Sequential staining of short oligonucleotides in polyacrylamide gels with ethidium bromide and methylene blue.
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Biomedical subjects
Publications and source records attributed to C S Lim.
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Unusual modes of presentation of carcinoma of the gallbladder have been described in the literature. However, this is probably the first reported case of carcinoma of the gallbladder presenting with acute large bowel obstruction. This paper also reviews the literature pertaining to the clinical features and current treatment of carcinoma of the gallbladder. It continues to be a discouraging disease characterised by late diagnosis and a poor outcome.
Genetic manipulation of the vasculature may offer insights into the pathogenesis of coronary artery disease and may lead to gene therapy for disorders such as restenosis after percutaneous coronary angioplasty. The goal of this study was to develop a percutaneous method for gene transfer into coronary arteries of intact animals. Liposomes were used to facilitate transfection in coronary arteries with a plasmid containing the cDNA encoding luciferase. This reporter was chosen since it is not expressed in mammalian cells, and it can be quantified using a sensitive assay (light production). Mongrel dogs were catheterized, and DNA was delivered to coronary arteries via a porous perfusion balloon system. Luciferase expression was measured 3-5 days after the procedure, when the dogs were killed. Luciferase activity in control arteries (n = 12) was no higher than average background activity. Eight of 12 transfected arteries exhibited gene expression, averaging 4.3 +/- 2.1 pg luciferase (p less than 0.01, transfected versus control arteries). In addition, the ability to transfect DNA into femoral arteries without a transfection vehicle was tested. Five dogs were subjected to surgical transfection attempts in their femoral arteries with either DNA alone or DNA plus liposomes. Luciferase was expressed in all 10 femoral arteries; those treated with DNA alone expressed 35.6 +/- 8 pg luciferase, and those treated with DNA plus liposomes expressed 42.3 +/- 14 pg luciferase (p = 0.70). These results demonstrate the use of a percutaneous catheter to achieve gene transfer and expression in coronary arteries of intact dogs and suggest that the efficiency of intra-arterial gene transfer may be similar whether or not a transfection vehicle is used.
A renal transplant patient presented with ileal perforation due to histoplasmosis 3 years after transplantation. Mesenteric lymph nodes and lungs were also affected by the disease. She was successfully treated with amphotericin B followed by ketoconazole.
Gene therapy approaches have been suggested for the treatment of cardiovascular disease. Recently, direct transfer of the gene encoding beta-galactosidase into peripheral arteries of the pig has been demonstrated. To determine whether this approach is applicable to other arterial beds and to other species, we first evaluated the use of beta-galactosidase as a marker protein in the canine model. We demonstrate that variable but substantial endogenous beta-galactosidase-like activity is induced by manipulation of canine peripheral arteries, which precludes the use of this marker protein in evaluating the efficiency of gene transfer in this model. A marker gene encoding firefly luciferase was then evaluated, and background luciferase activity was found to be low in the dog even after arterial manipulation. Using the luciferase gene, we then demonstrated lipid-mediated gene transfer directly into both coronary and peripheral arteries of the intact dog. These results indicate the feasibility of in vivo gene transfer into coronary arteries and demonstrate the use of the luciferase marker protein in quantifying recombinant protein expression following gene transfer in canine models. This simple and effective method for direct in vivo gene transfer into coronary and peripheral arteries may be applicable to the localized production of therapeutically important proteins for the treatment of cardiovascular diseases.
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The aim of the present study was to assess the response of patients to a course of instruction in plaque control as influenced by friends and relatives. The subjects in the study were 41 patients who had been referred to a dental hospital for treatment. There was a major reduction in plaque and gingivitis levels in the subjects at both the 10th and 14th weeks of the study. There was a greater proportional reduction in gingivitis levels in those subjects who had a higher number of reported discussions with their friends. The effect of discussions with members of the family was less clear. Lower values for gingivitis levels at 10 weeks were associated with a higher number of discussions with parents. However the levels of gingivitis tended to be adversely affected by discussions between the subjects and their spouses. The importance of recognising the influence of family and friendship networks needs to be understood when preventive programmes are designed, especially where high levels of self-care are sought as outcomes. Further work is in progress on a group of office workers being treated in a dental practice environment. This will allow the type of advice being obtained from the various contacts to be investigated in more detail.
A study was carried out to determine the health effects of rice husk dust in Malaysian rice millers. The study population consisted of 122 male Malay workers from three rice mills, with 42 controls of similar age, sex, ethnic group, and agricultural work background. Interviews using standardised questionnaires, physical examination, total and differential white cell counts, chest radiographs, and lung function tests were performed on each of the millers and the controls. Environmental dust monitoring was also carried out in the three rice mills. Clinical, haematological, and radiological findings suggest that a distinct clinical syndrome seems to be associated with exposure to rice husk dust. The manifestations of this "rice millers' syndrome" include acute and chronic irritant effects affecting the eyes, skin, and upper respiratory tract; allergic responses such as nasal catarrh, tightness of chest, asthma, and eosinophilia; and radiological opacities in the chest, probably representing early silicosis or extrinsic allergic alveolitis.
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In the marine mollusk Aplysia, the CCAAT/enhancer-binding protein, ApC/EBP, serves as an immediate early gene in the consolidation of long-term facilitation in the synaptic connection between the sensory and motor neurons of the gill-withdrawal reflex. To further examine the role of ApC/EBP as a molecular switch of a stable form of long-term memory, we cloned the full-length coding regions of two alternatively spliced forms, the short and long form of ApC/EBP. Overexpression of each isoform by DNA microinjection resulted in a l6-fold increase in the expression of the coinjected luciferase reporter gene driven by an ERE promoter. In addition, when we overexpressed ApC/EBP in Aplysia sensory neurons, we found that the application of a single pulse of 5-HT that normally induced only short-term facilitation now induced long-term facilitation. Conversely, when we attempted to block the synthesis of native ApC/EBP by microinjecting double-strand RNA or antisense RNA, we blocked long-term facilitation in a sequence-specific manner. These data support the idea that ApC/EBP is both necessary and sufficient to consolidate short-term memory into long-term memory. Furthermore, our results suggest that this double-strand RNA interference provides a powerful tool in the study of the genes functioning in learning and memory in Aplysia by specifically inhibiting both the constitutive and induced expression of the genes.
To elucidate the prevalence and clinical implications of antineutrophil cytoplasmic antibody (ANCA) in lupus nephritis (LN), we examined ANCA by indirect immunofluorescence and by ELISA against antilactoferrin (anti-LF) and antimyeloperoxidase (anti-MPO) antibody. To discriminate perinuclear ANCA (pANCA) with antinuclear antibody (ANA), all the ANCA-positive sera were tested again after incubating patients' sera with single-stranded (SS) and double-stranded (ds) DNA. These results were compared with clinicopathologic manifestations and clinical courses of LN. ANCA was positive in 19 (37.3%) of 51 LN patients. Among these LN patients, 3 had cytoplasmic ANCA (cANCA) and 16 had pANCA. ANCA was not found in 8 SLE patients without nephritis and 30 normal controls. The presence of ANCA, particularly pANCA, was associated with the presence of nephritis (18/51 cases vs. 0/8 cases, p < 0.05), especially with diffuse proliferative lupus nephritis, WHO class IV (17/18 cases vs. 21/31 cases, p < 0.05) as well as the presence of anti-dsDNA antibody (17/19 cases vs. 18/30 cases, p < 0.05). Patients with ANCA frequently had deterioration of renal function (3/16 vs. 0/26 cases). Anti-LF antibody was positive in 13 patients. Among those, 12 patients had nephritis. Five patients with anti-LF antibody did not have ANCA, but 7 had pANCA, and 1 had cANCA. Patients with anti-LF antibody had lower initial creatinine levels than those without it [serum creatinine (mg/dl): 0.78 (0.6-1.0) vs. 1.43 (0.5-5.0), p < 0.05]. Anti-MPO antibody was positive in only 1 patient, suggesting that MPO is a rare antigen for ANCA in LN.
A 58-year-old male patient had secondary polycythemia associated with idiopathic nephrotic syndrome. The renal biopsy revealed membranous nephropathy, and the bone marrow biopsy disclosed hypercellular marrow with mild panhyperplasia. The concentration of serum erythropoietin was 8.5 mU/ml. The erythrocytosis was characterized by an increased red cell volume (40.2 ml/kg) and normal arterial oxygen saturation. There were no associated lesions that could induce secondary polycythemia, except the biopsy-proven membranous nephropathy. He was treated with prednisolone and cyclophosphamide, and the nephrotic syndrome was partially remitted after 6 weeks. With partial remission of nephrotic syndrome, the erythrocytosis was resolved. This case illustrates the rarely reported association of the nephrotic syndrome and erythrocytosis, and the resolution of erythrocytosis with improvement of nephrotic syndrome.
Emphysematous pyelonephritis is a rare life-threatening infection of the renal parenchyma. It usually affects unilateral kidney and occurs mostly in diabetic patients. It is characterized by the presence of gas within the renal parenchyma and requires prompt diagnosis and early aggressive therapy. Bilateral emphysematous pyelonephritis is even more rare and is associated with high mortality. We describe a case of a 62-year-old diabetic woman who presented with nonketotic hyperosmolar coma and bilateral emphysematous pyelonephritis caused by Klebsiella pneumoniae. Diagnosis of bilateral emphysematous pyelonephritis was confirmed by an abdominal computed tomographic scan and microbiologic studies. Our patient was successfully treated using percutaneous catheter drainage and long-term antibiotic therapy.