Identification of Phosphoprotien : Phosphoprotien and Phosphoprotien : Nucleocapsid Protein Interaction Domains of New Castle Disease Virus

Summary : The yeast two hybrid system has been used to identify domains of the Newcastle disease virus ( NDV ) phosphoprotien (P) involved in self –association and interaction with the nucleocapsid protein (NP). Deletion analysis was used to map the domain (s) of the P protein involved in P:P and P:NP interactions. The C- terminal 45 amino acids (residues 247-291) were shown to play a major role in both of interaction. Comparison of these finding with other report suggests that paramyxoviruses are different with respect to interaction domain(s) between these two essential viral proteins involved in genome replication

The matrix (M) protein of newcastle disease virus binds to human bax through its BH3 domain

The underlying mechanisms by which Newcastle disease virus (NDV) kills cancer cells are still unclear. Recent discoveries have shown that many viruses contain Bcl-2 homology-like domains which enabled their interaction with Bcl-2 family members, and thereby accounting for their virulence and pathogenicity. Alignment of the protein sequences of Malaysian strain of NDV, known as AF2240, with those from members of the human Bcl-2 family showed many similar regions; most notably we found that its matrix (AF2240-M) protein, large (AF2240-L) protein and fusion (AF2240-F) protein all contain BH3-like regions. In addition, there are BH1-like domains in these proteins, where AF2240-F and Mcl-1 share 55% identity within this region. To further investigate our hypothesis that the presence of the BH3-like domains in these proteins may convey cytotoxicity, AF2240-M and AF2240-F genes were cloned into pFLAG and pEGFP.N2 vectors and transfected into HeLa cells. The expression of these constructs promoted cell death. As shown by flow cytometry, AF2240-M protein with deleted BH3-like region showed five-fold decrease in apoptosis. Moreover, the construct containing the N-terminal of AF2240-M showed nearly the same cell death rate as to that of the full-length protein, strongly suggesting that the BH3-like domain within this protein participates in promoting cell death. Moreover, AF2240-M transfection promoted Bax redistribution to mitochondria. Therefore, to determine whether there is any direct interaction between NDV viral proteins with some members of the Bcl-2 family, various constructs were co-transfected into HeLa cells. Co-immunoprecipitation trials showed that the AF2240-M indeed directly interacted with Bax protein via its BH3-domain, as the mutant proteins failed to interact with Bax. AF2240-F failed to interact with any of the tested proteins, although Bcl-XL slowed down the rate of cell death caused by this construct by nearly five-fold. In a parallel experiment, the level of expression of endogenous Bax and Bcl-2 after infection of HeLa cells with NDV was assessed by qRT-PCR, but no statistically significant change was observed. Consequently, the Bax/Bcl-2 ratio at the mRNA level did not alter. Overall, our study has shed additional light into the mechanisms by which NDV induces apoptosis

Antioxidant research in Asia in the period from 2000-2008.

Problem statement: Plant antioxidants are natural plant products that inhibit the adverse effects of the Reactive Oxygen Species (ROS) produced in plants. Thus, antioxidants enable plants to survive. Depending on this principle, human beings started using some plants' parts to cure from diseases many centuries ago, even before the discovery of microbes or disclosing the etiology of many human diseases. Shed light on the importance of diet and natural products that have proved of great health impact on human traditionally and scientifically, disclosing the central role of natural antioxidants in this matter. Moreover, this review tried to give an overview on the research has so far done in Asian countries particularly in the field of natural antioxidants during the period 2000-2008. Therefore, the aim of this review is to present a comprehensive index for the antioxidant property of herbs and plants that have been traditionally used as well as experimentally proved beneficial in these countries. Approach: We had compared Asian and other nation's diet in the popularity of the traditional medicine and the type of daily meal in respect to their antioxidants contents and valuable health promoting effects. Many literatures had been collected using online search engines and the main library of University Putra Malaysia. The gathered data had been arranged alphabetically in a table depending on the plant's scientific name, common name, the used part(s) and the family name. Results: The research had shown that there were seventy-six plants with proved antioxidant activity which had been well studied by Asian scientists in the chosen period of time. Conclusion: The antioxidant components of the natural products constitute the major source of human health promotion and maintenance. The nature is still the perfect source for health promotion and for the supplementation of safe drugs. Great attention is mandatory from all the scientists around the world to explore many underestimated plants with highly effective antioxidant activity

Tryptophan Stimulates Immune Response in Broiler Chickens Challenged with Infectious Bursal Disease Vaccine

Infectious bursal disease is still a challenging issue by posing a serious threat to the commercial poultry industry especially due to the emergence of highly Infectious Bursal Disease Virus (IBDV). In the present study, we evaluated the immunomodulatory effects of Tryptophan (Tip) on innate, humoral and cellular immune responses in chickens challenged by oral administration of intermediate plus strain of IBD virus at 28 days of age. A corn-soybean meal based diet containing different levels of Tip (0, 0.10 and 0.20) for the starter, (0, 0.07 and 0.15) for the grower and (0, 0.05 and 0.13) for the finisher has been utilized. In a completely randomized design with three treatments of five replicates each and 10 chickens per replicate, 150 Cobb 500 male broiler chickens from 0-49 days of age were subjected to Tip diet. To measure the innate, cellular and humoral immunity indicators (interferon-alpha, interferon-gamma, immunoglobulin G, respectively) at 27, 35, 42 and 49 days of age, serum samples from each replicate of treatments were collected and subjected to ELISA. The result showed that Trp supplementation in the chickens basal diets significantly increased the serum levels of interferon-alpha at 35, 42 and 49 days of age (p<0.05), interferon-gamma at 27, 35 and 49 days of age (p<0.05) and immunoglobulin G at 27, 35, 42 and 49 days of age (p<0.05). These results strongly suggest that tryptophan plays a vital role in modulation of protective immune response against IBDV

Nutrition and immunity : the effects of the combination of arginine and tryptophan on growth performance, serum parameters and immune response in broiler chickens challenged with infectious bursal disease vaccine

To explore the effects of the combination of tryptophan (Trp) and arginine (Arg) on growth performance, serum parameters and immune response of broiler chickens challenged with intermediate plus strain of infectious bursal disease virus vaccine, an in vivo experiment was conducted. A corn-soybean meal-based diet containing different levels of Arg and Trp was used. Cobb500 male broiler chickens from 0 to 49 days of age were subjected to a diet supplemented with the combination of Trp and Arg. Growth performance parameters and serum parameters were measured at 27 and 49 days of age. To evaluate the immunomodulatory effects of the combination of Trp and Arg on the challenged chickens, we measured the serum levels of interferon-α, interferon-γ and immunoglobulin G at 27, 35, 42, and 49 days of age. The results showed that the three evaluated immune system parameters including interferon-α, interferon-γ and immunoglobulin G were significantly enhanced after treatment. This enhancement resulted in the recovery of infectious bursal disease virus-infected chickens compared with controls as confirmed by histopathological examinations. Moreover, serum parameters such as albumin and total protein increased, whereas the treatment decreased (P<0.05) the feed:gain ratio, aspartate amino-transferase, alkaline phosphatase, lactic dehydrogenase, triglyceride and cholesterol. These findings suggest that the combination of Arg and Trp has a regulatory effect on growth performance. Moreover, it modulates the systemic immune response against infectious bursal disease

Growth performance and blood parameters as influenced by different levels of dietary Arginine in broiler chickens.

An experiment was conducted to determine effects of dietary Arginine (ARG) on growth performance and blood serum parameters in broiler chickens. A corn-soybean meal based diet containing different levels of ARG (0, 0.67, 1.37, 2.07 and 2.77) for the starter (0, 0.53, 1.1, 1.68 and 2.25) for the grower and (0, 0.52, 1.04, 1.56 and 2.08) for the finisher was used. In a completely randomized design with five treatments of five replicates each and 10 chickens per replicate, 250 Cobb 500 male broiler chickens from 0-49 days of age were used. Growth performance (body weight gain, feed intake and feed: gain ratio) and blood serum (albumin, total protein, glucose, cholesterol, triglyceride, urea, uric acid, aspartate amino-transferase, alanine amino-transferase, alkalin phosphatase, lactic dehydrogenase and creatine kinase) parameters were measured at 27 and 49 days of age. Increase of dietary ARG increased (p0.05) aspartate amino-transferase and cholesterol. It was concluded that dietary ARG might have positive effects on health status of the broiler chickens

Identification and characterisation of a novel anti-viral peptide against avian influenza virus H9N2

Background: Avian influenza viruses (AIV) cause high morbidity and mortality among the poultry worldwide. Their highly mutative nature often results in the emergence of drug resistant strains, which have the potential of causing a pandemic. The virus has two immunologically important glycoproteins, hemagglutinin (HA), neuraminidase (NA), and one ion channel protein M2 which are the most important targets for drug discovery, on its surface. In order to identify a peptide-based virus inhibitor against any of these surface proteins, a disulfide constrained heptapeptide phage display library was biopanned against purified AIV sub-type H9N2 virus particles. Results: After four rounds of panning, four different fusion phages were identified. Among the four, the phage displaying the peptide NDFRSKT possessed good anti-viral properties in vitro and in ovo. Further, this peptide inhibited the hemagglutination activity of the viruses but showed very little and no effect on neuraminidase and hemolytic activities respectively. The phage-antibody competition assay proved that the peptide competed with anti-influenza H9N2 antibodies for the binding sites. Based on yeast two-hybrid assay, we observed that the peptide inhibited the viral replication by interacting with the HA protein and this observation was further confirmed by co-immunoprecipitation. Conclusion: Our findings show that we have successfully identified a novel antiviral peptide against avian influenza virus H9N2 which act by binding with the hemagglutination protein of the virus. The broad spectrum activity of the peptide molecule against various subtypes of the avian and human influenza viruses and its comparative efficiency against currently available anti-influenza drugs are yet to be explored

Inhibition of Growth of Highly Resistant Bacterial and Fungal Pathogens by a Natural Product

The continuous escalation of resistant bacteria against a wide range of antibiotics necessitates discovering novel unconventional sources of antibiotics. B. oleracea L (red cabbage) is health-promoting food with proven anticancer and anti-inflammatory activities. However, it has not been researched adequately for its antimicrobial activity on potential resistant pathogens. The methanol crude extract of B. oleracea L. was investigated for a possible anti-microbial activity. The screening method was conducted using disc diffusion assay against 22 pathogenic bacteria and fungi. It was followed by evaluation of the minimum inhibitory concentration (MIC). Moreover, the antibacterial and the antifungal activities were confirmed using the minimum bactericidal concentration (MBC) and the minimum fungicidal concentration (MFC), respectively. Remarkable, antibacterial activity was evident particularly against highly infectious microorganisms such as Methicillin-resistant Staphylococcus aureus, Escherichia coli O157:H7, Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus, and Salmonella enterica serovar Typhimurium as well as against human fungal pathogens, Trichophyton rubrum and Aspergillus terreus. Red cabbage is a rich source of phenolic compounds, anthocyanins being the most abundant class, which might explain its potent antimicrobial action. This extract is potentially novel for future antimicrobials, inexpensive, and readily available at a large scale for pharmaceutical companies for further investigation and processing

Dietary Tryptophan Effects on Growth Performance and Blood Parameters in Broiler Chicks

In order to study the effect of Tryptophan (Tip) on growth performance and blood parameters of broiler chickens, an in vivo experiment was conducted. A corn-soybean meal based diet containing different levels of Trp (0, 0.10 and 0.20) for the starter, (0, 0.07 and 0.15) for the grower and (0, 0.05 and 0.13) for the finisher was used. In a completely randomized design with three treatments of five replicates each and 10 chickens per replicate, 150 Cobb500 male broiler chickens from 0-49 days of age were subjected to Trp diet. Growth performance (body weight gain, feed intake and feed, gain ratio) and blood serum (albumin, total protein, glucose, cholesterol, triglyceride, urea, uric acid, aspartate amino-transferase, alanine amino-transferase, alkalin phosphatase, lactic dehydrogenase and creatine. e kinase) parameters were measured at 27 and 49 days of age. As the result showed the increase of dietary Trp elevated (p 0.05) body weight gain, feed intake, albumin, total protein, glucose, urea and uric acid and decreased (p>0.05) feed gain ratio, aspartate amino-transferase, lactic dehydrogenase, triglyceride and cholesterol. Therefore, we conclude that dietary Tip might have positive effects on health status of the broiler chickens

Bioactive compounds and antioxidant activity of different extracts from Vitex negundo leaf

The study was conducted to assess the antioxidant activity of methanol and hexane extract and essential oil from Vitex negundo leaf using different in vitro antioxidant assays. Antioxidant property was tested using 2,2-diphenyl-1-picrylhydrozyl (DPPH) free radical scavenging capacity, ferric ion reducing antioxidant power (FRAP) and β-carotene-linoleic acid assays. Total phenolic contents (TPC) were measured using Folin-Ciocalteu method. Flavonoids, tocopherols, β-carotene and lycopene were analyzed using high performance liquid chromatography (HPLC). Results of the present study showed that methanol extract of V. negundo leaf exhibited significantly (p < 0.05) higher antioxidant activity in terms of measurements of DPPH free radical (IC50), FRAP and β-carotene-linoleic assays than those of hexane extract and essential oil. Methanol extract of V. negundo leaf also contained high amounts of bioactive compounds including total phenolic compounds (363 mg GAE/g), epicatechin (16.98 mg/g), quercetin (13.45 mg/g), catechin (8.95 mg/g) and myricetin (3.32 mg/g) while the concentrations of tocopherol, β-carotene and lycopene were found to be lower