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Adherence and internalization of Streptococcus uberis to bovine mammary epithelial cells are mediated by host cell proteoglycans.

The role of cell glycosaminoglycans (GAG) on adherence and internalization of Streptococcus uberis to bovine mammary epithelial cells was evaluated by adherence/internalization competition assays, by removal of GAG from the host cell surface and by inhibition of GAG glycosylation in the host cell. Heparin (HEP), heparan sulfate (HSA), chondroitin sulfate A (CSA) and chondroitin sulfate B (CSB) inhibited adherence and internalization of S. uberis in a dose-dependent manner. However, inhibition was lower with CSA and CSB than that observed with HEP and HSA. Adherence and internalization were also inhibited upon treatment of mammary epithelial cells with GAG lyases. The greatest inhibition was observed with heparinase I. Tunicamycin, an inhibitor of mammalian cell glycosylation of cell surface glycoproteins, markedly inhibited internalization of S. uberis into mammary epithelial cells. Differences between strains were observed. These results suggest that a HSA proteoglycan receptor on the host cell surface may mediate S. uberis adherence to and internalization of bovine mammary epithelial cells.

Animals↗

Kinetoplast DNA minicircles of phloem-restricted Phytomonas associated with wilt diseases of coconut and oil palms have a two-domain structure.

We report the cloning and sequencing of the first minicircle from a phloem-restricted, pathogenic Phytomonas sp. (Hart 1) isolated from a coconut palm with hartrot disease. The minicircle possessed a two-domain structure of two conserved regions, each containing three conserved sequence blocks (CSB). Based on the sequence around CSB 3 from Hart 1, PCR primers were designed to allow specific amplification of Phytomonas minicircles. This primer pair demonstrated specificity for at least six groups of plant trypanosomatids and did not amplify from insect trypanosomatids. The PCR results were consistent with a two-domain structure for other plant trypanosomatids.

Animals↗

As a bacterial culture medium, citrated sheep blood agar is a practical alternative to citrated human blood agar in laboratories of developing countries.

Human blood agar (HuBA) is widely used in developing countries for the isolation of bacteria from clinical specimens. This study compared citrated sheep blood agar (CSBA) and HuBA with defibrinated horse blood agar and defibrinated sheep blood agar (DSBA) for the isolation and antibiotic susceptibility testing of reference and clinical strains of Streptococcus pneumoniae, Streptococcus pyogenes, and Staphylococcus aureus. Reference and clinical strains of all organisms were diluted in brain heart infusion and a clinical specimen of cerebrospinal fluid and cultured on all agars. Viable counts, colony morphology, and colony size were recorded. Susceptibility testing for S. pneumoniae and S. pyogenes was performed on defibrinated sheep blood Mueller-Hinton agar, citrated sheep blood Mueller-Hinton agar (CSB MHA), and human blood Mueller-Hinton agar plates. For all organisms, the colony numbers were similar on all agars. Substantially smaller colony sizes and absent or minimal hemolysis were noted on HuBA for all organisms. Antibiotic susceptibility results for S. pneumoniae were similar for the two sheep blood agars; however, larger zone sizes were displayed on HuBA, and quality control for the reference strain failed on HuBA. For S. pyogenes, larger zone sizes were demonstrated on HuBA and CSBA than on DSBA. Poor hemolysis made interpretation of the zone sizes difficult on HuBA. CSBA is an acceptable alternative for the isolation of these organisms. The characteristic morphology is not evident, and hemolysis is poor on HuBA; and so HuBA is not recommended for use for the isolation or the susceptibility testing of any of these organisms. CSB MHA may be suitable for use for the susceptibility testing of S. pneumoniae.

Agar↗

The effect of glycosaminoglycans with acetaldehyde on the activation of prothrombin.

Heparin17-19k, (25, 50, and 100 ng), heparin6k (50 and 100 ng), heparin3k (50, 100, and 200 microg), chondroitin sulfates B (dermatan sulfate) (0.25, 0.5, and 1.0 microg), C (1 and 10 microg), and A (1 and 10 microg) each prolong the activated partial thromboplastin time (APTT) when preincubated with prothrombin to a greater extent than when preincubated with Factor II-deficient plasma prior to their mixing and subsequent additions of APTT reagent and Ca2+. In all cases statistical significance (p < or = 0.05) was observed except with the 2 lower levels of heparin3k. These results suggest that the glycosaminoglycans (GAGs) may exert a direct effect upon prothrombin (FII) in their anticoagulant activity. Pre mix tures of [(FII/25 ng H17-19k) + 447 mmol acetaldehyde (AcH)/L] as well as [(AcH/H) + FII] and [(FII/AcH) + H] each exert a synergistic anticoagulant effect upon APTT. At low AcH concentrations (44.7 mmol/L), neither a synergistic nor an additive effect is seen. H6k and H3k, on premixing with 447 mmol AcH/L, exhibit an additive effect on APTT prolongation but no synergism. Similarly, premixtures of CSB/447 mmol AcH/L/FII show a greater anticoagulant effect than do [(CSB/AcH) + FII] or [(FII/AcH) + CSB] premixtures. CSC-AcH and CSA-AcH patterns are analogous to those of CSB (DS). These data suggest the possibility that AcH, the primary product of ethanol metabolism, may serve as a crosslinking adduct with proteins, in this case, prothrombin, as well as GAGs. Thus ternary complexes between the zymogen form of coagulation factors, GAGs, and AcH are possible, further influencing coagulopathy.

Acetaldehyde↗

Spinobulbar neurons in lamprey: cellular properties and synaptic interactions.

An in vitro preparation of the nervous system of the lamprey, a lower vertebrate, was used to characterize the properties of spinal neurons with axons projecting to the brain stem [i.e., spinobulbar (SB) neurons)]. To identify SB neurons, extracellular electrodes on each side of the spinal cord near the obex recorded the axonal spikes of neurons impaled with sharp intracellular microelectrodes in the rostral spinal cord. The ascending spinal neurons (n = 144) included those with ipsilateral (iSB) (63/144), contralateral (cSB) (77/144), or bilateral (bSB) (4/144) axonal projections to the brain stem. Intracellular injection of biocytin revealed that the SB neurons had small- to medium-size somata and most had dendrites confined to the ipsilateral side of the cord, although about half of the cSB neurons also had contralateral dendrites. Most SB neurons had multiple axonal branches including descending axons. Electrophysiologically, the SB neurons were similar to other lamprey spinal neurons, firing spikes throughout long depolarizing pulses with some spike-frequency adaptation. Paired intracellular recordings between SB and reticulospinal (RS) neurons revealed that SB neurons made either excitatory or inhibitory synapses on RS neurons and the SB neurons received excitatory input from RS neurons. Mutual excitation and feedback inhibition between pairs of RS and SB neurons were observed. The SB neurons also received excitatory inputs from primary mechanosensory neurons (dorsal cells), and these same SB neurons were rhythmically active during fictive swimming, indicating that SB neurons convey both sensory and locomotor network information to the brain stem.

Animals↗

Effects of lesions of nucleus taeniae on appetitive and consummatory aspects of male sexual behavior in Japanese quail.

Neurochemical, hodological and functional criteria suggest that the nucleus taeniae and parts of the adjacent archistriatum represent the avian homologue of parts of the mammalian amygdaloid complex. It has been proposed in particular that the nucleus taeniae is the homologue of the mammalian medial amygdala. In male quail, relatively large lesions to the posterior/medial archistriatum selectively decrease the expression of appetitive sexual behavior in a manner reminiscent of similar manipulations involving the medial amygdala in mammals. We investigated the effects of discrete lesions restricted to nucleus taeniae and of lesions to an adjacent part of the archistriatum (pars intermedium ventralis, AIv) on the expression of appetitive (ASB) and consummatory (CSB) aspects of male sexual behavior. ASB was measured by a learned social proximity response (after copulation a male quail stands in front of a window providing visual access to a female) and by the frequency of rhythmic cloacal sphincter movements. CSB was assessed by the frequency of mount attempts (MA) and cloacal contact movements (CCM). Lesions confined to nucleus taeniae and to AIv did not influence the acquisition or the maintenance of the two responses indicative of ASB. In contrast, lesions of nucleus taeniae significantly increased the occurrence frequencies of MA and CCM when administered before the beginning of behavior testing and increased the frequency of MA only when performed on sexually experienced subjects. No effect of AIv lesions could be detected. The discrepancy between these results and previous experiments in quail might reflect procedural differences, but more probably differences in locations of the lesions that were restricted in the current study to the anterior part of taeniae. Those in the Thompson study were in the posterior part of this nucleus. These findings indicate that there is a larger degree of functional heterogeneity in the nucleus taeniae than previously thought. The effects of taeniae lesions suggest that this nucleus, similar to the medial amygdala in mammals, might be implicated in the control of sexual satiety.

Amygdala↗

Histamine release from human leukocytes: modulation by a cytochalasin B-sensitive barrier.

Cytochalasin B, a metabolic product of several fungi, enhances up to 10-fold the sensitivity and reactivity of human leukocytes to antigen E or anti-IgE-mediated histamine release. The effect of cytochalasin B is a result of its action on the second, antigen-independent, stage of histamine release. These data suggest that normally, antigen-triggered histamine release is modulated by a cytochalasin-sensitive barrier (CSB). This CSB modulation of histamine release can be separated from the modulating effect of cyclic adenosine monophosphate (AMP).

Antigens↗

Re-inventing the wheel; the use of autologous and fresh donor blood in cardiac surgery.

From the opening of a new cardiac surgical programme in November 1992, autologous and fresh donor blood (FDB) were used rather than cold stored blood (CSB) wherever possible in patients undergoing operations involving the use of cardiopulmonary bypass (CPB). In the first 250 consecutive patients, autologous blood was used in 168 (67.2%), fresh blood was used in 188 (75.2%). A total of 740 units of fresh blood were obtained on the day of operation (mean 3.9 +/- 1.6 units per patient able to supply donors; 4.9 +/- 1.7 units in the 147 who received fresh blood) and 728 units of stored blood were used (mean 3.08 +/- 1.84 units per patient where fresh blood was used; 6.2 +/- 2.5 units in the 114 where no fresh blood was used). The use of autologous blood significantly reduced FDB and CSB requirements (p < 0.001), was associated with a shorter intensive care and total postoperative stay (p = 0.006 and p = 0.033 respectively), even though there were more urgent and emergency cases in this group (p = 0.009) and no significant difference in chest drainage. Coagulopathy developed in 41 patients (16.4%) and was significantly associated with bypass time (p = 0.0001) and preoperative renal dysfunction (p = 0.005), although not with advanced age, sex, redo operation, diabetes or glucose-6-phosphate dehydrogenase deficiency. Patients with coagulopathy had significantly more transfused blood and blood products (p = 0.0001) and longer intensive care and total postoperative stays (p = 0.0001). In terms of blood conservation, the use of autologous blood was of primary importance.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Sequence variation and structural conservation in the D-loop region and flanking genes of mitochondrial DNA from Japanese pond frogs.

The nucleotide sequences of the D-loop region and its flanking genes of the mitochondrial DNA (mtDNA) from Japanese pond frogs were determined by the methods of PCR, cloning, and sequencing. The frogs belonged to two species, one subspecies, and one local race. The gene arrangements adjacent to the D-loop region were analyzed. The frogs shared a unique mitochondrial gene order that was found in Rana catesbeiana; i.e., cyt b--D-loop region--tRNA(Leu(CUN))--tRNA(Thr)--tRNA(Pro)--tRNA(Phe)--12S rRNA. The arrangements of the three tRNA genes of these frogs were different from those of X. laevis, a species which has the same overall structure as in mammals. Highly repetitive sequences with repeat units (16-bp or 17-bp sequence specific for each taxon) were found in the D-loop region. The length of repetitive sequences varied from 0.6 kbp to 1.2 kbp, and caused the extensive size variation in mtDNA. Several short sequence elements such as putative TAS, OH, CSB-1, and CSB-2 were found in the D-loop region of these frogs. The sequences of these short regulatory elements were conserved in R. catesbeiana, X. laevis, and also in human. The comparison of sequence divergences of the D-loop region and its adjacent genes among various taxa revealed that the rates of nucleotide substitutions depend on genes. The nucleotide sequences of the 3'-side segment of the D-loop region were the most variable among taxa, whereas those of the tRNA and 12S rRNA genes were the most conservative.

Animals↗

Production of bacterial cellulose from alternate feedstocks.

Production of bacterial cellulose by Acetobacter xylinum ATCC 10821 and 23770 in static cultures was tested from unamended food process effluents. Effluents included low-solids (LS) and high-solids (HS) potato effluents, cheese whey permeate (CW), or sugar beet raffinate (CSB). Strain 23770 produced 10% less cellulose from glucose than did strain 10821 and diverted more glucose to gluconate. Unamended HS, CW, and CSB were unsuitable for cellulose production by either strain, and LS was unsuitable for production by strain 10821. However, strain 23770 produced 17% more cellulose from LS than from glucose, indicating that unamended LS could serve as a feedstock for bacterial cellulose.

Acetobacter↗

Shear bond strength of different adhesive systems to primary dentin and enamel.

The aims of this study were to evaluate the shear bond strength (SBS) off our adhesive systenms applied to primary dentin and enamel and verify, after SBS testing, the failure mode of the adhesive interface. Sixty extracted sound primary molars were selected and crowns were sectioned in a mesial-distal direction. Specimens were randomly assigned into two groups (adhesion to enamel and adhesion to dentin) and then subdivided into four subgroups according to the adhesive system (n=15). Scotchbond Multi-Purpose (SMP)--Single Bond (SB)--Clearfil SE Bond (and Adper Prompt L-Pop (APL)--SBS tests were performed and the obtained values were statistically analyzed using ANOVA and Tukey tests (p < 0.05). The failure mode analysis was performed with a Scanning Electron Microscope (XL-30, Philips). SBS mean values on enamel were [MPa (SD)]: SMP--2789 (749); SB--23.92 (8.8); CSB--24.36 (6.69), APL--25.96 (4.08); and on dentin: SMP--17.29 (4.25) SB--18.2 (8.74); CSB--16.13 (714); APL--6. 04 (3.35). The predominant failure mode was cohesive (primarily of the bonding agent). On enamel SBS was statistically similar for all four adhesives. On dentin SBS ofAPL was lower than the other tested adhesives.

Bisphenol A-Glycidyl Methacrylate↗

Effect of diet and population density on male turkeys under various environmental conditions. 1. Turkey growth and health performance.

The performance of 1,312 male market turkeys (Large White, Nicholas strain) from 0 to 20 wk of age fed diets varying in feed form and energy level was measured under two stocking densities (.21 or .46 m2 per bird) and four lighting and temperature programs. The four diets were 1) corn and soybean meal with 1% supplemental fat, mash (CSM); 2) as 1, pelleted (CSP); 3) as Diet 1 but with 1, 2, 4, 6, and 8% supplemental fat during 0 to 4, 4 to 8, 8 to 12, 12 to 16, and 16 to 20 wk of age, respectively (CSF); and 4) as Diet 1 but with barely included at 0, 20, 35, 50, and 65% during the respective 4-wk age periods (CSB). The four light and temperature programs were 1) Environment A with intermittent light [4 [2 h light (L):4 h dark (D)]] in combination with cycling temperature at 7 and 21 C during light and dark photoperiod, respectively; Environment B with intermittent light, 21 C; Environment C with continuous light (18L:6D) and cycling temperature of 7 to 21 C; and Environment D with intermittent light, 7 C. Lighting and temperature programs started at 1 and 4 wk of age, respectively. Body weights at 20 wk of age decreased (P less than .05) with increasing temperature (13.86 versus 12.26 kg for Environments D and B, respectively) with cycling temperature intermediate (13.51 kg for Environment A). Intermittent light (P less than .05) improved BW and feed conversion by 3.4 and 2.0%, respectively, compared with continuous light. Rearing males at .21 m2 per bird versus .46 m2 per bird decreased weight (P less than .05) by 5.5%. Twenty-week BW of males fed the CSP (13.52 kg) and CSF (13.58 kg) diets were greater (P less than .05) than those fed CSM (12.90 kg) and CSB (12.69 kg) diets. Significant (P less than .05) interactions between diet, environment, and density were not detected for most performance characteristics. Environmental measurements indicated higher dust and ammonia levels in the warm environment (B). Isolates of aspergillus and incidence of airsacculitis at time of processing were greatest in Environment B.

Air Sacs↗

Evidence for in the appearance of mRNAs of nucleotide excision repair genes, in human ovarian cancer tissues.

Nucleotide excision repair (NER) is the DNA repair pathway through which cisplatin-DNA intrastrand adduct is repaired. Clinical studies have shown that increased mRNA expression of selected genes involved in the rate-limiting step of NER, appear to be closely associated with clinical resistance to platinum agents. These specific studies have led to the possibility of an assessment of the order in which, the appearance of mRNAs of selected NER genes may occur. Included in this assessment are ERCC1, XPB, CSB, and XPA, studied in 28 ovarian cancer tumor tissue specimens. The study of these four genes, in pairs, from 28 ovarian cancer specimens, results in 168 separate observations. Based on the mRNA expression patterns of these genes in these tissues, it is suggested that ERCC1 mRNA may appear before the mRNA of any of the other genes, in an obligate fashion. This is followed by XPB mRNA; which appears before the mRNA of XPA; which in turn, may appear before CSB. This pattern is consistent with what we have reported previously, in non-malignant human bone marrow specimens from a cohort of 52 patients.

Adenocarcinoma↗

Control of epidemic malaria on the highlands of Madagascar.

The Malagashy national malaria control programme ('Programme National de Lutte contre le Paludisme', PNLP) has been developing, since 1996, an epidemiological early warning system for malaria epidemics in the Central Highlands with the support of the Italian Development Cooperation. The system is based on the monitoring of malaria morbidity (clinical diagnosis) in 536 peripheral health centres (CSB) of the Highlands. The intervention area corresponds to 27 districts of the Antananarivo and Fianarantsoa provinces (4.7 million inhabitants) and spans around 100,000 square km. For each CSB a monthly warning threshold, defined as the 1993-1996 monthly mean number of malaria cases plus two standard deviations, was established. Three levels of epidemic alert have been defined according to the number of times the cases of presumptive malaria surpassed the threshold and according to the reported presence of severe malaria cases. The surveillance system relies also on the monitoring, in district hospitals of the Highlands, of the Plasmodium falciparum infection rate among clinically diagnosed malaria cases. A total of 185,589 presumptive malaria cases, corresponding to a 42/1000 malaria incidence, were recorded in 1997 by the surveillance system. During the same year 184 alerts of 2nd degree were reported. During 1998 173,632 presumptive malaria cases corresponding to a 38/1000 incidence were reported and 207 alerts of 2nd degree were detected; 75 of these alerts were investigated with ad hoc surveys and 3 initial malaria epidemics identified and controlled. Out of 6884 presumptive malaria cases diagnosed in the district hospitals during 1997-1998, only 835 (12.1%) have been confirmed by microscopy (P. falciparum 81.7%, P. vivax 15.0%, P. malariae 2.5%, P. ovale 0.2%, mixed infections 0.6%); 22.4% of these infections were imported cases from coastal endemic areas. The efficiency of the system in monitoring the trend of malaria morbidity and in the rapid detection and response to malaria epidemics is still being evaluated.

Animals↗

[Micromethod for identification of enterobacteria].

The aim of this study was to set accurate and reliable methods in the identification of Enterobacteriaceae. In Micro CSB Entero each of a strip with 20 cupules containing dehydrated substrates for biochemical identification of bacterial species. Strips were inoculated with 100 microliters per cupule and incubated. After 18 hours, reagents were added. Baye's theorem was used to validate tests. Reactions from Micro CSB were cleared and easily read. 102 strains of Enterobacteriaceae were identified by MicroCSB--89.3% were correctly identified. 0.9% were identified with other tests 9.8% were incorrectly identified MicroCSB gives many advantages: This method save time, space and is less expensive than the other methods and it gives security for identifying species belonging to the different genera.

Bacterial Typing Techniques↗

Differential binding affinity of immobilized Concanavalin A-Sepharose 4B for normal and myelomatous immunoglobulins.

Immobilized Concanavalin A-Sepharose 4B (CSB) binds myeloma immunoglobulins (IgG through IgE) in larger proportions than it binds normal immunoglobulins. Some possible explanations for this are discussed. Also, CSB shows differential affinity among immunoglobulins, normal or myelomatous; its greatest affinity is for IgM and IgE. This property can be used to separate a large percentage of the IgM and IgE from both normal and myelomatous sera.

Humans↗

Application of low-cost extrusion cooking to weaning foods in feeding programmes. part 2.

A low-cost technology for making blended foods and oilseed flours has been developed within a programme operated by Colorado State University, USA. The technology is based on a low-cost extruder cooker (LEC). The primary products are 1) vitamin- and mineral-fortified cereal/legume blends for use in supplementary feeding programmes, and 2) protein supplements for fortifying cereal and/or root crop foods. Plants have been installed in four developing countries, and research and development activities are being carried out in a number of others. The programme involves agricultural and industrial development as well as nutritional improvement. The major product currently made is CSB, and in some cases CSM made by blending dried milk with CSB. These products have been demonstrated to be safe, nutritious and acceptable by the target population, although more work is needed on expanding the ways in which the blended foods might be used in traditional recipes, as well as in gruels and porridges.

Child, Preschool↗

Cyclosporin H is a potent and selective formyl peptide receptor antagonist. Comparison with N-t-butoxycarbonyl-L-phenylalanyl-L-leucyl-L-phenylalanyl-L- leucyl-L-phenylalanine and cyclosporins A, B, C, D, and E.

The cyclic undecapeptide, cyclosporin (Cs) H, is a potent inhibitor of FMLP-induced superoxide anion (O2-) formation in human neutrophils. We studied the effects of CsH in comparison with those of N-t-butoxycarbonyl-L-phenylalanyl-L-leucyl-L-phenylalanyl-L-leucyl-L- phenylalanine (BocPLPLP), a well known formyl peptide receptor antagonist, and of other Cs on activation of N6,2'-O-dibutyryl adenosine 3:5'-monophosphate-differentiated HL-60 cells and human erythroleukemia cells (HEL cells). CsH inhibited FMLP binding in HL-60 membranes with a Ki (inhibition constant) of 0.10 microM. CsH inhibited activation by FMLP of high affinity GTPase (the enzymatic activity of alpha-subunits of heterotrimeric regulatory guanine nucleotide-binding proteins) in HL-60 membranes with a Ki of 0.79 microM. CsH inhibited the stimulatory effects of FMLP on cytosolic Ca2+ concentration ([Ca2+]i), O2- formation, and beta-glucuronidase release with Ki values of 0.08, 0.24, and 0.45 microM, respectively. BocPLPLP was 14-fold less potent than CsH in inhibiting FMLP binding and 4- to 6-fold less potent than CsH in inhibiting FMLP-induced GTP hydrolysis, rises in [Ca2+]i, O2- formation, and beta-glucuronidase release. CsA reduced FMLP-induced O2- formation by 20%, but CsB, CsC, CsD, and CsE did not. CsA, CsB, CsC, CsD, and CsE did not affect FMLP-induced rises in [Ca2+]i. BocPLPLP inhibited leukotriene B4-induced rises in [Ca2+]i with a Ki of 0.33 microM, whereas CsH showed no inhibitory effect. CsH and BocPLPLP did not inhibit the rises in [Ca2+]i induced by several other stimuli in HL-60 cells and HEL cells. Our results show that 1) CsH is a more potent formyl peptide receptor antagonist than BocPLPLP; 2) unlike BocPLPLP, CsH is selective; and 3) N-methyl-D-valine which is present at position 11 of the amino acid sequence of CsH but not of other Cs is crucial for FMLP antagonism.

Amino Acid Sequence↗