Inhibition of Curcumin-Treated Herpes Simplex Virus 1 and 2 in Vero Cells

The purpose of this study was to investigate the effect of curcumin-treated Herpes simplex virus-1 (HSV-1) and Herpes simplex virus-2 (HSV-2) virions in cultured Vero cells. Previous studies have indicated that curcumin, a polyphenol extracted from the plant Curcuma longa, has demonstrated antiviral properties against a variety of viruses. After establishing the maximum non-cytotoxic concentrations of curcumin on Vero cells, HSV-1 and HSV-2 virions were treated with varying concentrations of curcumin. The effect on infectivity was determined by antiviral assays, using WST-1, plaque assays, adsorption and penetration assays. Treating HSV-1 and HSV-2 viruses with curcumin, at a concentration of 30 μM, reduces the production of infectious HSV-1 and HSV-2 virions in cultured Vero cells by interfering with the adsorption process. These results support the potential of curcumin to be used as a therapeutic agent to reduce the transmission of HSV-1 and HSV-2.

Using the SELDI ProteinChip System to Detect Changes in Protein Expression in Vero Cells after Infection

人的疱疹单一的病毒(HSV-1 ) 1 引起美容，眼睛，并且 encephalitic 疾病并且与潜伏的感染和癌症被联系。这里，我们开发了由使用 SELDI 蛋白质薄片在在 vitro 有教养的 Vero 房间检测蛋白质表示的变化在蛋白质水平学习 HSV-1 感染的致病的一个工具。在有为 12， 24 或 48 h 的 HSV-1 和文化的感染以后，房间被收获并且 lysed。IMAC3 数组被用于 SELDI-TOF-MS 在感染前后检测 proteomic 差别。薄片检测了一系列差别表示蛋白质山峰。有趣地，在 16 912 Da 的山峰和 17 581 Da 与 ISG15 的分子的团精确相应，它可以在感染的过程期间参予抗病毒的活动。因此，我们获得了的结果能用作一个基础学习在病毒和它的主人之间的 HSV-1 和相互作用的致病。另外，他们能为 HSV-1 感染的治疗在新治疗学的目标的发现帮助。关键词 SELDI 蛋白质薄片 - Vero 房间 - HSV-1 - 蛋白质表达式 CLC 数字 R373 基础项目：中国(30540075 ) 的国家自然科学基础；

Pilot-Scale Production of Lyophilized Inactivated Rabies Vaccine Candidate in Vero Cells under Fully Animal Component-Free Conditions Using Microcarrier Technology and Laboratory Animal Trials

The upstream process was carried out in an animal component-free medium on Cytodex 1 microcarriers. Recombinant trypsin is a non-animal derived protease used as an alternative to animal-derived trypsin. To inactivate recombinant trypsin, a soybean trypsin inhibitor (STI) should be added to the medium. A protocol was first tested in T-flasks and then passaged to 500 mL and 3 L spinner flasks. Cell detachment was completed in 10 - 12 min, and 0.4 g/L STI was added to a 3L spinner, and cells were transferred into a 30 L stirred tank bioreactor. On day 5, the cell density had reached its maximum (around 1.8 × 106 cells/mL). At an MOI of 0.3 with serum-free medium conditions, cell infection yielded a maximal rabies virus titer of 1.82 × 107 FFU/mL at 5 days. All cell culture conditions and virus growth kinetics in serum-free media were investigated. In conclusion, Vero cells were grown on Cytodex 1 with serum-free media and a high amount of rabies virus was obtained. A mouse challenge was used to determine the immune response to an inactivated rabies virus vaccine candidate. Also, we evaluated inactive rabies vaccine candidate safety, and immunogenicity in mice, sheep, horses, and cattle. We found that no horses, sheep, or cattle who were given vaccine IM at 3.2 IU/dose exhibited any clinical sign of disease and all developed high VNA titers (up to 10.03 IU/mL) by 3 - 4 WPI. After the accelerated stability studies, the lyophilized inactivated rabies vaccine candidate showed enough antigenic potency (2.6 IU/mL) in the mouse challenge test. Also, 18-month long-term stability studies showed enough immune response (1.93 IU/mL) on day 14. The activity of the vaccine candidate showed a good immune response and safety criteria that meet WHO requirements. This is the first pilot-scale mammalian cell-based viral rabies vaccine production study in Türkiye that used microcarriers.

Bacteroides fragilis Supernatant Extracts Enriched in Phenylacetic Acid Induce a Cytotoxic Effect in Mammalian Cells

Bacteroides species are nearly half of the fecal flora community and some are host symbionts crucial to host nutrition and systemic immunity. Among Bacteroides species B. fragilis strains are considered to be the opportunistic ones, being the most isolated anaerobic bacteria in clinical samples. Cell-free supernatants of 65 B. fragilis strains were assayed and they were capable of inducing vacuolating phenotype on Vero cells lineage. The supernatant of the Bacteroides fragilis ATCC 23745 strain was elicited to have the strongest vacuolating effect on Vero cells monolayers and peritoneal macrophages. Some drastic cell alterations were observed, such as a general disorganization of cytoplasm and chromatin condensation, evidencing cell death. By transmission electron microscopy it was confirmed that the vacuoles observed were, in fact, swollen mitochondria. An immunocytochemical assay, TUNEL, was used to confirm this hypothesis and showed that Vero cells and peritoneal macrophages were dying by apoptotic process after exposition of B. fragilis cell-free supernatant. Physical analysis of the apoptotic factor has revealed properties similar to short-chain fatty acids. After gas chromatography and mass spectrometry analysis, phenylacetic acid (PA) was characterized as the major compound present in the most purified active fraction. We believe that the PA is responsible for the pro-apoptotic effect elicited by the supernatant of B. fragilis cultures.

Agranulocytosis following injection of inactivated Japanese encephalitis vaccine(Vero cell):A case report

BACKGROUND Japanese encephalitis virus(JEV),a mosquito borne flavivirus,is the leading cause of viral encephalitis in Asia,in terms of frequency and severity.JEV infection is thought to confer lifelong immunity.With the near eradication of poliomyelitis,JEV is now the continent’s leading cause of childhood viral neurologic infection and disability.The most common clinical manifestation of JEV infection is acute encephalitis,and currently there is no specific antiviral therapy.Japanese Encephalitis Vaccine(JE-VC)is an effective prevention measure,including JE-VC,Live(JE-MB),and Inactivated JE-VC.CASE SUMMARY A 9-mo-old girl received injection of Inactivated JE-VC(Vero cell)(Liaoning Chengda,batch number 201611B17)on August 31,2017.On that night,she developed a fever with the body temperature up to 38.5°C,for which Ibuprofen Suspension Drops 1.25 mL was given as antipyretic treatment.On September 1,the patient developed apocleisis,and her parents noticed herpes in her oral cavity.The patient was sent to our hospital on September 3.Physical examination led to a diagnosis of herpetic stomatitis,for which Stomatitis Spray 1 puff,tid,Kangfuxin Liquid 2 mL,tid,and vitamin B20.5 tablet,tid,were prescribed.Routine blood tests for low fever on September 6,2017 revealed an absolute neutrophil count(ANC)of 0.62×109/L,hemoglobin(Hb)of 109 g/L,and platelet count(PLT)of 308×10^(12)/L,and the tests were monitored regularly thereafter.The patient was followed until July 26,2020,when routine blood tests revealed ANC 1.72×109/L,Hb 138 g/L,and PLT 309×1012/L,indicating that the neutropenia count had normalized.CONCLUSION This report attempts to bring to clinical attention that Inactivated JE-VC(Vero cell)might cause prolonged granulocytopenia or even agranulocytosis.

Evaluation of the Antiviral Activity of Propolis from Native Bees (<i>Plebeia frontalis</i>) against Canine Distemper Virus

Propolis is a natural substance made from resins collected from trees and plants, and which bees combine with pollen, wax, and their own enzymes. It has a complex chemical composition that varies with the harvest season, vegetation type, bee species, and geographical region. Thanks to its components, it has valuable biological properties such as antifungal, antibacterial, anticancer, antiviral, and immunomodulatory activity. For this study, a sample of propolis harvested in April 2019 was used, which came from a bee native to Mexico (Plebeia frontalis) in whose geographical environment there are seven other native species. Canine distemper virus is an RNA virus that causes a systemic infection with high fatality rates in guests without protective immunity. In this work, the antiviral effect of Plebeia frontalis propolis on canine distemper virus was tested, administering it one hour before and simultaneously to infection. The antiviral effect was evaluated by determining cellular viability with the MTT assay. The results obtained show that this propolis has a statistically significant antiviral effect on both treatments, although it is slightly better when applied one hour before viral infection, so we can recommend it as an antiviral treatment in both domestic animals and human beings. There are currently few studies of the antiviral effect of propolis, this being the first study of a melliponium propolis in veterinary medicine.

Ketamine effect on intracellular and mitochondrial calcium mobilization

The suppressive effects of ketamine on intracellular calcium has been reported in a variety of cells although the mechanisms involved are not well understood.The aim of this work was to evaluate the ketamine effect on the mitochondrial Ca^(2+)accumulation and the cellular Ca^(2+)mobilization using FLUO4-AM and flow cytometry.The results showed that mitochondria from ketamine injected animals presented a lower ability to retain calcium at concentrations higher than 20μM,as compared with controls(saline injected animals).In ad-dition,ketamine showed a significant decreased KCl-induced intracellular calcium concentration.KCl increased calcium influx through cellular depolarization.According to the data presented herein,ketamine presents a clear inhibitory effect on cytosolic Ca2+transport mechanisms,independently from their action on the calcium channel associated NMDA receptor.

Antiviral effect of polyphenol rich plant extracts on herpes simplex virus type 1

Evaluation of the anti-herpetic activity of 25 Egyptian plants extracts was investigated in vitro in this study.The antiviral activity against Herpes Simplex Virus type 1(HSV-1)was done on Vero cell lines by cell viability.Only two plants extracts;namely Euphorbia coopire(Euphorbiaceae)and Morus alba(Moraceae)showed potent anti-herpetic activity and six other extracts showed moderate inhibition.In contrast,a bioassay monitored phytochemical exploration of these two plants led to the isolation of pure flavonoid compounds.The antiviral activity of the isolated compounds was also examined,among which seven pure compounds namely;7-galloyl catechin,gallic acid,kaempferol 3-O-β-(6″-O-galloyl)-glucopyranoside,quercetin 3-O-β-(6″-O-galloyl)-glucopyranoside,curcumin,quercetin and kaempferol exhibited significant inhibition.

顽固性扁平疣患者接种新型冠状病毒灭活疫苗后皮损自行消退一例

患者,男,26岁。2年前无明显诱因面部、上肢出现多发扁平丘疹,曾多次就诊于我院,诊断为"扁平疣"给予维A酸类药物外用、CO;点阵激光治疗,效果欠佳。2021年5月13日患者行COVID-19-1(Vero Cell)(新型冠状病毒灭活疫苗-北京生物?,批号2021020088)首剂接种,1天后扁平疣皮损变红,自觉瘙痒明显,于2021年5月17日来我院就诊,未予药物治疗,嘱观察,2021年5月23日复诊时疣体枯萎脱落。