Effect of Christmas Melon (Laganaria Breviflorus) extract on toxigenic Mycoflora Isolated from Stored Unpolished Rice sold in major Markets in Abeokuta, Nigeria

Study on toxigenic mycoflora and potential mitigation effect of Christmas Melon (Laganaria Breviflorus) extract in unpolished rice sold in Abeokuta Ogun state of Nigeria was carried out. Unpolished rice gotten from markets in Abeokuta were aseptically transported to the laboratory, serial dilution to reduce the fungal load was carried out and were plated on Potato Dextrose Agar (PDA) and Methyl Red Dessicated Coconut Agar (MRDCA) respectively. Microscopy, macroscopy, toxigenicity test and inhibition studies with the peeled and unpeeled fruit of Laganaria breviflorus fermented for seven days was carried out. Results reveal the predominance of Aspergillus as the major genera, specifically, A. niger, A.flavus, A. parasiticus, A. fumigatus, A. terreus, A. nidulans. Other fungi genera isolated include Penicillium, F`usarium, Mucor, Alternaria and Rhizopus . Of the 11 fungi genera isolated, 9 were toxigenic of which the zones of inhibition of unpeeled whole fruit extract of Laganaria breviflorus range from (3 - 28mm) where A. nidulans shows the highest susceptibility to the whole fruit extract of Laganaria breviflorus while the zone of inhibition of peeled fruit extract of Laganaria breviflorus ranges from (3 - 22mm) where A. parasiticus, Fusarium specie and P.chrysogenum showed the highest susceptibility . As the day progresses the zone of inhibition becomes wider. Unpeeled LB extract exhibited more zones of inhibition than the peeled LB extract. Laganaria breviflorus fruit extracts in the study demonstrates a potential in reducing toxigenic fungi, consequently a means to reducing mycotoxins in staple foods in Nigeria.

CYP-450 Mediated Herb-Drug Interactions of Ashwagandha and AYUSH-64 Used in COVID-19 Integrative Management: A Case Example of Remdesivir

AimWithania somnifera Dunal (WS), known as Ashwagandha and AYUSH-64, a polyherbal formulation are repurposed for the management of COVID-19. The extensive use of these botanicals as home remedy along with other drugs for managing multifarious disease conditions is increasing over nations. This raises high chances of herb-drug interactions (HDIs) which may be beneficial, harmful, or even fatal. Therefore, current study aimed to explore the CYP mediated herb-drug interactions (HDIs) of Ashwagandha and AYUSH-64 along with case example of remdesivir to harness the best of these HDIs for integrative management of COVID-19Materials and MethodsThe aqueous extract of Ashwagandha and AYUSH-64 were characterized by LC-MS/MS. The in-silico pharmacokinetic (ADME) parameters were studied by using ADME tool. The in-vitro CYP-450 (CYP3A4, 2C8, 2D6) inhibition studies of WS and AYUSH-64 alone and in combination with a remdesivir were carried out resembling clinically scenario.ResultsTotal of 11 and 24 phytoconstituents were identified from the aqueous extract of Aswagandha and AYUSH-64. The in-silico ADME studies revealed that most of the phytoconstituents showed good oral bioavailability, drug likeliness, GI affinity and some of them displayed CYP-450 inhibitory activity. The in-vitro CYP-450 studies of remdesivir showed moderate inhibitory effect on CYP3A4, 2C8, 2D6. The aqueous extract of Aswagandha did not show any inhibitory activity towards all the studied CYP’s alone and in combination with remdesivir (IC50 >100µg/ml). Whereas, AYUSH-64 also followed the same trend but showed moderate inhibitory effect on CYP2C8 (IC50 <100µg/ml).ConclusionAswagandha did not exhibit HDIs with the CYP3A4, CYP2C8 and CYP2D6 thereby seem to be safe to co-administer with respective substrates. Whereas, AYUSH-64 only showed moderate HDIs towards CYP2C8 substrate among studied CYP enzymes. Caution is therefore warranted for prescribing AYUSH 64 along with CYP2C8 substrate drugs.

A intermediate (Aint) subgroup with warm anti-A1 antibody: A case report

A intermediate (Aint) subtypes exhibit characteristics intermediate between A1 and A2. Plasma from Aint individuals contains different enzyme, UDP-GalNAc: fucosylgalactoside-a-3-N-acetylgalactosaminyl transferase, which is different from the enzyme in A1 and A2 plasma. We encountered the case of a 54-year-old female (having pneumonia and chronic kidney disease) for pre-transfusion testing. On routine grouping, we encountered group discrepancies. On testing, anti-A gave 4+, anti-B-0, anti-A1 lectin-2+, anti-H lectin, and anti-AB antisera gave 4+ reactions. Reverse grouping gave 4+ with B cells, 2+ at room temperature with A cells, and 4+ and 1+ at 37°C and 4°C. Saliva inhibition studies showed A and H substances. It was typed as an Aint group with warm anti-A1 antibody. It’s the 1st time ever we encountered Aint case with a warm type anti-A1 antibody. Here, O group packed red cells are the suitable blood units to transfuse.

Synthesis and Inhibition Activity Study of Triazinyl-Substituted Amino(alkyl)-benzenesulfonamide Conjugates with Polar and Hydrophobic Amino Acids as Inhibitors of Human Carbonic Anhydrases I, II, IV, IX, and XII

Primary sulfonamide derivatives with various heterocycles represent the most widespread group of potential human carbonic anhydrase (hCA) inhibitors with high affinity and selectivity towards specific isozymes from the hCA family. In this work, new 4-aminomethyl- and aminoethyl-benzenesulfonamide derivatives with 1,3,5-triazine disubstituted with a pair of identical amino acids, possessing a polar (Ser, Thr, Asn, Gln) and non-polar (Ala, Tyr, Trp) side chain, have been synthesized. The optimized synthetic, purification, and isolation procedures provided several pronounced benefits such as a short reaction time (in sodium bicarbonate aqueous medium), satisfactory yields for the majority of new products (20.6–91.8%, average 60.4%), an effective, well defined semi-preparative RP-C18 liquid chromatography (LC) isolation of desired products with a high purity (>97%), as well as preservation of green chemistry principles. These newly synthesized conjugates, plus their 4-aminobenzenesulfonamide analogues prepared previously, have been investigated in in vitro inhibition studies towards hCA I, II, IV and tumor-associated isozymes IX and XII. The experimental results revealed the strongest inhibition of hCA XII with low nanomolar inhibitory constants (Kis) for the derivatives with amino acids possessing non-polar side chains (7.5–9.6 nM). Various derivatives were also promising for some other isozymes.

Contributions of UDP-Glucuronosyltransferases to Human Hepatic and Intestinal Metabolism of Ticagrelor and Inhibition of UGTs and Cytochrome P450 Enzymes by Ticagrelor and its Glucuronidated Metabolite

Ticagrelor is the first reversibly binding, direct-acting, oral P2Y12 receptor inhibitor. The contribution of UDP-glucuronosyltransferases (UGTs) enzymes to the metabolism of ticagrelor to its glucuronide conjugation, ticagrelor-O-glucuronide, in human liver microsomes (HLM) and human intestinal microsomes (HIM), was well characterized in the current study. The inhibition potential of human major UGTs by ticagrelor and ticagrelor-O-glucuronide was explored. The inhibitory effects of ticagrelor-O-glucuronide on cytochrome P450s (CYPs) enzymes were investigated as well. Ticagrelor glucuronidation exhibits substrate inhibition kinetics in both HLM and HIM with apparent Km values of 5.65 and 2.52 μM, Vmax values of 8.03 and 0.90 pmol min−1·mg protein−1, Ksi values of 1,343.0 and 292.9 respectively. The in vitro intrinsic clearances (Vmax/Km) for ticagrelor glucuronidation by HLM and HIM were 1.42 and 0.36 μl min−1·mg protein−1, respectively. Study with recombinant human UGTs suggested that multiple UGT isoforms including UGT1A9, UGT1A7, UGT1A3, UGT1A4, UGT1A1, UGT2B7 and UGT1A8 are involved in the conversion of ticagrelor to ticagrelor-O-glucuronide with UGT1A9 showing highest catalytic activity. The results were further supported by the inhibition studies on ticagrelor glucuronidation with typical UGT inhibitors in pooled HLM and HIM. Little or no inhibition of UGT1A1, UGT1A3, UGT1A4, UGT1A6, UGT1A9 and UGT2B7 by ticagrelor and ticagrelor-O-glucuronide was noted. Ticagrelor-O-glucuronide also exhibited limited inhibitory effects toward CYP2C8, CYP2D6 and CYP3A4. In contrast, ticagrelor-O-glucuronide weakly inhibited CYP2B6, CYP2C9 and CYP2C19 activity with apparent IC50 values of 45.0, 20.0 and 18.8 μM, respectively. The potential of ticagrelor-O-glucuronide to cause drug-drug interactions warrant further study.

Raman Characterization of Fungal DHN and DOPA Melanin Biosynthesis Pathways

Fungal melanins represent a resource for important breakthroughs in industry and medicine, but the characterization of their composition, synthesis, and structure is not well understood. Raman spectroscopy is a powerful tool for the elucidation of molecular composition and structure. In this work, we characterize the Raman spectra of wild-type Aspergillus fumigatus and Cryptococcus neoformans and their melanin biosynthetic mutants and provide a rough “map” of the DHN (A. fumigatus) and DOPA (C. neoformans) melanin biosynthetic pathways. We compare this map to the Raman spectral data of Aspergillus nidulans wild-type and melanin biosynthetic mutants obtained from a previous study. We find that the fully polymerized A. nidulans melanin cannot be classified according to the DOPA pathway; nor can it be solely classified according to the DHN pathway, consistent with mutational analysis and chemical inhibition studies. Our approach points the way forward for an increased understanding of, and methodology for, investigating fungal melanins.

Iron-Zinc Co-Doped Titania Nanocomposite: Photocatalytic and Photobiocidal Potential in Combination with Molecular Docking Studies

In the current research study, iron-zinc co-doped TiO2 was reported as an energy efficient material for the degradation of DIPA and inactivation of E. coli and S. aureus under visible light irradiation. In addition, molecular docking simulation was performed to provide further insight into possible targets for inhibiting bacterial development. The synthesized nanocomposites were screened and optimized for different synthesis and reaction parameters. The physicochemical properties of the synthesized nanocomposites were evaluated through different characterization techniques. The wet impregnation (WI) approach was among the most successful methods for the synthesis of Fe-Zn-TiO2 nanocomposite (NC) utilizing anatase titanium. Moreover, 66.5% (60 min reaction time) and 100% (190 min reaction time) chemical oxygen demand (COD) removal was obtained through optimized NC, i.e., 0.1Fe-0.4Zn metal composition and 300 °C calcination temperature. The energy consumption for the best NC was 457.40 KW h m−3. Moreover, 0.1Fe-0.4Zn-TiO2-300 was more efficient against S. aureus compared to E. coli with 100% reduction in 90 min of visible light irradiations. Furthermore, 0.1Fe-0.4Zn-TiO2-300 NC showed that the binding score for best docked conformation was −5.72 kcal mol−1 against β-lactamase from E. coli and −3.46 kcal mol−1 from S. aureus. The studies suggested the Fe-Zn in combination with TiO2 to be a possible inhibitor of β-lactamase that can be further tested in enzyme inhibition studies.

Cinnamic acid inhibits cell viability, invasion, and glycolysis in primary endometrial stromal cells by suppressing NF-κB-induced transcription of PKM2

Background: Endometriosis is a painful disorder characterized by the growth of endometrial tissue outside the uterine cavity. Here, we investigated the effects of the cinnamic acid isolated from the Chinese medicinal plant Cinnamomum cassia Presl on primary endometrial stromal cells. Methods: Immunohistochemistry was used to examine protein expression and cell purity. Quantitative RT-PCR was conducted to assess mRNA expression, and Western blot was performed to determine protein level. Cell viability was assessed using cell counting kit-8 (CCK-8) assay. Glycolysis and mitochondrial function were evaluated by measuring the extracellular acidification rate (ECAR) and the oxygen consumption rate (OCR) of cells, respectively. Lastly, plasmid transfection and inhibitor treatment were used for overexpression and inhibition studies. Results: Cinnamic acid inhibited cell viability and cell invasion, as well as decreased ECAR and OCR, in primary endometrial stromal cells. Cinnamic acid suppressed the effects of PKM2 overexpression, and inhibition of PKM2 using Compound 3k mimicked the effects of cinnamic acid. Treatment with Compound 3k and cinnamic acid did not lead to additive effects, but rather displayed effects similar to those of Compound 3k alone, suggesting that cinnamic acid elicits its effects on primary endometrial stromal cells by targeting PKM2. Conclusions: Our study identified cinnamic acid as a novel compound from Cinnamomum cassia Presl that displays potent effects on primary endometrial stromal cell viability, invasion, and glycolysis, suggesting its potential use for endometriosis treatment.