Cytotoxic, Antibacterial, and Leishmanicidal Activities of Paullinia pinnata (Linn.) Leaves

Background: Paullinia pinnata leaves are employed traditionally for the treatment of various ailments which are of biological origin. Objectives: The aim of this study was to explore cytotoxic, antibacterial, and antileishmanial properties of the leaves of Paullinia pinnata using in vitro models. Methods: Brine shrimp lethality bioassay was used to determine the cytotoxic activity of the methanol leaf extract of Paullinia pinnata. The activity of the extract against the growth of cultured Leishmania major (DESTO) promastigotes was used to investigate the leishmanicidal activities. The agar well diffusion method was used to investigate the antibacterial activity against Salmonella typhi, Pseudomonas aeruginosa, Shigella flexneri, Escherichia coli, Staphylococcus aureus, and Bacillus subtilis. Results: The methanol leaf extract of P. pinnata had no activity against Artemia salina (brine shrimp) and L. major. It showed a non-significant activity against E. coli and B. subtilis and no activity against P. aeruginosa, S. flexneri, S. typhi, and S. aureus. Conclusion: The methanol leaf extract of P. pinnata did not exhibit any cytotoxic and anti-leishmanial properties. Moreover, the activity against various species of bacteria was not significant.

The Effect of Venom of Egyptian Spitting Cobra Naja nubiae on Vascular Permeability of Hepatic and Renal Tissues

Background: Among venomous elapid snakes, cobras have the highest public awareness, as their venom represents a combination of proteins, peptides, and enzymes that have a range of biochemical and pharmacological roles and are also the main constitutes of biological activity and lethal toxicity. Objectives: The study aimed to evaluate the effect of the venom of Egyptian Spitting Cobra, Naja nubiae, on the vascular permeability based on the extravasation of the azo dye Evans blue (EB) into the tissues of the liver and kidneys of animals envenomed with low (¼ LD50; 0.32 mg/kg) and high (½ LD50; 0.65 mg/ kg) doses at three sampling times (30, 120, 360 min) post-injection of the venom. Methods: Fifty-four adult male Albino rats (8 weeks old and 180±2 0 g body weight) were divided into three main groups (n=6). In the control group, rats were subcutaneously (SC) injected with saline solution. Envenomed groups were SC injected, one group with 0.32 mg/kg and the other group with 0.65 mg/kg body weight of crude venom, respectively. Rats were I.V injected with EB dye 20 minutes before SC injection with saline solution as control animals and with Naja nubiae venom as treatment groups. Results: The results illustrated a high significant rate of EB extravasation to hepatic and renal tissues by the colorimetric determination of EB dye concentration. Conclusion: The venom of Naja nubiae can cause increased hepatic and renal vascular permeability which may explain the inflammatory effect induced by this venom.

Audit of Reporting Errors in Biochemistry Section of NABL Accredited Laboratory in a Tertiary Care Teaching Hospital

Background: In the modern era of tremendous automation in analytical processes, reporting errors have been reduced significantly. Therefore, the focus has been shifted to identifying the extra analytical causes of errors in the laboratory. Objectives: This study aimed to audit major clinical decisions affecting quality indicators (i.e., reporting errors and error prevention) by adhering to ISO 15189 (2012) and National Accreditation Board for Testing and Calibration Laboratories (NABL) (112) requirements. Methods: The records of the reporting errors were maintained from the biochemistry section of the central clinical laboratory (CCL) and analyzed based on the aim of this study. Then, the root cause analysis was performed, and the data was collected and audited from November 2015 to July 2020. Results: The total number of reporting errors between the mentioned periods were 132, with an incidence of 1 error per 384 processed samples on the day of observing the reporting error. In general, 22 (16.67%), 16 (12.12%), and 94 (71.21%) cases were pre-analytical, analytical, and post-analytical errors, respectively. The incidence of the post-analytical error was noted to be more since they were all typographical errors. Conclusion: Overall, transcriptional or typographical errors were found to be the main causes of reporting errors. In our clinical laboratory, we are attempting to minimize these errors by pre-validating the results by senior technicians and faculty prior to the typing and approval. These avoidable errors can be minimized by the continuous training of laboratory staff. Up-gradation to automated data collection information management systems are of great hope for preventing such errors.

Blood Pressure Regulating and Antioxidant Potentials of Theobroma Cacao Pod Husk Protein Hydrolysates

Background: Agrowastes like Theobroma cacao (Cocoa) pod husk can be used to prepare bioactive peptides with various bio-functionalities. Objectives: This study aimed to investigate antioxidant and angiotensin converting enzyme I (ACE) inhibitory peptides contained in Theobroma cacao (cocoa) pod husks – an agro-waste. Methods: Protein isolated from cocoa pod husk was enzymatically digested with alcalase, pepsin, and trypsin. ACE inhibition, kinetics of ACE inhibition, and antioxidant properties of the cocoa pod husks hydrolysates were evaluated in vitro. Results: Trypsin and alcalase hydrolysates displayed higher peptide yields (63.1% and 61.2%) than pepsin hydrolysate (61.2%). However, no significant difference (P>0.05) was observed in the degree of hydrolysis (DH) of the three proteases on cocoa pod husk protein. Methionine, lysine, and cysteine were the amino acid residues presented in cocoa pod husk hydrolysates. A concentration-dependent ACE inhibition by cocoa pod husk hydrolysates was observed. The highest ACE inhibitions of 84.4%, 81.5%, and 73.5% were obtained at 2.0 mg/mL of pepsin, trypsin, and alcalase hydrolysates, respectively, with the minimum IC50 value of 0.36 mg/mL obtained for trypsin hydrolysate. An uncompetitive and mixed-type inhibition was obtained from double reciprocal plots of alcalase and pepsin as well as trypsin cocoa pod husk protein hydrolysates. The Ki values of ACE inhibition for pepsin, trypsin, and alcalase hydrolysates were 3.05, 2.19, and 3.57 mg/mL, respectively. A concentration-dependent increase in the scavenging of 2,2-diphenyl-1-picrylhydrazyl and superoxide radicals as well as ferric reducing antioxidant power were recorded for the cocoa pod husk hydrolysates. Conclusion: Trypsin and alcalase cocoa pod husk protein hydrolysates could be an effective source of a natural ACE inhibitor and antioxidant.

Molecular Analysis of β-Globin Mutations Among β-Thalassemia Patients in Hamadan

Background: β-Thalassemia (βT) is one of the most common genetic diseases. The specific mutation profile of that region can be identified by determining the specific mutations of each region and ethnicity. Objectives: This study investigated the β-globin mutations in patients with βT in Hamadan. Methods: This cross-sectional study was performed on 47 βT carriers. In the present study, the polymerase chain reaction (PCR)-sequencing technique was used to confirm βT carriers, and data were analyzed with SPSS-16 at a 95% confidence level. Results: In general, 164 individuals (81 men and 83 women) suspected of having thalassemia were examined, where 28.7 % (n=47) of them were identified by PCR-sequencing with βT carriers (48.8% male and 53.2% females). Hemoglobin beta (HBβ): c.251 del, HBβ: c.27dupG, and HBβ: c.92+5G>A mutations had the greatest effect on mean corpuscular volume (MCV) reduction, mean corpuscular HB (MCH) reduction, and HbA2 increment, respectively. The most common mutation in both males and females was the same (HBβ: c.315+1G>A). Conclusion: According to the results, the most common mutations in the diagnosis of βT in Hamadan were serially HBβ: c.315+1G>A mutation and HBβ: c.25-26del, HBβ: c.112del, HBβ: c.20A>T, HBβ: 92+6T>C, and HBβ: c.316-106C>G.

Unexpected Detection of Abnormal Hemoglobin Variants During Routine HbA1c Analysis by HPLC Technique: Challenges and Opportunities in HbA1c Testing

Background: Glycated Hemoglobin A1c (HbA1c) assay is most widely used in diabetic patients for assessing long-term control of glycemia. The presence of hemoglobin variants may be an incidental finding and can interfere with HbA1c measurements. The aim of the present study is to investigate the prevalence and impact of interference of different abnormal hemoglobin variants on HbA1c measurements during routine HbA1c testing. Methods: A total of 12,092 HbA1c samples were collected from January to August 2018. HbA1c quantification was carried out on a Variant II Bio-Rad’s HPLC analyzer. Abnormal chromatograms were further analyzed using the extended-run high-pressure liquid chromatography (HPLC) analysis in the A2/F mode. Results: The samples were examined for presence of abnormal variants. Samples producing abnormal chromatograms were further analyzed in A2/F mode to characterize hemoglobin variants. Abnormal variants were identified in 126 (1%) samples, and 74 (0.59%) sickle cell traits (SCT) were the most common variant in our findings. Moreover, 30 (0.24%) cases were eluted in the variant window in A1c mode, which on further analysis were found to be Hb E & Hb D traits. Furthermore, 3 (0.02%) cases were eluted at a RT <1 min as (unknown) and identified as Hb H. Also,19 (0.15%) samples were eluted in the P3 window at different retention times. Conclusion: Observing each chromatograph after the analysis can help us in identifying silent hemoglobin variants in routine HbA1c testing. Knowledge and awareness of common hemoglobin variants affecting measurement of HbA1c is imperative to avoid reporting of falsely low HbA1c values in diabetic population.

Glycemic Index and Glycemic Load of Four Local Fruits of Benin Among Apparently Healthy Young Adult Subjects

Background: Daily consumption of fruits is recommended due to their positive impact on the control of glycemia, cholesterol and coronary heart disease. Objectives: This study aimed to determine the glycemic index and glycemic load (GL) of four local fruits grown in Benin, namely papaya, pineapple, watermelon and grafted mango, among apparently healthy young adult subjects. Methods: This research work, being an interventional study of quasi-experimental category, involved 33 voluntary adult subjects (mean age: 23.4±1.9 years; mean body mass index: 21.38±1.89 kg/m2 ) distributed into 4 groups. The subjects of each group consumed the reference food (25 g of glucose or 50 g of white bread) twice a week with an interval of one week, and then a serving equivalent to 25 g of carbohydrates of each tested fruit in the morning after a 12-hour fasting on the evening. Plasma glucose was measured at 0, 15, 30, 45, 60, 90, and 120 minutes after food ingestion. Data were analyzed by one-way analysis of variance (ANOVA, SPSS, 26). The P < 0.05 was regarded as the significance level. Results: The incremental area under the curve mean value in mmol.L-1.min-1 of pineapple (89.21±21.75) was higher (P <0.001) than those of mango (34.71±13.62), papaya (23.46±15.06) and watermelon (20.30±16.47). The mean glycemic index of mango (117.09±58.32) was significantly higher (P =0.007) than the ones of pineapple (52.97±29.87), papaya (46.77±45.77), and watermelon (41.04±34.06). The mean GL of mango (16.28±8.11) was significantly more elevated (P =0.001) than the ones of papaya (3.41±3.34), pineapple (6.36±3.58), and watermelon (2.54±2.11). Conclusion: Watermelon, papaya and pineapple may therefore be recommended for safe consumption in accordance with dietary guidelines.

Effect of Ethanol Extract of Aframomum angustifolium Seed on the Induction of Nephrotoxicity by Bromate in Wistar Rats

Background: The continued use of bromate due to its oxidizing property poses health hazards since it is an established nephrotoxic agent. Objectives: This study evaluated the capacity of the ethanol extract of Aframomum angustifolium seeds to ameliorate the nephrotoxicity of potassium bromate in Wistar rats. Methods: In stage I of this study, the main phytochemical groups in the seeds were quantified using spectrophotometric procedures. The acute and sub-chronic toxicities of the extract were studied by monitoring physical and biochemical parameters in stage II. In stage III, the reno-protective effect of the extract were determined by administering 350 and 750 mg/kg bw of the extract with 30 mg/kg bw potassium bromate orally. The reno-protective study lasted for 56 days and the effect of treatment on biomarkers was determined on days 28 and 56. Results: The phytochemical groups (i.e., alkaloids, flavonoids, saponins, tannins, ascorbic acid, and alpha-tocopherol) were detected in the seeds. The acute and sub-chronic oral administration of the extract did not induce any significant toxic reactions across the studied concentrations. The sub-chronic administration of the extract reduced average weight gain in the treated groups. The obtained results in the reno-protective and histological studies indicated that the seed extract offers protection against the induced oxidative assault by bromate. Conclusion: In general, the co-administration of the ethanol extract of A. angustifolium seeds with bromate can reduce its nephrotoxicity in a dose-dependent manner.

Assessment of Antioxidant and Antimicrobial Activities of Silver Nanoparticles Biosynthesized by Haplophyllum Obtusifolium

Background: Plants comprise great antioxidant sources as a result of their redox and biochemical components, which are rich in secondary metabolites such as phenolic acids, flavonoids, and other constituents. Haplophyllum obtusifolium from polygonaceae is widely used for preventing and managing diabetes. This study investigated the antibacterial and antioxidant activities of silver nanoparticles (AgNPs) biosynthesized by H. obtusifolium. Methods: The aerial parts of H. obtusifolium were gathered from the north of Khorasan Razavi province, Iran and desiccated at the chamber temperature. The shoots were powdered by grinding, 5 g of the powder was mixed with 250 mL of deionized water, and the resultant blend was then filtered. Bactericidal properties and antioxidant activity of the nanoparticles were assessed using disk diffusion and DPPH (2, 2-diphenyl-1-picrylhydrazyl) tests, respectively. Results: The results of this study showed that the biosynthesized nanoparticles exhibited antibacterial activity against a gram-negative (Klebsiella pneumoniae) bacterium, but they had no effects on gram-positive Staphylococcus epidermidis. Antioxidant test results showed that these nanoparticles were capable of eliminating DPPH radicals in a concentration-dependent manner so that a more potent antioxidant activity was seen in higher concentrations of the nanoparticles. Conclusion: Our results suggested that H. obtusifolium can be used as a key source of antioxidants/ antimicrobial agents in food and pharmaceutical industries.

Assessment of the Anti-apoptotic Effects of Peganum harmala Leaf Extract on Type 2 Diabetes in the Kidney of Male Wistar Rats

Background: Growing evidence has shown that the apoptosis of cells plays an important role in the advancement of the Diabetic nephropathy (DN). Objectives: This study attempted to discover the therapeutic potential of Peganum harmala leaf extract in the apoptosis of diabetic kidney disease. Methods: In the present experimental research, 32 male Wistar rats were studied, and diabetes was induced by streptozotocin (STZ) (65 mg/kg). The animals were randomly divided into four groups (n=8, in each group) as follows: control, diabetic, control+leaf extract, diabetic+leaf extract. For our purposes, the methanolic extract of P. harmala leaves (150 mg/kg) was given by gavage for 28 days. Flow cytometry and real-time polymerase chain reaction (PCR) analyses were utilized to determine the percentages of apoptotic cells. Also, histological alterations and blood biochemical parameters were evaluated. Results: The P. harmala leaf extract has a high amount of flavonoids (25.84%), a lower percentage of alkaloids (0.14%), and some antioxidant properties. Serum urea (P<0.001) and apoptosis (P<0.05) significantly elevated in diabetic rats relative to the control ones. The mean of fasting blood creatinine, urea, and albumin level was not significantly changed in diabetic+leaf extract rats as compared to the diabetic ones. Histopathological results also displayed that diabetic complications in the kidney could not be improved following treatment by the leaf extract of P. harmala. In addition, the leaf extract could not significantly reduce the apoptosis and caspase-3 expression compared to diabetics in renal cells. Conclusion: Based on our findings, the leaf extract of P. harmala is unable to inhibit apoptosis in the diabetic kidney model.