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1.
Eur J Clin Microbiol Infect Dis ; 36(4): 611-617, 2017 Apr.
Article in English | MEDLINE | ID: mdl-27837326

ABSTRACT

The correlation between hip replacement (Hip-Repl) and chronic osteomyelitis (COM) has not been studied in Asian populations. Thus, we assessed Hip-Repl-related risk of developing COM via a population-based, nationwide, retrospective cohort study. The Hip-Repl cohort was obtained from Taiwan's Longitudinal Health Insurance Database 2000, and included patients who underwent Hip-Repl between 2000 and 2010; the control cohort was also selected from this database. Patients with a history of COM were excluded in both cohorts. We used univariate and multivariate Cox proportional hazards regression models to calculate the adjusted hazard ratios (aHRs) by age, sex, and comorbidities for developing COM. A total of 5349 patients who received a Hip-Repl and 10,372 matched controls were enrolled. In the Hip-Repl group, the risk for COM was 4.18-fold [95 % confidence interval (CI) = 2.24-7.80] higher than that in the control group after adjustment. For patients aged ≤65 years, the risk was 10.0-fold higher (95 % CI = 2.89-34.6). Furthermore, the risk was higher in the Hip-Repl cohort than in the non-Hip-Repl cohort, for both patients without comorbidity (aHR = 16.5, 95 % CI = 2.07-132.3) and those with comorbidity (aHR = 3.49, 95 % CI = 1.78-6.83). The impact of Hip-Repl on the risk for COM was greater among patients not using immunosuppressive drugs, and occurred during the first postoperative year. Patients who received Hip-Repl have an increased risk of developing COM. This risk was higher among males and patients aged 65 years or younger, and during the first postoperative year.


Subject(s)
Arthroplasty, Replacement, Hip/adverse effects , Osteomyelitis/epidemiology , Adult , Age Factors , Aged , Aged, 80 and over , Asian People , Case-Control Studies , Chronic Disease , Female , Humans , Male , Middle Aged , Risk Assessment , Sex Factors , Taiwan/epidemiology , Young Adult
2.
Eur J Clin Microbiol Infect Dis ; 34(3): 479-85, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25260789

ABSTRACT

The objective of this study was to evaluate the association between the use of anti-tuberculosis (anti-TB) agents, isoniazid (INH), rifampicin (RIF), and their combination (INH + RIF), and the risk of hepatocellular carcinoma (HCC) in cirrhotic patients. This population-based case-control study was conducted using a research database of Taiwan's National Health Insurance program. Cirrhotic patients first diagnosed with HCC between 1996 and 2011 (n = 50,351), among whom 4,738 were anti-TB medication users, were evaluated. Cirrhotic patients who did not develop HCC within the same period, frequency-matched according to age, sex, and index year, were evaluated as the control group (n = 47,488). The adjusted odds ratio (OR) of HCC was 1.34 [95 % confidence interval (CI), 1.20-1.50] in INH + RIF users compared with non-INH + RIF users. Long-term (>12 months) use of INH, RIF, and INH + RIF was significantly associated with increased risk of HCC, with an adjusted OR of 3.51 (95 % CI, 2.11-5.84), 4.17 (95 % CI, 2.76-4.31), and 7.17 (95 % CI, 4.08-12.6), respectively, after adjusting for age, sex, and comorbidities. An average dose of INH + RIF >16,050 mg/year was associated with increased risk of HCC in cirrhotic patients, with an adjusted OR of 1.48 (95 % CI, 1.27-1.73). Our results indicate that cirrhotic patients with long-term or high-dose INH and RIF treatment, particularly their combination, are associated with increased risk of HCC development.


Subject(s)
Antitubercular Agents/therapeutic use , Carcinoma, Hepatocellular/epidemiology , Liver Cirrhosis/complications , Tuberculosis/complications , Tuberculosis/drug therapy , Adult , Aged , Aged, 80 and over , Antitubercular Agents/adverse effects , Case-Control Studies , Drug Therapy, Combination/adverse effects , Drug Therapy, Combination/methods , Female , Humans , Isoniazid/adverse effects , Isoniazid/therapeutic use , Male , Middle Aged , Rifampin/adverse effects , Rifampin/therapeutic use , Risk , Risk Assessment , Taiwan , Young Adult
3.
J Frailty Aging ; 12(4): 277-283, 2023.
Article in English | MEDLINE | ID: mdl-38008977

ABSTRACT

BACKGROUND: Poor oral health is known to be associated with adverse outcomes, but the frequency and impact of poor oral health on older adults in the acute inpatient setting has been less well studied. OBJECTIVES: We examined the association between oral health, frailty, nutrition and functional decline in hospitalized older adults. DESIGN: Retrospective cross-sectional study. SETTING AND PARTICIPANTS: We included data from 465 inpatients (mean age 79.2±8.3 years) admitted acutely to a tertiary hospital. METHODS: We evaluated oral health using the Revised Oral Assessment Guide (ROAG), frailty using the Clinical Frailty Scale (CFS), malnutrition risk using the Nutritional Screening Tool (NST) and functional status using a modified Katz Activities of Daily Living (ADL) scale. We examined cross-sectional associations of oral health with frailty, malnutrition risk and functional decline on admission, followed by multivariate logistic regression models evaluating the association between poor oral health and the aforementioned outcomes. RESULTS: 343 (73.8%), 100 (21.5%) and 22 (4.7%) were classified as low, moderate and high risk on the ROAG, respectively. Poorer oral health was associated with greater severity of frailty, functional decline on admission and malnutrition risk. Abnormalities in ROAG domains of voice changes, swallowing difficulty, xerostomia, lips and tongue appearance were more frequently present at greater severity of frailty. Poor oral health was associated with frailty [odds ratio (OR): 1.76, 95% confidence interval (CI) 1.05-2.97; P=0.034]; malnutrition risk [OR: 2.76, 95% CI 1.46-5.19, P=0.002] and functional decline [OR: 1.62, 95% CI 1.01-2.59, P=0.046]. CONCLUSIONS: Poor oral health is significantly associated with frailty, malnutrition risk and functional decline in older inpatients. Oral health evaluation, as part of a comprehensive geriatric assessment may be a target for interventions to improve outcomes. Further research including longitudinal outcomes and effectiveness of specific interventions targeted at oral health are warranted in older adults in the inpatient setting.


Subject(s)
Frailty , Malnutrition , Humans , Aged , Aged, 80 and over , Frailty/diagnosis , Frailty/epidemiology , Frailty/complications , Cross-Sectional Studies , Nutritional Status , Nutrition Assessment , Activities of Daily Living , Retrospective Studies , Oral Health , Malnutrition/epidemiology , Malnutrition/diagnosis , Geriatric Assessment
4.
Oncogene ; 26(11): 1517-21, 2007 Mar 08.
Article in English | MEDLINE | ID: mdl-16953220

ABSTRACT

The p53 tumor suppressor protein plays a central role in maintaining genomic integrity by occupying a nodal point in the DNA damage control pathway. Here it integrates a wide variety of signals, responding in one of several ways, that is, cell cycle arrest, senescence or programmed cell death (apoptosis). Mutations in the tumor suppressor gene tp53, which affects the key transcriptional regulatory processes in cell growth and death, occur frequently in cancer and helps explain why p53 has been called the guardian of the genome. There is a vast body of published knowledge on all aspects of p53's role in cancer. To facilitate research, it would be helpful if this information could be collected, curated and updated in a format that is easily accessible to the user community. To this end, we initiated the p53 knowledgebase project (http://p53.bii.a-star.edu.sg). The p53 knowledgebase is a user-friendly web portal incorporating visualization and analysis tools that integrates information from the published literature with other manually curated information to facilitate knowledge discovery. This includes curated information on sequence, structural, mutation, polymorphisms, protein-protein interactions, transcription factors, transcriptional targets, antibodies and post-translational modifications that involve p53. The goal is to collect and maintain all relevant data on p53 and present it in an easily accessible format that will be useful to researchers in the field.


Subject(s)
Genes, p53 , Humans , Mutation
5.
Genetics ; 177(4): 2433-44, 2007 Dec.
Article in English | MEDLINE | ID: mdl-18073439

ABSTRACT

Most agronomical traits exhibit quantitative variation, which is controlled by multiple genes and are environmentally dependent. To study the genetic variation of flowering time in Brassica napus, a DH population and its derived reconstructed F(2) population were planted in 11 field environments. The flowering time varied greatly with environments; 60% of the phenotypic variation was attributed to genetic effects. Five to 18 QTL at a statistically significant level (SL-QTL) were detected in each environment and, on average, two new SL-QTL were discovered with each added environment. Another type of QTL, micro-real QTL (MR-QTL), was detected repeatedly from at least 2 of the 11 environments; resulting in a total of 36 SL-QTL and 6 MR-QTL. Sixty-three interacting pairs of loci were found; 50% of them were involved in QTL. Hundreds of floral transition genes in Arabidopsis were aligned with the linkage map of B. napus by in silico mapping; 28% of them aligned with QTL regions and 9% were consistent with interacting loci. One locus, BnFLC10, in N10 and a QTL cluster in N16 were specific to spring- and winter-cropped environments respectively. The number of QTL, interacting loci, and aligned functional genes revealed a complex genetic network controlling flowering time in B. napus.


Subject(s)
Arabidopsis/genetics , Brassica napus/genetics , Environment , Flowers/genetics , Genome, Plant , Quantitative Trait Loci , Chromosome Mapping , Computational Biology , Crops, Agricultural , Databases, Nucleic Acid , Gene Regulatory Networks , Genetic Variation , Seasons
6.
Oncogenesis ; 6(5): e338, 2017 May 22.
Article in English | MEDLINE | ID: mdl-28530703

ABSTRACT

TRPV4 belongs to the 'Transient Receptor Potential' (TRP) superfamily. It has been identified to profoundly affect a variety of physiological processes, including nociception, heat sensation and inflammation. Unlike other TRP superfamily channels, its role in cancers are unknown until recently when we reported TRPV4 to be required for cancer cell softness that may promote breast cancer cell extravasation and metastasis. Here, we elucidated the molecular mechanisms mediated by TRPV4 in the metastatic breast cancer cells. TRPV4-mediated signaling was demonstrated to involve Ca2+-dependent activation of AKT and downregulation of E-cadherin expression, which was abolished upon TRPV4 silencing. Functionally, TRPV4-enhanced breast caner cell transendothelial migration requires AKT activity while a combination of transcriptional and post-translational regulation contributed to the TRPV4-mediated E-cadherin downregulation. Finally, mass spectrometry analysis revealed that TRPV4 is required for the expression of a network of secreted proteins involved in extracellular matrix remodeling. In conclusion, TRPV4 may regulate breast cancer metastasis by regulating cell softness through the Ca2+-dependent AKT-E-cadherin signaling axis and regulation of the expression of extracellular proteins.

7.
Cancer Res ; 54(15): 3974-8, 1994 Aug 01.
Article in English | MEDLINE | ID: mdl-8033125

ABSTRACT

The measurement of both immune complex-bound and free unbound tumor-associated antigen was evaluated independently on a panel of sera from colon cancer patients by radioimmunoassay (RIA). A monoclonal antibody (mAb 46.3) raised against secreted antigens from human colon cancer cells in vitro was utilized in the RIA. When circulating immune complexes alone were analyzed, the data demonstrated that 5 of 5 (100%) Dukes' A patients and 11 of 16 (69%) Dukes' B patients had elevated levels of immune complexes reactive with mAb 46.3. Analysis of free circulating antigens demonstrated elevated levels of mAb 46.3-reactive antigen present in 5 of 5 (100%) Dukes' A patients and 15 of 16 (95%) Dukes' B patients. However, by analyzing total reactivity, defined by combining results from RIA with free and immune complex-bound antigen, the sensitivity of detection for Dukes' B increased to 16 of 16 (100%). Total antigen levels in sera from patients with benign diseases (ulcerative colitis, Crohn's disease, adenoma) were not significantly different from normal controls. Analysis of both free and bound antigen in RIA is, therefore, a more sensitive indicator than RIA with immune complex alone. For the advanced stages of disease, only 1 of 5 (20%) Dukes' C and 0 of 5 (0%) Dukes' D sera were positive for reactive immune complexes. When the combined RIA was evaluated, 3 of 5 (60%) and 1 of 5 (20%) Dukes' C and D sera, respectively, were positive with mAb 46.3. Taken together, these results show that RIA with mAb 46.3 is a sensitive indicator for the early stages of colon cancer.


Subject(s)
Antibodies, Monoclonal/immunology , Antigen-Antibody Complex/analysis , Antigens, Neoplasm/analysis , Colon/immunology , Colonic Neoplasms/immunology , Adult , Animals , Colon/pathology , Colonic Neoplasms/pathology , Female , Humans , Mice , Mice, Inbred BALB C , Middle Aged , Neoplasm Staging , Radioimmunoassay
8.
Cancer Res ; 55(5): 1152-9, 1995 Mar 01.
Article in English | MEDLINE | ID: mdl-7532544

ABSTRACT

Hepatocellular carcinoma is characterized by changes in gene expression associated with cell growth and differentiation. Cell surface antigenic changes have also been described based on differential antibody reactivity between normal and neoplastic liver. We obtained a novel tumor-associated cDNA designated TA1 on the basis of its differential expression between hepatoma cells and normal liver. Sequence analysis predicted a 723-base pair open reading frame with the deduced amino acid sequence encoding an integral membrane protein containing multiple hydrophobic transmembrane domains. Database searches revealed TA1 as the likely rat homologue of E16, a recently cloned human cDNA associated with lymphocyte activation. Although noncoding sequences diverged significantly, the 95% conservation of the predicted proteins between species strongly suggests an important, although as yet undefined, function in normal cells. TA1 transcripts were detected in normal adult rat tissues including testes, brain, ovary, spleen, mammary gland, and uterus with the highest steady-state expression in placenta. Although no expression was detected in normal liver, all rat hepatomas examined expressed an abundant 3.2-kilobase transcript. TA1 expression was closely associated with progression in this tumor model and suggests this molecule, originally linked to cell activation, also plays a role in the malignant phenotype.


Subject(s)
DNA, Neoplasm/genetics , Liver Neoplasms, Experimental/genetics , Liver/physiology , Lymphocyte Activation/physiology , Membrane Proteins/physiology , Amino Acid Sequence , Animals , Bacteriophage lambda/genetics , Base Sequence , Blotting, Northern , Cell Transformation, Neoplastic , Cloning, Molecular , DNA Probes , DNA, Viral/genetics , Female , Gene Amplification , Gene Expression , Genomic Library , Liver/embryology , Liver/growth & development , Liver Regeneration/physiology , Male , Membrane Proteins/genetics , Molecular Sequence Data , Molecular Weight , Phenotype , Polymerase Chain Reaction , Pregnancy , RNA/analysis , RNA/genetics , RNA, Neoplasm/analysis , RNA, Neoplasm/genetics , Rats , Rats, Inbred ACI , Rats, Sprague-Dawley , Sequence Homology, Amino Acid , Tumor Cells, Cultured
9.
Cancer Res ; 56(17): 3934-40, 1996 Sep 01.
Article in English | MEDLINE | ID: mdl-8752160

ABSTRACT

TuAg.1 is a tumor-associated membrane glycoprotein first identified in rat hepatocellular carcinoma by monoclonal antibodies (mAbs) 324.5 and 324.9. This oncofetal antigen is also expressed by hepatocytes in cell culture but not normal adult hepatocytes in vivo. Affinity chromatography and preparative continuous elution slab-gel electrophoresis were used to separate TuAg.1 from co-purified actin and immunoglobulin. TuAg.1 was recovered as a series of bands Mr 82,000-90,000, which were pooled and subjected to CNBr digestion for primary amino acid sequence analysis. Computer database analysis of TuAg.1 peptide sequence revealed homology to the rat colon carcinoma-associated antigen pE4, a member of the immunoglobulin gene superfamily. Oligonucleotide primers derived from sequences shared by TuAg.1 and pE4 were used in reverse transcription-PCR to amplify tumor-specific products corresponding to TuAg.1 cDNA. Northern blot analysis with one of these products confirmed the oncofetal expression of transcripts related to TuAg.1/pE4 and indicated an RNA species of different size expressed only in normal liver. Identity between TuAg.1 and pE4 was further confirmed by immunochemical analysis with mAb 324.5 and mAb E4. Both antibodies were reactive with the same protein on transplantable hepatocellular carcinoma AS30D but recognized different epitopes. The reactivity of human tumor cells with mAb 324.5 and 324.9 indicates the presence of a related TuAg.1 molecule expressed in human neoplasia as well.


Subject(s)
Antigens, Neoplasm/analysis , Genes, Immunoglobulin , Liver Neoplasms, Experimental/chemistry , Adult , Amino Acid Sequence , Animals , Antibodies, Monoclonal , Antigens, Neoplasm/genetics , Antigens, Neoplasm/isolation & purification , Base Sequence , Biomarkers, Tumor , Blotting, Northern , Epitopes , Humans , Immunohistochemistry , Molecular Sequence Data , Polymerase Chain Reaction , RNA/analysis , RNA/genetics , Rats , Rats, Sprague-Dawley , Transcription, Genetic
10.
Neuroscience ; 324: 69-81, 2016 Jun 02.
Article in English | MEDLINE | ID: mdl-26964679

ABSTRACT

Inter-alpha Inhibitor Proteins (IAIPs) are a family of related serine protease inhibitors. IAIPs are important components of the systemic innate immune system. We have identified endogenous IAIPs in the central nervous system (CNS) of sheep during development and shown that treatment with IAIPs reduces neuronal cell death and improves behavioral outcomes in neonatal rats after hypoxic-ischemic brain injury. The presence of IAIPs in CNS along with their exogenous neuroprotective properties suggests that endogenous IAIPs could be part of the innate immune system in CNS. The purpose of this study was to characterize expression and localization of IAIPs in CNS. We examined cellular expressions of IAIPs in vitro in cultured cortical mouse neurons, in cultured rat neurons, microglia, and astrocytes, and in vivo on brain sections by immunohistochemistry from embryonic (E) day 18 mice and postnatal (P) day 10 rats. Cultured cortical mouse neurons expressed the light chain gene Ambp and heavy chain genes Itih-1, 2, 3, 4, and 5 mRNA transcripts and IAIP proteins. IAIP proteins were detected by immunohistochemistry in cultured cells as well as brain sections from E18 mice and P10 rats. Immunoreactivity was found in neurons, microglia, astrocytes and oligodendroglia in multiple brain regions including cortex and hippocampus, as well as within both the ependyma and choroid plexus. Our findings suggest that IAIPs are endogenous proteins expressed in a wide variety of cell types and regions both in vitro and in vivo in rodent CNS. We speculate that endogenous IAIPs may represent endogenous neuroprotective immunomodulatory proteins within the CNS.


Subject(s)
Alpha-Globulins/metabolism , Brain/cytology , Brain/metabolism , Animals , Astrocytes/cytology , Astrocytes/metabolism , Brain/growth & development , Cells, Cultured , Immunohistochemistry , Mice, Inbred C57BL , Microglia/cytology , Microglia/metabolism , Neurons/cytology , Neurons/metabolism , RNA, Messenger/metabolism , Rats, Wistar
11.
Cancer Lett ; 24(3): 251-5, 1984 Oct.
Article in English | MEDLINE | ID: mdl-6498804

ABSTRACT

A diet deficient in choline and methionine, known to produce hepatocellular carcinoma in the absence of any added chemical carcinogen, induced lipid peroxidation in the nuclear fraction of the liver when fed to male Fischer 344 rats. This lipid peroxidation was detected within 1 day of feeding the diet by the appearance of diene conjugates and increased progressively up to 3 days. It was prevented completely by the addition of choline chloride to the diet. The close proximity of DNA may make it a possible target for attack by free radicals.


Subject(s)
Cell Nucleus/metabolism , Choline Deficiency/metabolism , Lipid Peroxides/metabolism , Liver/metabolism , Methionine/deficiency , Animals , Free Radicals , Liver Neoplasms/etiology , Male , Rats , Rats, Inbred F344
12.
Brain Res Mol Brain Res ; 48(1): 67-72, 1997 Aug.
Article in English | MEDLINE | ID: mdl-9379851

ABSTRACT

Arginine vasopressin (AVP) plays an important role in the regulation of secretory function and hemodynamics of choroid plexus, the primary site of cerebrospinal fluid (CSF) production. In the present study, localization of AVP and its transcripts in choroid plexus of adult male Sprague-Dawley rats was studied by immunohistochemistry and in situ hybridization histochemistry, respectively. For immunohistochemical analysis, AVP-specific polyclonal rabbit antibody was employed. Plasmid, pGrVP, containing a 232-bp fragment of rat AVP cDNA encoding the C-terminus of proAVP, was used as a probe to detect AVP mRNA. AVP-immunoreactive product was predominantly localized close to the apical (CSF-facing) membrane of choroidal epithelium while AVP transcripts were distributed throughout the cytoplasm of the cells. Our findings indicate that AVP is synthesized in choroid plexus epithelium, which suggests autocrine and/or paracrine actions of this peptide in choroidal tissue.


Subject(s)
Arginine Vasopressin/analysis , Arginine Vasopressin/biosynthesis , Choroid Plexus/metabolism , Epithelial Cells/metabolism , RNA, Messenger/analysis , Animals , Antibodies , Choroid Plexus/cytology , Immunohistochemistry , In Situ Hybridization , Male , Oligonucleotide Probes , RNA, Messenger/biosynthesis , Rabbits , Rats , Rats, Sprague-Dawley , Transcription, Genetic
13.
Mol Cells ; 8(4): 466-70, 1998 Aug 31.
Article in English | MEDLINE | ID: mdl-9749535

ABSTRACT

The plastid ribosomal protein s16 (rps16) gene was cloned from potato (Solanum tuberosum L. ssp. tuberosum cv Desiree) by PCR amplification to obtain a new homologous recombination site of plastid transformation. The potato rps16 genomic clone was 1627 bp in size and the coding region was interrupted by an 859 bp intron. Exon I was 40 bp, encoding 13 amino acids and exon II was 227 bp, encoding a 76 amino acid polypeptide. The nucleotide sequence of the rps16 gene from the "Désirée" potato shared perfect identity with the sequence from the "Superior" potato in the coding region. Three nucleotide substitutions, two nucleotide insertions, and one nucleotide deletion were found between the intron sequence of both "Désirée" and "Superior" cultivars. The amino acid sequence of the potato rps16 gene showed a high level of identity with rice, maize, tobacco, and mustard (84-94%) and a relatively low level compared with Bacillus stearothermophilus and E. coli (27-28%). Expression of the rps16 gene was strong in chloroplasts and transcripts were detectable in amyloplasts, suggesting that the rps16 gene is active in nonphotosynthetic plastids as well as in photosynthetic plastids. These results indicate that the potato rps16 gene can be used as a new homologous recombination site of plastid transformation for potato cultivars.


Subject(s)
Genes, Plant/genetics , Plastids/genetics , Ribosomal Proteins/genetics , Solanum tuberosum/genetics , Amino Acid Sequence , Base Sequence , Blotting, Southern , Chloroplasts/chemistry , Chloroplasts/genetics , Cloning, Molecular , DNA, Plant/chemistry , DNA, Plant/genetics , Exons/genetics , Gene Expression , Introns/genetics , Molecular Sequence Data , Plastids/chemistry , Sequence Alignment , Sequence Analysis, DNA , Sequence Homology, Amino Acid , Solanum tuberosum/chemistry , Species Specificity
14.
J Chromatogr A ; 743(1): 163-70, 1996 Aug 30.
Article in English | MEDLINE | ID: mdl-8817879

ABSTRACT

A purification and on-line monitoring procedure for IgM was developed. Perfusion ion-exchange chromatography was used for rapid purification of IgM from ascites fluid and hybridoma supernatant. Crude ascites was directly loaded onto an ion exchanger. Due to the complexity of IgM, a two-step ion-exchange procedure had to be developed. This procedure involved a rapid cation-exchange chromatography capture step followed by further purification using anion-exchange chromatography. High linear velocities, in excess of 3500 cm/h, enabled separations to be performed under 5 min. Purity of the final product by SDS-PAGE was shown to be greater than 95%. Furthermore, the antibodies retained biological activity as measured by indirect immunofluorescence (IIF) and ELISA. The IgM peak was also monitored on-line using a novel peak tracking approach. This involved placing an antibody column (specific to the IgM) prior to the ion-exchange column and operating the ion-exchange column with and without the antibody column in-line. The missing peak that is identified by comparing the two chromatograms indicates where the IgM elutes.


Subject(s)
Ascites/immunology , Chromatography, Ion Exchange/methods , Immunoglobulin M/isolation & purification , Online Systems , Animals , Anion Exchange Resins , Antibodies, Monoclonal/immunology , Cation Exchange Resins , Electrophoresis, Polyacrylamide Gel , Enzyme-Linked Immunosorbent Assay , Female , Fluorescent Antibody Technique, Indirect , Hybridomas/cytology , Hybridomas/immunology , Mice , Mice, Inbred BALB C , Silver Staining , Sodium Dodecyl Sulfate/chemistry , Time Factors
15.
J Chromatogr A ; 803(1-2): 61-71, 1998 Apr 17.
Article in English | MEDLINE | ID: mdl-9604327

ABSTRACT

Different ligands with high molecular masses are immobilized on compact, porous separation units and used for affinity chromatography. In subsequent experiments different enzymes are immobilized and used for converting substrates with low and high molecular masses. Disk or tube with immobilized concanavalin A (ConA) are used as model systems for lectin affinity chromatography. The enzyme glucose oxidase is used as a standard protein to test the ConA units. Subsequently glycoproteins from plasma membranes of rat liver are separated, using units with immobilized ConA. The enzyme dipeptidyl peptidase i.v., which is used as a model protein in the experiments, is enriched about 40-fold in a single step, with a yield of over 90%. The results are only slightly better than those obtained with ConA when it is immobilized on bulk supports. The important improvement lies in the reduction of separation time to only 1 h. Experiments concerning the isolation of monoclonal antibodies against clotting factor VIII (FVIII) are carried out on disks, combining anion-exchange chromatography and protein A affinity chromatography as a model for multidimensional chromatography. Both IgG (bound to the protein A disk) and accompanying proteins (bound to the anion-exchange disk) from mouse ascites fluid are retarded and eluted separately. With the immobilized enzymes invertase and glucose oxidase (GOX) the corresponding substrates with low molecular masses, saccharose and glucose, are converted. It is shown that the amount of immobilized enzyme and the concentration of the substrate are responsible for the extent of the conversion, whereas the flow-rates used in the experiments have no effect at all. The influence of immobilization chemistry was investigated with GOX. Indirect immobilization with ConA as spacer proved to be the best alternative. With trypsin, immobilized on a disk, substrates with high molecular masses are digested in flow-through. For optimal digestion the proteins have to be denatured in the buffer for sodium dodecyl sulfate-polyacrlyamide gel electrophoresis prior to application. In contrast to the conversion of substrates with low molecular masses, flow-rates play an important part in conversion of substrates with high molecular masses. With lower flow-rates a higher degree of digestion is achieved.


Subject(s)
Chromatography, Affinity/methods , Concanavalin A/chemistry , Enzymes, Immobilized/analysis , Staphylococcal Protein A/chemistry , Animals , Ascites/immunology , Dipeptidyl Peptidase 4/analysis , Dipeptidyl Peptidase 4/chemistry , Dipeptidyl Peptidase 4/metabolism , Electrophoresis, Polyacrylamide Gel , Enzymes, Immobilized/chemistry , Enzymes, Immobilized/metabolism , Glucose Oxidase/analysis , Glucose Oxidase/chemistry , Glucose Oxidase/metabolism , Glycoside Hydrolases/analysis , Glycoside Hydrolases/chemistry , Glycoside Hydrolases/metabolism , Immunoglobulin G/analysis , Immunoglobulin G/chemistry , Ligands , Liver/enzymology , Mice , Molecular Weight , Rats , Sucrose/metabolism , Transferrin/metabolism , Trypsin/analysis , Trypsin/chemistry , Trypsin/metabolism , beta-Fructofuranosidase
16.
J Comp Pathol ; 148(2-3): 252-8, 2013 Feb.
Article in English | MEDLINE | ID: mdl-22819013

ABSTRACT

Cytokines have gained increasing attention as therapeutic targets in inflammation-related disorders and inflammatory conditions have been investigated in sheep. Monoclonal antibodies (mAbs) specific for the ovine pro-inflammatory cytokines interleukin (IL)-1ß and IL-6 could be used to study the effects of blocking pro-inflammatory cytokines in sheep. Ovine-specific IL-1ß and IL-6 proteins and mAbs specific for these molecules were produced and the ability of the mAbs to neutralize the proteins was tested in cultures of ovine splenic mononuclear cells. Expression of nuclear factor (NF)-κß and signal transducer and activator of transcription (STAT)-3 was evaluated by western blotting and densitometric quantification. Treatment with purified IL-1ß and IL-6 proteins increased NF-κß (P < 0.001) and STAT-3 (P < 0.01) expression, respectively, in cell culture. Treatment with these proteins that were pre-incubated with IL-1ß and IL-6 mAbs attenuated (P < 0.01) these effects. These results confirm the bioactivity of ovine IL-1ß and IL-6 proteins and the neutralizing capacity of anti-ovine-IL-1ß and -IL-6 mAbs in vitro. These mAbs could be used to investigate anti-inflammatory strategies for attenuation of the effects of these pro-inflammatory cytokines in sheep.


Subject(s)
Antibodies, Neutralizing/pharmacology , Cytokines/immunology , Cytokines/metabolism , Spleen/drug effects , Spleen/metabolism , Animals , Antibodies, Neutralizing/immunology , Cells, Cultured , In Vitro Techniques , Inflammation/drug therapy , Inflammation/metabolism , Inflammation/veterinary , Interleukin-1beta/immunology , Interleukin-1beta/metabolism , Interleukin-6/immunology , Interleukin-6/metabolism , Models, Animal , NF-kappa B/metabolism , STAT3 Transcription Factor/metabolism , Sheep , Sheep Diseases/drug therapy , Sheep Diseases/metabolism , Spleen/cytology
17.
Nepal J Ophthalmol ; 5(2): 161-8, 2013.
Article in English | MEDLINE | ID: mdl-24172549

ABSTRACT

INTRODUCTION: Computer vision syndrome (CVS) is a condition in which a person experiences one or more of eye symptoms as a result of prolonged working on a computer. OBJECTIVES: To determine the prevalence of CVS symptoms, knowledge and practices of computer use in students studying in different universities in Malaysia, and to evaluate the association of various factors in computer use with the occurrence of symptoms. MATERIAL AND METHODS: In a cross sectional, questionnaire survey study, data was collected in college students regarding the demography, use of spectacles, duration of daily continuous use of computer, symptoms of CVS, preventive measures taken to reduce the symptoms, use of radiation filter on the computer screen, and lighting in the room. RESULTS: A total of 795 students, aged between 18 and 25 years, from five universities in Malaysia were surveyed. The prevalence of symptoms of CVS (one or more) was found to be 89.9%; the most disturbing symptom was headache (19.7%) followed by eye strain (16.4%). Students who used computer for more than 2 hours per day experienced significantly more symptoms of CVS (p=0.0001). Looking at far objects in-between the work was significantly (p=0.0008) associated with less frequency of CVS symptoms. The use of radiation filter on the screen (p=0.6777) did not help in reducing the CVS symptoms. CONCLUSION: Ninety percent of university students in Malaysia experienced symptoms related to CVS, which was seen more often in those who used computer for more than 2 hours continuously per day.


Subject(s)
Asthenopia/epidemiology , Computers , Ergonomics/statistics & numerical data , Headache/epidemiology , Students/statistics & numerical data , Adolescent , Adult , Asthenopia/drug therapy , Asthenopia/etiology , Ergonomics/methods , Female , Headache/etiology , Health Knowledge, Attitudes, Practice , Humans , Malaysia/epidemiology , Male , Ophthalmic Solutions/administration & dosage , Prevalence , Surveys and Questionnaires , Syndrome , Universities , Visual Acuity , Young Adult
18.
Plant Biol (Stuttg) ; 14(3): 411-9, 2012 May.
Article in English | MEDLINE | ID: mdl-22117561

ABSTRACT

Zoysia grass and creeping bentgrass are important turf grasses used in parks, gardens and playing fields. Development of grasses with increased tiller formation will enhance their commercial cultivation. To investigate the regulatory mechanism of tiller formation, we cloned the Zoysia japonica Lateral suppressor-like (ZjLsL) gene. The Lateral suppressor (Ls) gene encodes a transcriptional regulator belonging to the plant-specific GRAS protein family of putative transcription factors, and regulates axillary meristem initiation. A full-length DNA of the ZjLsL gene was isolated by 5'/3' DNA walking. Phylogenetic analysis showed that ZjLsL is closely related to Ls genes. Southern blot analysis revealed that zoysia grass has two copies of the ZjLsL gene. ZjLsL expression was detected in all organs of zoysia grass but was most highly expressed in culms. Overexpression of ZjLsL in creeping bentgrass and Arabidopsis plants promoted axillary bud formation. These results suggest that ZjLsL plays an important role in axillary meristem initiation and tiller formation.


Subject(s)
Agrostis/growth & development , Agrostis/genetics , Arabidopsis/growth & development , Arabidopsis/genetics , Meristem/growth & development , Meristem/genetics , Cloning, Molecular , Gene Expression Regulation, Developmental , Gene Expression Regulation, Plant , Genes, Plant , Phylogeny , Plants, Genetically Modified , Transcription Factors/genetics , Transformation, Genetic
19.
Singapore Med J ; 52(3): e48-51, 2011 Mar.
Article in English | MEDLINE | ID: mdl-21451915

ABSTRACT

We report a case of combined heart and liver transplantation for familial amyloid polyneuropathy. This is the first such combined transplant performed in Asia, and differs from previously described cases, in that cardiopulmonary bypass was continued at partial flow during liver transplantation in our case. This was done in order to provide haemodynamic support to the cardiac graft and to protect it from the impending reperfusion insult that frequently accompanies liver transplantation. The utility of this management course is discussed, along with its actual and potential complications. We also describe the impact of a lung-protective ventilation strategy employed during cardiac transplantation.


Subject(s)
Cardiopulmonary Bypass/methods , Heart Transplantation/methods , Liver Transplantation/methods , Amyloid Neuropathies, Familial/therapy , Heart Failure/therapy , Hemodynamics , Humans , Liver/pathology , Liver/surgery , Liver Failure/therapy , Male , Middle Aged , Reperfusion , Treatment Outcome
20.
J Nutr Health Aging ; 14(1): 23-8, 2010 Jan.
Article in English | MEDLINE | ID: mdl-20082050

ABSTRACT

OBJECTIVE: To study the nutritional status of nursing home residents in a multi-racial Asian society and its role in predicting short-term mortality independent of functional status and comorbidities. DESIGN: Cross-sectional study with prospective collection of mortality data. SETTING: Nursing home facility in Singapore. SUBJECTS: A total of 154 patients (mean age 77 +/- 12 years, 53.2% women). METHODS: We evaluated the demographic details, Mini Nutritional Assessment (MNA) scores, body mass index (BMI) and anthropometric measurements of the participants. Functional status and comorbidities were characterized by the modified Barthel Index and Charlson's comorbidity index respectively. RESULTS: Prevalence of undernutrition were 52% (n= 80) and 39% (n=60) when determined by BMI < 18.5 kg/m2 and MNA <17 respectively. Mortality was 25.3% (n= 39) over 2 years. Baseline factors associated with mortality include increased age, low Barthel's score, BMI < 18.5 kg/m2 and MNA < 17 (OR= 1.05, 1.01, 3.08 and 3.03 respectively, all p < 0.05). The association between low BMI and mortality remained significant (p=0.027) after adjustment for patient's age, gender, Barthel's and Charlson's scores, and prior nutritional intervention, but the association between MNA and mortality was diminished (p=0.106). CONCLUSION: There was a high prevalence of undernutrition in this nursing home population, and the diagnosis is an important predictor of mortality. Formal nutritional screening and targeted interventions may improve important clinical outcomes.


Subject(s)
Geriatric Assessment/methods , Malnutrition/mortality , Nutrition Assessment , Age Factors , Aged , Anthropometry , Body Mass Index , Cross-Sectional Studies , Female , Homes for the Aged , Humans , Male , Malnutrition/diagnosis , Malnutrition/epidemiology , Nursing Homes , Nutritional Status , Prevalence , Prospective Studies , Risk Assessment , Singapore/epidemiology
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