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Glioblastoma multiforme (GBM) is a grade IV, lethal, and the most common type of brain tumor. GBM can acquire resistance to temozolomide (TMZ) recommended for its treatment. Naringenin (NAG), a flavonoid generally found in grapefruit, has antioxidant, anti-proliferative, and anti-inflammatory properties. It has been reported that phytochemicals can reduce resistance and improve the efficacy of a chemo-resistant drug. The combinatorial effect of TMZ and NAG on cell proliferation was evaluated using 3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT) assay, and the apoptosis in the U87MG and LN229 GBM cells were evaluated by change in fluorescence intensity. The effect of NAG and TMZ on anchorage-independent single-cell colony formation and cell migration was investigated. NAG and TMZ demonstrated enhanced cytotoxic effects on U87MG and LN229 cell lines. The combination index value being less than one indicated the synergistic action of the two drugs in restricting the growth of the cells. The NAG and TMZ together resulted in higher fluorescence intensity as compared to the alone drug. Further, the study showed a marked reduction in the migration of the cells and the formation of a single cell colony. Supplemental data for this article is available online at https://doi.org/10.1080/01635581.2021.1952438 [ABSTRACT FROM AUTHOR]

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]

Despite rapid advances in the human healthcare, the infection caused by certain viruses results in high morbidity and mortality accentuate the importance for development of new antivirals. The existing antiviral drugs are limited, due to their inadequate response, increased rate of resistance and several adverse side effects. Therefore, one of the newly emerging field "peptide-based therapeutics" against viruses is being explored and seems promising. Over the last few years, a lot of scientific effort has been made for the identification of novel and potential peptide-based therapeutics using various advanced technologies. Consequently, there are more than 60 approved peptide drugs available for sale in the market of United States, Europe, Japan, and some Asian countries. Moreover, the number of peptide drugs undergoing the clinical trials is rising gradually year by year. The peptide-based antiviral therapeutics have been approved for the Human immunodeficiency virus (HIV), Influenza virus and Hepatitis virus (B and C). This review enlightens the various peptide sources and the different approaches that have contributed to the search of potential antiviral peptides. These include computational approaches, natural and biological sources (library based high throughput screening) for the identification of lead peptide molecules against their target. Further the applications of few advanced techniques based on combinatorial chemistry and molecular biology have been illustrated to measure the binding parameters such as affinity and kinetics of the screened interacting partners. The employment of these advanced techniques can contribute to investigate antiviral peptide therapeutics for emerging infections. [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]

• AUD is a multifunctional domain of nsP3 essential for Chikungunya virus life cycle. • The human interactors of CHIKV AUD were identified utilizing yeast two hybrid system. • SNAPIN and N4BP2L2 interaction with CHIKV AUD was validated by pull down assay. • Key residues involved in SNAPIN and N4BP2L2 association with AUD were predicted. The life-threatening re-emerged chikungunya virus (CHIKV) can cause an epidemic outbreak and still has no vaccine available so far. Alphavirus unique domain (AUD) of CHIKV nsP3 is a multifunctional domain that remains conserved among alphaviruses and is critical for CHIKV replication. The understanding of AUD-host protein-protein interactions and their association with the cellular processes concerning CHIKV infection are not well studied. In the current study, the protein-protein interactions of AUD and its human host were elucidated by screening of universal human cDNA library using yeast two-hybrid system. The chosen interactions were further validated by GST pull-down assay, and their network mapping was analyzed. The study revealed that the identified interactors are linked with the vesicle trafficking and transcription corepressor activities. Further, the interfacial residues of interactions between viral and host proteins were predicted, which will further provide the new platform to develop novel antivirals. [ABSTRACT FROM AUTHOR]

Solanum nigrum is used in various traditional medical systems for antiproliferative, antiinflammatory, antiseizure and hepatoprotective activities. We have evaluated organic solvent and aqueous extracts obtained from berries of Solanum nigrum for antiproliferative activity on leukemic cell lines, Jurkat and HL-60 (Human promyelocytic leukemia cells). The cell viability after the treatment with Solanum nigrum extract was measured by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay. Results indicated increased cytotoxicity with increasing extract concentrations. Comparative analysis indicated that 50% inhibitory concentration value of methanol extract is the lowest on both cell lines. [ABSTRACT FROM AUTHOR]

Systemic lupus erythematosus (SLE) is an autoimmune disease associated with hyperactive innate and adaptive immune systems that cause dermatological, cardiovascular, renal, and neuropsychiatric problems in patients. SLE's multifactorial nature and complex pathogenesis present significant challenges in its clinical classification. In addition, unpredictable treatment responses in patients emphasize the need for highly specific and sensitive SLE biomarkers that can assist in understanding the exact pathogenesis and, thereby, lead to the identification of novel therapeutic targets. Recent studies on microRNA (miRNA), a non-coding region involved in the regulation of gene expression, indicate its importance in the development of the immune system and thus in the pathogenesis of various autoimmune disorders such as SLE. miRNAs are fascinating biomarker prospects for SLE categorization and disease monitoring owing to their small size and high stability. In this paper, we have discussed the involvement of a wide range of miRNAs in the regulation of SLE inflammation and how their modulation can be a potential therapeutic approach. [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]

Baclofen and Lamotrigine via PLGA nanoparticles were developed for nose-to-brain delivery for the treatment of Neuropathic pain. Nanoparticles were prepared using the modified nano-precipitation method. The prepared NPs were characterised and further in vitro and in vivo studies were performed. The Bcf-Ltg-PLGA-NPs were ∼177.7 nm with >75%(w/w) drugs encapsulated. In vitro dissolution studies suggested zero-order release profiles following the Korsmeyer-Peppas model. In vitro cytotoxicity and staining studies on mammalian cells showed dose dependant cytotoxicity where nanoparticles were significantly less toxic (>95% cell-viability). ELISA studies on RAW-macrophages showed Bcf-Ltg-PLGA-NPs as a potential pro-inflammatory-cytokines inhibitor. In vivo gamma-scintigraphy studies on rats showed intra-nasal administration of 99mTc-Bcf-Ltg-PLGA-NPs showed Cmax 3.6%/g at Tmax = 1.5h with DTE% as 191.23% and DTP% = 38.61% in brain. Pharmacodynamics evaluations on C57BL/6J mice showed a significant reduction in licks/bites during inflammation-induced phase II pain. The findings concluded that the combination of these drugs into a single nanoparticle-based formulation has potential for pain management. Baclofen and Lamotrigine loaded PLGA nanoparticles were prepared with a size of 177.7nm, PDI 0.057 and Zeta Potential −15.8 mV In vitro cell lines based studies showed dose dependant cytotoxicity and Bcf-Ltg-PLGA-NPs were found to be pro-inflammatory cytokines inhibitors In vivo Pharmacokinetic studies showed Cmax 3.6%/g at Tmax = 1.5 h with Drug Targeting Efficiency 191.23% and Drug Target Organ Transport 38.61% in the brain for prepared nanoparticles In vivo pharmacodynamics studies showed a significant reduction in licks/bites during inflammation-induced phase II pain [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]

In this work, poly (D,L-lactide-co-glycolic acid) (PLGA) nanoparticles of baclofen (Bcf-PLGA-NPs) were developed and optimized using nanoprecipitation method. The average particle size of the Bcf-PLGA-NP was found to be 124.8 nm, polydispersity index of 0.225, and zeta potential was found to be in the range of -20.4 mV. In vitro dissolution studies showed that Bcf was released from PLGA NPs in a sustained manner from 50% release in 2.5 hours to 80%-85% in 24 hours. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay on Neuro-2a neuroblastoma cell line showed comparably low cytotoxicity of Bcf-PLGA-NPs as compared with aqueous solution of Bcf at reported Cmax values of the drug. To explore the nose-to-brain pathway, in vivo studies were carried out in Sprague-Dawley rats by radiolabeling of Bcf with technetium-99m (99mTc). Gamma scintigraphy images of the rats that were administered through intranasal (i.n.) route showed the maximum uptake of radiolabeled NPs from nose to brain at 3 hours as compared with the rats administered with NPs intravenously and orally. To assess the Bcf concentration in brain and blood, biodistribution studies were performed and following i.n. route the NPs were dispersed in brain (3.5%/g) and blood (3%/g) at 3 hours, and these observations were in agreement with the gamma scintigrams. Hence, from the results it was suggested that the developed PLGA NPs could serve as a potential carrier for the Bcf in the treatment of neuropathic pain. [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]

Bacteriocins are antimicrobial peptides which are ribosomally synthesized by mainly all bacterial species. LABs (lactic acid bacteria) are a diverse group of bacteria that include around 20 genera of various species. Though LABs have a tremendous potential for production of anti-microbial peptides, this group of bacteria is still underexplored for bacteriocins. To study the diversity among bacteriocin encoding clusters and the putative bacteriocin precursors, genome mining was performed on 20 different species of LAB not reported to be bacteriocin producers. The phylogenetic tree of gyrB, rpoB, and 16S rRNA were constructed using MEGA6 software to analyze the diversity among strains. Putative bacteriocins operons identified were found to be diverse and were further characterized on the basis of physiochemical properties and the secondary structure. The presence of at least two cysteine residues in most of the observed putative bacteriocins leads to disulphide bond formation and provide stability. Our data suggests that LABs are prolific source of low molecular weight non modified peptides. [ABSTRACT FROM AUTHOR]

Staphylococcus epidermidisis reported to be the main causative agent of nosocomial infections. It has become increasingly difficult to treat this micro-organism because of the emergence of new antibiotic-resistant strains and its ability to form biofilm on medical associated devices. Phytochemicals acting in synergy are effective in killing the micro-organisms by lowering the doses, and synergistic compounds evade the development of resistance due to different mechanism of action. This study aims to determine the synergistic antimicrobial potential of curcumin with cinnamaldehyde, eugenol, and ellagic acid againstS. epidermidis.Curcumin with ellagic acid as well as eugenol were found to have additive antimicrobial effect, whereas, in combination, curcumin and cinnamaldehyde were found to have synergistic effect againstS. epidermidis(fractional inhibitory concentration index (FICI) = 0.5). Synergy between curcumin and cinnamaldehyde was established by time–kill kinetics and was further evaluated for antibiofilm activity. The dose required to inhibit biofilm formation was reduced to half than that needed to inhibit its planktonic culture (minimal inhibitory concentration (MIC) of curcumin = 3.12 μg/ml; MIC of cinnamaldehyde = 15.62 μg/ml; FICI = 0.248). Both curcumin and cinnamaldehyde disrupted the bacterial membrane for killing the bacteria as determined by permeability studies onEscherichia coliML-35p. [ABSTRACT FROM PUBLISHER]

Abstract: Pseudomonas aeruginosa is a gram-negative pathogen that has become an important cause of infection, especially in patients with compromised host defense mechanisms. It is frequently related to nosocomial infections such as pneumonia, urinary tract infections (UTIs) and bacteremia. The biofilm formed by the bacteria allows it to adhere to any surface, living or non-living and thus Pseudomonal infections can involve any part of the body. Further, the adaptive and genetic changes of the micro-organisms within the biofilm make them resistant to all known antimicrobial agents making the Pseudomonal infections complicated and life threatening. Pel, Psl and Alg operons present in P. aeruginosa are responsible for the biosynthesis of extracellular polysaccharide which plays an important role in cell–cell and cell–surface interactions during biofilm formation. Understanding the bacterial virulence which depends on a large number of cell-associated and extracellular factors is essential to know the potential drug targets for future studies. Current novel methods like small molecule based inhibitors, phytochemicals, bacteriophage therapy, photodynamic therapy, antimicrobial peptides, monoclonal antibodies and nanoparticles to curtail the biofilm formed by P. aeruginosa are being discussed in this review. [Copyright &y& Elsevier]

Green tea catechins and caffeine have exhibited antibacterial activity; however, their use is limited by lack of stability and effective delivery systems. Polyphenon 60 (P60) and caffeine were encapsulated in a single microemulsion (ME) formulation with an objective to lower the minimum inhibitory concentrations (MICs) of the individual agents against selected pathogens (S. epidermidis and E. coli). Combination of two natural compounds would advocate two different mechanisms on the bacterial growth thereby providing for better antibacterial activity. Thermodynamically stable ME was developed and characterized with an average particle size of 17.58 nm, further confirmed by TEM analysis. Antibacterial studies included chequerboard microdilution assay to determine the MIC and fractional inhibitory concentration (FIC) of both the natural compounds individually and in combination. MIC and FIC results indicated that the combination of the above two natural compounds was proficient in lowering the MICs of individual agents. Results of DPPH assay indicated that ME system preserved the long term antioxidative potential of P60 and caffeine. The cytotoxicity of the optimized formulation on Vero cell line by MTT assay was found to be nontoxic to mammalian cells. [ABSTRACT FROM AUTHOR]

The aim of the present study was to optimize lorazepam loaded PLGA nanoparticles (Lzp-PLGA-NPs) by investigating the effect of process variables on the response using Box-Behnken design. Effect of four independent factors, that is, polymer, surfactant, drug, and aqueous/organic ratio, was studied on two dependent responses, that is, z-average and % drug entrapment. Lzp-PLGANPs were successfully developed by nanoprecipitation method using PLGA as polymer, poloxamer as surfactant and acetone as organic phase. NPs were characterized for particle size, zeta potential, % drug entrapment, drug release behavior, TEM, and cell viability. Lzp-PLGA-NPs were characterized for drug polymer interaction using FTIR.The developed NPs showed nearly spherical shape with z-average 167-318 d-nm, PDI below 0.441, and -18.4mV zeta potential with maximum % drug entrapment of 90.1%. In vitro drug release behavior followed Korsmeyer-Peppas model and showed initial burst release of 21.7 ± 1.3% with prolonged drug release of 69.5 ± 0.8% from optimized NPs up to 24 h. In vitro drug release data was found in agreement with ex vivo permeation data through sheep nasal mucosa. In vitro cell viability study on Vero cell line confirmed the safety of optimized NPs. Optimized Lzp-PLGA-NPs were radiolabelled with Technitium-99m for scintigraphy imaging and biodistribution studies in Sprague-Dawley rats to establish nose-to-brain pathway. [ABSTRACT FROM AUTHOR]

The nucleocapsid (N) protein of Chandipura virus (CHPV) plays a crucial role in viral life cycle, besides being an important structural component of the virion through proper organization of its interactions with other viral proteins. In a recent study, the authors had mapped the associations among CHPV proteins and shown that N protein interacts with four of the viral proteins: N, phosphor protein (P), matrix protein (M), and glycoprotein (G). The present study aimed to distinguish the regions of CHPV N protein responsible for its interactions with other viral proteins. In this direction, we have generated the structure of CHPV N protein by homology modeling using SWISS-MODEL workspace and Accelrys Discovery Studio client 2.55 and mapped the domains of N protein using PiSQRD. The interactions of N protein fragments with other proteins were determined by ZDOCK rigid-body docking method and validated by yeast two-hybrid and ELISA. The study revealed a unique binding site, comprising of amino acids 1-30 at the N terminus of the nucleocapsid protein (N1) that is instrumental in its interactions with N, P, M, and G proteins. It was also observed that N2 associates with N and G proteins while N3 interacts with N, P, and M proteins. [ABSTRACT FROM AUTHOR]

Abstract: Camellia sinensis (tea) is known for its therapeutic properties (anti-inflammatory, anti-microbial, anti-tumour, anti-oxidative and anti-ageing). Although, anti-microbial properties of green tea have been studied, its role against bacterial strains related to skin infections and mechanism of action is not well understood. We focussed on exploring anti-microbial activity and the basic mechanism of aqueous green tea leaf extract on selected bacterial strains. Staphylococcus epidermidis, Micrococcus luteus, Brevibacterium linens, Pseudomonas fluorescens and Bacillus subtilis were found to be sensitive to green tea extract via disc diffusion assay (zone of inhibition ⩾7mm). Minimal inhibitory concentration (MIC) was determined via nitro blue tetrazolium (NBT) assay (0.156–0.313mg/ml). Moreover, the aqueous extract was found to be not toxic to the Vero cell-line up to a concentration of 500μg/ml. The effect of aqueous extract on adhesion of different bacteria to Vero cells indicated that it inhibits the adhesion at its MIC value. [Copyright &y& Elsevier]

Chandipura virus (CHPV) is an emerging rhabdovirus responsible for several outbreaks of fatal encephalitis among children in India. The characteristic structure of the virus is a result of extensive and specific interplay among its five encoded proteins. The revelation of interactions among CHPV proteins can help in gaining insight into viral architecture and pathogenesis. In the current study, we carried out comprehensive yeast two-hybrid (Y2H) analysis to elucidate intraviral protein-protein interactions. All of the interactions identified by Y2H were assessed for reliability by GST pull-down and ELISA. A total of eight interactions were identified among four viral proteins. Five of these interactions are being reported for the first time for CHPV. Among these, the glycoprotein (G)-nucleocapsid (N) interaction could be considered novel, as this has not been reported for any members of the family Rhabdoviridae. This study provides a framework within which the roles of the identified protein interactions can be explored further for understanding the biology of this virus at the molecular level. [ABSTRACT FROM AUTHOR]

Abstract: The four nonstructural proteins (nsPs1–4) of Chikungunya virus (CHIKV) play important roles involving enzymatic activities and specific interactions with both viral and host components, during different stages of viral pathogenesis. Elucidation of the presence and/or absence of interactions among nsPs in a systematic manner is thus of scientific interest. In the current study, each pair-wise combination among the four nonstructural proteins of CHIKV was systematically analyzed for possible interactions. Six novel protein interactions were identified for CHIKV, using systems such as yeast two-hybrid, GST pull down and ELISA, three of which have not been previously reported for the genus Alphavirus. These interactions form a network of organized associations that suggest the spatial arrangement of nonstructural proteins in the late replicase complex. The study identified novel interactions as well as concurred with previously described associations in related alphaviruses. [Copyright &y& Elsevier]

The article discusses the therapeutic properties of Solanum nigrum, a medicinal plant member of the Solanaceae family of plants. According to the author, S. nigrum is commonly known as Makoi or black nightshade and usually grows as a weed in moist habitats in different kinds of soils. The author adds that S. nigrum possesses various compounds such as glycoalkaloids, glycoproteins, and polysaccharides.

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]