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Biomedical subjects

L Yu

Publications and source records attributed to L Yu.

At least 847 records · Page 47Linked to original sources
PubMed1970

Divalent cation activation of deoxycholate-solubilized and -inactivated membrane reduced nicotinamide adenine dinucleotide oxidase of Bacillus megaterium KM.

After treating Bacillus megaterium KM membranes with 0.2% sodium deoxycholate, most of the membrane reduced nicotinamide adenine dinucleotide (NADH) oxidase was inactivated, and all of the membrane NADH-2,6 dichlorophenol indophenol oxidoreductase was solubilized. Dilution of the deoxycholate-treated membranes in the presence of divalent cations restored almost all of the original membrane NADH oxidase. The effectiveness of the divalent cation activation decreased in the order Ba(2+) > Ca(2+) > Mg(2+) > Mn(2+). After centrifugation, the deoxycholate-treated membranes at 100,000 x g for 1 hr, all of the NADH oxidase that was activated by a divalent cation was soluble. Cation-activated oxidase, however, was insoluble. The results show that 0.2% deoxycholate at least partially solubilizes the total electron chain from NADH to O(2) in an inactive from which can be reactivated by divalent cations with the formation of active, insoluble NADH oxidase.

Bacillus megaterium↗
PubMed1970

Factors affecting deoxycholate inactivation and Mg++ reactivation of Bacillus megaterium KM membrane nicotinamide adenine dinucleotide (reduced form) oxidase.

Decay of the reduced nicotinamide adenine dinucleotide oxidase of Bacillus megaterium KM membranes was prevented by storage in 10% (v/v) glycerol or 0.4% bovine serum albumin. Differential rates of solubilization of components of the oxidase system by 0.4% deoxycholate was demonstrable at 4 C. The amount of Mg(++) necessary for maximal oxidase reactivation increased with increasing amounts of deoxycholate-inactivated oxidase. Mg(++) activation of deoxycholate-inactivated oxidase was partially temperature-dependent. Maximal activation was observed at 37 C, but only partial activation took place at 4 C. A small amount of deoxycholate was required for Mg(++) activation of deoxycholate-inactivated oxidase. Two pH optima were found for Mg(++) activation of deoxycholate-inactivated oxidase, pH 5.3 and 7.3. Significant amounts of activation of the inactive oxidase occurred in the absence of Mg(++) with an optimum at pH 5.0, with essentially no Mg(++)-independent activation demonstrable at pH 7.0.

Bacillus megaterium↗
PubMed1969

Hydrogenase measurement with photochemically reduced methyl viologen.

Methyl viologen was reduced photochemically in the presence of proflavine and ethylenediaminetetraacetic acid. The reduced methyl viologen was oxidized by hydrogenase from Vibrio succinogenes. H(2) and oxidized methyl viologen were the products. Hydrogenase activity was determined by spectrophotometric measurement of the disappearance of reduced methyl viologen at 600 nm. The extinction coefficient of reduced methyl viologen was determined and is 8.25 mm(-1) x cm(-1) at 600 nm. Optimal conditions for assaying V. succinogenes hydrogenase were developed. Extracts of Escherichia coli and Desulfovibrio desulfuricans catalyzed reduced methyl viologen oxidation in the assay system, whereas Azotobacter vinelandii extracts were inactive.

Bacteria↗
PubMedDate not supplied

Production of antibodies in transgenic plants.

Plants offer a cost-effective bioreactor to produce antibodies of diverse types. Recent studies demonstrate that secretory IgA, the predominant antibody isotype of the mucosal immune system, can be made in large quantities in plants. CaroRx, the lead SIgA antibody being developed by Planet Biotechnology Inc., has demonstrated activity in pilot phase II trials versus S. mutans, the major pathogen contributing to development of dental caries. Numerous other SIgA plantibodies are in preclinical development.

Antibodies, Monoclonal↗
PubMedDate not supplied

Expression of testis specific ankyrin repeat and SOCS box-containing 17 gene.

Human ASB-17 (Ankyrin Repeat and SOCS Box-containing 17) is a recently identified gene belonging to the ASB family, isolated from testis cDNA library. Human ASB-17 is expressed exclusively in testis among 16 tissues, revealed by Northern blot. Mouse Asb-17 was shown to be expressed from the third week post birth to adult by semi-quantitative RT-PCR analysis. In situ hybridization on frozen sections demonstrated that Asb-17 is expressed in spermatogenic cells in adult mouse, but not in Leydig cell and epididymis in adult mouse. ASB-17 proteins are highly conserved in mammals including human, mouse, rat, Canis familiaris and Macaca fascicularis.

Adaptor Proteins, Signal Transducing↗
PubMedDate not supplied

Expression of Cdv-iR gene in mouse epididymis as revealed by in situ hybridization.

We have previously studied mouse Cdv (carnitine deficiency-associated gene expressed in ventricle)-1 related gene Cdv-1IR and its human counterpart CDV-1R, and revealed that mouse Cdv-1R was predominantly expressed in testis by multiple tissue northern analysis. To further localize the Cdv-1R mRNA in mouse testis and epididymis tissue, in situ hybridization study was reported in this article. In the adult mice, the Cdv-1R expression was intensively found in the epithelial cells of the caput and corpus epididymis, whereas it was moderately detected in the initial segment, and weakly in the cauda epididymis. In the seminiferous tubles of the testis, no obvious hybridization signals were observed above the background level. This Cdv-1R region-specific expression pattern in the epididimis suggests Cdv-1R may play an important role in sperm maturation. Moreover, considering the Cdv-1R has a similar expression distribution in epididymis to the OCTN2, it would appear that Cdv-1R might be involved in the carnitine pathway in the epididimis.

Animals↗
PubMedDate not supplied

Experimental replacement of the thoracic esophagus with a bioabsorbable collagen sponge scaffold supported by a silicone stent in dogs.

We manufactured an esophageal prosthesis made of a collagen sponge supported by an inner silicone tube. The collagen was derived from porcine skin and was crosslinked by dehydrothermal treatment. We implanted our prosthesis in nine dogs after resecting a 5 cm length of the thoracic esophagus. Intravenous hyperalimentation was performed for 4 weeks after the surgery. After 29 days, the inner silicone tube was removed endoscopically and oral feeding was restarted immediately. All nine dogs survived more than 1 month, and three of the nine dogs survived for more than 1 year. Endoscopically, the luminal surface of the regenerated esophagus was covered with a lustrous, smooth mucosa, but stenosis was observed in the midregion of the regenerated esophagus. Microscopically, no foreign material remained at the reconstructed site 1 month after surgery; the collagen sponge was absorbed and replaced by host tissue. Complete epithelization was observed within 3 months. Regeneration of the lamina muscularis mucosae, esophageal glands, and skeletal muscle were observed near the anastomoses. Our artificial esophagus holds promise for esophageal reconstruction in a clinical setting, even in the thorax.

Animals↗
PubMedDate not supplied

Serum proteomic features for detection of endometrial cancer.

To find new potential biomarkers for detection of endometrial cancer (EC), 70 serum samples including 40 from EC patients and 30 from normal healthy females were detected by surface-enhanced laser desorption-ionization time-of-flight mass spectrometry (SELDI-TOF-MS) using WCX2 (weak cation exchange) protein chip. Mass spectra were then assessed with three powerful data-mining tools: a tree classifier, Biomarker Wizard software, and Biomarker Patterns System. The diagnostic pattern combined with 13 potential biomarkers could differentiate EC patients from healthy persons, with a specificity of 100%, sensitivity of 92.5%, and total coincidence of 95.7%. The combination of surface-enhanced laser desorption-ionization with bioinformatics tools could help find new biomarkers and establish with high sensitivity and specificity for the detection of EC.

Adult↗
PubMedDate not supplied

Function of estrogen receptor isoforms alpha and beta in endometrial carcinoma cells.

The aim of this study was to explore how to modulate the expression of estrogen receptors (ER) alpha and beta and to verify the role of ERalpha and beta in relationship to estrogen and tamoxifen (TAM). A series of oligodeoxyribonucleotides (ODN) corresponding to regions of the ERalpha or beta was tested in human endometrial cancer cell lines (HEC-1B). The change in HEC-1B proliferation in response to 17beta-estradiol (E(2)) and TAM under the impact of antisense ODN was studied. The results of the study are as follows: 1) transfection with antisense ODN significantly inhibited ERalpha and ERbeta protein production, 2) the cells lost the ability to proliferate in response to E(2) after transfection with ERalpha antisense ODN especially at 24, 48, and 72 h. There was no obvious change in response to E(2) in HEC-1B cell lines that were transfected with ERbeta antisense ODN, and 3) after transfection with ERalpha antisense ODN, HEC-1B cells lost the ability to proliferate in response to TAM at 48 h. This inhibition was also observed after transfection with ERbeta antisense ODN at 24 h. ERalpha may be the primary receptor in the proliferation of HEC-1B cells in response to E(2). Both ERalpha and ERbeta are involved in the agonist impact of TAM on endometrial cancer cells.

Cell Proliferation↗
PubMedDate not supplied

The effect of Epstein-Barr virus gene BHRF1 expression on radioresistance of nasopharyngeal carcinoma cells.

In order to investigate the effect of the expression of Epstein-Barr virus gene BHRF1 on the apoptotic resistance of nasopharyngeal carcinoma cells to radiation, a highly expressing vector for BHRF1 was constructed and transfected into the nasopharyngeal carcinoma cell line CNE2. Then, the biologic alterations of the cells were tested after 60Co radiation. The results showed that, in the BHRF1-expressing cells, the apoptotic index was far lower than in the control groups after 60Co radiation, and cells recovered faster from the radiation, with a higher cell-proliferative rate, stronger ability of colony formation and tumor development in nude mice than that in the control groups. Given the functional homology of BHRF1 with bcl-2, our data indicate that BHRF1 expression could prohibit nasopharyngeal carcinoma cellular apoptosis caused by radiation and in this way contribute to oncogenic transformation.

Animals↗
PubMedDate not supplied

Clinical trial of a plant-derived antibody on recolonization of mutans streptococci.

OBJECTIVE: This double-blinded, placebo-controlled clinical trial tested the safety and efficacy of a topical secretory IgA antibody manufactured in tobacco plants (plantibody) in preventing recolonization of mutans streptococci (MS) in human plaque as measured by whole stimulated saliva samples. METHODS: Following a 9-day antimicrobial treatment with chlorhexidine (CHX), 56 eligible adults (enrollment salivary MS > or = 10(4) CFU/ml; no current caries) were randomized equally to a group receiving 0, 2, 4, or 6 topical applications of plantibody followed by 6, 4, 2, or 0 applications of placebo, respectively, over a 3-week period. RESULTS: Among the 54 subjects who completed the trial, the CHX regimen eliminated salivary MS in 69%. After 6 months, there were no significant differences in MS levels by number of applications, relative to placebo (p > 0.43). No adverse effects were observed. CONCLUSION: Plantibody is safe but not effective at the frequency, concentration, and number of applications used in this study.

Adult↗
PubMedDate not supplied

Antitumor and antitumor-promoting actions of macrophages and their relationship with estrogen.

Silica provides, in fact, a remarkable example of selective toxicity for macrophage by a substance of simple chemical composition and low chemical reactivity. Intraperitoneal injection of silica resulted in an increase of growth rate of subcutaneously implanted mouse sarcoma 180 (S180) in female BALB/c mice and of tumor incidence of two-stage mouse skin carcinogenesis in female ICR mice, but did not show any significant effects on tumor growth and carcinogenesis in male mice. In contrast, local administration of homologous macrophages led to the decrease of growth rate of subcutaneously implanted mouse S180 in female BALB/c mice and of tumor incidence of two-stage mouse skin carcinogenesis in female ICR mice. On the other hand, estradiol, estradiol plus macrophages or a large number of macrophages alone inhibited the tumor growth in male mice. It should be noted that estradiol plus macrophages had the most potent antitumor action among them. These results suggest that macrophages, especially estrogen-activated macrophages, may play an important role in antitumor and antitumor-promoting actions of organisms.

Animals↗
PubMedDate not supplied

Growth kinetic study of human hepatocellular carcinoma using proliferating cell nuclear antigen and Lewis Y antigen: their correlation with transforming growth factor-alpha and beta 1.

The factors which control the balance between proliferation and cell death in hepatocellular carcinoma (HCC) remain unclear. The kinetic state of the tumor growth was investigated in the present study with references to transforming growth factor (TGF)-alpha and -beta 1 in 50 resected HCCs without preceding therapies. three-micrometer sections were cut from formalin-fixed, paraffin-embedded tumors. Proliferating cell nuclear antigen (PCNA), Lewis Y antigen (LeY). TGF-alpha, and -beta 1 were immunohistochemically stained and quantitatively assessed with an image analyzer. By means of immunohistochemical staining, a reciprocal correlation was observed between PCNA and LeY. A similar pattern was found between TGF-alpha and -beta 1, although not so strikingly as in the case of PCNA and LeY. The expression of LeY and PCNA labeling index (LI) ranged from 0 to 56.1% and from 0 to 52.8%, respectively. A correlation was observed between LeY and tumor size (r = 0.302, p < 0.04), while there was no significant relationship between PCNA LI and tumor size (r = -0.048, p > 0.05). The positive area ranged from 0 to 61.6% for TGF-alpha, and from 0.6 to 71.5% for TGF-beta 1. Analysis of these data showed a significant correlation between TGF-beta 1 and tumor size (r = -0.327, p < 0.03). Among HCCs < 5 cm, PCNA LI positively correlated with tumor size (r = 0.399, p < 0.04), but negatively with TGF-beta 1 (r = -0.431, p < 0.03). In conclusion, the present investigation indicates that the simultaneous analyses of proliferating and apoptotic activities by means of PCNA and LeY could yield more accurate data to determine the kinetic state of the tumor growth compared with either alone. In addition, TGF-beta 1 might act as a suppressive factor in the growth of HCC < 5 cm.

Adult↗
PubMedDate not supplied

Cloning and expression of the VP2 gene of an infectious bursal disease virus.

A serotype I infectious bursal disease virus (IBDV) strain HZ96 was isolated in Hangzhou, China, in 1996 and attenuated by adaptation to chicken embryo fibroblast cells. The VP2 gene of strain HZ96 was amplified by reverse transcription-polymerase chain reaction, cloned, and sequenced. Compared with the VP2 sequences of other IBDV strains, HZ96 is most related to two attenuated strains, CU-1 and PBG98, and two attenuated Chinese strains, Harbin and CJ801bkf. HZ96 shares nucleotide sequence homology 98.9% with CU-1 and PBG98, 98.5% with Harbin, and 98.6% with CJ801bkf. Most of the sequence variations observed between HZ96 and other strains are located in the middle variable region from nucleotides 637 to 996. Similar to other attenuated IBDVs, HZ96 has unique substitutions at residues 279 (Asp to Asn) and 284 (Ala to Thr), suggesting that these two substitutions may be directly related to adaptation of the virus to cell culture and attenuation of its virulence. As part of our effort to develop a submit vaccine for IBDV, the VP2 gene of HZ96 was cloned into a heat-inducible expression vector and expressed in Escherichia coli system. A protein band with expected molecular weight of 52 kD was detected by direct protein staining and western blotting.

Amino Acid Sequence↗
PubMedDate not supplied

Molecular epidemiology of infectious bronchitis virus isolates from China and Southeast Asia.

In order to trace the origin and evolution of avian infectious bronchitis virus (IBV) isolates in China and Southeast Asia, genomic sequencing was used for molecular characterization of 24 IBV isolates and two reference strains in comparison with the published sequences. The 5' region of the S1 genes, containing hypervariable regions I and II, and 3' region of the nucleocapsid genes, containing cytotoxic T lymphocyte epitopes, were used to construct phylogenetic trees for analysis. The results showed that the 24 isolates could be divided into three distinct groups, that is, American, Asian, and European. Some isolates formed a distinct Asian phylogenetic group, suggesting that IBV has existed for some time in Asia. Our results also showed that in vivo recombination of IBV may have occurred at a rather high frequency, contributing to the diversity of these IBV isolates. Importantly, recombination events have probably occurred between vaccine strains and field strains in the natural condition.

Animals↗
PubMedDate not supplied

Study of protection by recombinant fowl poxvirus expressing C-terminal nucleocapsid protein of infectious bronchitis virus against challenge.

A stable recombinant fowl poxvirus (rFPV) expressing the C-terminal region (119 amino acids) of the nucleocapsid (N) protein of an infectious bronchitis virus (IBV) strain Ch3 was constructed by inserting the coding sequence within the thymidine kinase gene of fowl poxvirus (FPV) by homologous recombination. The N protein was expressed under control of the vaccinia virus promoter P7.5 in chicken embryo fibroblast cell cultures as seen in immunofluorescence assay and in rFPV-inoculated specific-pathogen-free (SPF) chickens by detecting antibodies with enzyme-linked immunosorbent assay (ELISA). A homologous IBV strain (Ch3) and two heterologous IBV strains (Ch5 and H4) were used to inoculate SPF chickens in a challenge to examine the protective efficacy of the rFPV. When the chickens were challenged with IBV Ch3 or Ch5, the control birds had respiratory signs of infections bronchitis, whereas all the vaccinated birds were clinically normal although low levels of the IBV infection were detected by a differential ELISA. In contrast, in the chickens challenged with IBV H4, all control birds and vaccinated birds suffered from the highly lethal IBV H4 infection. Our results suggest that the C-terminal 119 amino acid of the nucleocapsid expressed by FPV is a host-protective antigen and may induce cross-protective immunity against illness among some IBV strains.

Amino Acid Sequence↗
PubMedDate not supplied

Characterization of three infectious bronchitis virus isolates from China associated with proventriculus in vaccinated chickens.

Outbreaks of an avian disease in infectious bronchitis-vaccinated chickens in China have led to the characterization of coronaviral isolates Q1, J2, and T3, which were isolated from proventricular tissues of the affected young layer flocks. Serologic analysis revealed that they could induce high titers of infectious bronchitis virus (IBV) antibodies in inoculated specific-pathogen-free (SPF) chickens in indirect enzyme-linked immunosorbent assay but were not neutralized by antisera specific to the IBV serotype M41 and the Australian T strain. In a pathogenicity experiment, the clinical signs and related gross lesions resembling those of field outbreaks were reproduced in SPF chickens, and viruses were reisolated from the damaged tissues, including trachea, proventriculus, duodenum, and cecal tonsil. Sequence data demonstrated the complete S1 amino acid sequences of these isolates were almost identical despite recovery from geographically different areas in China and had 47.3%-82.3% similarity in comparison with the 47 published S1 sequences. On the basis of genotyping and limited serology, the three isolates, which were responsible for field outbreaks of the disease, might be a new IBV variant.

Animals↗