwater cyanobacterium Synechococcus sp . PCC 7942 . The genes for the corresponding proteins found under iron-replete conditions, CP-43 and ferredoxin, have also been cloned and sequenced . Northern blot analysis indicates that both of these genes are constitutively expressed under both iron-stress and iron-replete conditions. Am J Infect Control, 1992 Aug, 20(4), 177 - 84 Aerosol penetration through surgical masks; Chen CC et al.; BACKGROUND: Surgical masks are used in hospitals to reduce postoperative infection in patients . The presence of aerosols containing pathogens makes it desirable to protect the medical staff as well . METHODS: The collection efficiencies of surgical masks measured with two aerosol-size spectrometers . The flow rates through the masks were varied from 5 to 100 L/min to study the flow dependency . For comparison, several industrial-type respirators were also tested . RESULTS: A surgical mask consisting of filter material performed better than did a surgical mask consisting only of a shell with a coarse pore structure . The latter passed 80% of submicrometer-sized aerosols with little flow dependency, whereas the penetration of submicrometer-sized aerosols through the mask made of filter material ranged from 25% at a flow rate of 5 L/min to 70% at 100 L/min . CONCLUSIONS: The mask that has the highest collection efficiency is not necessarily the best mask from the perspective of the filter-quality factor, which considers not only the capture efficiency but also the air resistance . Although surgical mask media may be adequate to remove bacteria exhaled or expelled by health care workers, they may not be sufficient to remove the submicrometer-size aerosols containing pathogens to which these health care workers are potentially exposed. Poult Sci, 1992 Aug, 71(8), 1391 - 5 A rapid method for the determination of temperature abuse of fresh broiler chicken; Russell SM et al.; Ready-to-cook broiler chicken carcasses were obtained immediately postchill from a commercial processing plant . A control group was held at 4 C for 10 days . Test groups were held at 4 C for 2 days, temperature abused at 10, 15, or 20 C for 0, 1, 2, 4, 6, 8, 12, or 24 h, and then held at 4 C for an additional 4 days . Both control and test birds were sampled using whole carcass rinse techniques and bacteria enumerated using impedance detection times (DT) and total plate count (TPC) techniques at incubation temperatures of 18 and 42 C . The DT and TPC showed a significant increase (P less than .05) in the number of organisms recovered at 18 C but did not show a significant increase (P less than .05) in organisms recovered at 42 C when carcasses were held at 4 C continuously for up to 10 days . Carcasses that were temperature abused at 10 C for up to 24 h were not significantly different (P less than .05) from the unabused controls, using either DT or TPC, following incubation at 18 and 42 C . At an incubation temperature of 18 C, carcasses abused at 15 or 20 C had significantly more growth of bacteria than controls at 1, 6, 12, and 24 h and 4, 8, and 12 h as determined by DT and at 6, 12, and 24 h and 1, 2, 4, 8, and 12 h, respectively, as determined by TPC.(ABSTRACT TRUNCATED AT 250 WORDS) FEMS Microbiol Lett, 1992 Aug 1, 74(1), 27 - 36 Mycobacterium avium and Mycobacterium intracellulare chromatotypes defined by curvilinear gradient HPLC of mycolic acids; Duffey PS et al.; Seventy-nine strains of Mycobacterium avium complex bacteria (MAC), previously characterized by genetic probe analysis, were assayed using two methods of reverse phase high-performance liquid chromatography (HPLC) that employed curvilinear gradients . Although different in column length and cycle time, the methods produced equivalent results, yielding seven distinct chromatographic patterns (chromatotypes) of M . avium and M . intracellulare based on the ratio of mycolate concentrations in the late vs . the middle of three peak clusters (L:M ratio) . The M . avium strains (n = 36) were assigned to chromatotypes 1 through 4 (L:M ratios less than 3), and the M . intracellulare strains (n = 25) to chromatotypes 5 through 7 (L:M ratios greater than 4) . Of 18 Mycobacterium 'X' strains, seven resembled M . avium, seven others resembled M . intracellulare, and four were intermediate between M . avium and M . intracellulare. Mol Pharmacol, 1992 Aug, 42(2), 280 - 5 Anti-liver microsomes autoantibodies and dihydralazine-induced hepatitis: specificity of autoantibodies and inductive capacity of the drug; Bourdi M et al.; Anti-liver microsomes (anti-LM) autoantibodies in patients with dihydralazine-induced hepatitis were found to react specifically with cytochrome P4501A2 (P4501A2) but not with P4501A1 expressed in yeast and bacteria . These results were confirmed by immunoinhibition of methoxyresorufin-O-demethylase activity (supported by the P4501A subfamily); anti-LM antibodies more strongly inhibited this activity in yeast expressing P4501A2 than in yeast expressing P4501A1 . Anti-LM were shown to be specific to the disease; in three cases, these autoantibodies were present at high titers during disease, whereas the titers decreased upon recovery and became undetectable a few months after recovery . Thus, there exists a time-dependent relationship between the disease and the autoantibodies, which does not prove that the autoantibodies are causative of the hepatitis; they might only be a marker . The inductive capacity of dihydralazine toward P450 was also studied . In rats treated in vivo and in human hepatocytes treated in vitro with dihydralazine, a 2-fold increase in P4501A2- and P4501A-supported monooxygenase activities was found . The levels of the other P450 isoforms tested were unchanged during treatment, both in vivo in rats and in vitro in cultures of human hepatocytes . In human hepatocytes, dihydralazine produced a dose-dependent increase in the level of P4501A up to 0.1 mM; induction of P4501A was less strong at 0.2 mM and disappeared at 0.5 mM . The same treatment did not change the level of P4503A4, taken as control . The strong heterogeneity in the expression of P4501A enzymes in human liver and the capacity of these enzymes for induction by dihydralazine and by other compounds might be predisposing factors in this autoimmune disease. Clin Geriatr Med, 1992 Aug, 8(3), 499 - 511 Salivary gland dysfunction; Atkinson JC et al.; Clinically significant declines of salivary output are not a function of aging . Complaints of oral dryness (xerostomia) are common in the elderly patient, however . This complaint is most often a consequence of medications; tricyclic antidepressants, certain antihypertensives, and anticholinergics can cause marked decreases in salivary flow rates . Other medications such as diuretics may cause xerostomia without decreasing salivary output . Systemic diseases such as Sjogren's syndrome can destroy salivary glands . Specific serologic, ophthalmologic, and salivary findings are necessary for the diagnosis of Sjogren's syndrome . Radiation therapy for treatment of head and neck cancers also can damage salivary glands permanently . Infectious agents such as mycobacteria, Epstein-Barr virus, and various oral bacteria can infect human salivary glands . Diminished salivary output can lead to serious oral sequelae . Rapidly progressing dental caries and oral candidiasis are found frequently in this age group . Close supervision of the geriatric patient's oral and dental health is essential. Curr Opin Rheumatol, 1992 Aug, 4(4), 494 - 9 Infections and the immunopathogenesis of seronegative spondyloarthropathies; Keat A; Evidence is accumulating for invasion of joints by bacterial antigens in the seronegative spondyloarthropathies . In spite of the inability to isolate bacteria directly from joint material, indirect evidence, provided by antibody and cellular immune studies, points to a local response to local antigen within the joint . Using monoclonal or polyclonal antibodies, several groups have reported detection of bacterial antigens in synovial fluid cells and synovium itself both in reactive arthritis and in undifferentiated forms of seronegative arthritis . There appear to be direct parallels with animal models insofar as bacterial persistence and arthritogenicity are concerned . To date, however, attempts to detect bacterial DNA in joint material, with the possibility of viability that such findings might imply, have been negative. Am J Med, 1992 Aug, 93(2), 177 - 80 Reactive hemophagocytic syndrome--a clinicopathologic study of 40 patients in an Oriental population; Wong KF et al.; PURPOSE: The purpose of this investigation was to assess the clinical spectrum and outcome of reactive hemophagocytic syndrome (RHS) in an Oriental population . PATIENTS AND METHODS: The investigation was a retrospective study of 40 consecutive patients with RHS diagnosed over a 4-year period in Queen Elizabeth Hospital, a community-based hospital in Hong Kong . RESULTS: The incidence of RHS in Hong Kong appeared to be comparable with that of Western countries . However, the spectrum of diseases associated with RHS was distinctly different . Forty percent of the cases were associated with malignant lymphoma . Another 40% of the cases were associated with infection, often due to bacteria, and infection due to the herpes group of viruses was uncommon, probably reflecting ethnic and geographic differences from the white population . Two patients had the acute lupus hemophagocytic syndrome, a distinctive form of RHS that occurs in patients with systemic lupus erythematosus . Treatment was basically supportive, with specific therapy also given for the underlying disease if present . Eighteen patients (45%) died of the acute disease, mostly as a result of complications of RHS or the underlying diseases . The development of disseminated intravascular coagulopathy was a poor prognostic factor . CONCLUSIONS: RHS is an uncommon disorder in Hong Kong and is often associated with malignant lymphoma or bacterial infection . A rigorous search for an underlying malignancy or infection is thus warranted once this diagnosis is made. Lab Anim Sci, 1992 Aug, 42(4), 352 - 5 Factors associated with intestinal amyloidosis in pigtailed macaques (Macaca nemestrina); Ellsworth L et al.; Factors associated with intestinal amyloidosis in pigtailed macaques (Macaca nemestrina) were studied in 74 cases at the Washington Regional Primate Research Center . The medical records of monkeys during the 5-year period from 1983 to 1988 were analyzed to determine the age at death, age at first episode of diarrhea, number of episodes of diarrhea, episode and cumulative duration of diarrhea, and etiologies of diarrhea . Univariate analysis, using one control for each case, indicated that only episode duration was related to intestinal amyloidosis . Affected monkeys had significantly longer mean episode durations of diarrhea . None of the etiologies examined--bacteria, protozoa, fungi, and simian retrovirus--were significant risk factors for amyloid deposition in the intestinal tract. Nippon Rinsho, 1992 Aug, 50(8), 1724 - 9 {Inflammation and pro-inflammatory cytokine}; Mukaida N; The main pathological feature of inflammation consists of leukocyte infiltration and exudation of plasma into the lesion in the early stage followed by proliferation of connective tissue including fibroblasts, which leads to the formation of granulation tissue . Systemic manifestations include elevation of body temperature and increase of serum acute phase reactants such as C-reactive protein and serum amyloid A . Most of these reactions are presumed to be caused by mediators which are produced by a various kinds of cells and tissues upon contact with inflammatory stimuli, such as bacteria, helminths, viruses, etc . Recently, it has become known that most of inflammatory stimuli induce the production of large amounts of several cytokines including interleukin 1 (IL-1), IL-6, and tumor necrosis factor (TNF) alpha and that these cytokines exert a wide variety of effects to accelerate inflammation . Hence, these cytokines are called "pro-inflammatory cytokines" . These cytokines have overlapping biological functions and induce the production of each other, thus forming a complicated cytokine network . Here, in order to review the roles of these cytokines in the progression of inflammation, the effects on the body temperature, leukocyte infiltration and production of serum acute phase reactants, in particular, will be discussed. Int J Immunopharmacol, 1992 Aug, 14(6), 1037 - 44 Low dose streptozotocin-induced diabetes in mice: reduced IL-2 production and modulation of streptozotocin-induced hyperglycemia by IL-2; Burkart V et al.; The possible role of interleukin 2(IL-2) in the pathogenesis of multiple low dose streptozotocin (Sz)-induced diabetes in mice was analysed . Spleen cells from diabetic male C57Bl/6 mice showed diminished mitogen-induced IL-2 production as determined by bioassay using the IL-2-dependent T-cell line CTLL-2 . In parallel the proliferative response was reduced . Systemic daily administration of human recombinant IL-2 for 3 weeks had dose-dependent effects on the development of hyperglycemia in Sz-treated (5 x 40 mg) mice: while IL-2 at doses of 1 x 2, 1 x 10, 2 x 10 micrograms/kg body weight caused partial suppression of hyperglycemia, higher doses (2 x 20, 2 x 40 micrograms/kg) had an enhancing effect . Treatment with the lowest dose (1 x 1 micrograms/kg) or with a control preparation from bacteria (2 x 10 micrograms/kg) did not significantly alter the course of diabetes . Effects of IL-2 were similar when treatment was started concomitantly with or only after streptozotocin injections . This observation argues against the direct interaction between IL-2 and streptozotocin but suggests modulation of immune reactivity by IL-2 . Our findings of decreased mitogen-stimulated IL-2 production by splenic lymphocytes, and the disease-modulating effect of IL-2 in the low-dose streptozotocin diabetes extend our previous observations in spontaneously diabetic BB rats and further support the notion of an involvement of IL-2 in the control of autoimmune diseases. Biophys J, 1992 Aug, 63(2), 397 - 411 Virtual intermediates in photosynthetic electron transfer; Joseph JS et al.; We explore the possibility of virtual transfer in the primary charge separation of photosynthetic bacteria within the context of several types of experimental data . We show that the peak that might be expected in the virtual rate as electric fields vary the intermediate state energy is severely broadened by coupling to high-frequency modes . The Stark absorption kinetics data are thus consistent with virtual transfer in the primary charge separation . High-frequency coupling also makes the temperature dependence weak over a wide range of parameters . We demonstrate that Stark fluorescence anisotropy data, usually taken as evidence of virtual transfer, can in fact be consistent with two-step transfer . We suggest a two-pulse excitation experiment to quantify the contributions from two-step and virtual transfer . We show that virtual absorption into a charge transfer state can make a substantial contribution to the Stark absorption spectrum in a way that is not related to any derivative of the absorption spectrum. Curr Opin Cell Biol, 1992 Aug, 4(4), 696 - 702 Sodium cotransport proteins; Wright EM et al.; Significant advances have been made in elucidating the structure of Na+ cotransport proteins . Some fifteen of these low-abundance proteins have been cloned, sequenced and functionally expressed . They are members of the 12 membrane-spanning superfamily and they segregate into two groups, the Na+/glucose (SGLT1) and Na+/Cl-/GABA (GAT-1) families . SGLT1 transporters are expressed in bacteria and animal cells, while GAT-1 transporters are mostly expressed in the brain . None have yet been found in plants. J Laryngol Otol, 1992 Aug, 106(8), 695 - 6 Nasal polypectomy: should antral washout be a routine? Dowell M, Pahor AL. Seventy patients with nasal polyps were studied . Forty-seven of these had pre-operative sinus radiographs and all seventy patients had antral washouts at the time of nasal polypectomy . One hundred and seventeen antral aspirates were sent for culture and microscopy . The naked eye appearance of aspirates were turbid in 34 patients . Bacteria were cultured from 15 of the specimens in 10 patients . It was concluded that antral washout should be recommended in all patients who have nasal polypectomy and that there is no need for sinus radiographs in those patients who have uncomplicated nasal polyps. Nippon Sanka Fujinka Gakkai Zasshi, 1992 Aug, 44(8), 987 - 93 {The present and the future of treatment of male infertility}; Kobayashi T; About 90% of male infertile patients are complaint of semen with poor quality, and of which majority are caused by idiopathic disturbance of spermatogenesis . To date, various trials have been made to stimulate spermatogenesis by means of pharmaceutical administrations, their efficacies were, however, poor as expected . The treatment of the patients with poor quality semen is, therefore, mainly focused on artificial insemination, such as intrauterine insemination (IUI) and in vitro fertilization-embryo transfer (IVF-ET), etc . (1) To inseminate the sperm artificially, it is necessary to separate progressively motile sperm with normal morphology from seminal plasma, immotile and abnormal sperm, leucocytes and bacteria . There are two methods for separating progressively motile sperm, one is the density gradient centrifugation and another is the diffusion by their own motility . We have developed various types of density gradients using Percoll, a modified silica gel; the mono-layer Percoll method and the cushion method are employed for sperm concentration, and the discontinuous Percoll density gradient with 4 steps and the continuous-step density gradient are capable of separating progressively motile sperm . The continuous-step density gradient have been employed for 271 cases of IUI, and successful 84 pregnancies were obtained with the pregnancy rate of 30.9% . (2) Cryopreservation of sperm produce various advantages in the treatment of male infertility . Cryoaccumulation of oligozoospermic semen is effective for obtaining a sufficient number of sperm, and frequent insemination with cryopreserved sperm increase the chance of fertilization . To improve the quality of cryopreserved sperm, ejaculated semen was concentrated prior freezing by means of the continuous-step density gradient.(ABSTRACT TRUNCATED AT 250 WORDS) Kansenshogaku Zasshi, 1992 Aug, 66(8), 1120 - 4 {A case of tracho-bronchial aspergillosis complicated with acute myeloblastic leukemia}; Saburi Y et al.; A 59-year-old male was admitted to our hospital in Jan . 1991 with complaints of general malaise and palpitation . Laboratory findings on admission showed anemia, thrombocytopenia and leukopenia consisted of 2.0% myeloblasts with Auerbodies . The bone marrow study showed granuloid hyperplasia with 45.5% myeloblasts . The diagnosis of acute myeloblastic leukemia (M1) was made . After BHAC-AMP therapy, he obtained complete remission . However, he complained of fever and cough, and his chest X ray film showed a focal infiltrative shadow in the right upper lung field . Antibiotics for bacteria and fungus were administered and the abnormal shadow improved in a week . However, as he had hemosputum, the bronchoscopic examination was performed, and multiple ulcers covered by yellow-white tissue were revealed on the wall of the trachea and bilateral main bronchi . Biopsy specimens obtained by transbronchial biopsy showed bronchial aspergillosis . Though intravenous infusion and inhalation of amphotericin B were effective for aspergillosis, he had a relapse of the leukemia and died in autumn, 1991. Kansenshogaku Zasshi, 1992 Aug, 66(8), 1084 - 9 {Detection of Legionella pneumophila using a nested polymerase chain reaction}; Higa F et al.; We evaluated the usefulness of a Nested PCR method for detecting Legionella pneumophila . This method resulted in L . pneumophila specific detection as far as we evaluated . The first and second step PCR achieved the sensitivity as small as 10 pg and 10 fg of the target DNA, respectively . In the detection from Legionella seeded sputa, the method could detect 0.1 cfu/ml of the bacteria, and it took about 12 hours to detect the target DNA . We demonstrated that the Nested PCR method was superior in sensitivity and rapidity for isolation of the bacteria to the conventional using low pH treatment and selective media for Legionella. J Dairy Res, 1992 Aug, 59(3), 265 - 71 Machine-induced teat tissue reactions and infection risk in a dairy herd free from contagious mastitis pathogens; Zecconi A et al.; Machine-induced changes in teat thickness were measured in a randomly selected group of 22 cows from a commercial dairy herd consisting of 110 cows during an experiment lasting 1 year . Half the cows used were free from mastitis whereas the remainder had at least one quarter infected by environmental pathogens . Teats were classified according to a threshold change in teat end thickness of 5% . The relationships between quarter infection or teat duct colonization and teat end thickness changes induced by machine milking were investigated . Teats with greater than 5% change in thickness had significantly increased teat duct colonization and a slight, non-significant, increase in quarter infection . We conclude that machine-induced changes in teat end thickness are predisposing factors for teat duct colonization by environmental pathogens . This may provide an increased new infection risk, especially when hygiene is poor. J Antibiot (Tokyo), 1992 Aug, 45(8), 1207 - 15 New cyclodepsipeptides, enniatins D, E and F produced by Fusarium sp . FO-1305; Tomoda H et al.; New cyclodepsipeptides named enniatins D, E and F were isolated from the culture broth of Fusarium sp . FO-1305 as inhibitors of acyl-CoA:cholesterol acyltransferase (ACAT) . The respective structures of enniatins D, E and F were determined to be cyclo{D-alpha-hydroxyisovaleryl(D-Hiv)-L-N-methylleucinyl(L- Me-Leu)-D-Hiv-L-N-methylvalinyl(L-Me-Val)-D-Hiv-L-Me-Val}, a mixture of cyclo-{D-Hiv-L-Me-Leu-D-Hiv-L-N-methylisoleucinyl(L-Me-Ile)-D-Hiv- L-Me-Val} and cyclo(D-Hiv-L-Me-Ile-D-Hiv-L-Me-Leu-D-Hiv-L-Me-Val), and cyclo(D-Hiv-L-Me-Leu-D-Hiv-L-Me-Ile-D-Hiv-L-Me-Ile) by spectral analyses and chemical degradation . The IC50 values of enniatins D, E and F for ACAT activity in an enzyme assay using rat liver microsomes were calculated to be 87, 57 and 40 microM, respectively. Cryobiology, 1992 Aug, 29(4), 500 - 10 Cryoinjury in human granulocytes and cytoplasts; Yang H et al.; Although most isolated cells can be successfully cryopreserved, human granulocytes have little functional recovery after cryopreservation, even under optimized conditions . Cytoplasts, which are vesicles created from human granulocytes by depletion of organelles including granules and the nucleus, can carry out some of the complex functions of the parent granulocyte such as phagocytosis of bacteria, even after cryopreservation . Human granulocytes and cytoplasts were used in this comparative study of low-temperature responses to assess the relative importance of the plasma membrane and the granules in cryoinjury to human granulocytes . Boyle-van't Hoff plots of cell volume as a function of the reciprocal of osmolality showed that granulocytes and cytoplasts have similar osmometric behavior and equivalent osmotically inactive fractions . The hydraulic conductivities were also similar, indicating that the osmotic properties of the plasma membrane and cytoplasm were retained during preparation of the cytoplasts . Assessment of membrane integrity using fluorescein diacetate after graded freezing stresses showed that the low-temperature responses of cytoplasts were similar to those of human lymphocytes and hamster fibroblasts, with recoveries much higher than those of human granulocytes, particularly after post-thaw incubation at 37 degrees C . The results indicate that the plasma membrane is not the primary site of injury to granulocytes during freezing and thawing, and suggest that activation of cytoplasmic elements, such as granules, may constitute the early events in cryoinjury to human granulocytes . These studies have significance in approaches to the cryopreservation of granulocytes and other types of cells, such as platelets, with increased sensitivity to the conditions encountered during freezing and thawing. Semin Immunol, 1992 Aug, 4(4), 217 - 26 Antigen processing in the mucosal immune system; Keren DF; The mucosal immune system is concerned with host defense along the moist surfaces of the body which have contact with the external environment . These sites contain specialized lymphoid structures which contain precursors for IgA-synthesizing B lymphocytes and immunoregulatory T lymphocytes which will determine whether oral tolerance or a strong immune response develops against antigens administered orally . The key step to antigen processing in the gastrointestinal tract involves its initial uptake from the gut lumen by specialized follicle associated epithelium called 'M' cells . M cells originate from adjacent crypt epithelium and are interspersed between the absorptive epithelial cells in the follicle-associated epithelium . M cells cells have short, irregular microvilli, are closely associated with lymphocytes, do not have a prominent terminal web, and have only weak alkaline phosphatase activity but strong nonspecific esterase activity . M cells do not express surface MHC class II (HLA-DR) antigens . These cells take up macromolecules, viruses, bacteria and protozoa within 30 minutes from the initial presentation of the antigen to the intestinal lumen . After the initial uptake of antigen by M cells, the antigens are transported into the follicular areas to be processed by dendritic cells and brought into close contact with the antigen-specific precursors for IgA secreting plasma cells . The final result of M cell processing is the production of a vigorous secretory IgA response and local cell-mediated immunity with suppression of a systemic IgG, IgE and delayed-type hypersensitivity to orally-administered antigens. J Am Soc Nephrol, 1992 Aug, 3(2), 121 - 7 How tonicity regulates gene expression; Burg MB et al.; The expression of a number of different mammalian genes is directly affected by hypertonicity . At present, the list of their products includes aldose reductase, heat shock proteins, early response factors, and transporters for betaine, inositol, and taurine . Hypertonicity increases the abundance of the mRNAs for all of them . Aldose reductase mRNA levels increase because of increased transcription with little change in the stability of its mRNA . Transcription of the betaine transporter also increases . The mechanisms by which hypertonicity increases the transcription of these mammalian genes remain speculative . However, the consideration of transcriptional control of betaine transport in bacteria and of heat shock proteins in many organisms provides interesting insight into this question. Biol Reprod, 1992 Aug, 47(2), 220 - 6 Human placenta constitutively produces interleukin-8 during pregnancy and enhances its production in intrauterine infection; Shimoya K et al.; Interleukin-8 (IL-8) exerts unique chemotactic and activating activity on neutrophils . To address the significance of IL-8 in the fetoplacental unit during pregnancy, we cultured human placental explants that had been obtained by vaginal delivery, Caesarean section, or artificial abortion and then measured the IL-8 titer in the culture supernatants by enzyme immunoassay (EIA) . Chorionic tissue from the first trimester produced a significant amount of IL-8 (2.2 +/- 0.4 ng/ml/10 mg, n = 5), while placentae in the second trimester (8.3 +/- 1.6 ng/ml/10 mg, n = 7) or at term (9.2 +/- 0.7 ng/ml/10 mg, n = 29) produced significantly higher amounts of IL-8 . The presence or absence of labor did not affect the amount of placental IL-8 production . However, placentae with chorioamnionitis (25.2 +/- 1.6 ng/ml/10 mg, n = 9) showed significantly higher IL-8 production than those without chorioamnionitis (p less than 0.0001) . Northern blot analysis of IL-8 mRNA expression demonstrated a constant level during pregnancy with or without chorioamnionitis, indicating the possibility that the major site of regulation of IL-8 synthesis in the placenta is posttranscriptional . Immunohistochemical analysis of first and third trimester placental tissues with rabbit anti-IL-8 antibody revealed the IL-8 producing cells to be trophoblasts and macrophage-like cells . IL-8 produced by the placental cells might contribute to potentiation of the immunocompetence of placental cells against bacteria invading the fetoplacental unit. Clin Immunol Immunopathol, 1992 Aug, 64(2), 145 - 52 Antigenic determinants of the 70-kDa subunit of the Ku autoantigen; Abu-Elheiga L et al.; Autoantibodies against Ku antigen were found in subsets of sera from patients with rheumatic diseases . The Ku autoantigen was characterized as a DNA-binding protein complex composed of two subunits, 70 and 86 kDa . In this study, we report the amino acid sequences of the 70-kDa subunit that are important for interactions with a monoclonal and autoimmune antibodies . Full-length cDNA and numerous 5' and 3' deletion mutants were expressed in bacteria and the immunoreactivity of the fusion proteins was analyzed by Western blotting . The reactivity of the monoclonal antibody depended on the region between Ile321 and Phe350 . Ten autoimmune sera were tested for reactivity with deletion mutants in immunoblots . The reactivity of six sera strongly depended on the C-terminal amino acids and four sera did not show such dependence; however, these C-terminal sequences did not react with the sera when expressed alone . These results strongly suggest the conformational nature of the Ku autoepitopes . Interestingly, the DNA-binding activity of this Ku protein subunit analyzed by Southwestern blot depended on the same C-terminal amino acids that were involved in interactions with autoantibodies, indicating that anti-Ku autoantibodies are directed to conformationally intact Ku antigen . Reactivities of the autoimmune sera with Met1-Arg115, Met116-Val149, and Val149-Arg586 were also observed . These results demonstrate that different amino acid regions can be involved in interactions with autoimmune antibodies. Eur J Immunol, 1992 Aug, 22(8), 2055 - 62 Virus infection blocks the processing and presentation of exogenous antigen with the major histocompatibility complex class II molecules; Domanico SZ et al.; Helper T cell recognition of antigen requires that antigen be processed and presented by class II expressing antigen-presenting cells (APC) . Many antigens presented by the immune system are part of infectious organisms, for example, bacteria and viruses, which themselves may affect APC function . Here we show that infection of B cell lines as APC with viruses of two different families, namely, influenza A or vaccinia, completely block processing and presentation of an exogenous globular protein antigen pigeon cytochrome c . The block appears to be primarily within the processing pathway, as virus infection has little effect on the presentation of an antigenic peptide of pigeon cytochrome c which does not require processing . It is likely that several steps in the processing pathway are affected . Only live infectious virus, not UV-inactivated virus blocks APC function, indicating that there is no competition of viral particles with cytochrome c for the class II processing machinery . As compared to uninfected cells, virus-infected cells internalize less antigen bound to surface Ig but degrade a similar portion of that which enters the cell . Virus infection results in reduced protein synthesis in APC which may also be a factor in decreasing APC function . Significantly, we show that the processing of a high affinity evolutionary variant of cytochrome c from Drosophila melanogaster is reduced less by virus infection as compared to c . Such knowledge may guide the selection of antigenic epitopes in vaccine design. Trends Biochem Sci, 1992 Aug, 17(8), 295 - 9 The emergence of the chaperone machines; Georgopoulos C; To ensure proper polypeptide folding, oligomerization and transport, elaborate molecular 'chaperone machines' have evolved . These machines are usually composed of a major chaperone protein that binds promiscuously to nascent, unfolded, misfolded or aggregated polypeptides and a set of chaperone 'cohorts', whose function is to enhance efficiency and ensure recycling . These chaperone machines can function by themselves or synergistically to carry out their various tasks. Infect Immun, 1992 Aug, 60(8), 3303 - 8 Role of carbohydrate recognition domains of pertussis toxin in adherence of Bordetella pertussis to human macrophages; van't Wout J et al.; Pertussis toxin (PT) and filamentous hemagglutinin can each mediate the association of Bordetella pertussis with human macrophages . Adherence via filamentous hemagglutinin leads to integrin-mediated entry and survival of the bacteria within the human cell . We determined the contribution of PT to bacterial adherence to human macrophages . Plating macrophages on wells coated with recombinant PT subunit 2 (S2) or S3 decreased PT-dependent bacterial binding by greater than 60%; S1, S4, and S5 were ineffective . S3-dependent adherence was reduced 63% +/- 8% by sialic acid, while S2-dependent adherence was reduced 53% +/- 11% by galactose . Loss of the carbohydrate recognition properties of S2 by deletion of residues 40 to 54 or site-specific mutations at Asn-93, His-47, or Arg-50 eliminated the ability of the subunit protein to competitively inhibit bacterial binding . Peptides corresponding to residues 28 to 45 of S2 and S3 competitively inhibited adherence . Treatment of macrophages with antibodies to Le(a) or Le(x) but not CD14, CD15, CD18, or HLA interfered with PT-mediated binding . Exposure of the macrophages to the B oligomer, S2, or S3 increased binding to the CD11b/CD18 integrin . These results indicate that the carbohydrate recognition domains of both S2 and S3 participate in adherence of B . pertussis to human macrophages . The PT receptor(s), as yet unidentified, appears to carry the Le(a) or Le(x) determinants and is functionally capable of modulating integrin-mediated binding to the macrophage. Arch Oral Biol, 1992 Aug, 37(8), 597 - 601 Susceptibility of Porphyromonas gingivalis and P . asaccharolytica to the non-oxidative killing mechanisms of human neutrophils; Odell EW et al.; Neutrophils are essential for host defence against bacterial dental plaque and the pathogenic bacterial species within it, but in anaerobic environments such as the gingival crevice neutrophils can kill bacteria only with non-oxidative microbicidal compounds stored in their granules . Porphyromonas gingivalis W83, a pathogenic plaque species, and the avirulent non-oral type-strain P . asaccharolytica were incubated anaerobically with intact neutrophils and with compounds extracted from normal human neutrophil granules . The killing of bacteria and the inactivation of lysozyme, cathepsin G, elastase, bacterial-permeability increasing factor and defensins by culture supernatants were assayed . P . asaccharolytica but not P . gingivalis was killed under anaerobic conditions by intact neutrophils . P . gingivalis was also resistant to neutrophil granule compounds, its viability being reduced from a mean of 3.3 x 10(6) to 6.1 x 10(4) c.f.u/ml in 60 min by 400 micrograms/ml neutrophil granule extract, as compared to a reduction from 4.4 x 10(6) to 2.3 x 10(3) c.f.u/ml for P . asaccharolytica . P . gingivalis culture supernatant inactivated cathepsin G, elastase, bacterial-permeability increasing factor and defensins . Resistance to neutrophil non-oxidative killing mechanisms may be an important virulence factor for P . gingivalis. Br J Cancer Suppl, 1992 Aug, 18, S8 - 12 The aetiology of cancer in the very young; Buckley JD; Epidemiological studies of cancer in young children have implicated a number of environmental factors, which need to be studied in more detail, but it is probably fair to say that the main benefit of these studies has come from the negative findings, which have served to exclude (or at least place an upper limit on the role of) potential risk factors . Our inability to identify environmental causes could mean either that the environment does not substantially affect cancer incidence in young children, or that we are simply not looking in the right places . Most attention has naturally been focused on the known and suspected environmental carcinogens and mutagens . Based on the data summarised in this paper, one possibility is that the most important mutagen is endogenous: 5-methyl-cytosine . If so, factors increasing cancer risk could be those which increase the rate of spontaneous deamination, or impair the efficiency of the excision repair enzymes, or regulate the processes of CpG methylation and demethylation . These factors could still be classed as mutagens in their own right, since they would lead to an increase in uncorrected point mutations, but they would be distinctive in a number of ways . Firstly, since the role of methylation in bacteria is very different from that in eukaryotic cells the Ames mutagenicity assay could not be relied upon to detect methylation-mediated mutagens . Furthermore the risk may be highly age dependent, reflecting changes in methylation patterns with growth and cellular differentiation . Agents which disrupted the imprinting process in the testis would not be detectable by animal carcinogenicity tests unless specifically looked for.(ABSTRACT TRUNCATED AT 250 WORDS) Infect Immun, 1992 Aug, 60(8), 3231 - 7 Differential morphologic and metabolic alterations in permissive versus nonpermissive murine macrophages infected with Legionella pneumophila; Yamamoto Y et al.; Legionella pneumophila infection of macrophages from permissive guinea pigs and from A/J mice compared with infection of cells from nonpermissive BDF1 mice was studied by electron microscopy . The cells from the BDF1 mice were nonpermissive for legionella growth in vitro and showed few if any bacteria in phagosomes by electron microscopic examination . Similar electron micrographic examination of macrophages from A/J mice permissive for legionella growth showed numerous intact intracellular bacteria within 24 to 48 h of culture and the transition of intracellular bacteria from localization in a few large vacuoles early in the course of infection to later localization in areas surrounded and studded by ribosomes . These electron microscopic observations were similar to those seen in the case of guinea pig macrophages infected with legionellae . Biochemical studies of macrophages from permissive versus nonpermissive animals showed little or no differences in respiratory burst and lysosomal enzyme activity for macrophages from all animals tested . However, when zymosan was used as a stimulant, macrophages from the nonpermissive mouse strain produced a larger amount of H2O2 and O2- than did cells from permissive guinea pigs or A/J mice . However, legionella vaccine itself induced no detectable or very little H2O2 and O2- in macrophages tested from any source . These results suggest that permissiveness of A/J mouse macrophages to legionella growth may involve mechanisms similar to those occurring in guinea pig macrophages in terms of morphologic and possibly even biochemical events . The relatively higher production of reactive oxygens by BDF1 mouse macrophages in response to zymosan correlated with nonpermissiveness for legionella growth, although further analysis is necessary to link these observations. Carbohydr Res, 1992 Jul 20, 232(1), 89 - 105 Synthesis of 4'-deoxy-4'-fluorokanamycin A and B; Takahashi Y et al.; 4'-Deoxy-4'-fluorokanamycins A (17) and B (25) have been prepared through fluorinative ring-opening of the D-galacto-3',4'-oxiranes (8 and 21) derived from kanamycin A and B with potassium hydrogenfluoride in ethane-1,2-diol . The mechanism of preponderant formation of the 4'-deoxy-4'-fluoro-D-gluco (9 and 22) over the 3'-deoxy-3'-fluoro-D-gulo derivatives was discussed . In the synthesis of 25, the unusual 3',6'-epimine (23) was the main product along with the 4'-deoxy-4'-fluoro derivative . The mechanism of this reaction is also discussed . Both 17 and 25 were active against resistant bacteria producing aminoglycoside-adenylylating enzymes for HO-4'. Experientia, 1992 Jul 15, 48(7), 650 - 6 Heat shock proteins in autoimmune disease . From causative antigen to specific therapy? Yang XD, Feige U. Heat shock proteins (hsp) are highly conserved from bacteria to man . Bacterial hsp, with approximate molecular weights of 60 kDa (hsp60), are immunodominant antigens that are immunologically cross-reactive with their mammalian counterparts . Hsp molecules are therefore useful in studies of fundamental questions concerning immune responses to foreign as opposed to self antigens . The finding that immune responses to hsp are associated with both experimentally-induced and spontaneous autoimmune diseases in animals has prompted intensive research to assess the role of bacterial hsp as the etiological agents involved in the development of autoimmune diseases . Recent evidence from animal models of autoimmune disease has clearly demonstrated the involvement of hsp in both the pathogenesis and the immunoregulation of autoimmune diseases . Studies with arthritogenic and diabetogenic T cell clones have identified immunogenic epitopes of hsp . These have been shown to ameliorate adjuvant arthritis in Lewis rats, and insulin-dependent diabetes mellitus (IDDM) in non-obese diabetic (NOD) mice . Such studies may have important therapeutic implications for the future treatment of human autoimmune disease. Proc Natl Acad Sci U S A, 1992 Jul 15, 89(14), 6521 - 5 Modulation of adenylate cyclase toxin production as Bordetella pertussis enters human macrophages; Masure HR; During the course of human infection, Bordetella pertussis colonizes sequential niches in the respiratory tract that include intracellular and extracellular environments . In vitro the expression of virulence factors such as the adenylate cyclase toxin is coordinately regulated by the bvg locus, which is an example of a two-component sensory transduction system . With this toxin as a reporter, enzyme activities were compared between a wild-type and an altered strain to determine whether bacterial entry into human macrophages affected gene expression . BPRU140, a strain containing an inducible expression vector, produced enzyme activity independent of bvg . Samples of the parent, the induced, and the uninduced BPRU140 were incubated individually with macrophages for 30 min . Extracellular bacteria were then killed by gentamicin . The number of viable intracellular bacteria and the internalized bacterial enzyme activity were measured over time . By 2.5 hr all samples reached a steady-state concentration of 10(5) bacteria per 10(6) macrophages . Following an initial peak of enzyme activity, adenylate cyclase values for the parent and the uninduced BPRU140 decreased to a basal level, while the values for the induced strain remained at least 3-fold greater . Therefore, compared with the persistence of enzyme in the induced strain BPRU140, the decrease in enzyme production by the parent and the uninduced BPRU140 upon entry into macrophages indicates in vivo down-modulation of gene expression . These observations support the hypothesis that sensory transduction contributes to adaptations for bacterial survival in the infected host. Biochim Biophys Acta, 1992 Jul 15, 1131(3), 325 - 8 The primary structures of the flavodoxins from two strains of Desulfovibrio gigas . Cloning and nucleotide sequence of the structural genes; Helms LR et al.; The structural genes coding for the flavodoxin proteins from two different strains of the sulfate-reducing bacteria Desulfovibrio gigas (ATCC 19364/NCIB 9332 and ATCC 29494/DSM 496) have been identified, cloned and the nucleotide sequence established . The protein sequences derived from the gene from each strain share a sequence identity of 66% with regions directly involved in binding the flavin mononucleotide cofactor being the most homologous . Both aromatic residues that flank the flavin isoalloxazine ring in the crystal structure of the flavodoxin from D . vulgaris, i.e., Trp-60 and Tyr-98, are also present in these flavodoxin proteins . These observations stand in contrast to reports that the flavodoxin from Desulfovibrio gigas contains a single tryptophan residue which is located distant from the flavin binding site . Therefore, the FMN binding site of this flavodoxin is not distinct from the other Desulfovibrio flavodoxins in this regard. Mech Ageing Dev, 1992 Jul 15, 64(3), 303 - 13 Age-related changes in neutrophil structure and function; Perskin MH et al.; The elderly suffer higher rates of morbidity and mortality from infectious disease than younger adults . Since neutrophils are the first line of defense against infection the vulnerability to infection of the elderly may be due, at least in part, to age-related changes in neutrophils (PMNs) . Previous studies have suggested that there is an age-related increase in plasma membrane microviscosity in lymphocytes and neurons of the elderly . We have explored the hypothesis that there is a similar change in plasma membrane viscosity in the neutrophil and that this alteration in the neutrophil plasma membrane leads to diminished neutrophil function . When we studied plasma membrane viscosity of neutrophils by determining the fluorescence anisotropy of 1-(4-trimethylaminophenyl)-6-diphenyl-1,3,5-hexatriene labeled cells, we were surprised to find that plasma membrane viscosity is decreased in the neutrophils of the elderly as compared to young controls (microviscosity parameter, 1.33 +/- 0.07, in the elderly, n = 21, versus 1.62 +/- 0.06, in young controls, n = 19, P less than 0.004) . As expected, stimulated (FMLP 0.1 microM) O2- generation by neutrophils from the elderly was significantly decreased compared with young controls (34 +/- 7% decrease, n = 6, P less than 0.04) . Both resting and stimulated neutrophils demonstrated an age-related decline in adherence to a component of the extracellular matrix, denatured collagen (gelatin, P less than 0.04, n = 22 elderly subjects) . In contrast, neutrophils from the elderly adhered to fibrinogen at least as well as neutrophils from young controls . Moreover, chemotaxis to activated complement components (in zymosan activated plasma) and FMLP did not change with increased age whether studied in the Boyden chamber (92 +/- 7% control, n = 14) or under agarose (90 +/- 13% control, n = 11) . These studies suggest that an age-related decrease in plasma membrane viscosity is associated with a decrease in O2- production and adherence of neutrophils to components of the extracellular matrix . Thus, age-related alterations in neutrophil structure may result in diminished neutrophil function and increased susceptibility to infection with pyogenic bacteria. Science, 1992 Jul 10, 257(5067), 241 - 5 Peptide binding by chaperone SecB: implications for recognition of nonnative structure; Randall LL; The molecular basis for recognition of nonnative proteins by the molecular chaperone SecB was investigated with an in vitro assay based on the protection of SecB from proteolysis when a ligand is bound . The SecB tetramer has multiple binding sites for positively charged peptides . When the peptide binding sites are occupied, the complex undergoes a conformational change to expose hydrophobic sites that bind the fluorescent probe 1-anilinonaphthalene-8-sulfonate . A model is proposed for interaction of nonnative polypeptides with both hydrophilic and hydrophobic sites on SecB. Regul Pept, 1992 Jul 2, 40(1), 87 - 100 Purification of cationic cystine-rich peptides from rat bone marrow . Primary structures and biological activity of the rat corticostatin family of peptides; Belcourt D et al.; Seven cationic, cystine-rich peptides of 29 to 32 amino acid residues have been purified from extracts of rat bone marrow (R-1, R-1a, R-1b, R-2, R-3, R-4 and R-5) . Structural analysis clearly indicated that all seven peptides belong to the corticostatin/defensin family of leukocyte-derived peptides known to participate in oxygen-independent killing of phagocytosed bacteria . For R-1 to R-5, six cysteine residues were found at characteristic and highly conserved positions . R-1a and R-1b were partially characterized and appear to be structural variants of R-1 . Aside from the conserved cysteines, there is a remarkable degree of structural diversity evident within the sequences of those members of the corticostatin/defensin family characterized so far . The structures of the peptides that we have purified can be compared directly with the sequences obtained for rat defensins isolated from extracts of peritoneal neutrophils (Lehrer, Ganz and Selsted, Cell, 64 (1991) 229-230) . Some discrepancies are apparent which can be explained in terms of proteolytic cleavage of several of these peptides at both amino- and carboxyl-termini . The corticostatins owe their bioactivity to their ability to compete with corticotropin for occupancy of the corticotropin receptor (Zhu, Hu, Mulay, Esch, Shimasaki and Solomon, Proc . Natl . Acad . Sci . USA, 85 (1988) 592-596) . The potency of these peptides can be expressed in terms of their capacity to inhibit the steroidogenic response of isolated rat adrenocrotical cells half-maximally stimulated by corticotropin (i.e., at the ED50 concentration for corticotropin in this assay, namely 33 pM) . In this assay, the rat peptides R-1, R-2 and R-3 were shown to be inactive . In contrast, the more cationic peptides R-4 and R-5 were found to inhibit steroidogenesis . R-4 was somewhat less active than rabbit corticostatin (IC50 25 nM) showing an IC50 value of 50 nM . R-5 appeared to be significantly less potent than R-4 . The lower yield of R-5 precluded an accurate estimate of the corticostatic potency of this peptide . R-4 differs in structure from R-5 in having an arginine to serine substitution at position 7 . It can be concluded that an arginine at this position accounts, at least in part, for the corticostatic activity of R-4.(ABSTRACT TRUNCATED AT 400 WORDS) Diagn Microbiol Infect Dis, 1992 Jul, 15(5), 483 - 7 The E-Test as an epidemiologic tool; Baquero F et al.; A comparison was made of the discriminating power of the E-Test and conventional methods to distinguish different bacterial strains harboring plasmid-mediated extended-spectrum beta-lactamases . The E-Test provided quantitative data comparable to those of a broad range of concentrations in the agar dilution method, but proved to be far superior to the disk diffusion test or the broth microdilution semiautomatic PASCO system for the detection and hence for the epidemiologic surveillance of this type of resistant strain. Prostaglandins, 1992 Jul, 44(1), 75 - 86 Eicosanoid synthesis and Helicobacter pylori associated gastritis: increase in leukotriene C4 generation associated with H . pylori colonization; Ahmed A et al.; The importance of pro-inflammatory leukotriene C4 in Helicobacter pylori (H . pylori) associated gastritis in man is unknown . Fresh gastric biopsy specimens from 28 dyspeptic patients were obtained: 10 showed normal antral histology with no evidence of H . pylori, the remaining 18 patients exhibited histological gastritis and were H . pylori positive as assessed by histology, culture and urease test . Twelve of these 18 patients received 240 mg twice daily colloidal bismuth subcitrate for four weeks before re-endoscopy . Gastric biopsies from H . pylori positive patients were incubated under basal and Ca(2+)-ionophore mediated conditions: Radioimmunoassay analysis of the supernatant showed basal release of prostaglandin E2 and leukotriene C4 was slightly but not significantly elevated in H . pylori positive mucosa . However in H . pylori positive mucosa there was an 85% increase in leukotriene C4 synthesis when biopsies were incubated with ionophore, compared to only 13% increase in H . pylori negative mucosa (p less than 0.02) . After eradication of H . pylori by colloidal bismuth subcitrate, there was a clearance of inflammatory cell infiltrate as assessed by histology and a significant reduction in ionophore-mediated leukotriene C4 formation compared with before treatment (p less than 0.02) . These results suggest that H . pylori gastritis is associated with increased capacity to generate leukotriene C4, which may amplify the damaging effects of the bacteria on gastric mucosa. Genomics, 1992 Jul, 13(3), 672 - 80 A yeast artificial chromosome contig encompassing the type 1 neurofibromatosis gene; Marchuk DA et al.; The yeast artificial chromosome (YAC) system (Burke et al., 1987, Science 236: 806-812) allows the direct cloning of large regions of the genome . A YAC contig map of approximately 700 kb encompassing the region surrounding the type 1 neurofibromatosis (NF1) locus on 17q11.2 has been constructed . A single YAC containing the entire NF1 locus has been constructed by homologous recombination in yeast . In the process of contig construction a novel method of YAC end rescue has been developed by YAC circularization in yeast and plasmid rescue in bacteria . YACs containing homology to the NF1 region but mapping to another chromosome have also been discovered . Sequences of portions of the homologous locus indicate that this other locus is a nonprocessed pseudogene. Genetics, 1992 Jul, 131(3), 753 - 60 Inference of horizontal genetic transfer from molecular data: an approach using the bootstrap; Lawrence JG et al.; Inconsistencies in taxonomic relationships implicit in different sets of nucleic acid sequences potentially result from horizontal transfer of genetic material between genomes . A nonparametric method is proposed to determine whether such inconsistencies are statistically significant . A similarity coefficient is calculated from ranked pairwise identities and evaluated against a distribution of similarity coefficients generated from resampled data . Subsequent analyses of partial data sets, obtained by the elimination of individual taxa, identify particular taxa to which the significance may be attributed, and can sometimes help in distinguishing horizontal genetic transfer from inconsistencies due to convergent evolution or variation in evolutionary rate . The method was successfully applied to data sets that were not found to be significantly different with existing methods that use comparisons of phylogenetic trees . The new statistical framework is also applicable to the inference of horizontal transfer from restriction fragment length polymorphism distributions and protein sequences. Clin Exp Immunol, 1992 Jul, 89(1), 154 - 7 Functionally active complement is present in human ovarian follicular fluid and can be activated by seminal plasma; Perricone R et al.; Human ovarian preovulatory follicular fluids (FF) from 10 women were analysed for their complement contents . Functionally active complement was detected in all the fluids studied in amounts similar to those present in normal human serum . Pooled FF was challenged by seminal plasma in order to determine whether seminal plasma could activate FF complement, the pattern of such an activation and the possible consequences on the reproductive function . FF complement activation occurred during the incubation with seminal plasma with features including alternative pathway activation, factor B and C3 conversion and reduction in total haemolytic complement, as well as an inhibition by seminal plasma of the FF complement response to a new activating challenge . Possible consequences for fertilization, implantation of a fertilized ovum and local defence mechanisms against viruses and bacteria are discussed. Mol Cell Biol, 1992 Jul, 12(7), 3006 - 14 Fused protein domains inhibit DNA binding by LexA; Golemis EA et al.; Many studies of transcription activation employ fusions of activation domains to DNA binding domains derived from the bacterial repressor LexA and the yeast activator GAL4 . Such studies often implicitly assume that DNA binding by the chimeric proteins is equivalent to that of the protein donating the DNA binding moiety . To directly investigate this issue, we compared operator binding by a series of LexA-derivative proteins to operator binding by native LexA, by using both in vivo and in vitro assays . We show that operator binding by many proteins such as LexA-Myc, LexA-Fos, and LexA-Bicoid is severely impaired, while binding of other LexA-derivative proteins, such as those that carry bacterially encoded acidic sequences ("acid blobs"), is not . Our results also show that DNA binding by LexA derivatives that contain the LexA carboxy-terminal dimerization domain (amino acids 88 to 202) is considerably stronger than binding by fusions that lack it and that heterologous dimerization motifs cannot substitute for the LexA88-202 function . These results suggest the need to reevaluate some previous studies of activation that employed LexA derivatives and modifications to recent experimental approaches that use LexA and GAL4 derivatives to detect and study protein-protein interactions. Immunology, 1992 Jul, 76(3), 355 - 61 Distinctive western blot antibody patterns induced by infection of mice with individual strains of the Mycobacterium avium complex; Elsaghier A et al.; Systemic infection of mice with organisms of the Mycobacterium avium complex (MAC) induced antibody responses, characteristic for each of the three tested individual strains . The influence of host genetic factors was reflected up to 3 months after infection by the finding of generally oligobanded and multibanded Western blot patterns in C57B1/6 and BALB/c mice, respectively . Nevertheless, more bands developed at 6 months in C57BL/6 mice . The response to three antigens of 18,000, 38,000 and 24,000 MW was analysed in greater detail . Antibodies to a protease-resistant 18,000 MW band produced only by BALB/c mice were either strain specific, following infection with M . avium, strain Maa-B2, or cross-reactive within MAC, following infection with M . avium strain Maa-A6 and M . paratuberculosis, strain Map-203 . Another protease-resistant antigen of 38,000 MW was immunogenic only in Maa-B2 infected mice . This constituent was found to be related to the protease-sensitive antigen of corresponding molecular weight from M . tuberculosis . Two 24,000 MW proteins of M . paratuberculosis were separated by two-dimensional gel electrophoresis: antibodies to the anodic band were induced by Map-203 infection, whilst the cathodic band was revealed by heteroclitic antibodies from Maa-B2-infected mice . The latter antigen is apparently expressed during in vivo replication, but not during in vitro culture of Maa-B2 bacteria . We generally conclude, that the selective antibody patterns after live infection, could be attributed to differences in the release of native antigens within mycobacterial lesions . In view of a high degree of species specificity, some of the immunogenic constituents identified may also be useful for serodiagnostic application. J Infect, 1992 Jul, 25(1), 83 - 7 Pericarditis, pneumonia and brain abscess due to a combined Actinomyces--Actinobacillus actinomycetemcomitans infection; Zijlstra EE et al.; A patient is described in whom there developed a pulmonary infiltrate and constrictive pericarditis caused by a combined Actinomyces and Actinobacillus actinomycetemcomitans infection, presumably originating from his poor dentition . The diagnosis was only made following repeated thoracotomy . After surgery, long-term treatment with antibiotics led to complete clinical recovery . None the less, some months later he was found to have a brain abscess which resolved during a further course of antibiotics . The variable clinical picture of actinomycosis is discussed, as well as the role of other bacteria frequently associated with actinomycotic infection, in particular Actinobacillus actinomycetemcomitans . The therapeutic implications are described. FEMS Microbiol Lett, 1992 Jul 1, 73(1-2), 95 - 9 Relationship between electroporation conditions, electropermeability and respiratory activity for Frankia strain ACN14a; Cournoyer B et al.; The use of electroporation for introducing macromolecules into intact cells of the actinomycete Frankia was investigated . Electropermeability was demonstrated by the uptake of dextran (70 kDa) molecules labeled with fluorescein isothiocyanate (FITC) inside Frankia cells . Upon pulsation with an exponentially decaying electric field, the cell membranes became permeable . Loading increased with initial pulsed electric field strength and capacitance . Increased loading efficiency was inversely related to INT (2-(p-iodophenyl-3-(p-nitrophenyl)-5- phenyltetrazolium chloride) reduction activity (respiring bacteria) of the cell population . The presence of CaCl2 in the electroporation and resealing buffer raised INT-reduction activity but K2SO4 decreased this activity . Resealing of electropores was confirmed by a decreasing FITC-dextran loading through the recovery period . The use of FITC-dextran molecules and INT-reduction assay are two new approaches for the study of permeabilization and cellular activity of electroporated bacteria. FEMS Microbiol Lett, 1992 Jul 1, 73(1-2), 105 - 10 The detection of two antigenic groups among Renibacterium salmoninarum isolates; Bandin I et al.; The analysis of the membrane proteins and their antigenic properties in a group of 14 geographically diverse strains of Renibacterium salmoninarum revealed the existence of antigenic diversity within this species . Eleven isolates, including the type strain ATCC 33209, shared a similar protein profile with a major component of 57 kDa whereas three strains showed a common pattern with a major protein of 30 kDa . The quantitative agglutination tests and Western blotting assays seem to indicate the existence of serological heterogeneity, with two distinct groups being detected. Arch Dis Child, 1992 Jul, 67(7 Spec No), 808 - 12 Receiver operating characteristic curves for comparison of serial neutrophil band forms and C reactive protein in neonates at risk of infection; Russell GA et al.; The performance of indirect indices of infection in the newborn vary because of differences in techniques, including diagnostic cut off levels . We have compared serial neutrophil band cell counts with C reactive protein measured by rate nephelometry . The 'gold standard' was a positive culture and the performance of the tests was compared by the technique of receiver operating characteristics (ROC) as well as sensitivity and specificity . A total of 172 septic screens were performed in 56 patients . The operational diagnostic cut off values were: C reactive protein greater than 8 mg/l, immature:total neutrophil ratio (I:T ratio) greater than 0.2, and band count greater than 5% . Compared with the sensitivity of C reactive protein (71%), I:T ratio (34%) was significantly different but band count (69%) was not . The specificity of C reactive protein (72%) was better than band count (39%) but no better than I:T ratio (73%) . ROC curves were constructed for all possible diagnostic cut off values of the tests and superior performance was demonstrated for C reactive protein compared with band count and I:T ratio . We conclude that C reactive protein is a useful early indicator of infection in neonates and that ROC curves permit comprehensive and graphic comparison between tests and the calculation of optimal diagnostic cut off values. Arch Dis Child, 1992 Jul, 67(7), 952 - 5 Occurrence of suppurative lymphadenitis after a change of BCG vaccine; Hengster P et al.; BCG vaccines are the oldest vaccines in use today, but the protective effect of the vaccination is still controversial . The risk of contracting tuberculosis is low compared with the possible complications after this vaccination . In Austria the formerly used BCG vaccine was not available in the required amount and another vaccine was released by the drug authorities . This product, with a more virulent strain, was used between August and December 1990, and this increased the incidence of complications . Eighty four of 1950 vaccinated newborn babies developed severe suppurative lymphadenitis three to 28 weeks after the vaccination, and surgical treatment was found to be necessary . Isoniazid treatment did not prove to be successful when the lymph node exceeded a certain size . Culture was successful in 46% up to week 20; after 20 weeks no culture became positive . All cultured bacteria were isoniazid sensitive . The question of general vaccination is raised and several points were considered before we came to the conclusion that except for high risk groups a general vaccination programme for neonates is not justified in Western countries. J Mol Evol, 1992 Jul, 35(1), 51 - 9 Unusual organizational features of the Drosophila Gart locus are not conserved within Diptera; Clark DV et al.; The Drosophila Gart locus consists of two genes . One gene encodes three enzymes in the de novo purine nucleotide biosynthesis pathway {glycinamide ribonucleotide synthetase (GARS), aminoimidazole ribonucleotide synthetase (AIRS), and glycinamide ribonucleotide transformylase (GART)} . The second gene lies within an intron of the purine gene and encodes a cuticle protein . To investigate the evolution of the Gart locus, the Chironomus tentans homolog was cloned by screening a genomic DNA library with a polymerase chain reaction product . This study shows that the interesting structural features of this locus conserved in two distant Drosophila species are not found in the Chironomus homolog . These features include the cuticle protein gene nested within an intron and the existence of an alternative transcript to yield a monofunctional enzyme . In addition, the extremely rapid divergence of coding sequence seen for members of the tandemly duplicated AIRS domain in Drosophila is found to be much less rapid in Chironomus. Handchir Mikrochir Plast Chir, 1992 Jul, 24(4), 210 - 4 {Mycobacterium malmoense infection of the flexor tendon sheath}; Osterwalder C et al.; Infections of the tendon sheath caused by non-tuberculous mycobacteria are well known . So far, only one case involving Mycobacterium malmoense has been reported . The authors report of a 73-year old patient having undergone carpal tunnel release . In spite of granulomatous inflammation of the flexor tendon sheaths, there was no biopsy cultured for acid-fast bacteria . One year later, symptoms recurred and synovialectomy was repeated . In spite of macroscopic appearance of the synovialis and histological studies, a specific diagnosis was not yet made . Two weeks later, acid-fast bacteria, identified as M . malmoense were cultured in the BACTEC vial . Because specific treatment was not initiated, a further recurrence developed five months later, necessitating further surgery with synovialectomy . Properly treated with Rifampin and Ethambutal for twelve months, the patient has remained asymptomatic for the following two years. FEMS Microbiol Immunol, 1992 Jul, 4(5), 273 - 9 Intracellular growth of Mycobacterium avium in human macrophages is linked to the increased synthesis of prostaglandin E2 and inhibition of the phagosome-lysosome fusions; Rastogi N et al.; A virulent strain of Mycobacterium avium grew actively inside human adherent peripheral blood monocyte-derived macrophages . Bacteria were always confined to the phagosome compartment and were encapsulated . Cytochemical labeling of acid phosphatase using transmission electron microscopy showed a strong inhibition of the phagosome-lysosome fusions (PLF) in macrophages as not more than 25-30% bacteria containing phagosome at any time effectively fused with lysosomes . In case of a positive fusion event, the bacterial capsule prevented the diffusion of the lysosomal contents to the bacterial surface . Moreover, the infection of macrophages both by living and gamma-killed M . avium was linked to an increased synthesis of prostaglandin E2 (PGE2); however the total amount of PGE2 synthesized in the latter case was significantly lower than that observed with viable organisms . Our results suggest that the inability of human macrophages to control M . avium infection is linked to immunosuppressive pathways, e.g . enhanced synthesis of PGE2 and also to an impairment of normal microbicidal functions of the infected macrophages. J Gen Microbiol, 1992 Jul, 138 ( Pt 7), 1503 - 13 Identification of erythrocyte-binding antigens in Helicobacter pylori; Huang J et al.; The surface antigens of Helicobacter pylori conferring erythrocyte-binding activity were obtained by adsorption onto formaldehyde-treated dog and goat erythrocytes from supernatant fractions of sonicated bacteria and elution using a high concentration of NaCl . The desorbed material was analysed by SDS-PAGE and immunoblotting with anti-whole-cell serum to agar-grown bacteria which had been absorbed with broth-grown, non-haemagglutinating cells (haemagglutination-associated antiserum) . Two polypeptides with molecular masses of 25 and 59 kDa were revealed as erythrocyte-binding antigens . Strains which agglutinated both dog and goat erythrocytes possessed both these erythrocyte-binding antigens, whereas an antigenically cross-reactive 24 kDa polypeptide was present in a strain which only agglutinated goat erythrocytes . Haemagglutinin material was extracted from H . pylori using n-octylglucopyranoside and purified by Sepharose chromatography and sucrose density gradient ultracentrifugation . The purified extract directly agglutinated erythrocytes in a neuraminyl-lactose-sensitive and neuraminidase-sensitive manner . The 59 kDa polypeptide was not present in the purified haemagglutinin preparation . The haemagglutination-associated antiserum reacted strongly with the 25 kDa polypeptide band which was the most prominent polypeptide band on analysis of the purified haemagglutinin preparation by SDS-PAGE and silver staining . Thus, H . pylori possesses at least two adhesins, one of which recognises a N-acetylneuraminic acid (alpha 2-3) moiety of receptors, the other being of unknown receptor specificity . Differences in the antigenicity and molecular masses of these adhesins in individual strains may underlie differences in receptor-binding specificities and haemagglutination profiles. J Gen Microbiol, 1992 Jul, 138 ( Pt 7), 1479 - 87 The use of rRNA sequences and fluorescent probes to investigate the phylogenetic positions of the anaerobic ciliate Metopus palaeformis and its archaeobacterial endosymbiont; Embley TM et al.; The polymerase chain reaction (PCR) was used to amplify small-subunit ribosomal DNA from the anaerobic ciliated protozoon Metopus palaeformis, and from its uncultured endosymbiotic bacteria . This was accomplished directly from total DNA extracted from protozoa without prior isolation or enrichment for symbiont cells . The double-stranded amplification products were precipitated and directly sequenced using the linear PCR reaction . Fluorescent oligonucleotide probes were designed and used in whole-cell hybridizations to provide direct visual evidence that the sequences originated from the host ciliate and from the endosymbiont . Phylogenetic analysis of the Metopus palaeformis sequence consistently placed it as a deep-branching lineage near the root of the ciliate tree . However, the present data were insufficient to resolve the detailed relationship between Blepharisma and Metopus and thus to determine if the heterotrichs are mono- or paraphyletic . Phylogenetic analysis of the symbiont partial sequence clearly demonstrated that it is an archaeobacterium and that it is closely related to, but distinct from, Methanobacterium formicicum. J Periodontol, 1992 Jul, 63(7), 567 - 75 Effect of a social cognitive intervention on oral health status, behavior reports, and cognitions; Tedesco LA et al.; An intervention designed to test the influence of cognitive restructuring on protective oral health behaviors was conducted with 108 patients with mild to moderate gingivitis . Subjects in the experimental group viewed slides of active, mobile bacteria taken from their mouths on 5 occasions: before and after prophylaxis and at 3 appointments, one month apart . A specially trained hygienist discussed with these participants the process of periodontal disease, the role of bacteria, and self-efficacy (self-control) for oral hygiene self-care . Both experimental and control group subjects received instruction in oral self-care procedures . Assessments of oral health using Loe and Silness' plaque and gingival indices (PI and GI) were taken throughout the study and at 3- and 6-month follow-up visits . Self-efficacy, oral hygiene intentions, attitudes, and values comprised the set of cognition variables . Plaque and gingival indices mean differences between groups approached significance at visit 6 . Analyses were also performed using percent of gingival surfaces scored at "0" (no visible bleeding on probing) . A trend occurred for group differences in percent "0" scores at visit 6, with the experimental group maintaining higher percent zeros (better health) at this 3-month follow-up . At visit 7 (9-month follow-up), PI and GI differences disappeared . No significant differences were found between groups for oral health cognitions or behavior reports over time . The data suggest that the cognitive-behavioral intervention produced a delayed relapse in protective oral self-care behaviors, and by extension, oral health status . Such a delay could be clinically relevant in promoting adherence to oral hygiene behavior between professional visits. Am J Vet Res, 1992 Jul, 53(7), 1172 - 5 Cytologic findings of tracheobronchial aspirates from 66 thoroughbred racehorses; Sweeney CR et al.; Tracheobronchial aspirates obtained from 66 healthy Thoroughbred racehorses in training at the same track were examined . Twenty-seven percent of the horses had greater than 20% neutrophils in the aspirate . Eosinophils, mast cells, giant cells, and Curschmann's spirals of mucus were observed in 94, 83, 65, and 42% of the horses, respectively . Hemosiderophages were observed in 86% of the horses, half of which had previous confirmation of exercise-induced pulmonary hemorrhage . Although fungal elements were seen in 70% of the horses, bacteria were detected in only 3% of the horses . The authors conclude that inflammatory airway disease is widespread in the racing Thoroughbred population. J Parenter Sci Technol, 1992 Jul-Aug, 46(4), 102 - 6 The Water Pressure Integrity Test--a new integrity test for hydrophobic membrane filters; Dosmar M et al.; Sterilizing grade hydrophobic filters are used for the sterile filtration of gases in pharmaceutical and biological applications . Until now the integrity of these membrane filters and their ability to retain bacteria, has been correlated to a solvent based nondestructive integrity test . Current methods use solvents to wet the membranes in order to perform bubble point and diffusion integrity tests . Solvent based integrity tests make it difficult to test in situ following sterilization because of the risk of downstream solvent contamination . A newly developed method, the Water Pressure Integrity Test (WPIT), allows for the integrity testing of hydrophobic filters eliminating the problems associated with traditional test methods employing solvents . A prime advantage of WPIT is that it may be performed in situ post sterilization without any downstream manipulations . The test has been directly correlated to the retention of bacterial challenges . Data will be provided to show the reliability and sensitivity of this easy to perform test. Am J Obstet Gynecol, 1992 Jul, 167(1), 139 - 43 Randomized, double-blind study of cefotetan and cefoxitin in post-cesarean section endometritis; MacGregor RR et al.; OBJECTIVE: The hypothesis of the study is that cefotetan and cefoxitin will be equally efficacious and safe in the treatment of post-cesarean section endometritis . STUDY DESIGN: In a double-blind, randomized manner 140 patients with post-cesarean section endometritis were treated with cefotetan, 2 gm intravenously every 12 hours, or cefoxitin, 2 gm intravenously every 6 hours . They were followed prospectively for clinical response and side effects . Cure rates between the two groups were compared with the chi 2 test . RESULTS: The cure rates were 83% for cefotetan and 79% for cefoxitin (p = 0.56) . No patient required a change in therapy due to adverse effects, and no abnormal bleeding occurred . CONCLUSION: In this study cefotetan and cefoxitin appeared equally effective in treating endometritis with no difference in side effects or complications. Trans R Soc Trop Med Hyg, 1992 Jul-Aug, 86(4), 399 - 400 Pneumocystis carinii as a cause of pneumonia in HIV-infected patients in Lusaka, Zambia; Machiels G et al.; PIP: 4 cases of Pneumocystis carinii pneumonia in HIV-infected patients studied at the University of Zambia Medical School, Lusaka, were verified by bronchoalveolar lavage . Pneumocystis is common in North American AIDS patients, but has been considered rare in Africa . One reason may be that facilities for diagnosis, bronchoscopy with bronchoalveolar lavage, are not usually available . 44 consecutive HIV seropositive patients who were unresponsive to a 10-day course of antibiotics, and whose sputum was negative for acid fast bacteria, underwent bronchoalveolar lavage from February 1990 to December 1990 . HIV status was assayed with Welcozyme ELISA kits, and P . carinii was detected with toluidine blue O stain . The 1st case of confirmed P . carinii pneumonia was a 35-year old man who had a productive cough for 4 weeks, fever, and dyspnea . He was treated with co-trimoxazole and was symptom-free in 3 weeks, but developed severe Stevens-Johnson reaction . His cultures were positive for M . tuberculosis at week 8 . He was lost to follow-up . The 2nd case was a 26-year old man with a 6-month history of cough and white sputum, treated without effect with antituberculous medication . He improved over 3 weeks with co-trimoxazole, but died of respiratory failure 2 months later . The 3rd case was a 30-year old woman being treated for pulmonary tuberculosis, who became progressively dyspneic 7 months later . She developed a generalized maculo-papular rash after taking co-trimoxazole, so was given dapsone 100 mg/day, prednisone 1 mg/kg/day, and trimethoprim 15 mg/kg for 1 week . She improve in 3 weeks . The 4th case was a 30-year old man with a 4-week history of dry cough and dyspnea and recent high fever . He was given co-trimoxazole, but developed generalized purpura after 5 days . His treatment was changed to Dapsone 100 mg/day, prednisone 1 mg/kg/day, and antituberculous medication . He improved after 3 weeks, and is being maintained on Fansidar 1 tablet/week . These cases are remarkable because 2 of them also had pulmonary tuberculosis, which is often the presumed diagnosis of pneumonia in African AIDS patients . Furthermore, 3 developed serious drug reactions to co-trimoxazole, also considered an uncommon occurrence . Mol Biol (Mosk), 1992 Jul-Aug, 26(4), 859 - 68 {Localization of conserved and variable segments in amino acid sequences of homologous proteins using a personal computer}; Kostetskii PV et al.; A set of aligned homologous protein sequences is divided into two groups consisting of the most related sequences m and k . The value of the position variability of homologous protein sequences is defined as a number of failures to coincide in the intergroup comparison of all possible k x m pairs of amino acid residues in that position divided by k x m . The position variability value plotted vs the sequence position number with a window of 10 positions gives the intergroup local variability profile . The area S of the figure included between the local variability profile and the straight line corresponding to the mean local variability value is compared with the average area S(r) for 1000 random homologous protein families . If S is greater than S(r) by more than 2 standard deviation units sigma r the local variability profile is assumed to contain peaks and hollows corresponding to significant variable and conservative regions of the sequences . The profile extrema containing the area surplus delta S = S-(S(r) + 2 sigma r) are cut off by two straight lines to locate significant regions . The numerical experiment on the family of homologous phospholipases A2 revealed the linear dependence of the values S(r) and sigma r upon the position variability standard deviation sigma v of the homologous sequences . Furthermore, it was shown for protein families of various length (rhodopsins, aspartate aminotransferases, cytochromes b, L- and M-subunits of photosynthetic bacteria photoreaction centre and alpha-subunits of Na, K-ATPase), that delta S = S - n(S'r + 2 sigma r), where S - the area of the local variability profile, n = L/l (L - the length of the given protein family and l - the length of the hypothetical protein domain) . If l = 250 then S'r = -1.42 + 62.56 sigma v and sigma'r = -0.14 + 7.46 sigma v. J Am Podiatr Med Assoc, 1992 Jul, 82(7), 388 - 91 Management of complications associated with an excision of a lipoma from the ankle; McIver W et al.; This case study demonstrates the following: close proximity of lipoma to underlined skin can compromise circulation; the size of the lipoma and the resulting redundant skin after the incision can lead to fluid development and impede the healing process; and a patient who is noncompliant and stands or walks excessively after the excision of a lipoma can cause buildup of a hematoma, which can compromise the incision site . Lipomas associated with the foot or ankle are rarely symptomatic . When they do cause pain and are surgically treated, possible complications because of their size and location must be considered . Finally, these complications could have been avoided if the redundant skin had been excised and a Penrose drain inserted to eliminate excess fluid buildup . Also, better communication should have been maintained with the patient during the entire postoperative course. J Biochem (Tokyo), 1992 Jul, 112(1), 40 - 4 Synthetic reaction of Cellvibrio gilvus cellobiose phosphorylase; Kitaoka M et al.; The synthetic reactions of the cellobiose phosphorylase from Cellvibrio gilvus were investigated in detail . It was found that, besides D-glucose, some sugars having substitution or deletion of the hydroxyl group at C2 or C6 of the D-glucose molecule could serve as a glucosyl acceptor, though less effectively than D-glucose . The enzyme showed higher activity with beta-D-glucose than with the alpha-anomer as an acceptor . This result indicates that it recognizes the anomeric hydroxyl group not involved directly in the reaction . beta-D-Cellobiose was also phosphorolyzed faster than the alpha-anomer . Substrate inhibition was observed with D-glucose, 6-deoxy-D-glucose, or D-glucosamine as an acceptor, with D-glucose being most inhibiting . This inhibition was studied in detail and it was found that D-glucose competes with alpha-D-glucose-1-phosphate for its binding site . A model of competitive substrate inhibition was proposed, and the experimental data fit well to the theoretical values that were calculated in accordance with this model. J Protozool, 1992 Jul-Aug, 39(4), 521 - 5 The interface between air and water: a perturbation source eliciting adaptive behaviour in ciliates; Ricci N et al.; Interference with the water-air interface, both direct (by contact with a flat, rigid surface) and indirect (by inducing a meniscus) caused the ciliated protozoa we investigated to actively collect in the water column or on the substrate directly under the area of altered surface tension . A crowding effect is observed in this "rest area" reaching plateau values within one hour after onset of the experiment . The simple experimental procedures described here induced analogous behaviour in both Paramecium caudatum (a swimmer) and Oxytricha bifaria (a crawler) . The ciliates seem in this reaction to be seeking a refuge from vibrations transmitted by the free interface . Our discovery is discussed in its implications for the adaptive biology and ecology of these micro-organisms. J Dent Assoc S Afr, 1992 Jul, 47(7), 333 - 6 Plaque quantitation through protein measurement; Smit A et al.; This study was undertaken to establish whether the quantitation of dental plaque protein by a dye-binding method (Coomassie G-250) may be used as an index of the amount of dental plaque sampled . Ten sites were sampled in 34 children on 5 occasions at 4 month intervals . The mean protein concentration in 1391 plaque samples was 6.9 +/- 4.1 micrograms (micrograms) (mean +/- standard deviation) . A three-way analysis of variance showed that the plaque protein concentration was similar at the different sampling sites in the same child (p = 0.14), but statistically significant differences were observed with respect to time of sampling (F = 36.24; p = 0.0001) and individual sampled (F = 5.69; p = 0.0001) . These observations indicate that plaque bacterial counts may be expressed as units of protein concentration and this method may be useful to relate the number of viable bacteria to an estimate of the amount of plaque collected . This ratio allows standardisation for any variation in the amount of plaque collected. Transpl Int, 1992 Jul, 5(3), 133 - 8 Pancreatic juice cytology for monitoring pancreatic grafts in the early postoperative period; Kubota K et al.; Thirty-one pancreas transplant recipients were monitored by pancreatic juice cytology in the early postoperative period . An increase in the total amount of cells and, in particular, signs of immunoactivation with the appearance of two or more blast-transformed cells per specimen were taken as evidence of acute rejection . According to these criteria a total of 38 rejection episodes were diagnosed . The first positive cytology appeared after 9 days (mean) and lasted for 2 days (mean) . Immunocytochemical analysis of the juice showed increased amounts of CD3+ cells during rejection . When rejection occurred during prophylaxis with antithymocyte globulin, neutrophils were preponderant in the pancreatic juice while during OKT-3 prophylaxis a high percentage of monocytes was a characteristic finding . Antirejection treatment was started when the cytology became positive and all rejection episodes except one were reversed . A decrease in the pancreatic juice amylase activity occurred in 66% of the rejection episodes, but in only 5 of the 38 episodes was the decrease highly significant . No correlation was found between graft rejection and volume excretion of pancreatic juice . There were no persistent or characteristic changes in serum amylase or peripheral white blood cell count at the time of rejection . Graft pancreatitis was diagnosed cytologically in 7 patients, in 5 of whom the grafts were eventually lost. Infect Immun, 1992 Jul, 60(7), 2718 - 25 Immunological characterization of the lipooligosaccharide B band of Bordetella pertussis; Martin D et al.; Two structurally and immunologically different components of Bordetella pertussis endotoxin can be visualized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining: a major A band and a faster-migrating minor B band . Certain mutant strains of B . pertussis express only the B band, while the wild-type strains produce both lipooligosaccharides (LOS) . Two monoclonal antibodies (MAbs) directed against the minor LOS B band were generated, allowing the study of this surface molecule on different strains of Bordetella . These two MAbs, designated BL-8 and BL-9, reacted strongly with phenol-water-purified LOS obtained from a B . pertussis LOS B mutant strain . Sodium periodate treatment of the purified LOS prevented binding of the MAbs, indicating the carbohydrate nature of the epitope(s) . Western immunoblotting experiments revealed that the epitope(s) recognized by these MAbs is conserved on all B . pertussis and Bordetella bronchiseptica Vir- (avirulent) variant strains tested but is not present on Bordetella parapertussis and B . bronchiseptica Vir+ (virulent) wild-type strains . Further studies showed that although present in the lipopolysaccharide B band expressed by Vir- strains, the epitope(s) recognized by the MAbs is not accessible on the surface of intact B . bronchiseptica cells . For B . pertussis, the density and accessibility of this epitope(s) are dependent on the virulence-associated or LOS phenotype expressed by the strain . Our data demonstrate that the expression and accessibility of the epitope(s) are significantly greater on the LOS B variant strains and LOS AB Vir- strains compared with fresh B . pertussis clinical isolates . For these latter strains, which are Vir+, this epitope(s) was barely detectable on the surface of intact bacteria, despite Western blot analyses that revealed specific reactions between the MAbs and the LOS B band . The two LOS B-specific MAbs had no bacteriolytic activity against a LOS AB wild-type strain, while the control MAb BL-2, which is specific for the B . pertussis LOS A band, significantly reduced the number of living bacteria in the same assay . Moderate lytic activity against a mutant strain expressing only the LOS B band was observed for MAb BL-8 but not for MAb BL-9 or BL-2 . These data demonstrate that the type, amount, and surface exposure of the LOS are related to the phenotype expressed by a specific B . pertussis strain . In addition, the LOS B MAbs also reveal the antigenic conservation of carbohydrate epitopes among B . pertussis and B . bronchiseptica strains. Int J Immunopharmacol, 1992 Jul, 14(5), 761 - 6 Upregulation of adhesion molecules induced by broncho-vaxom on phagocytic cells; Duchow J et al.; Whole blood was incubated with the bacterial extract Broncho-Vaxom (OM85) at various concentrations and for different periods of time . Expression of the beta 2-integrins (LFA-1, CD11a/CD18; MAC-1, CD11b/CD18; p150,95, CD11c/CD18) and ICAM-1 (CD54) by monocytes and granulocytes was studied using flow cytometry . OM85 enhanced the expression of MAC-1 and ICAM-1 on monocytes and granulocytes in a dose-dependent manner . Maximal expression was achieved with 1 mg/ml bacterial extract . The effect on MAC-1 expression was not due to the low concentration of endotoxin contaminating the preparation (less than 1 ng/mg) since polymyxin-B did not substantially affect the adhesion molecule upregulation induced by OM85 . In addition, OM85 enhanced the expression of p150,95 on monocytes and granulocytes, and also increased expression of LFA-1 on monocytes, but not on granulocytes . While MAC-1 and p150,95 expression reached peak values between 1 and 6 h, levels of ICAM-1 rose constantly for 10 h . We suggest that the clinical interest of OM85 in the management of recurrent infections could be related to be upregulation of adhesion molecules induced by this bacterial extract. J Hosp Infect, 1992 Jul, 21(3), 223 - 9 A randomized study on the effect of bladder irrigation with povidone-iodine before removal of an indwelling catheter; Schneeberger PM et al.; We have evaluated the effect of povidone-iodine (PVP-I) bladder irrigation prior to catheter removal on subsequent bacteriuria . Of 264 patients, 138 received PVP-I irrigation and 126 were controls . Both groups were similar with respect to duration of catheterization and bacteriuria before removal of the catheter . Of 497 cultures taken after catheter removal 99 (20%) were positive . This included 52 of 233 in the control group (22%) and 47 of 264 in the study group (18%) . Patients with positive cultures had a significantly longer period of catheterization . Our trial did not demonstrate that PVP-I bladder irrigation before catheter removal reduces subsequent bacteriuria. J Clin Microbiol, 1992 Jul, 30(7), 1772 - 7 Insertion element IS1081-associated restriction fragment length polymorphisms in Mycobacterium tuberculosis complex species: a reliable tool for recognizing Mycobacterium bovis BCG; van Soolingen D et al.; Recently, the insertion element IS1081 from Mycobacterium bovis was identified . In this study, the usefulness of IS1081 in the epidemiology of tuberculosis was investigated . The host range of this insertion sequence was found to be restricted exclusively to the group of Mycobacterium tuberculosis complex bacteria, whereas none of the 10 mycobacterial species which do not belong to the M . tuberculosis complex contained IS1081-homologous DNA . All 99 M . tuberculosis complex strains investigated carried five or six copies of IS1081, and very limited IS1081-associated restriction fragment length polymorphisms were observed among the strains . Seven different IS1081-containing bands were distinguished in each strain, and the patterns differed only in one or two insertion sequence-containing bands . The banding pattern of M . bovis BCG differed in the presence of a 8.0-kb IS1081-containing PvuII fragment which was absent from all other M . tuberculosis complex strains. J Heart Lung Transplant, 1992 Jul-Aug, 11(4 Pt 2), S232 - 6 Distinguishing between infection, rejection, and the adult respiratory distress syndrome after human lung transplantation; Paradis IL et al.; The adult respiratory distress syndrome, bacterial pneumonia, cytomegalovirus pneumonitis, acute rejection, or a combination thereof were the primary causes of radiographic infiltrates or gas exchange abnormalities that occurred early after lung transplantation . The time of occurrence after transplantation, standard measures of clinical assessment as for nontransplant patients (i.e., vital signs, weight, white blood cell count, sputum, and cultures, etc.), bronchoalveolar lavage, and transbronchial lung biopsy were the primary tools used to analyze these situations . Bacterial pneumonia always occurred after postoperative day 2, acute rejection after postoperative day 5, and cytomegalovirus pneumonitis after postoperative day 16 . Although cultures of bronchoalveolar lavage fluid were useful to detect pneumonia caused by bacteria, virus, and fungus, the types of cells recovered by bronchoalveolar lavage were not diagnostic of any type of disorder . Transbronchial lung biopsy was necessary to detect acute rejection and cytomegalovirus pneumonitis . Thus the cause of an early radiographic infiltrate or impairment of gas exchange was almost always reliably determined by using standard tools of clinical assessment, knowledge of the usual temporal sequence of the complications, and judicious use of bronchoalveolar lavage and transbronchial lung biopsy. J Periodontol, 1992 Jul, 63(7), 611 - 7 Development of a rapid latex agglutination test for periodontal pathogens; Nisengard RJ et al.; The studies reported here describe the development, characterization, and initial application of latex agglutination assays for periodontal pathogens . Latex reagents were prepared by sensitization of latex microspheres with rabbit IgG antibodies to either Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, or Prevotella intermedia . The protein concentration utilized for sensitization and microsphere size were optimized . One reagent was prepared to A . actinomycetemcomitans and a second combination reagent was prepared by mixing latex sensitized with antibodies to P . gingivalis and latex sensitized with antibodies to P . intermedia . The sensitivity of both latex reagents in the traditional wet and a dried format was evaluated . In addition, sensitivity and specificity with homologous and heterologous bacterial suspensions were evaluated . The reagents were found to demonstrate both specificity and sensitivity . Initial studies with subgingival human plaque demonstrated the ability of these reagents to detect the specific organisms in plaque. J Gen Virol, 1992 Jul, 73 ( Pt 7), 1679 - 85 Simian virus 40 (SV40) large T antigen-dependent amplification of an Epstein-Barr virus-SV40 hybrid shuttle vector integrated into the human HeLa cell genome; Stary A et al.; We analysed the DNA rearrangements that occurred during the integration and amplification of an Epstein-Barr virus (EBV)-simian virus 40 (SV40) hybrid shuttle vector in human cells . The human HeLa cell line was episomally transformed with the EBV-SV40 p205-GTI plasmid . After a 2 month culture in a selective medium, a HeLa cell-derived population (H-G1 cells) was obtained in which the p205-GTI vector was integrated as a single intact copy deleted in the EBV latent origin of replication (OriP) . Sequencing data showed that the endpoints of the plasmid sequences, at the plasmid-cell DNA junctions, are located within the two essential elements of EBV OriP, which may form several secondary structures . This result suggests that a specific DNA sequence (OriP) or palindromic structures could play a role in this integration process . This represents the first fully characterized site of integration of an EBV vector in human cells . The transient expression of the SV40 large T antigen in H-G1 cells leads to the appearance of episomal molecules with an extremely heterogeneous size pattern . Individual analysis of these episomes after rescue in bacteria indicated that they retained sequences of both the p205-GTI plasmid and cellular DNA . Comparison of the structure of these circular DNAs with those of the integrated p205-GTI copy indicated that large T antigen expression in human cells leads to the amplification of the integrated shuttle vector according to the 'onion skin' model developed for transformed rodent cells . Indeed, amplified sequences were colinear with the integrated p205-GTI copy and its surrounding cellular sequences, distributed almost equally around the SV40 replication origin, and circularized by illegitimate recombination which did not involve specific nucleotide sequences . This system is of interest in that it enables easy recovery of individual recombined molecules in host bacteria . Each isolated clone contains a unique recombination junction which is easily and rapidly characterized and sequenced. Cancer Res, 1992 Jul 1, 52(13), 3534 - 8 Antitumor activity of magainin analogues against human lung cancer cell lines; Ohsaki Y et al.; Magainin 1 and magainin 2, originally isolated from African clawed frog Xenopus laevis skin, inhibit the growth of bacteria and fungi . Synthetic magainin A (MAG A) and magainin G (MAG G) are more potent against bacteria and protozoa . In order to determine the antitumor activity of these analogues, we have tested these two analogues against six small cell lung cancer (SCLC) cell lines NCI-H82, NCI-H526, NCI-H678, NCI-H735, NCI-H841, and NCI-H889, which were known to differ by more than 10-fold in their sensitivity to different chemotherapeutic agents, and four normal human fibroblast cell lines . Semiautomated 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays of the six SCLC cell lines revealed average concentrations producing 50% inhibition (IC50) values of 2.6 microM (range, 0.49-9.30 microM) for cisplatin, 2.5 microM (range, 0.39-6.00 microM) for etoposide, and 138.8 nM (range, 55.0-450.0 nM) for doxorubicin . The average IC50 of MAG A was 8.64 microM (range, 6.23-11.7 microM) and that of MAG G was 8.82 microM (range, 4.44-12.5 microM) against the SCLC cell lines . Despite a 10-fold difference in sensitivity to standard chemotherapeutic agents, the IC50 of MAG A and MAG G differs by less than 3-fold . The average IC50 against four normal human fibroblast cell lines was 21.1 microM (range, 12.7-25.6 microM) for MAG A and 29.2 microM (range, 21.3-34.8 microM) for MAG G . Combined exposure to the IC50 concentration of MAG A or MAG G plus IC50 of etoposide or cisplatin decreased the percentage of surviving SCLC cells to 29.0% (range, 26.1-31.7%) . MAG A or MAG G had an additive effect when used with standard chemotherapeutic agents . These data suggest that MAG A and MAG G have in vitro antitumor activity against SCLC cell lines. Rev Argent Microbiol, 1992 Jul-Dec, 24(3-4), 171 - 86 {Analysis of principal coordinates and factorial correspondence analysis applied to the taxonomy of type strains of Desulfovibrio}; Pucci OH; Bacterial taxonomy using mathematical methods can be carried out with different techniques . Two techniques are used in this paper: analysis of principal coordinates and factor analysis of correspondences . The first one allows 2 and 3 dimension graphs of bacteria, thus showing their relationship considering proximity . The second one gives new data because it is an analysis which allows to connect bacteria to the reactions which identify them . In order to carry out these analyses 14 strains of the Desulfovibrio genus were studied, thus obtaining a classification consistent with other numeric method (group analysis with Simple Matching coefficient of average bond) to which it adds new information. Schweiz Rundsch Med Prax, 1992 Jun 23, 81(26), 863 - 5 {Current insights on the pathogenesis of chronic inflammatory bowel diseases}; Volk BA et al.; The pathogenesis and etiology of inflammatory bowel disease (IBD) is poorly understood . As a matter of fact, it is not even certain whether either one is a single entity with different forms of clinical manifestations, or whether each one represents a single clearly separable entity . Common features of both diseases are chronic persistence, recurrent exacerbation and remission, the production of autoantibodies, as well as the expression of aberrant HLA-class II molecules on the surface of epithelial cells on the site of inflammation gut . It is likely that these events involve a disturbed immunoregulatory function or autoimmune process . Since the beginning of investigation the cause of IBD, infectious agents (bacteria, virus, mycobacterium paratuberculosis and others) or bacterial products (endotoxin, peptidoglycans from the bacterial cell wall) have been considered as primary causes . Epidemiological studies showed a marked increase of the incidence rates of IBD in industrial countries leading to the hypothesis, that environmental factors could play a role in the pathogenesis of the disease . So far it is clear that the major identified risk factor for IBD is a genetic susceptibility confirmed by studies showing a positive family history. Vet Microbiol, 1992 Jun 15, 31(4), 351 - 62 Serotype specificity of immunological assays for the capsular polymer of Actinobacillus pleuropneumoniae serotypes 1 and 9; Inzana TJ et al.; The cross-reactivity of the purified polysaccharides of Actinobacillus pleuropneumoniae serotypes 1 and 9 were examined using a variety of highly sensitive assays, such as radioimmunoassay, latex agglutination, enzyme-linked immunosorbent assay (ELISA), and immunoblotting . In addition, conventional immunodiffusion was included for comparison . Latex agglutination, utilizing affinity-purified IgG to capsule, was also used to serotype whole cells . Agglutination or precipitation tests (radioimmunoassay, latex agglutination, and immunodiffusion) indicated no cross-reactivity between the capsules of serotypes 1 and 9, and no cross-reactivity between whole cells by latex agglutination . Assays that required binding of the capsule to a solid support (ELISA and immunoblotting) did demonstrate cross-reactions between serotypes 1 and 9 capsules, although reactions with the heterologous serotype were weaker than with the homologous serotype . The cross-reactivity could not be attributed solely to nonspecific factors because similar cross-reactivity did not occur with serotype 5 or 7 capsules by any assay . Reactivity of antisera with homologous or heterologous capsule was reduced, but not completely eliminated, by adsorption with washed, live bacteria of the heterologous serotype . Thus, the assay, as well as the antigen or specificity of the antibody reagent used, may influence the results of A . pleuropneumoniae serotyping or serological tests. J Mol Biol, 1992 Jun 5, 225(3), 891 - 6 Mutation of asparagine 111 of rubisco from Rhodospirillum rubrum alters the carboxylase/oxygenase specificity; Chene P et al.; The conserved asparagine 111 of ribulose-1,5-bisphosphate carboxylase/oxygenase from the photosynthetic bacteria Rhodospirillum rubrum was identified as a candidate for a side-chain that might be involved in the carboxylase/oxygenase specificity . It was replaced by site-directed mutagenesis with aspartic acid, leucine, glutamine or glycine residues . The mutant enzymes exhibit a very low carboxylase activity compared with the wild-type enzyme . The values of Km(RuBP) and kcat for Asn111----Gly, the most active mutant, are 420 microM and 0.034 s-1, compared with 13 microM and 3.0 s-1 for wild-type . The mutation of Asn111----Gly causes a more than tenfold decrease in the CO2/O2 specificity factor, tau, tau Asn111----Gly = 0.56 and tau wild-type = 6.7 . This is the first reported change in rubisco specificity by a single site-directed mutation alone and suggests a target for future protein engineering studies. J Biol Chem, 1992 Jun 5, 267(16), 11483 - 9 A novel mechanism for achieving transgene persistence in vivo after somatic gene transfer into hepatocytes; Wilson JM et al.; Infusion of hepatocyte-specific DNA-protein complexes into rats leads to transient recombinant gene expression in liver . The eventual deterioration of gene expression is due in part to instability of the targeted DNA . In a previous report, we noted retention of transgene sequences in liver and persistent recombinant gene expression when the animals were subjected to partial hepatectomy following in vivo gene transfer . In an attempt to define the mechanism(s) responsible for persistent gene expression following partial hepatectomy, we characterized the molecular state of the retained, liver-associated transgenes . Southern blot analysis of DNA from liver tissues harvested various times after in vivo gene transfer and partial hepatectomy (10 min to 11 weeks) demonstrated high levels of transgene DNA (100-10,000 copies/cell) . The predominant form of this DNA appeared to be episomal based on analyses of uncut DNA or DNA restricted by an endonuclease with one site in the plasmid . Livers from several animals contained a small proportion of transgene sequences of unknown structure . The existence of episomal DNA in liver was confirmed in experiments in which intact plasmid was rescued from total hepatocyte DNA by transformation of bacteria . Both strands of DNA in the liver-associated plasmid retained a bacterial pattern of methylation suggesting that the plasmid had not replicated in the eukaryotic cell . These results are consistent with the hypothesis that the majority of transgene sequences are retained as stabilized plasmids . The specific form of DNA which is transcriptionally active was not identified in these studies . This represents a new mechanism for retaining foreign DNA in eukaryotic cells in vivo and has implications both for the development of somatic gene therapies and the pathogenesis of viral diseases. Carbohydr Res, 1992 Jun 4, 230(1), 89 - 105 Study on fluorination of 2,3-dideoxy-2,3-(N-tosylepimino)-alpha-D-allopyranosides, and synthesis of 3'-deoxy-3'-fluorokanamycin B and 3',4'-dideoxy-3'-fluorokanamycin B; Kobayashi Y et al.; Reaction of the structurally rigid methyl 2,3-dideoxy-4,6-O-isopropylidene-2,3-(N-tosylepimino)-alpha-D-a llopyranoside (6) with KHF2 in DMF at 150 degrees gave initially methyl 2,3-dideoxy-2-fluoro-4,6-O-isopropylidene-3-tosylamido-alpha-D-altrop yranoside (10) by N-tosylepimine-ring opening, and 10 was gradually converted into the stable methyl 2,3-dideoxy-3-fluoro-4,6-O-isopropylidene-2-tosylamido-alpha-D-glucopyra noside (11) . A reversible mechanism involving 6 and 10 has been proposed . In the mobile methyl 2,3-dideoxy-2,3-(N-tosylepimino)-alpha-D-allopyranoside (7) and the corresponding 4,6-di-O-acetyl (8) and -di-O-methyl derivatives (9), reactions with KHF2 proceeded comparatively rapidly giving the corresponding 3-deoxy-3-fluoro-alpha-D-glucopyranosides as the major products . A slightly different reaction mechanism for the mobile compounds has been proposed . By application of this study, 3'-deoxy-3'-fluorokanamycin B was prepared by treatment of 4",6"-O-cyclohexylidene-2'-deamino-3'-deoxy-3'-epi-6'-N-methoxycarbonyl- 1,3, 3"-tri-N-tosyl-2',3'-(N-tosylepimino)kanamycin B (21) with KHF2 as the key reaction . 3',4'-Dideoxy-3'-fluorokanamycin B was also prepared . Both compounds were active against resistant