International Journal of Biochemistry Research & Review <p style="text-align: justify;"><strong>International Journal of Biochemistry Research &amp; Review (ISSN: 2231-086X)</strong> publishes original research papers, review articles and short communications on all areas of Biochemistry.&nbsp;This is a quality controlled,&nbsp;OPEN&nbsp;peer reviewed,&nbsp;open access INTERNATIONAL journal.</p> SCIENCEDOMAIN international en-US International Journal of Biochemistry Research & Review 2231-086X The Effect of Ethanolic Extract of Cannabis sativa Leaves from Nigeria on the Antioxidants Markers in Albino Wistar Rats <p><strong>Background:</strong> <em>Cannabis sativa</em> is an annual herbaceous plant in the Cannabis genus. The cannabis plant is widely regarded as a potent psychoactive, medicinal plant. Reportedly used for recreation and as intoxicant. The medical uses of the plant include effective control and management of chronic health problems such as HIV/AIDs, cancer, cachexia, nausea and vomiting, asthma and hypertension. <em>C. sativa</em> is known to possess antioxidative properties. This study therefore investigated the effects of <em>C. sativa</em> on antioxidant concentrations in albino Wistar rats.</p> <p><strong>Materials and Methods: </strong>Thirty (30) rats used for this study were divided into three groups of 10 rats each. Group 1 received distilled water and served as control. Group 2 received <em>C. sativa</em> extract (100 ml/Kg body weight) by gavage and served as low dose group. Group 3 received <em>C. sativa</em> (250 ml/Kg body weight) by gavage and served as high dose group once daily for 28 days.</p> <p><strong>Results: </strong>Catalase (CAT) concentration was significantly lower (P&lt;0.05) in the low dose group as compared with control. In the high dose group, CAT concentration was significantly lower (P&lt;0.05) when compared with the low and control groups respectively. Glutathione peroxidase (GPx) and Superoxide dismutase (SOD) concentrations were significantly higher (P&lt;0.05) in the low dose group as compared with their respective control. GPx and SOD concentrations were significantly higher (P&lt;0.05) in the high dose groups as compared to low dose and control groups respectively.</p> <p><strong>Conclusion</strong>: CAT concentration decreased dose-dependently, while GPx and SOD concentrations increased dose-dependently among treated groups. Treatment with <em>C. sativa</em> revealed a paradoxical effect on CAT concentration with respect to GPx and SOD concentrations. Therefore, oral ingestion of ethanolic extract of <em>C. sativa</em> may not have significant effect on the body’s antioxidant stores due to the balance created for CAT deficiency by increased GPx and SOD concentrations.</p> E. B. Umoren I. Wopara O. G. Adebayo U. A. Ezike A. O. Obembe ##submission.copyrightStatement## 2020-11-26 2020-11-26 58 65 10.9734/ijbcrr/2020/v29i930225 Evaluation of Formulated Anti-dermatophyte Creams from Ethanol Extract of Mitracarpus villosus Leaves <p><em>Microsporium, Trichophyton</em>, and <em>Epidermophyton</em> are asexual fungi usually called dermatophytes. They are known to cause a kind of skin diseases called dermatophytosis. There have been reported cases of resistance by the dermatophytes with prolonged usage of synthetic antifungals on the skin. The aim of this study was to formulate herbal antifungal cream containing extract of <em>Mitracarpus villosus </em>as an anti-dermatophytic preparation and evaluate its physicochemical properties, stability and efficacy of the product. The formulated creams containing 0.5, 1 and 2% w/w of extract were subjected to stability tests using temperature variation method at -10, 4, 30, 37 and 45<sup>o</sup>C. Freeze-thaw test, Centrifuge test, pH and exposure to UV light test were also carried out using standard method. Efficacy of the cream formulations were determined using Wistar rats as experimental animals.</p> <p>The percentage yield of the extract was (2.1%). Percentage ethanol phytochemical composition indicated that for alkaloid it is 1.06±0.04%, saponins (0.96±0.07%), flavonoids (0.06±0.02%) and tannins (0.04±0.01%). The emulsion produced was an oil-in-water emulsion and had a white colour with pH of 7.02, spread of emulsion, rubbing-in effect and stability to centrifugation was very high. The antifungal results showed the activity against the dermatophytes to be in the increasing order <em>Epidermophyton floccosum </em>(9 mm) &lt;<em>Microsporum audounii </em>(12 mm) <em>&lt;</em>Trichophyton<em> mentagrophtes </em>(13 mm) &lt;<em>M. furfur </em>(12 mm).Temperature stability and Centrifuge testing indicated that the formulations were stable. Light testing indicated no change in the colour of the products. Animal studies evaluation of the formulations of the cream indicated that their efficacy against the dermatophytes is concentration dependent and is in the increasing order<em> M. audounii </em>(34 µm) = <em>E. floccosum </em>(34 µm) &lt;<em>T. mentagrophyte </em>(35 µm) &lt;<em>M. furfur </em>(45 µm) which shows that 2%<em> Mitracarpus villosus</em> cream was statistically significant (P&lt;0.05) against all the test microorganisms.</p> A. B. Fawehinmi F. O. Oyedeji ##submission.copyrightStatement## 2020-10-29 2020-10-29 1 12 10.9734/ijbcrr/2020/v29i930219 A Comparative Study between Plant and Callus Extracts of Abutilon indicum (L.) Sweet: Antioxidant, Antibacterial, Antidiabetic and Anti-Proliferative Activity <p><em>Abutilon indicum</em> is consider to be used in the traditional system of medicine. It is found in tropical and subtropical regions of the world. It is used to treat various diseases. This plant does not cause any side effects to humans. As the plant has wide variety of medicinal properties, the present study aimed to comparative between plant and callus extract of <em>Abutilon indicum </em>(L.) sweet for antioxidant, antibacterial, antidiabetic and anti- proliferative activity. The highest percentage of callus induction (89.50%) and callus weight (1.26 g) was observed in T<sub>5</sub> (MS + 2, 4-D (2.5 mg/l) + BAP (2 mg/l) and T<sub>8</sub> [IBA (4 mg/l)] respectively. Phytochemical analysis of aqueous and ethyl acetate extracts of <em>A. indicum in vivo plant and in vitro grown callus</em> showed the presence of alkaloids, flavonoids, phenols, carbohydrates, glycosides, protein, terpenoids, saponins, tannins and coumarin. The total phenolic content was high in aqueous extract of callus (30.68 mg TAE/g). Maximum DPPH radical scavenging activity was found in aqueous extract of callus (86%) with IC<sub>50</sub> value of 68.49 µg/ml. FT-IR analysis of aqueous extract of <em>A. indicum </em>plant and callus showed the presence of characteristic stretching at 2930.28 and 2927.75 indicating the presence of C-H stretching respectively. GC-MS analysis revealed the presence of 17 compounds in ethyl acetate plant extract, whereas 7 compounds in ethyl acetate callus extract such as tetradecane, 1-chloro, Sulfurous acid 2-prophytridecyl ester and 1- ethyl-3-[2-(octadecylthio) ethyl] thiourea. The ethyl acetate extracts of callus and plant and was found to be effective against <em>Bacillus subtilis </em>(3.1 mm) and <em>Staphylococcus aureus </em>(2.9 mm)<em>.</em> Maximum α-amylase inhibitory activity was observed in aqueous callus extract (32.65%) with IC<sub>50</sub> value of 833.61 µg/ml. HeLa cell viability was found to be 26.8% and 21.8% in plant and callus extract respectively.</p> A. Ayisha Sireen J. Anbumalarmathi ##submission.copyrightStatement## 2020-11-04 2020-11-04 13 24 10.9734/ijbcrr/2020/v29i930220 Use of Trichoderma in Biological Control of Collar Rot of Soybean and Chickpea <p>An in vitro and field experiments for two consecutive years were conducted at Bangladesh Institute of Nuclear Agriculture, Mymensingh, aiming to investigate the efficacy of Trichoderma harzianum against Sclerotium rolfsii causing collar rot disease of soybean and chickpea. In in vitro the antagonistic activity of T. harzianum against S. rolfsii was observed through dual culture. In field experiment Trichoderma was applied as soil treatment and seed treatment. The percent inhibition of S. rolfsii induced by T. harzianum was found upto 78.9% in in vitro. The maximum reduction of collar rot disease incidence over control was 82.4% in soybean and 77.6% in chickpea which was recorded in the plot where T. harzianum was applied in the soil. The highest seed germination: 86.3% in soybean and 84.8% in chickpea, maximum fresh shoot weight: 94.5 g plant<sup>-1</sup> in soybean, 62.5 g plant<sup>-1</sup> in chickpea, maximum fresh root weight: 10.7 g plant<sup>-1</sup> in soybean, 9.3 g plant<sup>-1</sup> in chickpea and the highest yield: 2830 kg ha<sup>-1</sup> in soybean, 1836 kg ha<sup>-1</sup> in chickpea were obtained by the application of Trichoderma in soil. The study indicated that the tested isolate of T. harzianum had potential in controlling collar rot disease of soybean and chickpea. For the reduction of collar rot incidence application of T. harzianum in soil was found more effective than seed treatment.&nbsp;</p> Mahbuba Kaniz Hasna Md. Abul Kashem Farid Ahmed ##submission.copyrightStatement## 2020-11-07 2020-11-07 25 31 10.9734/ijbcrr/2020/v29i930221 The Effects of Garlic Extract on the Proximate Composition and Microbial Load of Hot and Cold-Smoked Clupea harangus (Atlantic Herring Fish) <p>The study was carried out to determine the effect of garlic extract on the proximate composition and microbial activity of cold and hot smoked <em>Clupea harangus (</em>Atlantic herring fish<em>)</em>. In the study, fresh atlantic herring was thawed, eviscerated, weighed, washed properly and folded. Garlic extract was prepared at 100 ml, 75 ml, 50 ml and control with no garlic extract, each treatments were prepared in duplicates and in three batches. The folded atlantic herring was dipped into the garlic extract at the different concentration, the first batch of treatments was oven dried for 40°C for 6 hours (cold smoked), the second batch was oven dried for 65°C for 12 hours (cold smoked) and the third batch was oven dried for 75°C for 18 hours (hot smoked). After oven drying, it was allowed to cool for 20 minutes, samples for microbial analysis was immediately taken in a foil paper and stored in the fridge for a month while the remaining sample was immediately used for proximate analysis. At all hours of drying period, the moisture content decreases and protein content increases with increasing concentration of garlic extract, the lipid content was highest at 75 ml garlic extract and the ash content was highest at 50 ml garlic extract. Total bacteria count decreases with increasing concentration of garlic, increasing temperature and increasing hour of drying. No fungi growth was recorded at all hours of drying period.</p> Sarah Tosin Fashagba Badiu Akinola Akinbode Sherifdeen Olamilekan Babalola ##submission.copyrightStatement## 2020-11-11 2020-11-11 32 39 10.9734/ijbcrr/2020/v29i930222 Investigating the Modulatory Effect of Methanol Extract of Daniellia oliveri (ROLFE) Leaves on Mitochondrial Membrane Permeability Transition (MPT) Pore <p><strong>Background:</strong> Mitochondrial-mediated cell death begins with opening of mitochondrial membrane permeability transition (mPT) pore and medicinal plants contain phytochemicals that modulate the mPT pore.</p> <p><strong>Hypothesis and Purpose:</strong> We investigated the modulatory effects of crude methanol extract of Daniellia oliveri leaves (CMDO) on mPT pore <em>in</em> <em>vitro</em>.</p> <p><strong>Study Design and Methods:</strong> Phytochemical screening and antioxidant activities of crude methanol extract of <em>Daniellia</em> <em>oliveri</em> leaves (CMDO) were evaluated according to standard procedures. CMDO was partitioned into chloroform fraction (CFDO), ethyl acetate fraction (EFDO) and methanol fraction (MFDO) by Vacuum liquid chromatography (VLC). Effects of CMDO, CFDO, EFDO and MFDO on mPT pore were assessed by spectrophotometry. Effects of the most potent fraction on mitochondrial ATPase, Fe-induced lipid peroxidation and cytochrome c release were assessed by spectrophotometry. CMDO was subjected to GC-MS analysis to identify the bioactive compounds present.</p> <p><strong>Results:</strong> CMDO contains phytochemicals and showed appreciable total flavonoid content (0.483±0.02 QE mg/100g), total phenolic content (0.886±0.12 GAE mg/100g), total antioxidant capacity (0.039±0.001 AE mg/100 g), ferric antioxidant reducing power (IC50=350 µg/ml) and 2, 2-diphenyl-1 picrylhydrazyl (DPPH) radical scavenging activity (IC50=166 µg/ml). The maximum induction of mPT pore opening in the absence and presence of calcium, respectively, were as follows: CMDO (10.11 folds, 5.18 folds), CFDO (19.9 folds, 16.3 folds), EFDO (7.5 folds, 23.2 folds), MFDO (22.2 folds, 31.3 folds). The most potent mPT pore-opening fraction (MFDO) enhanced mitochondrial ATPase activity, inhibited Fe-induced lipid peroxidation and caused cytochrome c release. GC-MS analysis of CMDO revealed the presence of bioactive compounds including methyl propanamide, Dibutyl phthalate, saturated and unsaturated fatty acids.</p> <p><strong>Conclusion:</strong> Methanol fraction (MFDO) of CMDO most potently induced mPT pore opening via enhancement of mitochondrial ATPase activity, which was substantiated by the release of cytochrome c (<em>in</em> <em>vitro</em>). This includes MFDO as a candidate pharmacologic remedy for diseases associated with insufficient apoptosis.</p> Jonah Achem Cosmos Ifeanyi Onyiba Mobolaji T. Akinwole Jemimah M. Malgwi Omosola L. Bolarin Olufunso O. Olorunsogo ##submission.copyrightStatement## 2020-11-12 2020-11-12 40 51 10.9734/ijbcrr/2020/v29i930223 Serum Copper and Serum Zinc in Preeclampsia: Cause or Effect? <p><strong>Background:</strong> Preeclampsia is multisystem disorder. Despite its prevalence and severity, the pathophysiology of this multisystem disorder is poorly understood. In concern regarding the increasing number of preeclamptic cases and lack of data about the levels of trace elements in preeclampsia, a case-control study was conducted with aim to determine the trace elements like serum total copper and serum total zinc in preeclampsia.</p> <p><strong>Aims:</strong> To estimate alterations in serum copper and serum zinc in preeclampsia and to compare them with normal pregnant women.</p> <p><strong>Study Design:</strong> This is a case control study, carried out in the Department of Biochemistry, MIMER Medical College, Talegaon Dabhade, Pune.</p> <p><strong>Methods: </strong>The present study consisted of 120 study participants. These were divided into two groups. Group I - normal pregnant women as control (n=60) and Group II - preeclamptic group (n=60). The serum levels of copper and zinc were determined by inductively coupled plasma atomic emission spectrometry (ICP- AES) technique at IIT Mumbai.</p> <p><strong>Results: </strong>Analysis revealed that mean values of total serum copper and total serum zinc were 196.20 ± 25.9 and 77.15 ± 14.5 (µg/dl) respectively in control group. In preeclamptic group, the mean values of copper and zinc were 213.13± 38.6 and 76.23 ± 13.13 (µg/dl) respectively. Copper was significantly increased in preeclamptic group, while non-significant reduction in levels of zinc levels was observed when compared to control group.</p> <p><strong>Conclusion: </strong>In the present study, significantly high serum copper was observed in preeclamptic patients. Presence of high copper levels may be related factor in the etiopathogenesis of preeclampsia. Estimation of trace elements like copper and zinc may help clinicians in early diagnosis and minimizing or delaying complications of preeclampsia, hence preventing harm to both mother &amp; fetus.</p> Anjum A. K. Sayyed Alka N. Sontakke ##submission.copyrightStatement## 2020-11-20 2020-11-20 52 57 10.9734/ijbcrr/2020/v29i930224 Manganese Inhibits Indomethacin-Induced Hepatorenal Oxidative Stress in Wistar Rats <p><strong>Aim:</strong> Manganese (Mn) is an essential trace element in many cellular processes. However, there is dearth of literature on its influence on indomethacin-induced hepatorenal damage. Therefore, this study was conducted to investigate the effect of manganese on indomethacin-induced hepatorenal damage in rats.</p> <p><strong>Methods: </strong>Rats were divided into four groups of eight rats consisting of control group, indomethacin (IND) alone (20 mg/kg), Mn alone (10 mg/kg) and co-treated group that were treated orally for 14 consecutive days. Twenty four hours after treatment, under pentobarbital anesthesia, blood was collected and liver was excised to prepare homogenate and histology staining. Liver and kidney function tests aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), gamma glutamyl transferase (GGT), lactate dehydrogenase (LDH), malate dehydrogenase (MDH), glutamine dehydrogenase (GLDH), sorbitol dehydrogenase (SDH), glucose-6-phosphate dehydrogenase (G6PD), bilirubin (BIL), urea, creatinine, cholesterol (CHOL), triglycerides (TG), low and high density lipoprotein (LDL and HDL), electrolytes and oxidative stress superoxide dismutase (SOD), catalase (CAT), glutathione (GSH) and lipid peroxidation (LPO) biomarkers were assessed.</p> <p><strong>Results: </strong>The results showed that indomethacin caused hepatorenal damage in rats manifested with increase in serum hepatic and renal function biomarkers. But co-administration of IND with Mn significantly (p &lt; 0.05) decreased the level of hepatorenal biomarkers. Additionally, co-administration of IND with Mn improved the antioxidant status with concomitant reduction of LPO and restored the integrity of the liver and kidney histologically.</p> <p><strong>Conclusion:</strong> The results of this study emphasize that co-administration of IND with Mn to rats alleviated IND-induced hepatorenal toxicities and oxidative stress in rats.</p> Tijani Stephanie Abiola Olori Ogaraya David Farombi Ebenezer Olatunde ##submission.copyrightStatement## 2020-11-27 2020-11-27 79 90 10.9734/ijbcrr/2020/v29i930227 The Effect of Aqueous Extract of Vernonia Amygdalina (Bitter Leaf) on Antioxidants and the Liver of Rabbits on High Cholesterol Diet <p><strong>Aims</strong><strong>:</strong> To determine the effect of<em> V.</em> <em>amygdalina </em>on the levels of antioxidants and the liver of rabbits fed on a atherogenic diet.</p> <p><strong>Study Design:</strong> Prospective experimental study.</p> <p><strong>Place and Duration of study:</strong> Study lasted 14 weeks at Department of Anatomy, University of Ibadan, Nigeria.</p> <p><strong>Methodol</strong><strong>o</strong><strong>gy</strong><strong>:</strong> Eighteen male rabbits were randomly divided into three groups comprising control, high cholesterol diet and high cholesterol diet plus 200mg/kg of aqueous extract of <em>V.</em> <em>amygdalina</em>. The animals were fed a normal animal chow, or a diet supplemented by 0.5% cholesterol and 1% methionine for twelve weeks. Subsequently, serum was obtained for liver function tests. Samples of the liver of the animals were obtained for antioxidant tests and histology.</p> <p><strong>Results:</strong> Aspartate transaminase was significantly elevated in atherogenic diet only group compared to animals on normal diet. There was also no statistically significant difference across the three groups in the alanine transaminase values. Superoxide dismutase were significantly reduced in animals that were administered extract. Malondialdehyde was not statistically different across the three groups.</p> <p>The histology of the liver of the animals on high cholesterol diet (with and without extract administration) revealed similar microscopic presence of fatty infiltration of the liver.</p> <p><strong>Conclusion:</strong> The use of high cholesterol diet for a duration of twelve weeks in rabbits resulted in significant increased relative liver weight, aspartate transaminase, superoxide dismutase and hepatic microvesicular steatosis. Furthermore, the use of 200mg/kg of aqueous extract of <em>V. amygdalina</em> had ameliorative effect on the level of superoxide dismutase. It did not have a measurable effect on the malondialdehyde, liver enzymes and liver histology.</p> O. Abdulmalik O. O. Oladapo ##submission.copyrightStatement## 2020-11-28 2020-11-28 91 100 10.9734/ijbcrr/2020/v29i930228 Low Cost Tissue Culture Technologies in Vegetables: A Review <p>The demand of vegetable crops is increasing day by day due to changes in consumption patterns, so the need of the hour is to develop technologies that enhance the vegetable production at a rapid rate. Plant Tissue culture is one such remarkable biotechnological tool that has its application in vegetable propagation and improvement, disease elimination, herbicide resistance, salinity tolerance, incorporation of high nutrient content, genetically improved plants and conservation of endangered plant species and in the near future usage of this technology is going to increase further manifold. It is used for production of disease free quality planting material and development of varieties through direct regeneration, anther/ovule culture, somatic embryogenesis etc. or for creation of new variation (organogenesis <em>via</em> callus formation, soma-clonal variation and <em>in vitro</em> mutagenesis). In spite of being a very important and viable non-conventional biotechnological tool, high cost of production of seedlings <em>in vitro</em> remains a major impediment in popularization of this technology. High cost of producing seedlings is due to availability of limited resources, high recurrent costs of consumables for media and lack of awareness, which limits its application only to a few institutions and rich farmers especially in developing countries. Therefore, in order to make this technology a successful and viable option for the farmers, future thrust must be on cost reduction of <em>in vitro</em> seedlings. The components of tissue culture technology such as culture media components, glassware, lighting and water for media preparation can be replaced with low cost alternatives to reduce the overall cost of tissue culture. The usage of alternatives for gelling agent’s like isabgol (potato, tomato, cassava, turmeric, ginger), sago (potato, tomato, turmeric, ginger) cassava starch (potato, cassava, sweet potato) barley starch, phytagel etc. and for carbon sources like table sugar (potato, turmeric, ginger), jaggery, sugarcane juice, cube sugar (bittergord), brown sugar etc have already been documented worldwide. The present paper reviews the work done by researchers around the globe in developing various low cost alternative technologies with focus on vegetable crops.</p> Reshav Naik Anil Bhushan R. K. Gupta Anamika Walia Anshika Gaur ##submission.copyrightStatement## 2020-11-27 2020-11-27 66 78 10.9734/ijbcrr/2020/v29i930226