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PubMed2002

Complete genomes of two Human hepatitis A virus isolates from China: analysis and comparison with other isolates.

Complete sequences of the genomes of two wild type (wt) Human hepatitis A virus (HHAV) isolates, LU38 and LY6 from China were determined and compared with those of wt HHAV isolates AH1, AH2, AH3, FH1, FH2, FH3, GBM, HM175, LA and MBB. The genomes of both LU38 and LY6 consisting of 7477 nucleotides (nts) contained a 5'-non-translated region (5'-NTR, 733 nts), an open reading frame (ORF, 6681 nts), and a 3'-NTR (63 nts) followed by a poly(A)-tail. It encoded a polyprotein of 2227 amino acids (aa) Sequence comparison showed that LU38 shared the highest identities of 98.1% for nt (140 differences) and 99.2% for as (17 differences) with AH1, and the lowest identities of 91.4% for nt (741 differences) with HM175 and 98.1% for aa (43 differences) with GBM. LY6 shared the highest identities of 97.4% for nt (196 differences) and 98.7% for aa (28 differences) with H1 and the lowest identities of 91.2% for nt (642 differences) with HM175 and 97.7% for as (51 differences) with GBM. The subgenotyping revealed that the LU38 and LY6 isolates are of IA subgenotype. The phylogenetic analysis showed that LU38 is closest to AH1 and the LY6 to FH3, suggesting that the epidemiological link of hepatitis A (HA) had developed in China and Japan.

Amino Acid Sequence↗
PubMed2004

[The isolation frequency and antimicrobial susceptibility of Haemophilus influenzae isolated in Saga University Hospital].

Isolation frequency and antimicrobial susceptibility of Haemophilus influenzae isolated in Saga University hospital from October 2002 to September 2003 were investigated. Out of 155 H. influenzae strains subjected 77 were isolated from pediatrics specimens. beta-Lactamase negative ampicillin (ABPC)-resistant H. influenzae (BLNAR), against which MICs of ABPC were higher than 4 microg/mL, were 32 strains (20.6%), and it became 63 strains (41.3%) when Low-BLNAR, against which MICs of ABPC were higher than 2 microg/mL, were included. beta-Lactamase positive ABPC-resistant H. influenzae (BLPAR) were 8 strains (5.2%). Although those BLNAR were also resistant to variety of beta-lactams, fluoroquinolones and other antibiotics were not affected by the level of ABPC-resistance. Resistant strains of BLPAR against SBT/ABPC, a combination of a beta-lactamase inhibitor, were detected but all of them were sensitive to TAZ/PIPC, an another combination. Those strains were able to be considered as beta-lactamase positive amoxicillin-clavulanate resistant H. influenzae (BLPACR). PIPC, TAZ/PIPC, CTRX, CDTR, MEPM, LVFX and CPFX showed good activity among tested antibiotics.

Adolescent↗
PubMed2004

Human leptospirosis in Erode, South India: serology, isolation, and characterization of the isolates by randomly amplified polymorphic DNA (RAPD) fingerprinting.

The study describes the first attempt to record leptospirosis in Erode by isolation and serological tests such as the microscopic agglutination test and IgM-based enzyme-linked immunosorbent assay. Twenty-nine clinically suspected cases showing fever, headache, body ache associated with jaundice, decreased urine output, and conjunctival suffusion were included. The age of the patients ranged between 10-71 years and most of them were agricultural workers. Paired sera were possible among 12 cases. All the patients had fever and headache and other more common symptoms were myalgia and icterus. Leptospiral culture was positive in 7 (24.1%) patients. Out of 29 patients, 26 (89.7%) were diagnosed as having current leptospiral infection based on serology and isolation. The leptospiral isolates KSR 1-6 were further characterized by using randomly amplified polymorphic DNA fingerprinting shown genetic similarities with Leptospira interrogans spp. This study shows the presence of leptospirosis among the hospital cases of Erode and that this disease is a potential health hazard of agricultural workers in Cauvery basin.

Adolescent↗
PubMedDate not supplied

[Isolation and characterization of bovine diarrhea viral isolates].

The paper presents the results of a study of the antigenic and electromicroscopic characteristics of 3 bovine viral diarrhea isolates from cattle in Siberia. All the isolates were antigenically related to the reference strain BK-1 and closely interrelated: their affinity was in the range of 92.2 to 96.4%. The aerosolic administration of the isolate of TM from the sick calf lung into 2 seronegative (4-6-month-old) calves caused the characteristic sings of acute respiratory disease with short diarrhea.

Animals↗
PubMed1990

Effect of procaterol on the isolated airway smooth muscle and the release of anaphylactic chemical mediators from the isolated lung fragments.

The effects of an orally active and selective beta 2-stimulant, procaterol (OPC-2009) on the isolated pulmonary smooth muscle and the release of chemical mediators from the passively sensitized lung fragments were studied and compared with those of isoprenaline (isoproterenol) and salbutamol. Procaterol potently relaxed the isolated guinea pig trachea and lung parenchyma in a concentration-dependent fashion. The drug at a similar range of concentrations antagonized the contraction of the isolated guinea pig trachea and lung parenchyma or human bronchus induced by leukotriene (LT) D4. Salbutamol and isoprenaline also showed similar effects to those of procaterol on the above experiments, but the potencies were consistently weaker than that of procaterol. The release of either histamine or LTs from the passively sensitized human lung fragments was markedly and dose-dependently inhibited by both the 5 min and 15 h treatment with procaterol (10(-10)-10(-7) mol/l) before antigen challenge. Isoprenaline and salbutamol in 5 min treatment experiments showed similar potency as and less potency than procaterol, respectively, on the release of these mediators, but the inhibition potencies of these drugs, particularly isoprenaline, were remarkably reduced by 15 h treatment. From these results, procaterol, besides the existing use as a bronchodilator, is expected to be a potentially prophylactic drug for allergic asthma because of the strong inhibition of the anaphylactic mediator release.

Albuterol↗
PubMed2006

Isolation and characterization of nickel uptake by nickel resistant bacterial isolate (NiRBI).

OBJECTIVE: Bioremediation technology has gained importance because microbes could be the convenient source of bio-absorption/bioaccumulation of metals from effluent streams. METHODS: The nickel-resistant bacterial isolates (NiRBI) were selected from various bacterial isolates from industrial effluent and grown in nutrient broth containing different concentrations of nickel sulfate (0.3-3.0 mmol/L) and their capability of accumulating metal from the medium. RESULTS: Well-defined growth of NiRBI was observed in the medium containing up to 2.5 mmol/L of nickel. The isolate was identified using 16S rRNA and closely related to Pseudomonas fragi. Maximum accumulation of nickel (0.59 mg/g dry weight of bacterial cells) was observed when NiRBI was grown in media containing 2 mmol/L of nickel. The protein profile of the NiRBI cellular extract by SDS-PAGE showed two metal stress-induced proteins of molecular weight 48 KD and 18 KD with a simultaneous down regulation of four proteins of 46.7 KD, 42.2 KD, 19.7 KD, and 4.0 KD. CONCLUSION: 48 KD and 18 KD proteins play a role in metal resistance mechanism by NiRBI.

Biodegradation, Environmental↗
PubMedDate not supplied

Identification of seven isolants of infectious bronchitis virus by interference with the B-1 isolant of Newcastle disease virus.

Seven isolants of infectious bronchitis virus (IBV) interfered with the development of hemagglutinins (HA) by the B-1 isolant of Newcaslte disease virus (NDV) in eggs previously inoculated with these IBV isolants. The I50 [defined as the minimum amount of IBV needed to produce, upon inoculation in embryonating chicken eggs (ECE), HA interference in 50% of these ECE reioculated 10 hr later with 10(2,41 EID50 (HA) of the B-1 strain of NDV] was less than 1 EID50. The interference was considered specific for IBV because if could be eliminated by addition of homologous anti-IBV serum. Furthermore, three other common poultry viruses (laryngotracheitis, fowlpox, and CELO viruses) did not interfere with NDV. In view of its sensitivity, specificity, and simplicity of performance, the IBV-NDV interference test appears to have considerable potential as a diagnostic test for IBV under field conditions.

Coronaviridae↗
PubMed1990

Nonlinear elimination kinetics of 5-fluoro-2'-deoxyuridine in isolated perfused rat liver and isolated hepatocytes.

The kinetic parameters of the cytostatic agent 5-fluoro-2'-deoxyluridine (FUDR) were studied in isolated rat hepatocytes and in the isolated perfused rat liver. In both experimental setups a dose dependency of the elimination parameters, half-life and clearance, was observed with a calculated turning point around 250 microM. In the medium of rat hepatocytes incubated at low (0.1 microM) to high (2000 microM) FUDR, the majority of the metabolites consisted of the catabolite alpha-fluoro-beta-alanine. The nucleobase metabolites, 5-fluorouracil and its primary product 5,6-dihydro-5-fluorouracil, approached apparent steady-state levels comprising 10 to 15% of the initial concentration. In the intracellular phase of hepatocytes incubated at 300 microM FUDR almost 90% of the FUDR-derived material was alpha-fluoro-beta-alanine, whereas essentially no unchanged FUDR could be detected. Similar results were obtained at extracellular FUDR concentrations exceeding 300 microM. In the isolated perfused rat liver, the clearance decreased to 15 to 20% of the corresponding values when the initial concentration was raised from 24 to 2400 microM. At the end of perfusion alpha-fluoro-beta-alanine comprised 90 to 95% of FUDR-derived total radioactivity in the tissue even at initially 2400 microM FUDR, although at this FUDR dosage 20% of the substrate remained unmetabolized in the medium. These results suggest that the limitation of hepatic FUDR elimination is not due to saturable hepatic metabolism but must be due to saturable uptake of these pyrimidine derivatives across the cellular membrane of parenchymal liver cells.

Animals↗
PubMed1985

A regional analysis of alpha-spectrin in the isolated Mauthner neuron and in isolated axons of the goldfish and rabbit.

Isolated dendrites, somata, and desheathed axons of the goldfish Mauthner neuron (M-cell), in addition to other isolated myelin sheath-free axons of the goldfish spinal cord and of rabbit lumbar ventral roots, were shown by immunochemical and immunofluorescence techniques to contain alpha-spectrin (fodrin). alpha-Spectrin appeared to be organized as a randomly distributed reticular network, localized to the surface of isolated neuronal cellular structures. In addition, alpha-spectrin was also distributed nonrandomly at specialized cellular sites. These sites included synaptic junctions and morphologically differentiated nodes of Ranvier (i.e., rabbit axons, but not goldfish axons). At the latter sites, it is possible to demonstrate that alpha-spectrin is co-localized with F-actin, as indicated by a striking correspondence of fluorescent images due to double labeling, using the indirect immunofluorescence technique with alpha-spectrin antiserum, and direct binding of F-actin by rhodamine-conjugated palloidin. However, the spectrin-actin network at synaptic junctions appears to be distributed over the entire area of junctional contact and is not just restricted to postsynaptic densities. The possibility of a duality of roles of spectrin in membrane-related motile and anchorage functions is discussed.

Animals↗
PubMed1988

Further isolation of HIV strains in Palermo: some biological properties of the new isolates.

21 HIV strains were isolated from peripheral mononuclear cells of 31 adult and pediatric patients with AIDS or ARC and 10 apparently healthy HIV seropositive individuals. Viral isolation rate was correlated to the clinical status of patients, being very high in patients with AIDS or ARC, and lower in apparently healthy individuals. 4 isolates were propagated in cultures of a T-cell continuous line (Molt 4) and their biological properties were compared. The capacity to establish a chronic infection on Molt 4 cells and the extent of cytopathic effect of the infected cultures varied among the different strains. Moreover, differences in endpoint titrations of the supernatants and variable effects on cell multiplication and viability of Molt 4 cells were also observed. Our results further substantiate the existence of a wide heterogeneity of HIV strains.

AIDS-Related Complex↗
PubMed1986

[Frequency of isolation and drug resistance of bacterial strains isolated from respiratory material in patients with cystic fibrosis].

During 1985, 251 respiratory samples from 61 patients were examined at the tuscan cystic fibrosis Center (Florence), and isolated strains were tested against various antimicrobial drugs. Pseudomonas aeruginosa and Staphylococcus aureus were the predominant pathogens isolated. Infections caused by St. aureus were the commonest during the first years of life, whereas those caused by Ps. aeruginosa, occasional at early age, became chronic and the main source of infection with growth. Sensitivity testings against antibiotics confirmed that Ps. aeruginosa isolated from cystic fibrosis patients has a high percentage of resistant strains. The most active drugs were ceftazidime and aztreonam. An increased resistance to aminoglycosides was observed.

Adolescent↗
PubMedDate not supplied

Studies on orthoreoviruses isolated from young turkeys. I. Isolation and characterization.

Four virus isolates were recovered from the intestinal tracts of turkey poults with diarrhea, abnormal feathering, stunted growth, and increased mortality. The isolates were clone-purified and characterized according to cytopathic effect (CPE) in chicken embryo kidney cells, size, morphology, physicochemical properties, and replicative cycle. The isolates were identified as members of the family Reoviridae, genus Orthoreovirus.

Animals↗
PubMed1980

Structure of a newly isolated variant of Simian virus 40 DNA containing monkey DNA and its similarity to previously isolated variants.

The structure of a newly and independently isolated defective variant of simian virus 40 that contains covalently linked monkey and SV40 DNA sequences is described. This variant, termed 290, has a structure essentially identical with a previously isolated and characterized variant named CVP8/1/P2 (Eco RI res). The structural similarities include the monkey (host) DNA segment that is combined with viral DNA sequences, the particular viral DNA segment that is present, and the arrangement of these within the defective genome. The monkey DNA segment contains sequences derived from both low and high reiteration frequency monkey DNA. The viral sequences include the origin of replication. The separate isolation of essentially identical variants suggests a high level of specificity in the events leading to the formation and amplification of this type of defective genome.

Animals↗
PubMed1981

Isolation and significance of anaerobic bacteria isolated from cases of bovine mastitis.

The role of obligate anaerobic bacteria in the aetiology of mastitis of lactating dairy cows was investigated. Anaerobes were isolated from 12% of lactating mastitic cows, which were representative of 50% of the 10 dairy herds examined. Bacteroides fragilis was the most frequently isolated organism of (50%), followed by Peptococcus indolicus (33%), Eubacterium lentum (33%), E. aerofaciens (17%), Propionibacterium granulosum (17%) and an anaerobic Streptococcus sp. (17%). These obligate anaerobes were always isolated together with organisms classically involved in mastitis. It was possible to induce overt clinical mastitis in healthy lactating udders within 24 hours by infection with single pure cultures of anaerobes via the teat canal. All B. fragilis strains were resistant to penicillin G and tetracycline. In addition, one strain was also resistant to ampicillin, cephalothin and amoxicillin. Anaerobic gram positive cocci and bacilli were sensitive to most antibiotics. These findings imply an important role for anaerobes in the aetiology of mastitis.

Animals↗
PubMed1995

Effect of Brefeldin A on transcytotic vesicular pathway and bile secretion: a study on the isolated perfused rat liver and isolated rat hepatocyte couplets.

This study investigated the effect of Brefeldin A (BFA) on the transcytotic vesicular pathway labeled with horseradish peroxidase (HRP) in both isolated rat hepatocyte couplets (IRHC) and the isolated perfused rat liver (IPRL). To evaluate the role of the transcytotic vesicular pathway on bile secretion, the effect of BFA on bile secretion in the IPRL was then investigated. In the basolateral area of IRHC, BFA showed no effect on the density and percentage of area of HRP-labeled vesicles. However, HRP-labeled vesicles tended to accumulate in the juxtanuclear area of BFA-treated hepatocytes (P < .001 vs. controls). In the pericanalicular area, on the other hand, HRP-labeled vesicles were depleted compared with controls (P < .001). In keeping with these findings, although the early peak remained unchanged, BFA inhibited as much as 50% of the late peak of HRP excretion in bile, after a pulse load of HRP in the IPRL. Bile flow and the biliary secretion of bile salts (BS) and phospholipids were not modified by BFA in isolated livers perfused without BS in the perfusate or with 1 mumol/min taurocholate (TCA). In BFA-treated livers, peak bile flow and BS output decreased by 20% (P < .05 vs. controls) only when a 5 mumol TCA bolus was administered. In conclusion, this study demonstrates that BFA inhibits the transcytotic vesicular pathway in the liver. However, BFA has no significant effect on bile secretion either in basal conditions or during perfusion with physiological amounts of BS. BFA slightly decreases bile flow and BS output only after an overload of BS, providing evidence against the physiological relevance of the transcytotic vesicular pathway in the process of bile formation.

Animals↗
PubMedDate not supplied

Potassium channel-opening and vasorelaxant profiles of a novel compound YM099 in rat isolated portal vein and rabbit isolated aorta.

The potassium channel-opening and vasorelaxant profiles of YM099, a newly synthesized benzoxadiazol derivative K channel opener, were evaluated in vitro. In the rat isolated portal vein, YM099 and a benzopyran derivative K channel opener levcromakalim concentration-dependently inhibited the frequency of spontaneous rhythmic contractions, with IC50 values of 104 and 38 nM, respectively. In the rabbit isolated aorta, YM099 (10(-7)-3 x 10(-5) M) and levcromakalim (10(-7)-3 x 10(-5) M) also concentration-dependently relaxed the contractions induced by 20 mM KCl, but they were ineffective against the contractions induced by 50 mM KCl. These effects of YM099 and levcromakalim were competitively antagonized by a K channel blocker glibenclamide (10(-7)-3 x 10(-6) M). In the rabbit isolated aorta, YM099 (3 x 10(-8)-3 x 10(-6) M), but not the calcium antagonist nifedipine (10(-8)-3 x 10(-6) M), relaxed the contractions induced by norepinephrine (10(-6) M) or prostaglandin F2 alpha (3 x 10(-6) M). These vasorelaxant effects of YM099 were also antagonized by glibenclamide. In conclusion, YM099 is a potent vascular smooth muscle-relaxant agent and possesses a vasorelaxant effect different from that of nifedipine. These effects of YM099 may be mediated, like those of levcromakalim, by the opening of glibenclamide-sensitive K channels.

Animals↗
PubMed1996

[Antimicrobial activities of meropenem against clinically isolated strains. The result against strains isolated from blood and cerebrospinal fluid].

In order to evaluate the antimicrobial activity of meropenem (MEPM), minimum inhibitory concentrations (MICs) of MEPM and control drugs were determined against clinical isolates from blood and cerebrospinal fluid that were obtained from January, 1993 to December, 1994. The results are summarized as follows; 1. The MIC-range, 50% MIC (MIC50) and 90% MIC (MIC90) of MEPM were equal to those of imipenem (IPM) and panipenem (PAPM) against Streptococcus pneumoniae including benzylpenicillin (PCG)-insensitive or -resistant S. pneumoniae, Streptococcus agalactiae and Listeria monocytogenes which are Gram-positive strains, and were stronger than those of ampicillin (ABPC) and cefotaxime (CTX). 2. The MIC-range, MIC50 and MIC90 of these 3 drugs of carbapenems (MEPM, IPM and PAPM) were different against Escherichia coli and Haemophilus influenzae which are Gram-negative strains. The MIC90 of MEPM was < or = 0.025 microgram/ml and those of IPM and PAPM were 0.2 microgram/ml against E. coli. The MIC90 of MEPM was 0.1 microgram/ml, that of IPM was 25 micrograms/ml and that of PAPM was 6.25 micrograms/ml against H. influenzae. Thus, the antimicrobial activity of MEPM was stronger than those of IPM and PAPM. The MIC90's of IPM and PAPM against H. influenzae were high with the MIC of IPM at 12.5 approximately 25 micrograms/ml and the MIC of PAPM at 3.13 approximately 12.5 micrograms/ml against 3 IPM-resistant strains among 17 isolates. 3. The MIC90 of ABPC was 0.39 microgram/ml and that of CTX was 0.1 microgram/ml against 20 strains of S. pneumoniae including 6 strains of PCG-insensitive or resistant S. pneumoniae. The MIC90 of ABPC and CTX were higher than those of 3 carbapenem drugs. There were E. coli of 8 strains with ABPC-high resistance (the MIC of ABPC was > 100 micrograms/ml) and 2 strains for which MIC of CTX were 0.39 microgram/ml and 3.13 micrograms/ml. It was found that 29.4% of H. influenzae were beta-lactamase producing strains. 4. It appeared that antimicrobial activities of carbapenems, particularly MEPM were strong against clinical isolates from blood and cerebrospinal fluid. MEPM will be first choice drug by empiric therapy in infections including sepsis and purulent meningitis.

Ampicillin↗
PubMed1997

[Efficacy of combined use of sodium thiosulfate in percutaneous isolated liver chemoperfusion of cisplatin under complete hepatic venous isolated and charcoal hemoperfusion].

We herein evaluated the efficacy of combined use of sodium thiosulfate (STS) in isolated liver chemoperfusion of cisplatin (CDDP) under complete hepatic venous isolation and charcoal hemoperfusion (HVI.CHP). Dogs were divided into two groups; group I (n = 5), CDDP (4 mg/kg) + STS (4 g/body) and group II (n = 5), CDDP (4 mg/kg). In both groups, CDDP was intraarterially administered for 20 minutes under HVI.CHP. In addition, in group I, STS infusion was administered for 20 minutes via the outlet side of CHP filter. The adsorption rates of free and total CDDP of CHP filter were in a range of 50-80%, and the rates in group I were higher than in group II. At all sampling points, the AUC (microgram.min/ml) of free CDDP was not different between the two groups. However, the AUC of total CDDP at postfilter [group I: 35.3 +/- 2.7 (mean +/- SD); group II: 63.5 +/- 21.5] and systemic serum (group I: 24.9 +/- 8.6, group II: 43.5 +/- 13.5) was significantly lower in group I than in group II (p < 0.05). The cumulative amounts of urinary excretion of CDDP (microgram/kg) were 39.5 +/- 12.1 and 13.5 +/- 9.3 in groups I and II, respectively, showing a significant increase in group I (p < 0.01). These results indicated that the use of STS in percutaneous isolated liver chemoperfusion of CDDP under HVI.CHP could further reduce systemic toxicities of CDDP, by increasing CDDP filter extraction of CHP filter and urinary CDDP excretion.

Animals↗