Induction of apoptosis on human hepatocarcinoma cell lines by an alkyl resorcinol isolated from Lithraea molleoides

AIM: To study the mechanism of cytotoxicity of a new active 5-alkyl resorcinol [1, 3-dihydroxy-5- (tridec-4’, 7’ -dienyl) benzene] isolated from Lithraea molleoides leaves on liver tumor cells. METHODS: Human hepatocarcinoma cell lines (HepG2 and Hep3B) in culture were treated with inhibitory concentrations, 50% of the compound, for 24 h. The induction of apoptosis was detected in treated cells by analysis of DNA fragmentation, DNA content, and acridine orange and propidium iodide staining. RESULTS: After 24 h of 5-alkyl resorcinol treatment, both cell lines showed: (1) the typical morphological alterations of apoptosis; (2) DNA fragmentation, detected by laddering and appearance of a subG0 population by flow cytometry; and (3) condensed and fragmented nuclei by acridine orange-propidium iodide staining. CONCLUSION: Based on the results, this compound exerts its cytotoxic effect in both hepatocellular cell lines through apoptotic cell death. For Hep3B, cells with mutated p53 and Fas, apoptosis would proceed by p53- or Fas-independent pathways.

Relaxin prevents the development of severe acute pancreatitis

瞄准：为了调查尖锐胰腺炎(AP ) 的严厉，被联系到白血球激活，煽动性的起来规定和联系到 ischemia-reperfusion 损害的微循环混乱的紧张。支持 inflammatory 调停人的 Microvascular 完整和抑制在 AP 的进化是关键因素的。松弛激素是被归因的像胰岛素的荷尔蒙经由氮的氧化物小径的 vasorelaxant 性质当作为糖皮质激素受体收缩筋表现时。方法：AP 被 bilio 胰腺的 duct-outlet-exclusion closed-duodenal-loops 模型导致。有松弛激素的处理在不同时间点被做。由由 mifepristone 的阻塞被考虑的 L 名字和糖皮质激素受体(GR ) 的氮的氧化物 synthase 抑制。AP 严厉被生物化学、组织病理学说的分析估计。结果：有松弛激素的处理减少了浆液淀粉酶，脂肪分解酵素， C 反应的蛋白质， IL-6， IL-10， hsp72， LDH 和 8-isoprostane 象一样胰腺并且肺 myeloperoxidase。腺泡和脂肪坏死，出血和 neutrophil 渗透也被减少。当 caspases 2-3-8 和 9 项活动被增加时， ATP 弄空和 ADP/ATP 比率被减少。L 名字和 mifepristone 减少了松弛激素的效率。结论：松弛激素结果在当保存微发行量并且在坏死上赞成 apoptosis 时，联合 GR 收缩筋的性质的 AP 的处理有益。

Acute hepatitis C in a chronically HIV-infected patient:Evolution of different viral genomic regions

AIM: To analyze the molecular evolution of different viral genomic regions of HCV in an acute HCV infected patient chronically infected with HIV through a 42-month follow-up.METHODS: Serum samples of a chronically HIV infected patient that seroconverted to anti HCV antibodies were sequenced, from the event of superinfection through a period of 17 months and in a late sample (42nd month). Hypervariable genomic regions of HIV (V3 loop of the gp120) and HCV (HVR-1 on the E2 glycoprotein gene) were studied. In order to analyze genomic regions involved in different biological functions and with the cellular immune response, HCV core and NS5A were also chosen to be sequenced. Amplification of the different regions was done by RT-PCR and directly sequenced. Confirmation of sequences was done on reamplified material. Nucleotide sequences of the different time points were aligned with CLUSTAL W 1.5, and the corresponding amino acid ones were deduced.RESULTS: Hypervariable genomic regions of both viruses (HVR1 and gp120 V3 loop) presented several nonsynonymous changes but, while in the gp120 V3 loop mutations were detected in the sample obtained right after HCV superinfection and maintained throughout, they occurred following a sequential and cumulative pattern in the HVR1. In the NS5A region of HCV, two amino acid changes were detected during the follow-up period, whereas the core region presented several amino acid replacements, once the HCV chronic infection had been established.CONCLUSION: During the HIV-HCV superinfection, each genomic region analyzed shows a different evolutionary pattem.Most of the nucleotide substitutions observed are nonsynonymous and clustered in previously described epitopes,thus suggesting an immune-driven evolutionary process.

Trypanosoma rangeli:growth in mammalian cells in vitro and action of a repositioned drug(17-AAG)and a natural extract(Artemisia sp.essential oil)

Trypanosoma rangeli and T.cruzi are both parasitic unicellular species that infect humans.Unlike T.cruzi,the causative agent of Chagas disease,T.rangeli is an infective and non-pathogenic parasite for humans,but pathogenic for vectors from the Rhodnius genus.Because both species can coexist in different hosts and overlap their infective cycles but very little is known about the infection of T.rangeli in mammalian cells,we decided to characterize both the development of this parasite in cell culture and the effect of therapeutic agents with potential trypanocidal action on it.We found that T.rangeli exhibits a cycle of infection in Vero cells similar to that for T.cruzi and that the repurposed drug,17-AAG,and the natural extract Artemisia sp.essential oil produce a toxic effect on epimastigotes showing a trypanocidal action from the fifth day of culture.Both treatments also affected the infection of trypomastigotes and reduced the capacity of replication of amastigotes of T.rangeli.Since T.cruzi/T.rangeli coinfection cases have been reported,the finding of drugs with potential activity against both species could be significant in the future.Furthermore,studies of susceptibility of both species to drugs could also help to know the different mechanisms of pathogenicity in humans displayed by T.cruzi that are absent in T.rangeli.

Oxidative effects of the harmful algal blooms on primary organisms of the food web

Degraded water quality from nutrient pollution,physical,biological,and other chemical factors contributes to the development and persistence of many harmful algal blooms(HAB s).The complex dynamics of the HAB s is a challenge to marine ecosystems for the toxic effects reported.The consequences include fish,bird,and mammal mortality,respiratory or digestive tract problems,memory loss,seizures,lesions and skin irritation in many organisms.This review is intended to briefly summarize the recent reported information on harmful marine toxin deleterious effects over the primary organisms of the food web,namely algae,zooplankton and invertebrates.Special focus is made on oxidative stress status of cells and tissues.Even though in situ field research is less controlled than laboratory studies,in which the organisms are directly exposed to the toxins under consideration,both types of approaches are required to fully understand such a complex scenario.On top of that,the contribution of the increasing water temperatures in the sea,as a consequence of the global climate change,will be addressed as a topic for further studies,to evaluate the effect on regulating algal growth,species composition,trophic structure,metabolic stress and function of aquatic ecosystems.

Hepatitis C virus treatment failure:Clinical utility for testing resistance-associated substitutions

The hepatitis C virus has a high mutation capacity that leads to the emergence of resistance-associated substitutions(RAS).However,the consequence of resistance selection during new direct-acting antiviral drug(DAA)treatment is not necessarily the therapeutic failure.In fact,DAA treatment has shown a high rate(>95%)of sustained virological response even when high baseline RAS prevalence has been reported.In the context of RAS emergence and high rates of sustained viral response,the clinical relevance of variants harboring RAS is still controversial.Therefore,in order to summarize the data available in international guidelines,we have reviewed the clinical utility of testing RAS in the era of new pangenotypic DAA drugs.

Callogenesis and Antibacterial Activity of Balanites aegyptiaca

B. aegyptiaca, it is a species of economic and cultural importance in various countries, with diverse uses that include: medicinal, charcoal, pesticides and forage and in vitro callus production is important to have many applications in both basic and industrial research on this specie. For the induction of callus, B. aegyptica seed cotyledons were surface sterilized with 90% ethanol for 1 minute and cultivated in MS media supplemented with 2,4-D, BAP and NAA. Both the callus and seed were collected and dried in an oven at 40°C - 45°C. Cotyledon’s seed and callus were grounded into the powdered form using mortar and pestle and stored at room temperature for further use. Five grams (5 g) each of the powder were mixed with 50 ml of the solvents: methanol and n-hexane (1:10) w/v, agitated vigorously and kept on an orbital shaker at 150 rpm for 24 h, then filtered. The extracts of the plant sample were evaluated in agar dilution method which was used to determine the MIC and MBC of the extracts. The auxin NAA in low concentrations (0.5 mg/L) in the presence of a dose of 0.5 mg/L of the cytokinin BAP induced 100% callus formation. The 50 and 100 mg/ml methanolic extracts were more effective than the n-hexane extracts for both the gram-positive and gram-negative bacteria. By callus extracts under 100 and 50 mg/ml reveals that methanolic extracts of callus had the highest zone of inhibition. An effective protocol for callus induction has been developed that can use for germplasm conservation or for genetic engineering. Evidence from the present study revealed both extracts possess strong broad-spectrum antibacterial effect. Therefore, methanolic extract of seed kernel callus of B. aegyptiaca can be utilized as a new source of broad spectrum antibacterial drugs for effective control of bacteria related diseases.