Blood-Brain Barrier Permeability of Asiaticoside,Madecassoside and Asiatic Acid in Porcine Brain Endothelial Cell Model

Neurotherapeutic potentials of Centella asiatica and its reputation to boost memory, prevent cognitive deficits and improve brain functions are widely acknowledged. The plant's bioactive compounds, i.e. asiaticoside, madecassoside and asiatic acid were reported to have central nervous system (CNS) actions, particularly in protecting the brain against neurodegenerative disorders. Hence, it is important for these compounds to cross the blood-brain barrier (BBB) to be clinically effective therapeutics. This study aimed to explore the capability of asiaticoside, madecassoside and asiatic acid to cross the BBB using in vitro BBB model from primary porcine brain endothelial cells (PBECs). Our findings showed that asiaticoside, madecassoside and asiatic acid are highly BBB permeable with apparent permeability (P_app) of 70.61 ± 6.60, 53.31 ± 12.55 and 50.94 ± 10.91 × 10^?6 cm/s respectively. No evidence of cytotoxicity and tight junction disruption of the PBECs were observed in the presence of these compounds. Asiatic acid showed cytoprotective effect towards the PBECs against oxidative stress. This study reported for the first time that Centella asiatica compounds demonstrated high capability to cross the BBB, comparable to central nervous system drugs, and therefore warrant further development as therapeutics for the treatment of neurodegenerative diseases.     

Development of an Enzyme-Linked Immunosorbent Assay Using a Recombinant LigA Fragment Comprising Repeat Domains 4 to 7.5 as an Antigen for Diagnosis of Equine Leptospirosis

Leptospira immunoglobulin (Ig)-like (Lig) proteins are a novel family of surface-associated proteins in which the N-terminal 630 amino acids are conserved. In this study, we truncated the LigA conserved region into 7 fragments comprising the 1st to 3rd (LigACon1-3), 4th to 7.5th (LigACon4-7.5), 4th (LigACon4), 4.5th to 5.5th (LigACon4.5–5.5), 5.5th to 6.5th (LigACon5.5–6.5), 4th to 5th (LigACon4-5), and 6th to 7.5th (LigACon6-7.5) repeat domains. All 7 recombinant Lig proteins were screened using a slot-shaped dot blot assay for the diagnosis of equine leptospirosis. Our results showed that LigACon4-7.5 is the best candidate diagnostic antigen in a slot-shaped dot blot assay. LigACon4-7.5 was further evaluated as an indirect enzyme-linked immunosorbent assay (ELISA) antigen for the detection of Leptospira antibodies in equine sera. This assay was evaluated with equine sera (n = 60) that were microscopic agglutination test (MAT) negative and sera (n = 220) that were MAT positive to the 5 serovars that most commonly cause equine leptospirosis. The indirect ELISA results showed that at a single serum dilution of 1:250, the sensitivity and specificity of ELISA were 80.0% and 87.2%, respectively, compared to those of MAT. In conclusion, an indirect ELISA was developed utilizing a recombinant LigA fragment comprising the 4th to 7.5th repeat domain (LigACon4-7.5) as a diagnostic antigen for equine leptospirosis. This ELISA was found to be sensitive and specific, and it yielded results that concurred with those of the standard MAT.     

Evaluation of inhibitory potential of some selective methanolic plants extracts on biological characteristics of Acanthamoeba castellanii using human corneal epithelial cells in vitro

Acanthamoeba is an opportunistic protozoan pathogen and known to be one of the most ubiquitous organisms, play a vital role in ecosystem, and recognized to cause blinding keratitis and rare but fatal granulomatous encephalitis involving the central nervous system with a very poor prognosis. This is due to limited availability of effective anti-Acanthamoeba drugs. The objective of the present study was to determine the efficacy of methanolic plants crude extracts on the viability and biological properties of Acanthamoeba castellanii (T4 genotype) and its cytotoxic effects on human corneal epithelial cells (HCEC). Using HCEC, it was observed that Acanthamoeba exhibited binding (>90 %) and cytotoxicity (>80 %) to host cells. However, plant crude extracts remarkably inhibited more than 70 and 60 % of Acanthamoeba binding and cytotoxicity to HCEC, respectively. It was further established that crude extracts (ranging from 0.1 to 1.5 mg/ml) exhibited amoebicidal effects, i.e., >50 % of trophozoites were killed/reduced at maximum dose (1.5 mg/ml) within 1 h incubation. However, the residual subpopulation remained static over longer incubations. Furthermore, growth assay demonstrated crude extracts inhibited >50 % Acanthamoeba numbers up to 7 days. Our results confirmed that plant crude extracts has inhibitory effects on Acanthamoeba growth and viability. Overall, these findings revealed that tested plant extracts is inhibitory to Acanthamoeba properties associated with pathogenesis. To the best of our knowledge, our findings demonstrated for the first time that selected methanol plant crude extracts exhibits inhibitory effects on biological properties of Acanthamoeba without any toxic effects on HCEC cells in vitro.     

Traumatic lumbar visceral herniation in a young woman

INTRODUCTIONLumbar herniation is uncommon, with traumatic etiology being rare. Traumatic lumbar hernias are usually caused by seatbelt injury in motor vehicle accidents. It is exceedingly uncommon to see lumbar hernias in an unrestrained passenger of a motor vehicle accident.PRESENTATION OF CASEWe present a case of a traumatic inferior lumbar hernia in a young woman who was an unrestrained driver of a vehicle involved in a high-speed collision, with multiple rollover and ejection. CT scans of the abdomen and pelvis suggested soft tissue injury involving muscles in the left lower posterior flank with traumatic herniation of the colon and small bowel. Emergent midline abdominal laparotomy confirmed herniation in the left lower quadrant. After abdominal closure, in the prone position, an extensive laceration over the left flank also confirmed herniation. Due to its dirty nature, the wound was irrigated, lavaged and covered with wound vacuum-assisted closure placement. The decision was made in favor of delayed elective hernia repair.DISCUSSIONLumbar hernias are usually caused by sudden force to the abdomen, leading to increased intra-abdominal pressure. This pressure combined with areas of weakness in the superior and/or inferior triangle lead to herniation. Uncommonly, the contents of lumbar hernias can strangulate or incarcerate leading to bowel obstruction. This can often be prevented by detection with CT and laparotomy.CONCLUSIONLumbar herniation of traumatic etiology is rare. Early detection with CT and/or exploratory laparotomy is important to avoid increases in size of the defect and bowel strangulation and incarceration.     

助视器对年龄相关性黄斑变性患者视功能恢复和阅读速度的影响

目的:评估助视器对年龄相关性黄斑变性(AMD)患者视功能恢复和阅读速度的影响。方法:对低视力门诊44名AMD患者进行评估。收集并分析使用助视器前和使用助视器时裸眼视力、最佳矫正视力、屈光状态、助视器类型(LVAs)以及阅读速度的相关数据。结果:共44例AMD患者,平均年龄为73±10.8岁,男性36例(82%)。其中,29例(67%)受试者视力较好的眼远视力(DVA)为1.0~1.6 LogMAR,而36例(82%)受试者近视力(NVA)小于3.2 m。使用助视器时平均DVA提高0.67±0.27 LogMAR(P=0.000)。使用LVAs时,42例患者NVA达到1 m甚至更佳,仅有2例(4.5%)没有使用LVAs的患者NVA为1 m。在使用一定时间的助视器后,有阅读能力的患者平均阅读速度从每分钟2.9±4.78字提高到每分钟71.31±29.96字(P<0.001)。15名受试者远距离视觉使用单目望远镜,而戴高倍单目镜片眼镜作为最常用的阅读辅助工具。结论:LVAs对AMD患者的视功能恢复和阅读能力的提高有一定的作用。对于AMD患者而言,接受低视力护理服务是有必要的。     

Guidelines and considerations for conducting experiments using tissue microarrays

Tissue microarrays (TMAs) represent a powerful method for undertaking large‐scale tissue‐based biomarker studies. While TMAs offer several advantages, there are a number of issues specific to their use which need to be considered when employing this method. Given the investment in TMA‐based research, guidance on design and execution of experiments will be of benefit and should help researchers new to TMA‐based studies to avoid known pitfalls. Furthermore, a consensus on quality standards for TMA‐based experiments should improve the robustness and reproducibility of studies, thereby increasing the likelihood of identifying clinically useful biomarkers. In order to address these issues, the National Cancer Research Institute Biomarker and Imaging Clinical Studies Group organized a 1‐day TMA workshop held in Nottingham in May 2012. The document herein summarizes the conclusions from the workshop. It includes guidance and considerations on all aspects of TMA‐based research, including the pre‐analytical stages of experimental design, the analytical stages of data acquisition, and the postanalytical stages of data analysis. A checklist is presented which can be used both for planning a TMA experiment and interpreting the results of such an experiment. For studies of cancer biomarkers, this checklist could be used as a supplement to the REMARK guidelines.     

A novel audio forensic data-set for digital multimedia forensics

Today, digital multimedia (audio, video, images) is a common evidential source in litigation and criminal justice proceedings, and, not surprisingly, multimedia forensics is an active research area. One particular challenge faced by multimedia forensic researchers is the lack of a comprehensive and publicly available data-set for evaluating existing and new algorithms. This paper presents a digital audio forensic data-set, designed to facilitate evaluation of audio forensic algorithms (e.g. microphone identification, acoustic environment identification and splice detection). This paper also briefly describes the data-collection settings, microphones, speakers, languages and notations used. Existing tamper-detection techniques rely on artefacts due to recording devices, codec and/or the acoustic environment in the audio in question. Experimental results show that the selected approaches achieved promising results.