Biochem J, 1990 Mar 15, 266(3), 785 - 91 Calditol tetraether lipids of the archaebacterium Sulfolobus solfataricus . Biosynthetic studies; Nicolaus B et al.; Lipids from the archaebacterium Sulfolobus solfataricus are based on 72-membered macrocyclic tetraethers made up from two C40 diol units differently cyclized and either two glycerol moieties or one glycerol moiety and a unique branched-chain nonitol named calditol (glycerodialkylnonitol tetraethers, GDNTs) . To elucidate the biosynthesis of calditol and related tetraethers, labelled precursors, {U-14C,1(3)-3H}glycerol, {U-14C,2-3H}glycerol, D-{1-14C,6-3H}glucose, D-{6-14C,1-3H}glucose, D-{1-14C,2-3H}glucose, D-{1-14C,6-3H}fructose and D-{1-14C}galactose, were fed to S . solfataricus . Without regard to stereochemistry or phosphorylation, incorporation experiments provided evidence that the biosynthesis of calditol occurs via an aldolic condensation between dihydroxyacetone and fructose, through a 2-oxo derivative of calditol as an intermediate . The latter is in turn reduced and then alkylated to yield the GDNTs . The biogenetic origins of both glycerol and C40 isoprenoid moieties of GDNTs are also discussed. Exp Neurol, 1990 Mar, 107(3), 208 - 13 Cryopreservation of human brain tissue; Robbins RJ et al.; Tissues from products of conception were examined to determine the feasibility of obtaining viable neural tissue after suction abortion at 9-12 weeks of gestation . The ventral mesencephalon, a prototype region whose maturation can be monitored and which is a potential tissue for transplantation, was identified in 32 of 120 cases . The tissue was then screened for the presence of infectious agents, while being held at -196 degrees C in cryopreservative solutions . Three of 32 specimens were found to be contaminated by normal vaginal bacteria; all other viral, fungal, and mycoplasma testing was negative . Thawed brain fragments retained high viability after storage in liquid nitrogen and when grown in vitro exhibited neuronal morphology, tyrosine hydroxylase immunoreactivity, and dopamine production . We have demonstrated that human fetal brain tissue can be cryopreserved in a manner which not only retains viability but allows normal phenotypic differentiation after thawing. Rev Prat, 1990 Mar 11, 40(8), 709 - 14 {Tuberculosis and mycobacterioses in the AIDS era}; Chretien J et al.; The emergence of the HIV infection has been concomitant with a halt, noticed as early as 1986, in the regression of tuberculosis . The resurgence of tuberculosis, and indeed of all mycobacterioses, is determined by the HIV-induced alteration of cell-mediated immunity, which encourages the development of tuberculous and non-tuberculous bacteria . These bacteria themselves have an immunosuppressant effect on CD4 lymphocytes and macrophages, thus contributing to the progress of the HIV infection . The HIV and tuberculous infections therefore constitute a kind of "diabolical duo", the degree of endemia of one of these two diseases being influenced by the other and reciprocally . The final result is a menace to all countries where tuberculosis is strongly endemic and where the HIV infection is spreading . Only one of these infections, tuberculosis, is within the reach of our therapeutic armentarium . Patients must be investigated for tuberculosis systematically and actively--even though this disease may be hidden by misleading clinical signs and symptoms of AIDS--in order to treat it and separate, at least partially, the diabolical duettists. Nucleic Acids Res, 1990 Mar 11, 18(5), 1133 - 6 Circular chromosomal DNA in the sulfur-dependent archaebacterium Sulfolobus acidocaldarius; Yamagishi A et al.; The shape of the chromosomal DNA of the sulfur-dependent archaebacterium Sulfolobus acidocaldarius was analyzed by the pulsed-field gel electrophoresis(PFGE) . S.acidocaldarius DNA digested with Notl showed two DNA bands at around 1.0 Mbp and 2.1 Mbp . Notl-linking clones were isolated from the library of S.acidocaldarius chromosomal DNA . It contained two Notl sites . Both 1.0 and 2.1 Mbp DNA band separated by PFGE were hybridized with the two independent Notl-linking fragment . Each right and left arms of two Notl-linking fragments were hybridized with one of the two DNA bands separated by PFGE . The results indicated that the chromosomal DNA of S.acidocaldarius is circular. J Biol Chem, 1990 Mar 5, 265(7), 4058 - 63 Purification of the glutamyl-tRNA reductase from Chlamydomonas reinhardtii involved in delta-aminolevulinic acid formation during chlorophyll biosynthesis; Chen MW et al.; The formation of delta-aminolevulinic acid, the first committed precursor in porphyrin biosynthesis, occurs in certain bacteria and in the chloroplasts of plants and algae in a three-step, tRNA-dependent transformation of glutamate . Glutamyl-tRNA reductase, the second enzyme of this pathway, reduces the activated carboxyl group of glutamyl-tRNA (Glu-tRNA) in the presence of NADPH and releases glutamate 1-semialdehyde (GSA) . We have purified Glu-tRNA reductase from Chlamydomonas reinhardtii by employing six different chromatographic separations . The apparent molecular mass of the protein when analyzed under both denaturing (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) and nondenaturing conditions (rate zonal sedimentation on glycerol gradients) was 130,000 Da; this indicates that the active enzyme is a monomer . In the presence of NADPH Glu-tRNA reductase catalyzed the reduction to GSA of glutamate acylated to the homologous tRNA . Thus, the reductase alone is sufficient for conversion of Glu-tRNA to GSA . In the absence of NADPH, a stable complex of Glu-tRNA reductase with Glu-tRNA can be isolated. Nor Tannlaegeforen Tid, 1990 Mar, 100(4), 140 - 3 {Xylitol, mechanisms of action and uses}; Waler SM et al.; Xylitol is recommended as a sugar substitute and is claimed not only to be non-cariogenic, but also to exhibit an anti-caries effect . An interesting aspect of xylitol is that it has a certain bacteriostatic effect . Xylitol is taken up by many strains of Strep mutans and Strep sanguis even if these organisms are unable to metabolize xylitol . It enters the bacteria by the phospho-transferase system . Xylitol-phosphate inhibits the glycolysis probably at the phosphofructokinase level, and is also de-phosphorylated and expelled through a "futile cycle" . The combination xylitol and sorbitol is particularly interesting, since xylitol inhibits the metabolization of sorbitol by Strep mutans and sorbitol also appears to potentiate the bacteriostatic effect of xylitol . Furthermore the combination is favourable from an economical point of view since sorbitol is less expensive than xylitol . Xylitol induces diarrhea if consumed in considerable amounts and is thus only suitable in products like chewing gum and tablets. Boll Soc Ital Biol Sper, 1990 Mar, 66(3), 303 - 8 {Scanning electronic microscopy study of nasopharyngeal carcinoma}; Dallai S et al.; The nasopharyngeal carcinoma is a greatly interesting model of study . Its morphology has been studied in 8 patients affected by squamous cells carcinoma (4 cases) and by undifferentiated carcinoma (4 cases) . By means of a fibre-optic pharyngoscope equipped with mechanical biopsy forceps, biopsies have been taken and subsequently studied by scanning electron microscopy (SEM) . Squamous cells carcinoma is characterized by large, flattened elements, generally equipped with a network of microridges . Irregular microvilli or smooth apices have been observed too . Desquamating elements often accumulated in waves or vortices, as far as little cornified structures have been described . Undifferentiated carcinoma shows a great morpho-structural disorder . In a rugged surface, it is possible to observe element highly varying for size and shape . The apical plasmalemma is characterized by pleomorphic microvilli, while it must be emphasized the presence of little pseudopod-like extrusions of some cellular apices . Ciliated elements have been observed too, whose apical specializations are distorted or irregularly disposed . Necrotic zones are particularly frequent, in which bacteria or mycetes proliferate . The role of SEM in the study of this tumour is emphasized, because of the close relation between epithelial patterns and natural history of nasopharyngeal carcinoma. Adv Contracept, 1990 Mar, 6(1), 41 - 5 The correlation between Chlamydia antigen, antibody, vaginal colonization and contraceptive method in young unmarried women; Blum M et al.; In a group of 55 unmarried women, mean age 25 years, attending a family planning clinic and having minor gynecological complaints, the correlation between Chlamydia trachomatis (CT) antigen, CT antibodies, vaginal colonization by Candida or bacteria and the method of contraception was investigated . The correlation between CT antigen and CT antibodies (IgG) was significant in oral contraceptive users (p = 0.003), as was the correlation with vaginal colonization by Candida and potential pathologic bacteria . In the group using the natural family planning method, a statistically significant correlation was found between CT antigen, IgG (p = 0.002), IgA (p = 0.02) antibodies, and vaginal candidiasis (p = 0.002), but not with bacterial colonization (p = 0.90) . The discrepancy between CT antigen and antibodies is discussed . Differences in the prevalence of Chlamydia trachomatis infection were found among groups using different birth control methods, indicating an association between Chlamydia infection and the contraceptive method used. East Afr Med J, 1990 Mar, 67(3), 172 - 81 Perioperative infections in Nigerians: a seven-year prospective study; Odelowo EO et al.; The introduction of simple, measures in addition to usual aseptic and antiseptic measures at the University of Ilorin Teaching Hospital theatre in Ilorin, Nigeria, led to a significant decrease in perioperative infection rate in a pilot study . These measures were applied to 440 operative procedures in a unit over a 7-year period in an old as well as new theatre . Sterility tests on sterilizing packs, nasal and throat swabs and bacteria-carrying particle samplings were done to document and limit the sources of wound contamination . Mortality and infection rates were significantly higher among patients undergoing thoracic than extrathoracic surgical operation (P less than 0.05; p less than 0.005) including post-tube thoracostomy empyema . Clean and clean-contaminated cases survived operations significantly more frequently and were significantly less infected than the contaminated and dirty cases (less than 0.001; p less than 0.005) . Although there was significantly higher mortality (p less than 0.05) in patients older than 31 years, there was no significantly higher infection rate . Neither the mortality rate nor infection rate was significantly affected by seasonal and patients' sex . Overall infection rate was 7.5% (32 out of 428 fully evaluated patients) while wound and non-wound infection rate in this study is an improvement over those previously reported in this country. J Appl Bacteriol, 1990 Mar, 68(3), 225 - 30 Electronic particle counting for evaluating the quality of air in operating theatres: a potential basis for standards? Seal DV, Clark RP. Airborne particle counting in eight size ranges (0.5- greater than 20 microns), by computerized electronic equipment, was compared with the numbers of bacteria-carrying particles (BCP) assessed by slit sampling in ultra-clean and turbulently ventilated operating theatres . In the ultra-clean theatre the number of particles of 5-7 microns size range correlated with BCP while peaks in the numbers of particles less than 3 microns and greater than 15 microns corresponded with activity . Comparative relationships also occurred in the turbulently ventilated theatre but the use of this equipment in that environment cannot yet replace counts of airborne bacteria . We consider that electronic particle counting in the 0-20 microns size range may be used to judge the performance of a clean air operating theatre distribution system, including efficiency and integrity of the filter/seal systems and the presence or absence of entrainment of bacteria and other particles . The sampling techniques and analysis of particle concentration results described here may be a suitable basis for standards. Nuklearmedizin, 1990 Mar, 29(2), 54 - 8 99mTc-labelled polyclonal human immunoglobulin for localization of inflammatory sites--early in vitro-results; Saptogino A et al.; 99mTc-labelled polyclonal human immunoglobulin (HIG) has been shown to be able to localize inflammatory sites . Hypothetically these immunoglobulins bind directly to bacteria, HIG binds directly to Fc-receptors on the surface membrane of leucocytes or HIG simply passes vessels of increased permeability . To collect further information experiments were carried out in vitro with the blood of human volunteers . 0.5 mg 99mTc-HIG, 2 mg 99mTc-human albumin and 0.5 mg 99mTc-labelled murine monoclonal antigranulocyte antibodies were added to in vitro isolated human "mixed" leucocyte pellets and to 30 ml whole blood of 6 healthy volunteers . The whole blood and the directly labelled leucocyte pellet were layered and separated on a discontinuous Percoll/plasma gradient . The activity distribution was measured within the gradient . The 99mTc-HIG labelled gradients showed a significant uptake of the activity within the monocyte band whereas the 99mTc-albumin gradients showed no specific albumin uptake in any cellular band . The 99mTc-antibody labelled gradients showed a significantly increased uptake on granulocytes . It is concluded that in man a specific monocyte-associated uptake and binding mechanism of 99mTc-HIG plays an important role in the localisation of inflamed sites. J Antimicrob Chemother, 1990 Mar, 25(3), 407 - 12 Pharmacokinetics of imipenem/cilastatin in neutropenic patients with haematological malignancies; Janmohamed RM et al.; The pharmacokinetics of imipenem/cilastatin were studied in febrile neutropenic patients with haematological malignancies . The peak plasma concentrations (36.4 +/- 4.96 mg/l), plasma half-life (60 min), volume of distribution (0.28 +/- 0.02 l/kg) and plasma clearance (3.23 +/- 0.38 ml/min/kg) were comparable with those in normal healthy volunteers suggesting that the drug handling is not appreciably altered in this group of patients . The administration of 12.5 mg/kg (max 1 g), 6-hourly achieved levels that were up to 3.5 times MICs of most relevant bacteria . The drug therefore has a potential use as empirical monotherapy in febrile neutropenic patients. J Burn Care Rehabil, 1990 Mar-Apr, 11(2), 146 - 50 Is there a need for state health department sanitary codes for public hydrotherapy and swimming pools? Zura RD, Groschel DH, Becker DG, Hwang JC, Edlich RF. The Board of Health of the Commonwealth of Virginia has an outdated sanitary code for its public hydrotherapy and swimming pools . The code is restricted to pools in hotels and other lodging places . The absence of modern regulations for public hydrotherapy and swimming pools has permitted serious deficiencies in pool maintenance, which are highlighted in this report . The most notable of these deficiencies was the presence of high levels of bacterial contamination that could predispose to infect in the water of one public hot tub . The results of this study indicate that the Virginia Board of Health sanitary code for pool water must be revised immediately and should include all public hydrotherapy and swimming pools . Other states and communities may want to assess their codes for swimming pools and hydrotherapy tubs to avoid deficiencies that could be detrimental to public health. Gene, 1990 Mar 1, 87(1), 105 - 12 Sulfur-rich proteins of Chlamydia trachomatis: developmentally regulated transcription of polycistronic mRNA from tandem promoters; Lambden PR et al.; RNA was extracted at various times from cells infected with Chlamydia trachomatis serovar L1 . Northern-blot analysis showed that transcription of the CrP gene encoding the 60-kDa cysteine-rich outer membrane protein (CrP) produces a temporally controlled polycistronic mRNA . Primer extension analysis indicated the presence of tandem promoters separated by 66 nt with transcriptional start points (tsp) located 577 and 643 nt upstream from the start codon of the mature 60-kDa CrP . Nucleotide (nt) sequencing of this region revealed a small open reading frame (SORF) with coding potential for an 88-amino acid protein containing 13 cysteine residues . This SORF is transcribed as both a polycistronic 2300-nt mRNA together with the CrP gene, and as a separate 480-nt mRNA . Analysis of the upstream sequences, around the tsp for these mRNAs, revealed the presence of three inverted repeat structures that might act as binding domain(s) for a regulatory protein. No To Hattatsu, 1990 Mar, 22(2), 179 - 83 {A case of neonatal herpes simplex virus encephalitis with calcifications of thalamus and basal ganglia}; Takahashi H et al.; A male infant developed left-sided myoclonus associated with low grade fever at 21 days of age . The course of pregnancy and delivery had been uneventful . Birth weight was 2,240g at 38 weeks of gestation . Physical examination at 26 days of age revealed hyperirritability and incomplete Moro reflex . EEG showed periodic discharges originating in the right central region . Initial brain CT at 26 days of age showed no apparent abnormality . In lumbar CSF protein was 173 mg/dl, and nucleated cells 328/mm3 (polynuclear cells 6 and mononuclear cells 322) . Cultures of CSF for bacteria and viruses were negative . Although, the serum titers of CF antibody to herpes simplex virus (HSV) were 1:4 to 1:8 during hospitalization and those in CSF were not increased significantly, ELISA titers for IgG and IgM in serum and CSF rose significantly . The diagnosis of HSV encephalitis was made at 41 days of age . His myoclonus was well controlled with phenobarbital and clonazepam . He was treated with acyclovir and CSF findings returned to normal . On 2nd brain CT (39 days of age), calcification at thalamus, basal ganglia and right frontal lobe, which is extremely rare CT findings for HSV encephalitis, was noticed . His DQ at discharge was about 90 . It is emphasized that the early diagnosis of HSV encephalitis can be made by ELISA and serial brain CT examinations, and that HSV encephalitis should be considered when calcification of thalamus and basal ganglia are detected on brain CT. J Am Mosq Control Assoc, 1990 Mar, 6(1), 35 - 42 Natural food and feeding behavior of Coquillettidia perturbans larvae; Merritt RW et al.; The natural particulate food of larval Coquillettidia perturbans was studied through gut analysis using staining with 4'6-diamidino-2-phenylindole and epifluorescence microscopy . Bacteria (cocci, rods, spirochetes, purple bacteria and cyanobacteria), detritus, euglenoid protozoans and algae (desmids and diatoms) comprised the majority of particulate food, in order of abundance; other protozoans were rare, and hyphal forms (actinomycetes and fungi) were not observed . Abundance of food items in guts did not vary greatly among sampling months (June-December, 1987) or sampling sites . Fourth instars had a greater proportion of euglenoids and algae in their food than did younger instars . Observation of larval feeding behavior showed that fourth instars oriented upside down and fed mainly by suspension feeding, with occasional brushing from sediments and root surfaces . Microtrichia on filaments of the lateral palatal brushes of fourth instars indicated that the larvae were adapted to collect fine particles. J Clin Periodontol, 1990 Mar, 17(3), 138 - 44 The influence of surface free energy and surface roughness on early plaque formation . An in vivo study in man; Quirynen M et al.; Previous in vivo studies suggested that a high substratum surface free energy (s.f.e.) and an increased surface roughness facilitate the supragingival plaque accumulation . It is the aim of this clinical trial to explore the "relative" effect of a combination of these surface characteristics on plaque growth . 2 strips, one made of fluorethylenepropylene (FEP) and the other made of cellulose acetate (CA) (polymers with surface free energies of 20 and 58 erg/cm2, respectively) were stuck to the labial surface of the central incisors of 16 volunteers . Half the surface of each strip was smooth (Ra +/- 0.1 microns) and the other half was rough (Ra +/- 2.2 microns) . The undisturbed plaque formation on these strips was followed over a period of 6 days . The plaque extension at day 3 and 6 was scored planimetrically from color slides . Finally, of 6 subjects samples were taken from the strips as well as from a neighbouring smooth tooth surface (s.f.e . 88 erg/cm2; Ra +/- 0.14 microns) . These samples were analysed with a light microscope to score the proportion of coccoid cells, and small, medium, and large rods or fusiform bacteria . At day 3, a significant difference in plaque accumulation was only obtained when a rough surface was compared with a smooth surface . However, at day 6, significantly less plaque was recorded on FEP smooth (19.4%) when compared with CA smooth (39.5%) . Between FEP rough (96.8%) and CA rough (98.2%), no significant difference appeared.(ABSTRACT TRUNCATED AT 250 WORDS) Clin Exp Immunol, 1990 Mar, 79(3), 463 - 9 Human milk contains proteins that stimulate and suppress T lymphocyte proliferation; Mincheva-Nilsson L et al.; The modulatory effect of human milk proteins from colostrum and late milk on the proliferative response of human T lymphocytes activated by mitogens (OKT3 and leucoagglutinin from Phaseolus vulgaris) and alloantigens was studied . High concentrations (10-100 micrograms/ml) of crude colostral milk proteins had an inhibitory effect on T cell growth while low concentrations (0.1-1 microgram/ml) enhanced T cells growth . In contrast, proteins from late milk did not inhibit T lymphocyte proliferation while the enhancing effect was retained . Colostrum was fractionated by ammonium sulphate precipitation and gel filtration on sepharose 6B . The inhibitory activity was recovered in a protein fraction containing lactoferrin as its major component . Lactoferrin was, however, not responsible for the observed inhibition . On the contrary, lactoferrin in most cases augmented the proliferative response induced by polyclonal activators . The inhibitory activity was found to bind concanavalin A-sepharose suggesting an association with glycoprotein . Inhibitory fractions contained glycoproteins of the following molecular sizes 26, 74/76 (doublet), 84, 145 and 160 kD under reducing conditions . The inhibitory effect appeared to be lymphocyte specific since the active fraction did not inhibit the growth of tissue culture cells (HeLa cells and human fibroblasts) or bacteria . Furthermore, the fraction was not toxic for lymphocytes . The inhibitory colostrum factor may prevent the newborn from overreacting immunologically against the environmental antigens encountered at birth. Cancer Lett, 1990 Mar, 49(3), 207 - 10 Modification of DNA by bile acids: a possible factor in the etiology of colon cancer; Cheah PY et al.; Bile acids have been implicated as promoters and cocarcinogens in the etiology of colon cancer and as comutagens and mutagens in bacteria . These observations suggest the hypothesis that bile acids may interact directly with DNA . We treated the single stranded circular DNA of phage M13 with bile acids and found that the transfection efficiency of this DNA declined up to a 1000-fold . This result suggests that bile acids can damage DNA and thus may play an important role in the etiology of colon cancer. Appl Environ Microbiol, 1990 Mar, 56(3), 739 - 46 DNA hybridization to compare species compositions of natural bacterioplankton assemblages; Lee S et al.; Little is known about the species composition and variability of natural bacterial communities, mostly because conventional identification requires pure cultures, but less than 1% of active natural bacteria are cultivable . This problem was circumvented by comparing species compositions via hybridization of total DNA of natural bacterioplankton communities for the estimation of the fraction of DNA in common between two samples (similarity) . DNA probes that were labeled with 35S by nick translation were hybridized to filter-bound DNA in a reciprocal fashion; similarities (in percent) were calculated by normalizing the values to self-hybridizations . In tests with DNA mixtures of pure cultures, the experimentally observed similarities agreed with expectations . However, reciprocal similarities (probe and target reversed) were often asymmetric, unlike those of DNA from single strains . This was due to the relative complexity and G + C content of DNA, which provided a means to interpret the asymmetry that was occasionally observed in natural samples . Natural bacteria were collected by filtration from Long Island Sound (LIS), N.Y., the Caribbean and Sargasso seas, and a coral reef lagoon near Bermuda . The samples showed similarities of less than 10 to 95% . The LIS and Sargasso and Caribbean sea samples were 20 to 50% similar to each other . The coral reef sample was less than 10% similar to the others, indicating its unique composition . Seasonality was also observed; an LIS sample obtained in the autumn was 40% similar to two LIS samples obtained in the summer; these latter two samples were 95% similar . We concluded that total DNA hybridization is a rapid, simple, and unbiased method for investigating the variation of bacterioplankton species composition over time and space, avoiding the need of culturing. J Bacteriol, 1990 Mar, 172(3), 1448 - 56 Attachment of the adhesive holdfast organelle to the cellular stalk of Caulobacter crescentus; Ong CJ et al.; Caulobacters attach to surfaces in the environment via their holdfasts, attachment organelles located at the base of the flagellum in swarmer cells and later at the end of the cellular stalk in the stalked cells which develop from the swarmer cells . There seems to be little specificity with respect to the types of surfaces to which holdfasts adhere . A notable exception is that the holdfast of one cell does not adhere to the cell surface of another caulobacter, except by joining holdfasts, typically forming "rosettes" of stalked cells . Thus, the localized adhesion of the holdfasts to the cells is in some way a specialized attachment . We investigated this holdfast-cell attachment by developing an adhesion screening assay and analyzing several mutants of Caulobacter crescentus CB2A selected to be defective in adhesion . One class of mutants made a normal holdfast by all available criteria, yet the attachment to the cell was very weak, such that the holdfast was readily shed . Another class of mutants made no holdfast at all, but when mixed with a wild-type strain, a mutant of this class participated in rosette formation . The mutant could also attach to the discarded holdfast produced by a shedding mutant . In addition, when rosettes composed of holdfast-defective and wild-type cells were examined, an increase in the number of holdfast-defective cells was correlated with a decrease in the ability of the holdfast material at the center of the rosette to bind colloidal gold particles . Gold particles are one type of surface to which holdfasts adhere well, suggesting that the stalk end and the colloidal gold particles occupy the same sites on the holdfast substance . Taken together, the data support the interpretation that there is a specialized attachment site for the holdfast at the base of the flagellum which later becomes the end of the stalk, but not a specialized region of the holdfast for attachment to this site . Also, attachment to the cell is accomplished by bond formations that occur not only at the time of holdfast production . Thus, we propose that the attachment of the holdfast to the cell is a true adhesion process and that the stalk tip and base of the flagellum must have compositions distinctly different from that of the remainder of the caulobacter cell surface. J Bacteriol, 1990 Mar, 172(3), 1430 - 5 Generation of a membrane potential by sodium-dependent succinate efflux in Selenomonas ruminantium; Michel TA et al.; When Selenomonas ruminantium HD4 was grown in a chemostat, maximal succinate production and the highest molar growth yield values were both observed at a dilution rate of roughly 0.2 h-1 . To determine the possible relationship between succinate efflux and high molar growth yields, the generation of a membrane potential by succinate efflux was studied in whole cells and vesicles (inside-out and right-side-out) prepared from S . ruminantium . Washed whole cells took up succinate in the absence of an exogenous energy supply; uptake was completely abolished by brief treatment with dinitrophenol or with nigericin and valinomycin . High levels of sodium ions (with respect to the intracellular sodium concentration in the assay buffer had a stimulatory effect on succinate uptake . When succinate was added to inside-out vesicles, a membrane potential (inside positive) was generated, as indicated by fluorescence quenching of the anionic lipophilic dye Oxonol V . Fluorescence quenching was sensitive to uncoupling by gramicidin D but only partially sensitive to the uncoupler carbonyl cyanide-p-trifluoromethoxyphenylhydrazone . In right-side-out vesicles, succinate uptake could be driven by an artificially imposed sodium gradient but not by a potassium diffusion potential; imposition of both a sodium gradient and potassium diffusion potential resulted in improved succinate uptake . The generation of a membrane potential (inside negative) upon succinate efflux was demonstrated directly in right-side-out vesicles when succinate-loaded vesicles were diluted into succinate-free buffer, and the lipophilic cationic probe tetraphenylphosphonium accumulated in the vesicles . Results indicate that an electrogenic succinate-sodium symporter is present in S . ruminantium . Transport of succinate out of the cell via the symporter might be responsible for the high molar growth yields obtained by this organism when it is grown at dilution rates where maximal succinate production occurs. Geriatrics, 1990 Mar, 45(3), 76 - 7, 80 When 'something is wrong': a case for investigating vague health complaints; Fader AM et al.; When a chronically ill elderly woman complained that she felt something was wrong, close attention and follow-up by physician and nursing home staff paid off. Res Microbiol, 1990 Mar-Apr, 141(3), 286 - 9 Osmochemistry of solute translocation; Mitchell P; The main aim of this brief contribution is to suggest that our understanding of the general principles of osmochemistry may provide useful insights into the type of mechanism by which solute-translocating catalysts work . In particular, I would like to encourage a more widespread and explicit recognition of the special merits of the mobile barrier type of mechanism (Mitchell, 1957, 1987), not as a panacea, but to explain the translocation of the characteristically hydrophilic and somewhat bulky solutes that are the main substrates of solute porters and of some osmoenzymes in bacterial membranes. Nihon Kyobu Shikkan Gakkai Zasshi, 1990 Mar, 28(3), 511 - 8 {Second serogroup of Legionella pneumophila isolated from a patient with fulminant pneumonia}; Tomioka H et al.; A 67-year-old female was admitted to our hospital, because of high fever and dry cough . She had undergone semiradical hysterectomy and radiation therapy for carcinoma of the uterine cervix one year previously . Her chest roentgenograms on admission showed lobar consolidation of the left upper lobe . Antibiotics were administered but her general condition and pulmonary consolidation did not improve . As Legionellosis was highly suspected, we performed bronchoscopic examinations . Bronchial mucosa was almost normal with no secretion, and Legionella was isolated from the specimen obtained bronchoscopically . Six days later, L . pneumophila serogroup 2 was isolated and identified from an intratracheal aspiration, and serological diagnosis was made by indirect immunofluorescence antibody . We could also detect the bacteria in the BALF by immunofluorescence microscopy and in the tissue of the TBLB specimen with the ABC method. J Cell Sci, 1990 Mar, 95 ( Pt 3), 487 - 98 Polymorphonuclear leukocyte phagocytosis of Capnocytophaga ochracea in three-dimensional plasma clots; Hurst TJ et al.; We have studied the ability of human polymorphonuclear leukocytes (PMN) to phagocytose Capnocytophaga ochracea in three-dimensional fibrin meshworks . Phagocytosis was assessed in three systems: (1) the PMN and bacteria were mixed together with plasma and clotted; 60 +/- 13% phagocytosis occurred after 60 min; (2) PMN were overlaid on clots containing bacteria; the PMN migrated into the clot and after 60 min 52 +/- 7% phagocytosis was seen; (3) PMN had to migrate from within one clot into a second containing bacteria; phagocytosis after 60 min was 54 +/- 3% . In the clots, PMN released lysozyme but this was not significantly enhanced by phagocytosis . These findings indicate that PMN are capable of phagocytosing in each of the three-dimensional systems tested and that they are capable of both migration into and subsequent phagocytosis in a model that more closely mimics the in vivo structure in which PMN would normally perform. Ann Pediatr (Paris), 1990 Mar, 37(3), 167 - 8 {Knee arthritis caused by Eikenella corrodens . Apropos of a case}; Beauvais P et al.; We report a case of septic arthritis of the knee following an articular wound caused by a bite in a previously healthy 8-year-old girl . The causative agent was Eikenella corrodens, an opportunistic bacteria that is normally part of the commensal flora of the oral cavity . We point out that Eikenella corrodens may cause severe complications in healthy subjects and emphasize the need for considering this organism not only in infections of the hands, face and neck, but also after bites . Penicillins are consistently effective and constitute the treatment of choice. Can J Microbiol, 1990 Mar, 36(3), 231 - 5 Comparison of different membranes for use in the colony-immunoblot technique; Rodrigue L et al.; We compared five different supports (Whatman paper filters Nos . 1, 5, and 40, nitrocellulose, and Nylon 66) for their suitability in the colony-immunoblot (CIB) technique . Results indicate that Whatman No . 5 filter paper recovered 94-98% of the bacterial colonies tested, were more resistant to tearing than the other Whatman papers tested, and showed reduced cross-reactions as compared with nitrocellulose membranes . Whatman No . 5 filters are 20 times less expensive than the nitrocellulose membranes usually used in the CIB technique . We thus adopted the former for our ecological studies of the murine oral cavity. Zentralbl Bakteriol, 1990 Mar, 272(3), 265 - 75 Relationship between free amoeba and Legionella: studies in vitro and in vivo; Vandenesch F et al.; In 1980, Robowtham demonstrated that Legionella multiplies in free amoeba cytoplasm and hypothesized that the amoeba could act as a reservoir of virulent bacteria . In this paper we report various aspects of the relationship between amoeba and Legionella . A liquid medium co-culture method was applied to Acanthamoeba sp . and Legionella pneumophila serogroup 1 . Within 4 days, Legionella growth increased by 2 log s CFU/ml . Using a direct immunofluorescence assay and electron microscopy, Legionella was shown to grow abundantly inside phagosomes, and bacteria and/or antigen were present on the cytoplasmic membrane of the amoeba . These aspects are very similar to those observed with Legionella-infected alveolar macrophages . The morphology and structure of Legionella cells were modified after 20 days of co-culture: - viable bacteria showed large fatty cytoplasmic inclusions, - gas liquid chromatography analysis demonstrated a decrease in the i16:0 fatty acid ratio . Cystic forms of amoeba were abundant but none contained viable Legionella . In an in-vivo study using a guinea-pig aerosol infection model, we compared the virulence of Legionella in co-culture with Legionella grown on charcoal dialysed yeast extract (CDYE) agar medium . The Legionella obtained by co-culture had an LD 50 (50% lethal dose) similar to that obtained for those grown on CDYE, showing that bacterial virulence is preserved in the cellular model. Berl Munch Tierarztl Wochenschr, 1990 Mar 1, 103(3), 88 - 90 {The persistence of bovine enterovirus and pseudorabies virus in liquid cattle manure at different storage temperatures}; Biermann U et al.; The tenacity of viruses in liquid manure of cattle was examined in a total of five samples inoculated with ECBO-virus (strain LCR-4) representing viruses without envelope and Aujeszky virus (field isolate) representing enveloped viruses . The titers were examined at regular intervals over a period of 26 weeks . On the day of inoculation each sample had a titer of 10(5) ID50/ml . After 16 weeks complete inactivation was observed in the Aujeszky virus sample stored at 20 degrees C . The Aujeszky virus sample wich was kept at 4 degrees C at 26 weeks had a titer of 10(1,75) ID50/ml . In the samples inoculated with ECBO virus after 26 weeks of inoculation a titer of 10(3) ID50/ml was found in the manure stored at 20 degrees C . No influence on the virus titers in the liquid manure samples was observed either from pH or the number of bacteria (3,4 x 10(7)-1.16 x 10(8)/ml during the examination period. J Bacteriol, 1990 Mar, 172(3), 1262 - 6 Circularity of the Caulobacter crescentus chromosome determined by pulsed-field gel electrophoresis; Ely B et al.; Previous genetic analyses of the Caulobacter crescentus chromosome have resulted in the construction of a linear genetic map . To establish the circularity of the C . crescentus chromosome, restriction fragments generated by digestion with AseI and SpeI were analyzed by pulsed-field gel electrophoresis and Southern hybridization . The size of each fragment was calculated and used to demonstrate that C . crescentus has a genome size of approximately 4,000 kilobases . In addition, both enzymes gave rise to large DNA fragments which contained genes from both ends of the genetic map . Thus, there is physical linkage between the genes at the ends of the genetic map and the chromosome is circular . Since this region of the chromosome appears to contain the replication terminus, we propose that recombination occurs at a high frequency in the vicinity of the terminus . This high frequency of recombination would prevent genetic linkage from being observed between genes on opposite sides of the terminus . Additional experiments using insertions which introduced new AseI and DraI restriction sites into the genome allowed us to calculate the physical distance between genes located in the vicinity of the replication terminus. Nichidai Koko Kagaku, 1990 Mar, 16(1), 16 - 26 {Stratification of the experimental incipient caries (polarized light microscopy and microradiography)}; Kawasaki M et al.; The enamel caries establishes when at least a part of enamel dissolves by the action of cariogenic bacteria which produce insoluble glucan and lactic acid . The experimental caries should be prepared at the condition which simulates the natural environment . The author made 1 mm thick sections from 29 teeth extracted for convenience of orthodontic reason . The surface was covered except the "window" which will be exposed to the acid milieu . The samples were immersed for 4 to 120 days in the gelatin added lactate buffer of pH 4.5, pH 5.0, and pH 5.5 . Ground sections were made after the experiment . The structure of the experimental incipient caries was described after Darling (1963); surface layer, body of the lesion, dark zone, and translucent zone . When the section was immersed for shorter period in the buffer of pH 4.5, the lesion was made up only two layers, namely, the surface layer and the body of the lesion . This type of lesion will be called Type I, hereafter . When the action of acid was more intense, only one layer, the body of the lesion remained after the dissolution of the surface layer . This type of lesion will be called Type II . The lesion which provided with all 4 layers will be called as Type III, and the lesion which was made up of 3 layers which remained after dissolved surface layer, will be called as Type IV . Both Type III and Type IV were established after immersing for longer period in pH 5.5, although they could be observed in groups which immersed in lower pH . Of the samples examined, a few example showed a five-layered structure; the surface layer, the first body of lesion, the first dark zone, the second body of lesion, the second dark zone, and the translucent zone . This phenomenon suggests that the speed of caries development was not uniform throughout the experiment . The relationship between the depth of the lesion, and the pH and the duration of acid action was studied . In the pH 5.0 group, the development of caries lesion was temporarily arrested between 30 and 60 days . This phenomenon would be an explanation to the formation of five-layered lesion. Zhonghua Kou Qiang Yi Xue Za Zhi, 1990 Mar, 25(2), 82 - 5, 125 {Clinical pathological analysis of recurrent oral ulcer and Behçet's syndrome}; Gao C; Histopathological features of 53 cases of recurrent oral ulcer (ROU) and 28 cases of Behcet's syndrome (BS) were studied . The findings of light microscopic examinations are as follows: (1) The inflammatory cell reactions in early lesions are characterized by predominant lymphocytes and monocytes which differ from those in later lesions . (2) The vascular lesions are more significant in BS than in ROU . (3) The number of mast cells in the oral lesions of BS is more than that in ROU and controls . (4) Bacteria are observed in 11 specimens stained with HE and multinucleated epithelial giant cells are found in one specimen . It is considered that the lymphomonocytes in epithelial layer may play a role in the destruction of the epithelial cells through autoimmunization. Zentralbl Veterinarmed A, 1990 Mar, 37(2), 148 - 53 Immunological induction of endometritis-model investigations in cows; Klucinski W et al.; The course of the inflammatory reaction in the uterus after local or systemic administration of specific or non-specific antigens was studied . The study was carried out on 40 cows, 10-11 weeks after labour and between 6 and 8 days after ovulation . A specific cell-mediated type of reaction was induced by intrauterine challenge with PPD in cows previously vaccinated with M . bovis subcutaneously or by intrauterine administration . A specific inflammatory process of the type of Arthus reaction was induced by intrauterine challenge with C . fetus ssp . veneralis in animals immunized with these bacteria subcutaneously or by intrauterine instillation of these bacteria . A non-specific inflammatory process in the uterus was initiated by one instillation of lipopolysaccharide (LPS) into the left uterine horn . The cells were washed out from the uterus before and 6, 24, 48 and 72 hours after initiation of the specific or non-specific inflammatory process . The per cent proportions of various cell types were determined . It was demonstrated that intrauterine instillation of specific or non-specific antigens in cows caused a significant rise in PMNs per cent in relation to the control group. Urol Nefrol (Mosk), 1990 Mar-Apr, (2), 17 - 20 {The differential diagnosis of allergic prostatitis}; Lopatkin NA et al.; The leading role of allergic components was established in some rare patterns of prostatitis . To develop the methods applicable for differential diagnosis of allergic prostatitis, the authors followed up 60 patients aged from 15 to 53 years who suffered from chronic prostatitis and 10 normal controls . The patients with bacterial disease, abacterial prostatitis with concurrent allergic components or subjects with allergic prostatitis were enrolled in the study . Immunoassay was an essential part in the diagnosis of allergic prostatitis . The study revealed a dramatic inhibition of ejaculate gamma-glutamine transferase in those who suffered from an abacterial pattern with allergic component and almost three-fold in those with allergic prostatitis . Certain though insignificant decrease in the ejaculate gamma-glutamine transferase was documented in patients with bacterial prostatitis . Investigations of ejaculate IgE revealed quite an opposite picture: 10-fold increase in IgE levels in the ejaculate of patients with allergic prostatitis as compared with other groups, controls in their number . Besides, patients with allergic prostatitis had three-fold higher IgE in the ejaculate than in the blood serum . The aforementioned phenomenon failed to be observed in patients with other patterns of prostatitis . Decreased ejaculate gamma-glutamine transferase and increased IgE can be regarded as criteria for differential diagnosis of allergic prostatitis. Rev Sci Tech, 1990 Mar, 9(1), 245 - 51 Use of biotechnical methods in veterinary medicine; Haas L et al.; Biotechnological methods offer promising approaches for improved diagnostic and prophylactic purposes . The following biotechnological techniques are used in the Institute of Virology at the Hanover Veterinary School:--Production of monoclonal antibodies directed against viral and bacteria-specific antigens such as bovine virus diarrhoea virus, classical swine fever (hog cholera) virus, feline leukaemia virus, animal parvoviruses, Alphavirus, Brucella and Francisella--Establishment of improved and sensitive diagnostic enzyme immunoassays (ELISA) using monoclonal antibodies--Molecular cloning and sequencing of classical swine fever virus RNA and parvovirus DNA--Development of diagnostic hybridisation techniques (dot, slot, Southern and Northern blot, in situ, oligonucleotides)--Detection of viral genomes in tissues of infected animals--Development of synthetic oligopeptides as diagnostic antigens and as potential immunogens for vaccines . Currently available techniques used in basic research (e.g . pathogenesis studies) will be tested for their application in routine diagnosis of viral diseases, e.g . by molecular hybridisation . Some techniques need to be simplified (e.g . RNA extraction procedures) and, particularly, alternative labelling schedules must be developed (e.g . biotin or sulfone labelling instead of radionuclides). Mol Gen Genet, 1990 Mar, 221(1), 65 - 71 Identification and characterization of a defective SSV1 genome integrated into a tRNA gene in the archaebacterium Sulfolobus sp . B12; Reiter WD et al.; Within the chromosome of the archaebacterium Sulfolobus sp . B12, a 7.4 kb region was identified which displayed extensive sequence similarities to the 15.5 kb genetic element SSV1 carried by the same strain both as a circular form and as a site-specifically integrated copy . DNA sequence analysis indicated that this 7.4 kb region (designated SSV1intB) represented an SSV1-like element distinguishable from the full-length integrated copy (designated SSV1intA) by extensive deletions and point mutations . The physical organization of DNA sequences of SSV1intB indicated that this element was integrated at the same attP site as previously identified for SSV1intA . A comparison of the DNA sequences at the left attachment sites of SSV1intA and SSV1intB revealed that they both represented very similar putative arginine tRNA genes followed by a 10 bp inverted repeat sequence . S1 nuclease mapping experiments indicated that these tRNA genes are transcribed. J Clin Microbiol, 1990 Mar, 28(3), 463 - 8 Demonstration of heterogeneity among the antigenic proteins of Mobiluncus species; Schwebke JR et al.; The protein and antigenic profiles of the American Type Culture Collection type strains of Mobiluncus species and those of 114 clinical isolates were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and immunoblotting with homologous polyvalent antisera . The majority of isolates (82%) possessed characteristic protein profiles and could be identified to the species level by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis . The major protein bands were also antigenic, and some antigenic cross-reactivity was noted between the two Mobiluncus species . All of the isolates were examined for reactivity with a panel of 12 monoclonal antibodies previously prepared against the type strains . While 56 of 60 clinical isolates of Mobiluncus curtisii (93%) reacted with one or more of the monoclonal antibodies, only 23 of 54 clinical isolates which were identified as Mobiluncus mulieris by biochemical methods (48%) reacted with one or more of the monoclonal antibodies . One of the 4 M . curtisii isolates (25%) and 11 of the 31 M . mulieris isolates (35%) which did not react with the monoclonal antibodies also had atypical protein profiles . These results demonstrate a high degree of heterogeneity in the protein and antigenic profiles of Mobiluncus isolates and suggest that further taxonomic division may be appropriate. J Exp Med, 1990 Mar 1, 171(3), 831 - 41 Recognition of a mycobacteria-specific epitope in the 65-kD heat-shock protein by synovial fluid-derived T cell clones; Gaston JS et al.; Adjuvant arthritis in rats is induced by a T cell clone specific for amino acids 180-188 of the mycobacterial 65-kD heat-shock protein, and synovial T cell responses to this same Ag have been noted in human arthritis . We have isolated 65-kD Ag-specific T cell clones from synovial fluid mononuclear cells of a patient with acute arthritis, which, unlike the corresponding PBMC, showed a marked proliferative response to the 65-kD Ag . Using synthetic peptides corresponding to the whole sequence of the 65-kD Ag, all the clones were shown to recognize an epitope present in the first NH2-terminal peptide (amino acids 1-15), with no response to the adjacent peptide (amino acids 6-22) or to any other peptide . The complete dominance of this epitope in the response to the 65-kD Ag was shown by documenting responses to the peptide in PBMC obtained after recovery from the arthritis . This epitope, like that recognized by the rat arthritogenic T cell clone, is in a portion of the 65-kD sequence that is not conserved between bacteria and eukaryotes, so that in this case, joint inflammation could not be attributed to bacteria-induced T cell clones cross-reacting with the self 65-kD Ag. Biochem Biophys Res Commun, 1990 Feb 28, 167(1), 89 - 95 Isolation, preliminary chemical characterization, and biological activity of Borrelia burgdorferi peptidoglycan; Beck G et al.; Peptidoglycan (PG), an essential cell wall polymer of most bacteria, has been isolated from many species of spirochetes . Our interest in the host response to Borrelia burgdorferi led us to isolate and characterize its PG . Extracted cells were solubilized with warm 1% SDS followed by digestion with proteases . Amino acid analysis of the isolated PG demonstrated the presence of alanine, glycine, glutamic acid, and ornithine as occurs in other spirochetes and bacteria . Intense erythematous reactions were observed after id injection of 10 micrograms of PG into normal human skin . PG was not mitogenic for human peripheral blood mononuclear cells . Murine splenocytes of certain strains responded to the PG, but only at concentrations of 25 micrograms/ml or more . PG stimulated macrophages to produce interleukin 1 . Sixteen micrograms of PG injected iv into rabbits produced biphasic fevers . These observations on the in vitro and in vivo activities associated with the cellular components of the B . burgdorferi spirochete give further insight to how a small number of invading organisms can cause a multisystemic disease such as Lyme disease. Ned Tijdschr Geneeskd, 1990 Feb 24, 134(8), 380 - 5 {Maxillary sinusitis in children}; van Buchem FL et al.; In children with nasal discharge the distinction between the group with a diagnosis of rhinitis and the group with a diagnosis of sinusitis was vague . Anamnestic data and findings at physical examination were not significantly different . There was hardly any relation between radiographic and echographic findings, the nature of the irrigation fluid, and isolation of pathogenic or non-pathogenic bacteria from cultures of nasal or sinus secretions . In our opinion there is no clearcut difference between rhinitis and simple sinusitis (i.e . mucositis of the maxillary sinus without empyema) . Nor is this therapeutically relevant because both conditions are mostly features of a respiratory tract infection . It is therapeutically important, however, to differentiate between sinusitis without and sinusitis with empyema . The latter requires special treatment . None of the patients studied had a sinusitis with empyema . When a child has running nose there is no reason to look for a sinusitis when there are no clinical symptoms of empyema. Arch Biochem Biophys, 1990 Feb 15, 277(1), 74 - 9 Evidence for superoxide dismutase and catalase in mollicutes and release of reactive oxygen species; Meier B et al.; The presence of superoxide dismutase was demonstrated in 21 strains of mollicutes, including achuloplasmas, mycoplasmas, and ureaplasmas . No superoxide dismutase activities or only traces were detectable in fresh prepared cell lysates, whereas activities were evident after dialysis of the cell lysates . A further increase in superoxide dismutase activities was observed after the cell lysates were heated to 65 degrees C for 30 s . This might be due to the destruction of enzymatic reactions interfering with the activity tests . Additionally, catalase activities were demonstrated in nearly 50% of the cell lysates, whereas no peroxidase activities were detectable . The production of O2- and H2O2 with glucose as substrate was demonstrated for 8 of 10 strains tested . No correlation to the pathogenicities of the strains was indicated . Anaerobic mycoplasmas showed the highest amount of radical production, whereas superoxide dismutase and catalase activities were in the range of activities estimated for aerobic mollicutes. Am J Ophthalmol, 1990 Feb 15, 109(2), 138 - 42 A systematic approach to the diagnosis and treatment of chronic conjunctivitis; Rapoza PA et al.; In 58 patients with chronic conjunctivitis of greater than two weeks' duration, examination included obtaining an ocular and general medical history and performing a complete ophthalmic examination of the external eye . Conjunctival smears were obtained for Gram and Giemsa staining, direct immunofluorescent monoclonal antibody staining for Chlamydia trachomatis and herpes simplex virus, and chlamydial culture . Cultures for bacteria and viruses were obtained in 33 patients . The cause of the chronic conjunctivitis based on clinical and laboratory criteria was established in 40 of 58 (69%) patients: chlamydia, 11 (19%); virus, eight (14%); irritant, six (10%); allergen, four (7%); contact lens, four (7%); bacteria, four (7%); acne rosacea, two (3%); and floppy eyelid syndrome, one (2%) . In 18 of 58 (31%) patients, no specific cause was detected . We recommend a systematic approach in the investigation of chronic conjunctivitis . Direct immunofluorescent monoclonal antibody staining is an effective and rapid technique for detecting chronic chlamydial conjunctivitis. J Biol Chem, 1990 Feb 15, 265(5), 2988 - 92 Histidine tRNA from chicken mitochondria has an uncoded 5'-terminal guanylate residue; L'Abbe D et al.; In an attempt to identify the transcription initiation sites in chicken mitochondrial DNA, RNAs capped in vitro using vaccinia guanylyl transferase and {alpha-32P} GTP were analyzed . The most abundant labeled transcript was identified by RNA sequencing as the mitochondrial tRNA(His) . Sequence analysis also revealed that this tRNA contains an extra guanylate residue at its 5' end, characteristic of the histidine tRNA family . The respective genomic region was also cloned and sequenced . In contrast to bacteria and the mitochondria of fungi and plants, the extra G of chicken mitochondrial tRNA(His) is not encoded in the gene . Therefore, the guanylate residue must be added post-transcriptionally, as demonstrated for the nuclear tRNA(His) in yeast and Drosophila . Analysis of a capped tRNA(His) precursor of chicken mitochondria suggests that addition of the extra G occurs independently of 3' end maturation . Since in the chicken mitochondrial tRNA(His) the extra G can be efficiently labeled by the capping assay, it should possess a 5'-terminal di- or triphosphate, which contrasts to the 5'-terminal monophosphate proposed for the nuclear encoded tRNA(His) . Our results imply that the ability of a mitochondrial RNA to be capped in vitro does not necessarily prove that it contains a transcription initiation site. FEBS Lett, 1990 Feb 12, 261(1), 14 - 8 Large scale preparation of homogeneous bacteriorhodopsin; Lorber B et al.; Homogeneous bacteriorhodopsin was obtained preparatively (100 mg batches) from purple membrane of Halobacterium halobium cells . The homogeneity of the protein was considerably affected by variations in the growth conditions of the bacteria . Fully matured bacteriorhodopsin having a blocked N-terminus and a homogeneous C-terminus, was reproducibly obtained when cells were grown in a sufficiently aerated medium. J Theor Biol, 1990 Feb 9, 142(3), 317 - 40 Verification of immune response optimality through cybernetic modeling; Batt BC et al.; An immune response cascade that is T cell independent begins with the stimulation of virgin lymphocytes by antigen to differentiate into large lymphocytes . These immune cells can either replicate themselves or differentiate into plasma cells or memory cells . Plasma cells produce antibody at a specific rate up to two orders of magnitude greater than large lymphocytes . However, plasma cells have short life-spans and cannot replicate . Memory cells produce only surface antibody, but in the event of a subsequent infection by the same antigen, memory cells revert rapidly to large lymphocytes . Immunologic memory is maintained throughout the organism's lifetime . Many immunologists believe that the optimal response strategy calls for large lymphocytes to replicate first, then differentiate into plasma cells and when the antigen has been nearly eliminated, they form memory cells . A mathematical model incorporating the concept of cybernetics has been developed to study the optimality of the immune response . Derived from the matching law of microeconomics, cybernetic variables control the allocation of large lymphocytes to maximize the instantaneous antibody production rate at any time during the response in order to most efficiently inactivate the antigen . A mouse is selected as the model organism and bacteria as the replicating antigen . In addition to verifying the optimal switching strategy, results showing how the immune response is affected by antigen growth rate, initial antigen concentration, and the number of antibodies required to eliminate an antigen are included. Am Surg, 1990 Feb, 56(2), 104 - 7 A prospective, randomized clinical trial of wound debridement versus conservative wound care in soft-tissue injury from civilian gunshot wounds; Brunner RG et al.; While clear-cut evidence exists documenting the extensive tissue destruction from blast and cavitation in high-velocity projectile injury and that wounds from projectiles of all velocities can be contaminated by bacteria potentially leading to infection, less is known about the tissue effects of lower-velocity projectile injury from gunshot wounds seen in the civilian sector . Despite this, traditional recommendations have supported debridement, admission, and aggressive wound care in these patients . This study will determine the effect of two methods of wound care on the outcome of soft-tissue gunshot wounds . Patients who had suffered a gunshot wound and were transported to the Trauma Center at the University of Florida Health Science Center postinjury were considered eligible for this study . All patients with torso injury, skeletal injury, neurovascular injury, or vascular proximity were excluded . Patients were then randomized to two treatment regimens based on a previously determined scheme . All wound care, follow-up, and healing evaluations were performed by the same individual . A total of 163 patients met the study criteria and they were randomized to the two treatment regimens: 89 patients, debridement and wound care; 74 patients, wound care alone . Each group was similar in age, sex, time to treatment, and caliber/velocity . Patients available for follow-up were similar in each group . Four patients in the wound-debridement group and two patients in the conservative wound-care group developed superficial infections . All infections responded to prompt local therapy . This study supports the conservative treatment of soft-tissue injury from low-velocity gunshot wounds. Am J Physiol, 1990 Feb, 258(2 Pt 1), L19 - 24 On the nature of enduring modifications induced in cells and organisms; Rubin H; The clarity of Mendelian genetics and the elegance of the molecular mechanisms of replication and readout of DNA have tended to obscure a solid body of evidence demonstrating that nongenetic, enduring modifications can be induced in the behavior of cells, modifications that continue to be expressed for many divisions after withdrawal of the inducing stimulus . The most prosaic case is the differentiated state of metazoan cells, which persists throughout the lifetime of the organism . Much less widely known but well-characterized examples are also cited for bacteria, protozoa, and cultured cells of higher plants and animals . The spontaneous neoplastic transformation of cultured mouse NIH 3T3 cells is introduced as an enduring adaptive response to moderate growth constraints . Evidence in support of the thesis that physiological adaptation is the driving force for chemically induced carcinogenesis in the intact animal is also presented . The cases described here involve integrated responses of many if not all of the regulatory components of the cell, rather than singular molecular mechanisms . The continuous generation of phenotypic heterogeneity, a process observed readily in cell culture, provides the basis for a model that accounts for enduring modifications . This model, designated progressive state selection, makes no attempt at a detailed biochemical explanation of heterogeneity, but uses it as a fundamental postulate to represent the adaptive behavior of the cells. J Cell Biol, 1990 Feb, 110(2), 367 - 78 Structure and function of the cytoskeleton of a Dictyostelium myosin-defective mutant; Fukui Y et al.; To study the role of conventional myosin in nonmuscle cells, we determined the cytoskeletal organization and physiological responses of a Dictyostelium myosin-defective mutant . Dictyostelium hmm cells were created by insertional mutagenesis of the myosin heavy chain gene (De Lozanne, A., and J . A . Spudich . 1987 . Science (Wash . DC) . 236: 1086-1091) . Western blot analysis using different mAbs confirms that hmm cells express a truncated myosin fragment of 140 kD (HMM-140 protein) instead of the normal 243-kD myosin heavy chain (MHC) . Spontaneous revertants appear at a frequency less than 4 x 10(-5), which synthesize normal myosin and are capable of forming thick filaments . In hmm cells, the HMM-140 protein is diffusely distributed in the cytoplasm, indicating that it cannot assemble into thick filaments . The actin distribution in these mutant cells appears similar to that of wild-type cells . However, there is a significant abnormality in the organization of cytoplasmic microtubules, which penetrate into lamellipodial regions . The microtubule networks consist of approximately 13 microtubules on average and their pattern is abnormal . Although hmm cells can form mitotic spindles, mitosis is not coordinated with normal furrow formation . The hmm cells are clearly defective in the contractile events that lead to normal cytokinesis . The retraction of different regions of the cell can result in the occasional pinching off of part of the cell . This process is not coupled with formation of mitotic spindles . There is no specific accumulation of HMM-140 in such constrictions, whereas 73% of such cells show actin concentrated in these regions . The mutant hmm cells are also deficient in capping of Con-A-bound surface receptors, but instead internalize this complex into the cytoplasm . The hmm cells display active phagocytosis of bacteria . Whereas actin is concentrated in the phagocytic cups, HMM-140 protein is not localized in these regions . cAMP, a chemoattractant that induces drastic rounding up and formation of surface blebs in wild type cells, does not induce rounding up in the hmm cells . A Triton-permeabilized cell model of the wild-type amebae contracts on reactivation with Mg-ATP, whereas a model of the hmm cell shows no detectable contraction . Our data demonstrate that the conventional myosin participates in the significant cortical motile activities of Dictyostelium cells, which include rounding up, constriction of cleavage furrows, capping surface receptors, and establishing cell polarity. J Endod, 1990 Feb, 16(2), 70 - 7 Clinical considerations of microleakage; Pashley DH; Most dental materials permit the microleakage of bacteria and bacterial products from oral fluids to reach dentin . Thus, an understanding of the clinical consequences of microleakage demands that we analyze the permeability characteristics of dentin . The more dentin surface that is exposed during tooth preparation, the greater the potential for microleakage . Thicker dentin is less permeable than thin dentin . Dentin over pulp horns is more permeable than central dentin . Similarly, the dentin making up axial walls is more permeable than dentin forming the pulpal floor of cavities . Coronal dentin is much more permeable than root dentin . In addition to the number of tubules per unit area, tubule diameter, and the location of the dentin in a tooth, another extremely important variable is the presence or absence of a smear layer . When smear layers are created, grinding debris are forced into each tubule to form a smear plug . Smear plugs are usually 1- to 2-microns long but may be 10-microns long . They are much longer than the smear layer thickness and they reduce dentin permeability more than the overlying smear layer . Smear layers constitute a natural cavity liner that reduces the permeability of dentin far more than any cavity varnish . However, its presence limits the strength of dentin bonding agents because of the relatively low cohesive forces holding the smear layer together and to dentin . If it is removed, the bond strengths of dentin adhesive resins increases, but removal also increases the potential liability for pulpal inflammation if the bond is not uniformly perfect.(ABSTRACT TRUNCATED AT 250 WORDS) G Ital Dermatol Venereol, 1990 Feb, 125(1-2), 7 - 13 {Clinico-pathogenetic observations on the subject of "superficial vasculitis"}; Petruzzellis V et al.; A clinical, histopathologic, laboratory and therapeutic study in 44 subjects with "superficial vasculitis", consecutively observed during the last two years, has been carried out . The findings showed that 1 . the anamnesis was sometimes not relevant; 2 . the clinical pictures were not in a regular manner correlated with histologic reports (a polymorphonuclear infiltrate with leukocytoclasis or a lymphomononuclear + one); and 3 . a non-correlation between clinical, histologic and etiologic data . Infectious agents were responsible, mostly pyogenic bacteria, Mycobacterium tuberculosis, and hepatitis B virus; Chlamydiae were rarely responsible, whereas the etiologic role of Toxoplasma was uncertain . In this respect, drugs were not relevant . All these agents seem to act on the immunological response of the patient with previous microvascular changes (diabetes, chronic venous insufficiency) . The latter condition seems to play a predominant role: the recovery of the lesions and/or their relapse shows the same behaviour both in case of etiological and symptomatic therapy. Vet Parasitol, 1990 Feb, 35(1-2), 71 - 7 Concurrent infection of enterocytes with Eimeria scabra and other enteropathogens in swine; Koudela B et al.; Bacteria were detected in the enterocytes of the distal jejunum in weaned pigs on Days 7 and 9 post-infection (DPI) with Eimeria scabra in addition to the developmental stages of the coccidia . Short rod-shaped bacteria were identified in approximately 60% of the enterocytes that contained developmental stages of E . scabra . No such bacteria were observed in cells where coccidia were absent . Gamonts of cryptosporidia were also observed within the microvillous zone of the enterocytes in the distal jejunum of weaned pigs on DPI 9 with E . scabra . Cryptosporidia were present only in enterocytes harbouring stages of E . scabra . Chlamydial particles were also found in the cytoplasm of enterocytes 7 DPI with E . scabra . The presence of other enteropathogens exclusively in the enterocytes containing developmental stages of coccidia suggests that the coccidium E . scabra facilitates the invasion and development of bacteria, cryptosporidia and chlamydia in the enterocytes. Xenobiotica, 1990 Feb, 20(2), 159 - 68 The stereoselectivity of 1,2-phenylethanediol and mandelic acid metabolism and disposition in the rat; Drummond L et al.; 1 . The steps involved in determining the chirality of the mandelic acid excreted by rats after administration of ethylbenzene and styrene were investigated by studying the fate of racemic, (R)- and (s)1,2-phenylethanediol, a precursor of mandelic acid . These investigations indicate the occurrence of two alternative routes of metabolism for 1,2-phenylethanediol, one involving retention of configuration and the other resulting in the loss of the chiral centre . 2 . The stereoselectivity of the disposition of mandelic acid was investigated; rats were dosed with mandelic acid either as the racemate or as the individual enantiomers, G.1.c.-mass spectrometry and h.p.l.c . were used to determine the enantiomers of mandelic acid . 3 . There were at least two routes by which mandelic acid could be metabolized and/or excreted; there is a stereoselective pathway in rat for (s)-mandelic acid, which gives rise to phenylglyoxylic acid . 4 . The chiral inversion of (s)-mandelic acid to (R)-mandelic acid is reported; although this has been observed in bacteria it has not previously been observed in mammals . 5 . The extent to which mandelic acid is metabolized to phenylglyoxylic acid is dependent on the enantiomeric composition of the mandelic acid administered . There is no evidence to indicate significant ketone-alcohol conversion, that is phenylglyoxylic acid is not significantly reduced to mandelic acid in vivo. J Biomed Mater Res, 1990 Feb, 24(2), 217 - 26 A new porous polyetherurethane wound covering; Bruin P et al.; A polyetherurethane (PEU) wound covering with non-interconnected micropores up to approximately 5 microns has been prepared by means of a phase inversion process . This highly elastic, very thin (15-20 microns), pliable wound covering showed good, immediate adherence to wet wound surfaces and high water vapor permeability, but was impermeable to bacteria . In guinea pigs epidermal wound healing of partial-thickness wounds under PEU wound coverings was accelerated compared with uncovered controls and an occlusive wound covering, OpSite . Water in liquid form or wound exudate could not leak through the PEU covering, but its high water vapor permeability induced concentration of the wound exudate into a jellylike clot layer, which apparently accelerated reepithelialization . The main conclusion from a clinical study on 20 donor sites was that the use of the PEU covering reduced pain, besides prevention of fluid retention . No differences in epithelialization were seen in comparison to tulle gras-treated wounds. Histochem J, 1990 Feb, 22(2), 102 - 5 Influence of colonizing micro-flora on the mucin histochemistry of the neonatal mouse colon; Hill RR et al.; Mucin histochemistry on sections of colon from germ-free and conventional mouse pups showed that all goblet cell mucins were sulphated at birth . During the first two weeks of post natal development, the pattern of mucin production in the ascending colon changed to a distribution of non-sulphated mucins towards the apical zone of the crypts and sulphated sialomucins basally . In conventional animals during the third postnatal week when the complex micro-flora of the colon was becoming established, the typical adult mucin distribution pattern developed, with sulphated mucins now confined to the upper third of the crypt . However, in the absence of a colonizing micro-flora crypt mucins become more and more sulphated until at weaning, most goblet cells of the ascending colon were producing fully or partially sulphated mucins, except for one or two cells at the very base of the crypt. J Lipid Res, 1990 Feb, 31(2), 289 - 98 Physicochemical and biological properties of natural and synthetic C-22 and C-23 hydroxylated bile acids; Roda A et al.; In order to define the effect of a side chain hydroxy group on bile acid (BA) physicochemical and biological properties, 23-hydroxylated bile acids were synthesized following a new efficient route involving the alpha-oxygenation of silylalkenes . 22-Hydroxylated bile acids were also studied . The synthesized bile acids included R and S epimers of 3 alpha,7 alpha,23-trihydroxy-5 beta-cholan-24-oic acid (23R epimer: phocaecholic acid), 3 alpha,12 alpha,23-trihydroxy-5 beta-cholan-24-oic (23R epimer: bitocholic acid), and 3 alpha,7 beta,23-trihydroxy-5 beta-cholan-24-oic acid . A 3 alpha,7 alpha,22-trihydroxy-5 beta-cholan-24-oic acid (haemulcholic acid) was also studied . The presence of a hydroxy group on the side chain slightly modified the physicochemical behavior in aqueous solution with respect to common BA: the critical micellar concentration (CMC) and the hydrophilicity were similar to naturally occurring trihydroxy BA such as cholic acid . The pKa value was lowered by 1.5 units with respect to common BA, being 3.8 for all the C-23 hydroxy BA . C-22 had a higher pKa (4.2) as a result of the increased distance of the hydroxy group from the carboxy group . When the C-23 hydroxylated BA were intravenously administered to bile fistula rats, they were efficiently recovered in bile (more than 80% unmodified) while the corresponding analogs, lacking the 23- hydroxy group, were almost completely glycine- or taurine-conjugated . On the other hand, the C-22 hydroxylated BA were extensively conjugated with taurine and less than 40% of the administered dose was secreted without being conjugated . In the presence of intestinal bacteria, they were mostly metabolized to the corresponding 7-dehydroxylated compound similar to common BA with the exception of bitocholic acid which was relatively stable . The presence of a hydroxy group at the C-23 position increased the acidity of the BA and this accounted for poor absorption within the biliary tree and efficient biliary secretion without the need for conjugation . 3 alpha,7 beta-23 R/S trihydroxy-5 beta-cholan-24-oic acids could improve the efficiency of ursodeoxycholic acid (UDCA) for gallstone dissolution or cholestatic syndrome therapy, as it is relatively hydrophilic and efficiently secreted into bile without altering the glycine and taurine hepatic pool. J Anim Sci, 1990 Feb, 68(2), 467 - 77 Intestinal supply of amino acids in sheep fed alkaline hydrogen peroxide-treated wheat straw-based diets supplemented with soybean meal or combinations of corn gluten meal and blood meal; Cecava MJ et al.; The intestinal supply of amino acids (AA) in sheep fed alkaline hydrogen peroxide-treated wheat straw (AHPWS)-based diets supplemented with soybean meal (SBM) or corn grain plus combinations of corn gluten meal (CGM) and blood meal (BM) was measured in a 5 X 5 latin square . Sheep (avg wt 45 kg) with ruminal, duodenal and ileal cannulas were fed diets containing 65% AHPWS supplemented with the following protein sources: soybean meal (SBM), corn gluten meal (CGM), blood meal (BM), 2/3 CGM:1/3 BM and 1/3 CGM:2/3 BM . Total nitrogen (N) flow at the duodenum was not affected (P greater than .05) by protein source . Flows of bacterial N and AA increased (P less than .05) and flows of nonbacterial N and AA decreased (P less than .05) when wethers were fed SBM vs corn plus other protein sources . When diets contained SBM, quantities of total AA at the duodenum were lower (P less than .05) and the profile of AA supplied to the intestine was altered substantially . Total flows of AA at the duodenum and total quantities of AA disappearing from the small intestine were similar (P greater than .05) for all diets containing BM and CGM, but flows and disappearance of valine, histidine, lysine and arginine increased linearly (P less than .05), whereas flows and disappearance of leucine, isoleucine and methionine decreased linearly (P less than .05) as BM replaced CGM in the diets . Results suggest that quantities of individual AA flowing to the duodenum and disappearing from the intestine of wethers fed AHPWS-based diets can be altered by source of dietary protein . Furthermore, feeding protein sources resistant to ruminal degradation in combination may improve the profile of AA supplied to the intestine. Ecotoxicol Environ Saf, 1990 Feb, 19(1), 24 - 32 The effects of salinity and temperature on the toxicity of copper to 1-day and 7-day-old larvae of Hediste (Nereis) diversicolor (O . F . Muller); Ozoh PT et al.; The effects of salinity and temperature on the toxicity of copper to 1-day and 7-day-old larvae of Hediste diversicolor were tested using factorial analysis . Mortality of larvae was influenced by salinity, temperature, and copper, but 1-day-old larvae were more susceptible than the 7-day-old larvae . Resistance to copper increased with age . Low concentrations of 5 and 20 micrograms/liter copper were more inhibitory to 1-day and 7-day old larvae, respectively, than were higher copper levels . The responses of 1-day-old larvae to copper levels from 0 to 20 micrograms/liter and 7-day-old to copper levels from 0 to 100 micrograms/liter copper were complex . Increasing salinity 7.6 to 30.5% reduced both copper toxicity and protozoa/bacteria attacks. Immunology, 1990 Feb, 69(2), 190 - 4 The serum polymeric IgA antibody response to typhoid vaccination; its relationship to the intestinal IgA response; Bartholomeusz RC et al.; The relationship between the IgA antibody response in serum (total and polymeric IgA) and intestinal secretions was examined in volunteers subjected to oral and parenteral typhoid vaccination . After oral vaccination (three doses of 10(11) live Ty21a vaccine given at 48-hr intervals), serum pIgA antibody to typhoid lipopolysaccharide (LPS) was detected in seven of the 14 subjects (46.4 +/- 59 U/100 microliters, mean +/- SD) . However, all 14 showed a significant intestinal IgA response (993 +/- 2516 and 9349 +/- 6754 U/mg pre- and post-vaccine; t = 5.25, P = 0.0002) . The level of pIgA antibody declined rapidly, whereas intestinal IgA antibody levels remained elevated . Serum pIgA antibody was also found after parenteral immunization (two doses of 5 X 10(8) heat-killed bacteria given 14 days apart to six subjects), but an intestinal IgA antibody response was detected in these individuals only after a subsequent course of the oral vaccine given 1 month after initial parenteral immunization . Changes in serum pIgA antibody followed those of total serum IgA antibody rather than those of intestinal antibody . The results indicate that a serum pIgA response can be induced by an antigenic stimulus delivered either orally or parenterally, whereas an intestinal IgA response is induced only by a local antigen stimulus . The regulation of serum pIgA and intestinal IgA appear to be independent. Cancer Lett, 1990 Feb, 49(2), 89 - 98 Carcinogenicity tests of fecapentaene-12 in mice and rats; Weisburger JH et al.; Fecapentaenes, a class of direct-acting bacterial mutagens, have been isolated from the feces and intestinal tract of humans on a Western meat-containing diet . Two bioassays to test pure fecapentaene-12 (FP-12) for carcinogenicity were performed . FP-12 in dimethylsulfoxide (DMSO) solution was injected i.p . into newborn ICR/MA mice on days 1, 3, 7, 10, 14 and 21 . The mice killed after 21 months had neoplasms in liver, lung, glandular stomach and subcutaneous fibrosarcoma . Intrarectal (i.r.) infusion of FP-12 in an aqueous vehicle into male F344 rats for 71 weeks, and killing the rats after 21 weeks more, displayed no evidence of neoplasia associated with FP-12 exposure . The positive control, N-nitrosomethylurea (NMU), given i.r . as 4 2-mg doses in 2 weeks, as expected, yielded multiple colonic neoplasms in less than 11 months . Fecapentaene may exert its effect in bacteria and in newborn mice through the generation of hydroxy radicals . However, adult rodent and human colon may have adequate biochemical defense mechanisms against low level, even continuous exposures to chemicals like FP-12, and thus be at low risk of neoplasia, as was found. Arch Biochem Biophys, 1990 Feb 1, 276(2), 451 - 9 Precursor-product relationship of larger to smaller molecular forms of the BAL 31 nuclease from Alteromonas espejiana: preferential removal of duplex exonuclease relative to endonuclease activity by proteolysis; Hauser CR et al.; Two molecularly and kinetically distinct major species of the extracellular nuclease BAL 31 from Alteromonas espejiana, previously characterized as the "fast" (F) and "slow" (S) BAL 31 nucleases, have been evidenced to derive from proteolysis starting from a still larger (approximately 120 kDa) precursor nuclease . The expected protease activity in the culture fluid has been confirmed and is strongly dependent on the cell growth phase . The disappearance of the largest nuclease species with the concomitant sequential appearance of first the F and then the S species has been demonstrated for nuclease obtained from culture supernatants as a function of cell growth phase . Nuclease from periplasmic extracts displayed very little of the F and S nucleases . Treatment of purified F nuclease with Pronase or subtilisin readily converted it to species with only a few percent of the native exonuclease activity against duplex DNA but retaining much of the initial activity against single-stranded DNA . Electrophoresis in nuclease-detecting gels demonstrated a parallel conversion of the larger species to one indistinguishable in molecular weight from the S species . The observed loss of exonuclease activity could correspond to the conversion of the F to the S nuclease . However, treatment of S nuclease with subtilisin resulted in a drastic reduction of exonuclease activity of this enzyme on duplex DNA with retention of most of the activity against single-stranded and nicked circular duplex DNA substrates . Evidence of internal proteolysis of the S nuclease could be seen after electrophoresis in denaturing gels but only after the denaturation buffer was adjusted to 6 M in urea . The preferential removal of the exonuclease activity may enhance the usefulness of the BAL 31 nuclease in such applications as heteroduplex mapping. Am Surg, 1990 Feb, 56(2), 96 - 9 Is diagnostic peritoneal lavage for blunt trauma obsolete? Hawkins ML, Bailey RL Jr, Carraway RP. Diagnostic peritoneal lavage was 97 percent accurate, with a 2 percent false positive rate and a 1 percent false negative rate in this series of 414 patients . The ease, safety, and accuracy of diagnostic peritoneal lavage justify its continued use in evaluating these patients . Recent studies show computerized tomography (CT) can be highly accurate in detecting intra-abdominal injuries after blunt trauma . We reviewed our experience with diagnostic peritoneal lavage (DPL) to evaluate whether the accuracy, safety, speed, and cost justified its continued use . Four hundred fifteen DPLs were performed on 414 patients from February 1, 1983, through December 31, 1987 . All DPLs were done by the open technique . The lavage was considered grossly positive if 10 cc gross blood were aspirated . If there were greater than 100,000 red blood cells (RBC)/mm3, greater than 500 white blood cells (WBC)/mm3, elevated amylase or bilirubin, or bacteria or vegetable fibers the lavage was microscopically positive . There were no cases with elevated bilirubin, amylase, or presence of bacteria . All four cases with "rare vegetable fibers" were false positive . Six DPLs were for penetrating trauma to the lower chest or back . There were 291 negative lavages, including five false negatives (1%), and 124 positive DPLs, including seven false positives (2%), resulting in a crude accuracy of 97 percent . Three of the five false negative lavages had a ruptured diaphragm as the only intra-abdominal injury . There was one minor complication . DPL was usually performed in the trauma resuscitation room during the secondary survey . At our institution, the total fees for DPL are +185 less than the fees for CT . DPL is accurate, rapid, safe, and avoids the disruption of patient care that results in the radiology suite . DPL remains our procedure of choice for evaluating blunt abdominal trauma in the adult. J Leukoc Biol, 1990 Feb, 47(2), 135 - 41 Natural killer cell activity and macrophage-dependent inhibition of growth or killing of Mycobacterium avium complex in a mouse model; Bermudez LE et al.; Natural killer (NK) cells from spleens of normal and Mycobacterium avium complex (MAC)-infected C57 black mice (C57 BL/6 bg/+) were examined for their capacity to activate splenic and peritoneal macrophages from beige mice to inhibit or kill intracellular MAC . Peritoneal and splenic macrophages from beige mice were exposed in vitro to NK cells obtained from MAC-infected and uninfected black mice . NK cells from uninfected black mice were also treated in vitro with recombinant interleukin-2 (IL-2) for 48 h before incubation with macrophages . While control macrophages supported intracellular growth of MAC, macrophages exposed to unactivated NK cells inhibited growth of the intracellular bacteria, as determined 4 days after infection . IL-2 stimulated NK cells, and NK cells obtained from MAC-infected animals were able to activate murine macrophages in vitro to inhibit growth or kill 40.0 +/- 5% and 61.3 +/- 6% of the intracellular bacteria, respectively . In other experiments, beige mice (C57 BL/6 bg/bg) were treated intraperitoneally with NK cells obtained from MAC-infected and uninfected C57 black mice . Peritoneal macrophages harvested from beige mice treated with NK cells activated in vitro with IL-2 killed 24.4 +/- 4% of intracellular bacteria by day 4 after infection . Macrophages obtained from animals treated with NK cells harvested from MAC-infected black mice killed 58.8 +/- 7% of intracellular bacteria by 4 days after infection, in contrast with intracellular growth observed in macrophages obtained from untreated animals and from animals treated with Hanks' solution or unactivated NK cells . These crossover studies suggest that NK cells may be important in host defense against MAC. J Bacteriol, 1990 Feb, 172(2), 1142 - 4 VirA, a coregulator of Ti-specified virulence genes, is phosphorylated in vitro; Huang Y et al.; High-level expression of a chimeric virA gene was obtained by replacing the first 524 codons of virA with the first half of trpE . The encoded fusion protein was isolated and found to exhibit autokinase activity . Therefore, a kinase domain is in the C-terminal portion of VirA, and protein phosphorylation may be an important feature of VirA function. J Bacteriol, 1990 Feb, 172(2), 1070 - 6 Pyrogallol-to-phloroglucinol conversion and other hydroxyl-transfer reactions catalyzed by cell extracts of Pelobacter acidigallici; Brune A et al.; Permeabilized cells and cell extracts of Pelobacter acidigallici catalyzed the conversion of pyrogallol (1,2,3-trihydroxybenzene) to phloroglucinol (1,3,5-trihydroxybenzene) in the presence of 1,2,3,5-tetrahydroxybenzene . Pyrogallol consumption by resting cells stopped after lysis by French press or mild detergent (cetyltrimethylammonium bromide {CTAB}) treatment . Addition of 1,2,3,5-tetrahydroxybenzene to the assay mixture restored pyrogallol consumption and led to stoichiometric phloroglucinol accumulation . The stoichiometry of pyrogallol conversion to phloroglucinol was independent of the amount of tetrahydroxybenzene added . The tetrahydroxybenzene concentration limited the velocity of the transhydroxylation reaction, which reached a maximum at 1.5 mM tetrahydroxybenzene (1 U/mg of protein) . Transhydroxylation was shown to be reversible . The equilibrium constant of the reaction was determined, and the free-energy change (delta G degree') of phloroglucinol formation from pyrogallol was calculated to be -15.5 kJ/mol . Permeabilized cells and cell extracts also catalyzed the transfer of hydroxyl moieties between other hydroxylated benzenes . Tetrahydroxybenzene and hydroxyhydroquinone participated as hydroxyl donors and as hydroxyl acceptors in the reaction, whereas pyrogallol, resorcinol, and phloroglucinol were hydroxylated by both donors . A novel mechanism deduced from these data involves intermolecular transfer of the hydroxyl moiety from the cosubstrate (1,2,3,5-tetrahydroxybenzene) to the substrate (pyrogallol), thus forming the product (phloroglucinol) and regenerating the cosubstrate. Infect Immun, 1990 Feb, 58(2), 489 - 94 Antibody response to Brucella ovis outer membrane proteins in ovine brucellosis; Riezu-Boj JI et al.; Hot saline extracts of Brucella ovis were composed of vesicles with outer membrane proteins (OMPs), lipopolysaccharide, and phospholipid as constituents . Extraction with petroleum ether-chloroform-phenol yielded a protein fraction free of detectable lipopolysaccharide, in which group 3 OMPs (28,500 apparent molecular weight {28.5K}, 27.0K, and 25.5K) represented 81% of the total . Group 1 OMPs and 67.0K, 22.5K to 21.5K, and 19.5K to 18.0K proteins were also detected . Adsorption of immune sera with whole bacteria suggested that group 3 OMPs and 67.0K, 22.5K to 21.5K, and 19.5K to 18.0K proteins had antigenic determinants exposed on the surfaces of both B . ovis and rough B . melitensis cells but not on smooth B . melitensis cells . Antibodies to group 3 OMPs and the 67.0K protein in the sera of 93 and 87%, respectively, of B . ovis-infected rams were found by immunoblotting . Antibodies to other proteins were present in 67% of these animals . Compared with B . ovis-infected rams which had not developed lesions, rams with epididymo-orchitis had antibodies to a larger variety of proteins . Although ewes infected with B . melitensis also showed antibodies to OMPs, the immunoblot reactions were less intense. Bioessays, 1990 Feb, 12(2), 74 - 9 The regulation of DNA repair during development; Mitchell DL et al.; DNA repair is important in such phenomena as carcinogenesis and aging . While much is known about DNA repair in single-cell systems such as bacteria, yeast, and cultured mammalian cells, it is necessary to examine DNA repair in a developmental context in order to completely understand its processes in complex metazoa such as man . We present data to support the notion that proliferating cells from organ systems, tumors, and embryos have a greater DNA repair capacity than terminally differentiated, nonproliferating cells . Differential expression of repair genes and accessibility of chromatin to repair enzymes are considered as determinants in the developmental regulation of DNA repair. Sports Med, 1990 Feb, 9(2), 100 - 19 Common cutaneous disorders in athletes; Conklin RJ; Athletic activity may cause or aggravate skin disorders, which in turn may diminish athletic performance . Since many sporting activities necessitate prolonged exposure to the sun, athletes must avoid painful sunburn which will adversely affect their performance . Drugs and chemicals also may cause photoallergic and/or phototoxic reactions, including polymorphous light eruption and athletes should thus avoid photosensitising drugs and chemicals . The effects of chronic ultraviolet exposure include ageing, pigmentation and skin cancers . The most effective protection against excessive exposure to sunlight is the use of sunscreens, although inadequate application and poor protection in the UVA spectrum may diminish their effectiveness and contact allergies may create other problems . Viral, bacterial and fungal infections are common in athletes due to heat, friction and contact with others . Herpes simplex may be treated with any drying agents (e.g . alcohol) as they are as effective as more expensive topical agents such as acyclovir . Molluscum contagiosum may be spread by close contact or water contact and is treated by superficial incision, cryotherapy or standard wart varnishes . Plantar wart infection is transmitted by swimming pool decks, changing rooms and hand-to-hand from weights in gymnasiums . Plantar warts presenting with pain may be aggressively treated, by blunt dissection, but painless ones are best treated conservatively . Impetigo and folliculitis often develop after trauma . Antibiotics are effective against mild infections while abrasions and lacerations should be cleansed and dressed with occlusive dressings . Diphtheroid bacteria in moist footwear may produce pitted keratolysis and erythrasma . Tinea pedis is common in athletes and probably originates in swimming pools, gymnasium floors and locker rooms . Interdigital, dry-moccasin and pustular-midsole forms can be distinguished . The latter two forms respond to topical antifungal agents, while the interdigital form, a mixed fungal/bacterial infection, is treated with debridement, antibiotics and drying routine similar to the therapy of otitis externa . Nail infections by a variety of organisms may appear as onycholysis with or without paronychia and should be treated with the appropriate antibiotics . Tinea versicolor occurs in heat and humidity . Since Pityrosporum orbiculare is part of the normal flora it often recurs, necessitating regular treatment . Acute trauma injuries include contusions, black heel or petichiae of the heel, black toe (bleeding under the nail), 'jogger's nipple' caused by chafing, and foot blisters . Chronic trauma may result in calluses, corns and paronychia . Plantar corns can be disabling and may be caused by overly tight shoes or abnormalities in biomechanics; treatment includes restoring normal foot function and minimal surgical procedures . Paronychia is treated best by wedge resection.(ABSTRACT TRUNCATED AT 400 WORDS) Proc Natl Acad Sci U S A, 1990 Feb, 87(4), 1273 - 7 Mutagenesis of Thr-286 in monomeric Ca2+/calmodulin-dependent protein kinase II eliminates Ca2+/calmodulin-independent activity; Waxham MN et al.; We have examined the role of Thr-286 autophosphorylation in the autoregulation of Ca2+/calmodulin-dependent protein kinase II . Using site-directed mutagenesis, we have substituted alanine or serine for Thr-286, or isoleucine for Arg-283, in the 50-kDa subunit of the kinase and expressed each protein in bacteria . Activation and autophosphorylation of all four enzymes were stringently dependent on Ca2+/calmodulin, indicating that neither Arg-283 nor Thr-286 is an absolute requirement for the pseudosubstrate inhibition of the enzyme . Autophosphorylation of the Ile-283 or Ala-286 enzyme generated little, if any, Ca2+/calmodulin-independent kinase activity, unlike the parent (Thr-286) or Ser-286 enzyme . The enzymes expressed in bacteria are predominantly monomeric, indicating that the generation of Ca2+/calmodulin-independent activity does not require the cooperative interactions of subunits normally present in the brain holoenzyme. Mutat Res, 1990 Feb, 240(2), 47 - 58 Tests for the genotoxicity of m-AMSA, etoposide, teniposide and ellipticine in Neurospora crassa; Gupta R; The antitumor agents m-AMSA, etoposide, teniposide and ellipticine have been reported to be potent clastogens in mammalian cells but non- or weakly mutagenic in bacteria; these observations have been correlated to the interference of these chemicals with DNA topoisomerase II activity in the former, but not in the latter, organisms . The genotoxicity of these 4 agents was evaluated using ad-3 reverse- and forward-mutation tests in Neurospora crassa . These agents (up to 0.8 mumole/plate) did not cause reversion in conidia of the ad-3A frameshift strains N24 and 12-9-26 using the overlay plate test, as contrasted to the positive control frameshift mutagen ICR-170 . Heterokaryon 12 (H-12) of N . crassa permits the recovery of all classes of forward mutation at the ad-3+ region, including multilocus deletions . Using resting conidia of H-12 in a suspension assay, ellipticine was moderately mutagenic but no increase in ad-3 mutants was noted with the other 3 agents at a dose of 100 micrograms/ml . In vegetative cultures of H-12 grown in the presence of these agents, all 4 agents were nonmutagenic at a dose of 100 micrograms/ml . The positive control mutagen ICR-170 was mutagenic in both resting conidia and growing cultures of H-12 . A similarity between the topoisomerase II of N . crassa and DNA gyrase of bacteria is suggested. Math Biosci, 1990 Feb, 98(1), 73 - 102 Analysis of the effects of immune cell motility and chemotaxis on target elimination dynamics; Fisher ES et al.; White blood cells of the immune system must encounter specific targets such as bacteria, malignant cells, virus-infected cells or other cells of the immune response in order to carry out their function of protecting the host from infectious and malignant disease . To analyze the dynamics of this process, a mathematical model has been developed for elimination of proliferating targets by a constant population of motile immune system cells in two dimensions . Encounter is assumed to be the rate-limiting step for elimination . This model makes use of a previously derived analysis of single cell-target encounter times, which yields an encounter rate constant that is incorporated into a kinetic conservation equation for target number density . This paper focuses on the influence of directed cell movement, or chemotaxis, as well as other cell motility properties, such as cell speed and persistence, on target elimination dynamics . A particularly significant result is that a given relative decrease in chemotactic responsiveness leads to much more severe deficiencies in target clearance rates for low levels of baseline chemotactic responsiveness than for high levels of baseline responsiveness . The general model results are then applied to the particular example of bacterial clearance from the lung surface by alveolar macrophages . It is shown that moderate levels of macrophage chemotactic responsiveness, similar to those measured in vitro, can account for the experimentally observed rates of bacterial elimination from the lung for typical values of bacterial specific growth rate and alveolar macrophage number density. Pneumologie, 1990 Feb, 44 Suppl 1, 455 - 7 {Differentiation of M . tuberculosis and M . avium complex using various monoclonal antibodies}; Wurche T et al.; M . avium-complex (MAC) is the cause of the most bacterial infections in AIDS patients . Because of the high resistance of MAC, a rapid differentiation between M . tuberculosis and MAC is of great interest . In an enzyme-linked immunosorbent assay (ELISA) we tested three monoclonal antibodies BS 103, BS 104, BS 113 and the combination of BS 103/BS 113, which bind selectively to the cell wall of M . tuberculosis . 98 MAC isolates from AIDS patients and 233 M . tuberculosis isolates from patients with lung disease were tested after primary culture of the bacteria on Lowenstein-Jensen media . ELISA could clearly discriminate MAC from M . tuberculosis with a specificity of 100% and a sensitivity of 98-100% . With these monoclonal antibodies the time for differentiation can be reduced by several weeks. Immunobiology, 1990 Feb, 180(2-3), 208 - 20 Protection of mice against Brucella abortus by immunization with polyclonal anti-idiotype antibodies; Beauclair KD et al.; Polyclonal goat anti-idiotypic antibodies containing internal images which mimic Brucella abortus antigens were generated from rabbit polyclonal idiotypes specific for partially purified extract of B . abortus (PX III) . The anti-idiotypic antibodies were purified using two-step immunoaffinity column chromatography . The presence of internal images was demonstrated by competitive inhibition analysis using an enzyme-linked immunosorbent assay . Several groups of BALB/c mice were vaccinated with the anti-idiotypic antibodies . The vaccinated mice showed a high serum titer of antibodies specific for B . abortus . When the vaccinated mice were challenged with a virulent B . abortus strain 2308, greater than 90% reduction of bacteria in the spleen as compared to the unvaccinated control groups was seen . Immunoblotting experiments using antiserum from vaccinated mice demonstrate the ability to distinguish vaccinated mice from B . abortus infected mice . Our data indicate that the anti-idiotypic antibody containing internal images of B . abortus may be used as a vaccine and the induced antibody can be distinguished by immunoblotting from antibodies generated by natural infection with B . abortus. Zentralbl Veterinarmed A, 1990 Feb, 37(1), 51 - 60 Influence of exogenous estradiol on the concentration of antitrypsin, albumin, N-acetyl-beta-D-glucosaminidase and somatic cells in milk of cows at various lactation stages; Saad AM et al.; Estradiol benzoate (0.02 mg/kg of body weight) was injected (i.m.) daily to 13 ovariectomized cows in their first or third lactations during early (60-90 days), mid (140-200 days) and late (240-300 days) lactation . The majority of the udder quarters were free of bacteria throughout the experiment . Signs of clinical mastitis were observed after 9-19 days of treatment in cows at mid and late lactation . This was accompanied by a decrease in milk yield, an increase in milk somatic cell count (MSCC) and increases in milk concentration of antitrypsin and serum albumin (as indicator of increased permeability); and milk NAGase activity (as indicator of epithelial cell damage or release of the enzyme from phagocytes) . NAGase activity was first to increase and on some occasions was not accompanied by an increased MSCC . The response was either absent (first lactation) or relatively mild (third lactation) in cows at the early stages of lactation . The intensity of the inflammatory response increased with advancing lactation stage . Generally, the response of cows in the third lactation was greater than that of cows in the first lactation at a corresponding stage of lactation . Three cows which received the control treatment with vehicle alone showed no changes in any of the parameters except a slight decrease in antitrypsin . All inflammatory indicators correlated negatively with quarter milk production; milk NAGase activity was superior in this respect (r = -0.75) . All correlation coefficients were generally higher after, rather than before the hormone treatment. Am J Surg, 1990 Feb, 159(2), 222 - 5 Three-phase study of phlebitis in patients receiving peripheral intravenous hyperalimentation; Rypins EB et al.; We found clinical phlebitis in 57 of 88 patients with peripheral hyperalimentation (65%) . To determine if this was a problem common to all intravenous fluid therapy at our hospital, we performed a point-prevalence study . The prevalence of phlebitis in nonhyperalimentation intravenous patients was 18% (84 of 456 patients) . We then performed a randomized, prospective, double-blind trial of sham versus standard in-line filters to determine if bacteria or filterable particulate matter was responsible for phlebitis in the peripheral hyperalimentation group . The standard-filter group had a phlebitis rate of 74% compared with 64% in the sham-filter group . We then eliminated in-line filters and replaced the standard glucose-based solution with a glycerol-based peripheral hyperalimentation solution . The phlebitis rate decreased from 68% to 27% (p less than 0.001) . In conclusion, phlebitis in peripheral hyperalimentation patients was probably due to chemical properties of the peripheral hyperalimentation solution rather than bacteria or particulates. Crit Care Med, 1990 Feb, 18(2 Suppl), S149 - 53 Nutritional immunomodulation in burn patients; Alexander JW et al.; A series of laboratory experiments in thermally injured guinea pigs has shown that feeding by the enteral route immediately after injury results in a decreased metabolic response by preventing loss of the GI barrier to the entrance of intestinal endotoxin and bacteria . Feeding by the iv route or giving crystalline amino acids instead of intact protein does not prevent atrophy of the intestine, nor does it prevent the hypermetabolic response . Optimal diets for nutritional support of burn patients contain 20% of energy from whey protein, 2% from arginine, 0.5% from cysteine, and 0.5% from histidine . Lipids comprise 15% of nonprotein calories with 50% fish oil (high in omega-3 fatty acids) and 50% safflower oil (high in linoleic acid) . In a prospective clinical study, administration of this new diet was found to reduce wound infection (p less than .03), shorten hospital stay (p less than .02), and reduce death (p less than .06) when compared to other standard enteral formulations. Jpn J Med Sci Biol, 1990 Feb, 43(1), 23 - 7 Evaluation of a simple staining method for flagella using "flagella staining solution-Shionogi"; Inoue K et al.; A flagella staining method using "Flagella Staining Solution-Shionogi" was tested for its reliability and convenience for a routine procedure in the clinical laboratory . The staining has proved to be accurate and reliable and can be used easily as a simple staining method. Teratology, 1990 Feb, 41(2), 161 - 6 Inhibitory effects of cobalt chloride and cinnamaldehyde on 5-azacytidine-induced digital malformations in rats; Kurishita A et al.; The effect of two DNA repair inhibitors in bacteria, cobalt chloride and cinnamaldehyde, on 5-azacytidine (5-AC)-induced digital malformations was studied . Both agents inhibited the induced digital malformations . The effect of cobalt chloride was significant 3 hr before to 1 hr after the 5-AC treatment, and the effect of cinnamaldehyde was significant 3 hr before to 24 hr after the treatment . However, an increase in fetal mortality was observed with the latter agent . The mechanisms underlying the suppressive effects of both agents may be different, but their natures require elucidation. Am Biotechnol Lab, 1990 Feb, 8(2), 32 - 4 Quinoproteins: a new class of enzymes with potential use as biosensors; Davidson VL; The field of the enzymology of quinoproteins is still in its infancy . As more progress is made toward understanding the mechanisms of catalysis and electron transfer by protein-bound PQQ, it will become possible to more logically design specific quinoprotein-based electrodes . The characterization of new quinoproteins is progressing at a rapid rate . As new quinoproteins are found, the potential range of applications for use of these enzymes in biosensors will increase as well. Philos Trans R Soc Lond B Biol Sci, 1990 Jan 30, 326(1236), 437 - 54 Resolution and reconstitution of anion exchange reactions; Maloney PC; To illustrate the emerging class of anion exchange proteins in bacteria, this article discusses the biochemical and physiological properties of phosphate (Pi)-linked antiporters that accept glucose 6-phosphate (G6P) as their primary substrate . These systems have a bifunctional active site that binds a pair of negative charges, whether presented as a single divalent anion or a pair of monovalent substrates . Exchange stoichiometry therefore moves between the limits of 2:1 and 2:2 according to the ratio of mono- and divalent substrates at either membrane surface . This predicts an interesting reaction sequence in vivo because internal pH is more alkaline than external pH; one expects an asymmetric exchange as a pair of monovalent G6P anions moves against a single divalent G6P, and in this way an otherwise futile self-exchange of G6P can result in a net inward flux driven (indirectly) by the pH gradient . Despite their biochemical complexity, at a molecular level the Pi-linked antiporters resemble other secondary carriers . Indeed, the current listing of nearly two dozen such proteins suggests a structural theme in which the minimal functional unit has two sets of six transmembrane alpha helices separated by a central hydrophilic loop . Presently described examples show that this topology can derive from either a single protein or from pairs of identical subunits . The finding of this common structure makes it possible to begin building more detailed structural models that have more general implications. Cell, 1990 Jan 26, 60(2), 199 - 209 Complex formed by complementary RNA stem-loops and its stabilization by a protein: function of CoIE1 Rom protein; Eguchi Y et al.; A small plasmid-specified RNA (RNA I) inhibits formation of the RNA primer for CoIE1 DNA replication by binding to its precursor (RNA II) . Binding is modulated by the plasmid-specified Rom protein . Both in the presence and absence of Rom, binding starts with interaction between loops of RNAs . To understand the mechanism of binding, we examined the interactions of pairs of single stem-loops that are complementary fragments of RNA I and RNA II . We found that these complementary single stem-loops bind to each other at their loops, forming an RNAase V1-sensitive structure . Rom protects the complex from cleavage and from alkylation of phosphate groups by ethyinitrosourea . A single dimer of Rom binds to the complex by recognizing the structure rather than its exact nucleotide sequence . Rom enhances complex formation by decreasing the rate of dissociation of the complex . Structures of RNA complexes formed in the presence and absence of Rom are proposed. J Biol Chem, 1990 Jan 25, 265(3), 1430 - 5 Rat ATP citrate-lyase . Molecular cloning and sequence analysis of a full-length cDNA and mRNA abundance as a function of diet, organ, and age; Elshourbagy NA et al.; ATP citrate-lyase is the primary enzyme responsible for the synthesis of cytosolic acetyl-CoA in many tissues . We have isolated a full-length cDNA copy of 4.3 kilobase pairs encoding the ATP-citrate lyase mRNA by screening rat liver cDNA library using oligonucleotide probes designed from peptide sequences obtained from the purified rat enzyme . Expression of this cDNA in bacteria, followed by immunoblotting with antibody directed against the ATP citrate-lyase, further demonstrated the identity of this clone . Nucleic acid sequence data indicate that the cDNA contains the complete coding region for the enzyme, which is 1100 amino acids in length with a calculated molecular weight of 121,293 . RNA blot analysis indicated an mRNA species of about 4.3 kilobase pairs in livers of chow-fed rats . Rats maintained on low fat, high carbohydrate diets exhibited a striking increase (50-fold) in the level of liver ATP citrate-lyase mRNA as compared with the control animals maintained on a normal diet . The tissue distribution of this mRNA in chow-fed animals revealed a relatively high abundance of the message in liver and adrenal, moderate levels were found in lung, brain, and large intestine with only trace amounts of the message in small intestine, stomach, testis, spleen, pancreas, kidney, and heart . During rat development, the ATP citrate-lyase mRNA was relatively high in the liver at parturition, followed by a reduction in its level during suckling . Higher amounts of the mRNA were detected again in adult animals . The isolation and characterization of the mRNA for ATP citrate-lyase will allow further studies on the reaction mechanism and metabolic regulation of this key enzyme in lipogenesis and cholesterogenesis. J Biol Chem, 1990 Jan 25, 265(3), 1496 - 501 Enzymic properties of thermopsin; Fusek M et al.; The specificity of thermopsin, a thermostable acid protease from Sulfolobus acidocaldarius, was studied using oxidized insulin B chain as substrate followed by peptide isolation and identification . The following bonds were hydrolyzed: Leu-Val, Leu-Tyr, Phe-Phe, Phe-Tyr, and Tyr-Thr . Thus, the specificity of ther