Molecular Characterisation of Very Virulent Infectious Bursal Disease Virus and the Effects of Its Viral Proteins on Cancer Cell Lines

Infectious bursa1 disease virus (IBDV) is an immunosuppressive virus causing bursa lesions and atrophy. There are several strains of IBDVs, namely classical, variant, attenuated and very virulent strains. Viral proteins, VP2 and VP5, from attenuated strain had been shown to induce apoptosis in chicken embryo fibroblast (CEF) and African green monkey cell line (BSC40). The objective of this study is to characterise local isolate IBDV namely, UPMOIIIO and to determine the apoptotic effect of VP2 and VP5 proteins in neoplastic and normal cell lines. UPMOIIIO was successfully characterised as vvlBDV, based on the molecular methods. The VP2 sequence of UPMOIIIO isolate had amino acid substitutions at Ala[222], [lle]256, [He1294 and [Ser]299, similar to other reported vvlBDV. This isolate did not have Lys[249] and Ser[254] amino acid residues which had been reported to be present in variant strains. The deduced amino acids of VP2 showed that the two hydrophilic regions and the serine-rich heptapeptide region were conserved in UPMOll10. Similar finding was reported for other vvlBDVs such as UK661, HK46 and OKYM. The VP2 nucleotides sequence of UPMO1110 could be cut by restriction enzymes Taql, Accl, Styl, Spel but not by Sacl. UPMO1/10 showed highest homologous similarity in nucleotides and amino acids to the reported Malaysian vvlBDVs. Phylogenetic analysis based on the nucleotides sequence of the VP2 and VP5 revealed that UPMO1110 isolate was clustered with other very virulent strains but distanced with classical, variant and attenuated strains. Serial samples of Chang, HepG2 and MCF-7 cells transfected with VP2 or VP5 at the interval of 6, 12, 18 and 24 hours were examined, respectively, in order to identify the onset of apoptosis. VP2 and VP5 of UPMO1110 were successfully expressed in Chang, HepG2 and MCF-7 cells. VP2 or VP5 alone was capable of triggering apoptotic response in Chang, HepG2 and MCF-7 cells. The onset of apoptosis started at 6 hours and preceded to necrosis at 24 hours. Both Chang and HepG2 cells were significantly susceptible to either VP5 or VP2 as compared to MCF-7 cells. VP5 induced significantly prominent apoptosis in HepG2 as compared to Chang cells at 6 hours, followed by maintaining similar level of apoptosis in both cells from 12 to 24 hours. However, VP2 induced higher level of cell death significantly in Chang cells as compared to HepG2 cells with exception at 6 hours. VP5 may be possible to be taken into consideration as candidate for liver cancer therapy as compared to VP2

Investigation of a Potential Zoonotic Transmission of Orthoreovirus Associated with Acute Influenza-Like Illness in an Adult Patient

Bats are increasingly being recognized as important reservoir hosts for a large number of viruses, some of them can be highly virulent when they infect human and livestock animals. Among the new bat zoonotic viruses discovered in recent years, several reoviruses (respiratory enteric orphan viruses) were found to be able to cause acute respiratory infections in humans, which included Melaka and Kampar viruses discovered in Malaysia, all of them belong to the genus Orthoreovirus, family Reoviridae. In this report, we describe the isolation of a highly related virus from an adult patient who suffered acute respiratory illness in Malaysia. Although there was no direct evidence of bat origin, epidemiological study indicated the potential exposure of the patient to bats before the onset of disease. The current study further demonstrates that spillover events of different strains of related orthoreoviruses from bats to humans are occurring on a regular basis, which calls for more intensive and systematic surveillances to fully assess the true public health impact of these newly discovered bat-borne zoonotic reoviruses

Screening of Streptococcus suis in swine workers of selected states in Peninsular Malaysia

Background and Aim: Streptococcus suis is a zoonotic pathogen that is highly associated with contact between live pigs and raw pig material. In view of the recent reports of human infections in Malaysia, epidemiological data on the status of S. suis in the human population, especially among people working closely with pigs and/or raw pork, should be provided. The aim of this study was to detect S. suis among individuals working in the swine industry in several major pig production areas in Peninsular Malaysia. Materials and Methods: Demographic information, exposure determinants, and oral swabs were collected from swine personnel, including farmers, butchers, and veterinarians. Oral swabs were subjected to bacterial isolation and conventional polymerase chain reaction (PCR) assays for S. suis detection. Results: The study included 40 participants working in the swine industry, with a predominant representation of males (62.5%) and Malaysian Chinese individuals (60.0%) who consumed pork (92.5%). Notably, none of the participants reported consuming raw or partially cooked pork. In spite of their occupational exposure risk, none of the oral swabs showed positive results for S. suis infection. Conclusion: To the best of our knowledge, this is the first report and detection study of S. suis using oral swabs obtained from swine personnel in Peninsular Malaysia

Screening of anti-dengue activity in methanolic extracts of medicinal plants

<p>Abstract</p> <p>Background</p> <p>Dengue fever regardless of its serotypes has been the most prevalent arthropod-borne viral diseases among the world population. The development of a dengue vaccine is complicated by the antibody-dependent enhancement effect. Thus, the development of a plant-based antiviral preparation promises a more potential alternative in combating dengue disease.</p> <p>Methods</p> <p>Present studies investigated the antiviral effects of standardised methanolic extracts of <it>Andrographis paniculata, Citrus limon, Cymbopogon citratus, Momordica charantia, Ocimum sanctum </it>and <it>Pelargonium citrosum </it>on dengue virus serotype 1 (DENV-1).</p> <p>Results</p> <p><it>O. sanctum </it>contained 88.6% of total flavonoids content, an amount that was the highest among all the six plants tested while the least was detected in <it>M. charantia</it>. In this study, the maximum non-toxic dose (MNTD) of the six medicinal plants was determined by testing the methanolic extracts against Vero E6 cells <it>in vitro</it>. Studies also determined that the MNTD of methanolic extract was in the decreasing order of <it>M. charantia </it>><it>C. limon </it>><it>P. citrosum, O. sanctum </it>><it>A. paniculata </it>><it>C. citratus</it>. Antiviral assay based on cytopathic effects (CPE) denoted by degree of inhibition upon treating DENV1-infected Vero E6 cells with MNTD of six medicinal plants showed that <it>A. paniculata </it>has the most antiviral inhibitory effects followed by <it>M. charantia</it>. These results were further verified with an <it>in vitro </it>inhibition assay using MTT, in which 113.0% and 98.0% of cell viability were recorded as opposed to 44.6% in DENV-1 infected cells. Although methanolic extracts of <it>O. sanctum </it>and <it>C. citratus </it>showed slight inhibition effect based on CPE, a significant inhibition was not reflected in MTT assay. Methanolic extracts of <it>C. limon </it>and <it>P. citrosum </it>did not prevent cytopathic effects or cell death from DENV-1.</p> <p>Conclusions</p> <p>The methanol extracts of <it>A. paniculata </it>and <it>M. charantia </it>possess the ability of inhibiting the activity of DENV-1 in <it>in vitro </it>assays. Both of these plants are worth to be further investigated and might be advantageous as an alternative for dengue treatment.</p

Identification and Characterization of a New Orthoreovirus from Patients with Acute Respiratory Infections

First discovered in the early 1950s, reoviruses (respiratory enteric orphan viruses) were not associated with any known disease, and hence named orphan viruses. Recently, our group reported the isolation of the Melaka virus from a patient with acute respiratory disease and provided data suggesting that this new orthoreovirus is capable of human-to-human transmission and is probably of bat origin. Here we report yet another Melaka-like reovirus (named Kampar virus) isolated from the throat swab of a 54 year old male patient in Kampar, Perak, Malaysia who was suffering from high fever, acute respiratory disease and vomiting at the time of virus isolation. Serological studies indicated that Kampar virus was transmitted from the index case to at least one other individual and caused respiratory disease in the contact case. Sequence analysis of the four small class genome segments indicated that Kampar and Melaka viruses are closely related. This was confirmed by virus neutralization assay, showing an effective two-way cross neutralization, i.e., the serum against one virus was able to neutralize the other. Although the exact origin of Kampar virus is unknown, epidemiological tracing revealed that the house of the index case is surrounded by fruit trees frequently visited by fruit bats. There is a high probability that Kampar virus originated from bats and was transmitted to humans via bat droppings or contaminated fruits. The discovery of Kampar virus highlights the increasing trend of emergence of bat zoonotic viruses and the need to expand our understanding of bats as a source of many unknown viruses

Reading Your Counterpart: The Benefit of Emotion Recognition Accuracy for Effectiveness in Negotiation

10.1007/s10919-007-0033-7Journal of Nonverbal Behavior314205-22