CD63 tetraspanin slows down cell migration and translocates to the endosomal-lysosomal-MIICs route after extracellular stimuli in human immature dendritic cells

We analyzed herein whether members of the tetraspanin superfamily are involved in human immature dendritic cell (DC) functions such as foreign antigen internalization, phagocytosis, and cell migration. We show that CD63, CD9, CD81, CD82, and CD151 are present in immature DCs. Whereas CD9 and CD81 are mostly expressed at the cell surface, CD63 and CD82 are also located in intracellular organelles. Complexes of monoclonal antibody (Mab) FC-5.01-CD63 or Fab-5.01-CD63 were rapidly translocated "outside-in" and followed the endocytic pathway through early endosomes and lysosomes, reaching major histocompatibility complex (MHC) class II-enriched compartments (MIICs) in less than one hour. Internalization of CD63 was also observed during Saccharomyces cerevisiae phagocytosis. Moreover, an association of CD63 with the beta-glycan receptor dectin-1 was observed. Mabs against CD9, CD63, CD81, and CD82 enhanced by 50% the migration induced by the chemokines macrophage inflammatory protein-5 (MIP-5) and MIP-1alpha. Concomitantly, Mabs against CD63 and CD82 diminished the surface expression of CD29, CD11b, CD18, and alpha5 integrins. By immunoprecipitation experiments we found that CD63 associated with integrins CD11b and CD18. These results suggest that CD9, CD63, CD81, and CD82 could play a role in modulating the interactions between immature DCs and their environment, slowing their migratory ability. However, only CD63 would intervene in the internalization of complex antigens.     

Activities of several classes of acyclic nucleoside phosphonates against camelpox virus replication in different cell culture models

Camelpox virus (CMLV) is the closest known virus to variola virus. Here we report on the anti-CMLV activities of several acyclic nucleoside phosphonates (ANPs) related to cidofovir [(S)-1-(3-hydroxy-2-phosphonomethoxypropyl)cytosine (HPMPC; Vistide)] against two CMLV strains, CML1 and CML14. Cytopathic effect (CPE) reduction assays performed with human embryonic lung fibroblast monolayers revealed the selectivities of the first two classes of ANPs (cHPMPA, HPMPDAP, and HPMPO-DAPy) and of the hexadecyloxyethyl ester of 1-{[(5S)-2-hydroxy-2-oxido-1,4,2-dioxaphosphinan-5-yl]methyl}-5-azacytosine (HDE-cHPMP-5-azaC), belonging to the newly synthesized ANPs, which are HPMP derivatives containing a 5-azacytosine moiety. The inhibitory activities of ANPs against both strains were also confirmed with primary human keratinocyte (PHK) monolayers, despite the higher toxicity of those molecules on growing PHKs. Virus yield assays confirmed the anti-CML1 and anti-CML14 efficacies of the compounds selected for the highest potencies in CPE reduction experiments. Ex vivo studies were performed with a 3-dimensional model of human skin, i.e., organotypic epithelial raft cultures of PHKs. It was ascertained by histological evaluation, as well as by virus yield assays, that CMLV replicated in the human skin equivalent. HPMPC and the newly synthesized ANPs proved to be effective at protecting the epithelial cells against CMLV-induced CPE. Moreover, in contrast to the toxicity on PHK monolayers, signs of toxicity in the differentiated epithelium were seen only at high ANP concentrations. Our results demonstrate that compounds belonging to the newly synthesized ANPs, in addition to cidofovir, represent promising candidates for the treatment of poxvirus infections.     

Effects of moderate beer consumption on blood lipid profile in healthy Spanish adults

Background and aimsTo analyse the association of moderate beer consumption on the blood lipid profile in healthy Spanish adults.Methods and resultsThe study had an intervention longitudinal design in which each subject established their own control with a previous wash-out phase. After a 30-day alcohol abstinence period, 57 healthy volunteers were submitted to a daily moderate intake of beer for 30 days. Serum total cholesterol, HDL-cholesterol, triacylglycerols, GOT, GPT, GGT and glucose values, as well as blood erythrocytes, haemoglobin, haematocrit and MCV levels, together with anthropometric parameters were determined at the beginning of the study (baseline levels) (a), after 1 month of alcoholic abstinence (b) and after 1 month of moderate beer consumption (c). Dietary intake was assessed twice by a 7-day dietary record.HDL-cholesterol, erythrocytes, haematocrit and MCV levels increased significantly (p < 0.05) after moderate beer consumption in women. In men, a decrease in HDL-cholesterol levels was observed after alcohol abstention. Haematocrit and MCV counts also increased significantly (p < 0.05) in men after moderate beer consumption. There were no dietary changes during the study.ConclusionIn healthy Spanish adults, the effects of moderate beer consumption during 1 month were associated with favourable changes on the blood lipid profile.     

Absence of myocardial dysfunction during cold pressor stress echocardiography in patients with endothelial dysfunction

Background: Several methods are available for the assessment of coronary endothelial function, but there are no reports to date regarding the usefulness of cold pressor stress echocardiography (CPSE). Objective: To assess regional systolic and diastolic left ventricular function using CPSE in patients with endothelial dysfunction. Methods: We studied 24 patients, of whom 10 were men, aged 27 to 68 years, who had coronary risk factors and a normal exercise MP-SPECT test. They were compared with 10 normal subjects (6 men), aged 21 to 44 years. All patients underwent a CPSE. Results: The cold pressor-MP-SPECT revealed myocardial ischemia in 10 patients (Group I) and was normal in 14 patients (Group II). All normal subjects (Group III) had normal cold pressor-MP-SPECT. The cold pressor test caused a significant increase in systolic BP in the three groups (baseline 117 ± 17 mmHg vs. postcold test 137 ± 16 mmHg, P < 0.05), without changes in heart rate, PR interval, or the corrected QT interval. During the CPSE, no patient developed WMA in 2D echo or changes in regional systolic or diastolic LV function in the pulsed Doppler tissue imaging. Conclusions: In patients with endothelial dysfunction and no known coronary artery disease, the ischemic response to the cold pressor-MP-SPECT is not accompanied by WMA or changes in regional systolic or diastolic LV function during CPSE. Such negative findings indicate that the amount of ischemia that occurs secondarily to endothelial dysfunction does not involve sufficient myocardial mass to cause contractile dysfunction.     

Autoimmune disease in families with multiple sclerosis: a population-based study

BACKGROUND: Evidence of an association between multiple sclerosis (MS) and other autoimmune diseases would substantiate the hypothesis that MS is an autoimmune disease, and implicate a common mechanism. We aimed to investigate and compare the rate of autoimmune disease in MS patients, in their first-degree relatives, and in their unrelated spouses. METHODS: We used data from a national, multicentre, population-based sample to investigate the rate of autoimmune disease in 5031 MS patients, 30 259 of their first-degree relatives, and 2707 spousal controls. We asked patients and controls whether they had any of ten autoimmune diseases: Crohn's disease, ulcerative colitis, rheumatoid arthritis, type 1 diabetes, psoriasis, pernicious anaemia, systemic lupus erythematosus, autoimmune thyroid disease, vitiligo, and myasthenia gravis. MS probands were also asked whether their first-degree relatives had Crohn's disease, ulcerative colitis, rheumatoid arthritis, or type 1 diabetes. FINDINGS: After correction for age and sex, we did not identify any increased risk of autoimmune diseases in MS patients compared with their spousal controls (odds ratio [OR]=1.07, 95% CI 0.86-1.23, chi(2)=0.47, p=0.49), or in the first-degree relatives of MS probands compared with controls (OR=0.89, 0.63-1.17, chi(2)=1.11, p=0.29). However, the reported frequency of autoimmune diseases did differ according to the sex of the interviewee (female vs male patients chi(2)=92.2, p<0.0001; female vs male spousal controls chi(2)=87.1, p<0.0001). MS patients had slightly higher rates of thyroid disease and pernicious anaemia than did controls, which is consistent with MHC associations for these diseases, but this effect disappeared when results were adjusted for sex. For eight other diseases the rates were similar in MS patients and controls. Families with multiple cases of MS were no more likely to report autoimmune diseases than families with single MS cases. INTERPRETATION: When data were adjusted for sex, no excess of common autoimmune diseases could be identified in MS patients or their families, including multicase pedigrees. Our results suggest that women are more aware of family medical histories than men, which emphasises the potential for ascertainment bias in unstratified data for a sex-limited disease. Family histories should thus be taken from male patients in the presence of a spouse.     

Conceptualizing impulsivity and risk taking in bipolar disorder: importance of history of alcohol abuse

Background:  Elevated levels of impulsivity and increased risk taking are thought to be core features of both bipolar disorder (BD) and addictive disorders. Given the high rates of comorbid alcohol abuse in BD, alcohol addiction may exacerbate impulsive behavior and risk-taking propensity in BD. Here we examine multiple dimensions of impulsivity and risk taking, using cognitive tasks and self-report measures, in BD patients with and without a history of alcohol abuse.
Methods:  Thirty-one BD subjects with a prior history of alcohol abuse or dependence (BD-A), 24 BD subjects with no history of alcohol abuse/dependence (BD-N), and 25 healthy control subjects (HC) were assessed with the Barratt Impulsiveness Scale (BIS) and the computerized Balloon Analogue Risk Task (BART).
Results:  Both BD groups scored significantly higher than controls on the BIS. In contrast, only the BD-A group showed impaired performance on the BART. BD-A subjects popped significantly more balloons than the BD-N and HC groups. In addition, subjects in the BD-A group failed to adjust their performance after popping balloons. Severity of mood symptomatology was not associated with performance on either task.
Discussion:  The current study supports a primary role of prior alcohol abuse in risk-taking propensity among patients with bipolar disorder. In addition, findings suggest that impulsivity and risky behavior, as operationalized by self-report and experimental cognitive probes, respectively, are separable constructs that tap distinct aspects of the bipolar phenotype.     

Acupuncture stimulation at Sanyinjiao: effect on the sodium pertechnetate bioavailability in rats

In clinical acupuncture, when acupuncture points are stimulated, several types of reflex responses can be evoked. Consequently, different categories of physiological responses are induced, which include changes in the activities of internal organs and tissues. The acupuncture point Sanyinjiao (SP6) has been used successfully to treat different human gastrointestinal conditions. The aim of this work was to investigate the effects of end-organ response induced by acupuncture point SP6 on the bioavailability of the radiopharmaceutical sodium pertechnetate (Na99mTcO4) in Wistar rats. Healthy rats were allocated into 2 groups, control-CG and treated-TG. TG was bilaterally stimulated at acupuncture point SP6 with stainless steel needles. Ocular plexus administration of Na99mTcO4 (3.7MBq) was carried out 10 min after every needle insertion in all animals. Ten minutes later, the animals were killed, the organs were isolated, the radioactivity was determined in a well gamma counter, and the percentage of injected dose per gram of tissue (%ID/g) was determined for each organ. The %ID/g was significantly altered (p < 0.05) in the small intestine of TG (0.56 +/- 0.09) when compared to CG (0.82 +/- 0.18). These results may suggest that this stimulation might induce physiological responses capable of altering the bioavailability of the radiopharmaceutical sodium pertechnetate. These findings aid in providing a better understanding of acupuncture and its effects on various organs and tissues.     

Sertraline and Citalopram Actions on Gut Barrier Function

Introduction

Disruption of intestinal barrier is a key component to various diseases. Whether barrier dysfunction is the cause or effect in these situations is still unknown, although it is believed that translocation of luminal content may initiate gastrointestinal or systemic inflammatory disorders. Since trauma- or infection-driven epithelial permeability depends on Toll-like receptor (TLR) activity, inhibition of TLR signaling has been proposed as a strategy to protect intestinal barrier integrity after infection or other pathological conditions. Recently, selective serotonin recapture inhibitors including sertraline and citalopram were shown to inhibit TLR-3 activity, but the direct effects of these antidepressant drugs on the gut mucosa barrier remain largely unexplored.

Materials and methods

To investigate this, two approaches were used: first, ex vivo studies were performed to evaluate sertraline and citalopram-driven changes in permeability in isolated intestinal tissue. Second, both compounds were tested for their preventive effects in a rat model of disrupted gut barrier, induced by a low protein (LP) diet.

Results

Only sertraline was able to increase transepithelial electrical resistance in the rat colon both when used in an ex vivo (0.8 μg/mL, 180 min) or in vivo (30 mg/kg p.o., 20 days) fashion. However, citalopram (20 mg/kg p.o., 20 days), but not sertraline, prevented the increase in phospho–IRF3 protein, a marker of TLR-3 activation, in LP-rat ileum. Neither antidepressant affected locomotion, anxiety-like behaviours or stress-induced defecation.

Conclusion

Our data provides evidence to support the investigation of sertraline as therapeutic strategy to protect intestinal barrier function under life-threatening situations or chronic conditions associated with gut epithelial disruption.

     

Neonatal exposure to monosodium glutamate induces morphological alterations in suprachiasmatic nucleus of adult rat

Neonatal exposure to monosodium glutamate (MSG) induces circadian disorders in several physiological and behavioural processes regulated by the suprachiasmatic nucleus (SCN). The objective of this study was to evaluate the effects of neonatal exposure to MSG on locomotor activity, and on morphology, cellular density and expression of proteins, as evaluated by optical density (OD), of vasopressin (VP)‐, vasoactive intestinal polypeptide (VIP)‐ and glial fibrillary acidic protein (GFAP)‐immunoreactive cells in the SCN. Male Wistar rats were used: the MSG group was subcutaneously treated from 3 to 10 days of age with 3.5 mg/g/day. Locomotor activity was evaluated at 90 days of age using ‘open‐field’ test, and the brains were processed for immunohistochemical studies. MSG exposure induced a significant decrease in locomotor activity. VP‐ and VIP‐immunoreactive neuronal densities showed a significant decrease, while the somatic OD showed an increase. Major axes and somatic area were significantly increased in VIP neurons. The cellular and optical densities of GFAP‐immunoreactive sections of SCN were significantly increased. These results demonstrated that newborn exposure to MSG induced morphological alterations in SCN cells, an alteration that could be the basis for behavioural disorders observed in the animals.