Green Monkey Research Articles

Insights into the Geographical Origins of the Cabo Verde Green Monkey.

The green monkey Chlorocebus sabaeus, L. 1766, native to West Africa, was introduced to the Cabo Verde Archipelago in the 16th century. Historical sources suggest that, due to the importance of Cabo Verde as a commercial entrepôt in the Atlantic slave trade, establishing the precise place of origin of this introduced species is challenging. Non-invasive fecal samples were collected from feral and captive green monkey individuals in Cabo Verde. Two mitochondrial fragments, HVRI and cyt b, were used to confirm the taxonomic identification of the species and to tentatively determine the geographic origin of introduction to the archipelago from the African continent. By comparing the new sequences of this study to previously published ones, it was shown that Cabo Verde individuals have unique haplotypes in the HVRI, while also showing affinities to several populations from north-western coastal Africa in the cyt b, suggesting probable multiple sources of introduction and an undetermined most probable origin. The latter is consistent with historical information, but may also have resulted from solely using mtDNA as a genetic marker and the dispersal characteristics of the species. The limitations of the methodology are discussed and future directions of research are suggested.

Mutations in the F protein of the live-attenuated respiratory syncytial virus vaccine candidate ΔNS2/Δ1313/I1314L increase the stability of infectivity and content of prefusion F protein.

Respiratory syncytial virus (RSV) is the leading viral cause of bronchiolitis and pneumonia in infants and toddlers, but there currently is no licensed pediatric vaccine. A leading vaccine candidate that has been evaluated for intranasal immunization in a recently completed phase 1/2 clinical trial is an attenuated version of RSV strain A2 called RSV/ΔNS2/Δ1313/I1314L (hereafter called ΔNS2). ΔNS2 is attenuated by deletion of the interferon antagonist NS2 gene and introduction into the L polymerase protein gene of a codon deletion (Δ1313) that confers temperature-sensitivity and is stabilized by a missense mutation (I1314L). Previously, introduction of four amino acid changes derived from a second RSV strain "line 19" (I79M, K191R, T357K, N371Y) into the F protein of strain A2 increased the stability of infectivity and the proportion of F protein in the highly immunogenic pre-fusion (pre-F) conformation. In the present study, these four "line 19" assignments were introduced into the ΔNS2 candidate, creating ΔNS2-L19F-4M. During in vitro growth in Vero cells, ΔNS2-L19F-4M had growth kinetics and peak titer similar to the ΔNS2 parent. ΔNS2-L19F-4M exhibited an enhanced proportion of pre-F protein, with a ratio of pre-F/total F that was 4.5- to 5.0-fold higher than that of the ΔNS2 parent. The stability of infectivity during incubation at 4°C, 25°C, 32°C and 37°C was greater for ΔNS2-L19F-4M; for example, after 28 days at 32°C, its titer was 100-fold greater than ΔNS2. ΔNS2-L19F-4M exhibited similar levels of replication in human airway epithelial (HAE) cells as ΔNS2. The four "line 19" F mutations were genetically stable during 10 rounds of serial passage in Vero cells. In African green monkeys, ΔNS2-L19F-4M and ΔNS2 had similar growth kinetics, peak titer, and immunogenicity. These results suggest that ΔNS2-L19F-4M is an improved live attenuated vaccine candidate whose enhanced stability may simplify its manufacture, storage and distribution, which merits further evaluation in a clinical trial in humans.

DNA binding, potential anticancer, antioxidant and molecular docking simulations of some isonicotinohydrazide metal complexes with the impact of high energy γ-rays irradiation doses: Synthesis and structural characterization

This work focuses on the preparation and characterization of some VO(II), Co(II), Cu(II), Zn(II) and Hg(II) complexes (1–5) of N'-(2-aminobenzoyl)isonicotinohydrazide ligand (HL). The compounds were characterized using various analytical and spectral tools. The resulting powder samples of the as prepared complexes (3,5) were exposed to two gamma ray doses up to 150 and 200 kGy from highly-energetic 60Co γ-irradiation (hereafter referred to as (3F,5F) at dose value of 150 kGy and (3F*,5F*) at dose value of 200 kGy). The impact of γ-irradiation on the possible structural changes of these samples after gamma irradiation was studied. The ligand and as prepared Cu(II) (3), Hg(II) (5) and their γ-irradiated (3F,5F) and (3F*,5F*) samples were tested for free radical scavenging activity using the DPPH assay. The DNA cleavage activity using supercoiled plasmid DNA (calf thymus ct-DNA) was explored. The tested ligand (HL) and complexes (3,5) showed excellent DNA binding. Additionally, the in vitro antitumor activity of (HL) and its Cu(II) (3) and Hg(II) (5) complexes was scrutinized against the human breast carcinoma (MCF-7), human hepatocarcinoma (Hep-G2) cell lines and kidney epithelial normal cells extracted from African green monkey kidney (Vero) to investigate the potential safety towards the normal cells. Also, docking simulation was done on the active site of 4FM9 and 1H7K protein for liver cancer and breast cancer cell line, respectively and showed excellent results.

A measles-vectored vaccine candidate expressing prefusion-stabilized SARS-CoV-2 spike protein brought to phase I/II clinical trials: protection of African green monkeys from COVID-19 disease.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged at the end of 2019 and is responsible for the largest human pandemic in 100 years. Thirty-four vaccines are currently approved for use worldwide, and approximately 67% of the world population has received a complete primary series of one, yet countries are dealing with new waves of infections, variant viruses continue to emerge, and breakthrough infections are frequent secondary to waning immunity. Here, we evaluate a measles virus (MV)-vectored vaccine expressing a stabilized prefusion SARS-CoV-2 spike (S) protein (MV-ATU3-S2PΔF2A; V591) with demonstrated immunogenicity in mouse models (see companion article [J. Brunet, Z. Choucha, M. Gransagne, H. Tabbal, M.-W. Ku et al., J Virol 98:e01693-23, 2024, https://doi.org/10.1128/jvi.01693-23]) in an established African green monkey model of disease. Animals were vaccinated with V591 or the control vaccine (an equivalent MV-vectored vaccine with an irrelevant antigen) intramuscularly using a prime/boost schedule, followed by challenge with an early pandemic isolate of SARS-CoV-2 at 56 days post-vaccination. Pre-challenge, only V591-vaccinated animals developed S-specific antibodies that had virus-neutralizing activity as well as S-specific T cells. Following the challenge, V591-vaccinated animals had lower infectious virus and viral (v) RNA loads in mucosal secretions and stopped shedding virus in these secretions earlier. vRNA loads were lower in these animals in respiratory and gastrointestinal tract tissues at necropsy. This correlated with a lower disease burden in the lungs as quantified by PET/CT at early and late time points post-challenge and by pathological analysis at necropsy.IMPORTANCESevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for the largest human pandemic in 100 years. Even though vaccines are currently available, countries are dealing with new waves of infections, variant viruses continue to emerge, breakthrough infections are frequent, and vaccine hesitancy persists. This study uses a safe and effective measles vaccine as a platform for vaccination against SARS-CoV-2. The candidate vaccine was used to vaccinate African green monkeys (AGMs). All vaccinated AGMs developed robust antigen-specific immune responses. After challenge, these AGMs produced less virus in mucosal secretions, for a shorter period, and had a reduced disease burden in the lungs compared to control animals. At necropsy, lower levels of viral RNA were detected in tissue samples from vaccinated animals, and the lungs of these animals lacked the histologic hallmarks of SARS-CoV-2 disease observed exclusively in the control AGMs.

Investigation (In-Vitro) of Antiproliferative Properties of Pseudopediastrum boryanum (Turpin) E. Hegewald Extracts in Various Cancer Cell Lines

In this study, antiproliferative effects of Pseudopediastrum boryanum isolated and cultured from Tokat Yeşilırmak River benthic habitats were investigated with in-vitro experiments. Anticancer activity tests of algae extracts were tested in cancer cell lines using the BrdU cell proliferation ELISA method. Vero (African Green Monkey Kidney Tumour Cells) and HeLa (Human Uterine Carcinoma Cells) were examined in concentrations of 100, 250 and 500 µg/ml. As a result of antiproliferative tests, P. boryanum was found to have effective anti-cancer properties. P. boryanum extract concentrations of 250 and 500 µg/ml had a significant cytotoxic effect on Vero and HeLa cell lines after 24 hours. According to the results, P. boryanum is estimated to be a good option for quantitative pharmaceutical analyses and could be evaluated in the pharmaceutical industry.

Arthrospira maxima extract prevents and cures Zika virus infection: In vitro analysis with VERO cells

Zika virus (ZIKV) infections have increased in recent years due to the lack of vaccines, specific antiviral treatments, and the difficulty controlling Aedes vectors. This infection leads to serious neurological disorders, such as microcephaly and Guillain-Barré syndrome. Although higher plants are widely recognized as sources of bioactive compounds, there is relatively limited knowledge of such molecules in algae and microalgae. In this article, we evaluated Arthrospira maxima extract for its antiviral properties against ZIKV. We performed in vitro experiments in African green monkey kidney (VERO) cells. A. maxima extract showed a strong dose-dependent inhibition of ZIKV replication. Based on the half-maximal effective concentration, microalgal extract (1.65 ± 0.08 μg/mL) was more effective than the reference drug ribavirin (3.4 ± 0.8 μg/mL). The cytotoxicity assay showed that the microalgal extract was safer than ribavirin: The selectivity index of the extract was 5 times higher than ribavirin. The microalgal extract also had a protective and preventive role in inhibiting ZIKV binding to VERO cells. After pretreating these cells with 2.5, 5, 10, or 20 μg/mL of A. maxima extract for 2 h, the highest concentration had inhibited approximately 80 % viral adsorption. Finally, to investigate the direct impacts of A. maxima extract on the virus, we incubated ZIKV with 10 μg/mL of extract before infection (−2 or − 1 h), during infection, and at 2–8 h post infection. There was unprecedented virucidal activity of A. maxima algal extract against ZIKV at all times. In conclusion, A. maxima extract counteracts viral exertion and exerts virucidal activity to prevent ZIKV infection.

Molecular Detection and Genetic Diversity of Cytomegaloviruses and Lymphocryptoviruses in Free-Roaming and Captive African Green Monkeys (Chlorocebus sabaeus).

To date, limited information is available on cytomegalovirus (CMV) and lymphocryptovirus (LCV) from Chlorocebus monkeys. We report here high detection rates of herpesviruses in free-roaming African green monkeys (AGMs, Chlorocebus sabaeus) (26.4%, 23/87) and in captive AGMs (75%, 3/4) with respiratory disease on the Caribbean Island of St. Kitts. LCV (81.25%) was more prevalent than CMV (18.75%) in the AGMs. Applying a bigenic PCR approach (targeting DNA polymerase (DPOL) and glycoprotein B (gB) genes), long sequences were obtained from representative AGM CMV (KNA-SD6) and LCV (KNA-E4, -N6 and -R15) samples, and mixed LCV infections were identified in KNA-N6 and -R15. The nucleotide (nt) sequence (partial DPOL-intergenic region-partial gB) and partial DPOL- and gB-amino acid (aa) sequences of AGM CMV KNA-SD6 were closely related to Cytomegalovirus cercopithecinebeta5 isolates from grivet monkeys, whilst those of AGM LCV KNA-E4 and -N6 (and E4-like gB of KNA-R15) were more closely related to cognate sequences of erythrocebus patas LCV1 from patas monkey than other LCVs, corroborating the concept of cospeciation in the evolution of CMV/LCV. On the other hand, the partial DPOL aa sequence of KNA-R15, and additional gB sequences (N6-gB-2 and R15-gB-2) from samples KNA-N6 and -R15 (respectively) appeared to be distinct from those of Old World monkey LCVs, indicating LCV evolutionary patterns that were not synchronous with those of host species. The present study is the first to report the molecular prevalence and genetic diversity of CMV/LCV from free-roaming/wild and captive AGMs, and is the first report on analysis of CMV nt/deduced aa sequences from AGMs and LCV gB sequences from Chlorocebus monkeys.

Evaluation of a Combined Live Attenuated Vaccine against Lumpy Skin Disease, Contagious Bovine Pleuropneumonia and Rift Valley Fever.

The use of effective vaccines is among the most important strategies for the prevention and progressive control of transboundary infectious animal diseases. However, the use of vaccine is often impeded by the cost, a lack of cold chains and other factors. In resource-limited countries in Africa, one approach to improve coverage and reduce cost is to vaccinate against multiple diseases using combined vaccines. Therefore, the objective of this study was to evaluate a combined vaccine for the prevention and control of Lumpy Skin Disease (LSD), Contagious Bovine Pleuropneumonia (CBPP) and Rift Valley fever (RVF). The LSD and CBPP were formulated as a combined vaccine, and the RVF was formulated separately as live attenuated vaccines. These consisted of a Mycoplasma MmmSC T1/44 strain that was propagated in Hayflick-modified medium, RVF virus vaccine, C13T strain prepared in African green monkey cells (Vero), and the LSDV Neethling vaccine strain prepared in primary testis cells. The vaccines were tested for safety via the subcutaneous route in both young calves and pregnant heifers with no side effect, abortion or teratogenicity. The vaccination of calves induced seroconversions for all three vaccines starting from day 7 post-vaccination (PV), with rates of 50% for LSD, 70% for CBPP and 100% for RVF, or rates similar to those obtained with monovalent vaccines. The challenge of cattle vaccinated with the LSD/CBPP and the RVF vaccine afforded full protection against virulent strains of LSDV and RVFV. A satisfactory level of protection against a CBPP challenge was observed, with 50% of protection at 6 months and 81% at 13 months PV. A mass vaccination trial was performed in four regions of Burkina Faso that confirmed safety and specific antibody responses induced by the vaccines. The multivalent LSD/CBPP+RVF vaccine provides a novel and beneficial approach to the control of the three diseases through one intervention and, therefore, reduces the cost and improves vaccination coverage.

The impact of pumpkin seed-derived silver nanoparticles on corrosion and cytotoxicity: a molecular docking study of the simulated AgNPs

ABSTRACT Green-synthesized nanoparticles from pumpkin (Cucurbita pepo L.) seed extracts are economical and eco-friendly. Silver nanoparticles (AgNPs) and their selective cytotoxicity towards HCT116 and African Green Monkey Kidney, Vero cells were investigated. Chemical fingerprinting, heat stability, and 2D-images of nanoparticle size and morphology were determined using Fourier-transform infrared spectroscopy (FTIR), Thermogravimetric Analysis (TGA), Transmission Electron Microscopy (TEM), and Scanning electron microscopy-energy dispersive X-ray (SEM-EDX) on AgNPs. UV-vis examination shows surface plasmons in the wide peak at 417 nm, indicating polydisperse nanoparticles. Small silver nanoparticles (AgNPs) below 2 µm demonstrated a rod-like form and a tendency to agglomerate. SEM-EDX element analysis and fingerprinting confirmed the material as AgNPs. TEM indicated that the nanoparticles were generally spherical or ellipsoidal, equally dispersed, and averaged 26.08 nm in diameter with negligible aggregation. The AgNPs are also stable at a temperature of 220°C, indicating that the green material is quite robust at 150°C to 400°C. According to cytotoxic studies, AgNPs are toxic to cancer cells (HCT 116 cells), however they have no effect on Vero cells. AgNPs and tubulin (TUB) domain active sites have a significant affinity, according to molecular docking analysis. In an electrochemical investigation, biogenic AgNPs effectively prevented mild steel from corroding in a 1.0 M HCl solution.

A vaccine targeting antigen-presenting cells through CD40 induces protective immunity against Nipah disease

Nipah virus (NiV) has been recently ranked by the World Health Organization as being among the top eight emerging pathogens likely to cause major epidemics, whereas no therapeutics or vaccines have yet been approved. We report a method to deliver immunogenic epitopes from NiV through the targeting of the CD40 receptor of antigen-presenting cells by fusing a selected humanized anti-CD40 monoclonal antibody to the Nipah glycoprotein with conserved NiV fusion and nucleocapsid peptides. In the African green monkey model, CD40.NiV induces specific immunoglobulin A (IgA) and IgG as well as cross-neutralizing responses against circulating NiV strains and Hendra virus and Tcell responses. Challenge experiments using a NiV-B strain demonstrate the high protective efficacy of the vaccine, with all vaccinated animals surviving and showing no significant clinical signs or virus replication, suggesting that the CD40.NiV vaccine conferred sterilizing immunity. Overall, results obtained with the CD40.NiV vaccine are highly promising in terms of the breadth and efficacy against NiV.

New thiazol-pyridazine derivatives as antimicrobial and antiviral candidates: Synthesis, and application

In this manuscript, we are motivated to investigate the reaction site-selectivity for the hydrazo thiazole derivatives (6a–c) with different types of active methylene groups such as malononitrile, and ethyl cyanoacetate. Based on their structural investigations and spectrum data, the results of these reactions have been established to be iminopyridazines (7a–c) and 6-oxopyridazine derivatives (8a–c). We tested the ability of the newly synthesized pyridazine derivatives to inhibit the microbes and COVID-19 proteins. Human coronavirus 229E (HCoV-229E) was used to investigate the antiviral efficacy of prepared compounds. Green monkey kidney (Vero-E6) cell lines were used to investigate MTT and cytopathic effect (CPE). The new 6-oxopyridazine derivatives (8a–c) revealed significant inhibitory efficacy and were capable of inhibiting the human coronavirus 229E. Moreover, the antimicrobial result showed that compounds iminopyridazine (7c) followed by iminopyridazine (7a) followed by iminopyridazine (7b) exhibited excellent antimicrobial properties toward all utilized strains, usually greater than that of common reference drugs, with MIC values ranging from 13 to 21 ppm, from 9 to 14 ppm, and from 8 to 19 ppm whereas, the remaining substances appeared to be promising effective. Structure-activity relationship (SAR) revealed that pyridazine scaffolds containing NH group, as well as substituted electron withdrawal group (Cl) in para-position for benzene ring attributed to thiazole moiety have the best activity. The current study successfully illustrated the possible application of heterocyclic derivatives with pyridazie nucleus including thiazole ring as the main compound in the development of dual antiviral (COVID-19) and antibacterial pharmaceuticals in the future.

Qualitative Phytochemical Screening, Fatty Acid Profile and Biological Studies of the Bark of <i>Mallotus nudiflorus</i> (Pitali) Plant

In the present study, the Mallotus nudiflorus (L.) plant has been taken to determine the in-vitro analysis to find out the therapeutic value. The bioassays of the raw extract of methanol of bark of M. nudiflorus and by Kupchan’s extraction method collecting n-Hexane (HEX), Dichloromethane (DCM), Chloroform (CHCl3), Ethylacetate (EA) and Aqueous (AQ) fractions were scrutinized to find out its therapeutic value. The findings of phytochemical screening of the methanol extract of barks revealed the presence of several secondary metabolites. By using the GC-FID method the result showed that M. nudiflorus contained four bound fatty acids and four free fatty acids. EA fraction had the maximum phenolic content among all the fractions at (133.67±0.99) mg of GAE/g where the Folin- Ciocalteu reagent was used as an oxidizing agent. The antioxidant activity was measured in terms of its ability to scavenge free radicals (DPPH assay). Among all extractives, the greatest ability to scavenge for free radicals was shown by EA extract with an IС50 value (12.08±0.15) μg/ml. In the toxicity of brine shrimp test, the HEX fraction had the maximum toxicity with an LC50 value of (0.12±0.01) μg/ml. Cell cytotoxicity was observed for sample CHCl3 and EA on both the Vero (kidney epithelial cells taken from an African green monkey) and HeLa (a human cervical cancer cell) cell line. All the fractions were subjected to in vitro microbial screening, which revealed that DCM, CHCl3, and EA fractions showed growth inhibition, particularly against various Gram-positive and Gram-negative bacteria by disc diffusion method. The maximum zone of inhibition in the antimicrobial activity was produced by CHCl3 fraction against Staphylococcus aureus (17 mm). The results of in vitro experiments have demonstrated that the extracts from the barks of M. nudiflorus have great potential for medicinal uses and might be studied for further chemical exploration.

The 8-hydroxyquinoline derivative, clioquinol, is an alpha-1 adrenoceptor antagonist

Clioquinol (5-chloro-7-iodo-8-hydroxyquinoline) is an antimicrobial agent whose actions as a zinc or copper ionophore and an iron chelator revived the interest in similar compounds for the treatment of fungal and bacterial infections, neurodegeneration and cancer. Recently, we reported zinc ionophores, including clioquinol, cause vasorelaxation in isolated arteries through mechanisms that involve sensory nerves, endothelium and vascular smooth muscle. Here, we report that clioquinol also uniquely acts as a competitive alpha-1 (α1) adrenoceptor antagonist. We employed ex vivo functional vascular contraction and pharmacological techniques in rat isolated mesenteric arteries, receptor binding assays using stabilized solubilized α1 receptor variants, or wild-type human α1-adrenoceptors transfected in COS-7 cells (African green monkey kidney fibroblast-like cells), and molecular dynamics homology modelling based on the recently published α1A adrenoceptor cryo-EM and α1B crystal structures. At higher concentrations, all ionophores including clioquinol cause a non-competitive antagonism of agonist-mediated contraction due to intracellular zinc delivery, as reported previously. However, at lower concentration ranges, clioquinol has an additional mechanism of competitively inhibiting α1-adrenoceptors that contributes to decreasing vascular contractility. Molecular dynamic simulation showed that clioquinol binds stably to the orthosteric binding site (Asp106) of the receptor, confirming the structural basis for competitive α1-adrenoceptor antagonism by clioquinol.

Efficient gene delivery by multifunctional star poly (β-amino ester)s into difficult-to-transfect macrophages for M1 polarization

Gene delivery to macrophages holds great promise for cancer immunotherapy. However, traditional gene delivery methods exhibit low transfection efficiency in macrophages. The star-shaped topological structure of polymers is known to encapsulate genes inside their cores, thereby facilitating sustained release of the genetic material. Herein, combining the structural advantages of star polymers and the transfection advantages of poly (β-amino ester)s (PAEs), we developed a novel linear oligomer grafting-onto strategy to synthesize a library of multi-terminal star structured PAEs (SPAEs), and evaluated their gene delivery efficiency in various tissue cells. The transfection with human hepatocellular carcinoma cells (HepG2, HCC-LM3 cells and MHCC-97H cells), rat normal liver cells (BRL-3 A cells), human ovarian cancer cells (A2780 cells), African green monkey kidney cells (Vero cells), human cervical cancer cells (HeLa cells), human chondrosarcoma cells (SW1353 cells), and difficult-to-transfect human epidermal keratinocytes (HaCaT cells) and normal human fibroblast cells (NHF cells) showed that SPAEs exhibited superior transfection profile. The GFP transfection efficiency of top-performing SPAEs in HeLa cells (96.1%) was 2.1-fold, and 3.2-fold higher compared to jetPEI and Lipo3000, respectively, indicating that the star-shaped topological structure can significantly enhance the transfection efficiency of PAEs. More importantly, the top-performing SPAEs could efficiently deliver Nod2 DNA to difficult-to-transfect RAW264.7 macrophages, with a high transfection efficiency of 33.9%, which could promote macrophage M1 polarization and enhanced CD8+ T cell response in co-incubation experiments. This work advances gene therapy by targeting difficult-to-transfect macrophages and remodeling the tumor immune microenvironment.

Preliminary in vitro evaluation of the cytoprotective effects of clarified açaí (Euterpe oleracea MART.) pulp on African green monkey kidney cells (VERO) treated with the antifungal fluconazole

Preliminary in vitro evaluation of the cytoprotective effects of clarified açaí (Euterpe oleracea MART.) pulp on African green monkey kidney cells (VERO) treated with the antifungal fluconazole

Bioflocculation of pollutants in wastewater using flocculant derived from Providencia huaxiensis OR794369.1

BackgroundWater pollution has become a major environmental and health concern due to increasing population and industrialisation. Microbial flocculants are promising agents for treatment of contaminated water owing to their effectiveness, eco-friendliness, and high biosafety levels. In this study, culture conditions of Providencia huaxiensis OR794369.1 were optimised and its bioflocculant was extracted, characterised and used to treat wastewater.ResultsThe maximum flocculating activity of 92% and yield of 3.5 g/L were obtained when cultivation conditions were: 3% inoculum size, starch, casein, initial pH of 6, cultivation temperature of 30 oC and 72 h of fermentation. The bioflocculant is an amorphous glycoprotein biomolecule with 37.5% carbohydrates, 27.9% protein, and 34.6% uronic acids. It is composed of hydroxyl, amino, alkanes, carboxylic acid and amines groups as its main functional structures. It was found to be safe to use as it demonstrated non-cytotoxic effects on bovine dermis and African green monkey kidney cells, illustrating median inhibitory concentration (IC50) values of 180 and > 500 µg/mL on both cell lines, respectively. It demonstrated the removal efficiencies of 90% on chemical oxygen demand (COD), 97% on biological oxygen demand (BOD) and 72% on Sulphur on coal mine wastewater. It also revealed the reduction efficacies of 98% (COD) and 92% (BOD) and 70% on Sulphur on domestic wastewater.ConclusionThe bioflocculant was effective in reducing pollutants and thus, illustrated potential to be used in wastewater treatment process as an alternative.

Drug repurposing platform for deciphering the druggable SARS-CoV-2 interactome

The coronavirus disease 2019 (COVID-19) pandemic has heavily challenged the global healthcare system. Despite the vaccination programs, the new virus variants are circulating. Further research is required for understanding of the biology of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and for discovery of therapeutic agents against the virus. Here, we took advantage of drug repurposing to identify if existing drugs could inhibit SARS-CoV-2 infection. We established an open high throughput platform for in vitro screening of drugs against SARS-CoV-2 infection. We screened ∼1000 drugs for their ability to inhibit SARS-CoV-2–induced cell death in the African green monkey kidney cell line (Vero-E6), analyzed how the hit compounds affect the viral N (nucleocapsid) protein expression in human cell lines using high-content microscopic imaging and analysis, determined the hit drug targets in silico, and assessed their ability to cause phospholipidosis, which can interfere with the viral replication. Duvelisib was found by in silico interaction assay as a potential drug targeting virus–host protein interactions. The predicted interaction between PARP1 and S protein, affected by Duvelisib, was further validated by immunoprecipitation. Our results represent a rapidly applicable platform for drug repurposing and evaluation of the new emerging viruses’ responses to the drugs. Further in silico studies help us to discover the druggable host pathways involved in the infectious cycle of SARS-CoV-2.

Mammalian Cell Cytotoxicity, Antibacterial Activity and the Properties of Methylenebis(Hydroxybenzoic Acid) and Its Related Zinc(II) Complex

Formaldehyde, sulfuric acid and salicylic acid were combined to create a 3,3′-methylenebis(2-hydroxybenzoic acid) (MHB) ligand, which was subsequently permitted to bind with zinc(II) ions. The ligand and its zinc(II) complex (Zn–MHB) have been described by a combination of elemental analyses, spectral analyses (UV–Vis, IR, MS and NMR), XRD, TEM, as well as TGA measurement. The ligand has been suggested to coordinate to the zinc center in a tetradentate manner forming the binuclear tetrahedral complex. An X-ray analysis indicated a considerable difference between MHB (crystalline) and Zn–MHB (amorphous). The UV–Vis spectra were used to determine the optical properties such as bandgap, refractive index, optical conductivity and penetration depth. The possibility of employing the samples for optoelectronic applications was indicated from the band gap values which underlie the range of semiconductors. TEM revealed the spherical shapes and mutation of ligand particles into the nano-scale by complexation. The antimicrobial potential of the MHB towards Gram-positive and Gram-negative bacterial growths has been investigated. The results suggested that it would be possible to employ MHB to prevent bacterial development, particularly that of salmonella typhimurium. The cytotoxicity of the MHB was assessed against two types of mammalian cells: VERO (the kidney of an African green monkey) and HFB4 (human skin melanocytes). Lower sensitivity was observed in VERO cells.

Bioguided antiplasmodial isolation of six new labdane-type diterpenoids from the stem bark of Croton sylvaticus Hochst. Ex. Krauss

The CH2Cl2-MeOH (1:1, v/v) extract of Croton sylvaticus stem bark selectively exhibited good antiplasmodial activity during preliminary screening against the multidrug-resistant (Dd2) and chloroquine sensitive (3D7) strains of Plasmodium falciparum with IC50 values of 10.57 and 18.47 μg/mL, respectively. After fractionation, the fractions exhibited moderate to good activity with IC50 values ranging from 4.31 to 25.49 μg/mL. The purification of these fractions led to the isolation of six new labdane-type diterpenoids named sylvacrotonins AF (16), alongside seven known compounds (713). Their structures were determined based on the analysis of their NMR and MS data. The isolated compounds were assessed in vitro for their antiplasmodial activity against Pf3D7 and PfDd2. The new labdane-type diterpenoids showed moderate antiplasmodial activity against the two P. falciparum strains, with compound 6 being the most active, with IC50 values of 11.26 and 21.80 μg/mL against PfDd2 and Pf3D7, respectively. Cytotoxic assays were performed on African green monkey kidney Vero cells to ensure that the obtained activity was specific to the parasites. Interestingly, all the tested extracts, fractions, and isolated compounds showed acceptable selectivity (SI > 10) against both Plasmodium strains, except fractions CSB-E (SI = 3.93) and CSB-C (SI = 9.22) against Pf3D7. Globally, with resistance indexes (RIs) lower than 1, all the tested samples were found to be more active against multidrug-resistant (PfDd2) than sensitive (Pf3D7) strains. Furthermore, all the isolated compounds exhibited cell cytotoxicity in the normal Vero cell line. The results from this study confirm the use of C. sylvaticus in malaria-endemic countries for its management and suggest that further lead-optimization studies on hit compounds could drive to the identification of potential lead molecules for antimalarial control and eradication.

In vitro biological activities of Combretum molle R.Br. ex G. Don (Combretaceae) against mastitis-causing organisms

Bovine mastitis, caused mostly by Staphylococcus species, has gained global importance owing to the increasing prevalence of multi-drug resistant bacteria, which comprise a risk to food security and public health. In this study, the antibacterial, anti-biofilm and quorum quenching activities of six leaf extracts of Combretum molle R.Br. ex G. Don prepared using different solvents were assessed against clinical isolates of Staphylococcus aureus and two Staphylococcus ATCC strains (S. aureus ATCC 29213 and S. epidermidis ATCC 35984). The antioxidant and anti-inflammatory activity and cytotoxicity of the extracts was also evaluated. The antibacterial and anti-biofilm potential of the extracts was determined via serial microdilution and crystal violet and INT assays. Quorum quenching activity was ascertained via inhibition of violacein production in Chromobacterium violaceum ATCC 12472. The antioxidant activity was determined using in vitro chemical assays. The 15-lipoxygenase enzyme inhibition and the nitric oxide inhibition assays were utilized to ascertain the anti-inflammatory activity of the extracts. The tetrazolium-based colorimetric (MTT) reduction assay was used to determine the cytotoxicity of the extracts against Vero African green monkey kidney cells. The antibacterial activity of the extracts was moderate to good, with minimum inhibitory concentration (MIC) values of 0.02–0.63 mg/mL. The extracts had promising biofilm inhibition (≥50 %) against four of the test strains. The effects of the extracts on the metabolic activity of the test strains showed ≥ 50 % inhibition in most of the test strains at the different test times. All extracts had moderate to excellent quorum quenching activity at different concentrations with an inhibition ranging between 32.81- 96.32 %. The methanol and cold water extracts had the best antioxidant activity against the 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and 2, 2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radicals with IC50 of 7.79 and 2.34 µg/mL, respectively. The hot water extract (AQH) had the best anti-inflammatory activity in terms of 15-lipoxygenase enzyme inhibition with 85.26 % inhibition, while the methanol extract had the best nitric oxide inhibition in LPS-induced RAW 264.7 macrophages with inhibition of 96.75 % at 100 µg/mL. Selectivity index values of the extracts were as high as 50. Combretum molle, therefore, offers promising potential as a source of antibiofilm and anti-quorum sensing agents with the ability to disrupt microbial virulence factors. Additionally, the extract had good antioxidant and anti-inflammatory properties, supporting further research into its use in preventing and treating bovine mastitis.