Effect of Bacillus subtilis,Enterococcus faecium,and Enterococcus faecalis supernatants on serotonin transporter expression in cells and tissues

BACKGROUND Bacillus subtilis(B.subtilis),Enterococcus faecium(E.faecium),and Enterococcus faecalis(E.faecalis)are probiotics that are widely used in the clinical treatment of irritable bowel syndrome(IBS).Whether the supernatants of these three probiotics can improve gastrointestinal sensation and movement by regulating the serotonin transporter(SERT)expression needs to be clarified.AIM To investigate whether B.subtilis,E.faecium,and E.faecalis supernatants can upregulate SERT expression in vitro and in vivo.METHODS Caco-2 and HT-29 cells were stimulated with probiotic culture supernatants for 12 and 24 h,respectively.A male Sprague-Dawley rat model of post-infectious irritable bowel syndrome(PI-IBS)was established and the rats were treated with phosphate-buffered saline(group A)and three probiotics culture supernatants(groups B,C,and D)for 4 wk.The levels of SERT were detected by quantitative PCR and western blotting.RESULTS The levels of SERT at post-treatment 12 and 24 h were significantly elevated in Caco-2 cells treated with B.subtilis supernatant compared with those in the control group(aP<0.05).Those levels were markedly upregulated in Caco-2 cells stimulated with E.faecium and E.faecalis supernatants at 24 h(aP<0.05).In addition,SERT expression in groups B,C,and D was significantly higher than that in group A in the 2nd wk(aP<0.05).Increased SERT expression was only found in group D in the 3rd wk(aP<0.05).However,there was no significant difference in SERT expression between the groups in the last week(P>0.05).CONCLUSION The supernatants of B.subtilis,E.faecium,and E.faecalis can upregulate SERT expression in intestinal epithelial cells and the intestinal tissues in the rat model of PI-IBS.     

Investigation of Self-Healing Mortars with and without Bagasse Ash at Pre- and Post-Crack Times

Cracks in typical mortar constructions enhance water permeability and degrade ions into the structure, resulting in decreased mortar durability and strength. In this study, mortar samples are created that self-healed their cracks by precipitating calcium carbonate into them. Bacillus subtilus bacterium (10⁻⁷, 10⁻⁹ cells/mL), calcium lactate, fine aggregate, OPC-cement, water, and bagasse ash were used to make self-healing mortar samples. Calcium lactates were prepared from discarded eggshells and lactic acid to reduce the cost of self-healing mortars, and 5% control burnt bagasse ash was also employed as an OPC-cement alternative. In the presence of moisture, the bacterial spores in mortars become active and begin to feed the nutrient (calcium lactate). The calcium carbonate precipitates and plugs the fracture. Our experimental results demonstrated that cracks in self-healing mortars containing bagasse ash were largely healed after 3 days of curing, but this did not occur in conventional mortar samples. Cracks up to 0.6 mm in self-healing mortars were filled with calcite using 10⁻⁷ and 10⁻⁹ cell/mL bacteria concentrations. Images from an optical microscope, X-ray Diffraction (XRD), and a scanning electron microscope (SEM) were used to confirm the production of calcite in fractures. Furthermore, throughout the pre- and post-crack-development stages, self-healing mortars have higher compressive strength than conventional mortars. The precipitated calcium carbonates were primed to compact the samples by filling the void spaces in hardened mortar samples. When fissures developed in hardened mortars, bacteria became active in the presence of moisture, causing calcite to precipitate and fill the cracks. The compressive strength and flexural strength of self-healing mortar samples are higher than conventional mortars before cracks develop in the samples. After the healing process of the broken mortar parts (due to cracking), self-healing mortars containing 5% bagasse ash withstand a certain load and have greater flexural strength (100 kPa) than conventional mortars (zero kPa) at 28 days of cure. Self-healing mortars absorb less water than typical mortar samples. Mortar samples containing 10⁻⁷ bacteria cells/mL exhibit greater compressive strength, flexural strength, and self-healing ability. XRD and SEM were used to analyze mortar samples with healed fractures. XRD, FTIR, and SEM images were also used to validate the produced calcium lactate. Furthermore, the durability of mortars was evaluated using DTA-TGA analysis and water absorption tests.     

Current intravesical therapy for non-muscle invasive bladder cancer

Introduction: Transurethral resection of the bladder tumour (TURBT) is still the standard initial treatment for non-muscle invasive bladder cancer (NMIBC). However, even after a radical resection, recurrence (30 – 80%) and progression (1 – 45%) are commonly seen. Intravesical therapy provides direct contact of the agent with the bladder mucosa and clearly has improved the outcome, especially in high-risk disease.

Areas covered: The role of a good initial TURBT is emphasized. Risk assessment tools are discussed. Different intravesical therapies are enumerated according to the latest literature, with the emphasis on Bacillus Calmette–Guérin (BCG), including the discussion on the optimal dose and schedule. New developments are mentioned.

Expert opinion: A radical TURBT is essential for good prognosis. For optimal visualisation of tumours, fluorescence techniques should be used with low threshold, especially in case of suspicion of carcinoma in situ (CIS). Increased completeness of the resection will lead to less persisting disease and less need for adjuvant treatment. A re-TURBT should be done when in doubt of radical resection (judged by the pathologist or the surgeon). Risk assessment is essential, but the available tools are outdated. A single post-operative instillation (SPI) with chemotherapy is only indicated in low-risk disease. BCG is the treatment of choice for high-grade disease. BCG should be given as maintenance. Awareness of deterioration of the prognosis after progression is of great importance. In BCG failures, cystectomy should be strongly advised. Chemotherapy in combination with hyperthermia seems to be a new promising treatment.     

Attendance and vaccination at immunization clinics in rural Gambia before and during the COVID-19 pandemic

IntroductionThe COVID-19 pandemic has affected the delivery of essential health services, such as routine immunization. We assessed the impact of the pandemic on the uptake of routine immunization in rural Gambia.MethodsWe collected real-time vaccine administration data in the Basse and Fuladu West Health & Demographic Surveillance Systems from September 01, 2019, to December 31, 2020. We assessed the monthly number of Expanded Program on Immunization (EPI) clinic attendances and vaccines administered, comparing data during the baseline period (September 01, 2019–March 31, 2020), COVID-19 interruption period (April 01–June 30, 2020), initial recovery period (Jul 01–September 30, 2020) and the late recovery period (October 01–December 31, 2020).ResultsCompared to the baseline period, there was an overall average monthly decline of 13.4% in EPI attendance and 38.3% reduction in average monthly immunizations during the interruption period. This decrease was particularly noticeable for Bacille Calmette-Guérin (BCG) (47.2%), birth dose hepatitis B (Hep B) (46.9%), 1st dose pentavalent (Penta1) (43.1%), 1st dose pneumococcal conjugate vaccine (PCV1) (42.4%), and measles vaccines (15.5%). Comparing the late recovery to baseline period, average monthly EPI attendance was 5.3% higher, with 1.9% increase in average monthly immunizations. Monthly immunizations for BCG were 3.0% greater, 2.5% greater for Hep B, 22.7% greater for oral polio vaccine (OPV1), 2.0% less for Penta1, 19.2% less for Penta2, and 2.6% less for PCV1.ConclusionThe reduced EPI attendance during the pandemic interruption period lasted only 3 months. Significant recovery in EPI attendance occurred during the late recovery period, while rates of monthly immunization returned to pre-pandemic levels for most antigens. EPI programmes should implement strategies to deliver missed antigens when infants do present to EPI clinics, aware that missed doses may be age dependent.     

A 90-day safety study of genetically modified rice expressing Cry1Ab protein (Bacillus thuringiensis toxin) in Wistar rats.

An animal model for safety assessment of genetically modified foods was tested as part of the SAFOTEST project. In a 90-day feeding study on Wistar rats, the transgenic KMD1 rice expressing Cry1Ab protein was compared to its non-transgenic parental wild type, Xiushui 11. The KMD1 rice contained 15mg Bt toxin/kg and based on the average feed consumption the daily intake was 0.54mg Bt toxin/kg body weight. No adverse effects on animal behaviour or weight gain were observed during the study. Blood samples collected one week prior to sacrifice were analyzed and compared for standard haematological and biochemical parameters. A few parameters were significantly different, but all within the normal reference intervals for rats of this breed and age and not in relation to any other findings, thus not considered treatment related. Upon sacrifice a large number of organs were weighed, macroscopic and histopathological examinations were performed with only minor changes to report. The aim of the study was to use a known animal model in performance of safety assessment of a GM crop, in this case KMD1 rice. The results show no adverse or toxic effects of KMD1 rice when tested in the design used in this 90-day study. Nevertheless the experiences from this study lead to the overall conclusion that safety assessment for unintended effects of a GM crop cannot be done without additional test group(s).     

微生物发酵法生产利巴韦林的初步研究

以枯草芽孢杆菌（Bacillus subtilis）TM903（Ade^- ＋8-AG^t＋MSO^t）为生产菌株，考察了前体物1，2，4-三唑-3-甲酰胺（TCA）、MnSO4、温度、pH及培养基装量对发酵法生产利巴韦林的影响，确定了其摇瓶发酵工艺：培养至24h时添加10g／LTCA及10mg／LMnSO4，24h后每6h升温2℃，采用磷酸缓冲液调节pH，在500ml挡板三角瓶中培养基装液量为25ml。该工艺条件下经60h发酵，利巴韦林的摇瓶发酵水平达5．11g／L。     

卡介菌多糖核酸对尖锐湿疣病人外周血CD8~+T细胞细胞因子表达影响

目的:研究卡介菌多糖核酸(BCG-PSN)对尖锐湿疣(CA)病人外周血CD8q+T细胞内白细胞介素类(IL-2、IL-4、IL-12)和干扰素γ(IFN-γ)表达的影响。方法:60例CA病人分为2组,均采用CO_2激光去除疣体。BCG-PSN组40例,激光术后开始加用BCG-PSN 1.0mg,im,qod,连续18次,36d为一个疗程。病例对照组20例,激光术后未再用任何药物治疗。另设正常对照组20例。采用流式细胞仪测定3组外周血CD8+T细胞IL-2、IL-4、IL-12、IFN-γ的水平。结果:治疗前CA病人外周血IL-2、IL-12和IFN-γ阳性CD8+T细胞百分率显著低于正常对照组,Tcl/Tc2亦显著低于正常对照组(P<0.01)。治疗后BCG-PSN组外周血IL-2、IL-12和IFN-γ阳性CD8+T细胞百分率显著升高(P<0.01),病例对照组无显著变化(P>0.05),2组变化差异非常显著(P<0.01)。BCG-PSN组治愈率82％,显著高于病例对照组(45％,P<0.01);复发率18％,低于病例对照组(55％,P<0.01)。结论:CA病人存在Tcl/Tc2失调,BCG-PSN可通过上调CA病人外周血CD8+T细胞IL-2、IL-12、IFN-γ的表达,纠正机体细胞因子失调而提高CA病人临床疗效、减少其复发。     

海洋细菌Bacillus sp.中环二肽类代谢产物的研究

目的研究海洋细菌Bacillus sp.发酵液的正丁醇萃取物中的化学成分。方法采用硅胶柱色谱、凝胶柱色谱、高效液相色谱等分离手段从海洋细菌Bacillussp.发酵液的正丁醇萃取物中分离得到8个化合物,利用理化性质和波谱分析鉴定它们的结构。结果与结论分离得到8个已知的环二肽类化合物,分别鉴定为3-苄基哌嗪-2,5-二酮(1)、3-甲基哌嗪-2,5-二酮(2)、3-异丁基哌嗪-2,5-二酮(3)、3-异丁基-6-甲基哌嗪-2,5-二酮(4)、吡咯并哌嗪-1,4-二酮(5)、(3S,8aS)-3-甲基-吡咯并哌嗪-1,4-二酮(6)、3-异丁基-吡咯并哌嗪-1,4-二酮(7)、3-异丁基-8-羟基-吡咯并哌嗪-1,4-二酮(8)。8个化合物均为首次从该细菌代谢物中分离得到。     

Edible safety requirements and assessment standards for agricultural genetically modified organisms.

This paper describes the background, principles, concepts and methods of framing the technical regulation for edible safety requirement and assessment of agricultural genetically modified organisms (agri-GMOs) for Shenzhen Special Economic Zone in the People's Republic of China. It provides a set of systematic criteria for edible safety requirements and the assessment process for agri-GMOs. First, focusing on the degree of risk and impact of different agri-GMOs, we developed hazard grades for toxicity, allergenicity, anti-nutrition effects, and unintended effects and standards for the impact type of genetic manipulation. Second, for assessing edible safety, we developed indexes and standards for different hazard grades of recipient organisms, for the influence of types of genetic manipulation and hazard grades of agri-GMOs. To evaluate the applicability of these criteria and their congruency with other safety assessment systems for GMOs applied by related organizations all over the world, we selected some agri-GMOs (soybean, maize, potato, capsicum and yeast) as cases to put through our new assessment system, and compared our results with the previous assessments. It turned out that the result of each of the cases was congruent with the original assessment.     

一株具有抗稻瘟霉活性的海洋细菌32-11-2-2的筛选和分离鉴定

目的:从中国东海微生物样品中分离筛选活性细菌菌株,并对活性菌株32-11-2-2进行鉴定.方法:采用无限稀释和平板划线法分离微生物样品,利用稻瘟霉模型进行活性筛选;并对其中的活性菌株32-11-2-2的形态、培养特征以及生理生化特征进行研究,使用16S rDNA序列分析的方法对菌株32-11-2-2进行了分类鉴定.结果:分离得到421株细菌,包括活性菌株54株,其中活性菌株32-11-2-2是一株弱嗜盐菌,具有陆地芽孢杆菌的形态特点,能产生胞外淀粉酶、过氧化氢酶,液化明胶、石蕊牛奶反应阳性,16S rDNA序列分析该菌株与枯草芽孢杆菌(Bacillus subtilis)同源性达到98%以上.结论:活性菌株32-11-2-2是一株适应了海洋环境的枯草芽孢杆菌.