Open Access Original Research Article

Biomarker Analysis with Grating Coupled Surface Plasmon Coupled Fluorescence

Alvaro Mendoza, James A. Dias, Theodore Zeltner, David A. Lawrence

Journal of Advances in Biology & Biotechnology, Page 1-22
DOI: 10.9734/JABB/2014/11452

One of the greatest hurdles in protein microarray technology has been the limited sensitivity. However, the strategy of amplifying the SPR signal with fluorescence improves sensitivity. Herein, we report a simple and efficient method, Grating Coupled Surface Plasmon Coupled Fluorescence (GCSPCF) with a peptide microarray to screen sera of human subjects and a mouse strain with autistic-like behavior for quantification of the epitope specificities of their circulating antibodies. The GCSPCF peptide microarray is a novel assay that facilitates high-throughput screening of antibody epitope specificities or binding of interacting analytes with a small quantity (<50 µl) of sample. This study utilized over 600 peptides with the eventual goal of being able to identify antibodies that could be used as biomarkers of particular disorders. The GCSPCF technology was able to detect antibodies to the tripeptide glutathione; it was also capable of sequential screening antibodies of different immunoglobulin isotypes (IgG1, IgG4 and IgE) to the same peptide. The GCSPCF broad-based screening approach was able to begin the identification of epitope-specific antibody binding patterns in an effort to develop a predictive signature that could be further developed for clinical trials. In addition to evaluation of antibody specificities with hundreds of spotted peptides at multiple regions of interest (ROI), the addition of spotted antibodies could provide an even more complete biomarker profile with capture of analytes, such as stress proteins and cytokines. Differences between a mouse strain with normal behaviors (C57BL/6J) and a strain with autistic-like behaviors (BTBR T+tf/J) as well as healthy and autistic participants are used for comparative analyses.

Open Access Original Research Article

Analgesic (Antinociceptive) Property of Moringa oleifera Ethanol Leaf Extract in Albino Rats

Ijioma N. Solomon, Madubuike G. Kelechi, Nwosu O. Chinenyenwa

Journal of Advances in Biology & Biotechnology, Page 23-29
DOI: 10.9734/JABB/2014/11679

Aim: This study investigated the analgesic property of ethanol leaf extract of Moringa oleifera (ELMO) in rats.

Study Design: Different rat models (acetic acid, tail flick and tail immersion) were employed. The study was carried out in the Physiology Laboratory of the Department of Veterinary Physiology, Pharmacology, Biochemistry and Animal Health, College of Veterinary Medicine, Michael Okpara University of Agriculture, Umudike, Nigeria, between July and December, 2013.

Methodology: 42 albino Rats were used for each experimental model. Varying doses of ELMO: 200, 300, 400 and 500 mg/kg were administered orally to rats in different groups. The negative control group received 0.2ml normal saline while Aspirin (10mg/kg) and Dicloferanc Potassium (2mg/kg) were used as reference drugs.

Results: ELMO exerted a dose dependent reduction in number of writhes per time in the acetic acid model as the varying doses used inhibited the number of writhes observed in the negative control group by 33.33, 57.84, 67.16 and 66.67% respectively. ELMO also increased reaction times in the tail flick and tail immersion models with 400mg/kg producing inhibition ratios of 10.05 and 11.51 respectively. Results in the treated groups were significantly (P<0.05) different from that of the negative control group and compared favorably with those of Aspirin and Diclofenac Potassium.

Conclusion: The experiments hence indicate that ELMO could contain principles with strong analgesic properties and seems promising in the search for new analgesic agents with minimal side effects and high potency for the relief of all forms of pain.

Open Access Original Research Article

Comparative Physical Characterization, Physio-Chemical and Fatty Acid Composition of Some Edible Vegetable Oils

N. M. Essien, O. E. Ofem, S. C. Bassey

Journal of Advances in Biology & Biotechnology, Page 30-39
DOI: 10.9734/JABB/2014/12489

Edible vegetable oils which include red palm oil, coconut oil, groundnut oil, sesame oil (Beniseed oil), soybean oil, palm kernel oil, etc. are derived from seeds or fruits of different plants. These oils are consumed not only for their supply of lipids in the diets but for their distinct aromas, colours, palatability and availability. Vegetable oils are rich in essential nutrients such as vitamins and anti-oxidant compounds. The type of diet and in particular the nature of dietary fats has been found to raise or lower the blood cholesterol in man. This study was therefore necessary as it will assess and ascertain the physiochemical and fatty acid characteristics of the various vegetable oils available in the Nigerian markets. The various physiochemical and fatty acid parameters were estimated using standard procedure. Results indicate that red palm oil had high levels of palmitic and oleic acids as well as peroxide and iodine values, with high fire and boiling points. Coconut oil had high peroxide, saponification and acid values, high concentrations of capric, lauric, palmitic, myristic, stearic and linoleic acids with high smoke point. Palm kernel oil had high iodine, saponification and acid values, high contents of capric, lauric and myristic acids with high flash, fire and boiling points. Oleic and stearic acids were in turn very high in soybean, sesame. In conclusion, results indicate that red palm oil, soybean and sesame oil would be safer for consumption since there are less atherogenic compared with the other vegetable oils.

Open Access Original Research Article

Screening, Optimization and Extraction of Polyhydroxyalkanoates (PHAs) from Bacillus thuringienesis

S. E. Desouky, H. H. El- Shiekh, M. A. Elabd, A. M. Shehab

Journal of Advances in Biology & Biotechnology, Page 40-54
DOI: 10.9734/JABB/2014/12286

Aims: This study illustrated obtaining bacterial isolates capable to produce polyhydroxyalkanoates (PHA) under stressed conditions and also using cheap wastes for production process.

Study Design: Two-step throughput screening of 50 bacterial isolates by lipophylic stain (Sudan black B) and flouroscence stain (Nile red) will be carried on, then the best condition for PHA production followed by identification of most potent isolate by molecular charterization will be investigated.

Place and Duration of Study: The study were performed in physiological lab in botany & microbiology department in faculty of science, Al-azhar university and NRC from October 2012 until April 2014.

Methodology: Bacterial isolates will be investigated using two different indicator stains, Sudan Black and Nile Red, for PHA productivity then the highly producing isolate will be identified. Factors controlling PHA production and stressed condition will be studied alternatively depends on increasing of PHA production which will be assayed quantitatively by converting PHA to crotonic acid. Biopolymer will be extracted using successive solvents.

Result: Five soil samples were collected from different localities contaminated with industrial wastes and 50 isolates of bacteria were purified then screened for PHA production by using Sudan black B as a primary screening and confirmed by fluorescent Nile red staining. Eleven promising bacterial isolates were selected based on their PHA yields. Out of eleven natural promising isolates, BS11 was found to be the most efficient PHA producer and identified as Bacillus thuringiensis (KJ206079). The effect of different conditions on PHA produced by B. thuringiensis including carbon sources, nitrogen sources, incubation temperatures, pH and incubation periods were studied. The highest amount of PHA was obtained from this isolate using medium containing 30 g/l molasses, 0.8 g/L ammonium sulphate at pH 7.5 and incubation temp at 35ºC and after 72 hours. In conclusion, after the optimization of PHA synthesizing conditions, B. thuringiensis accumulate up to 69 % of its dry weight and can be employed for industrial production.

Conclusion: The experiment indicate the efficiency of two-throughput screening systems targeting PHA producing isolates. Also optmiziation of PHA producing factors for the most potent producing strain, Bacillus thuringiensis (KJ206079), was increased from 2.5 g/L to 4.1 g/L and represent about 69% cell dry weight.