Municipal wastewater treatment plants as removal systems and environmental sources of human-virulent microsporidian spores

Municipal wastewater treatment plants play a vital role in reducing the microbial load of sewage before the end-products are discharged to surface waters (final effluent) or local environments (biosolids). This study was to investigate the presence of human-virulent microsporidian spores (Enterocytozoon bieneusi, Encephalitozoon intestinalis, and Encephalitozoon hellem) and enterococci during treatment processes at four Irish municipal secondary wastewater treatment plants (plants A–D). Microsporidian abundance was significantly related to seasonal increase in water temperature. Plant A had the least efficient removal of E. intestinalis spores (32%) in wastewater, with almost 100% removal at other plants both in April and July. Some negative removal efficiencies were obtained for E. bieneusi (at plants C and D, −100%) and for E. hellem (at plants A and D, −90% and −50%). In addition, a positive correlation was found between the levels of enterococci and E. bieneusi in July (r _s = 0.72, P < 0.05). In terms of the dewatered biosolids, a median concentration as high as 32,000 spores/Kg of E. hellem was observed at plant D in July. Plant C sewage sludge contained the lowest microsporidian loadings (E. bieneusi; 450 spores/L and 1,000 spores/L in April and July, respectively). This study highlights the seasonal variation in concentrations of microsporidian spores in the incoming sewage. Spores in final effluents and dewatered biosolids can be the source of human-virulent microsporidian contamination to the local environment. This emphasizes a considerably high public health risk when sewage-derived biosolids are spread during summer months. This study also suggested enterococci as a potential indicator of the presence of microsporidian spores in wastewater, especially for E. bieneusi.     

Mutagenicity and genotoxicity effects of Lignosus rhinocerotis mushroom mycelium

Ethnopharmacological relevance: Lignosus rhinocerotis mushroom is widely used as traditional medicine and as soup ingredient in Malaysia and Hong Kong. Its sclerotium is the part of edibility and is traditionally used for the treatment of fever, cough, asthma and cancer. In view of its safety profile, very little information is found in scientific literature.     

Factors affecting herpes simplex virus reactivation from the explanted mouse brain

The majority of encephalitis induced by herpes simplex virus type I (HSV-1) is due to viral reactivation from latency, but few studies have investigated the factors influencing viral reactivation in the brain due to the lack of a sensitive assay. We have established an ex vivo explant assay, which induced efficient viral reactivation in the dissociated mouse brain. Applying this assay, we investigated the infection of four HSV-1 strains with varying degrees of neurovirulence in three mouse strains with different levels of susceptibility to HSV-1 infection. We found that virulent HSV-1 strains and susceptible mouse strains exhibited prolonged viral growth during acute infection, increased latent viral genomes, and efficient explant reactivation in the brain stem. Collectively, both viral neurovirulence and host susceptibility positively correlate with HSV-1 reactivation from the explanted mouse brain.     

Beta-galactosidase deficiencies and novel GLB1 mutations in three Chinese patients with Morquio B disease or GM1 gangliosidosis

Background

This paper aims to report GLB1 activities and mutation analysis of three patients from the mainland of China, one with Morquio B disease and two with GM1 gangliosidosis.

Methods

GLB1 activity and GLB1 gene mutation were analyzed in the three patients who were clinically suspected of having Morquio B disease or GM1 gangliosidosis. Novel mutations were analyzed by aligning GLB1 homologs, 100 control chromosomes, and the PolyPhen-2 tool.

Results

The enzymatic activity of GLB1 was found to be 5.03, 4.20, and 4.50 nmol/h/mg in the three patients, respectively. Patient 1 was a compound heterozygote for p.[Arg148Cys] and p.[Tyr485Cys] mutations in the GLB1 gene. Patient 2 was a compound heterozygote for p.[Tyr270Phe] and p.[Leu337Pro] mutations. Patient 3 was a homozygote for p.[Asp448Val] mutation. Three mutations (p.[Tyr485Cys], p.[Tyr270Phe] and p.[Leu337Pro]) were novel variants and were predicted to damage GLB1 function.

Conclusions

The enzymatic activity and related gene analysis of ??-galactosidase should be performed in clinically suspected individuals to confirm diagnosis. The three novel mutations, p.[Tyr485Cys], p.[Tyr270Phe], and p.[Leu337Pro], are thought to be disease-causing mutations.     

Minocycline对血管性抑郁小鼠抑郁行为及神经递质的作用

目的 探讨minocycline抑制炎症反应对血管性抑郁小鼠行为及神经递质的影响。 方法 成年雄性CD1小鼠随机分为3组,每组各10只,实验组造模后立即腹腔注射minocycline每日1 次连续7 d（30 mg/kg）,对照组造模后给予同等剂量的生理盐水,假手术组除不阻断颈动脉供 血外,其余手术操作与实验组相同,术后同样给予腹腔注射相应剂量的生理盐水。术后第8天起 行悬尾实验（第8天）、旷场实验（第9天）检测抑郁行为,水迷宫定向航行实验（第10天）检测认 知能力。术后第11天处死取脑,分离海马匀浆,酶联免疫吸附测定（enzyme-linked immunosorbent assay,ELISA）试剂盒检测肿瘤坏死因子-α（tumor necrosis factor α,TNF-α）、白细胞介素-1β （interleukin-1β,IL-1β）及白细胞介素-6（interleukin-6,IL-6）含量,并通过高效液相色谱法（high performance liquid chromatography,HPLC）对抑郁相关的单胺类神经递质进行检测,包括5-羟色胺 （5-hydroxytryptamine,5-HT）、去甲肾上腺素（norepinephrine,NE）及多巴胺（dopamine,DA）。 结果 3组小鼠悬尾实验不动时间差异有显著性［实验组：（174.7 5±11.37）s,对照组： （194.32±14.32）s,假手术组：（169.62±19.27）s,F =6.59,P =0.005］;与对照组相比,实验组和假手 术组悬尾不动时间显著缩短;3组小鼠探洞次数、活动时间、活动路程差异显著（F =6.17,P =0.008; F =11.55,P＜0.001;F =13.47,P＜0.001）;与对照组相比,实验组与假手术组探洞次数明显增多［实 验组：（50.86±9.23）次,对照组：（35.73±11.96）次,假手术组：（48.14±10.16）次］,活动时 间与活动总路程均明显延长［实验组：（786.70±27.51）s,对照组：（738.88±36.00）s,假手术组： （807.90±33.16）s;实验组：（37 171.42±8493.40）mm,对照组：（28 992.91±5760.03）mm,假手术 组：（47 206.23±8219.84）mm］;水迷宫实验潜伏期差异有显著性［实验组：（87.38±13.36）s,对照 组：（88.50±19.88）s,假手术组：（44.38±19.76）s,F =16.09,P＜0.001］,与假手术组相比,实验组 和对照组潜伏期显著延长。炎症因子检测提示,3组海马TNF-α、IL-1β及IL-6差异具有显著性［实 验组：（141.10±24.36）pg/100 mg,对照组：（167.6±15.91）pg/100 mg,假手术组：（123.8±15.53） pg/100 mg,F =13.42,P＜0.001;实验组：（5.32±1.89）pg/mg,对照组：（10.31±2.83）pg/mg,假手术组： （4.50±2.07）pg/mg,F =18.69,P＜0.001;实验组：（20.01±3.62）pg/mg,对照组：（24.39±5.04）pg/mg, 假手术组：（18.40±3.78）pg/mg,F =5.49,P =0.010］;与对照组相比,实验组与假手术组3种炎性 细胞因子均显著降低。3组海马5-HT及DA含量差异有显著［实验组：（3.89±1.21）ng/ml,对照组： （3.13±1.44）ng/ml,假手术组：（5.01±1.68）ng/ml,F =4.17,P =0.026;实验组：（10.72±2.65）ng/ml, 对照组：（7.99±2.31）ng/ml,假手术组：（11.76±3.10）ng/ml,F =5.18,P =0.012］;与对照组相 比,实验组和假手术组DA含量显著增加,实验组5-HT含量差异无显著性,而假手术组5-HT含量增 多;3组NE含量差异无显著性［实验组：（3.97±1.35）ng/ml,对照组：（3.16±1.55）ng/ml,假手术组： （4.68±1.99）ng/ml,F =2.13,P =0.139］。 结论 Minocycline能够抑制血管性抑郁小鼠炎症因子的表达,抗炎症治疗可改善其抑郁行为,对认 知损害未观察到明显改善,相关的神经递质以DA的改变为最明显。     

血管性认知障碍小鼠脑内少突胶质细胞再生分化障碍特点研究

目的 观察血管性认知障碍小鼠模型中,缺血性炎性损伤对室管膜下区及海马齿状回少突胶质细胞 再生分化的影响,为血管性认知障碍的缺血性炎症机制提出新的损伤途径。 方法 成年雄性CD1小鼠随机分为模型组和假手术组,每组24只,模型组采用双侧颈动脉反复缺 血再灌注法制备血管性认知障碍小鼠模型。造模后4～6 d连续腹腔注射5 -溴脱氧尿嘧啶核苷 （bromodeoxyuridine,BrdU）（150 mg/kg）标记新生细胞,分别于术后14 d和28 d每组随机取一半小鼠脑 组织进行脑切片免疫组化、免疫荧光双标共聚焦检测,标记脑组织室管膜下区和海马区的少突胶质 细胞、星形胶质细胞及神经元,观察新生少突胶质细胞增殖及分化情况,并观察星形胶质细胞的增 生活化情况。 结果 造模后14 d和28 d室管膜下区新生细胞（BrdU阳性细胞）在模型组较假手术组明显增加（P均 ＜0.001),造模28 d模型组新生神经元（BrdU/NeuN阳性细胞）较假手术组显著增加（P＜0.001)。与假 手术组相比较,术后28 d模型组海马齿状回少突胶质细胞祖细胞显著增多（P＜0.001）;少突胶质细 胞前体细胞显著减少（P =0.006）。造模后28 d模型组海马齿状回新生星形胶质细胞（BrdU/GFAP阳性 细胞）较假手术组显著增加（P =0.015）。 结论 血管性认知障碍小鼠内源性新生细胞增殖区室管膜下区与海马齿状回区均存在新生细胞反 应性增生的情况。新生细胞区分化的主要细胞为星形胶质细胞,而少突胶质细胞分化障碍,可能是血 管性认知障碍患者影像学常见皮层下白质病变的重要原因。     

Regulatory analysis on the medical use of ephedrine-related products in Taiwan

To prevent ephedrine-related products from being misused to produce amphetamine and/or its analogs, there's a need for more effective and achievable regulatory mechanisms for the health, police, investigational, prosecution and judiciary authorities in Taiwan. This review was conducted to evaluate the international and Taiwan's regulatory policies and management of medical ephedrine-related products through the corresponding information collected from international and Taiwan government agency authorities. The combat of illegal drugs should involve both supply and demand sides to be successful. Health authorities in Taiwan do not have the investigational power to manage the forbidden transformation, abusing and manufacture of the illegal drugs from ephedrine-related products. Take the judicial interventions in the United States and in Japan as the examples, the organizational cooperation in Taiwan can be one of the main key strategies to combat against illegal drugs from ephedrine-related products. It is necessary to integrate the judicial, police and health agencies to prevent the production of illegal drugs from the ephedrine-related products in Taiwan. The efforts and regulatory control measures should be integrated to speed up the collaboration between different government authorities. It might be achieved through reorganization involving Taiwan Food and Drug Administration.