Multispecies probiotic protects gut barrier function in experimental models

AIM:To investigate the effect of the probiotic combination Lactibiane Tolerance?(LT)on epithelial barrier function in vitro and in vivo.METHODS:The effect of the multispecies probiotic LT was assessed on several models of epithelial barrier function both in vitro(in basal and inflammatory conditions)and in vivo[visceral hypersensitivity induced by chronic stress or by colonic perfusion of a fecal supernatant(FSN)from patients with irritable bowel syndrome(IBS)].In vitro,we measured the permeability of confluent T84 cell monolayers incubated with or without LT by evaluating the paracellular flux of macromolecules,in basal conditions and after stimulation with lipopolysaccharide(LPS)or with conditioned medium of colonic biopsies from IBS patients(IBS-CM).In vivo,male C57/Bl6 mice received orally NaCl or LT for 15 d and were submitted to water avoidance stress(WAS)before evaluating visceral sensitivity by measuring the myoelectrical activity of the abdominal muscle and the paracellular permeability with 51Cr-EDTA.Permeability and sensitivity were also measured after colonic instillation of FSN.Tight-junctions were assessed by immunoblotting and TLR-4 expression was evaluated by immunohistochemistry RESULTS:Incubation of T84 cell monolayers with LT in basal conditions had no significant effect on permeability(P>0.05 vs culture medium).By contrast,addition of LT bacterial bodies(LT)completely prevented the LPS-induced increase in paracellular permeability(P<0.01 vs LPS 10 ng/mL(LPS 10);P<0.01 vs LPS 100ng/mL(LPS 100),P>0.05 vs culture medium).The effect was dose dependent as addition of 109 LT bacterial bodies induced a stronger decrease in absorbance than 106 LT(109 LT+LPS 10:-20.1%±13.4,P<0.01vs LPS 10;106 LT+LPS 10:-11.6%±6.2,P<0.01 vs LPS 10;109 LT+LPS 100:-14.4%±5.5,P<0.01 vs LPS 100;106 LT+LPS 100:-11.6%±7.3,P<0.05 vs LPS 100).Moreover,the increase in paracellular permeability induced by culturing T84 cells with conditioned medium of colonic biopsies from IBS patients(IBS-CM)was completely inhibited in the presence of 109 LT(P<0.01 vs IBS-CM).LT also significantly prevented the epithelial disruption induced by intracolonic infusion of fecal supernatant from IBS patients(P<0.01 vs IBS FSN)or water avoidance stress P<0.01 vs WAS)in C57/Bl6 mice and increased the expression of occludin in vitro and in vivo,as assessed by immnunoblotting.The WAS-induced effect on visceral sensitivity was prevented by LT treatment since values obtained for all steps of colorectal distension were significantly(P<0.01)different from the WAS group.Finally,LT downregulated the response mediated through TLR-4 in vitro(decrease in tumor necrosis factorαsecretion in response to LPS:-65.8%for 109 LT and-52.5%for 106LT,P<0.01 vs LPS)and in vivo(inhibition of WAS induced an increase in TLR-4 expression in the LT treated mice colon,P<0.01 vs WAS).CONCLUSION:The probiotic LT mix prevented the disruption to the epithelial barrier induced by LPS,stress or colonic soluble factors from IBS patients and prevented visceral hypersensitivity.     

Abnormal inhibin A and inhibin B secretion in obese women with and without insulin resistance.

AIM: The present study aimed to establish inhibin A and B serum levels during the menstrual cycle of obese women, and its usefulness as an index of luteal-phase follicular development. MATERIALS AND METHODS: Twenty-one obese patients (mean body mass index: 34.9+/-3.7 kg/m2; range: 30.0-45.0 kg/m2) were submitted to basal hormonal measurements and an oral glucose tolerance test after challenge with 75 g glucose. Progesterone and inhibin A and B levels were determined 5-7 days after the menstrual cycle and 7 days prior to expected menses. RESULTS: As expected, an increase in inhibin A and a decrease in inhibin B were observed when first-phase samples were compared with samples obtained after 15-20 days. On the other hand, the percentage variation of both inhibin A and B was at least four times smaller than the values for normal women described previously by other authors employing the same enzyme-linked immunosorbent assays. A small number of obese women presented ovulatory cycles characterized by progesterone concentration higher than 5.8 ng/ml. The percentage elevation (>190%) of inhibin A in the second samples was in agreement with the progesterone levels, but it seemed to be more sensitive for the detection of follicle luteinization. CONCLUSION: We conclude that obese women present less percentage variation of both inhibin A and B during the menstrual cycle, associated with a low frequency of ovulatory cycles. In obese women, the percentage increase of inhibin A can represent an additional marker to recognize follicle luteinization.     

Purification, physicochemical properties, thermal analysis and antinociceptive effect of atranorin extracted from Cladina kalbii

Lichens and their secondary metabolites have attracted the interest of many researchers. Some species have been shown to contain substances with remarkable biologic activity, as antimicrobial, mainly against Gram positive bacteria; antineoplasic acting on solid and ascetic tumors, or in culture cells; antiviral; hypotensive; and spasmolytic effects. The aim of this study was to isolate and characterize atranorin, one of the major constituents which presents in Cladina kalbii (DES ABB.) AHTI., and analyze its antinociceptive effect. The antinociceptive activity was verified in acetic acid-induced writhing test and formalin test with mice. In this work it was observed that atranorin was effective in significant reducing (p<0.05) abdominal writhing at doses of 200 and 400 mg/kg (p.o.) by 52.6 and 61.3%, respectively, when compared to control group (vehicle). The formalin test showed in 200 and 400 mg/kg (p.o.) that atranorin injection was able to inhibit the inflammatory processes (second phase) dose dependently.     

Prospects for Personalised Treatment of Patients with Radioiodine-avid Locally Recurrent or Metastatic Thyroid Cancer

Although most patients with metastatic or inoperable locally recurrent differentiated thyroid cancer have radioiodine-avid disease, the outcome for patients who do not achieve remission with radioiodine therapy is poor. Most centres use fixed empirical activities of radioiodine to treat these patients, which is in contrast to other areas of oncology, where there is a shift to more individualised treatment. The use of dosimetry techniques to calculate a more appropriate activity of radioiodine for each patient may increase the effectiveness of radioiodine therapy but is more complex, time-consuming and of unproven benefit. This review addresses some of the limitations of empirical radioiodine therapy, discusses existing dosimetry-based approaches to individualising therapy and proposes further work in this area. A prospective randomised controlled trial comparing empirical activities of radioiodine with activities guided by a combination of lesional dosimetry and maximum safe dose has not been carried out previously. Although considerable challenges in the design of such a study remain, a network of centres in the UK now has the potential to take this forward.     

Prestorage leukoreduction and low-temperature filtration reduce hemolysis of stored red cell concentrates

BACKGROUND: Universal prestorage leukoreduction in Canada created the perception that stored red cells (RBCs) are more hemolyzed than their unfiltered predecessors. A pool-split design tested the effects of leukoreduction on hemolysis of stored RBCs. STUDY DESIGN AND METHODS: Two ABO-matched units were pooled, divided, and then processed into leukoreduced (LR) and nonleukoreduced (NLR) units with the Pall LT-WB or RC-PL systems and sampled during standard processing and storage for testing of sterility, counts, hemolysis, and osmotic fragility. RESULTS: Room temperature (RT) filtration of 10 pairs of LT-WB-LR and -NLR units showed significantly different percentage of hemolysis (0.39%) and osmotic fragility (0.643%) at 42 days. Cold-stored and -filtered units (2 days at 4 degrees C before processing) were less hemolyzed, but showed a similar proportional decrease of hemolysis in LR units (0.13% vs. 0.25% at 42 days). RBCs from RC-PL systems showed the lowest hemolysis although there was a filtration effect (0.05% vs. 0.12%, 42 days). Osmotic fragility paralleled hemolysis. Segment samples gave inaccurate results. Two-day prefiltration cold storage reduced hemolysis from 0.36 to 0.07 percent (42 days, p < 0.001). RT-LR hemolysis became significantly higher by Day 10 and 4 degrees C LR by Day 12. NLR units showed hemolysis by Day 7. LR units filtered cold were less hemolyzed (p < 0.05) than RT-LR but osmotic fragility was unchanged. CONCLUSIONS: LR-RBCs prepared by any of three methods (LT-WB, RT or cold; RC-PL), filtered at 4 degrees C, were less hemolyzed during storage than nonfiltered concentrates: 4 degrees C leukoreduction is beneficial for RBCs and does not cause hemolysis or enhance fragility.