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Mosquito-borne viral diseases like chikungunya and dengue infections can cause severe illness and have become major public health concerns. Chikungunya virus (CHIKV) and dengue virus (DENV) infections share similar primary clinical manifestations and are transmitted by the same vector. Thus, the probability of their coinfection gets increased with more severe clinical complications in the patients. The present study was undertaken to elucidate the common human interacting partners of CHIKV and DENV proteins during coinfection. The viral–host protein–protein interactome was constructed using Cytoscape. Subsequently, significant host interactors were identified during coinfection. The network analysis elucidated 57 human proteins interacting with both CHIKV and DENV, represented as hub-bottlenecks. The functional and biological analyses of the 40 hub-bottlenecks revealed that they are associated with phosphoinositide 3-kinases (PI3K)/AKT, p53 signaling pathways, regulation of cell cycle and apoptosis during coinfection. Moreover, the molecular docking analysis uncovered the tight and robust binding of selected hub-bottlenecks with CHIKV/DENV proteins. Additionally, 23 hub-bottlenecks were predicted as druggable candidates that could be targeted to eradicate the host–viral interactions. The elucidated common host binding partners during DENV and CHIKV coinfection as well as indicated approved drugs can support the therapeutics development. [ABSTRACT FROM AUTHOR]

Glioblastoma is one of the most debilitating forms of brain tumour. It accounts for 17% of all the brain tumours and has resulted in 251,329 deaths in 2020 itself. Several studies performed on patient profiling have revealed that many genes tend to be overexpressed or mutated in many GBM patients and hamper the signalling and growth pathways. Additionally, there is a requirement to explore supplementary or alternative treatment that can aid in overcoming the resistance caused by standard care treatment (Temozolomide with radiation). The preliminary analysis on inhibitory potential of certain phytocompounds on the crucial genes involved in GBM has been studied in silico. The network of the genes critical for GBM has been analysed through several plugins of Cytoscape software like BiNGO and MCODE based on factors such as degree, closeness and betweenness. The network analysis gave out a cluster of 97 interactions with 16 interacting proteins. ERBB4 emerged as the seed protein indicating its crucial role in pathways. The BiNGO plugin gave out the gene ontology data describing the functions of the genes of the cluster. Additionally, the properties of ligands were studied by the PharmaGist server and 3D patterns of shared features by all or most input ligands of selected phytocompound were analysed and aligned. Upon docking the seed protein (ERBB4) individually with the best combination of aligned ligands yielded bromelain and EGCG as the compounds with best binding affinity of -9.8 and -9.2 kcal/mol respectively. This study has provided an important lead for further in vitro study to analyse the effect of combination of phytocompounds on the critical signalling pathways of GBM. [ABSTRACT FROM AUTHOR]

Purpose: Nanoemulsions (NEs) of polyphenon 60 (P60) and cranberry (NE I) and P60 and curcumin (NE II) were prepared with the aim to enhance anti-bacterial potential and to understand the mechanism of anti-bacterial action of the encapsulated compounds. Methods: To evaluate the antibacterial potential of the developed NE, microtiter biofilm formation assay was performed. The cytotoxicity analysis was done to assess the toxicity profile of the NEs. Further antibacterial analysis against uropathogenic strains was performed to check the developed NEs were effective against these strains. Results: In microtiter dish biofilm formation assay, both NE formulations inhibited the growth more effectively (Av. % inhibition ~84%) as compared to corresponding aqueous solution (Av. % inhibition ~64%) and placebo (Av. % inhibition ~59%) at their respective minimum inhibitory concentration (MIC) values. Cytotoxicity analysis using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT assay) showed that the formulations were nontoxic to Vero cells. The antibacterial studies against uropathogenic resistant strains also showed that NEs effectively inhibited the growth of bacterial strains. Conclusion: From different studies it was concluded that both the NE’s were able to inhibit bacterial strains and could be further used for the treatment of urinary tract infection (UTI). The antibacterial activity of developed NEs showed that these could be used as alternative therapies for the treatment of UTI. [ABSTRACT FROM AUTHOR]

Aim and Objectives: In the present investigation dual agents i.e., hesperidin and diazepam loaded polymeric nanoparticles (NPs) were formulated by nanoprecipitation method and optimized using three-level factorial design. Methods: The developed NPs were optimized keeping poly (lactic-co-glycolic) acid (PLGA), poloxamer amount as independent process variable and z-average, percentage drug entrapment as a dependent response. The optimized NP was subjected to in vitro drug release study to investigate drug release mechanism from NP. Cell viability assay was performed on Vero cell line to confi rm the safety of NP. Results: Drug loaded NP showed z-average in the range of 189-307 d.nm with percentage drug entrapment for diazepam and hesperidin 62-89% and 68-92%, respectively. In vitro drug release studies showed controlled drug release behavior was observed from polymeric NP across dialysis membrane compared to aqueous drug solution. Cell viability assay showed drug dependent cytotoxicity on Vero cell line, however, polymeric NP showed less cytotoxicity compared with aqueous drug solution. [ABSTRACT FROM AUTHOR]

Objective: The aim of the current study was to isolate and identify the strain having broad spectrum antibacterial and antibiofilm activity and characterize its antimicrobial substance. Methods: Lactic acid bacteria isolated from the raw milk of Indian goat species Babri and Sirohi (Capra aegagrus hircus) were screened for antibacterial activity against Staphylococcus epidermidis, Micrococccus luteus, Pseudomonas fluorescens and Escherichia coli. The selected strains were also analysed for anti-biofilm activity on S. epidermidis. The potential bacterial strain was characterized at molecular level by 16S rDNA sequencing. The antimicrobial substance produced by P. acidilactici was precipitated by 90% ammonium sulphate and subsequently ultra filtered. It was analysed for size, stability under various pH, temperature, chemicals and enzymatic conditions. Its mechanism of killing was determined by ML35p assay and scanning electron microscopy (SEM). Results: The 16S rDNA sequence of the potential isolate, possessing broad spectrum antibacterial activity and antibiofilm activity against S. epidermidis, was deposited to Genbank under accession no. KP671843 and was identified as Pediococcus acidilactici. The partially purified bacteriocin ran at approximately 3500 Da on tris-tricine SDS-PAGE and showed antibacterial activity against S. epidermidis by In-gel assay. Partially purified bacteriocin was stable at wide pH and high temperature and also to the treatment of lipase, amylase, catalase and various detergents/organic solvents. However, it was sensitive to proteolytic enzymes indicating its proteinaceous nature. Results of membrane permeability assay revealed that the bacteriocin targets bacterial cell membrane as its bactericidal action. Conclusion: The bacteriocin from P. acidilactici has the potential to be used as broad spectrum antibacterial and antibiofilm agent. [ABSTRACT FROM AUTHOR]

Objective: The aim of this study was to purify and characterize bacteriocin from the soil isolate Bacillus subtilis GAS101, and to determine its antimicrobial as well as antibiofilm potential. The purified bacteriocin was further analyzed and evaluated for mammalian cell cytotoxicity and the possible mode of action. Material and Methods: Bacteriocin from B. subtilis GAS101 (an animal husbandry soil isolate) was partially purified and checked for antimicrobial and antibiofilm activity against gram-positive and gram-negative bacteria. The molecular weight of bacteriocin was determined using tricine SDS-PAGE gel. The stability of bacteriocin was investigated at various temperatures and pH levels, and its sensitivity towards 8 enzymes and 6 chemicals was determined. Cytotoxicity analysis was performed on a Vero cell line by a tetrazolium dye-based assay. Scanning electron microscopy (SEM) of bacteriocin-treated bacteria was carried out to determine the possible mode of action. Results: Bacteriocin from B. subtilis GAS101 was a potential inhibitor of both the indicator organisms (Staphylococcus epidermidis and Escherichia coli), and had a molecular weight of approximately 6.5 kDa. An in situ gel assay showed a zone of inhibition corresponding to the estimated protein band size. Bacteriocin was stable and showed antibacterial activity in broad ranges of temperature (30–121°C) and pH (2–12). It was sensitive to 4 proteolytic enzymes, which indicated its proteinaceous nature. Bacteriocin showed > 70% cell viability on the mammalian Vero cell line. SEM depicted that the bacteriocin was able to disrupt the bacterial cell membrane as its probable mode of action. Conclusion: Thermostable and pH-tolerant bacteriocin from B. subtilis GAS101, of about 6.5 kDa, showed broad-spectrum antimicrobial and antibiofilm activity. [ABSTRACT FROM AUTHOR]

The aim of the present study was to develop a nanoemulsion-based delivery system containing green tea catechins called polyphenon 60 and ciprofloxacin for intravaginal delivery to treat urinary tract infection. Polyphenon 60 and ciprofloxacin were encapsulated in a single nanoemulsion system prepared using ultrasonication technique. The nanoemulsion was characterized by determining particle size, zeta potential, morphological structure and estimating in vitro release and antibacterial efficacy. To determine the in vivo pharmacokinetic parameters and intravaginal transportation of nanoemulsion in Sprague Dawley rats, gamma scintigraphy and biodistribution study was conducted with technetium pertechnetate-labelled nanoemulsion. The preliminary antibacterial investigation showed synergy between these compounds with FICindex of 0.42. The developed formulation showed zeta potential of +55.3 mV and globule size of 151.7 nm, with polydispersity index of 0.196. The percent in vitro release for polyphenon 60 at the end of 7 h was 94.8±0.9, whereas for ciprofloxacin it was 75.1±0.15 in simulated vaginal media. Antibacterial activity evaluation against extended spectrum beta lactamase and metallo beta lactamase strains revealed that nanoemulsions containing 4 and 10 mg/ml each of polyphenon 60 and ciprofloxacin effectively inhibited the growth of bacterial strains. In biodistribution study, the percent radiolabelled drug per gram was found to be 3.50±0.26 and 3.81±0.30 in kidney and urinary bladder, respectively at 3 h. From these findings it could be concluded that the developed polyphenon 60+ciprofloxacin nanoemulsion showed antibacterial activity against Escherichia coli and was transported efficiently to the target organs through vaginal mucosa. [ABSTRACT FROM AUTHOR]

Objective: The aim of this study was to purify and characterize bacteriocin from the soil isolate Bacillus subtilis GAS101, and to determine its antimicrobial as well as antibiofilm potential. The purified bacteriocin was further analyzed and evaluated for mammalian cell cytotoxicity and the possible mode of action.Material and Methods: Bacteriocin from B. subtilis GAS101 (an animal husbandry soil isolate) was partially purified and checked for antimicrobial and antibiofilm activity against gram-positive and gram-negative bacteria. The molecular weight of bacteriocin was determined using tricine SDS-PAGE gel. The stability of bacteriocin was investigated at various temperatures and pH levels, and its sensitivity towards 8 enzymes and 6 chemicals was determined. Cytotoxicity analysis was performed on a Vero cell line by a tetrazolium dye-based assay. Scanning electron microscopy (SEM) of bacteriocin-treated bacteria was carried out to determine the possible mode of action.Results: Bacteriocin from B. subtilis GAS101 was a potential inhibitor of both the indicator organisms (Staphylococcus epidermidis and Escherichia coli), and had a molecular weight of approximately 6.5 kDa. An in situ gel assay showed a zone of inhibition corresponding to the estimated protein band size. Bacteriocin was stable and showed antibacterial activity in broad ranges of temperature (30-121°C) and pH (2-12). It was sensitive to 4 proteolytic enzymes, which indicated its proteinaceous nature. Bacteriocin showed > 70% cell viability on the mammalian Vero cell line. SEM depicted that the bacteriocin was able to disrupt the bacterial cell membrane as its probable mode of action.Conclusion: Thermostable and pH-tolerant bacteriocin from B. subtilis GAS101, of about 6.5 kDa, showed broad-spectrum antimicrobial and antibiofilm activity. [ABSTRACT FROM AUTHOR]

Purpose: A nanoemulsion based gel containing Polyphenon 60 (P60) and cranberry (CRB) has been developed to deliver via intravaginal route for the treatment of urinary tract infection. Methods: Polyphenon 60 and cranberry were loaded in a single nanoemulsion gel (NBG) by ultra-sonication method and characterized for particle size, rheological properties, in vitro release and growth curve analysis. P60+CRB NBG were radiolabelled using technetium pertechnetate (99mTc) to perform in vivo pharmacokinetic studies in animals. Results: The finalized NE had a droplet size of 58±1 nm. In vitro release of 90.92 ± 0.6% in 8 hr for P60 and 99.39 ± 0.5% in 6 hr for CRB was observed in simulated vaginal fluid. Growth curve of E. coli indicated the inhibitory action of nanoemulsion based gel at the fifth hour of inoculation. Gamma scintigraphy studies on female Sprague-Dawley rats showed transport of nanoemulsion based gel from the vaginal cavity into the systemic circulation. Further, biodistribution studies with radiolabelled P60+CRB NBG showed significant higher uptake of radiolabelled actives by kidney (3.20±0.16) and urinary bladder (3.64±0.29), when administered intravaginally. Conclusion: The findings suggested 99mTc-P60+CRB NBG can potentially be transported through vaginal cavity and reach the target organs and showed effective distribution in organs affected in urinary tract infection [ABSTRACT FROM AUTHOR]

Staphylococcus epidermidis has emerged as the main causative agent for graft-related and nosocomial infections. Rampant use of antibiotics and biofilm formed by the organism results in poor penetration of the drug and further aggravates the antibiotic resistance, emphasizing an urgent need to explore alternative treatment modalities. Antimicrobial peptides (AMPs), produced as effector molecules of the innate immunity of living organisms, have therapeutic potential that can be used to inhibit the growth of microbes. In addition, the susceptibility of a microbe to become resistant to an AMP is relatively low. The AMPs are amphipathic peptides of 12-100 residues, which have broad-spectrum activity against microbes. There are scattered reports of AMPs listed against S. epidermidis and there is an urgent need to systematically study the AMPs. Various natural AMPs as well as synthetic peptides have been investigated against S. epidermidis. These peptides have been shown to inhibit both planktonic culture and S. epidermidis biofilm effectively. The multiple modes of action in killing the organism minimize the chances for the development of resistance. This review focused on various natural and synthetic peptides that demonstrate activity against S. epidermidis. [ABSTRACT FROM AUTHOR]

One of the most important rapidly emerging mosquito-borne alphavirus is Chikungunya virus (CHIKV). There is a necessity to develop anti-CHIKV therapeutics, as neither antiviral drug nor vaccines have been licensed yet. Several CHIKV proteins are being studied worldwide, but non-structural protein 3 (nsP3) has been less explored. This protein consists of three domains: macrodomain, alphavirus unique domain (AUD) and hypervariable region (HVR). The proline-rich regions of HVR contain SRC homology 3 (SH3)-binding domain which is essential for its functionality. Interaction of these motifs with host amphiphysin protein is crucial for viral RNA replication. Restricting the interactions of HVR could lead to inhibition of viral life cycle. Therefore, the present study focuses on purification of HVR protein and its structural and functional assay for therapeutic intervention in future use. In order to obtain purified protein, HVR region was amplified from TOPO clones of nsP3 of IND-06-Guj strain and cloned into expression vector. Expression and solubilization of the protein were optimized at various conditions of salt, detergent and imidazole before purification. The soluble recombinant HVR (His-HVR) protein was purified using affinity chromatography. Purified protein was analyzed for structural studies and functional assays. Circular dichroism of His-HVR protein was performed for structural study, and it was observed that it consists of mostly random coils. For functional assay, co-pull down of His-HVR protein was performed with endogenous amphiphysin-I protein of N2a cells and was analyzed using Western blotting. This purified protein obtained could be used as a potential target reagent for novel therapeutic interventions in the future. [ABSTRACT FROM AUTHOR]