Development Of An Indirect Elisa Using The Recombinant Nucleocapsid Protein As Antigen For Detection Of Antibodies To Newcastle Disease Virus

Newcastle disease (ND) is a virulent form of poultry disease that has the potential to cause 100% mobidity and mortality. The continuing threat of ND to the poultry industry requires routine testing of vaccinated chickens to determine that they have been adequately immunised by vaccination. Enzyme-linked immunosorbent assays (ELISA) which use commercially available reagents are presented as alternatives to the procedure for detection of Newcastle disease virus (NDV) antibodies. The nucleocapsid protein has been shown to immunogenic in nature, and play key roles in diagnostic ELISA for detection of antibody. A purified recombinant nucleocapsid protein (NP) of NDV highly expressed in Escherichia coli (E. coli) was used as the coating antigen in an indirect ELISA to detect the presence of NDV antibodies in a cohort of chicken sera. The test was standardised using sera from vaccination study and obtained from Charles River SPAFAS Laboratories, USA. The NPELISA was standardised at an antigen concentration of 1.5 μg/ml, test serum dilution of 1:200 and conjugate dilution of 1:2000. The cut-off value of the NP-ELISA of 0.238 was determined by a receiver operating characteristic (ROC) analysis. An assay of 315 chicken serum samples against NDV antibodies showed that the NP-ELISA had 92.06% sensitivity and 93.2% specificity compared to NDV hemagglutination–inhibition (HI) test. This was in good agreement between the two serological methods (κ =0.848). In the serum neutralization test (SNT), the result shows that there was no correlation observed between HI and SNT. It was observed that the field serum samples found positive or with high titer in the HI test were negative or have low titer for SNT. This suggests that different epitopes on the antigen have been recognized by each of the tests. The result shows that the recombinant NP antigen is recognized by antibodies specific to the other poultry pathogens and the cross reactivity is high for Paramyxovirus-2 (PMV-2) and Paramyxovirus-3 (PMV-3) but low reaction by other poultry pathogen [avian influenza (AI), avian encephalomyelitis (AE), infectious laryngotracheitis (ILT), infectious bronchitis (IB), and infectious bursal disease (IBD)]. The titre of ND antibodies can be determined using NPELISA formula, Log10Titre = 1.0* Log(S/P) + 3.45. This formula was derived after getting the linearity graph between NP-ELISA to commercial ND test kit. This finding indicates the potential application of the E. coli produced NP protein as antigen in indirect ELISA for the detection of NDV antibody. The NP-ELISA was successfully developed and very useful which is more or as efficient as commercial kits, thus the cost of NP-ELISA is cheaper than commercial kits

PSA campaign outline / Norsharina Samsuri

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An integrated formulation of zernike invariant for mining insect images

This paper presents mathematical integration of Zernike Moments and United Moment Invariant for extracting printed insect images.These features are further mining for granular information by investigating the variance of Interclass and intra-class. The results reveal that the proposed integrated formulation yield better analysis compared to convectional Zernike moments and United Moment Invariant

An Integrated Formulation of Zernike Invariant for Mining Insect Images

This paper presents mathematical integration of Zernike Moments and United Moment Invariant for extracting printed insect images.  These features are further mining for granular information by investigating the variance  of Interclass and intra-class. The results reveal that the proposed integrated formulation yield better analysis compared to conventional Zernike moments and United Moment Invarian

Thymoquinone rich fraction from Nigella sativa and thymoquinone are cytotoxic towards colon and leukemic carcinoma cell lines.

Nigella sativa has been used for centuries in Asia, Middle East and Africa to promote health and fight diseases. In this study, the anti-cancer effects of thymoquinone rich fraction (TQRF) extracted from N. sativa seeds using supercritical fluid extraction (SFE) system and commercially available thymoquinone (TQ) on colon cancer (HT29), lymphoblastic leukemia (CEMSS) and promyelocytic leukemia (HL60) cells lines were investigated. The concentration that gave 50% inhibition of cell viability (IC50) of HT29, CEMSS and HL60 cells treated with TQRF were 400, 350 and 250 µg/ml, respectively. Meanwhile, the IC50 of TQ was 8, 5 and 3 µg/ml, respectively. Cell cycle analysis shows the increment of apoptosis in a time-dependent manner. However, both TQRF and TQ were not able to arrest the cell cycle phases of the cells. Apoptosis was the main mode of HT29, CEMSS and HL60 cells death induced by both TQRF and TQ. Our findings support the potential use of TQRF and TQ for the treatment of colon cancer and leukemia

Antioxidant activity and phenolic content of phenolic rich fractions obtained from black cumin (Nigella sativa) seedcake.

The antioxidant activities of crude methanolic extract (CME) and its fractions using ethyl acetate (EAF), hexane (HF) and water (WF) of black cumin seedcake were investigated. DPPH radical scavenging activity, β-carotene-linoleate bleaching, and inhibition of corn oil oxidation were used to evaluate the antioxidant capacity. The total phenolics were found to be 78.8, 27.8, 32.1 and 12.1 mg gallic acid equivalents (GAE)/g in EAF, CME, WF and HF, respectively. The CME and EAF exhibited the highest DPPH followed by WF and HF. The extract/fractions showed high effect on reducing the oxidation of β-carotene. The effect of extract/fractions on the oxidative stability of corn oil at 70 °C was tested in the dark and compared with butylated hydroxyanisole (BHA). The oil peroxide and anisidine values were generally lower with addition of PRFs in comparison to a control. The predominant phenolic compounds identified by HPLC-DAD in CME and WF of black cumin seedcake were hydroxybenzoic, syringic and p-cumaric acids

Antioxidant activities of phenolic rich fractions (PRFs) obtained from black mahlab (Monechma ciliatum) and white mahlab (Prunus mahaleb) seedcakes

The antioxidant activities of phenolic rich fractions (PRFs) from crude methanolic extract (CME), and its fractions using ethyl acetate (EAF), hexane (HF) and water (WF) of black mahlab (Monechma ciliatum) and white mahlab (Prunus mahaleb) seedcakes were investigated. The total phenolic compounds were found to be higher in white mahlab than black mahlab seedcakes. The antioxidant activity determined by the DPPH method revealed that black mahlab PRFs had the highest antioxidant activity, compared to white mahlab fractions. The presence of antioxidants in the two mahlab PRFs reduced the oxidation of β-carotene by hydroperoxides from these extracts/fractions. The effect of the two mahlab PRFs on the oxidative stability of corn oil at 70 °C was tested in the dark and compared with butylated hydroxyanisole (BHA). The CME performed better antioxidant activity in inhibiting the formation of both primary and secondary oxidation products. The qualitative and quantitative characterisation of phenolic compounds was carried out by HPLC/DAD

Ethyl acetate extract of germinated brown rice attenuates hydrogen peroxide-induced oxidative stress in human SH-SY5Y neuroblastoma cells: role of anti-apoptotic, pro-survival and antioxidant genes

Background There are reports of improved metabolic outcomes due to consumption of germinated brown rice (GBR). Many of the functional effects of GBR can be linked to its high amounts of antioxidants. Interestingly, dietary components with high antioxidants have shown promise in the prevention of neurodegenerative diseases like Alzheimer’s disease (AD). This effect of dietary components is mostly based on their ability to prevent apoptosis, which is believed to link oxidative damage to pathological changes in AD. In view of the rich antioxidant content of GBR, we studied its potential to modulate processes leading up to AD. Methods The total phenolic content and antioxidant capacity of the ethyl acetate extract of GBR were compared to that of brown rice (BR), and the cytotoxicity of both extracts were determined on human SH-SY5Y neuronal cells using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) Assay. Based on its higher antioxidant potentials, the effect of the GBR extract on morphological changes due to hydrogen peroxide (H2O2)-induced oxidative damage in human SH-SY5Y neuronal cells was examined using inverted light microscope and fluorescence microscope by means of acridine orange-propidium iodide (AO/PI) staining. Also, evaluation of the transcriptional regulation of antioxidant and apoptotic genes was carried out using Multiplex Gene Expression System. Results The ethyl acetate extract of GBR had higher total phenolic content and antioxidant capacity compared to BR. The cytotoxicity results showed that GBR extract did not cause any damage to the human SH-SY5Y neuronal cells at concentrations of up to 20 ppm, and the morphological analyses showed that the GBR extract (up to 10 ppm) prevented H2O2-induced apoptotic changes in the cells. Furthermore, multiplex gene expression analyses showed that the protection of the cells by the GBR extract was linked to its ability to induce transcriptional changes in antioxidant (SOD 1, SOD 2 and catalase) and apoptotic (AKT, NF-Kβ, ERK1/2, JNK, p53 and p38 MAPK) genes that tended towards survival. Conclusions Taken together, the results of our study showed that the ethyl acetate extract of GBR, with high antioxidant potentials, could prevent H2O2-induced oxidative damage in SH-SY5Y cells. The potential of GBR and its neuroprotective mechanism in ameliorating oxidative stress-related cytotoxicity is therefore worth exploring further

Toxicology study of vanillin on rats via oral and intra-peritoneal administration.

Vanillin is useful as anti-sickle cell anemia, anti-mutagen and anti-bacteria agent. However, vanillin must be administered at high concentration and cannot be oxidized by the upper gastrointestinal track of patients to be medically effective. In this study, we assessed the toxic effect of vanillin when administered in an un-oxidized form at high concentrations (150 and 300 mg/kg) via oral and intra-peritoneal injection. It was found that 300 mg/kg vanillin injection caused the rats to be unconscious without exerting any toxic effect on blood cells, kidney and liver. Besides, it showed blood protective property. Further analysis with GenomeLab GeXP genetic system on brain tissues showed that the expression of most xenobiotic metabolism, cell progression, tumor suppressor, DNA damage and inflammation genes were maintained at normal level. However, the expression of a few xenobiotic metabolism, cell cycle arrest and apoptosis genes were up-regulated by 5% ethanol injection. Nevertheless, when 5% ethanol was injected with the presence of vanillin, the expression was back to normal level. It is postulated that vanillin might have neuro-protective property. In conclusion, vanillin is not toxic at high concentration in both oral and intra-peritoneal injection and could provide blood and brain protective properties

Polyphenol-rich ethyl acetate fraction of Molineria latifolia rhizome restores oxidant-antioxidant balance by possible engagement of KEAP1-NRF2 and PKC/NF-κB signalling pathways

The present study examined the potential of ethyl acetate fraction (EAF) isolated from Molineria latifolia rhizome to modulate the oxidant-antioxidant balance in high fat diet and nicotinamide/streptozotocin-induced diabetic rats. Administration with EAF ameliorated systemic antioxidant status- and oxidative stress-related parameters without affecting the renal and liver functions in the diabetic rats. Further analyses on adipose, muscle and liver tissues demonstrated differing ability to scavenge free radicals and protection against lipid peroxidation. Transcriptional changes proposed concerted modulation of both KEAP1-NRF2 and PKC/NF-κB signallings in tissue-specific manner. Qualitative profiling of compounds present in EAF was analysed by non-targeted HPLC-QTOF mass spectrometry. A total of 23 unique mass signals were detected in EAF. Putative identification revealed a mixture of naturally occurring polyphenols ranging from cinnamic acid-, benzoic acid- and flavonoid-derived groups. Overall, the study demonstrated potential application of EAF to reinstate diabetes-induced oxidant-antioxidant imbalance by potentially modulating the NRF2–NF-κB signalling axis