Evaluation of the sensitivity of Moroccans to shrimp tropomyosin and effect of heating and enzymatic treatments

The aim of this work was to evaluate the IgE-sensitivity to shrimp tropomyosin (ST) in a Moroccan population from Fez region, and then to study the effect of temperature and enzymatic digestion on the allergenicity of ST. This work was conducted with a questionnaire completed by a sera-bank, obtained from 500 patients recruited from Fez Hospitals. Their sera were analyzed for specific IgE-sensitivity to ST. From questionnaire, 9.8% reported allergy to fish and shellfish where shrimp was one of the most common species causing allergy in patients. Evaluation of specific IgE showed that 10.2% of patients present higher values. Further indirect ELISA and Dot-blot results indicated that ST showed a decrease in the human IgE binding under heating or pepsin hydrolysis. These results demonstrate that this population was sensitive to ST and the sensitivity could be reduced by heating and more where it was digested by pepsin.     

Gastric and masticatory performances in full‐arch immediate loading rehabilitated patients

Full‐arch immediate loading implant rehabilitations provide patients with compromised dentition an effective treatment to improve their aesthetic and function. Aim of this prospective cohort study was to investigate the correlation between masticatory ability and gastric emptying rates among these patients. Ten subjects (five men and five women) with compromised dentition were tested in two occasions: before treatment and 30 days after the immediate loading rehabilitation. Masticatory ability was evaluated using the sieves test, and the gastric half emptying time (T_1/2) was assessed by means of the ¹³C‐octanoic acid breath test. A statistically significant increment (P < 0·005) in masticatory ability was found only in reference to the particles smaller than or equal to 4·75 mm, whereas the gastric emptying rate showed a statistically significant reduction between pre‐ and post‐treatment (P = 0·003). A moderate negative correlation (rho = 0·64, P = 0·048) between the percentage change in masticatory ability and the percentage change in gastric emptying rate was evidenced. Patients with compromised dentition rehabilitated with full‐arch immediate implant prostheses present a significant improvement of the gastric process.     

Streptokinase may play role in expanding non-operative management of infected walled off pancreatic necrosis: Preliminary results

Goals and backgroundWe evaluated ex and in vivo effect of streptokinase on pancreatic necrosum to improve the success rate of pigtail catheter drainage and irrigation in infected walled off pancreatic necrosis using step up approach and also looked at potential risk of bleeding.Experiment and clinical cases1000 IU/ml of streptokinase was added to 10 g. of intra-operatively obtained fresh tissue of peripancreatic necrosis and results compared to treatment with saline. Mixture was incubated for 12 h in thermostat at 37.5 °C and subjected to histopathology. Subsequently streptokinase (50,000 units thrice a day for 5 days through PCD) was used in two patients with walled off pancreatic necrosis (WOPN) not responding to step up approach and who were being considered for surgery.Grossly there was fragmentation of necrosum in streptokinase treated tissue. Microscopically complete loss of supportive collagenous framework was noted in streptokinase treated necrosum with clumping of necrotic tissue into structure-less mass. No such changes were discernible in saline treated tissue. In two patients with WOPN there was clearance of debris after streptokinase instillation. None of the patients was on thromboprophylaxis and bleeding was not noticed in any of the patients.ConclusionBased on ex vivo effect of streptokinase in dissolution of necrosum at periphery, we believe that in patients with walled off pancreatic necrosis (WOPN) not responding to pigtail catheter drainage and saline irrigation; streptokinase may prove to be useful adjunct.     

Sweet potato leaf extract inhibits the simulated in vitro gastrointestinal digestion of native starch

Several studies have reported the therapeutic use of caffeoylquinic acid (CQA) derivatives in the management of hyperglycemia. This study used a simulated in vitro gastrointestinal digestion model to assess the inhibitory effects of CQA derivatives-rich sweet potato leaf extract (SPLE) and a commercially produced green coffee bean extract (GCBE), each with total polyphenols contents of 452 mg g⁻¹ and 278 mg g⁻¹, respectively, against starch digestion. The changes in the amounts of total polyphenols and total CQA derivatives during in vitro gastrointestinal digestion were also examined. The results indicated that both extracts contained substantial levels of CQA derivatives (136 mg g⁻¹ and 83.5 mg g⁻¹ of extract for SPLE and GCBE, respectively). The amounts of total polyphenols and total CQA derivatives in 20 mg of SPLE and GCBE samples decreased from 9.04 mg to 0.58 mg and from 5.56 mg to 0.58 mg, and from 2.72 mg to 0.16 mg and from 1.67 mg to 0.10 mg, respectively, following in vitro gastrointestinal digestion and subsequent dialysis. When SPLE and GCBE were accompanied with starch for in vitro digestion test, they both exhibited inhibitory effect against starch digestion during simulated intestinal digestion, with estimated half maximal inhibitory concentration (IC₅₀) values of 4.91 mg and 6.06 mg polyphenols, respectively. The amount of glucose permeated through dialysis membrane also decreased significantly in comparison with the extract-negative control. Thus, both SPLE and GCBE were capable of modulating the release of glucose from starch digestion in simulated intestinal tract. The observed inhibitory effects against glucose release were presumably due in part to the presence of CQA derivatives in the tested extracts. The SPLE had higher inhibitory effect against in vitro starch digestion than the commercially prepared reference GCBE. Therefore, the SPLE might be used to manage hyperglycemia over the long term.     

Biological effect of food additive titanium dioxide nanoparticles on intestine: an in vitro study

Titanium dioxide nanoparticles (TiO₂ NPs) are widely found in food‐related consumer products. Understanding the effect of TiO₂ NPs on the intestinal barrier and absorption is essential and vital for the safety assessment of orally administrated TiO₂ NPs. In this study, the cytotoxicity and translocation of two native TiO₂ NPs, and these two TiO₂ NPs pretreated with the digestion simulation fluid or bovine serum albumin were investigated in undifferentiated Caco‐2 cells, differentiated Caco‐2 cells and Caco‐2 monolayer. TiO₂ NPs with a concentration less than 200 µg ml^–1 did not induce any toxicity in differentiated cells and Caco‐2 monolayer after 24 h exposure. However, TiO₂ NPs pretreated with digestion simulation fluids at 200 µg ml^–1 inhibited the growth of undifferentiated Caco‐2 cells. Undifferentiated Caco‐2 cells swallowed native TiO₂ NPs easily, but not pretreated NPs, implying the protein coating on NPs impeded the cellular uptake. Compared with undifferentiated cells, differentiated ones possessed much lower uptake ability of these TiO₂ NPs. Similarly, the traverse of TiO₂ NPs through the Caco‐2 monolayer was also negligible. Therefore, we infer the possibility of TiO₂ NPs traversing through the intestine of animal or human after oral intake is quite low. This study provides valuable information for the risk assessment of TiO₂ NPs in food. Copyright © 2015 John Wiley & Sons, Ltd.     

Gastrointestinal melatonin: localization,function, and clinical relevance

The gastrointestinal tract of vertebrate species is a rich source of extrapineal melatonin. The concentration of melatonin in the gastrointestinal tissues surpasses blood levels by 10–100 times and there is at least 400× more melatonin in the gastrointestinal tract than in the pineal gland. The gastrointestinal tract contributes significantly to circulating concentrations of melatonin, especially during the daytime and melatonin may serve as an endocrine, paracrine, or autocrine hormone influencing the regeneration and function of epithelium, enhancing the immune system of the gut, and reducing the tone of gastrointestinal muscles. As binding sites for melatonin exhibit circadian variation in various species, it has been hypothesized that some melatonin found in the gastrointestinal tract might be of pineal origin. Unlike the photoperiodically regulated production of melatonin in the pineal, the release of gastrointestinal melatonin seems to be related to the periodicity of food intake. Phylogenetically, melatonin and its binding sites were detected in the gastrointestinal tract of lower vertebrates, birds, and mammals. Melatonin was found also in large quantities in the embryonic tissue of the mammalian and avian gastrointestinal tract. Food intake and, paradoxically, also long-term food deprivation resulted in an increase of tissue and plasma concentrations of melatonin. Melatonin release may have a direct effect on many gastrointestinal tissues but may also well influence the digestive tract indirectly, via the central nervous system and the sympathetic and parasympathetic nerves. Melatonin prevents ulcerations of gastrointestinal mucosa by an antioxidant action, reduction of secretion of hydrochloric acid, stimulation of the immune system, fostering epithelial regeneration, and increasing microcirculation. Because of its unique properties, melatonin could be considered for prevention or treatment of colorectal cancer, ulcerative colitis, gastric ulcers, irritable bowel syndrome, and childhood colic.     

Circadian regulation of epithelial functions in the intestine

Many physiological functions exhibit a diurnal rhythmicity that is influenced by biological clocks and feeding rhythms. In this review, we discuss the growing evidence showing the important role of circadian rhythms in regulating intestinal mucosa. First, we introduce the molecular timing system and the interrelationship between the master biological clock in the suprachiasmatic nuclei of the brain and the peripheral intestinal clock and provide evidence that the intestinal clock is entrained with the external environment. Second, we review the circadian rhythmicity of enterocyte proliferation and the largely unknown regulatory mechanisms behind these rhythms. Finally, we focus on the circadian clock control of food processing that functions by regulating the expression of digestive enzymes and intestinal nutrient and salt transporters. The concepts to be discussed highlight the ability of the intestinal epithelium to utilize self‐sustained clock signals together with signals associated with changes in the cellular environment and to use endogenous temporal control of the gastrointestinal functions to meet varying physiological and pathophysiological demands. The fact that internal de‐synchronizations within the body, such as those that occur in shift workers or with changes in food intake behaviour, are often associated with malfunctions of the gastrointestinal tract indicates that more information about the connections between the circadian clock and intestinal mucosa/transporting enterocytes could provide clues for future therapies.     

Forensic analysis of diatom in the spleen and heart in Sprague-Dawley rats

Post-mortem diagnosis of drowning is a real challenge, especially in a decomposed body. The diatom test is considered a reliable scientific assessment in several countries including China. The lungs, liver, and kidneys are usually used in the diatom test, whereas the spleen and heart are rarely reported. In our study, we set up five water samples with different concentrations of diatoms. One-hundred and twenty adult SD rats were employed, assigned to the Drowning Group (DG), Post-mortem Submersion Group (PSG), Drowning Control Group (DG-Control), and Post-mortem Submersion Control Group (PSG-Control). Rats in DG and PSG were submerged in water samples, respectively. After the microwave digestion and vacuum filtration method for diatom enrichment, the diatom numbers were counted. The results showed no diatom was found in either the DG-Control or PSG-Control group. Although significant differences in the diatoms number were found between the DG and PSG groups, the heart and spleen revealed no significant differences. These two organs equally had high specificity and low sensitivity. Based on our data, we concluded that the heart and spleen can be used for the diagnosis of drowning, and detection of diatoms in systemic circulation organs may be more suitable for the diatom test, especially when exogenous contamination is avoided.     

改良分次酶消化法分离培养兔肌腱干细胞及诱导分化鉴定

目的  采用改良分次酶消化法体外分离培养家兔肌腱干细胞,观察其生物学特性,并进行诱导分化及鉴定。方法  无菌条件下取出家兔髌腱组织,分别运用改良的分次酶消化法和酶消化后低密度稀释接种法进行分离、培养、传代,用倒置相差显微镜观察细胞形态特征并绘制两种方法的生长曲线,通过流式细胞鉴定仪检测肌腱干细胞表面抗原标志物的表达,取P3-P4代肌腱干细胞向成骨细胞、成软骨细胞诱导分化并鉴定。结果  改良的分次酶消化法较酶消化后低密度稀释接种法细胞增殖速度加快,形态均一,杂质细胞少。分离出的肌腱干细胞表面抗原标志CD90、CD44呈阳性,而CD34、CD14呈阴性,证实之前分离的细胞为肌腱干细胞。鉴定出肌腱干细胞具有向成骨细胞和成软骨细胞分化能力。结论  采用改良的分次酶消化法分离培养兔肌腱干细胞简单易行,肌腱干细胞生长及传代速度可观,活性良好,纯度较高。另外,肌腱干细胞的成功分离培养,也为肌腱相关疾病的研究开辟了一条新途径。

     

Reference antigen-free and antibody-free LTD-IDMS assay for influenza H7N9 vaccine in vitro potency determination

Influenza vaccines are the most effective intervention to prevent the substantial public health burden of seasonal and pandemic influenza. Hemagglutinin (HA), as the main antigen in inactivated influenza vaccines (IIVs), elicits functional neutralizing antibodies and largely determines IIV effectiveness. HA potency has been evaluated by single-radial immunodiffusion (SRID), the standard in vitro potency assay for IIVs, to predict vaccine immunogenicity with a correlation to protective efficacy. We previously reported that limited trypsin digestion (LTD) selectively degraded stressed HA, so that an otherwise conformationally insensitive biophysical quantification technique could specifically quantify trypsin-resistant, immunologically active HA. Here, we demonstrate that isotope dilution mass spectrometry (IDMS), a method capable of quantifying the absolute HA concentration without reference antigen use, can be further expanded by adding LTD followed with precipitation to selectively quantify the active HA. We test the LTD-IDMS assay on H7N9 vaccines stressed by low pH, raised temperature, or freeze/thaw cycles. This method, unlike SRID, has no requirement for strain-specific reference antigens or antibodies and can generate potency values that correlate with SRID. Thus, LTD-IDMS is a promising alternative in vitro potency assay for influenza vaccines to complement and potentially replace SRID in a pandemic when strain specific reagents may not be readily available.