Immuno-Kachiks formula immunomodulates and ameliorates hepatic damage induced by monosodium glutamate in rats.

The immune system plays a vital role in controlling liver fibrosis and enhancing the pathogenesis of liver inflammation. Monosodium glutamate is a common flavor-enhancement food additive. This study evaluated the immunomodulatory and hepato-curative effects of the Immuno-Kachiks polyherbal formulation against monosodium glutamate-induced immune suppression and hepatic damage in rats. Monosodium glutamate was given orally at a 2000 mg/kg dose to male Wistar rats for three months to induce liver damage and immune suppression. After three months of successful induction, three groups were separately administered orally with Immuno-Kachiks formula at 400, 800, and 1500 mg/kg/day for 28 days. At the end of the treatment period, liver and blood samples were collected for histological and biochemical analysis. The lymphocyte count remained significantly low while the neutrophil count and the neutrophil-to-lymphocyte ratio increased significantly, despite the cessation of monosodium glutamate ingestion for 28 days. The Immuno-Kachiks formula (IKF) significantly increased the lymphocyte count, reduced the neutrophil-to-lymphocyte ratio, and normalized the neutrophil count. Neither monosodium glutamate nor the IKF significantly caused alpha-fetoprotein levels to rise or fall below normal. High doses (800 and 1500 mg/kg) of the Immuno-Kachiks formula significantly raised serum levels of aspartate aminotransferase, alkaline phosphatase, and total bilirubin. 1500 mg/kg of the IKF caused mild liver inflammation. The IKF restored the liver morphologic alterations observed in monosodium glutamate-induced liver damage in rats. The results suggest that the Immuno-Kachiks herbal formulation is a potential curative agent for early-stage liver damage and could restore suppressed adaptive immunity.

Antimicrobial resistance and rational prescription practices: knowledge, perceptions and confidence of health profession interns in Uganda.

Background: Antimicrobial resistance (AMR) is significantly driven by misuse and overuse of antibiotics. Graduate health profession interns often prescribe antimicrobials under minimum supervision. Objectives: This study explored the knowledge, perceptions and confidence of health profession interns in Uganda regarding AMR and rational prescription practices. Methods: This was a cross-sectional survey employing quantitative techniques carried out between October and November 2022 at six tertiary hospitals in Uganda. Health profession interns including doctors, nurses, midwives and pharmacists were recruited as study participants. Data were collected using online Kobo toolbox software. Data analysis was performed using STATA (StataCorp) version 16. Bivariate analysis and multivariable logistic regression were performed. P < 0.05 was considered statistically significant. Results: We recruited 281 participants with a mean age of 27 ±â€Š3.8 years, of which few (n = 53; 19%) had good knowledge about AMR and rational prescription. The use of professional organization guidelines as a source of information was significantly associated with good knowledge (adjusted OR = 1.9; 95% CI: 1.0-3.5; P = 0.046). Nurses had the least knowledge compared with doctors and pharmacists. Continuous medical education (99%) and availability of clinical guidelines (98%) were identified as the most helpful intervention to improve knowledge. Most participants were confident about accurately diagnosing infections and sepsis and selecting appropriate antimicrobials. Conclusions: Continuous medical education and availability of clinical and professional organization guidelines should be leveraged to improve the knowledge of AMR and rational prescription among health profession interns. Their high confidence in rational prescription practices should be pivotal to the fight against AMR.

A review of efficacy and safety of Ugandan anti-malarial plants with application of RITAM score.

BACKGROUND: Malaria, a treatable disease mainly caused by Plasmodium falciparum has remained a health challenge in Africa, a continent that accounted for 96% of total global cases and deaths in 2021. Uganda, a malaria endemic country is experiencing malaria parasite resistance to some of the drugs used in the artemisinin-based combination therapy (ACT). In an effort to prioritize herbal medicines for new product development, this review synthesized the available safety and efficacy literature on the Ugandan anti-malarial plants to suggest most effective herbal plants. METHODS: Literature was exhaustively searched using engines and databases, such as Google scholar, Pubmed, and Scopus-indexed journals during the period of June 2020-December 2021. In the first phase, information on ethnobotanical uses of anti-malarial plants in Uganda was gathered and synthetized to generate a list of plants, followed by data on anti-malarial efficacy (both in vitro and in vivo) on each listed plant. Minimum inhibitory concentrations (µg/ml), and % parasite suppression for every plant were scored using The Research Initiative on Traditional and Antimalarial Methods (RITAM) scoring system. The best twenty (20) plants were evaluated for acute safety (LD50) data in rat model, plant parts used, ease of cultivation, presence of clinical studies and other relevant factors for suggesting the best three (3) plants for future anti-malarial product development. RESULTS: Over one hundred twenty-six (126) plant species are used in Uganda for treatment of malaria in local communities. Out of these, about 33% (41) have been studied for efficacy and safety, with Artemisia annua and Vernonia amygdalina being the most extensively studied and among the best twenty (20) anti-malarial plants in Uganda. Both are limited by parasite recrudescence in clinical studies. Microglossa pyrifolia, a very potent plant (IC50 = 0.03 - 0.05 µg/ml has potential to penetrate the liver and could ameliorate the challenge of recrudescence if combined with A. annua and V. amygdalina in a polyherbal formulation. CONCLUSION: There are many plants with promising potential for malaria treatment in Uganda and a herbal combination of A. annua, V. amydalina and M. pyrifolia could offer the next herbal ACT if carefully studied and developed.

Hypoglycemic, Antihyperglycemic, and Toxic Effects of Physalis peruviana L. Aqueous and Methanolic Leaf Extracts in Wistar Rats.

Background: Physalis peruviana L. (Solanaceae) is a plant widely used in traditional medicine systems to manage various diseases, including diabetes mellitus, which remains a global health problem in developing and developed countries. This study aimed to scientifically evaluate its antidiabetic bioactivity and short-term toxicity in rats. Methods: We prepared various doses (100, 200, 400 mg/kg) of aqueous and methanolic leaf extracts for the antidiabetic study, and a dose of 2000 mg/Kg was prepared for the acute toxicity test. The first group that evaluated the hypoglycemic effect consisted of forty normoglycemic Wistar rats aged 7-8 months old with a weighted average of 265.8 ± 24.6 g. The second group consisted of intraperitoneal glucose-loaded male animals to evaluate the antihyperglycemic effect. The third group contained two groups of normoglycemic female rats (n = 3), aged 3 and 4 months old (weight average: 187.45 ± 14.82 g), treated for 14 days with aqueous and methanolic extracts (2 g/kg b.w) to assess mortality and toxic effects. Blood samples were taken at 30, 60, 90, and 120 min post-treatment in hypoglycemic and antihyperglycemic evaluations. Glibenclamide (5 mg/kg) was used as a reference drug. The control animals in each group did not receive the extracts. Results: In hypoglycemic rats, 100 mg/kg of aqueous and methanolic extracts significantly lowered the fasting blood glucose level by 13.92% (p < 0.0001) and 21.95% (p < 0.01), respectively, compared to the control group. In glucose tolerance test group, methanolic extracts significantly reduced hyperglycemia by 54.55% (p < 0.0001), 46.50% (p < 0.0001), 39.78% (p < 0.0001) at 400, 200 and 100 mg/kg b.w, respectively, compared to control; aqueous extract 400 mg/kg reduced hyperglycemia by 39.44% (p < 0.05). At the 2000 mg/kg dose, leaf aqueous and methanolic extracts did not show any signs of intoxication and mortality. Conclusion: Crude aqueous and methanolic leaf extracts of P. peruviana ambrosioides appeared safe at 2000 mg/kg and have bioactivity in controlling the blood glucose levels, supporting their use in treating diabetes.

Effects of the Oral Administration of Aqueous and Methanolic Leaf Extracts of Chenopodium ambrosioides L. (Amaranthaceae) on Blood Glucose Levels in Wistar Rats.

Background: Diabetes mellitus is a metabolic disorder that poses a major global health threat. The current diabetes mellitus uses insulin and oral hypoglycemic agents, which have limitations, including adverse effects and secondary failures. Herbal medicine is being evaluated for its role in the pharmacotherapy of diabetes. This study was aimed to assess the anti-diabetic potential and short-term toxicity level of Chenopodium ambrosioides collected from Bukavu in Democratic Republic of Congo. Methods: Leaves of C. ambrosioides were extracted by infusion and maceration with distilled water and 95% methanol, respectively. Hypoglycemic and antihyperglycemic potentials of the aqueous and methanolic were investigated in normoglycemic and intraperitoneal glucose-loaded rats at 100, 200, and 400 mg/kg body weight. An oral acute toxicity test was carried out on healthy female Wistar rats. Results: Acute toxicity test showed the mean lethal dose (LD50) for both aqueous and methanol extracts of C. ambrosioides to be more than 2000 mg/kg. The group treated with glibenclamide (5 mg/kg b.w) and aqueous extract of the plant (200 mg/kg b.w) showed a significant reduction (p< 0.0001 and p< 0.05) of fasting blood glucose by 46.91% and 16.72%, respectively, compared to control and all other treatment groups. In acute conditions, a single oral administration of the aqueous and methanolic extracts lowered fasting blood glucose in rats. Any manifestation and signs of toxicity and mortality have been recorded for 14 days of observation. Conclusion: Leaf aqueous and methanolic extracts of C. ambrosioides appeared safe at 2000 mg/kg. The plant demonstrated some anti-diabetic potential in rats, explaining its use as an anti-diabetic remedy locally.

Prevalence and Factors Associated with Non-Adherence to Antidiabetic Medication Among Patients at Mbarara Regional Referral Hospital, Mbarara, Uganda.

BACKGROUND: Non-adherence is a major concern in the treatment of diabetes mellitus and undermines the goals of treatment. The objective of this study was to determine the magnitude of non-adherence and its contributing factors among diabetes mellitus patients attending the diabetes mellitus clinic at Mbarara Regional Referral Hospital. OBJECTIVE: To assess prevalence and factors contributing to non-adherence to antidiabetic medication among diabetes mellitus patients in the diabetic clinic at Mbarara Regional Referral Hospital. METHODS: A descriptive cross-sectional study was adopted at the diabetes clinic, Mbarara Regional Referral Hospital, between July and October 2020. Study participants were systemically sampled, and data regarding their medication non-adherence was collected using a structured questionnaire, based on the Hill-Bone medication adherence scale. Data entry was done using Microsoft Excel Version 2010, and analysis was carried out using STATA version 13. The odds ratio was used to determine the strength of association between diabetic medication non-adherence and associated factors. The cutoff value for all statistical significance tests was set at p < 0.05 with a confidence interval of 95%. RESULTS: A total of 257 participants were recruited with a 100% response rate. More than one-third (98, 38.1%) of the participants were non-adherent to their antidiabetic medication. Age above 60 years (AOR = 6.26, 95% CI = 1.009-39.241, P = 0.049) and duration of diabetes mellitus above 5 years (AOR = 1.87, 95% CI = 1.034-3.392, P = 0.038) were independently associated with non-adherence to antidiabetic medication. CONCLUSION: The prevalence of non-adherence to antidiabetic medication was higher than that revealed in previous studies in Uganda. Patients with age above 60 years were six times more likely to be non-adherent to their anti-diabetic medications. Patient education is important to address the challenges of medication non-adherence.

Antidiabetic Medicinal Plants Used in Democratic Republic of Congo: A Critical Review of Ethnopharmacology and Bioactivity Data.

Several studies have been conducted and published on medicinal plants used to manage Diabetes Mellitus worldwide. It is of great interest to review available studies from a country or a region to resort to similarities/discrepancies and data quality. Here, we examined data related to ethnopharmacology and bioactivity of antidiabetic plants used in the Democratic Republic of Congo. Data were extracted from Google Scholar, Medline/PubMed, Scopus, ScienceDirect, the Wiley Online Library, Web of Science, and other documents focusing on ethnopharmacology, pharmacology, and phytochemistry antidiabetic plants used in the Democratic Republic of Congo from 2005 to September 2021. The Kew Botanic Royal Garden and Plants of the World Online web databases were consulted to verify the taxonomic information. CAMARADES checklist was used to assess the quality of animal studies and Jadad scores for clinical trials. In total, 213 plant species belonging to 72 botanical families were reported. Only one plant, Droogmansia munamensis, is typically native to the DRC flora; 117 species are growing in the DRC and neighboring countries; 31 species are either introduced from other regions, and 64 are not specified. Alongside the treatment of Diabetes, about 78.13% of plants have multiple therapeutic uses, depending on the study sites. Experimental studies explored the antidiabetic activity of 133 plants, mainly in mice, rats, guinea pigs, and rabbits. Several chemical classes of antidiabetic compounds isolated from 67 plant species have been documented. Rare phase II clinical trials have been conducted. Critical issues included poor quality methodological protocols, author name incorrectly written (16.16%) or absent (14.25%) or confused with a synonym (4.69%), family name revised (17.26%) or missing (1.10%), voucher number not available 336(92.05%), ecological information not reported (49.59%). Most plant species have been identified and authenticated (89.32%). Hundreds of plants are used to treat Diabetes by traditional healers in DRC. However, most plants are not exclusively native to the local flora and have multiple therapeutic uses. The analysis showed the scarcity or absence of high-quality, in-depth pharmacological studies. There is a need to conduct further studies of locally specific species to fill the gap before their introduction into the national pharmacopeia.

Ethnotherapeutic Uses and Phytochemical Composition of Physalis peruviana L.: An Overview.

BACKGROUND: Plant-derived medicines are widespread and continue to increase in traditional and modern medicine, especially in developing countries. Physalis peruviana L. is among the most used plants in conventional medication worldwide. This review aimed to highlight the ethnotherapeutic uses and phytochemical status of identified compounds in P. peruviana. METHODS: Data were collected from Google Scholar, PubMed/Medline, SciFinder, Science Direct, Scopus, the Wiley Online Library, Web of Science, and any other helpful search engine using Physalis peruviana as the primary keyword. RESULTS: Some countries, worldwide, use P. peruviana in their traditional medicine system to manage diverse ailments, mainly diseases and gastrointestinal tract disorders (25.33%). Leaf was the mostly used part (49.28%), prepared by decoction (31.58%) and overall administrated orally (53.57%) as the main route of admission. Around 502 phytoconstituents were identified in different plant parts, especially fruit (38.19%) ethanol/ethyl acetate extract. In most cases (36.17%), the solvent of the extract was not specified. Several phytochemical classes were found in the plant, especially terpenes (26.09%) and phenolic compounds (14.94%). Esters were also abundant (11.55%). In the terpenes category, carotenoids were the most abundant (11.15% followed by monoterpenes (8.76%) and diterpenes (3.18%). However, flavonoids (5.17%) followed by cinnamic acid derivatives (3.99%), monophenolic compounds (1.79%), and phenolic acids (1.33 M) are the most reported phenolic compounds. Hexadecanoic acid (palmitic acid) was the most cited (five times). CONCLUSION: P. peruviana plays an essential role in managing diseases in some countries and is rich in chemical compounds, which need to be isolated and investigated pharmacologically before clinical trials.

Registration and local production of essential medicines in Uganda.

BACKGROUND: Universal access to high quality essential medicines is critical to sustainable development (SDG 3.8). However low- and middle-income countries struggle to ensure access to all medicines on their national essential medicines lists (EML). Market registration is the first step in determining both access and availability yet the extent to which essential medicines are registered for use at country level is not known. Companies apply for a marketing authorisation, however low price or lack of a market is a disincentive. Local production has been promoted to ensure availability of essential medicines but research in this area is also limited. METHODS: The study took place between 2011 and 2015. We systematically examined the registration status of medicines and vaccines listed in the Ugandan 2012 EML and conducted 20 interviews with regulators, ministry of health representatives, donors, and pharmaceutical producers and analysed quality assurance issues affecting registration, procurement, and local production of medicines in Uganda. In 2017 we conducted a further three interviews to clarify issues around non-registration of essential medicines highlighted by our analysis. RESULTS: Of the 566 essential medicines and vaccines nearly half (49%; 275/566) had no registered product in 2012. Of the 3130 registered products, just over a quarter (28%; 880/3130) were listed on the EML. Six local producers had registered 138 products of which 40 corresponded to 32 unique essential medicines. Interviews highlighted alternative routes to availability other than registration. Local producers faced considerable barriers to achieving international quality standards required for international procurement of medicines for the domestic market. CONCLUSIONS: Monitoring and audit of the registration of essential and non-essential medicines should be a priority nationally and, regionally through harmonisation of registration requirements in the East African Community. National and regional manufacturing plans should consider local production of unregistered essential medicines.

Metabolomic Profiling of the Synergistic Effects of Melittin in Combination with Cisplatin on Ovarian Cancer Cells.

Melittin, the main peptide present in bee venom, has been proposed as having potential for anticancer therapy; the addition of melittin to cisplatin, a first line treatment for ovarian cancer, may increase the therapeutic response in cancer treatment via synergy, resulting in improved tolerability, reduced relapse, and decreased drug resistance. Thus, this study was designed to compare the metabolomic effects of melittin in combination with cisplatin in cisplatin-sensitive (A2780) and resistant (A2780CR) ovarian cancer cells. Liquid chromatography (LC) coupled with mass spectrometry (MS) was applied to identify metabolic changes in A2780 (combination treatment 5 µg/mL melittin + 2 µg/mL cisplatin) and A2780CR (combination treatment 2 µg/mL melittin + 10 µg/mL cisplatin) cells. Principal components analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) multivariate data analysis models were produced using SIMCA-P software. All models displayed good separation between experimental groups and high-quality goodness of fit (R²) and goodness of prediction (Q²), respectively. The combination treatment induced significant changes in both cell lines involving reduction in the levels of metabolites in the tricarboxylic acid (TCA) cycle, oxidative phosphorylation, purine and pyrimidine metabolism, and the arginine/proline pathway. The combination of melittin with cisplatin that targets these pathways had a synergistic effect. The melittin-cisplatin combination had a stronger effect on the A2780 cell line in comparison with the A2780CR cell line. The metabolic effects of melittin and cisplatin in combination were very different from those of each agent alone.

A method for the analysis of sugars in biological systems using reductive amination in combination with hydrophilic interaction chromatography and high resolution mass spectrometry.

Separation of sugar isomers extracted from biological samples is challenging because of their natural occurrence as alpha and beta anomers and, in the case of hexoses, in their pyranose and furanose forms. A reductive amination method was developed for the tagging of sugars with the aim of it becoming part of a metabolomics work flow. The best separation of the common hexoses (glucose, fructose, mannose and galactose) was achieved when 2H5-aniline was used as the tagging reagent in combination with separation on a ZICHILIC column. The method was used to tag a range of sugars including pentoses and uronic acids. The method was simple to perform and was able to improve both the separation of sugars and their response to electrospray ionisation. The method was applied to the profiling of sugars in urine where a number of hexose and pentose isomers could be observed. It was also applied to the quantification of sugars in post-mortem brain samples from three control samples and three samples from individuals who had suffered from bipolar disorder.

Metabolomic Profiling of the Effects of Melittin on Cisplatin Resistant and Cisplatin Sensitive Ovarian Cancer Cells Using Mass Spectrometry and Biolog Microarray Technology.

In the present study, liquid chromatography-mass spectrometry (LC-MS) was employed to characterise the metabolic profiles of two human ovarian cancer cell lines A2780 (cisplatin-sensitive) and A2780CR (cisplatin-resistant) in response to their exposure to melittin, a cytotoxic peptide from bee venom. In addition, the metabolomics data were supported by application of Biolog microarray technology to examine the utilisation of carbon sources by the two cell lines. Data extraction with MZmine 2.14 and database searching were applied to provide metabolite lists. Principal component analysis (PCA) gave clear separation between the cisplatin-sensitive and resistant strains and their respective controls. The cisplatin-resistant cells were slightly more sensitive to melittin than the sensitive cells with IC50 values of 4.5 and 6.8 µg/mL respectively, although the latter cell line exhibited the greatest metabolic perturbation upon treatment. The changes induced by melittin in the cisplatin-sensitive cells led mostly to reduced levels of amino acids in the proline/glutamine/arginine pathway, as well as to decreased levels of carnitines, polyamines, adenosine triphosphate (ATP) and nicotinamide adenine dinucleotide (NAD+). The effects on energy metabolism were supported by the data from the Biolog assays. The lipid compositions of the two cell lines were quite different with the A2780 cells having higher levels of several ether lipids than the A2780CR cells. Melittin also had some effect on the lipid composition of the cells. Overall, this study suggests that melittin might have some potential as an adjuvant therapy in cancer treatment.

Effect of Bee Venom and Its Fractions on the Release of Pro-Inflammatory Cytokines in PMA-Differentiated U937 Cells Co-Stimulated with LPS.

The venom of Apis mellifera (honey bee) has been reported to play a role in immunotherapy, but existing evidence to support its immuno-modulatory claims is insufficient. Four fractions from whole bee venom (BV) were separated using medium pressure liquid chromatography. Their ability to induce the production of cytokines TNFα, IL-1ß and IL-6 in phorbol-12-myristate-13-acetate (PMA)-treated U937 cells was assessed. The levels of the three cytokines produced by stimulation with the four fractions and crude BV without LPS were not significantly different from negative control values. However, co-stimulation of the cells with LPS and Fraction 4 (F-4) induced a 1.6-fold increase in TNF-α level (p < 0.05) compared to LPS alone. Likewise, LPS-induced IL-1ß production was significantly synergised in the presence of F-1 (nine-fold), F-2 (six-fold), F-3 (four-fold) and F-4 (two-fold) fractions, but was only slightly enhanced with crude BV (1.5-fold) relative to LPS. Furthermore, the LPS-stimulated production of IL-6 was not significantly increased in cells co-treated with F-2 and F-3, but the organic fraction (F-4) showed an inhibitory effect (p < 0.05) on IL-6 production. The latter was elucidated by NMR spectroscopy and found to contain(Z)-9-eicosen-1-ol. The effects observed with the purified BV fractions were more marked than those obtained with the crude sample.

An LCMS method for the assay of melittin in cosmetic formulations containing bee venom.

There is a growing interest in the potential of bee venom in cosmetics as a rejuvenating agent. Products currently on the market do not specify exactly their content of bee venom (BV). Therefore, we developed a method for the detection and quantification of melittin, as a marker of bee venom content, in selected commercial creams which contained BV according to their marketing claims, in order to gauge the relative quality of such formulations. A quantitative method was achieved following a rigorous extraction procedure involving sonication, liquid-liquid extraction and solid phase extraction since carryover of excipients was found to cause a rapid deterioration in the chromatographic performance. The method employed a standard additions approach using, as spiking standard, purified melittin isolated from bee venom and standardised by quantitative NMR. The aqueous extracts of the spiked creams were analysed by reversed phase LCMS on an LTQ Orbitrap mass spectrometer. The purity of the melittin spiking standard was determined to be 96.0%. The lowest measured mean melittin content in the creams was 3.19 ppm (±1.58 ppm 95% CI) while the highest was 37.21 ppm (±2.01 ppm 95% CI). The method showed adequate linearity (R (2) ≥ 0.98) and a recovery of 87.7-102.2% from a spiked blank cream. An assay precision of <20% RSD was achieved for all but one sample where the RSD value was 27.5%. The method was sensitive enough for use in routine assay of BV-containing cosmetic creams. Differences in the melittin content of the commercial products assayed were nearly tenfold.