Identification of novel laccase from cyanobacterium Microcystis flos-aquae and enhanced azo dye bioremediation potential.

Textile industries discharge up to 280,000 tons of dye waste annually, resulting in global pollution and health risks. In Nigeria and other African countries, persistent dyes threaten aquatic life and human health. This study introduces a cost-effective, enzyme-mediated bioremediation alternative using a novel laccase from the cyanobacteriumMicrocystis flos-aquae. This purified enzyme yielded 0.55 % (w/w)with significant activity at 40 °C and pH 4.00. Kinetic studies showed the dependence of M. flos-aquae laccase on Cu2+and its inhibition by EDTA and Fe2+. The efficacy of the enzyme was demonstrated through rapid decolorization of the azo dye Cibacron Brilliant Blue over a wide temperature and pH range. As this enzyme effectively decolorizes dyes across a broad temperature and pH range, it offers a promising solution for bioremediation of textile effluents.

Toxicological analyses of the venoms of Nigerian vipers Echis ocellatus and Bitis arietans.

BACKGROUND: Among the medically important snakes in Nigeria, Echis ocellatus and Bitis arietans have the most lethal venom. These venoms were classified according to the presence of snake venom metalloproteinases (SVMPs), snake venom phospholipase A2 (PLA2s), and snake venom serine proteases (SVSPs). Toxicological analyzes were performed to understand the significance of different protein families in venoms. METHODS: Proteins were separated from venom using column chromatography. The skin and footpad of mice were used to determine hemorrhagic and edematogenic activities. Caprine blood plasma was used to test fibrinolytic activity in vitro. RESULTS: The results showed that, compared to the crude venom, the SVMP fraction induced hemorrhagic effects with a diameter of 26.00 ± 1.00 mm in E. ocellatus and 21.33 ± 1.52 mm in B. arietans. Both SVSP and SVMP had anticoagulant effects; however, the SVSP fraction had a stronger effect, with a longer anticoagulation time of 30.00 ± 3.00 min in E. ocellatus and 26.00 ± 2.00 min in B. arietans. These main venom toxins, SVMPs, SVSPs, and PLA2, were found to have edema-forming effects that were optimal at 2 h after envenomation. PLA2s had the highest edema-inducing activity, with onset 30 min after envenomation. CONCLUSIONS: Given the importance of SVMPs in altering the integrity of the membrane structure and impairing the blood coagulation system, an antivenom that can specifically neutralize its activity could inhibit the hemorrhage effects of the venoms.

Contribution of school meals to the recommended nutrient and energy intake of children enrolled in the National Homegrown School Feeding Program in Zaria, Nigeria.

BACKGROUND: A poor nutritional status in children results in reduced physical and mental health and poor academic performance. The National Homegrown School Feeding Program (NHSFP) was introduced in Nigeria in 2016 to ameliorate short-term hunger and improve the nutritional status of school-aged children (SAC). At least 33% of the recommended nutrient intake (RNI) for the enrolled students should be met by the school meals. However, to our knowledge, the contribution of school meals served through the NHSFP to the RNI of SAC in Zaria, Nigeria, remains to be explored. METHODS: We conducted a school-based cross-sectional study among 276 eligible SAC recruited from public primary schools in the Zaria Local Government Area. Portion sizes of the meals served through the NHSFP were determined using an electronic scale, meal samples were collected for nutrient analysis, and the average daily nutrient intake of the participants from the meals was calculated. The average daily intake of nutrients and energy of the participants was compared with the age- and sex-specific RNI to estimate the percentage contribution of the meals. RESULTS: The portion sizes recorded were 199.3 ± 20.6 g, 263.9 ± 11.5 g, 242.1 ± 16.8 g, 311 ± 17.3 g, and 160.3 ± 1.9 mL, respectively, for moi-moi, jollof rice and beans, bean porridge, jollof rice and boiled egg, and yoghurt. In addition, the meals contained moisture (30.13-66.11%), ash (0.73-7.08%), crude fat (9-32.61%), crude protein (7.25-24.5%), crude fiber (0-2.45%), and total carbohydrate (2.19-29.74%) with an energy content ranging from 183.6 to 330.57 kcal. Similarly, the meals contained calcium (82.58-711 mg), potassium (133-797 mg), sodium (340-1720 mg), iron (0.078-8.60 mg), zinc (1.84-13.4 mg), vitamin A (2.38-100.56 RAE), and vitamin C (0.04-1.57 mg) per 100 g of the school meals. The meals contributed 18.2-19.1%, 102.8-183.7%, 13.04-13.6%, and 26.1%-35.8% of the RNI for carbohydrates, proteins, fiber, and energy, respectively. Furthermore, they contributed 137-175%, 314.3-502.2%, 87.6-142.1%, 21.5-25.1%, 279.2-348.5%, 3.3-5.9%, 24.7-48.8%, and 3.3-5.9% of the RNI for iron, zinc, calcium, potassium, sodium, vitamin A, and vitamin C, respectively. CONCLUSION: The meals served through the NHSFP contributed at least 33% of the RNI for energy, protein, iron, calcium, sodium, vitamin A, and zinc. However, they could not meet the 33% of the RNI for carbohydrates, fiber, potassium, and vitamin C. Increasing the portion sizes and the diversity of the meals can address the suboptimal contribution of the meals to the RNI for carbohydrates, fiber, potassium, and vitamin C.

Kinetics and computational study of butyrylcholinesterase inhibition by methylrosmarinate: relevance to Alzheimer's disease treatment.

Butyrylcholinesterase (BChE) performs a significant function in Alzheimer's disease progression. Experimental studies have shown that the function of BChE in the attenuation of cholinergic neurotransmission is essentially altered in brains of advanced AD patients. Here, using the complimentary methods of enzyme kinetic studies, molecular modeling and protein-ligand interaction profiling, we sought to reveal the mechanistic and structural features of BChE-methyrosmarinate interactions. Molecular docking simulations revealed that methylrosmarinate dwelled well in the active centre of BChE, where it got involved in stabilizing non-covalent associations with myriad subsites. Enzyme kinetic experiments showed that the V m and K s values were 156.20 ± 3.11 U mg-1 protein and 0.13 ± 0.01 µM, respectively. The inhibition studies showed that methylrosmarinate apparently inhibited BChE in a linear mixed manner, with an IC 50 value of 10.31 µM and a K i value of 3.73 ± 1.52 µM. Taken together, the extremely reduced K i value and the increased number of BChE-methylrosmarinate interactions presuppose that methylrosmarinate is a good inhibitor of BChE, despite the fact that the mechanism for the effect of BChE inhibition on several pathological conditions in vivo remains unexplored.

Variations in the Serum Sialic Acid Profiles of Malaria Patients in Zaria, Nigeria: A Cross-Sectional Study.

PURPOSE: Understanding some variations in specialized molecules during malaria could facilitate adequate monitoring of patients and reduce the fatalities caused by the disease. The present study reports changes in the levels of free serum sialic acid (FSSA) among Plasmodium-infected individuals in Zaria, Nigeria, in a cross-sectional study with 170 individuals. METHODS: The FSSA and total sialic acid (TSA) in the blood were determined using the thiobarbituric acid method and the white blood cells (WBC) count, haemoglobin concentration and packed cell volumes were assessed using an automated haematological analyser. RESULTS: The results showed that, in the patients aged > 5 years the level of TSA was significantly elevated (P < 0.05) compared to apparently healthy age-matched controls whereas TSA was slightly lower in patients aged < 5 years compared to controls. The ratio of FSSA to TSA was not different between patients aged > 5 years compared to their age-matched controls whereas FSSA/TSA was significantly elevated (P < 0.05) in patients aged < 5 years compared to their aged-matched controls. The level of FSSA/TSA in the patients aged < 5 years was not correlated with parasite density, white blood cell count, haemoglobin concentration or packed cell volume. CONCLUSION: We concluded that, metabolism and/or physiology of serum sialo-glycoconjugates is affected by malaria and FSSA is mainly elevated in children < 5 years of age but not among older patients suggesting the possible usefulness of FSSA in the analysis of uncomplicated malaria in under five children.

Venom proteomic analysis of medically important Nigerian viper Echis ocellatus and Bitis arietans snake species.

Snakebite envenoming remains a neglected tropical disease which poses severe health hazard, especially for the rural inhabitants in Africa. In Nigeria, vipers are responsible for the highest number of deaths. Hydrophilic interaction liquid chromatography coupled with LC-MS/MS was used to analyze the crude venoms of Echis ocellatus (Carpet viper) and Bitis arietans (Puff adder) in order to understand their venom proteomic identities. Results obtained revealed that gel-free proteomic analysis of the crude venoms led to the identification of 85 and 79 proteins, respectively. Seventy-eight (78) proteins were common between the two snake species with a 91.8% similarity score. The identified proteins belong to 18 protein families in E. ocellatus and 14 protein families in B. arietans. Serine proteases (22.31%) and metalloproteinases (21.06%) were the dominant proteins in the venom of B. arietans; while metalloproteinases (34.84%), phospholipase A2s (21.19%) and serine proteases (15.50%) represent the major toxins in the E. ocellatus venom. Other protein families such as three-finger toxins and cysteine-rich venom proteins were detected in low proportions. This study provides an insight into the venom proteomic analysis of the two Nigerian viper species, which could be useful in identifying the toxin families to be neutralized in case of envenomation.

Interaction of SARS-CoV-2 spike protein with angiotensin converting enzyme inhibitors and selected compounds from the chemical entities of biological interest.

BACKGROUND: Recent COVID-19 outbreak has prompted the search of novel therapeutic agents to treat the disease. The initial step of the infection involves the binding of the virus through the viral spike protein with the host angiotensin converting enzyme 2 (ACE2). In this study, the interaction of some ACE or ACE2 inhibitors and their analogues as well as selected compounds with the viral spike protein as a strategy to hinder viral-ACE2 interaction were investigated. SARS-CoV-2 spike protein as well as the ligands were retrieved from protein databank and ChEBI database respectively. The molecules were prepared before initiating the virtual screening using PyRx software. Discovery studio was used to further visualize the binding interactions between the compounds and the protein. RESULTS: The ACE inhibitors and their analogues fosinopril (1-), fosinopril and moexipril have the best binding affinity to the protein with binding energies < - 7.0 kcal/mol while non-flavonoid stilben-4-ol binds with free binding energy of - 7.1 kcal/mol. Others compounds which belong to either the flavonoids, terpenes and alkaloid classes also have binding energies < - 7.0 kcal/mol. Such high binding energies were enhanced via hydrogen bond (h-bond) interactions in addition to other interactions observed between the compounds and the amino acid residues of the protein. CONCLUSIONS: The ACE inhibitors and their analogues as well as the selected compounds could serve as inhibitors of the spike protein as well as lead in drug discovery processes to target the SARS-CoV-2 virus.

Proteomic analysis of three medically important Nigerian Naja (Naja haje, Naja katiensis and Naja nigricollis) snake venoms.

Proteomics technologies enable a comprehensive study of complex proteins and their functions. The venom proteomes of three medically important Nigerian Elapidae snakes Naja haje, Naja katiensis and Naja nigricollis was studied using HILIC coupled with LC-MS/MS analysis. Results revealed a total of 57, 55, and 46 proteins in the venoms of N. haje, N. katiensis, and N. nigricollis, respectively, with molecular mass ranging between 5 and 185  kDa. These snakes have 38 common proteins in addition to 3 uncommon proteins: actiflagelin, cathelicidin, and cystatin identified in their venoms. The identified proteins belonged to 14 protein families in N. haje and N. katiensis, and 12 protein families in N. nigricollis. Of the total venom proteins, 3FTx was the most abundant protein family, constituting 52% in N. haje and N. katiensis, and 41% in N. nigricollis, followed by PLA2, constituting 37% in N. nigricollis, 26% in N. haje, and 24% in N. katiensis. Other protein families, including LAAO, CRISPs, VEGF, PLB, CVF, SVMP, SVH, AMP, PI, Globin, Actin, and C-type lectins, were also detected, although, at very low abundances. Quantification of the relative abundance of each protein revealed that alpha and beta fibrinogenase and PLA2, which constituted 18-26% of the total proteome, were the most abundant. The 3 uncommon proteins have no known function in snake venom. However, actiflagelin activates sperm motility; cystatin inhibits angiogenesis, while cathelicidin exerts antimicrobial effects. The three Nigerian Naja genus proteomes displayed 70% similarity in composition, which suggests the possibility of formulating antivenom that may cross-neutralise the venoms of cobra species found in Nigeria. These data provide insights into clinically relevant peptides/proteins present in the venoms of these snakes. Data are available via ProteomeXchange with identifier PXD024627.

Vitex doniana Leaves Extract Ameliorates Alterations Associated with 7, 12-Dimethyl Benz[a]Anthracene-Induced Mammary Damage in Female Wistar Rats.

Vitex doniana leaves are used traditionally in West Africa for the treatment of swellings and cancer. We investigated if Vitex doniana leaves extract could ameliorate 7, 12-dimethylbenz[a]anthracene (DMBA)-induced mammary damage. Female Wistar rats aged 52 ± 2 day were administered 80 mg/kg DMBA. After monitoring for 150 day, rats were administered 0, 50, 100, 200 mg/kg Vitex doniana and 20 mg/kg Tamoxifen for 14 day. Serum estrogen receptor-α, IL-1ß and TNF -α levels were determined using ELISA kits. Oxidative stress markers in mammary tissue homogenates were determined using standard spectrophotometric methods. Histopathological examination was done using hematoxylin and eosin staining and cyclooxygenase-2 (COX-2) expression using immunohistochemistry. Liquid chromatography-mass spectrometry was used to determine components present in the extract. Although tumors were not observed, significantly (p < 0.05) lower estrogen receptor-α, malondialdehyde, IL-1ß and TNF -α levels, significantly (p < 0.05) higher glutathione and catalase activity, attenuation of malignant epithelial hyperplasia and mild COX-2 expression were observed in rats administered Vitex doniana when compared to DMBA-induced untreated control. Liquid chromatography-mass spectrometry analysis of the V. doniana extract revealed the presence of 4,5-dihydroxy-7-methoxy-6-methylflavone and vanillylamine, which are compounds with reported antioxidant and anti-inflammatory effects. Collectively, treatment with Vitex doniana ameliorated some derangement observed in DMBA-induced rats.

Depletion of cholesterol could be associated with modulation of progesterone but not other sex hormone levels during Plasmodium falciparum infection in humans: a cross-sectional study from Zaria, Nigeria.

In order for Plasmodium falciparum to grow and survive in its host, membrane biogenesis, fueled by host cholesterol, is essential for these processes. Consistent with this essential role, more insights into the cholesterol pathway would enhance the current understanding of the pathophysiology of malaria infection. To explore its broader potential, we conducted a cross-sectional study and assayed for the serum levels of cholesterol, vitamin D, progesterone, testosterone, estradiol and bile acid in both P. falciparum-infected patients and apparently healthy sex-matched participants. Our results revealed that the levels of cholesterol, vitamin D, progesterone, testosterone and estradiol in P. falciparum-infected patients were significantly (p < 0.05) lower compared to those in control groups whereas the level of bile acid in P. falciparum-infected patients was significantly (p < 0.05) higher compared to that in control groups. Additionally, cholesterol and the metabolic products with the exception of bile acid had a significant (p < 0.05) association with the parasite density in P. falciparum-infected patients with moderate and high P. falciparum infections. Furthermore, all the metabolic products of cholesterol had an insignificant (p > 0.05) association with the cholesterol in P. falciparum-infected patients with the exception of progesterone which showed a significant (p < 0.05) association with cholesterol in the malaria-infected female patients. Data from the present study demonstrated that progesterone depletion in P. falciparum-infected female patients could be a consequence of P. falciparum-induced decrease in cholesterol.

Inhibition of Trypanosoma evansi Protein-Tyrosine Phosphatase by Myristic Acid Analogues Isolated from Khaya senegalensis and Tamarindus indica.

BACKGROUND: Trypanosome infections still pose severe health and economic consequences, especially in the endemic regions of Sub-Saharan Africa. Trypanosome differentiation to the procyclic forms which lack the immune evasion mechanisms for survival in the bloodstream is prevented by tyrosine dephosphorylation which is catalyzed by protein-tyrosine phosphatase; thereby promoting survival of the parasites in the host. Inhibition of Protein-tyrosine phosphatase is a strategic therapeutic target that could attenuate trypanosomiasis. This study investigated the in vitro inhibitory effect of stem bark extracts of Khaya senegalensis and Tamarindus indica on the enzymatic activity of protein-tyrosine phosphatase. METHODS: All determinations were carried out following standard procedures for analytical experiments. The analogues of myristic acid that inhibited the enzymatic activity of protein-tyrosine phosphatase were isolated by bioassay-guided fractionation of stem bark extracts of Khaya senegalensis and Tamarindus indica. RESULTS: Analogues of myristic acid proved to be potent inhibitors of protein-tyrosine phosphatase. Double reciprocal (Lineweaver-Burk) plots of the initial velocity data indicated non-competitive inhibition with Ki of 0.67 mg/mL for Khaya senegalensis and 2.17 mg/mL for Tamarindus indica. The kinetic parameters for the cleavage of para-nitrophenylphosphate by the enzyme showed a KM of 3.44 mM and Vmax of 0.19 µmol/min. Sodium orthovanadate, the enzymes' specific inhibitor, inhibited the enzyme competitively with Ki of 0.20 mg/mL. Gas chromatography-mass spectrometry analysis of the stem bark bioactive fractions of Khaya senegalensis and Tamarindus indica revealed the presence of myristic acid analogues. CONCLUSION: Analogues of myristic acid are potent inhibitors of protein-tyrosine phosphatase that could be developed as trypanocide to inhibit the enzymatic activity of protein-tyrosine phosphatase in order to prevent transmission of trypanosomes.

Oxidative Stress and Carcinogenesis: Potential of Phytochemicals in Breast Cancer Therapy.

Breast cancer remains a burden in both developed and developing countries, with higher mortality in developing countries. Attempts to eradicate cancer have not been successful despite the progress made in the development of more novel chemotherapeutic drugs. Reactive-oxygen-species-mediated oxidative stress is known to play a role in breast cancer pathogenesis via genetic and epigenetic modifications, resulting in uncontrolled cell proliferation. Phytochemicals could provide leads for the development of alternative therapeutic agents due to their antioxidant activity, as well as their ability to induce apoptosis in cancer cells. However, most of the studies carried out using in vitro models do not continue with further studies in estrogen-receptor-positive in vivo breast cancer models, or fail to examine the possible biochemical mechanisms of phytochemical-based amelioration. This review examines oxidative-stress-mediated carcinogenesis and the potential of phytochemicals as anticancer agents.

Agricultural residues for cellulolytic enzyme production by Aspergillus niger: effects of pretreatment.

Different agricultural residues were considered in this study for their ability to support cellulolytic enzyme production by Aspergillus niger. A total of eleven agricultural residues including finger millet hulls, sorghum hulls, soybean hulls, groundnut husk, banana peels, corn stalk, cassava peels, sugarcane bagasse, saw dust, rice straw and sheanut cake were subjected to three pretreatment (acid, alkali and oxidative) methods. All the residues supported the growth and production of cellulases by A. niger after 96 h of incubation. Maximum cellulase production was found in alkali-treated soybean hulls with CMCase, FPase and ß-glucosidase yields of 9.91 ± 0.04, 6.20 ± 0.13 and 5.69 ± 0.29 U/g, respectively. Further studies in assessing the potential of soybean hulls are being considered to optimize the medium composition and process parameters for enhanced cellulase production.

Hepatocurative potential of Vitex doniana root bark, stem bark and leaves extracts against CCl4-induced liver damage in rats

<p><b>OBJECTIVE</b>To evaluate the hepatocurative effects of aqueous root bark, stem bark and leaves of Vitex doniana in carbon tetrachloride (CCl4) induced liver damage and non induced liver damage albino rats.</p><p><b>METHODS</b>A total of 60 albino rats (36 induced liver damage and 24 non induced liver damage) were assigned into liver damage and non liver damage groups of 6 rats in a group. The animals in the CCl4 induced liver damage groups, were induced by intraperitoneal injection with a single dose of CCl4 (1 mL/kg body weight) as a 1:1(v/v) solution in olive oil and were fasted for 36 h before the subsequent treatment with aqueous root bark, stem bark and leaves extracts of Vitex doniana and vitamin E as standard drug (100 mg/kg body weight per day) for 21 d, while the animals in the non induced groups were only treated with the daily oral administration of these extracts at the same dose. The administration of CCl4 was done once a week for a period of 3 weeks.</p><p><b>RESULTS</b>There was significant (P<0.05) increase in concentration of all liver marker enzymes, alanine aminotransferase, aspartate aminotransferase and alkaline aminotransferase (ALT, AST and ALP) and significant (P<0.05) decrease in albumin in the CCl4 induced liver damage control when compared to the normal control. The extracts caused a significant (P<0.05) reduction in the serum activities of liver marker enzymes (ALT, AST and ALP) and a significant (P<0.05) increase in albumin of all the induced treated groups. Only stem bark extract and vitamin E significantly (P<0.05) increased total protein. All the extracts significantly (P<0.05) lowered serum creatinine whereas only root bark extract significantly (P<0.05) lowered serum level of urea in the rats with CCl4 induced liver damage.</p><p><b>CONCLUSION</b>Hepatocurative study shows that all the plant parts (root bark, stem bark and leaves) possess significant hepatocurative properties among other therapeutic values justifying their use in folklore medicine.</p>

Hepatocurative potential of Vitex doniana root bark, stem bark and leaves extracts against CCl

Objective: To evaluate the hepatocurative effects of aqueous root bark, stem bark and leaves of Vitex doniana in carbon tetrachloride (CCl