Open Access Method Article

A Comprehensive Analysis of the Microarray Gene Expression Protocol and Results Using the Mouse T-cell Gene Expression Profile

Onoriode Oyiborhoro, Oriakhi Kelly, Esosa S. Uhunmwangho, Kingsley A. Iteire, Enoh F. Akpojotor

Asian Journal of Biochemistry, Genetics and Molecular Biology, Page 1-15
DOI: 10.9734/ajbgmb/2019/v2i430069

The microarray technology is a very powerful technology that combines molecular biology and computer technology to analyze the gene expression levels for most or all of the genes in a whole genome simultaneously, at very high resolutions. This technology has wide applications, including gene interaction studies for discovery of genes responsible for different diseases; classification of cancers and other diseases; prediction of clinical outcomes or prognosis for different diseases; response to therapy and development of new therapeutic agents, including gene therapy. It is therefore a very potent, unbiased and sensitive technology for the discovery of novel genes involved in the pathogenesis or control of diseases including cancers and autoimmune diseases. In the present study, we seek to give a clear and detailed account of the microarray gene expression protocol using the mouse T-cell gene expression profile, including challenges involved and how to overcome them, as well as detailed analysis of results obtained.

Open Access Original Research Article

Biochemical Changes Associated with Consumption (by Rats) of “Garri” Processed by Traditional and Instant Mechanical Methods

A. I. Airaodion, A. C. Ene, E. O. Ogbuagu, V. N. Okoroukwu, J. A. Ekenjoku, U. Ogbuagu

Asian Journal of Biochemistry, Genetics and Molecular Biology, Page 1-11
DOI: 10.9734/ajbgmb/2019/v2i430064

Aim: This study aimed to investigate the biochemical changes in rats, associated with the consumption of “garri” (roasted (fried) fermented cassava flour) processed by traditional and instant mechanical methods.

Methods: Cassava samples obtained from the International Institute of Tropical Agriculture (IITA), Ibadan were processed using traditional and instant mechanical methods. Fifteen adult male Wistar rats were purchased from the Animal Holding Unit of the Department of Physiology, University of Ibadan, Nigeria, with body weights between 100 and 120 g. They were acclimatized for 7 days during which they were fed ad libitum with standard feed and drinking water. They were randomly divided into 3 groups of 5 rats each. Rats in group A were fed with pure standard rat chow. Rats in group B were fed with garri processed by instant mechanical method (fermented ≤ 24 hours before roasting) while those in group C were fed with garri processed by the traditional method (fermented ≥ 72 hours before roasting). After 28 days of feeding, the animals were fasted overnight and anaesthetized using diethyl ether. Blood samples were collected by cardiac puncture. Hepatic and renal indices were determined using standard methods.

Results: Mild perturbations were observed in the liver and renal biochemical parameters when animals fed with garri processed by the traditional method were compared with that fed garri processed by instant mechanical method and control diet. These perturbations were more severe in animals fed with garri processed by instant mechanical method.

Conclusion: In this study, both garri samples did not cause hepatic nor renal damage but mild perturbation of biochemical parameters were observed. These perturbations were more severe in animals fed with garri processed by instant mechanical method. This could be attributed to the high cyanide content in it.

Open Access Original Research Article

Extraction, Purification and Kinetic Study of Lactate Dehydrogenase of Male Chicken from Ebocha-oil Exploration Area, Nigeria

Chimdi E. Esonu, G. O. C. Onyeze, Kizito M. E. Iheanacho, Linus N. Nwaogu, Simon-Peter Odirichukwu

Asian Journal of Biochemistry, Genetics and Molecular Biology, Page 1-12
DOI: 10.9734/ajbgmb/2019/v2i430065

Aim: This study focused on the extraction, purification and kinetic studies of lactate dehydrogenase of male chickens from Ebocha oil exploration area, Imo state, Nigeria.

Methods: Twenty-one apparently healthy mature (6-9 months) male chickens (Gallus domesticus) from Ebocha oil exploration area, Imo State, Nigeria were screened for lactate dehydrogenase activity, thus accessing the level of chronic cell exposure to gas flaring. Their thigh muscle tissues were severed and investigated for lactate dehydrogenase activity using the standard method and sodium pyruvate as the substrate. Lactate dehydrogenase was isolated and purified by ammonium sulphate precipitation, desalted by dialysis and then gel filtration.

Results: The enzyme activity increased with advancement in the purification steps and was maximum using dialysis. The values for the lactate dehydrogenase activities were 103.43±3.27 U/L, 279.50±5.38 U/L, 318.16±13.08 U/L, 100.47±2.59 U/L, with a purification fold of 1, 3.7, 6.24 and 2.55 for the purification steps respectively. Also, the values of the protein concentrations were 0.071 mg/ml, 0.050 mg/ml, 0.035 mg/ml and 0.027 mg/ml (values for the crude enzyme, ammonium sulphate precipitation, dialysis and gel filtration respectively). The enzyme showed optimal activity at pH range of 5.5-6.5 and temperature of 30ºC-40ºC. Using sodium pyruvate as the substrate, with a fixed enzyme volume, an increase in the concentration of substrate resulted in increase in enzyme activity until a saturation point 0.3mM was reached. The apparent Km and Vmax values obtained were 0.01 mM and 0.12 U/mg/min. The Lineweaver-burk plot of the partially purified enzyme gave real Km and Vmax values of 0.20 mM and 0.16 U/mg/min respectively.

Conclusion: Partial purification procedures and biochemical properties of lactate dehydrogenase, from the muscle tissues of male chickens of Ebocha origin, gives room for more investigation on the metabolic shift caused by chronic exposure of the environment, humans and livestock to gas flaring and petroleum exploration.

Open Access Original Research Article

Effects of Powdered Stem Bark of Terminalia avicennioides Made as Dietary Feed Fed to Mice Infected with Plasmodium berghei, on Liver Function

Afolabi Owoloye, Olusegun Matthew Akanbi, Oluwafemi Shittu Bakare

Asian Journal of Biochemistry, Genetics and Molecular Biology, Page 1-11
DOI: 10.9734/ajbgmb/2019/v2i430066

Aim: This study aimed to investigate the antiplasmodial activity and effect of stem bark of Terminalia avicennioides made as dietary feed fed to mice infected with Plasmodium berghei, on some serum biochemistry.

Methodology: Twenty (20) mice were divided into four groups. Group 1 was not infected with Plasmodium berghei (normal control), Group 2 was infected with P. berghei but not treated (negative control). Group 3 was infected and treated with 5.0 mg/kg of Arthemeter-Lumefantrine (positive control). Groups 4 was infected and fed with treated feed (T. avicennioides). Treatments were carried out for five days. Blood was taken daily from the tail of the mice before treatment for the assessment of parasitaemia. The animals were sacrificed on the fifth day and the whole blood was collected into EDTA bottle. Serum obtained was used to assay for biochemical parameters.

Results: Parasitaemia count was significantly lower (p<0.05) in all the treated groups when compared with the negative control group. The high-density lipoprotein was significantly higher (P<0.05) in the normal control (123.14±3.19) when compared with the positive control (99.18±2.76), negative control (85.29±0.85) and the group treated with T avicennioides (86.14±3.21). The serum alanine aminotransferase, alkaline phosphatase and aspartate aminotransferase level in the group treated with T. avicennioides (167.90±4.13, 15.87±1.32 and 17.50±1.95) respectively were significantly reduced (p<0.05) when compared with the negative control (197.25±5.44, 20.01±1.32 and 26.71±0.45) respectively. The mean bilirubin and albumin level in the negative control showed no significant difference when compared with the group fed with T. avicennioides.

Conclusion: The study concluded that T. avicennioides have antiplasmodial activity with a mild adverse effect on liver function.

Open Access Original Research Article

Biochemical Characterization on Photosynthetic Activities during Dark Incubated Senescence of Wheat Primary Leaves by Gibberellic Acid

G. Fareeda, S. D. S. Murthy

Asian Journal of Biochemistry, Genetics and Molecular Biology, Page 1-5
DOI: 10.9734/ajbgmb/2019/v2i430067

In annual crop plants like maize, rice and wheat etc. Senescence limits crop yields of annual crops like maize rice and wheat. Delayed leaf senescence is a desirable agronomic trait to improve crop yield. In this study 10 µM GA reduced the loss of wheat primary leaves under incubated dark conditions. GA reduced the loss of pigments, proteins, electron transport activities, spectral properties. The restoration of WCE activity by GA was closely associated with the restoration of PS II activity compared to that of PS I. GA treated leaf thylakoid membranes showed an increase in absorption at 680 nm moderate increase at 480 nm and 440 nm at 72 h during dark incubation. GA protected the degradation of water oxidation complex polypeptides (33, 23, 17 KDa) of PS II and slightly protected the PS I polypeptides.

Open Access Original Research Article

The Efficiency and Effectiveness of Open Pollination in Musa Breeding

Victoria Wilson, Abdou Tenkouano

Asian Journal of Biochemistry, Genetics and Molecular Biology, Page 1-15
DOI: 10.9734/ajbgmb/2019/v2i430068

Aims: This field experiment was conducted to determine if hand and open pollination methods affected performances of Musa progenies from 4x - 2x crosses and to identify promising progenies for recurrent selection.

Study Design: The experimental design was a randomized complete block design with two replications of 6 plants per genotype.

Place and Duration of Study: International Institute of Tropical Agriculture (IITA) High Rainfall Station, Onne (4º51’N, 7º03’E, 10 m above sea level), in Rivers State, South-south Nigeria for 24 months.

Methodology: Two-month old seedlings of hand pollinated (6 diploid, 6 tetraploid) and open pollinated (6 diploid, 6 tetraploid) progenies, along with parental clones (2x) and (4x) of each genotype were planted at 3 m x 2 m spacing. Data on phenology, vegetative growth, yield and yield characters were collected at flowering and harvest over three crop cycles. Genotypes were partitioned into 5 clusters assayed by means of orthogonal contrasts to compare the performance of progenies from both pollination methods.

Results: Pollination methods produced no significant (P = .05) differences, unfavourable effects or reduction in performance of economically important yield and yield components of 4x and 2x progenies of similar genotype. Some significant (P = .05) linear correlations and relationships between phenological and vegetative traits; and yield and yield components changed with pollination methods and ploidy levels but did not affect final outcomes. Promising open pollinated diploids include the early maturing TMP2x 2829-62OP; and for high yield and yield components measured, TMB2x 8084-2OP and TMP2x 1448-1OP. Promising open pollinated tetraploids include TMP4x 7002-1OP and TMP4x 2796-5OP.

Conclusion: Open pollination did not result in unfavorable effects or reduction in performance of economically important yield and yield components in progenies of similar genotypes. Therefore, open pollination could be considered for Musa breeding. This will reduce cost, labour, time and stress involved in Musa improvement.

Open Access Original Research Article

Studies on the Phytochemical Compounds in the Ethanolic Leaf Extract (ELE), Ethanolic Bark Extract (EBE) and Ethanolic Root Extract (ERE) of Bridelia ferruginea Benth (Euphorbiaceae)

Abdullahi Attah Alfa, Orukotan Abimbola Ayodeji, Goji Anthony Donatus Teru, Kokori Bajeh Tijani

Asian Journal of Biochemistry, Genetics and Molecular Biology, Page 1-8
DOI: 10.9734/ajbgmb/2019/v2i430070

The phytochemical compounds of Bridelia ferruginea plant parts was carried out using qualitative method to determine the bioactive compounds present in the plant leave, stem bark and root extracts. The samples was weighed, of which 100 g each of the powder were extracted in solvents (ethanol) 1000 ml macerated and stand for 72 hours. The solvents contained in the maceration bottle was decanted and filtered using a filter paper, the filtration was aided using a suction pump. The filtrate was concentrated using a rotary evaporator and then transferred into thermostatic water cabinet (Temperature was set at 45oC), allowed to dry completely. The plant parts extracts were separately kept in a screw capped bottle for further research. The bioactive compound in the plant parts were detected. The result revealed that Carbohydrates, Saponins, Flavonoids, Tannins, Cardiac Glycosides, fats and oils were present. Alkaloid present in Dragendoff’s test in all plant parts extract but absent in Mayer’s test in only leaf extract. Terpenoids/Steroids present in Liebermann-Burchard’s test in all plant parts extract but absent in Salkowski’s test in only leaf extract. Anthraquinones were absent in all plant parts extracts using Bontrager’s test. Therefore, the presence of these phyto-pharmacological compounds is an indicative that the plant is medicinal and it can be used for the treatment of bacterial and other microbial infections. Further study can be done to separate the individual metabolites to test their antimicrobial activity against some pathogenic bacteria like bacterial meningitis, tuberculosis and syphilis to determine their potency.

Open Access Original Research Article

Paternal Contribution to Banana (Musa sapientum L.) & Plantain (Musa paradisiaca L.) Progenies & Progeny Ploidy Composition in a Polycross Mating System Using RAPD & Flow Cytometry

Victoria Wilson, Abdou Tenkouano, Michael Pillay

Asian Journal of Biochemistry, Genetics and Molecular Biology, Page 1-14
DOI: 10.9734/ajbgmb/2019/v2i430071

Aims: A 4x – 2x polycross mating design of 4 tetraploid female parents was established to determine paternal contributions of 3 diploid male parents to resulting progenies, their ploidy composition and genetic diversity of synthetic hybrids.

Study Design: The polycross mating design comprised 2 blocks having both maternal and paternal selections, with seed parents replicated at 12 plants per clone. Each crossing block had 31 plants of each of the three male parents.

Place and Duration of Study: International Institute of Tropical Agriculture (IITA) High Rainfall Station, Onne (4º51’N, 7º03’E, 10 m above sea level), Rivers State, South-South Nigeria for a period of 24 months.

Methodology: At maturity of maternal parents (TMPx 2796-5; TMPx 1658-4; TMPx 5511-2; and TMPx 7152-2), fruit bunches were harvested, ripened and the seeds extracted. Hard seeds obtained were germinated in vivo in seed trays and emerging seedlings transplanted to perforated nursery bags. At 12 weeks, DNA was extracted from candle leaf for RAPD analysis of 80 progenies and the 3 pollen parents. Ploidy status of progenies was determined using flow cytometry method.

Results: There was significant unequal paternal contribution to Musa polycross progenies with 3 maternal parents; TMPx 2796-5, TMPx 5511-2, and TMPx 1658-4. Two of the 3 paternal parents had progenies with all 4 maternal parents while TMB2x 5105-1 did not have any progeny with TMPx 2796-5. Progenies exhibited 4 ploidy levels with frequency differing with each female parent: TMPx 7152-2 produced 100% 3x progeny; TMPx 5511-2, 63% 3x and 37% 2x; TMPx 2796-5, 91% 3x and 9% 2x and TMPx 1658-4, 82% 3x, 9% 2x, 6% 4x and 3% 5x. The 5x progeny was recorded in the first ratoon crop. The second ratoon crop had only triploids.

Conclusion: The high frequency of 3x progenies from all maternal types in this study, suggests the effectiveness of the polycross mating design in Musa improvement.

Open Access Original Research Article

Effect of Silymarin on Cathepsin Activity and Oxidative Stress in TNBS-induced Colitis in Rats

Gokhan Bayramoglu, Hakan Senturk, Gungor Kanbak, Mediha Canbek, Aysegul Bayramoglu, Eda Dokumacioglu, Selin Engur

Asian Journal of Biochemistry, Genetics and Molecular Biology, Page 1-9
DOI: 10.9734/ajbgmb/2019/v2i430072

The purpose of the this study was to defined protective effects of silymarin on the experimental colitis induced by intra-colonic application of 2,4,6-trinitrobenzene sulfonic acid (TNBS).  The twenty eight Sprague-Dawley rats were randomly seperated into four groups, each group containing seven rats represent as follows: group 1 determined as control; group 2 determined as colitis-untreated; group 3 determined as colitis rats administered silymarin (50 mg/kg) and group 4 determined as colitis rats administered silymarin (100 mg/kg). Doses of 50 mg/kg and 100 mg/kg silymarin decreased tissue levels of malondialdehyde (MDA), cathepsin L and cathepsin B and activity of myleperoxidase (MPO) enzyme with respect to the colitis group (p<0.05). Based on the results of the study, it can be said that silymarin can be used as an effective treatment agent in inflammatory bowel diseases.

Open Access Original Research Article

Protein Uptake and Utilization of Moringa olifera Leaves and Seeds Fed to Rats as a Dietary Supplement

N. N. Umerah, A. I. Asouzu, J. I. Okoye

Asian Journal of Biochemistry, Genetics and Molecular Biology, Page 1-10
DOI: 10.9734/ajbgmb/2019/v2i430073

Background/Objective: Protein malnutrition is detrimental at any point in life. Protein malnutrition prenatally has also been shown to have significant lifelong effects. The objective of this study was to evaluate the protein uptake and utilization of Moringa olifera leaves and seeds fed to rats as a dietary supplement.

Materials and Methods: The leaves and seeds were harvested from the forest. The leaves were washed, drained and divided into three portions. The first portion was processed raw, the sun and shade dried samples were the second and third portion. The seeds were cracked and divided into six portions. The other five portions were fermented for 24, 48, 72, 96 and 120 h respectively. The blends of the shade dried leaves (SHDL) and 69 h fermented seed (FMS) provided 10% protein for two groups of rats. The diets were fed to twenty (20) adult albino rats in a 12-day N and mineral balance study. Casein was used as control. The result generated were statistically analyzed using SPSS version 22.

Results: The result showed that over 70% of the nitrogen consumed by all the two groups of rats fed the test diets were absorbed and retained. Among the test diet group, the FMS diet had the highest absorbed nitrogen (0.94 g), retained nitrogen (0.55 g), biological value (76%) and net protein utilization (52%) which was significantly different from the other groups. The SHDL had the most digested nitrogen (94%), highest maintenance food intake (74.09%) and weight gain (34.28%) that were comparable with the reference protein casein at (p>0.05). The FMS group had the highest liver, small intestine and lung weight.  The mineral metabolism of all the rats fed the test diets were absorbed and retained.

Conclusion: Generally, both the leaves and the seeds of Moringa olifera have good nutrient profile which could promote growth, they should be consumed to diversify diet and reduced micronutrient deficiency.