Phytochemical screening and antibacterial activity of Vernonia amygdalina (bitter leaf) on some selected bacterial isolates

Plants with medicinal value produce certain chemical elements known as phytochemicals that have antibacterial activity. The study was aimed at determining the antibacterial activity of Vernonia amygdalina against bacterial isolates using agar well diffusion method. In addition, the phytochemicals analysis of the extracts was also determined. The phytochemical analysis showed the presence of saponins, steroids, terpenoids, tannins, alkaloids, and flavonoids. The result of Vernonia amygdalina showed that the average zones of inhibitions observed against these bacterial ranges from 6-22mm. The highest zone is also exhibited against E. coli with average diameter of zone of inhibition of 22mm. At 100mg/ml concentration for Samonella, the zone of inhibition was recorded to be 21mm while at 12.5mg/ml there was no inhibition. At 25mg/ml and 12.5mg/ml, against Pseudomonas there was no inhibition. In other to further confirm the activity of these plant extracts, the minimum inhibitory concentration and minimum bactericidal concentration was determined and the result showed that the extract exerted good antibacterial activity on all the test organisms at different concentration. The result of minimum inhibitory concentration ranges from 10 to 12.5mg/ml and that of MBC ranges from 5 to 20mg/ml. It is worthy to note that MBC values is greater than that of minimum inhibitory concentration. The study provides insight into the antibacterial activities of the plant extracts and its use in the treatment of bacterial infections.

Hypoxia Increases Nitric Oxide-Dependent Inhibition of Angiogenic Growth

Nitric oxide (NO) is a proangiogenic factor acting through the soluble guanylate cyclase (sGC) pathway. However, angiogenic growth increases energy demand, which may be hampered by NO inhibition of cytochrome c oxidase (CcO). Then, NO activity would be the balanced result of sGC activation (pro-angiogenic) and CcO inhibition (anti-angiogenic). NO activity in a rat and eNOS−/− mice aortic ring angiogenic model and in a tube formation assay (human aortic endothelial cells) were analyzed in parallel with mitochondrial O2 consumption. Studies were performed with NO donor (DETA-NO), sGC inhibitor (ODQ), and NOS or nNOS inhibitors (L-NAME or SMTC, respectively). Experiments were performed under different O2 concentrations (0–21%). Key findings were: (i) eNOS-derived NO inhibits angiogenic growth by a mechanism independent on sGC pathway and related to inhibition of mitochondrial O2 consumption; (ii) NO inhibition of the angiogenic growth is more evident in hypoxic vessels; (iii) in the absence of eNOS-derived NO, the modulation of angiogenic growth, related to hypoxia, disappears. Therefore, NO, but not lower O2 levels, decreases the angiogenic response in hypoxia through competitive inhibition of CcO. This anti-angiogenic activity could be a promising target to impair pathological angiogenesis in hypoxic conditions, as it occurs in tumors or ischemic diseases.

Cross-reactivity of antibodies from non-hospitalized COVID-19 positive individuals against the native, B.1.351, B.1.617.2, and P.1 SARS-CoV-2 spike proteins

SARS-CoV-2 variants of concern (VOCs) have emerged worldwide, with implications on the spread of the pandemic. Characterizing the cross-reactivity of antibodies against these VOCs is necessary to understand the humoral response of non-hospitalized individuals previously infected with SARS-CoV-2, a population that remains understudied. Thirty-two SARS-CoV-2-positive (PCR-confirmed) and non-hospitalized Canadian adults were enrolled 14–21 days post-diagnosis in 2020, before the emergence of the B.1.351 (also known as Beta), B.1.617.2 (Delta) and P.1 (Gamma) VOCs. Sera were collected 4 and 16 weeks post-diagnosis. Antibody levels and pseudo-neutralization of the ectodomain of SARS-CoV-2 spike protein/human ACE-2 receptor interaction were analyzed with native, B.1.351, B.1.617.2 and P.1 variant spike proteins. Despite a lower response observed for the variant spike proteins, we report evidence of a sustained humoral response against native, B.1.351, B.1.617.2 and P.1 variant spike proteins among non-hospitalized Canadian adults. Furthermore, this response inhibited the interaction between the spike proteins from the different VOCs and ACE-2 receptor for ≥ 16 weeks post-diagnosis, except for individuals aged 18–49 years who showed no inhibition of the interaction between B.1.617.1 or B.1.617.2 spike and ACE-2. Interestingly, the affinity (KD) measured between the spike proteins (native, B.1.351, B.1.617.2 and P.1) and antibodies elicited in sera of infected and vaccinated (BNT162b2 and ChAdOx1 nCoV-19) individuals was invariant. Relative to sera from vaccine-naïve (and previously infected) individuals, sera from vaccinated individuals had higher antibody levels (as measured with label-free SPR) and more efficiently inhibited the spike–ACE-2 interactions, even among individuals aged 18–49 years, showing the effectiveness of vaccination.

New chalcone derivatives as potential antimicrobial and antioxidant agent

Seven chalcone derivatives were synthesized by the Claisen-Schmidt condensation. The structures of the compounds were confirmed by spectral data (Ultraviolet/visible, infrared, nuclear magnetic resonance and mass spectroscopy). The compounds were tested for their in silico and in vitro antimicrobial and antioxidant activities. The molecular docking assessments showed that all the compounds exhibited good binding affinity with the target microorganism proteins but, compounds 6e and 6g showed better binding affinity compared with the standards. The antimicrobial test revealed that all the compounds screened were active against Staphylococcus aureus and Bacillus subtilis and had minimum inhibitory concentrations (MIC) between 0.4 and 0.6 mg/mL. Compounds 6a, 6c and 6d had moderate activities on Salmonella typhi. Compounds 6b and 6c had moderate activity on Escherichia coli. Compound 6c had moderate activity on Aspergillus niger while compounds 6a and 6e had poor activity. All the compounds except compound 6e had no inhibition against Pseudomonas aeruginosa. The in-vitro antioxidant activity was assessed using ethylenediaminetetraacetate (EDTA) as the standard. Compounds 6c, 6e and 6g gave excellent inhibitory activity better than the standard. Compound 6a gave good activity at 500 μg/mL and 1000 μg/mL concentrations but, below the standard at 250 μg/mL and no inhibition at 125 μg/mL. Compound 6d had good inhibition at 500 μg/mL and 1000 μg/mL but, no inhibition at 125 μg/mL and 250 μg/mL. Compound 6b was found to be inactive in all the concentrations. Absorption, distribution, metabolism and excretion properties of the compounds were assessed using SwissADME. The results of lead likeness showed that compound 6e is a lead-like molecule.

Anti-inflammatory activity of raw and processed stingless bee honey

The anti-inflammatory activity of raw and processed Kelulut stingless bee honey was investigated for its ability to inhibit nitric oxide (NO) production in lipopolysaccharidestimulated RAW 264.7 cells. Raw honey was optimally processed by thermal processing and thermosonication at 90°C. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) cell viability assay showed that Kelulut honey from 7.8 to 500 µg/mL was not cytotoxic to RAW 264.7 cells as it resulted in at least 80% viable cells after 24 hr. Both raw and processed honey from 10 to 300 µg/mL displayed an increase and decrease in NO concentrations, suggesting a mixed effect of NO inhibition and enhancement. Maximum NO inhibition of 17.5% was recorded from 20 µg/mL of thermally processed honey while the highest NO enhancement of 7.8% was from 10 µg/mL of thermosonicated honey. The NO effects were independent of honey concentration and processing techniques, suggesting its potential robustness in medicinal properties as part of the diet.

Comparing the efficacy of Neem extract, Aloe vera juice, and 0.2% Chlorhexidine against dental unit waterline pathogens. an In-vitro study

Introduction: Nosocomial infections are caused by the retrograde spread of viral, bacterial and fungal pathogens from the hospital environment to the patients. Despite the efforts to avoid cross-infection in the dental office using sterilized instruments, individual protection equipment, and disinfection procedures, there is an increased risk of cross-infection through dental units, since one dental chair is used to treat many patients. Materials and methods: Seven strains of micro-organisms- Streptococcus, Lactobacillus, Pseudomonas aeruginosa, Klebsiella, Enterococcus, Staphylococcus aureus, Candida albicans, E. coli, and Legionella were taken. Neem extract, Aloe Vera Juice, and 0.2% chlorhexidine were tested against all these organisms. Sterile discs were incorporated with an equal amount of prepared formulations using a micropipette. Then these discs were placed equidistant to each other following which these plates were incubated for 24 hours. Results: Zone of inhibition was higher in chlorhexidine 12mm, 22mm, 11mm, 8mm, 18mm, 12mm, 11mm, 14mm and 15 mm followed by neem extract 7mm, no inhibition, 10mm, 7mm, 11mm, 7mm, 11mm and 9 mm and no inhibition by Aloe Vera juice against Streptococcus, Lactobacillus, Pseudomonas aeruginosa, Klebsiella, Enterococcus, Staphylococcus Aureus, Candida albicans, E.coli and Legionella respectively. Conclusion: Results from this study have shown that chlorhexidine was most effective against dental unit waterline pathogens followed by neem extract and least by Aloe Vera juice.

Are Hydrogen Peroxide, Hypochlorite and Peracetic Acid Potential Endocrine Disruptors?

Arylsulfatase and β-glucuronidase are two important enzymes in human, which play important role on dynamic equilibrium of steroidal estrogens. This work probably for the first time reported that hydrogen peroxide (H2O2), hypochlorite and peracetic acid (PAA) could effectively inhibit the activities of arylsulfatase and/or β-glucuronidase. The 50 percent of inhibitions (IC50) of H2O2, and PAA on arylsulfatase were found to be 142.90±9.00, 91.83±10.01, and 43.46±2.92 μM, respectively. The corresponding respective IC50 of hypochlorite and PAA on β-glucuronidase were 704.90±41.40 and 23.26±0.82 μM, while H2O2 showed no inhibition on β-glucuronidase. It was further revealed that the inhibition of hypochlorite on both arylsulfatase and β-glucuronidase was irreversible. On the contrary, the inhibition by H2O2 and PAA was reversible. Moreover, it was found that the inhibitions of arylsulfatase and/or β-glucuronidase by these three chemicals were pH-dependent, among which the inhibition by H2O2 was competitive and non-competitive for PAA. In general, H2O2 and hypochlorite can be endogenously produced in human, which suggested that the two compounds are potential endocrine disruption compounds (EDCs) as they can cause endocrine disruption via inhibition of arylsulfatase and β-glucuronidase. This work further indicated that any agent that can induce production of H2O2 or hypochlorite in human is potential EDC, which explains why some EDCs with very weak or no estrogenic potency can cause endocrine disruption that confirmed in epidemiological studies.

Multiple circulating alkaloids and saponins from intravenous Kang-Ai injection inhibit human cytochrome P450 and UDP-glucuronosyltransferase isozymes: potential drug-drug interactions

Background: Kang-Ai injection is widely used as an adjuvant therapy drug for many cancers, leukopenia, and chronic hepatitis B. Circulating alkaloids and saponins are believed to be responsible for therapeutic effects. However, their pharmacokinetics (PK) and excretion in vivo and the risk of drug-drug interactions (DDI) through inhibiting human cytochrome P450 (CYP) and UDP-glucuronosyltransferase (UGT) enzymes remain unclear. Methods: PK and excretion of circulating compounds were investigated in rats using a validated ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS) method. Further, the inhibitory effects of nine major compounds against eleven CYP and UGT isozymes were assayed using well-accepted specific substrate for each enzyme. Results: After dosing, 9 alkaloids were found with Cmax and t1/2 values of 0.17-422.70 μmol/L and 1.78-4.33 h, respectively. Additionally, 28 saponins exhibited considerable systemic exposure with t1/2 values of 0.63-7.22 h, whereas other trace saponins could be negligible or undetected. Besides, over 90% of alkaloids were excreted through hepatobiliary and renal excretion. Likewise, astragalosides and protopanaxatriol (PPT) type ginsenosides also involved in hepatobiliary and/or renal excretion. Protopanaxadiol (PPD) type ginsenosides were mainly excreted to urine. Furthermore, PPD-type ginsenosides were extensively bound (fu-plasma approximately 1%), whereas astragalosides and PPT-type ginsenosides displayed fu-plasma values of 12.35% and 60.23-87.36%, respectively. Moreover, matrine, oxymatrine, astragaloside IV, ginsenoside Rg1, ginsenoside Re, ginsenoside Rd, ginsenoside Rc, and ginsenoside Rb1 exhibited no inhibition or weak inhibition against several common CYP and UGT enzymes IC50 values between 8.81 and 92.21 μM. Through kinetic modeling, their inhibition mechanisms towards those CYP and UGT isozymes were explored with obtained Ki values. In vitro-in vivo extrapolation showed the inhibition of systemic clearance for CYP or UGT substrates seemed impossible due to [I]/Ki no more than 0.1.Conclusions: We summarized the PK behaviors, excretion characteristics and protein binding rates of circulating alkaloids, astragalosides and ginsenosides after intravenous Kang-Ai injection. Furthermore, weak inhibition or no inhibition towards these CYP and UGT activities could not trigger harmful DDI when Kang-Ai injection is co-administered with clinical drugs primarily cleared by these CYP or UGT isozymes.

Multiple circulating alkaloids and saponins from intravenous Kang-Ai injection inhibit human cytochrome P450 and UDP-glucuronosyltransferase isozymes: potential drug-drug interactions

Background: Kang-Ai injection is widely used as an adjuvant therapy drug for many cancers, leukopenia, and chronic hepatitis B. Circulating alkaloids and saponins are believed to be responsible for therapeutic effects. However, their pharmacokinetics (PK) and excretion in vivo and the risk of drug-drug interactions (DDI) through inhibiting human cytochrome P450 (CYP) and UDP-glucuronosyltransferase (UGT) enzymes remain unclear. Methods: PK and excretion of circulating compounds were investigated in rats using a validated ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS) method. Further, the inhibitory effects of nine major compounds against eleven CYP and UGT isozymes were assayed using well-accepted specific substrate for each enzyme. Results: After dosing, 9 alkaloids were found with Cmax and t1/2 values of 0.17-422.70 μmol/L and 1.78-4.33 h, respectively. Additionally, 28 saponins exhibited considerable systemic exposure with t1/2 values of 0.63-7.22 h, whereas other trace saponins could be negligible or undetected. Besides, over 90% of alkaloids were excreted through hepatobiliary and renal excretion. Likewise, astragalosides and protopanaxatriol (PPT) type ginsenosides also involved in hepatobiliary and/or renal excretion. Protopanaxadiol (PPD) type ginsenosides were mainly excreted to urine. Furthermore, PPD-type ginsenosides were extensively bound (fu-plasma approximately 1%), whereas astragalosides and PPT-type ginsenosides displayed fu-plasma values of 12.35% and 60.23-87.36%, respectively. Moreover, matrine, oxymatrine, astragaloside IV, ginsenoside Rg1, ginsenoside Re, ginsenoside Rd, ginsenoside Rc, and ginsenoside Rb1 exhibited no inhibition or weak inhibition against several common CYP and UGT enzymes IC50 values between 8.81 and 92.21 μM. Through kinetic modeling, their inhibition mechanisms towards those CYP and UGT isozymes were explored with obtained Ki values. In vitro-in vivo extrapolation showed the inhibition of systemic clearance for CYP or UGT substrates seemed impossible due to [I]/Ki no more than 0.1.Conclusions: We summarized the PK behaviors, excretion characteristics and protein binding rates of circulating alkaloids, astragalosides and ginsenosides after intravenous Kang-Ai injection. Furthermore, weak inhibition or no inhibition towards these CYP and UGT activities could not trigger harmful DDI when Kang-Ai injection is co-administered with clinical drugs primarily cleared by these CYP or UGT isozymes.

Optimization of Enzymatic Conditions of Sturgeon Muscles and Their Anti-Inflammatory Potential

The objective of this study was to investigate the effects of different enzymolysis conditions on the NO inhibition rate and DH (degree of hydrolysis) of sturgeon hydrolysates (SH) prepared by Alcalase. The NO inhibition rate of 60.23% was attained under the optimum enzymolysis conditions as follows: pH 9.0, enzymolysis time of 4.92 h, enzymolysis temperature of 55°C, solid/liquid ratio of 1 : 20, and enzyme additive amount of 7674.22 U/g protein, which was well matched with the predicted value 61.44% of the Box–Behnken design model. After the ultrafiltration of SH, SH-3 (SH < 3 kDa) could significantly decrease the levels of NO and proinflammatory cytokine level IL-6. Also, we found that the obtained SH-3 contained good properties of emulsification and possessed good WHC and OHC. SH-3 demonstrated appreciable antioxidant potential on DPPH and ABTS radical scavenging activities. These results suggested that SH-3 derived from sturgeon muscles could potentially be used as a promising ingredient against inflammatory and oxidative stress-associated diseases.