Open Access Original Research Article

Antidermatophyte Activity of Ethanol Extract and Stability of Creams Formulated from Cassia alata Linn

F. O. Oyedeji, A. B. Fawehinmi, Hassan Lawal

International Journal of Biochemistry Research & Review, Page 1-8
DOI: 10.9734/ijbcrr/2020/v29i1030234

There are scientific reports of medicinal plants having curative properties. These properties may be due to the presence of various phytochemicals in the plants. Cassia alata is a specie in the Fabaceae family. The purpose of this study is to determine the antidermatophyte activity of the extract, formulate a cream with the extract and determine the stability of the cream by measuring the pH, Free-thaw test, Centrifugation and sensitivity to light. We also subjected the formulated cream to temperature variation test at -10, 4, 30, 37 and 45o C.  The percentage yield was 8.5%. The extract contain alkaloid (4.24±0.24%), saponin(1.35±0.39%), tannin (0.45±0.29%) and flavonoid (2.42±0.32%) respectively. The antifungal test results showed the activity against  the dermatophytes to be in the increasing order M.furfur (12± 0.2 mm) < Microsporum audounii (12±0.4 mm) < Trichophyton mentagrophtes (14±0.2mm) < Epidermophyton floccosum     (16±0.2 mm).Temperature stability and Centrifuge testing indicated that the formulations were stable. Light testing indicated no change in the colour of the cream. Our study showed that the plant has high potential as an anti-dermatophyte when formulated as a cream for topical use. The stability of the cream formulations indicated that it can be used for the management of dermatophytosis.

Open Access Original Research Article

Acute and Subchronic Toxicity Studies of Combretum collinum Methanol Root Extract in Albino Rats

S. W. Hassan, A. N. Ukwuani-Kwaja, U. D. Nuhu, R. D. Jabaka

International Journal of Biochemistry Research & Review, Page 9-28
DOI: 10.9734/ijbcrr/2020/v29i1030235

Combretum collinum root extract has been recognized long ago as traditional medicinal plant in curing several diseases among the indigenous people of Alela-land (Zuru), Kebbi State, Nigeria. This research work was carried out to evaluate the toxicological effects of Combretum collinum methanol root extract (CCME) in albino rats. Acute toxicity was performed by a fixed single oral administration at a dose of 10, 100, 1000 mg/Kg and 1600, 2900, 5000 mg/Kg. Subchronic toxicity studies of CCME was conducted at doses of 32, 63, 126 and 253 mg/Kg for 28 days. The result showed that acute administration of CCME resulted at mortality and general behavioral changes at 1000 to 5000 mg/Kg. Therefore, the estimated lethal dose (LD50) of CCME was 316.23 mg/Kg. Subchronic oral administration of CCME revealed a significant (P<0.01) decrease in body weight in rats receiving 63 to 253 mg/Kg throughout the study period compared with the control group. The results also showed a significant (P<0.01) increase in serum ALT, AST, creatinine, potassium and bicarbonate in rats administered with 126 and 253 mg/Kg of the extract. Haematological analysis of the same extract revealed a significant (P<0.01) increase in WBC, HCT, MCV, MCH, MCHC, PLT, LYM and NEUT in rats receiving 126 and 253 mg/Kg only. Histopathological examination of liver revealed severe periportal inflammation, hypertrophy, areas of hydropic changes, cancerous tumor, areas of infiltration and necrosis of the hepatic cells while the kidney showed a mild mesengial hyperplasia, compressed blood vessels, glomerular degeneration, tubular degeneration and tubular widened lumen in rats treated with 63 to 253 mg/Kg. Therefore, caution should be applied as C. collinum root extract has a low mean lethal dose and would be toxic at higher concentrations.

Open Access Original Research Article

RNA Dependent RNA Polymerases of Severe Acute Respiratory Syndrome-Related Coronaviruses- An Insight into their Active Sites and Mechanism of Action

Peramachi Palanivelu

International Journal of Biochemistry Research & Review, Page 29-52
DOI: 10.9734/ijbcrr/2020/v29i1030236

Aim: To analyze the RNA-dependent RNA polymerases (RdRps) of Severe Acute Respiratory Syndrome (SARS)-related coronaviruses (CoVs) to find out the conserved motifs, metal binding sites and catalytic amino acids and propose a plausible mechanism of action for these enzymes, using SARS-CoV-2 RdRp as a model enzyme.

Study Design: Bioinformatics, Biochemical, Site-directed mutagenesis (SDM), X-ray crystallographic and cryo-Electron microscopic (cryo-EM) data were analyzed.

Methodology: Bioinformatics, Biochemical, Site-directed mutagenesis, X-ray crystallographic and cryo-EM data of these enzymes from RNA viral pathogens were analyzed. The advanced version of Clustal Omega was used for protein sequence analysis of the RdRps.

Results: Multiple sequence alignment (MSA) of RdRps from different SARS-related CoVs show a large number of highly conserved motifs among them. Though the RdRp from the Middle Eastern Respiratory syndrome (MERS)-CoV differed in many conserved regions yet the active site regions are completely conserved. Possible catalytic regions consist of an absolutely conserved amino acid K, as in single subunit (SSU) RNA polymerases and most of the DNA dependent DNA polymerases (DdDps). The invariant ‘gatekeeper/DNA template binding’ YG pair that was reported in all SSU DNA dependent RNA polymerases (DdRps), prokaryotic multi-subunit (MSU) DdRps and DdDps is also highly conserved in the RdRps of SARS-CoVs. The universal metal binding motif –GDD- and an additional motif–SDD- are also found in all SARS-CoV RdRps. In stark contrast, the (–) strand RNA viral pathogens like Ebola, rabies, etc. use –GDN- rather than –GDD- for catalytic metal binding. An invariant YA pair (instead of an YG pair) is found in the primases of the SARS-CoVs. The SARS-CoVs RdRps and primases exhibit very similar active site and catalytic regions with almost same distance conservations between the template binding YG/YA pair and the catalytic K. In SARS-CoV RdRps an invariant R is placed at -5 which is shown to play a role in nucleoside triphosphate (NTP) selection and is in close agreement with SSU DdRps (viral family) and DdDps. In primases no such invariant R/K/H is found very close to the catalytic K in the downstream region, as found in RdRp and Nidovirus RdRp-Associated Nucleotidyltransferase (NiRAN) domains. An invariant YA pair is placed in the NiRAN domain instead of an YG pair, and an invariant H is placed at -5 position. Moreover, the Zn binding motif with the completely conserved Cs and a few DxD/DxxD type metal binding motifs are found in the RdRps and NiRAN domain. However, the primases contained only the DXD type metal binding motifs.

Conclusions: The SARS and SARS-related CoV RdRps are very similar as large peptide regions are highly conserved among them. The closer identity between the RdRps of palm civet-CoV and SARS-CoV (CoV-1) suggest their possible link as found between their spike proteins also. The invariant YG and KL pairs may play a role in template binding and catalysis in SARS-CoV RdRps as reported in DdDps. An additional invariant –YAN- motif found in SARS-CoV RdRps may play a crucial role in nucleotide discrimination.

Open Access Original Research Article

α-Klotho as A Novel Biomarkers in Chronic Diabetic Nephropathy

Mohy Eldin Abd EL-Fattah, Taghrid B. El-abaseri, Hegazy Mohamed Abd Elaziz Mohamed

International Journal of Biochemistry Research & Review, Page 53-62
DOI: 10.9734/ijbcrr/2020/v29i1030237

Background: Type 2 diabetes mellitus (DM) is the most common cause of end- stage renal disease. Albuminuria is the foremost commonly utilized marker to anticipate onset of diabetic nephropathy (DN) without sufficient affectability and specificity to identify early DN.

Aim: This study aimed to evaluate plasma α-Klotho as a new biomarker for chronic diabetic nephropathy.

Methods: This cross sectional study included 125 Egyptian subjects attending the out Patients Clinic of the Department of Internal Medicine, 10Th of Ramadan city Health Insurance Hospital and divided into:-control group, patient with diabetic mellitus, patients with Diabetic nephropathy and patient with diabetic nephropathy and other complications. Patients were subjected to measurement of plasma α- Klotho, FBS, HbAIC, serum creatinine, serum urea, serum uric acid, k, Na, serum phosphorus, Albumin: Creatinine Ratio, GFR, Chol, TG, LDL HDL, AST, ALT, T.BIL, D.BIL ALB, TP, GLB and A/G ratio.

Results: Results showed that plasma a-klotho was significantly correlated with hemoglobin A1C, potassium, GFR, Albumin, TP and GLB. Meanwhile, plasma a- klotho was negatively correlated with duration of DM, CR, Urea, UR.A, Na, phosphorus, ACR, Chol, TG, LDL, AST, ALT, T.BIL, and D.BIL. However, there were no significant correlations between plasma α -klotho and FBS, HDL and A/G

ratio. At cut-off level ≥2.6, plasma α -klotho had 95% sensitivity and 81% specificity for diagnosing diabetic nephropathy.

Conclusion: α-klotho may be the chronic diabetic nephropathy markers for predicting renal injury in patients with type 2 diabetes.

Open Access Original Research Article

Plasma Cyclophilin A as a Novel Biomarker in Chronic Nephropathy

Mohy Eldin Abd EL-Fattah, Taghrid B. El-abaseri, Hegazy Mohamed Abd Elaziz Mohamed

International Journal of Biochemistry Research & Review, Page 63-72
DOI: 10.9734/ijbcrr/2020/v29i1030238

Background: Type 2 diabetes mellitus (DM) is the most common cause of end- stage renal disease. Albuminuria is the foremost commonly utilized marker to anticipate onset of diabetic nephropathy (DN) without sufficient affectability and specificity to identify early DN.

Aim: This study aimed to evaluate Plasma cyclophilin A (CypA) as a new biomarker for early DN.

Methods: This cross sectional study included 125 Egyptian subjects attending the out Patients Clinic of the Department of Internal Medicine, 10Th of Ramadan city Health Insurance Hospital and divided into-:control group, patient with diabetic mellitus, patients with Diabetic nephropathy and patient with diabetic nephropathy and other complications. Patients were subjected to measurement of plasma cyclophyline A, FBS, HbAIC, serum creatinine, serum urea, serum uric acid, k, Na, serum phosphorus, Albumin:Creatinine Ratio, GFR, Chol, TG, LDL HDL, AST, ALT, T.BIL, D.BIL ALB, TP, GLB and A/G ratio.

Results: Results showed that Cyclophilin A was significantly correlated with duration of DM, CR, Urea, UR.A, Na, phosphorus, ACR, Chol, TG, LDL, AST, ALT, T.BIL, D.BIL. Meanwhile, Cyclophilin A was negatively correlated with HA1C, K, GFR, HDL, ALB, TP, GLB and A/G ratio. At cut-off level ≥84.14, cyclophilin A had 91% sensitivity and 62% specificity for diagnosing diabetic nephropathy.

Conclusion: CypA can be used as an early marker for DN as we found early significant high levels of urinary CypA in diabetic patients with stage 2 DN even before the appearance of albuminuria.