Evaluation of biliary complications on magnetic resonance cholangiopancreatography and comparison with direct cholangiography after living-donor liver transplantation.

AIM: To evaluate the imaging characteristics of biliary complications following liver transplantation on magnetic resonance cholangiopancreatography (MRCP) and its diagnostic accuracy in comparison with direct cholangiography. MATERIAL AND METHODS: In this prospective study, 34 patients being evaluated for possible biliary complications after living-donor liver transplantation (LDLT) with abnormal MRCP findings were followed up for information regarding direct cholangiography either endoscopic retrograde cholangiopancreatography (ERCP) or percutaneous transhepatic cholangiography (PTC) within 7 days of MRCP. Twenty-nine patients underwent ERCP and five patients underwent PTC. RESULTS: Compared to findings at direct cholangiography, MRCP presented 96.9% sensitivity, 96.9% positive predictive value, and 94.1% accuracy for the detection of biliary complications. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy for detection of anastomotic strictures, biliary leak, and biliary stone or sludge on MRCP was found to be 100%, 84.6%, 91.3%, 100% and 94.1%; 72.7%, 95.7%, 88.9%, 88% and 88.2%; 80%, 100%, 100%, 96.7% and 97.1%, respectively. CONCLUSION: MRCP is a reliable non-invasive technique to evaluate the biliary complications following LDLT. MRCP should be the imaging method of choice for diagnosis in this setting and direct cholangiography should be reserved for cases that need therapeutic interventions.

Hoover's sign: Clinical relevance in Neurology.

Hoover's sign was described by Dr. Charles Franklin Hoover more than 100 years back to differentiate between the organic and functional weakness of pyramidal origin. This test is usually performed in the lower limbs and is valuable when on bedside one is not sure about the nature of hemiparesis. A subject with hemiparesis of organic cause while asked to flex the hip of normal leg against resistance will not exert pressure on the hand of examiner placed under the heel on the affected side while in hysterical weakness heightened pressure will be felt on the examiner's hand. The presumed genesis of this sign could be the crossed extensor reflex or the principle of synergistic contraction. It is a useful clinical test in differentiating functional and organic paresis with moderate sensitivity (63%) and high specificity (100%), but there are some limitations which should be kept in mind while evaluating a patient.

Omics of cancer.

With the advances in modern diagnostic expertise for cancer, certain approaches allowing scanning of the complete genome and the proteome are becoming very useful for researchers. These high throughput techniques have already proven power, over traditional detection methods, in differentiating disease sub-types and identifying specific genetic events during progression of cancer. This paper introduces major branches of omics-technology and their applications in the field of cancer. It also addresses current road blocks that need to be overcome and future possibilities of these methods in oncogenic detection.

Propofol sedation during endoscopy in patients with cirrhosis, and utility of psychometric tests and critical flicker frequency in assessment of recovery from sedation.

BACKGROUND AND STUDY AIMS: Patients with cirrhosis who undergo endoscopy under sedation could be at increased risk of complications. We assessed the utility of the critical flicker frequency (CFF) in the recovery of cognitive function. PATIENTS AND METHODS: This was a prospective study in patients with cirrhosis who underwent endoscopy under sedation with propofol in a tertiary care center. The main outcome was deterioration in cognitive function as measured by the number connection test A and B (NCT-A, -B), digit symbol test (DST), serial dotting test (SDT), and line tracing test (LTT) before and 2 h after endoscopy. CFF was recorded before and then every 30 min after endoscopy for the next 2 h. RESULTS: In the 108 patients there was no deterioration in results of the psychometric tests after the endoscopy (NCT-A 65.2 ± 44.4 vs. 62.4 ± 43.6 s, P = 0.01; NCT-B 110.4 ± 34.7 vs. 109.6 ± 44.6 s, P = 0.45; DST 26.2 ±1 0.0 vs. 26.7 ± 9.9, P = 0.25; SDT 88.6 ± 47.5 vs. 84.3 ± 44.1 s, P = 0.02; LTT 116.6 ± 55.2 vs. 115.4 ± 51.3 s, P = 0.47.) Patients with minimal hepatic encephalopathy (MHE; n = 64) did not show any deterioration in cognitive function at 2 h (NCT-A 87.7 ± 45.4 vs. 84.3 ± 44.9 s, P = 0.06; NCT-B 134.8 ± 65.4 vs. 132.7 ± 58.8 s, P = 0.46; DST 21.4 ± 8.9 vs. 22.2 ± 8.8, P = 0.09; SDT 107.1 ± 53.0 vs. 102.7 ± 48.5 s, P = 0.03; and LTT 131.5 ± 62.2 vs. 129.6 ± 57.2 s, P = 0.46). There was a significant difference between CFF at baseline and at 30 min and 1 h but no difference thereafter in non-MHE patients, MHE patients, and in controls. A total of 30 patients (28 %) had CFF < 38 Hz. In these patients, CFF at 2 h did not significantly differ from baseline CFF (35.9 ± 1.5 vs. 36.1 ± 2.0 Hz; P = 0.19). A total of 10 patients (9 %) had transient hypoxemia and 18 (17 %) had hypotension during the procedure. The endoscopy was completed in all patients. CONCLUSIONS: Propofol is safe in patients with cirrhosis and the CFF is a useful tool for the assessment of recovery from sedation in these patients.

Improved nonviral cancer suicide gene therapy using survivin promoter-driven mutant Bax.

Suicide gene vectors are being developed in many laboratories as an attractive approach to cancer therapy. However, the development of these therapies is hampered by safety concerns and limitations of efficacy. The use of tumor-specific promoters, such as survivin promoter, can provide much needed specificity to target tumor cells. However, the expression levels from these promoters is often suboptimal and hence it is imperative to enhance the activity of the cytotoxic gene of interest. We tested apoptotic activity of several mutants of proapoptotic gene bax that constitutively translocate to the mitochondria and induce apoptosis. One of these mutants with deletion of serine at position S184 (S184del) was found to be most active and showed significant antitumor activity when expressed by the survivin promoter. In vitro testing shows that this vector (Sur-BaxS184del) induces cell killing in a variety of tumor cell lines of different origin with significantly higher efficacy than wild-type bax (Sur-BaxWT). The increase in cytotoxicity was a result of enhanced induction of apoptosis in tumor cells. In contrast to cytomegalovirus (CMV) promoter-driven bax (CMV-Bax), Sur-BaxS184del caused minimum toxicity in normal human dermal fibroblasts validating its specificity and safety. In a mouse tumor model (DA-3, murine breast cancer cells), we show that intratumoral injection of Sur-BaxS184del resulted in tumor growth retardation to the same level as CMV-Bax. This study highlights the effectiveness of using bax mutants in combination with survivin promoter for tumor-targeted suicide gene therapy in a nonviral vector.

Role of HIV Gp41 mediated fusion/hemifusion in bystander apoptosis.

Mechanisms of HIV-mediated CD4+ T cell loss leading to immunodeficiency are amongst the most extensively studied yet unanswered questions in HIV biology. The level of CD4+ T cell depletion in HIV infected patients far exceeds the number of infected T cells, suggesting an indirect mechanism of HIV pathogenesis termed bystander cell death. Evidence is accumulating that the HIV envelope glycoprotein (Env) is a major determinant of HIV pathogenesis and plays a critical role in bystander cell death. The complex structure and function of HIV Env makes the determination of the mechanism of Env mediated apoptosis more complex than previously thought. This review will examine the complex relationship between HIV Env phenotype, coreceptor expression and immune activation in determining HIV pathogenesis. We review data here corresponding to the role of HIV Env hemifusion activity in HIV pathogenesis and how it interplays with other AIDS associated factors such as chemokine receptor expression and immune activation.

Oxidative stress in patients with essential hypertension.

BACKGROUND: Free oxygen radicals react with membrane lipids to form lipid hydroperoxides, a destructive process known as lipid peroxidation. Lipid hydroperoxides decompose to form a variety of products including malondialdehyde, which is used as an indicator of the oxidative damage of cells and tissues. Endogenous antioxidant enzymes such as superoxide dismutase counteract the oxidative damage from oxidative stress. There is increasing evidence that free radicals are involved in the pathogenesis of hypertension by altering endothelial function. We evaluated the oxidative stress and endogenous enzymatic antioxidant status in patients with essential hypertension before and 3 months after treatment with antihypertensives. METHODS: Fifty patients with essential hypertension attending the outpatient services of the Department of Medicine, Institute of Medical Sciences, Banaras Hindu University and 20 age- and sex-matched healthy controls were studied. The serum malondialdehyde and superoxide dismutase levels were measured in patients at the time of presentation and after 3 months of antihypertensive treatment. No antioxidants were given to the patients during the period of the study. RESULTS: The mean (SD) serum malondialdehyde level was found to be significantly higher (0.33 [0.07] mmol/L) in patients with hypertension compared with controls (0.21 [0.05] mmol/L; p < 0.001). This showed a significant decrease following antihypertensive therapy (0.23 [0.06] mmol/L; p < 0.001) compared with pre-treatment values. The serum superoxide dismutase activity was significantly lower in patients (6.93 [1.35] mg protein/ml of serum) compared with controls (20.12 [3.65] mg protein/ml serum; p < 0.001) at the time of presentation and, compared with the pre-treatment values, increased significantly after 3 months of treatment (10.66 [2.91] mg protein/ml of serum; p < 0.001). CONCLUSION: Our study shows that essential hypertension is associated with increased oxidative stress and reduced antioxidant status. Adequate control of blood pressure with antihypertensive therapy decreases oxidative stress and improves the antioxidant status in these patients.

Efficacy of a home-made spacer with acute exacerbation of bronchial asthma: a randomized controlled trial.

Metered dose inhaler (MDI) with spacer is the preferred method for administration of aerosolized medications in pediatric asthma. The expense of commercial spacers limits their use and indigenous alternatives have therefore been developed. Information on the clinical efficacy of home-made spacers is limited. This study was conducted to compare the efficacy of a valve-less home-made spacer with a commercial spacer in delivering salbutamol via MDI in acute asthma. Asthmatic children aged 5-15 years who presented with an acute exacerbation to the pediatric chest clinic of a tertiary care hospital were enrolled in a single blinded randomized parallel group study. The study patients received 10 puffs of salbutamol (100 microg/puff) via MDI-home-made spacer or MDI-commercial spacer. Pre and post inhalation measurements of peak expiratory flow rate (PEFR), oxygen saturation (SaO2), respiratory rate (RR), pulse rate (PR) were made and compared. Sixty children were enrolled in the study, 31 were administered salbutamol via the home-made spacer and 29 via the commercial spacer. The median increase in PEFR was similar in both the groups (20.8% vs 22.2%, p=0.4), clinical improvement being satisfactory in all patients. The valve-less home-made spacer is equally efficacious and cheaper than the commercial spacer in administering bronchodilators in acute exacerbations of asthma. Further studies on the efficacy of home-made spacer in delivery of inhaled steroids are needed.

Structural determinants of antiproliferative activity of heparin on pulmonary artery smooth muscle cells.

In addition to its anticoagulant properties, heparin (HP), a complex polysaccharide covalently linked to a protein core, inhibits proliferation of several cell types including pulmonary artery smooth muscle cells (PASMCs). Commercial lots of HP exhibit varying degrees of antiproliferative activity on PASMCs that may due to structural differences in the lots. Fractionation of a potent antiproliferative HP preparation into high and low molecular weight components does not alter the antiproliferative effect on PASMCs, suggesting that the size of HP is not the major determinant of this biological activity. The protein core of HP obtained by cleaving the carbohydrate-protein linkage has no growth inhibition on PASMCs, demonstrating that the antiproliferative activity resides in the glycosaminoglycan component. Basic sugar residues of glucosamine can be replaced with another basic sugar, i.e., galactosamine, without affecting growth inhibition of PASMCs. N-sulfonate groups on these sugar residues of HP are not essential for growth inhibition. However, O-sulfonate groups on both sugar residues are essential for the antiproliferative activity on PASMCs. In whole HP, in contrast to an earlier finding based on a synthetic pentasaccharide of HP, 3-O-sulfonation is not critical for the antiproliferative activity against PASMCs. The amounts and distribution of sulfonate groups on both sugar residues of the glycosaminoglycan chain are the major determinant of antiproliferative activity.

Effect of fully sulfated glycosaminoglycans on pulmonary artery smooth muscle cell proliferation.

Fully sulfated heparin and other glycosaminoglycans, namely heparan, chondroitin, and dermatan sulfates, and hyaluronan have been prepared by using sulfur trioxide under mild chemical conditions. All these derivatives were assayed for antiproliferative activity on cultured bovine pulmonary artery smooth muscle cells (BPASMCs). No appreciable difference was found between heparin and fully sulfated heparin. Chondroitin and dermatan sulfates actually stimulated BPASMCs growth but full sulfonation made them strongly antiproliferative. Native hyaluronan was not antiproliferative but became strongly so after sulfonation. Neither acharan sulfate nor N-sulfoacharan sulfate had any antiproliferative activity. This suggests that O-sulfonation of the polysaccharide is critical for antiproliferative activity, whereas N-sulfonation of glucosamine residues is not.

Bioactivity of marine organisms: Part VIII--Screening of some marine flora of western coast of India.

Methanolic extracts of 31 botanically identified species of marine flora, collected from Gujarat Coast, have been screened for a wide range of biological activities. Of these, 3 extracts showed anti-implantation, 2 had antiviral, 2 showed hypotensive, 1 had anti-inflammatory while 12 extracts showed diuretic activities. The antiviral activity; against EMCV, was confirmed in one alga. The active principles and results of these studies are reported.

Influence of molecular weight, protein core and charge of native heparin fractions on pulmonary artery smooth muscle cell proliferation.

Heparin macromolecules have been shown to inhibit cultured pulmonary artery smooth muscle cell proliferation in vitro and prevent hypoxic vascular remodeling in vivo. In an attempt to understand the structural determinants of heparin's antiproliferative properties, we have fractionated an antiproliferative preparation of commercial heparin into low and high molecular weight fractions. Then the high molecular weight heparin fraction was further fractionated on a DEAE-cellulose column by charge density eluting with 0 - 1 M NaCl linear gradient. The heparin protein peptides were both removed and isolated. These heparin fractions were assayed for antiproliferative effects on cultured bovine pulmonary artery smooth muscle cells. No appreciable differences were found among high and low molecular weight heparin fractions The core peptides showed no antiproliferative activity. However, higher charge density fraction was less antiproliferative.

Effects of oral lithium on the action of various C.N.S. active drugs.

Effects of prolonged lithium administration was seen on the action of various psychoactive drugs in animals. Apomorphine induced pecking in pigeons increased significantly by lithium treatment for 14 days, from 1445.3 +/- 202.5 in control to 2785.8 +/- 205.8 in Gp. B. Haloperidol-induced catalepsy score in albino rats increased significantly following chronic lithium treatment compared to control. Chlorpromazine-induced hypothermia in rabbits was immediate but transient, while in lithium treated rabbits induction of hypothermia was delayed, sustained and of greater magnitude. This action of lithium may be mediated by increasing the permeability of blood-brain barrier, or enhancing the sensitivity of alpha-adrenoceptors in brain.

Antiproliferative role of 3-O-sulfate glucosamine in heparin on cultured pulmonary artery smooth muscle cells.

Heparin macromolecules inhibit vascular remodeling associated with hypoxic pulmonary hypertension. Heparin's antiproliferative effect on smooth muscle cells, based on studies of synthetic pentasaccharide fragments, has been attributed to 3-O-sulfate on the internal glucosamine. To determine the role of 3-O-sulfation in smooth muscle cell growth, we treated three heparins of varying potency with heparitinases I and II, which degrade heparin fragments containing 3-O-sulfate on the glucosamine residue to delta-tetrasaccharides only. Our most antiproliferative heparin gave the least amount of delta-tetrasaccharides. This heparin was then fractionated according to degree of sulfation using ETOH precipitation. Again we found no antiproliferative difference between the highly sulfated fractions and those with a lesser degree of sulfation. These studies suggest that 3-O-sulfate of glucosamine residue is not critical in whole heparins for antiproliferative activity.

Inseparable iduronic acid-containing proteoglycan PG(IdoA) preparations of human skin and post-burn scar tissues: evidence for elevated levels of PG(IdoA)-I in hypertrophic scar by N-terminal sequencing.

Hypertrophic scarring is characterized by disordered collagen fibrils. In order to determine whether this is, in part, a result of changes in the population of proteoglycans that are thought to be involved in regulation of collagen fibril formation, we have compared PGs from post-burn normal and hypertrophic scar tissue, as well as from human dermis and epidermis. Efforts to separate the two major iduronic acid-containing proteoglycans, decorin [PG(IdoA)-II] and biglycan [PG(IdoA)-I], for quantitation were not successful. The different N-terminal sequences of these two iduronic acid-containing proteoglycans PG(IdoA-I and -II were utilized to estimate the relative amounts in the above PG(IdoA) preparations. Normal scar, dermis and epidermis were all found to contain primarily decorin with low (< 10%) levels of biglycan relative to decorin. In contrast, iduronic acid-containing proteoglycans from hypertrophic scar were found to be approximately 30% biglycan [PG(IdoA)-I]. This may be a proximal cause of altered collagen fibrils, or may result in alterations in the sequestration of growth factors, which then results in changes in collagen that effect the appearance of the scar. 1966 Elsevier Science Ltd.

Scrokoelziside A, a triterpene glycoside from Scrophularia koelzii.

Scrokoelziside A, isolated from the aerial parts of Scrophularia koelzii was shown to be 3-O-([alpha-L-rhamnopyranosyl-(1-->3), beta-D-glucopyranosyl-(1-->2)]-beta-D-fucopyranosyl-(1-->4)-beta-D- glucopyranosyl]-13 beta,28-epoxyolean-11-en-23-ol, on the basis one- and two-dimensional NMR homo- and hetero-nuclear spectroscopic evidence.

Differential effect of three commercial heparins on Na+/H+ exchange and growth of PASMC.

Heparin preparations vary in chemical content and in antiproliferative activity for pulmonary artery smooth muscle cells (PASMC). Intracellular alkalinization via stimulation of the Na+/H+ antiporter appears to be a permissive event for proliferation of PASMC. We wondered whether the variable effect of heparin preparations on PASMC growth might be due to different degrees of inhibition of the Na+/H+ antiporter and whether variations in chemical formulation might correlate with the inhibition. Fluorescent microscopy of bovine PASMC was done using a dye with which fluorescence varies directly with intracellular pH (pHi). Bovine PASMC were preincubated with three heparin preparations previously shown to vary in antiproliferative activity, at 1.0 microgram/ml for 24 h. Platelet-derived growth factor (PDGF; 60 ng/ml) on PASMC without heparin resulted in a rise in pHi of 0.27 +/- 0.02 pH units. The rise in pH units in heparin-treated PASMC was 0.34 +/- 0.03 with Choay, 0.21 +/- 0.02 with Elkins-Sinn, and 0.07 +/- 0.02 with Upjohn (+/-SE; all P < 0.05; n = 5). Upjohn heparin incubation for as little as 15 min still impeded the rise in pH induced by PDGF. Heparin did not block the Na+/H+ exchanger directly, as it still restored pHi in response to an acid load. Compared with PASMC proliferation induced by 60 ng/ml PDGF, 1 microgram/ml of Choay, Elkins-Sinn, and Upjohn heparin produced -4 +/- 7.4, 1.4 +/- 4.8, and 48 +/- 2.2% inhibition of PDGF control, respectively (P < 0.05 for Upjohn compared with PDGF and Choay). The heparins varied in protein content and amino acid composition. However, amino acid and glucosamine composition, total sulfation, and extent of 3-O-sulfation did not predict their activity. Thus inhibition of PDGF activation of the Na+/H+ antiporter by a given heparin preparation correlated well with its ability to inhibit PASMC proliferation.