Double balloon enteroscopy: Bangladesh experience.

Double balloon enteroscopy (DBE) is a newly developed endoscopic modality for diagnosis and treatment of small bowel disorders. The aim of this study was to evaluate the diagnostic and therapeutic impact of DBE in patient with suspected small bowel disease. This was a prospective study. Sixty one double balloon enteroscopy procedures (30 antegrade 31 retrograde) were done in thirty six patients (20M/16F, mean age 40 ± 12.5 range 16-65 years ) at gastroenterology department, Sir Salimullah Medical College, Dhaka between October 2011 and September 2012. Indications for DBE included chronic abdominal pain 14 (38.9%), obscure GI bleeding 11 (30.56%), Small bowel obstruction 05 (13.89%), and chronic diarrhea 06 (16.67%). The morphologic findings were ulcerations 13 (36.11%), growth 03 (8.33%), vascular ectasia 03 (8.33%) and polyp 01 (2.78%). Therapeutic interventions were performed in one patient only. No serious complications were observed. Diagnostic yields in case of chronic abdominal pain, chronic diarrhea, obscure GI bleeding and small bowel obstruction were 50%, 66%, 63% and 40% respectively. The findings were adenocarcinoma 04 (11%), lymphoma 03 (8.4%), tuberculosis 03 (8.4%), non specific findings 05 (13.9%), IPSID 01(2.8%), Crohn's disease 01 (2.8%), vascular ectasia 03 (8.33%) and normal 16 (44.44%). DBE is well tolerated, feasible and useful technique for the diagnosis as well as treatment of small intestinal disorders.

Metabolism of collagen in bone of adjuvant induced arthritic rat.

The metabolism of collagen was examined in bones of rats rendered adjuvant arthritis and matched controls using radioactive isotopic tracer techniques. The rate of the synthesis was studied after the incorporation of tritiated labeled proline into the total bone collagen and determining the content of total hydroxyproline and estimating the specific and total activities of radioactive labeled hydroxyproline. The rate of the catabolism was examined by measuring the activities of various collagen degrading proteolytic enzymes in the bone extract and by estimating the total content of hydroxyproline excreted in the urine. The degradation of collagen was also followed by measuring the specific and total radioactivities of (3H)-hydroxyproline in the urine. When (3H)-proline was injected into the adjuvant arthritic rat, the specific and total radioactivities of (3H)-hydroxyproline in bone collagen were reduced significantly in diseased bone. The activities of various enzymes involved in the catabolism of collagen and other extracellular matrix components were appreciably elevated (about 2-3 fold) in the bone extract of arthritic rat. Similarly, the specific and total activities of (3H)-hydroxyproline in urine samples were also greatly increased in arthritic rats. In addition, the decreased content of hydroxyproline in total bone collagen was accompanied by the increased excretion of urinary hydroxyproline in adjuvant arthritic rats. The results clearly suggest that the arthritic disease induces the qualitative and quantitative changes in bone composition and causes the alteration in the metabolism of collagen in diseased tissue. These observations could therefore, explain in part, the altered response of connective tissue of bone to inflammation and arthritis.

Studies on the metabolism of glycosaminoglycans under the influence of new herbal anti-inflammatory agents.

The in vivo effect of an herbal based, non-steroidal anti-inflammatory product, salai guggal, prepared from the gum resin exudate of Boswellia serrata and its active principle "boswellic acids" on glycosaminoglycan metabolism has been studied in male albino rats. The biosynthesis of sulfated glycosaminoglycans, as evaluated by the uptake of [35S]sulfate, and the content of glycosaminoglycans were measured in specimens of skin, liver, kidney and spleen. Statistical analysis of the data obtained with respect to the boswellic acids and salai guggal were compared with those of ketoprofen. A significant reduction in glycosaminoglycan biosynthesis was observed in rats treated with all of the drugs. Glycosaminoglycan content was found to be decreased in the ketoprofen-treated group, whereas that of the boswellic acids or salai guggal treated groups remained unaltered. The catabolism of glycosaminoglycans was followed by estimating the activities of lysosomal glycohydrolases, namely beta-glucuronidase, beta-N-acetylglucosaminidase, cathepsin B1, cathepsin B2 and cathepsin D, in tissues and by estimating the urinary excretion and hexosamine and uronic acid. The degradation of glycosaminoglycans was found to be reduced markedly in all drug-treated animals as compared to controls. The potential significance of boswellic acids and salai guggal was discussed in the light of changes in the metabolism of glycosaminoglycans.

Refractory immune thrombocytopenia after heart transplantation: a case report.

Chronic immune thrombocytopenia responds to steroids and splenectomy in approximately 80% of the patients. Intravenous immunoglobulin, plasmapheresis, combination chemotherapy, and androgens may help in refractory cases. This report describes the course and management of refractory immune thrombocytopenia after heart transplantation.

Milrinone echocardiographic viability test: a pilot study.

We assessed the utility of milrinone to predict recovery of function after surgical myocardial revascularization in patients with severe baseline left ventricular systolic dysfunction caused by coronary artery disease (CAD). Prediction of viable myocardial segments that will regain function after revascularization may help in the selection of patients who will benefit from coronary artery bypass graft surgery (CABG) as well as aid in the choice of target sites for coronary revascularization. We investigated 20 consecutive patients with CAD and left ventricular ejection fraction < or = 40% who had evidence of myocardial viability by either thallium scan or dobutamine viability test and were candidates for elective CABG. Left ventricular regional wall motion and global ejection fraction were assessed by transesophageal echocardiography in the operating room. Measurements were done before and 10 minutes after milrinone infusion, and immediately after CABG. Left ventricular wall motion score was derived by means of a 12-segment model. Functional improvement for each segment was defined as a wall motion change > 1. Baseline ejection fraction was 27% +/- 5% (mean +/- SD). Ejection fraction increased to 35% +/- 5% after milrinone infusion (P < .0001) and to 36% +/- 6% after CABG (P < .0001). Post-CABG ejection fraction was significantly correlated with postmilrinone ejection fraction (r = 0.65, P < .0001). Milrinone infusion resulted in augmentation of contraction in 98 of the 209 abnormal segments (wall motion score > or = 2); 91 (92.9%) of these improved after CABG. One hundred nine of the 111 segments that showed no improvement with milrinone did not improve after revascularization (98.2%). Seventy-three segments were akinetic or dyskinetic at baseline; 46 (63.0%) of these improved with milrinone. Improvement in regional wall motion after revascularization was detected in 84.8% of the segments that improved with milrinone versus only 3.7% of the segments that did not improve with milrinone. In patients with ischemic cardiomyopathy, improvement in left ventricular function (segmental wall motion and global ejection fraction) during milrinone infusion is highly predictive of improvement after CABG.

Studies on the intracellular degradation of newly synthesized collagen in 3-methylcholanthrene induced fibrosarcoma cells.

The intracellular degradation of newly synthesized collagen was studied in both normal fibroblast and 3-methylcholanthrene induced fibrosarcoma cells. The degradation of newly synthesized collagen was examined using pulse-chase experiments and radioactive labelling techniques with [3H]-proline. The percentage of intracellular proteolysis of newly synthesized collagen was determined by measuring the formation of [3H]-hydroxyproline containing fragments in alcohol-soluble and insoluble fractions of normal cells and fibrosarcoma cells in the culture. The rate of degradation of newly formed collagen was then followed by estimating the radioactivity of [3H]-hydroxyproline at different intervals, during the chase period. The results clearly demonstrated that the percent of intracellular degradation of newly synthesized collagen was approximately three fold higher in fibrosarcoma cells than in normal fibroblast cells. The increased intracellular degradation of newly formed collagen was followed by an increase in the activity of cathepsin B and L in fibrosarcoma cells. The pulse-chase experiments indicated that the rate of degradation of newly synthesized collagen in fibrosarcoma cells is relatively greater than in normal fibroblast cells. In addition, as the labelling time increased, the formation of [3H]-hydroxyproline containing peptides in the ethanol-soluble fraction were found to be increased in both normal cells and fibrosarcoma cells, but the extent of formation was higher in fibrosarcoma cells compared to normal fibroblast cells. The results of this investigation collectively suggest that the intracellular degradation of newly synthesized collagen is enhanced in fibrosarcoma cells.

Toxicological effects of an organophosphorus pesticide (dimethoate) on urinary collagen metabolites in normal and high protein diets fed female albino rats.

The effect of an organophosphorus pesticide (dimethoate) on the urinary excretion of hydroxyproline (total, nondialysable, dialysable and free fractions) and hydroxylysylglycosides, glucosylgalactosyl hydroxylysine and galactosehydroxylysine was investigated in two groups of female albino rats fed with normal and high protein diets. In comparison to controls, dimethoate treated animals were found to excrete significantly decreased amounts of urinary hydroxyproline fractions from 7th day onwards. The excretion of total hydroxylysylglycoside in urine parallels the excretion of hydroxyproline. The urinary output of both glu-gal-hyl and gal-hyl was also appreciably lower from dimethoate treated animals. The normal ratio of glu-gal-hyl and gal-hyl found in the urine of dimethoate treated animals was discussed in light of decreased turn over of collagen in both bone and skin. The effect of dimethoate in rats fed with high protein diet was comparatively less than those fed with normal diet.

Affinity purification of hexosaminidases.

Hexosaminidases A and B were purified by affinity chromatography from normal gastric mucosa, after preliminary separation of isozymes by anion exchange chromatography. Heparin and mannosamine were coupled to Sepharose 4B and used as affinity matrices and the purified enzymes were found to be homogeneous when analysed by polyacrylamide slab gel electrophoresis. This combination of 2 novel affinity chromatographic procedures is superior to existing methods in that a final yield of over 70% could be achieved. Also, the number of steps required to obtain homogeneity are less in contrast to the conventional methods used previously.

Effects of dimethoate on collagen metabolism in rats.

The effects of three different doses of dimethoate on the collagen metabolism in the tissues of female albino rats were studied by measuring the specific and total activities of 3H-hydroxyproline in the dermal, gingival and uteral collagen fractions and in the urine. Compared to controls, the total activity of 3H-hydroxyproline in the soluble collagen and in the urine at 12 h after the administration of 3H-proline was significantly lower by 44.45 and 58.12 per cent in the higher dose (2.25 mg/100 g body weight) of dimethoate treated groups respectively. The urinary excretion of hydroxyproline and the total activity of urinary 3H-hydroxyproline measured after 28 days of injection of labelled proline were decreased by 45.56 and 32.68 per cent in higher doses of dimethoate treated animals respectively but the excretions of urinary 3H-hydroxyproline were decreased by 6.36 and 2.88 per cent in lower doses of dimethoate (0.56 mg/100 g body weight) treated animals. The results of the present investigation clearly indicate that the synthesis of collagen is decreased in the higher doses of dimethoate treated animals compared to lower doses of dimethoate treated animals. In addition, the rates of catabolism of both soluble and insoluble collagens were decreased in higher doses of dimethoate treated rats. In concludes that the lower doses of dimethoate (0.56 mg) treated rats were less affected than the higher doses of dimethoate (2.25 mg) treated rats.

Separation and evaluation of changing pattern of glycosaminoglycans in 3-methyl cholanthrene induced fibrosarcoma.

The present study evaluates the glycosaminoglycans (GAGs) pattern associated with transplantable rat fibrosarcoma induced by 3-methylcholanthrene. The quantitative analysis of fractionation of GAGs in fibrosarcoma and fetal tissues was performed by enzymatic digestion. The average value of total GAGs in fibrosarcoma and fetal tissue was found to be 4 times higher than its value in the tissue of origin. GAG content showed a steady increase from 7th day onward to 25th day. Hyaluronic acid content in tumor tissue was observed to increase markedly (8-fold) against that of the normal tissue and was equivalent to that of the fetal tissue. Chondroitin sulfate level was also increased in the tumor as well as fetal tissue. The increase in the chondroitin sulfate and hyaluronic acid contents might possibly be due to the abnormal GAG metabolism in the increased production of both sulfated and nonsulfated GAGs.

The delay phenomenon: the story unfolds.

Our previous studies have shown that when a flap is delayed, there is dilation of existing vessels within the flap not ingrowth of new vessels. The maximal anatomic effect on the arterial tree occurs at the level of the reduced-caliber "choke" anastomotic vessels that link adjacent vascular territories. To further investigate the sequence of anatomic changes that occurs during the delay phenomenon, a large series of 200 rabbits and 17 dogs underwent a flap delay procedure in either skin or muscle and the tissues were examined at postoperative periods between 1 hour and 1 year by using well-established fluorescein, angiographic, light microscopic, immunohistochemical, and electron microscopic techniques. These data in the rabbit skin consistently demonstrated an initial period of vasoconstriction that resolved within 3 hours postoperatively and was followed by an active and progressive dilation of choke vessels that was most dramatic between 48 and 72 hours. In vivo intravenous fluorescein dye testing revealed an interesting parallel in that there was a temporary barrier to the flow of fluorescein that occurred at the level of the choke vessels immediately after the flap was raised and that this temporary barrier-continued to impede the flow toward the flap tip in rabbits where flaps had been delayed for periods up to 72 hours. Thereafter, there was no obstruction to the flow of fluorescein along the flap. During this early delay period of 3 days, light microscopy revealed a decrease in vessel wall thickness associated with an increase in lumen diameter. Over the next 4 days, the luminal diameter continued to dilate to a lesser extent and the vessel wall thickened. Immunohistochemical analysis showed increased cell division, maximal between 24 and 72 hours, in all layers of the choke vessel wall. During this same postoperative interval, transmission electron microscopy revealed phenotypic changes in smooth muscle cells from contractile to synthetic cells. Hypertrophy of the smooth muscle cells was also observed. The vascular endothelium, which initially showed evidence of denudation, was restored to a healthy intact appearance within the first week after delay. When followed for longer periods, long-term studies of the delayed flap of up to 1 year demonstrated dramatically a permanent dilation of the choke vessel lumen and a thickening of the choke vessel wall. In canine studies, one rectus abdominis muscle was delayed by ligating the deep inferior epigastric artery. The time sequence of choke vessel dilation, studied by sequential angiograms in vivo, was comparable to that of the rabbit skin model. To ascertain the permanence and irreversibility of this dilation, the normal circulation of the delayed rectus abdominis muscle was restored by reanastomosing the deep inferior epigastric artery. Even after a recovery period of up to 3 months, the choke vessels remained dilated and tortuous instead of reverting to their original narrow diameters. From this work, it is suggested that the choke vessel dilation seen in the delay period is a permanent and irreversible event. It is an active process associated with both an increase (hyperplasia) and an enlargement (hypertrophy) of the cells in all layers of the choke artery wall and a resultant increase in caliber of these vessels. The time sequence for delay appears to be similar in different species and in different tissues, suggesting the possibility of a universal process for delay.

Chromatographic purification and homogeneity of extracellular acid proteinase of Aspergillus Fumigatus.

The extracellular acid proteinase from A. fumigatus was purified by employing several chromatographic procedures and a 164 fold purification with 22% yield was achieved. The homogeneity of the enzyme was confirmed by SDS-Polyacrylamide gel electrophoresis, immodiffusion and immunoelectrophoresis techniques.

Effect of a new non-steroidal anti-inflammatory agent on lysosomal stability in adjuvant induced arthritis.

The effect of new non-steroidal anti-inflammatory agents on lysosomal stability was studied by determining the activity of beta-glucuronidase, a typical lysosomal enzyme, in various sub-cellular fractions and its release from the lysosome-rich fraction. Adjuvant arthritic animals showed a significant increase in the beta-glucuronidase activity in sub-cellular fractions. The increased rate of the release of beta-glucuronidase from lysosome-rich fraction clearly suggested that arthritic syndrome caused decreased stability of the lysosomes. Administration of boswellic acids or salai-guggal to arthritic animals was found to increase the lysosomal stability by inhibiting the rate of release from lysosome-rich fraction and reducing beta-glucuronidase activity in various sub-cellular fractions. Of the two anti-inflammatory agents tested, salai-guggal was found to afford more therapeutic value than boswellic acids.

Further studies on the physico-chemical properties of Rhizopus nodosus acid lipase.

The Km value of R. nodosus acid lipase was found to be 5 X 10(-2) M and 8 X 10(-3) M with olive oil and tricaprylin respectively. The lipase hydrolyzed triglycerides better than synthetic detergents and methyl esters. When synthetic triglycerides varying in fatty acid chain length were used, maximum hydrolysis was observed with tricaprylin as the substrate. Positional specificity studies indicated a preference for primary esters. The lipase was activated by albumin, NaCl and taurocholate whereas heparin had no effect. The lipase contains a single polypeptide chain with 298 amino acid residues. Glutamic acid and isoleucine were found to be the amino and carboxy-terminal residues, respectively. By gel filtration and SDS-PAGE the molecular weight was determined to be 40,000 +/- 500. The lipase was susceptible to photooxidation in the presence of methylene blue and Rose bengal whereas PMSF and thiol-group specific reagents had no appreciable effect on the lipase activity. NBS inactivated the lipase. Tryptophan residues were found to be essential for the lipase activity.

Purification and properties of a glucoamylase fraction from the culture filtrate of Rhizopus nodosus.

A glucoamylase was isolated from the culture filtrate of Rhizopus nodosus and was separated from the acid lipase by DEAE-cellulose chromatography at pH 8.0 It was purified by Concanavalin A Sepharose 4B affinity chromatography followed by CM-Sephadex chromatography 387 fold with 30.7% yield. The homogeneity of the enzyme were confirmed by polyacrylamide gel electrophoresis and immunological studies. The different physico-chemical properties of the enzyme were studied. The molecular weight of the enzyme was found to be 71,000. Ethylenediaminetetraacetic acid had no effect on the enzyme whereas Hg2+ partially inhibited the enzyme activity. Tryptophan residues were found to be essential for the enzyme activity.

Studies on the peptide bond specificity and the essential groups of an acid proteinase from Aspergillus fumigatus.

The specificity and mode of action of an acid proteinase from A. fumigatus was studied with B-chain of insulin, angiotensin II and bradykinin. With reference to the known structure of the B-chain of insulin and angiotensin II, the major sites of action were determined. Acid proteinase of A. fumigatus hydrolyzed primarily three peptide bonds in the B-chain of insulin viz. i. His(5)-Leu(6); ii. Tyr(16)-Leu(17) and iii. Phe(24)-Phe(25) bonds. Additional cleavages of the bonds His(10)-Leu(11) and Leu(15-Tyr(16) were also noted. Primary splitting sites, Tyr(16)-Leu(17) and Phe(24)-Phe(25) were identical with those reported in the work of porcine pepsin C (EC 3.4.23.3). Hydrolysis of angiotensin II was observed at Tyr(4)-Ile(5) bond. The acid proteinase was found not to be inactivated by EDTA, DEP and PCMB. The pepsin specific inhibitors viz. DAN/Cu II and EPNP showed quite appreciable inhibition, while SDS completely inactivated this acid proteinase.

Physico-chemical properties of the acid proteinase from A. fumigatus.

The physico-chemical properties of the purified acid proteinase were investigated. This enzyme was found to hydrolyze hemoglobin maximally among the different substrates used. The maximum hydrolysis of hemoglobin was found at pH 2.8 and 45 degrees C in 30 min. Km value of 5.5 X 10(-4) M was obtained from the Lineweaver Burk plot for the hydrolysis of hemoglobin. The enzyme was found to be most stable at pH 4.9. The maximum stability of the enzyme was observed with 2 per cent casein as a stabilising agent. It was found to have a molecular weight 37,500 consisting of 298 aminoacid residues. The partial specific volume of the acid proteinase was found to be 0.734 ml/g. Leucine and alanine were the amino and carboxy terminal aminoacids of the acid proteinase respectively.

Studies on the physico-chemical properties of alhagain.

The protease isolated jawasee shrub was found to hydrolyze egg albumin, casein, haemoglobin and gelatin optimally near neutral pH. Fibrin, bovin serum albumin, skin albumin and skin mucoids were hydrolyzed at slightly alkaline pH, while skin globulins were hydrolyzed at slightly acidic pH. The enzyme had no effect of fibrous collagen. The optimum conditions for the hydrolysis of 50 mg of egg albumin were found to be 50 mg of alhagain at pH 6.0 and 45 degrees C for 30 minutes. A Km value of 4.4 X 10(-3) M was obtained from the Lineweaver-Burk plot for the hydrolysis of egg albumin. The enzyme was found to be comparatively thermostable and was most stable at pH 4.7. Ultraviolet irradiation exhibited no appreciable effect on the enzyme activity. The ultraviolet absorption spectrum of alhagain in bi-distilled water resembles those of bromelain and trypsin. The sugar-containing enzyme was found to have a molecular weight of 20,650. The enzymeconsists of 189 amino acid residues per molecule, neutral and acidic amino acids being present in high concentrations. The partial specific volume of alhagain was calculated to be 0.743 ml/g from its amino acid composition. Phenylalnine and arginine formed the amino terminal amino acids of alhagain, while aspartic acid and serine were identified as its carboxy terminal amino acids. Results are discussed with relation to other plant proteases.

Gallstone disease in a rural Bangladeshi community.

BACKGROUND: The prevalence of gallstone disease (GSD) in Bangladesh is not known. We evaluated the prevalence of GSD and its relation with certain factors in a rural community in Bangladesh. METHODS: A total of 1332 persons aged 15 years and above from two villages were invited to participate in the study; 1,058 (80%) subjects responded after three invitations. Each subject answered a questionnaire, including demographic features, and underwent an upper abdominal ultrasound examination. RESULTS: GSD (current cholelithiasis and history of cholecystectomy) was detected in 5.4% of subjects. The prevalence was higher in women (7.7%) than in men (3.3%; p=0.002) The prevalence rates increased from 0.9% to 10% (p=0.0124) from those aged <30 years to those >50 years. A larger proportion of obese subjects (25/52; 48.1%) had GSD than non-obese subjects (32/1006; 3.2%). Prevalence in low, middle and high socio-economic classes was 1.5%, 5.7% and 13.4%, respectively (p=0.000). A majority (71.9%) of subjects with GSD were asymptomatic. CONCLUSION: Approximately 5% of the Bangladeshi rural community evaluated have GSD. Higher age, female gender, presence of obesity and higher socio-economic class were associated with higher prevalence of GSD.

Effect of a new herbo-mineral hypolipidemic agent on plasma lipoprotein pattern in rat atherosclerosis.

Hyperlipidemia was induced in rats by feeding an atherogenic diet for 5 months. The effect of administration of an indigenous hypolipidemic agent, Anna Kaara Raaja Sindhooram (AKRS) on the plasma lipoprotein profile was studied in the presence and absence of dietary lipid stimuli. Hyperlipidemia produced an enormous increase in the cholesterol and triglyceride contents of the low density (LDL) and very low density (VLDL) lipoprotein fractions and reduced the level of the putative non-atherogenic high density cholesterol (HDL-C). The agarose gel electrophoretic pattern showed a decrease in alpha-lipoproteins and an increase in beta-lipoproteins in the hyperlipidemic rats. AKRS treatment for 5 months altered the lipoprotein pattern favourably by raising HDL-C and lowering LDL-C in the treated rats.