Neourethra creation using autologous vascularized intestinal graft in cats.

The objective of this study was to assess the application of a urethrostomy technique that utilizes an autologous vascularized intestinal graft as a "neourethra" and to analyze its short- and long-term feasibilities. Six cats with urethral rupture and 8 cats with urethral stricture and a history of urethrostomy were included. The inclusion criteria were the indication for urethroplasty and limited urethral length for perineal urethrostomy. A segment of intestine was prepared as a graft for urethral repair. The diameter of the aboral end was adjusted to facilitate anastomosis with the urethra or neck of the urinary bladder. An ostomy was created in the prepubic region using the oral end. The postoperative follow-up period was at least 1 y. Restoration of urinary flow was successful in all cases immediately after surgery. During postoperative follow-up, minimal complications were observed, with urinary incontinence being one of the most frequent, occurring in 28.5% (4/14) of the cases. Urine culture, carried out at different times during the follow-up period, was positive in 72.7% (8/11) of the cats. The autologous graft from the vascularized intestinal segment was an appropriate urethral substitute and this urethroplasty technique therefore proved to be feasible in cats. The postoperative complications observed were not exclusive to this technique and generally could either be corrected or tolerated. Periodic clinical follow-up examinations are recommended. This procedure allows the reestablishment of urinary flow and should be considered a favorable option, particularly when there is insufficient urethral tissue to allow repair using conventional techniques.

L'objectif de cette étude était d'évaluer l'application d'une technique d'urétrostomie qui utilise une greffe intestinale vascularisée autologue comme « néo-urètre ¼ et d'analyser ses faisabilités à court et à long terme. Six chats avec rupture urétrale et huit chats avec rétrécissement urétral et antécédents d'urétrostomie ont été inclus. Les critères d'inclusion étaient l'indication d'urétroplastie et la longueur urétrale limitée pour l'urétrostomie périnéale. Un segment d'intestin a été préparé comme greffon pour la réparation urétrale. Le diamètre de l'extrémité aborale était ajusté pour faciliter l'anastomose avec l'urètre ou le col de la vessie. Une stomie a été créée dans la région prépubienne en utilisant l'extrémité orale. La période de suivi postopératoire était d'au moins 1 an. La restauration du flux urinaire a réussi dans tous les cas immédiatement après la chirurgie. Au cours du suivi postopératoire, des complications minimes ont été observées, l'incontinence urinaire étant l'une des plus fréquentes, survenant dans 28,5 % (4/14) des cas. La culture urinaire, réalisée à différents moments de la période de suivi, était positive chez 72,7 % (8/11) des chats. La greffe autologue du segment intestinal vascularisé était un substitut urétral approprié et cette technique d'urétroplastie s'est donc avérée réalisable chez le chat. Les complications postopératoires observées n'étaient pas exclusives à cette technique et pouvaient généralement être soit corrigées soit tolérées. Des examens de suivi clinique périodiques sont recommandés. Cette procédure permet le rétablissement du flux urinaire et doit être considérée comme une option favorable, en particulier lorsque le tissu urétral est insuffisant pour permettre une réparation à l'aide de techniques conventionnelles.(Traduit par Docteur Serge Messier).

Effect of anticoagulant adjustment on prothrombin time test using two different PT reagents in patients with elevated hematocrit.

The recommendations for adjustment of citrate volume in sample tubes with high hematocrit (Ht) are based on indirect studies of underfilled tubes or artificially constructed Ht values. The aim of this study was to evaluate the effect of citrate volume adjustment in sample tubes from patients with hematocrit >55% using two different prothrombin time (PT) tests. METHODS: Paired citrate-adjusted and unadjusted blood specimens were obtained from 181 patients from the pulmonary hypertension ambulatory with high Ht values and on warfarin therapy. The samples were tested using recombinant human tissue factor (RTF) and reagents extracted from rabbit brain (HS Plus). The results are expressed as the international normalized ratio (INR). The correlation and percent change (% change) between sample pairs were calculated. RESULTS: INR-RTF results from adjusted and unadjusted citrate blood specimens showed a strong correlation (R2 â€‹= â€‹0.8226, p â€‹< â€‹0.0001). The INR median was 2.25 (95% CI 2.10 to 2.41) for citrate-adjusted samples and was 2.22 (95% CI 2.06 to 2.38) for citrate-unadjusted samples. For samples with Ht >62%, the % change between sample pairs was >10%. Results using HS Plus showed a moderate correlation between citrate-adjusted and unadjusted samples (R2 â€‹= â€‹0.4267, p â€‹< â€‹0.0001). The INR median was 2.51 (95% CI 2.35 to 2.68) for citrate-adjusted samples and 3.45 (95% CI 3.11 to 3.80) for citrate-unadjusted samples. For samples with Ht>55%, the % change between sample pairs was higher than 10%. CONCLUSION: Our data demonstrate that in patients with polycythemia on warfarin therapy, INR-RTF does not require anticoagulant adjustment for assessment of samples with Ht <62%.

Routine Coagulation Tests in Patients With Nonvalvular Atrial Fibrillation Under Dabigatran and Rivaroxaban Therapy: An Affordable and Reliable Strategy?

Dabigatran and rivaroxaban, direct oral anticoagulants (DOACs), affect coagulation tests, and knowledge of their effects is important for therapeutic monitoring. Our aim was to examine the association between DOAC levels and routine coagulation tests in patients with nonvalvular atrial fibrillation. Samples from patients receiving dabigatran (150 mg) and patients receiving rivaroxaban (20 mg) were collected 2 hours after drug intake. Direct oral anticoagulant concentrations were determined using direct Hemoclot thrombin inhibitor (HTI) assay (HTI test) and a direct Xa inhibitor (Anti Xa-Riva). The routine coagulation measured included activated partial thromboplastin time (aPTT) and prothrombin time (PT). The median plasmatic dabigatran was 128.3 ng/mL (95% confidence interval [CI]: 93.7-222.6 ng/mL). The HTI exhibited a good correlation with aPTT ( R2 = 0.74; P < .0001). The median plasmatic rivaroxaban was 223.9 ng/mL (95% CI: 212.3-238.9 ng/mL). Anti-Xa-Riva correlated with PT ( R2 = 0.69, P< .0001) and aPTT (R2 = 0.36, P < .001), but prolonged PT results were obtained, even below the rivaroxaban therapeutic range (20%). The routine coagulation tests were able to identify out of therapeutic range concentrations for dabigatran and rivaroxaban. We suggest the use of these screening tests to better understand and monitor the subtherapeutic concentrations of these DOACs.

Treatment of Bisphenol A (BPA) in water using UV/H2O2 and reverse osmosis (RO) membranes: assessment of estrogenic activity and membrane adsorption.

Removal of an endocrine disrupting compound, Bisphenol A (BPA), from water was investigated using two treatment processes, UV/H2O2 advanced oxidation (AOP) and reverse osmosis (membrane separation). Furthermore, changes in estrogenic activity using in vitro yeast estrogen screen assay as well as the adsorption of BPA by the membrane surface were evaluated. The best UV/H2O2 performance was obtained using the highest established values of all parameters, reaching 48% BPA removal. Within the investigated conditions of the AOP, when lower doses of UV were used, a higher removal efficiency was achieved at a higher initial concentration of BPA. However, the same behavior was not observed for the highest UV dose, in which the removal efficiency was not dependent on BPA initial concentration. In both cases, removal efficiency increased as H2O2 concentration increased. The formation of estrogenic by-products was observed in UV/H2O2. The membrane rejection efficiency varied from 60% to 84% and all experiments showed adsorption of BPA by the membrane surface. The RO membrane showed a greater BPA removal efficiency for samples containing 10 µg·L-1 than UV/H2O2 at the evaluated treatment conditions.

Chitosan coated nanostructured lipid carriers (NLCs) for loading Vitamin D: A physical stability study.

A nanostructured lipid carrier (NLC) has been developed as a loading system for Vitamin D (VD). The NLCs were obtained by melt-emulsification method and coated with chitosan (CHI) by electrostatic deposition. The lipids used in the formulations were selected in order to provide higher encapsulation efficiency. Thermophysical properties of particles were evaluated by differential scanning calorimetry; particle stability was characterized by size distribution, polydispersity index, zeta-potential and light backscattering. The coating over NLCs was carried out by potentiometric titration with CHI at concentrations of 1.0, 1.5, 2.0 and 2.5% (w/v). Stearic (SA) and oleic acids (OA) were the lipids that showed higher compatibility with VD. The NLC 70(SA):30(OA) was the particle with the lowest polydispersity, size variation and less tendency to physical instability during the storage time. This formulation also presented encapsulation efficiency higher than 98%. In the particles coating, CHI adsorption into the colloidal dispersion provided an initial electrostatic stabilization of the system. A long-term steric barrier was established through further incorporation of CHI. Coated NLCs showed a core-shell structure and a positive zeta-potential (+30 mV), remaining stable for 60 days at 25 °C. During storage time, no expulsion of VD out of the particle was observed.

Obstructive sleep apnea and effects of continuous positive airway pressure on triglyceride-rich lipoprotein metabolism.

This study aimed to explore lipoprotein metabolism in obstructive sleep apnea (OSA) and the effects of continuous positive airway pressure (CPAP). We studied 15 men with severe OSA [apnea-hypopnea index (AHI) ≥30 events/hour] and 12 age-, BMI-, and waist circumference-matched volunteers without OSA (AHI <5 events/hour). Carotid intima-media thickness (CIMT) was determined by a blind examiner. After 12 h fasting, a triglyceride-rich chylomicron-like emulsion, labeled with [14C]cholesteryl oleate and [3H]triolein, was injected intravenously followed by blood sample collection at preestablished times. Fractional clearance rate (FCR) of the radiolabeled lipids was estimated by compartmental analysis of radioisotope decay curves. Compared with controls, patients with OSA showed a significant delay in both cholesteryl ester FCR (0.0126 ± 0.0187 vs. 0.0015 ± 0.0025 min-1; P = 0.0313) and triglycerides FCR (0.0334 ± 0.0390 vs. 0.0051 ± 0.0074 min-1; P = 0.0001). CIMT was higher in the OSA group: 620 ± 17 vs. 725 ± 29 µm; P = 0.004. Cholesteryl ester FCRs were inversely related to total sleep time <90% (r = -0.463; P = 0.029) and CIMT (r = -0.601; P = 0.022). The triglyceride FCR was inversely correlated with AHI (r = -0.537; P = 0.04). In a subgroup of patients treated with CPAP for 3 months (n = 7), triglyceride FCR increased 5-fold (P = 0.025), but the cholesteryl ester FCR was unchanged. In conclusion, severe OSA decreased lipolysis of triglyceride-rich lipoproteins and delayed removal of remnants. CPAP treatment may be effective to restore the lipolysis rates.

Determination of anthocyanins and non-anthocyanin polyphenols by ultra performance liquid chromatography/electrospray ionization mass spectrometry (UPLC/ESI-MS) in jussara (Euterpe edulis) extracts.

This work aimed to propose two analytical methods for the quantitative and qualitative analysis of major anthocyanins and non-anthocyanin phenolic compounds in jussara (Euterpe edulis) extracts, using ultra performance liquid chromatography-mass spectrometry. These methods were evaluated for selectivity, precision, linearity, detection and quantification limits. The complete separation of 5 anthocyanins and 22 non-anthocyanins polyphenols was achieved in 4.5 and 7 min, respectively. Limits of detection ranged from 0.55 to 9.24 µg/L, with relative standard deviation for concentration up to 7.0%. In jussara extract, 13 of the 27 analytes were characterized. The dominant compound was cyanidin-3-O-rutinoside, representing about 73% of the total phenolic compounds content (approximately 23 mg/g of extract in dry weight). Other phenolic compounds found in the extract were: cyanidin-3-O-glucoside, pelargonidin-3-O-glucoside, quercetin, rutin, myricetin, kaempferol, kaempferol-3-O-rutinoside, luteolin, apigenin, catechin, ellagic acid and 4,5-dicaffeoylquinic acid.

Plasma lipids, lipoprotein metabolism and HDL lipid transfers are equally altered in metabolic syndrome and in type 2 diabetes.

Metabolic syndrome (MetS) refers to states of insulin resistance that predispose to development of cardiovascular disease and type 2 diabetes (T2DM). The aim was to investigate whether plasma lipids and lipid metabolism differ in MetS patients compared to those with T2DM with poor glycemic control (glycated hemoglobin > 7.0). Eighteen patients with T2DM, 18 with MetS and 14 controls, paired for age (40-70 years) and body mass index (BMI), were studied. Plasma lipids and the kinetics of a triacylglycerol-rich emulsion labeled with [(3)H]-triolein ([(3)H]-TAG) and [(14)C]-cholesteryl esters ([(14)C]-CE) injected intravenously followed by one-hour blood sampling were determined. Lipid transfers from an artificial nanoemulsion donor to high-density lipoprotien (HDL) were assayed in vitro. Low-density lipoprotein (LDL) and HDL cholesterol (mg/dl) were not different in T2DM (128 ± 7; 42 ± 7) and MetS (142 ± 6; 39 ± 3), but triacylglycerols were even higher in MetS (215 ± 13) than in T2DM (161 ±11, p < 0.05). Fractional clearance rate (FCR, in min(1)) of [(3)H]-TAG and [(14)C]-CE were equal in T2DM (0.008 ± 0.018; 0.005 ± 0.024) and MetS (0.010 ± 0.016; 0.006 ± 0.013), and both were reduced compared to controls. The transfer of non-esterified cholesterol, phospholipids and triacylglycerols to HDL was higher in MetS and T2DM than in controls (p < 0.01). Cholesteryl ester transfer and HDL size were equal in all groups. Results imply that MetS is equal to poorly controlled T2DM concerning the disturbances of plasma lipid metabolism examined here, and suggest that there are different thresholds for the insulin action on glucose and lipids. These findings highlight the magnitude of the lipid disturbances in MetS, and may have implications in the prevention of cardiovascular diseases.

Effect of exercise training on plasma levels and functional properties of high-density lipoprotein cholesterol in the metabolic syndrome.

Intense lifestyle modifications can change the high-density lipoprotein (HDL) cholesterol concentration. The aim of the present study was to analyze the early effects of short-term exercise training, without any specific diet, on the HDL cholesterol plasma levels and HDL functional characteristics in patients with the metabolic syndrome (MS). We studied 30 sedentary subjects, 20 with and 10 without the MS. The patients with the MS underwent moderate intensity exercise training for 3 months on bicycle ergometers. Blood was sampled before and after training for biochemical analysis, paraoxonase-1 activity, and HDL subfraction composition and antioxidative capacity. Lipid transfer to HDL was assayed in vitro using a labeled nanoemulsion as the lipid donor. At baseline, the MS group had greater triglyceride levels and a lower HDL cholesterol concentration and lower paraoxonase-1 activity than did the controls. Training decreased the plasma triglycerides but did not change the low-density lipoprotein or HDL cholesterol levels. Nonetheless, exercise training increased the HDL subfractions' antioxidative capacity and paraoxonase-1 activity. After training, the MS group had compositional changes in the smallest HDL subfractions associated with increased free cholesterol and cholesterol ester transfers to HDL, reaching normal values. In conclusion, the present investigation has added relevant information about the dissociation between the quantitative and qualitative aspects of HDL after short-term exercise training without any specific diet in those with the MS, highlighting the importance of evaluating the functional aspects of the lipoproteins, in addition to their plasma levels.

Optimization of the enzymatic hydrolysis of mussel meat.

UNLABELLED: Mussel meat was subjected to enzymatic hydrolysis using Protamex. The relationship of temperature (46 to 64 degrees C), enzyme : substrate ratio (0.48% to 5.52%), and pH (6.7 to 8.3) to the degree of hydrolysis were determined. The surface response methodology showed that the optimum conditions for enzymatic hydrolysis of mussel meat were pH 6.85, temperature 51 degrees C, and enzyme : substrate ratio of 4.5%. Under these conditions a degree of hydrolysis of 26.5% and protein recovery of 65% were obtained. The produced hydrolysate, under optimum condition, was characterized in terms of chemical composition, electrophoretic profile, and amino acid composition. PRACTICAL APPLICATION: The practical application of mussel meat hydrolysate is its use as flavoring in products such as soups, sauces, and special beverages. In addition, the product is partially digested and has great nutritional value due to its good amino acid profile and thus can be used as a food supplement in special diets.

Cholestasis induced by model organic anions protects from acetaminophen hepatotoxicity in male CD-1 mice.

Administration of the non-metabolizable organic anion indocyanine green (ICG) prior to a toxic dose of acetaminophen (4-acetamidophenol; APAP) reduces liver injury 24h after dosing. ICG also produces a dose-dependent decrease in bile flow in mice and rats. Studies in bile duct-cannulated rats suggest that cholestasis can play a role in this protection. This study was conducted to determine if the ability of model organic anions to produce cholestasis is relevant to the protection against APAP hepatotoxicity afforded by ICG. In these studies, overnight fasted male CD-1 mice were dosed (i.v.) with the cholestatic dyes bromcresol green (BCG, 30 micromol/kg) and rose bengal (RB, 60 micromol/kg) immediately prior APAP administration (500 mg/kg, i.p.). Other groups of mice received the non-cholestatic dyes dibromosulphthalein (DBSP, 150 micromol/kg) and amaranth (AM, 300 micromol/kg) prior to APAP. Controls were given vehicle only. Hepatocellular necrosis was evident at 24 h in control mice receiving APAP. Pretreatment with the cholestatic dyes BCG and RB decreased the severity of hepatocellular necrosis induced by APAP. However, administration of the non-cholestatic dyes DBSP and AM did not alter APAP-induced liver damage. Glutathione replenishment was not altered by pretreatment with any of these dyes. Furthermore, ICG protected mice against carbon tetrachloride (CCl4) hepatotoxicity. Since CCl4 undergoes minimal biliary excretion and does not compete for biliary transport function, this finding supports the notion that cholestasis itself rather than competition for canalicular transporters is central to the hepatoprotection by ICG and other cholephilic dyes.

Transport deficient (TR-) hyperbilirubinemic rats are resistant to acetaminophen hepatotoxicity.

The biliary excretion of acetaminophen (APAP) is reduced in transport deficient (TR-) hyperbilirubinemic rats lacking the multidrug resistance-associated protein 2 (Mrp2). This mutant strain of Wistar rats has impaired biliary excretion of organic anions and increased hepatic glutathione. The rational for this study was to determine if there is an altered risk for liver damage by APAP in the absence of Mrp2. Therefore, the susceptibility of TR- rats to APAP hepatotoxicity was investigated. Male Wistar and TR- rats were fasted overnight before APAP treatment (1 g/kg). Hepatotoxicity was assessed 24 h later by plasma sorbitol dehydrogenase activity and histopathology. In other studies, TR- rats received buthionine sulfoximine before APAP to reduce hepatic glutathione to values similar to those in Wistar rats. mRNA expression of APAP metabolizing enzymes was also measured in naïve animals. Wistar rats treated with APAP showed significant elevations in plasma sorbitol dehydrogenase activity, while no increases in enzyme activity were observed in TR- rats. Histopathology was in agreement. Hepatic non-protein sulfhydryls were significantly lower in Wistar rats receiving APAP than in TR- rats. TR- rats treated with buthionine sulfoximine and APAP showed dramatic increases in hepatotoxicity. TR- rats had increased mRNA expression of several APAP metabolizing enzymes. Mrp2 expression not only is important in biliary excretion, but also influences the toxic potential of reactive intermediates by controlling intrahepatic GSH and possibly drug metabolism.

The rat ear vein model for investigating in vivo thrombogenicity of ultrafine particles (UFP).

Recent studies in rodents indicate that intravenous or intratracheal administration of ultrafine particles (UFP) increases thrombogenesis in a surgically exposed peripheral vein after photodynamic excitation of intravenously injected rose bengal (RB). We sought to adapt the invasive peripheral vein RB model to a noninvasive monitoring of ear veins under an inverted microscope. Animals received one of the following: an intraperitoneal, intravenous bolus, or intravenously infused dose of RB. An ear vein was illuminated by a green laser, and formation of a thrombus was captured with a digital camera. Only continuous intravenous infusion produced a steady-state RB plasma level and reproducible thrombus responses in different ear veins of the same rat. This system was then used to study the thrombogenic effects of iv-administered positively or negatively charged 60-nm ultrafine polystyrene particles (PSP). Significant dose-dependent enhancement of thrombus formation was found, as indicated by decreased laser illumination time to 33% of baseline values at 0.5 mg/kg. Negatively charged PSP of the same size failed to affect thrombus formation. We also studied the thrombogenic effect of PSP without the use of RB. The findings were the same as with RB, although the illumination time had to be increased. When 0.5 mg/kg was instilled intratracheally, the laser illumination time to form a thrombus was decreased to 42% of the baseline value, suggesting translocation of UFP into the bloodstream. These results are consistent with previous findings using the invasive model, and they validate the use of this non-invasive ear vein model to evaluate thrombogenic effects of UFP deposition in the respiratory tract.