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Biomedical subjects

A Ali

Publications and source records attributed to A Ali.

At least 235 records · Page 13Linked to original sources

Malonyl-coenzyme A:acyl carrier protein acyltransferase of Streptomyces glaucescens: a possible link between fatty acid and polyketide biosynthesis.

Streptomyces glaucescens, a Gram-positive soil bacterium, produces the polyketide antibiotic tetracenomycin (Tcm) C. To study possible biochemical connections between the biosynthesis of bacterial fatty acids and polyketides, the abundant acyl carrier protein (ACP) detected throughout the growth of the tetracenomycin (Tcm) C-producing S. glaucescens was purified to homogeneity and found to behave like many other ACPs from bacteria and plants (apparent M(r) of 20,000 on gel filtration chromatography, apparent M(r) of 3400-4800 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions, and pI approximately 3.8). By using an oligodeoxynucleotide synthesized in accordance with the sequence of residues 25-36 of the ACP, the fabC gene encoding this protein was cloned, and expression of this gene in Escherichia coli yielded the ACP entirely as the active holoenzyme. Sequence analysis of 4.3 kilobases (kb) of DNA flanking fabC revealed the presence of three other genes oriented in the same transcriptional direction in the order fabD, fabH, fabC, and fabB. Each of the four genes is predicted to encode proteins with high sequence similarity to the following components of the E. coli fatty acid synthase (FAS): the FabD malonyl-coenzyme A:ACP acyltransferase (MAT), FabH 3-oxoacyl:ACP synthase III, AcpP ACP, and FabB 3-oxoacyl:ACP synthase I. Expression of the S. glaucescens fabD gene in E. coli produced active MAT able to catalyze in vitro the transfer of radioactive malonate from malonyl-coenzyme A to the E. coli AcpP and S. glaucescens FabC ACPs, as well as to the TcmM ACP component of the Tcm type II polyketide synthase [Shen, B., et al. (1992) J. Bacteriol 174, 3818-3821]. Expression of fabD also restored the high-temperature growth of the E. coli fabD89 mutant that bears a temperature-sensitive MAT. The latter finding and the close similarity between the organization of the S. glaucescens fabDHCB and E. coli FAS-encoding genes (fabH/fabD/fabG/acpP/fabF) suggest that the S. glaucescens genes encode FAS enzymes. Moreover, on the basis of its in vitro activity, it is possible that the S. glaucescens FabD MAT is responsible for charging the TcmM ACP with malonate in vivo, a key step in the synthesis of the deca(polyketide) precursor of Tcm C. This implies the existence of a functional connection between fatty acid and polyketide metabolism in this bacterium.

Acyl-Carrier Protein S-Malonyltransferase↗

Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis.

The protease responsible for the cleavage of poly(ADP-ribose) polymerase and necessary for apoptosis has been purified and characterized. This enzyme, named apopain, is composed of two subunits of relative molecular mass (M(r)) 17K and 12K that are derived from a common proenzyme identified as CPP32. This proenzyme is related to interleukin-1 beta-converting enzyme (ICE) and CED-3, the product of a gene required for programmed cell death in Caenorhabditis elegans. A potent peptide aldehyde inhibitor has been developed and shown to prevent apoptotic events in vitro, suggesting that apopain/CPP32 is important for the initiation of apoptotic cell death.

Amino Acid Sequence↗

Molecular cloning and pro-apoptotic activity of ICErelII and ICErelIII, members of the ICE/CED-3 family of cysteine proteases.

Cysteine proteases related to mammalian interleukin-1 beta-converting enzyme (ICE) and the nematode cell death abnormal ced-3 gene product have been implicated in the effector mechanism of apoptotic cell death. Two novel members of this new family of ICE/CED-3-related proteases, designated ICErel-II and ICErel-III, were cloned from human monocytic cells. Both were highly homologous to human ICE (52% identical) and CED-3 (25% identical) and both contained the absolutely conserved pentapeptide sequence Gln-Ala-Cys-Arg-Asp containing the catalytic cysteine residue. Other structural motifs that were comparable with ICE suggest that ICErel-II and ICErel-III are also synthesized as larger proenzymes which are proteolytically processed to form heterodimeric active enzymes. Pro-interleukin-1 beta processing activity could not be detected in cells transfected with ICErel-II or ICErel-III, but pro-domain-less truncated forms of ICErel-II and ICErel-III were capable of effectively inducing fibroblast apoptosis. ICErel-II and ICErel-III may, therefore, participate in proteolytic events culminating in the apoptotic death of human cells.

Amino Acid Sequence↗

Epidemiology of smallpox in west Pakistan. III. Outbreak detection and interlocality transmission. 1971.

During one year, 121 outbreaks of variola major were detected in 99 of the 1717 localities within a rural area of West Pakistan with a total population of approximately 1.2 million. Only 19% of the outbreaks, representing 36% of the 1040 investigated cases, were officially reported, although potential strengths in the government surveillance system were also identified. Persistence of smallpox within the area depended on introductions from the outside, and more than one-half of all outbreaks of known source could be ultimately traced to cities. Within the study area, outbreaks with the largest numbers of cases and those in the larger communities were the ones from which smallpox was most frequently transported. The frequency with which variola was introduced into localities was directly related to population size and to the presence of medical care facilities. Trips between localities by infected individuals were extremely rare events. They were made most often during the late fall and winter, primarily during the incubation period of the disease, and did not differ in purpose, means or destination from journeys unassociated with smallpox. Individuals at relatively high risk of becoming introducers included the unvaccinated (primarily children under five), the unschooled and those not native to the area. Vaccination priorities based on these findings could increase the efficiency of smallpox eradication efforts.

Disease Outbreaks↗

Seasonal variation in follicular dynamics of superovulated Indian water buffalo.

The ovaries of 5 buffalo were examined daily by ultrasound beginning at Day 3 of the estrous cycle, followed by superovulation between Days 11 and 13 of the cycle in both the wet cool and dry hot seasons. Daily ultrasonographic observations of the ovaries were recorded on a videotape and were used to assess the progression of both the large (dominant) and the next to the large (sub-dominant) follicles as well as the numbers of follicles in the small (4 to 6 mm), medium (7 to 10 mm) and large (> 10 mm) size categories in the 2 seasons before and during the superovulation treatment. Greater numbers of small (P < 0.05) and medium size follicles (P < 0.01) were available before the start of the superovulatory treatment in the buffalo during the dry hot season. The turnover of follicles from medium to large size classes also occurred sooner (P < 0.01) and was of higher magnitude (P < 0.05) during the treatment in the dry hot season. However, the number of corpora lutea at palpation per rectum (2.8 +/- 0.7 vs 2.2 +/- 0.6), the serum progesterone concentration (1.6 +/- 0.3 vs 1.4 +/- 0.1 ng/ml), and the yield of embryos on Day 6 (0.2 +/- 0.2 vs 0.6 +/- 0.2) did not differ significantly between the dry hot and the wet cool season. None of the embryos recovered during the dry hot season were transferable, which remains unexplained.

Journal Article↗

Characterization of two Japanese encephalitis virus strains isolated in Thailand.

Two strains of Japanese encephalitis (JE) virus were isolated from a pool of Culex tritaeniorhynchus captured in 1992 and another pool of Cx. vishnui captured in 1993, in Chiang Mai Area, Northern Thailand. These two strains, ThCMAr44/92 and ThCMAr67/93, could not be identified either as Nakayama or JaGAr01 subtype by the hemagglutination-inhibition (HI) and the neutralization (N) tests using immune sera raised against these standard JE virus strains. Reverse transcription-polymerase chain reaction showed the presence of JE-specific conserved sequences in these strains. Sequencing of 240 nucleotides in their PrM gene region identified that these two strains belong to the genotype 1 of JE virus. Nucleotide and encoded amino acid sequences of their envelope glycoprotein gene revealed 98.8 and 99.8% identity, respectively. These two strains shared 77.8 to 87.7% homology in the nucleotide sequence and 90.0 to 98.8% homology in the amino acid sequence with other reported JE strains. Five strain-specific amino acid changes were noted in ThCMAr44/92 strain, while one in ThCMAr67/93. In addition, four common amino acid changes were found in both strains. Thus, the findings indicated that these two strains were structurally different from each other as well as different from all the reported strains which was in agreement with the serological tests by hemagglutination-inhibition and neutralization.

Animals↗

Follicular dynamics in water buffalo superovulated in presence or absence of a dominant follicle.

The ovaries of 12 buffalo were examined daily by ultrasound beginning at Day 3 of the estrous cycle, followed by superovulation between Days 10 and 13 of the cycle. The buffalo were divided into 2 groups on the basis of the presence (dominant, n = 7) or absence (nondominant, n = 5) of a dominant follicle at the start of superovulation. Daily ultrasonographic observations of the ovaries were recorded on a videotape and were used to assess the progression of both the large (dominant) follicle and the next-to-the-large (subdominant) follicle as well as the numbers of follicles in the small (4 to 6 mm), medium (7 to 10 mm), and large (>10 mm) size categories, before and during the superovulation treatment. A greater number of small size (P < 0.05) follicles was available before the start of the superovulatory treatment in the buffalo superovulated in the absence of a dominant follicle. The turnover of follicles from medium to large size classes also occurred sooner (P < 0.01), and was of higher magnitude (P < 0.01) during treatment in buffalo of the nondominant follicle group. The number of corpora lutea at palpation per rectum was higher (P < 0.05) in buffalo of the nondominant than the dominant group (4.6 +/- 0.6 vs 2.7 +/- 0.5). However, there was no significant difference among the groups in the means of serum progesterone concentration (3.6 +/- 1.3 vs 2.2 +/- 0.6 ng/ml), total number of embryos (2.0 +/- 0.6 vs 1.1 +/- 0.7), transferable embryos (1.6 +/- 0.5 vs 1.0 +/- 0.6) and unfertilized ova recovered (0.4 +/- 0.2 vs 0) on Day 6. It is concluded that in buffalo, the superovulatory response could possibly be improved by ultrasongraphic observation of the status of follicular dominance prior to treatment.

Journal Article↗

Native DNA fragments photocrosslinked to psoralen binds to anti-B and anti-Z DNA antibodies.

Calf thymus DNA fragments of varying size (100-600 bp) were covalently linked to psoralen in the presence of 365 nm UV light. The crosslinked species were separated from the monoadducts by hydroxyapatite column chromatography. The DNA-psoralen crosslink showed binding with naturally occurring anti-DNA autoantibodies. This was substantiated by the recognition of affinity purified IgG by DNA-psoralen crosslinks of varying DNA size. The photo-crosslinks exhibited considerable recognition of both monoclonal and polyclonal anti-Z-DNA antibodies in inhibition ELISA and band shift assay.

Animals↗

Maxillectomy--to reconstruct or obturate? Results of a UK survey of oral and maxillofacial surgeons.

Post-maxillectomy defects may be restored either by surgical reconstruction or by prostheses and there is continuing controversy about the most appropriate method of rehabilitation in any particular case. A questionnaire was designed to assess the current practices of oral and maxillofacial surgeons in the UK after resection of the maxilla for malignant disease. Maxillectomies were carried out by 83% of surgeons; most surgeons do 1-5 cases a year; 38% of surgeons do reconstruct surgically, but only in 10% of cases. Only 65% of surgeons have access to the services of a restorative dentist; this did influence 19% of surgeons' decision about whether to reconstruct surgically or restore by prosthetic means.

Humans↗

New insights into hepatitis C virus infection of hemodialysis patients: the implications.

The authors compared the diagnostic performance of a second-generation recombinant immunoblot assay (RIBA) (RIBA HCV 2.0 SIA) and the recently introduced third-generation RIBA (RIBA HCV 3.0 SIA) with that of hepatitis C virus (HCV) RNA by the polymerase chain reaction (PCR) in 55 patients on chronic hemodialysis. Compared with HCV RNA by PCR, RIBA 3.0 increased the sensitivity of HCV detection to 72% as compared with 56% of RIBA 2.0. Both assays underestimated the prevalence of HCV infection as determined by PCR. However, RIBA HCV 3.0 outperformed RIBA HCV 2.0, detecting all of the RIBA 2.0-positive patients plus an additional eight (8 of 22 RIBA 2.0 negative; confidence interval [CI] = [17.2%, 59.3%]). Forty-three of 51 patients with positive RIBA 3.0 or positive HCV RNA by PCR underwent a liver biopsy. Thirty (70%) had chronic hepatitis (three with cirrhosis), 10 (23%) had nonspecific changes, and three (7%) had normal liver histology. Thirty of 37 patients (81%) with hepatitis C viremia and positive anti-HCV had chronic hepatitis, whereas none of the viremic patients with negative anti-HCV had chronic hepatitis. Among the reactive antigens on RIBA 3.0, c33c was found to be most predictive of chronic hepatitis (P = 0.0002). Detection of HCV RNA continues to be the method of choice in the early phase of HCV infection. In places where a validated HCV RNA assay is not available, RIBA HCV 3.0 (soon to be commercially available) is a better alternative. Early detection of HCV infection and the implementation of an isolation strategy might be important in preventing the spread of HCV infection among hemodialysis patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Clinical performance of resin-retained fixed partial dentures bonded with a chemically active luting cement.

A clinical study was undertaken to review resin-retained fixed partial dentures bonded with a chemically active luting cement during a 4-year period. One hundred thirty-seven fixed partial dentures were reviewed of which five had failed because of a nonbonding failure (esthetics, trauma, fracture) and 47 (36%) had failed after a bonding failure, of which 72% were rebonded. The mean time of failure was 10.9 months (SD 9.8). The failure rate appeared to be affected by position, use of a rubber dam, and the fixed partial denture design, although no one factor reached conventional statistical significance.

Chi-Square Distribution↗

Comparative toxicity of selected larvicides and insect growth regulators to a Florida laboratory population of Aedes albopictus.

Five organophosphates (OPs) (chlorpyrifos, chlorpyrifos methyl, fenthion, malathion, and temephos), 3 pyrethroids (bifenthrin, cypermethrin, and permethrin), and 2 microbial pesticides (Bacillus thuringiensis serovar.israelensis [B.t.i.] and Bacillus sphaericus) were tested as larvicides against a Florida Aedes albopictus population colonized in the laboratory. In addition, 3 insect growth regulators (IGRS) (diflubenzuron, methoprene, and pyriproxyfen) were evaluated. All OPs, except for malathion, were highly effective as indicated by low LC90s ranging from 0.0069 ppm (chlorpyrifos) to 0.026 ppm (fenthion); the larvae were considered tolerant to malathion (LC90 = 1.043 ppm). LC90 values of pyrethroids were: 0.0175 ppm (bifenthrin), 0.0079 ppm (cypermethrin), and 0.0031 ppm (permethrin). Commercial products of B.t.i., Vectobac and Bactimos were considered economically effective against Ae. albopictus larvae but products of B. sphaericus were ineffective (LC90s > 28 ppm). The IGRs showed exceptional activity. Pyriproxyfen (LC90 = 0.000376 ppm), was 2.23 and 21.5 times more toxic than diflubenzuron and methoprene, respectively. In general, toxicity ranking of chemicals and microbials tested was: IGRs > pyrethroids > OPs > microbials.

Aedes↗

Quantitative myocardial SPECT for infarct sizing: feasibility of a multicenter trial evaluated using a cardiac phantom.

UNLABELLED: This study determined the feasibility of performing a multicenter trial using quantitative SPECT myocardial perfusion imaging in patients with acute myocardial infarction. The feasibility was assessed by a cardiac phantom. METHODS: Twenty-two gamma camera systems in 19 laboratories were evaluated. Each laboratory performed nine studies on the cardiac phantom and performed quality control tests of system uniformity, collimator quality and gantry alignment on their gamma camera system. Defects simulating "hypoperfused" myocardium of differing amounts were placed in the myocardium for eight of the nine studies. Measured defect size was compared to true defect size. RESULTS: A total of 198 studies from 22 systems were analyzed. Three studies were technically inadequate. For all 22 systems, the average correlation coefficient between true and measured defect size was 0.992 +/- 0.009, with a range from 1.00 to 0.97. Three systems were rejected due to slopes of the regression line outside the limits 1.00 +/- 0.10 and mean errors > 5% in estimating defect size. The remaining systems had a correlation coefficient of 0.995 + 0.008 with an average slope of 1.00 +/- 0.04 and an intercept of 0.11% +/- 1.57%. The mean error in estimating defect size was 2.08% +/- 0.69%. CONCLUSION: The small interlaboratory variation and the close correlation with true defect size observed in a cardiac phantom indicate the feasibility of quantitative myocardial SPECT as a useful tool in multicenter trials evaluating therapy in acute myocardial infarction. Preliminary objective testing is required, however, to identify systems with technical deficiencies.

Feasibility Studies↗

Characterization of the Streptomyces peucetius ATCC 29050 genes encoding doxorubicin polyketide synthase.

The dps genes of Streptomyces peucetius, encoding daunorubicin (DNR)-doxorubicin (DXR) polyketide synthase (PKS), are largely within an 8.7-kb region of DNA that has been characterized by Southern analysis, and gene sequencing, mutagenesis and expression experiments. This region contains nine ORFs, many of whose predicted products are homologous to known PKS enzymes. Surprisingly, the gene encoding the DXR PKS acyl carrier protein is not in this region, but is located about 10 kb distant from the position it usually occupies in other gene clusters encoding type-II PKS. An in-frame deletion in the dpsB gene, encoding a putative subunit of the DXR PKS, resulted in loss of production of DXR and the known intermediates of its biosynthetic pathway, confirming that this gene and, by implication, the adjacent dps genes are required for DXR biosynthesis. This was verified by expression of the dps genes in the heterologous host, Streptomyces lividans, which resulted in the production of aklanonic acid, an early intermediate of DXR biosynthesis.

Amino Acid Sequence↗

Activation of protein kinase C down-regulates leukotriene C4 synthase activity and attenuates cysteinyl leukotriene production in an eosinophilic substrain of HL-60 cells.

An eosinophilic substrain of HL-60 cells (HL-60#7) predominantly synthesized cysteinyl leukotrienes after stimulation with the calcium ionophore A23187. Activation of protein kinase C (PKC) by phorbol 12-myristate 13-acetate (PMA) specifically attenuated cysteinyl leukotriene production without affecting the biosynthesis of non-cysteinyl leukotrienes. The inhibition of cysteinyl leukotriene biosynthesis was prevented only by specific PKC inhibitors (staurosporine and bisindolylmaleimide) but not by inhibitors of tyrosine kinases (genistein, tyrphostin 47, and herbimycin A), protein kinase A (KT5720), or the oxidative burst (apocynin). Similar results were obtained when LTC4 synthase enzymatic activity was measured directly in the presence of saturating concentrations of exogenously added substrates. Therefore, the inhibitory effects of PKC activation on cysteinyl leukotriene formation in intact cells was attributable to effects on the LTC4 synthase enzyme. The mechanism of inhibition of LTC4 synthase by PKC activation was determined by kinetic analysis to be noncompetitive in both eosinophil-like HL-60#7 cells and monocytic THP-1 cells. Contrary to the effect of PKC activation on cysteinyl leukotriene biosynthesis, the formation of prostaglandin E2 and thromboxane B2 was elevated twofold to threefold after PMA treatment, which was prevented by the PKC inhibitor, staurosporine. We propose a regulatory model in which PKC activation shifts the profile of eicosanoid mediators produced by eosinophils from cysteinyl leukotrienes to prostanoids.

Alkaloids↗