■囊胚是一种常见的肠道寄生虫,具有全球分布。许多抗菌剂对它有效,然而,已经报道了副作用和耐药性。因此,正在进行的试验,以探索抗囊胚菌的替代品.蛋白酶是有吸引力的抗原生动物药物靶标,在囊胚病中有记录的作用。丝氨酸蛋白酶存在于丙型肝炎病毒和胚泡中。因为药物重新定位很时髦,simeprevir(SMV)的体外功效,一种抗肝炎丝氨酸蛋白酶抑制剂,在本研究中对胚泡进行了研究。
■收集患者的粪便样本,亚历山大,埃及。使用直接涂片筛选浓缩的粪便,三色,和改良的Ziehl-Neelsen染色以排除寄生虫共感染。培养阳性粪便分离株,分子分型用于评估三种SMV剂量(100,150和200μg/ml)在72小时(h)期间的功效,在最常见的亚型上,通过监测寄生虫的生长,生存能力,重新文化,也通过超微结构验证。随后在其他亚型上测试了最有效的剂量和持续时间。
■结果显示,在54.17%的检查样品中检测到胚泡。分子上,ST3占主导地位(62%),其次是ST1(8.6%)和ST2(3.4%)。SMV浓度的上升逐渐抑制生长,生存能力,和重新培养治疗的囊胚,与治疗性对照甲硝唑(MTZ)相比,差异无统计学意义。对ST3的最有效剂量和持续时间为150µg/ml,持续72小时。该剂量抑制了ST3,ST1和ST2的生长,百分比为95.19%,94.83%,94.74%,连续和生存力的百分比为98.30%,98.09%,和97.96%,先后。该剂量在再培养时消除了胚泡。超结构,SMV诱导的囊胚细胞膜破裂导致坏死死亡,与报告的由MTZ引起的凋亡死亡相比。总之,150µg/mlSMV持续72小时证明了其对ST1,ST2和ST3胚泡的功效,从而节省了发展中国家对预处理分子亚型的需求。
UNASSIGNED: Blastocystis is a common enteric parasite, having a worldwide distribution. Many antimicrobial agents are effective against it, yet side effects and drug resistance have been reported. Thus, ongoing trials are being conducted for exploring anti-Blastocystis alternatives. Proteases are attractive anti-protozoal drug targets, having documented roles in Blastocystis. Serine proteases are present in both hepatitis C virus and Blastocystis. Since drug repositioning is quite trendy, the in vitro efficacy of simeprevir (SMV), an anti-hepatitis serine protease inhibitor, against Blastocystis was investigated in the current study.
UNASSIGNED: Stool samples were collected from patients, Alexandria, Egypt. Concentrated stools were screened using direct smears, trichrome, and modified Ziehl-Neelsen stains to exclude parasitic co-infections. Positive stool isolates were cultivated, molecularly subtyped for assessing the efficacy of three SMV doses (100,150, and 200 μg/ml) along 72 hours (h), on the most common subtype, through monitoring parasite growth, viability, re-culture, and also via ultrastructure verification. The most efficient dose and duration were later tested on other subtypes.
UNASSIGNED: Results revealed that Blastocystis was detected in 54.17% of examined samples. Molecularly, ST3 predominated (62%), followed by ST1 (8.6%) and ST2 (3.4%). Ascending concentrations of SMV progressively inhibited growth, viability, and re-culture of treated Blastocystis, with a non-statistically significant difference when compared to the therapeutic control metronidazole (MTZ). The most efficient dose and duration against ST3 was 150 µg/ml for 72 h. This dose inhibited the growth of ST3, ST1, and ST2 with percentages of 95.19%, 94.83%, and 94.74%, successively and viability with percentages of 98.30%, 98.09%, and 97.96%, successively. This dose abolished Blastocystis upon re-culturing. Ultra-structurally, SMV induced rupture of Blastocystis cell membrane leading to necrotic death, versus the reported apoptotic death caused by MTZ. In conclusion, 150 µg/ml SMV for 72 h proved its efficacy against ST1, ST2, and ST3 Blastocystis, thus sparing the need for pre-treatment molecular subtyping in developing countries.