Double J stent removal in paediatric patients by Vellore Catheter Snare technique: a randomised control trial.

BACKGROUND: Double J (DJ) stents placed at the end of paediatric urological procedures require another cystoscopy under general anaesthesia for removal. The second author developed a reproducible technique for snaring the DJ stent using an infant feeding tube (6-Fr or 8-Fr) and a 3-0 polypropylene suture per urethra. Having demonstrated the proof of concept, ethical clearance was obtained for an institutional randomised controlled trial. OBJECTIVE: The aim of the study was (1) to describe the Vellore Catheter Snare (VeCS) technique for DJ stent removal, (2) to study the efficacy of the technique and (3) to compare the costs of VeCS technique with cystoscopy on an intention-to-treat basis. STUDY DESIGN: The study design was that of a randomised control trial with parallel groups as a non-inferiority study. RESULTS: Forty children with unilateral indwelling DJ stents were enrolled from January to August 2018. They were randomised by unequal allocation (1:3) to cystoscopic and VeCS technique removal arms. The VeCS technique and cystoscopy were successful in 86.67% (26/30) and in 100% (10/10) cases, respectively, with no statistically significant difference in the outcome (p = 0.223). The average cost for cystoscopic removal of the stent was INR 14,579 and was INR 5636.5 for the VeCS technique (on an intention-to-treat basis). DISCUSSION: While per-urethral catheterisation is an outpatient/ward procedure in children, cystoscopy is not. Other techniques such as extraction strings and magnetic stents with their extraction device were found to have certain disadvantages. The VeCS technique, using common disposables, circumvented the need for inpatient admission, disinfected equipment usage and operation theatre time in 87% children, thereby reducing the costs incurred by the patient. CONCLUSION: The VeCS technique for DJ stent removal is a practical low-cost safe alternative to cystoscopic removal of DJ stents in children. Although the technique has a high success rate, it still needs the backup option of cystoscopy under general anaesthesia.

Effects of dietary fiber on the ideal standardized ileal digestible threonine:lysine ratio for twenty-five to fifty kilogram growing gilts.

Four experiments were conducted to determine effects of fiber on the ideal Thr:Lys ratio for 25- to 50-kg gilts. In Exp. 1, the objective was to determine the requirement for standardized ileal digestible Lys for gilts from 25 to 50 kg BW. Seventy gilts (24.54 ± 3.28 kg BW) were used in a growth assay with 2 pigs per pen, 5 diets, and 7 replicate pens per diet. The 5 diets were based on corn and soybean meal and contained between 0.80 and 1.32% SID Lys. Results indicated that 1.09% SID Lys was needed to optimize ADG and G:F. In Exp. 2, the objective was to determine the standardized ileal digestibility of AA in corn, soybean meal, field peas, fish meal, and soybean hulls. Six ileal-cannulated gilts (26.5 ± 0.74 kg BW) were allotted to a 6 × 6 Latin square design with 6 diets and 6 periods. Values for standardized ileal digestibility of AA were calculated for all ingredients. In Exp. 3, the objective was to determine the effect of fiber on the ideal SID Thr:Lys ratio for gilts from 25 to 50 kg BW. A total of 192 gilts (26.29 ± 4.64 kg BW) were used in a growth assay with 2 pigs per pen and 8 replicate pens per treatment. Six low-fiber diets and 6 high-fiber diets were formulated using the same batches of ingredients as in Exp. 2. Within each level of fiber, diets with SID Thr:Lys ratios ranging from 45:100 to 90:100 were formulated using the SID values calculated in Exp. 2. In both types of diets, ADG and G:F linearly and quadratically ( < 0.05) increased as the Thr:Lys ratio increased. Regression analysis estimated the ideal SID Thr:Lys ratio at 0.66 and 0.63 for ADG and G:F, respectively, for pigs fed low-fiber diets and at 0.71 and 0.63, respectively, for pigs fed high-fiber diets. In Exp. 4, the objective was to determine the N balance in pigs fed low-fiber or high-fiber diets that were formulated to have SID Thr:Lys ratios of 45:100 or 60:100. The 4 diets were formulated using the same batches of ingredients as in Exp. 2, and the SID values determined in Exp. 2 were used in diet formulations. Thirty-six gilts (29.0 ± 0.74 kg BW) were individually housed in metabolism crates with 9 replicate pigs per diet. Retention of N (% of intake) was greater (P < 0.05) for pigs fed the low-fiber diets compared with pigs fed the high-fiber diets regardless of the Thr:Lys ratio. Results of these experiments indicate that increased fiber levels in diets fed to growing gilts increase the requirement for Thr and that diets with higher fiber levels should be formulated to a greater SID Thr:Lys ratio.

Plasmonic gratings with nano-protrusions made by glancing angle deposition for single-molecule super-resolution imaging.

Super-resolution imaging has been advantageous in studying biological and chemical systems, but the required equipment and platforms are expensive and unable to observe single-molecules at the high (µM) fluorophore concentrations required to study protein interaction and enzymatic activity. Here, a plasmonic platform was designed that utilized an inexpensively fabricated plasmonic grating in combination with a scalable glancing angle deposition (GLAD) technique using physical vapor deposition. The GLAD creates an abundance of plasmonic nano-protrusion probes that combine the surface plasmon resonance (SPR) from the periodic gratings with the localized SPR of these nano-protrusions. The resulting platform enables simultaneous imaging of a large area without point-by-point scanning or bulk averaging for the detection of single Cyanine-5 molecules in dye concentrations ranging from 50 pM to 10 µM using epifluorescence microscopy. Combining the near-field plasmonic nano-protrusion probes and super-resolution technique using localization microscopy, we demonstrate the ability to resolve grain sizes down to 65 nm. This plasmonic GLAD grating is a cost-effective super-resolution imaging substrate with potential applications in high-speed biomedical imaging over a wide range of fluorescent concentrations.

Bladder calculi in the augmented bladder: a follow-up study of 160 children and adolescents.

INTRODUCTION: Bladder augmentation (BA) has been used for various congenital and acquired conditions to create a low pressure, continent catheterizable reservoir. The prevalence of calculi within the BA have been reported to be from 3 to 52.5%. The present study reports the prevalence and risk factors of bladder calculi in patients with BA. MATERIAL AND METHODS: A retrospective review of 160 patients was performed from January 1997 through December 2012. The various risk factors for the formation of bladder calculi such as the nature of the anatomical defect, presence of preoperative urinary calculi, type of bowel augmentation, addition of a mitrofanoff and/or bladder neck procedure, prevalence of post-operative urinary tract infections (UTIs), need for mitrofanoff revision due to stenosis/difficulty catheterization, postoperative significant hydronephrosis and bladder calculi were recorded for analysis. The children underwent open removal or endoscopic cystolithotripsy. One hundred and eight males and 52 females (average age 6.3 years) were followed up for a median of 70.5 months. All patients performed daily bladder irrigation with tap or drinking water. RESULTS: Post-operative bladder calculi were noted in 14 (8.8%) of 160 patients following BA. Median time to stone formation was 37.5 months (11-120 months). Recurrent febrile UTIs were noted in 16 of the 160 patients following BA. The various risk factors and their outcomes are summarized in table. Eight patients underwent open cystolithotomy and four patients were treated by cystolithotripsy. Post-operative recurrent bladder calculi were noted in 2 patients. Multivariate analysis revealed that exstrophy/epispadias (OR 17.2) and recurrent UTI (OR 55.4) were independent risk factors for developing postoperative calculi in bladder augmentations. All other risk factors did not achieve statistical significance. DISCUSSION: There seemed to be no difference in the prevalence of calculi in the ileal or colonic augmentations. Mucus secreted by the bowel segment blocks catheters leading to incomplete drainage, stagnation and UTIs. Our protocol consists of daily bladder irrigation till the effluents are clear of mucus. This is probably the key to the low prevalence of postoperative calculi (8.8%) in our patients. CONCLUSION: Bladder exstrophy/epispadias and UTIs are independent statistically significant risk factors for the formation of bladder calculi in BA patients. Other risk factors such as preoperative calculi, bladder neck procedures and the use of mitrofanoff though not statistically significant, may contribute to the overall risk. The performance of daily bladder irrigation is an important part of our management of mucus.

Anatomy of the fetal membranes using optical coherence tomography: part 1.

INTRODUCTION: In vitro studies on the structure of human fetal membranes have involved light or electron microscopy with fixation, dehydration, and staining. Recently, optical coherence tomography (OCT), an imaging technology, has provided high-resolution cross-sectional images of living biological tissues, with a penetration of 2-3 mm. We evaluated the use of this technology to examine the histologic features of human fetal membranes immediately after delivery. METHODS: Samples of fetal membranes of ten patients undergoing cesarean deliveries (four uncomplicated pregnancies, four with preeclampsia, and two with chorioamnionitis) and eight patients undergoing vaginal deliveries (six uncomplicated pregnancies and two with chorioamnionitis) were collected immediately after delivery. Samples were stretched across customized disks, rinsed, and analyzed using a time-domain OCT imaging system. Following OCT scanning, the samples were placed in formalin for histologic study. The OCT images were compared to histologic images of common human fetal membrane features. RESULTS: We were able to delineate the layers of the fetal membranes using bench-top time-domain OCT. The system was able to image histologic features of the fetal membranes, such as microscopic chorionic pseudocysts, ghost villi, meconium stained membranes, and chorioamnionitis. The OCT images corresponded with the histologic findings. DISCUSSION: This feasibility study demonstrates the potential of OCT technology for real-time assessment of human fetal membranes and may provide clinically useful information at delivery.

Nutritional value of dried fermentation biomass, hydrolyzed porcine intestinal mucosa products, and fish meal fed to weanling pigs.

Dried fermentation biomass (DFB) and hydrolyzed porcine intestinal mucosa are co-products of L-Lys • HCl production and heparin extraction, respectively. Three experiments were conducted to determine standardized ileal digestibility (SID) of AA (Exp. 1), concentration of DE and ME (Exp. 2), and standardized total tract digestibility (STTD) of P (Exp. 3) in DFB and 2 hydrolyzed porcine intestinal mucosa products (PEP50 and PEP2+), and compare these values with values for fish meal. In Exp. 1, 12 ileal cannulated barrows (BW = 11.5 ± 1.1 kg) were allotted to a replicated 6 × 6 Latin square design with 6 diets and 6 periods. A N-free diet, diet based on soybean meal (SBM), and 4 diets based on a combination of SBM and DFB, PEP50, PEP2+, or fish meal were formulated. With the exception of Lys, there were no differences in SID of indispensable AA between DFB and fish meal. Except for Thr, no differences in SID of indispensable AA between PEP50 and fish meal were observed, but SID of all indispensable AA, except Lys and Trp, was less (P < 0.05) in PEP2+ than in the other ingredients. In Exp. 2, 40 barrows (BW = 12.8 ± 1.4 kg) were allotted to 5 diets with 8 pigs/diet. A basal diet containing 96.4% corn and 4 diets containing corn and DFB, PEP50, PEP2+, or fish meal were formulated. The DE (5,445 kcal/kg DM) and ME (5,236 kcal/kg DM) in DFB were greater (P < 0.01) than in PEP50 (4,758 and 4,512 kcal/kg DM for DE and ME, respectively) and fish meal (4,227 and 3,960 kcal/kg DM for DE and ME, respectively). Also, DE in DFB was greater (P < 0.01) than in PEP2+ (4,935 kcal/kg DM), but ME in DFB was not different from that in PEP2+ (4,617 kcal/kg DM). Furthermore, DE in PEP50 and PEP2+ were greater (P < 0.01) than in fish meal, but ME did not differ from that in fish meal. In Exp. 3, 40 barrows (BW = 12.4 ± 1.3 kg) were randomly allotted to 5 diets with 8 pigs/diet. A P-free diet and 4 diets in which the sole source of P was from DFB, PEP50, PEP2+, or fish meal were formulated. The STTD of P in DFB (96.9%) and PEP2+ (97.6%) were greater (P < 0.01) than in PEP50 and fish meal (76.2% and 68.5%, respectively), and STTD of P in PEP50 was greater (P < 0.01) than in fish meal. In summary, SID of most indispensable AA did not differ among DFB, PEP50, and fish meal, but DE and ME and STTD of P in DFB were greater than in PEP50 and fish meal.

Reverse foam sclerotherapy of the great saphenous vein with sapheno-femoral ligation compared to standard and invagination stripping: a prospective clinical series.

OBJECTIVES: Comparison of Reverse Foam Sclerotherapy of the great saphenous vein (GSV) combed with sapheno-femoral junction (SFJ) ligation to standard (Babcock) stripping and invagination (Pin) stripping in a prospective clinical series. DESIGN: Prospective clinical series. MATERIALS AND METHODS: 90 consecutive limbs of 82 patients with incompetence of the GSV resulting in varicose veins were prospectively randomised into 3 groups of 30, treated by SFJ ligation and either reverse foam sclerotherapy, standard stripping or invagination stripping of the GSV. Outcomes were assessed post-operatively and at 2-weeks follow-up. Peri-operative blood loss (24 hrs), analgesic requirement, bruising and residual varicosities were assessed. Bruising was assessed by both patients and independent assessors using questionnaires. RESULTS: SFJ ligation plus reverse foam sclerotherapy of the GSV was associated with significantly less blood loss, bruising and post-op discomfort than either of the stripping techniques. (p<0.001, Mann-Whitney) CONCLUSION: Standard stripping of the GSV and invagination stripping are not associated with major discomfort and problems in the early post-operative period. SFJ ligation and GSV reverse foam sclerotherapy yielded greater patient satisfaction with less post-op bruising and discomfort and reduced analgesic requirements.

Blocked D phenomenon, a rare condition with Rh D haemolytic disease of newborn - a case report.

Accurate Rh testing can be difficult if the red cells are heavily coated with IgG anti D antibodies - a phenomenon called blocked D. Repeatedly, Rh D negative blood group report was obtained in a newborn male baby with severe haemolytic disease and features of kernicterus born to a 2nd gravida B Rh D negative mother. On investigation, the baby was grouped as B Rh D negative by direct grouping, but after elution, D antigen was detected and phenotyped as CcDe. Antibody was identified as anti D. All D antigens of the baby were fully saturated with anti D leaving any antigen to bind with antisera. Direct Coombs test was strongly positive even after three exchange transfusions. The baby also had free antibody apart from the red cell bound and the red cell eluate, gave a titre of 512. The mother was grouped as B Rh D negative and phenotyped as ce. She had IgM and IgG class of anti D with titres 32 and 1024 respectively. She also had IgM anti C (only in neat) and IgG anti-A with a titre of 512.

Does the establishment of a Vascular Unit reduce the prevalence of ruptured aortic aneurysms?

AIM: The aim of this study was to assess the effect of a specialised Vascular Unit upon the prevalence of ruptured aortic aneurysms in the same population catchments' area and associated mortality rates. SETTING: prospective computerised data collection from 1997 to today, retrospective from 1990-1996. SUBJECTS: 108 aneurysms were operated upon from 1990-1996, compared to 317 from 1997 to Aug 2002. MAIN OUTCOME MEASURES: statistical analysis was done using SPSS statistics with Kaplan Meier life table curves and compared by the log rank test while the Mann Whitney test was used for comparison of mortality. RESULTS: The median values for ruptured aneurysms per year were 5 for the early period, compared to 10 for the recent years, while the median values for both urgent and ruptured were 7 and 18.5 cases annually, respectively. The number of scheduled procedures increased by 500% in the second period, with median values of 7 and 36.5, respectively. In-hospital mortality according to category was 21% for scheduled, 31% for urgent and 69% for ruptured aneurysms in the early period, compared to 3.7%, 16% and 29% respectively, following the establishment of the Vascular Unit. CONCLUSION: Despite the five-fold increase in the total number of aortic aneurysm repairs (as expected), the number of ruptured aneurysms operated upon increased as well. There was just a trend for a reduction in the absolute numbers of ruptured aneurysms operated upon in the last 2 years. Mortality, on the other hand, decreased dramatically in all categories, with the overall 30-day mortality decreasing more than four-fold, from 40% to 9.3%, while the respective mortalities according to the category of intervention were 3.7% vs 21% for scheduled, 16% vs 35% for urgent and 29% vs 69% for ruptured aneurysms, with a P value of less than 0.01. However, there was no difference in the numbers of patients with ruptured aneurysm reaching the hospital (operated or not) between the two periods (median values of 11 and 10.5 annually). The presence of a Vascular Unit, although it achieves dramatically better results, is not associated with a reduction in the number of emergency proceduresaeat least in the intermediate termaedespite an expansion in the indications for surgery, increased awareness and prompt referrals (centralisation).

Normal human plasma anti-beta-glucoside antibody has markedly elevated IgA content and binds fungal and yeast polysaccharides.

Normal human plasma antibody that recognizes beta-linked glucoside moiety was purified by affinity chromatography on cellulose. The anti-beta-glucoside antibody had three times higher IgA to IgG ratio and substantially higher polymeric IgA content than total serum immunoglobulins. Cellobiose and other beta-glucosides were best inhibitors of its binding to polystyrene microwell-coated polysaccharides. In synthetic glycoproteins made by conjugating disaccharides to hemoglobin or bovine serum albumin, cellobiose, unlike lactose or maltose, was sugar-specifically recognized by the antibody. It also recognized polystyrene well-coated beta1-->3 linked glycans of Saccharomyces cerevisiae, Candida albicans and of barley in decreasing order of affinity. Its sugar-binding site could thus accommodate beta-glucoside with or without substitution at C4 and C3. High IgA content along with the capacity to bind common microbial and dietary antigens pointed to the immune inflammatory potential of the antibody.

Y-duplication of the male urethra: use of anterior anorectal wall for posterior urethral lengthening.

We have approached two patients with Y-duplication of the male urethra by a new two-staged technique to provide better results. A strip of anterior anorectal wall in continuity with the posterior urethra was used for posterior urethral lengthening and a tubed pedicled prepucial flap was used to reconstruct the anterior urethra without using the native urethra. This was done under a covering colostomy. After a gap of 6 months to allow for healing of the anorectum and to ensure adequate functioning of the perineal neourethra, second stage reconstruction was done using buried scrotal tube for the mid urethra along with colostomy closure. On follow-up at 8 and 12 months, respectively, both children were well with no stricture or fistula. There was normal anal continence and no stenosis. This technique tackles the problem in Y-duplication of the male urethra of lengthening the posterior urethral channel, which is often difficult to bring to the anterior half of the perineum especially if the opening is high up in the anorectum (case 2).

Molecular mechanisms of water and solute transport across archaebacterial lipid membranes.

Archaebacteria thrive in environments characterized by anaeobiosis, saturated salt, and both high and low extremes of temperature and pH. The bulk of their membrane lipids are polar, characterized by the archaeal structural features typified by ether linkage of the glycerol backbone to isoprenoid chains of constant length, often fully saturated, and with sn-2,3 stereochemistry opposite that of glycerolipids of Bacteria and Eukarya. Also unique to these bacteria are macrocyclic archaeol and membrane spanning caldarchaeol lipids that are found in some extreme thermophiles and methanogens. To define the barrier function of archaebacterial membranes and to examine the effects of these unique structural features on permeabilities, we investigated the water, solute (urea and glycerol), proton, and ammonia permeability of liposomes formed by these lipids. Both the macrocyclic archaeol and caldarchaeol lipids reduced the water, ammonia, urea, and glycerol permeability of liposomes significantly (6-120-fold) compared with diphytanylphosphatidylcholine liposomes. The presence of the ether bond and phytanyl chains did not significantly affect these permeabilities. However, the apparent proton permeability was reduced 3-fold by the presence of an ether bond. The presence of macrocyclic archaeol and caldarchaeol structures further reduced apparent proton permeabilities by 10-17-fold. These results indicate that the limiting mobility of the midplane hydrocarbon region of the membranes formed by macrocyclic archaeol and caldarchaeol lipids play a significant role in reducing the permeability properties of the lipid membrane. In addition, it appears that substituting ether for ester bonds presents an additional barrier to proton flux.

Children with persistent conduct problems who dropout of treatment.

Dropout of treatment is one of the key issues in outcome in a child and adolescent mental health service. We report two studies focusing on the treatment process and the dropout rate of children with persistent conduct problems presenting to a community mental health service, using a prospective design. The first study included 32 children and used a randomised controlled treatment design comparing a CBT approach with conjoint family therapy and an eclectic approach. The overall dropout rate was 36%. Dropout occurred significantly less frequently in the CBT group. The dropout group was associated with mothers who were younger and less educated, a poorer rating by the clinicians at the last meeting, parental dissatisfaction with the treatment service and perception that the treatment was less organised and having less behavioural tasks. In the second study we used a naturalistic follow-up design. Forty-six children were included. The overall dropout rate was 48%. Again, the children who defaulted were rated by clinicians as less likely to have improved and dropout was also significantly associated with parental perception of a less organised treatment. In both studies dropout usually occurred after assessment and at the early phase of treatment.

Glutaraldehyde cross-linking of lectins to marker enzymes: protection of binding site by specific sugars.

The role of bound specific sugars in protecting the sugar binding activity of several galactose binding proteins during their covalent conjugation to horse radish peroxidase by glutaraldehyde-mediated cross-linking was examined by: a) affinity matrix binding of the conjugate, b) enzyme linked lectin assay and c) hemagglutination assay. During conjugation using 1% glutaraldehyde, protection of jack fruit (Artocarpus integrifolia) lectin (jacalin) activity depended on concentration of specific sugar present during conjugation; optimum protection was offered by 50 mM galactose. This indicated the presence of one or more primary groups at the binding site of jacalin, which is (are) essential for sugar binding. On the other hand, such essential amino group(s) was not indicated at the sugar binding site of the peanut lectin, bovine heart galectin or of the human serum anti alpha-galactoside antibody, since exclusion of sugar during their conjugation to HRP did not diminish sugar binding activity. The differential behavior is discussed in the light of reported differences in sugar specificities. Results indicated that sugar mediated blocking of active site may be used in characterization of the latter in lectins.

Hepatotoxicity due to troglitazone: report of two cases and review of adverse events reported to the United States Food and Drug Administration.

Two patients (a 48-year-old woman and a 62-year-old man) developed clinical and laboratory signs of hepatotoxicity due to troglitazone (Rezulin), a thiazolidinedione used in treatment of diabetes mellitus. There was no clear clinical evidence of drug allergy, although the woman experienced colitis before the onset of recognized hepatotoxicity. Liver biopsies showed bridging necrosis and fibrosis in the woman and hepatitis with granuloma formation in the man. The abnormalities in liver chemistries resolved promptly upon cessation of the drug. Cases involving 46 patients reported to the United States Food and Drug Administration are also reviewed. Troglitazone is a useful new oral antihyperglycemic agent, but in about 1.9% of patients hepatotoxicity has occurred, which may be severe and even fatal. Frequent monitoring of serum liver chemistries in patients taking the drug is essential.

Rosai dorfman disease - an unusual presentation: A case report.

Rosai Dorfman Disease (Sinus Histiocytosis with massive lymphadenopathy) is a rare self-limiting histiocytic disorder of unknown etiology usually presenting with cervical lymphadenopathy and haematological abnormalities. Extra-nodal presentation is rare . This ease is such a rare presentation of Rosai Dorfman Disease.

IFN-alpha receptor mRNA expression in a United States sample with predominantly genotype 1a/I chronic hepatitis C liver biopsies correlates with response to IFN therapy.

Our aim was to assess whether, in the United States, with the predominant hepatitis C viral (HCV) genotypes 1a/I and 1b/II, hepatic interferon-alpha receptor (IFNAR) mRNA expression correlated with response to IFN therapy, levels of HCV RNA, or histologic activity index (HAI). Nine of 24 patients (38%) had an initial response to IFN treatment, 5 of whom (21%) had a sustained response. The corrected hepatic IFNAR mRNA expression (measured by RT-PCR) for the sustained responder group (mean +/- SE, 0.16 +/- 0.06, n = 5) was significantly higher than for the nonresponding group (0.059 +/- 0.01, n = 15) (p < 0.02). Patients who relapsed had an intermediate value (0.092 +/- 0.029, n = 4). Higher IFNAR expression was inversely correlated with a lower serum HCV RNA titer (p < 0.01), and responders to IFN treatment tended to have a lower titer of HCV RNA (p = 0.056). We found no significant correlation between the amounts of IFNAR with (1) the total HAI (low HAI < or = 7, IFNAR 0.076 +/- 0.013, n = 10; high HAI > or = 8, IFNAR 0.092 +/- 0.027, n = 14, ns) or (2) individual inflammation, necrosis, or fibrosis components of the HAI. As with Japanese HCV patients with genotypes 1b/II-2b/IV, higher hepatic IFNAR mRNA expression in the United States with predominant genotypes 1a/I and 1b/II appears to correlate with response to IFN therapy and a low HCV RNA titer but not with the total HAI or its components.

Water transport across yeast vacuolar and plasma membrane-targeted secretory vesicles occurs by passive diffusion.

To determine whether solute transport across yeast membranes was facilitated, we measured the water and solute permeations of vacuole-derived and late secretory vesicles in Saccharomyces cerevisiae; all permeations were consistent with passive diffusive flow. We also overexpressed Fps1p, the putative glycerol facilitator in S. cerevisiae, in secretory vesicles but observed no effect on water, glycerol, formamide, or urea permeations. However, spheroplasts prepared from the strain overexpressing Fps1p showed enhanced glycerol uptake, suggesting that Fps1p becomes active only upon insertion in the plasma membrane.

CXCR4 mRNA expression in colon, esophageal and gastric cancers and hepatitis C infected liver.

We have recently demonstrated by Northern blot and RT-PCR that the mRNA expression of the alpha-chemokine hIRH/SDF-1alpha is reduced in hepatocellular carcinoma (HCC), several digestive tract cancers and premalignant colon adenomas, and that its receptor CXCR4 mRNA expression is reduced in HCC. Here we investigate the expression of CXCR4 mRNA expression in several digestive tract cancers and hepatitis C viral (HCV) infected liver, a premalignant condition. There was no difference in the CXCR4 mRNA expression in colon, esophageal or gastric cancers compared to non-cancerous tissues. This is significantly different from the reduced expression we have seen with hepatocellular carcinoma (p<0.05). To better refine regional tumor or hepatic cytokine mRNA analysis within a biopsy sample we describe a micro-isolation technique for RNA extraction from portal and triad areas of liver biopsies or other small malignant or non-malignant biopsy samples suitable for use in RT-PCR and differential display reactions. In HCV liver biopsies, the expression of hIRH and its receptor CXCR4 mRNA, corrected for G3PDH, was not significantly different from that of control non-HCV (steatosis) biopsies. CXCR4 is expressed on leukocytes and its expression was predicted to correlate with hepatic inflammation. CXCR4 receptor mRNA expression did correlate significantly with that of its ligand hIRH/SDF-1alpha (p=0.001), and with the severity of fibrosis (p<0.05), but not with portal inflammation (p<0.10), piecemeal necrosis (p<0.10), lobular inflammation (p>0.10), the presence of lymphoid aggregates (p>0.10), or the total histological activity index (p=0.07). There was no difference in expression of hIRH or CXCR4 between responders and non-responders to interferon (IFN) treatment, while as a control, the responder group of patients did show a higher expression of IFNalpha receptor than the non-responder group (p=0.05).