Biased t cell receptor v gene usage in rheumatoid arthritis. oligoclonal expansion of t cells expressing vα2 genes in synovial fluid but not in peripheral blood

Objective. To analyze the T cell receptor (TCR) variable (V) region gene usage in the rheumatoid joint. Methods. Monoclonal antibodies (MAb) were used to determine the prevalence of selected V elements on T cells in synovial fluid (SF) from rheumatoid arthritis (RA) patients and in peripheral blood (PB) from RA patients and normal controls. V_α2-positive PB and SF T cells from 1 patient were cloned by immediated limiting-dilution and analyzed by restriction mapping. Results. In 9 of 14 RA patients, SF was enriched in at least 1 of the selected V elements, compared with PB. TCR genes of the V_α2 family were the most frequently overrepresented in the SF (4 patients). The expanded V_α2-positive cells were oligoclonal in SF but heterogeneic in PB. Conclusion. Our data showing biased and clonally restricted TCR elements in the rheumatoid joint indicate major histocompatibility complex–restricted antigen recognition, rather than a “superantigen,” in the pathogenesis of RA.     

Metabolism and Health Effects of Rare Sugars in a CACO-2/HepG2 Coculture Model

Non-alcoholic fatty liver disease (NAFLD) has become the most prevalent liver disease worldwide and is impacted by an unhealthy diet with excessive calories, although the role of sugars in NAFLD etiology remains largely unexplored. Rare sugars are natural sugars with alternative monomers and glycosidic bonds, which have attracted attention as sugar replacers due to developments in enzyme engineering and hence an increased availability. We studied the impact of (rare) sugars on energy production, liver cell physiology and gene expression in human intestinal colorectal adenocarcinoma (Caco-2) cells, hepatoma G2 (HepG2) liver cells and a coculture model with these cells. Fat accumulation was investigated in the presence of an oleic/palmitic acid mixture. Glucose, fructose and galactose, but not mannose, l-arabinose, xylose and ribose enhanced hepatic fat accumulation in a HepG2 monoculture. In the coculture model, there was a non-significant trend (p = 0.08) towards higher (20–55% increased) median fat accumulation with maltose, kojibiose and nigerose. In this coculture model, cellular energy production was increased by glucose, maltose, kojibiose and nigerose, but not by trehalose. Furthermore, glucose, fructose and l-arabinose affected gene expression in a sugar-specific way in coculture HepG2 cells. These findings indicate that sugars provide structure-specific effects on cellular energy production, hepatic fat accumulation and gene expression, suggesting a health potential for trehalose and l-arabinose, as well as a differential impact of sugars beyond the distinction of conventional and rare sugars.     

A systematic review of hearing and vestibular function in carriers of the Pro51Ser mutation in the COCH gene

European Archives of Oto-Rhino-Laryngology - The Pro51Ser (P51S) COCH mutation is characterized by a late-onset bilateral sensorineural hearing loss (SNHL) and progressive vestibular deterioration....     

Susceptibility to sensitization. I. Sex differences in the enduring effects of chronic D-amphetamine treatment on locomotion, stereotyped behavior and brain monoamines

There are sex differences in a number of behavior elicited by amphetamine (AMPH). The purpose of the present experiment was to determine if there are also sex differences in the sensitization of the locomotor activity and stereotypy produced by repeated intermittent AMPH treatment, and whether this is accompanied by sex differences in dopamine (DA) metabolism. It was found that female rats showed greater and more rapid sensitization of locomotor activity and stereotyped behavior than males. In addition, prior exposure to AMPH was associated with an elevation in resting striatal dihydroxyphenlacetic acid (DOPAC) to DA ratios in female, but not male rats, suggesting a sex difference in one neurochemical correlate of sensitization. As a group, males were more variable and heterogeneous in their response to repeated AMPH treatment, because they were divisible into two neurochemically distinct subgroups on the basis of their change in behavior and females were not. This heterogeneity may make it more difficult to identify neurochemical correlates of sensitization in males. It is suggested that there is a sex difference in the responsiveness of brain DA systems to repetitive activation, and this contributes to individual variation in the susceptibility to sensitization.     

18F-labeled FECNT: a selective radioligand for PET imaging of brain dopamine transporters

Fluorine-18 labeled 2beta-carbomethoxy-3beta-(4-chlorophenyl)-8-(2-fluoroethyl)nort ropane (FECNT) was synthesized in the development of a dopamine transporter (DAT) imaging ligand for positron emission tomography (PET). The methods of radiolabeling and ligand synthesis of FECNT, and the results of the in vitro characterization and in vivo tissue distribution in rats and in vivo PET imaging in rhesus monkeys of [18F]FECNT are described. Fluorine-18 was introduced into 2beta-carbomethoxy-3beta-(4-chlorophenyl)-8-(2-fluoroethyl)nort ropane (4) by preparation of 1-[18F]fluoro-2-tosyloxyethane (2) followed by alkylation of 2beta-carbomethoxy-3beta-(4-chlorophenyl)nortropane (3) in 21% radiochemical yield (decay corrected to end of bombardment [EOB]). Competition binding in cells stably expressing the transfected human DAT serotonin transporter (SERT) and norepinephrine transporter (NET) labeled by [3H]WIN 35428, [3H]citalopram, and [3H]nisoxetine, respectively, indicated the following order of DAT affinity: GBR 12909 > CIT > 2beta-carbomethoxy-3beta-(4-chlorophenyl)-8-(3-fluoropropyl) nortropane (FPCT) > FECNT. The affinity of FECNT for SERT and NET was 25- and 156-fold lower, respectively, than for DAT. Blocking studies were performed in rats with a series of transporter-specific agents and demonstrated that the brain uptake of [18F]FECNT was selective and specific for DAT-rich regions. PET brain imaging studies in monkeys demonstrated high [18F]FECNT uptake in the caudate and putamen that resulted in caudate-to-cerebellum and putamen-to-cerebellum ratios of 10.5 at 60 min. [18F]FECNT uptake in the caudate/putamen peaked in less than 75 min and exhibited higher caudate- and putamen-to-cerebellum ratios at transient equilibrium than reported for 11C-WIN 35,428, [11C]CIT/RTI-55, or [18F]beta-CIT-FP. Analysis of monkey arterial plasma samples using high performance liquid chromatography determined that there was no detectable formation of lipophilic radiolabeled metabolites capable of entering the brain. In equilibrium displacement experiments with CIT in rhesus monkeys, radioactivity in the putamen was displaced with an average half-time of 10.2 min. These results indicate that [18F]FECNT is a radioligand that is superior to 11C-WIN 35,428, [11C]CIT/RTI-55, [18F]beta-CIT-FP, and [18F]FPCT for mapping brain DAT in humans using PET.     

Patient-related risk factors for requiring surgical intervention following a failed injection for the treatment of medial and lateral epicondylitis

Objectives: To identify risk factors for failure of a therapeutic injection leading to operative management of both medial and lateral epicondylitis.

Methods: A national database was used to query Medicare Standard Analytic Files from 2005–2012 for patients treated with therapeutic injections for medial or lateral epicondylitis using CPT codes for injections associated with corresponding ICD-9 diagnostic codes (726.31 and 726.32, respectively). Those who subsequently underwent surgical treatment following injection were identified. A multivariate binomial logistic regression analysis was utilized to evaluate patient-related risk factors for requiring surgery within 2 years after therapeutic injection.

Results: 1,837 patients received therapeutic injections for medial epicondylitis. 52 (2.8%) required ipsilateral surgery at a mean of 429 ± 28 days post-injection. Risk factors for requiring surgical intervention included age <65, obesity, and morbid obesity. 6,561 patients received therapeutic injections for lateral epicondylitis. 201 (3.1%) required subsequent surgery at a mean of 383 ± 128 days’ post-injection. Risk factors included age <65, tobacco use, diabetes mellitus and peripheral vascular disease.

Conclusion: The incidence of surgical intervention following a failed therapeutic injection for medial or lateral epicondylitis is low (~3%). Risk factors for failing a therapeutic injection include age <65 years and obesity (BMI > 30) for medial epicondylitis and age <65 years, smoking, diabetes mellitus and peripheral vascular disease for lateral epicondylitis. Patients with these identified risk factors presenting with medial or lateral epicondylitis should be cautioned that they carry a higher risk of subsequent surgical treatment. Level of Evidence: Therapeutic, III.     

Blood donation leads to a decrease in natural killer cell activity: a study in normal blood donors and cancer patients

Transfusion-induced immunosuppression has long been known to be beneficial for organ transplantation patients, but recent retrospective studies suggest that blood transfusions may be detrimental for patients with cancer. If autologous blood is used to avoid immunosuppression, the assumption is that the procedure, involving blood donation, is immunologically neutral. In the present study, this assumption was evaluated by monitoring 33 normal blood donors and 16 colorectal cancer patients before and after donation of 1 (500 mL) and 2 units of blood, respectively. The cancer patients belonged to the autologous arm of a randomized trial in which the effects of allogeneic versus autologous blood on cancer prognosis were studied. The patients donated 2 units of blood with an interval of 3 to 4 days between donations. Flow cytometric analysis revealed that blood donation by normal donors and cancer patients had no effect on the proportion of B, T, and natural killer (NK) cells. Only the total number of lymphocytes was significantly decreased in the normal donors on Day 12 after donation. Blood donation had no significant effect on T-cell function assessed by phytohemagglutinin stimulation in normal donors or in cancer patients donating 2 units of blood. A significant depression of NK cell function (88% and 74% of predonation levels) was observed in normal donors on Days 2 and 5 after donation; on Day 12, the activity was again normal. Colorectal cancer patients had a significantly depressed NK cell activity (54% of predonation activity) on Day 12 after the first donation.(ABSTRACT TRUNCATED AT 250 WORDS)     

In vitro leishmanicidal activity of Tityus discrepans scorpion venom

Leishmania parasites are sensitive to peptides with antimicrobial and ion-channel inhibitory activity. Because scorpion venoms are rich sources of such peptides, the leishmanicidal effect of Tityus discrepans venom was investigated. A negative correlation between cell growth and venom concentration was observed for venom-treated cultures of Leishmania (L.) mexicana mexicana promastigotes; 50% growth inhibition was obtained at 0.4 μg/ml. Light microscopy showed rounded, highly vacuolated L. (L.) m. mexicana cells with impaired flagellar motion after 15 min of incubation at 35 μg/ml. Ultrastructural studies confirmed an intense cytoplasm vacuolation and also enlargement of the flagellar pocket. Survival rates for New World Leishmania promastigotes (75% venom effective concentration, μg/ml) obtained after acute (1 h) venom toxicity tests were: L. (L.) m. mexicana (2.3), Leishmania (V.) braziliensis (11.3), and Leishmania (L.) chagasi (56.2). Heat (90°C) treatment of venom and fraction TdII abolished most of their leishmanicidal effect. Acute toxicity assays performed with Sephadex G-50 fractions indicated that leishmanicidal activity is associated with the venom lowest molecular mass components (2.8–7.4 kDa), as determined by MALDI-TOF mass spectrometry.