Biomedical subjects

J Zhou

Publications and source records attributed to J Zhou.

At least 307 records · Page 17Linked to original sources

PubMed2000

Mutant prevention concentration as a measure of fluoroquinolone potency against mycobacteria.

Mutant prevention concentration (MPC) has been proposed as a new measure of antibiotic potency by which the ability to restrict selection of resistant mutants is evaluated. To determine whether MPC provides potency information unavailable from the more customary measurement of the MIC, 18 fluoroquinolones were examined for their ability to block the growth of Mycobacterium smegmatis and to select resistant mutants from wild-type populations. Both MPC and MIC were affected by changes in the moiety at the fluoroquinolone C-8 position and in alkyl groups attached to the C-7 piperazinyl ring. When eight resistant mutants, altered in the gyrase A protein, were tested with fluoroquinolones having either a methoxy or a hydrogen at the C-8 position, the MIC for the most resistant mutant correlated better with the MPC than did the MIC for wild-type cells. For C-8-fluorine derivatives, which were generally less active than the C-8-methoxy compounds but which were more active than C-8-hydrogen derivatives, the MICs for both the mutant and the wild type correlated well with the MPCs. Thus, measurement of the MICs for wild-type cells can reflect the ability of a quinolone to restrict the selection of resistance, but often it does not. With the present series of compounds, the most potent contained a C-8-methoxy and a small group attached to the C-7 ring.

Anti-Infective Agents↗

PubMed2000

Pacific Northwest marine sediments contain ammonia-oxidizing bacteria in the beta subdivision of the Proteobacteria.

The diversity of ammonia-oxidizing bacteria in aquatic sediments was studied by retrieving ammonia monooxygenase and methane monooxygenase gene sequences. Methanotrophs dominated freshwater sediments, while beta-proteobacterial ammonia oxidizers dominated marine sediments. These results suggest that gamma-proteobacteria such as Nitrosococcus oceani are minor members of marine sediment ammonia-oxidizing communities.

Ammonia↗

PubMed2000

Nitrite reductase genes (nirK and nirS) as functional markers to investigate diversity of denitrifying bacteria in pacific northwest marine sediment communities.

Genetic heterogeneity of denitrifying bacteria in sediment samples from Puget Sound and two sites on the Washington continental margin was studied by PCR approaches amplifying nirK and nirS genes. These structurally different but functionally equivalent single-copy genes coding for nitrite reductases, a key enzyme of the denitrification process, were used as a molecular marker for denitrifying bacteria. nirS sequences could be amplified from samples of both sampling sites, whereas nirK sequences were detected only in samples from the Washington margin. To assess the underlying nir gene structure, PCR products of both genes were cloned and screened by restriction fragment length polymorphism (RFLP). Rarefraction analysis revealed a high level of diversity especially for nirS clones from Puget Sound and a slightly lower level of diversity for nirK and nirS clones from the Washington margin. One group dominated within nirK clones, but no dominance and only a few redundant clones were seen between sediment samples for nirS clones in both habitats. Hybridization and sequencing confirmed that all but one of the 228 putative nirS clones were nirS with levels of nucleotide identities as low as 45.3%. Phylogenetic analysis grouped nirS clones into three distinct subclusters within the nirS gene tree which corresponded to the two habitats from which they were obtained. These sequences had little relationship to any strain with known nirS sequences or to isolates (mostly close relatives of Pseudomonas stutzeri) from the Washington margin sediment samples. nirK clones were more closely related to each other than were the nirS clones, with 78.6% and higher nucleotide identities; clones showing only weak hybridization signals were not related to known nirK sequences. All nirK clones were also grouped into a distinct cluster which could not be placed with any strain with known nirK sequences. These findings show a very high diversity of nir sequences within small samples and that these novel nir clusters, some very divergent from known sequences, are not known in cultivated denitrifiers.

Bacteria↗

PubMed2000

Identification of the major Spanish clones of penicillin-resistant pneumococci via the Internet using multilocus sequence typing.

Multilocus sequence typing was used to characterize isolates of the major Spanish clones of penicillin-resistant and multiple-antibiotic-resistant Streptococcus pneumoniae. Isolates of the multidrug-resistant Spanish serotype 23F clone and serotype variants of this clone either had identical allelic profiles or their allelic profiles differed from this typical allelic profile at only one of the seven housekeeping loci. Similarly, isolates of the Spanish serotype 6B and 14 clones and the penicillin-resistant serotype 9V clone (and serotype variants of this clone) each had the same allelic profiles or profiles that differed at a single locus. Multilocus sequence typing therefore allows resistant pneumococci to be assigned to the Spanish clones if they have the typical allelic profile of the clone or if their profiles differ from that profile at a single locus. A few resistant isolates that had allelic profiles typical of that of a Spanish clone or whose profiles differed from that of the typical profile at only a single locus possessed penicillin-binding protein pbp1a, pbp2b, or pbp2x genes that differed from those that are characteristic of the clone. In most cases these isolates could be assigned as variant members of the clone. Since almost all serotype 9V isolates have very similar genotypes, independently emerging penicillin-resistant clones of this serotype will inevitably appear to be similar by molecular typing procedures. Analysis of the pbp genes, in addition to multilocus sequence typing (or any other molecular typing procedure), is therefore required to assign isolates unambiguously to the penicillin-resistant Spanish serotype 9V clone.

Alleles↗

PubMed2000

The Est1 subunit of yeast telomerase binds the Tlc1 telomerase RNA.

Est1 is a component of yeast telomerase, and est1 mutants have senescence and telomere loss phenotypes. The exact function of Est1 is not known, and it is not homologous to components of other telomerases. We previously showed that Est1 protein coimmunoprecipitates with Tlc1 (the telomerase RNA) as well as with telomerase activity. Est1 has homology to Ebs1, an uncharacterized yeast open reading frame product, including homology to a putative RNA recognition motif (RRM) of Ebs1. Deletion of EBS1 results in short telomeres. We created point mutations in a putative RRM of Est1. One mutant was unable to complement either the senescence or the telomere loss phenotype of est1 mutants. Furthermore, the mutant protein no longer coprecipitated with the Tlc1 telomerase RNA. Mutants defective in the binding of Tlc1 RNA were nevertheless capable of binding single-stranded TG-rich DNA. Our data suggest that an important role of Est1 in the telomerase complex is to bind to the Tlc1 telomerase RNA via an RRM. Since Est1 can also bind telomeric DNA, Est1 may tether telomerase to the telomere.

Amino Acid Sequence↗

PubMed2000

Inducible polymorphic ventricular tachyarrhythmias in a transgenic mouse model with a long Q-T phenotype.

We created a mouse model with a prolonged Q-T interval and spontaneous arrhythmias by overexpressing the NH(2) terminus and first transmembrane segment (Kv1.1N206Tag) of a delayed rectifier potassium channel (LQT(+/-) mouse). Analyses were performed using whole cell recordings of cardiac myocytes, surface electrocardiography, and programmed electrical stimulation. Action potential duration (APD) was prolonged to the same extent and was more highly variable in myocytes derived from LQT(+/-) and LQT(+/+) mice than in myocytes derived from wild-type (WT) FVB mice. Under ketamine anesthesia, the Q-T interval of both LQT(+/+) and LQT(+/-) mice was comparably prolonged versus that of WT mice. Stimulation of the right ventricle using an intracardiac catheter induced polymorphic ventricular tachyarrhythmias in 50% of the LQT(+/-) mice and 36% of the LQT(+/+) mice, whereas polymorphic ventricular tachyarrhythmias were not inducible in WT mice. The analyses of LQT(+/-) and LQT(+/+) mice indicate that prolongation of the Q-T interval in LQT mice is associated with prolonged APD, increased dispersion of APD among cardiocytes, and inducibility of polymorphic ventricular tachycardia, providing the substrate for spontaneous arrhythmias in these animals.

Action Potentials↗

PubMed2000

Glucagon-like peptide-1 induces cell proliferation and pancreatic-duodenum homeobox-1 expression and increases endocrine cell mass in the pancreas of old, glucose-intolerant rats.

Glucose homeostasis in mammals is maintained by insulin secretion from the beta-cells of the islets of Langerhans. Type 2 diabetes results either from primary beta-cell failure alone and/or a failure to secrete enough insulin to overcome insulin resistance. Here, we show that continuous infusion of glucagon-like peptide-1 (7-36) (GLP-1; an insulinotropic agent), to young and old animals, had effects on the beta-cell of the pancreas other than simply on the insulin secretory apparatus. Our previous studies on a rodent model of glucose intolerance, the aging Wistar rat, show that a plateau in islet size, insulin content, and beta-cell mass is reached at 13 months, despite a continuing increase in body weight. Continuous sc infusion of GLP-1 (1.5 pM/kg x min), over 5 days, resulted in normal glucose tolerance. Our current results in both young and old rats demonstrate that treatment caused an up-regulation of pancreatic-duodenum homeobox-1 (PDX-1) expression in islets and total pancreas, induced pancreatic cell proliferation, and beta-cell neogenesis. The effects on levels of PDX-1 messenger RNA were abrogated by simultaneous infusion of Exendin (9-39), a specific antagonist of GLP-1. PDX-1 protein levels increased 4-fold in whole pancreata and 6-fold in islets in response to treatment. Beta-cell mass increased to 7.2 +/- 0.58 from 4.88 +/- 0.38 mg, treated vs. control, respectively, P < 0.02. Total pancreatic insulin content also increased from 0.55 +/- 0.02 to 1.32 +/- 0.11 microg/mg total pancreatic protein. Therefore, GLP-1 would seem to be a unique therapy that can stimulate pancreatic cell proliferation and beta-cell differentiation in the pancreas of rodents.

Aging↗

PubMed2000

The central melanocortin system can directly regulate serum insulin levels.

The central melanocortin system has been demonstrated to play a pivotal role in energy homeostasis. Genetic disruption of this system causes obesity in both humans and mice. Previous experiments have shown that centrally-administered melanocortin agonists inhibit food intake and stimulate oxygen consumption. Here we report that centrally-administered melanocortin agonists also inhibit basal insulin release, and alter glucose tolerance. Furthermore, increased plasma insulin levels occur in the young lean MC4-R knockout (MC4-RKO) mouse, and impaired insulin tolerance takes place before the onset of detectable hyperphagia or obesity. These data suggest that the central melanocortin system regulates not only energy intake and expenditure, but also processes related to energy partitioning, as indicated by effects on insulin release and peripheral insulin responsiveness. Previous studies emphasize the role of excess adipose mass in the development of tissue insulin resistance, leading to type II diabetes. The data presented here show that defects in the central control of glucose homeostasis may be an additional factor in some types of obesity-associated type II diabetes.

Animals↗

PubMed2000

Noscapine and analogues as potential chemotherapeutic agents.

Noscapine, an antitussive drug, has been previously shown to inhibit the growth of cultured tumor cells and tumors implanted in nude mice. Like some other antitumor agents, noscapine targets a cellular protein, tubulin, that is responsible for the assembly of important scaffolding polymers, microtubules. As a result, noscapine causes the aberrant assembly of the cellular machinery necessary for cell division, the mitotic spindle. This is followed by apoptotic cell death. Current evidence suggests that the antitumor activity of noscapine might lie in its initiation of apoptotic pathways. Compared with other microtubule drugs, noscapine has low toxicity and wide efficacy in animal models. For clinical use, noscapine affords simple means of administration including oral, rectal, parenteral or by simple inhalation of suitable aerosols. Thus, noscapine and its analogues are potential cost-effective chemotherapeutic agents for the treatment of human cancers.

Journal Article↗

PubMed2000

Identification of leptin-induced transcripts in the mouse hypothalamus.

Long-form leptin receptor (OB-R(L)) is a signal-transducing member of the cytokine receptor superfamily that is essential for mediating the effects of leptin on mammalian body weight homeostasis. At present, the range of transcriptional targets responsive to OB-R(L) activation, and consequently, the likely mediators of leptin action, remain undefined. In this report, we have used cDNA subtractive hybridization to identify transcripts induced by leptin in immortalized hypothalamic neurons expressing OB-R(L). Differential expression of the identified transcripts in these cells was confirmed by both array technology and Northern blotting. In situ hybridization studies indicate that these transcripts are expressed in the mouse central nervous system, including nuclei of the hypothalamus that coexpress OB-R(L). Comparative in situ analysis of slices of hypothalami generated from control and leptin-injected ob/ob mice demonstrates that a subset of the identified transcripts is induced in vivo after leptin injection. The potential role of the proteins encoded by these transcripts in mediating the effects of leptin on body weight and energy homeostasis are discussed.

Adenoviridae↗

PubMed2000

RGS domain in the amino-terminus of G protein-coupled receptor kinase 2 inhibits Gq-mediated signaling.

We have previously shown that not only G protein-coupled receptor kinase (GRK) 2, but also a catalytically inactive Lys220Trp GRK2 decreases endothelin (ET)-1-induced inositol 1,4,5-trisphosphate (IP3) formation, and demonstrated the presence of phosphorylation-independent desensitization mechanism. To clarify the role of GRK2 other than that as a kinase, we characterized an RGS (regulator of G protein signaling)-like domain in the amino-terminus of GRK2. Both GRK2(1-181) and GRK2(54-174) suppressed Ca2+ responses induced by angiotensin II (Ang II) and ET-1, and bound directly with Galphaq but not Galphas nor Galphai3 in the presence of GDP and AlF4-. These results demonstrate that GRK2 regulates Gq-mediated signaling negatively by direct interaction between its RGS domain and the transitional state of Galphaq, as well as through phosphorylation of activated receptors by its kinase domain.

Aluminum Compounds↗

PubMed2000

[Delineation of 6q deletions in breast carcinoma cell lines by fluorescence in situ hybridization].

OBJECTIVE: To detect and delineate 6q deletions in ten breast carcinoma cell lines. METHODS: Fluorescence in situ hybridization(FISH) with 37 YAC probes hybridizing from 6q12 to 6q27 and a centromeric probe of chromosome 6 as internal control. RESULTS: In five cell lines, large deletions extending from 6q12 to 6q27 were found, while a small deletion involving 6q25-6q27 was identified in one cell line. In two cell lines containing different cell populations, deletions of 6q occurred in cells with polysomy 6, but not in cells with disomy 6. In addition, an overpresentation of 6q21-q22.2 was detected in one cell line. CONCLUSION: 6q25-q27 seems to be the commonly deleted region in breast cancers and 6q deletions might be secondary changes.

Chromosome Deletion↗

PubMed2000

[One of the etiological factors of digestive tract cancers in Chinese: the missense mutation Val384Asp in the hMLH1 gene].

OBJECTIVE: To investigate the etiological role of Val384Asp in the hMLH1 gene, which may be a polymorphism in Chinese, in colorectal, gastric and esophageal cancers. METHODS: Genomic DNA extracted from normal tissues were subjected to analysis in exon 12 of the hMLH1 gene by single strand conformation polymorphism(SSCP) followed by DNA sequencing of aberrant bands in 101 colorectal, 79 gastric and 76 esophageal cancer patients; in 79 and 76 first degree relatives of gastric cancer and esophageal cancer patients respectively; and in 100 healthy control individuals. RESULTS: Six percent of Chinese healthy individuals were the carriers of Val3 84Asp in the hMLH1 gene. There were significant differences in the frequencies of Val384Asp in the hMLH1 gene between the patients with colorectal cancer at young age(<45 years) and the control individuals (P<0.05), and between the gastric cancer patients with family history, the first degree relatives and the control group (P<0.05 and P<0.01 respectively). No difference was found between the esophageal cancer patients, their relatives and healthy controls. CONCLUSION: The alleles frequency of Val384Asp in the hMLH1 gene in Chinese is three percent. This missense mutation may play a part in the etiology of colorectal and gastric cancers in Chinese.

Adaptor Proteins, Signal Transducing↗

PubMed2000

[Evaluation for biocompatibility of poly-(3-hydroxypropyl)-DL-aspartamide].

To evaluate the biocompatibility of poly-(3-hydroxypropyl)-DL-aspartamide(PHPA), we utilized the laboratory methods, including subcutaneous implantation, histological examination, blood biochemical and micronucleus test. Histological examination revealed a nonspecific foreign body reaction around the PHPA and a slight inflammation around the small blood vessels of the liver on the 13th day, but all these reactions diminished or disappeared gradually after 34 days of implantation. The structures and functions of liver and kidney were normal. The results indicate that PHPA possesses a good nature in biocompatibility.

Alanine Transaminase↗

PubMed2000

[Cloning and expression of trehalose synthase genes in Escherchia coli].

Escherichia coli trehalose synthase otsBA genes have been cloned by using transposon Mu in vivo. The cloning frequency was 1.45 x 10(-3)/Kanr transductant. Genetic complements, endonuclase digestion and partial nucleotide sequencing analysis of these clones indicated that otsBA gene was on plasmid Mud5005. Subcloning 2.87 kb DNA fragment onto expression plasmids with higher or lower copy number and transforming into E. coli recipient strain FF4050, which have otsBA genes deleted respectively, the transformed strains obtained ability of growth on medium containing 0.5 mol/L NaCl and increased trehalose synthesis under high osmotic pressure. Production and accumulation of trehalose may play an important role in crop breeding. These studies will lay an important foundation for constructing an expression vector with the otsBA genes and transformation to tobacco mediated by Agrobacterium, and obtaining ability for drought resistance and high osmotic pressure tolerance.

Cloning, Molecular↗

PubMed2000

Antineoplastic effect of intracellular expression of a single-chain antibody directed against type IV collagenase.

It has been shown that the type IV collagenase with its two subtypes, 72 kDa/ MMP-2 and 92 kDa/MMP-9, plays an important role in tumor invasion and metastasis formation that occur through a mechanism of proteolytic degradation of collagen IV in the basement membrane. One possible method to specifically inhibit the function of the targeted protein of a cell is to express intracellular antibody combining site that can block the function or prevent the expression of the targeted molecule. Accordingly, intracellular antibodies against type IV collagenase may have a therapeutic use against tumor invasion and metastasis. As described in our previous reports, an anti-type IV collagenase monoclonal antibody (3D6) was obtained using the hybridoma approach, and its functional single-chain Fv fragment (scFv) named M97 was constructed based on recombinant phage display techniques. In this study, the endoplasmic reticulum (ER)-retained scFv antibody fragment was used to inhibit the function of type IV collagenase. For expression in mammalian cells, the assembled scFv M97 gene with ER retention signal encoding 6 additional amino acid (SEKDEL) was reamplified by PCR. The amplified fragments were cloned into the pcDNA3.1 vector containing the CMV early-intermediate promoter/enhancer. The resulting plasmid was sequenced and then introduced by the lipofectamine method into PG cells, a highly metastatic human lung cancer cell line and G418-resistant cells were obtained by G418 selection. After transfection, the M97 mRNA expression was observed and the type IV collagenase expression was downregulated significantly as measured by ELISA. The biological behavior of PG cells, such as the ability of in vitro invasion of colony formation on soft agar through Matrigel, were also inhibited by scFv M97 transfection. The results indicate that intracellular antibody technology represents a novel and efficient way to selectively abrogate the activity of type IV collagenase, at least in vitro. We are presently exploring the efficacy of this approach in a xenograft model of human lung cancer.

3T3 Cells↗

PubMed2000

The association between glutathione S-transferase M1 genotype and polycyclic aromatic hydrocarbon-DNA adducts in breast tissue.

A major goal in molecular epidemiology is to identify preventable environmental risk factors and susceptible subpopulations. In a hospital-based molecular epidemiological case-control study of breast cancer, we investigated the relationship between DNA damage from exposure to polycyclic aromatic hydrocarbons (PAHs) and susceptibility attributable to inherited deletion of the xenobiotic detoxifying gene, glutathione S-transferase M1 (GSTM1). Prior to breast surgery, women (n = 227) were enrolled and interviewed and donated a blood sample. PAH-DNA adduct levels were measured by immunohistochemistry in breast tissue samples retrieved from pathology blocks, and GSTM1 genotype was determined by PCR using WBC DNA. The GSTM1 analysis included 95 cases and 87 benign breast disease controls. GSTM1 genotype was not associated with breast cancer case-control status (odds ratio = 0.73; 95% confidence interval, 0.37-1.44). However, the GSTM1 null genotype predicted PAH-DNA adduct levels in malignant (beta = 0.407; P = 0.003) and nonmalignant (beta = 0.243; P = 0.05) breast tissue from cases. This relationship was not seen in tissue from controls (beta = 0.095; P = 0.341). When tissue from controls was compared with tumor tissue from cases, there was a significant case-control difference in PAH-DNA adduct levels among women who were GSTM1 null. There was no such case-control difference among women who were homozygous or heterozygous for GSTM1. There was an interaction between GSTM1 and case-control status on adduct levels in breast tissue (P = 0.002). The results suggest that genetic susceptibility to the formation of PAH-DNA adducts in breast tissue may play a role in breast cancer development.

Adult↗

PubMed2000

[Isolation and characterization of PAOX2 mutant in Pichia pastoris].

Spontaneous Mut+ mutants of P. pastoris AOX1-defective expression strain have been isolated, they were identified as phenotypically utilized methanol to grow as wild type. The results obtained from measuring growth curve when cultivated in medium in which methanol as a sole carbon source and detecting HSA protein on SDS-PAGE confirmed that the mutants have increased ability to utilize methanol and express foreign HSA gene product. The promoter region of AOX2 gene from the mutants has been cloned by PCR amplification, and the DNA fragment is 1022bp in size. Sequencing analysis showed that there are two point mutations at positions of -529 and -255 from the translation initiation codon respectively. The mutations improved AOX-1 defective function and facilitate the foreign gene for higher expression.

Alcohol Oxidoreductases↗