Drug-resistant Pathogen Strains Research Articles

Antibacterial Efficiency of Tanacetum vulgare Essential Oil against ESKAPE Pathogens and Synergisms with Antibiotics.

Medicinal plants with multiple targets of action have become one of the most promising solutions in the fight against multidrug-resistant (MDR) bacterial infections. Tanacetum vulgare (Tansy) is one of the medicinal plants with antibacterial qualities that deserve to be studied. Thus, this research takes a closer look at tansy extract's composition and antibacterial properties, aiming to highlight its potential against clinically relevant bacterial strains. In this respect, the antibacterial test was performed against several drug-resistant pathogenic strains, and we correlated them with the main isolated compounds, demonstrating the therapeutic properties of the extract. The essential oil was extracted via hydrodistillation, and its composition was characterized via gas chromatography. The main isolated compounds known for their antibacterial effects were α-Thujone, β-Thujone, Eucalyptol, Sabinene, Chrysanthenon, Camphor, Linalool oxide acetate, cis-Carveol, trans-Carveyl acetate, and Germacrene. The evaluation of the antibacterial activity was carried out using the Kirby-Bauer and binary microdilution methods on Gram-positive and Gram-negative MDR strains belonging to the ESKAPE group (i.e., Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp.). Tansy essential oil showed MIC values ranging from 62.5 to 500 μg/mL against the tested strains. Synergistic activity with different classes of antibiotics (penicillins, cephalosporins, carbapenems, monobactams, aminoglycosides, and quinolones) has also been noted. The obtained results demonstrate that tansy essential oil represents a promising lead for developing new antimicrobials active against MDR alone or in combination with antibiotics.

Nutritional Supplements for the Control of Avian Coccidiosis – A Review

Abstract Coccidiosis is acclaimed as the most prevalent enteric parasitic ailment of poultry. It is caused by an apicomplexan protozoon of the genus Eimeria, which resides in chicken intestinal epithelium leading to intestinal damage. As a result, bloody droppings are there, feed efficiency is reduced, the growth rate is impaired, and egg production is temporarily decreased. Treatment and prevention of coccidiosis are primarily accomplished by inoculating live vaccines and administering anticoccidial drugs. Due to anticoccidials’ continuous and excessive use, the mounting issue is drug resistant Eimeria strains. The poultry industry has managed resistance-related issues by suggesting shuttle and rotation schemes. Furthermore, new drugs have also been developed and introduced, but it takes a long time and causes cost inflation in the poultry industry. Moreover, government disallows growth promoters and drugs at sub-therapeutic doses in poultry due to increased concerns about the drug residues in poultry products. These constraints have motivated scientists to work on alternative ways to control coccidiosis effectively, safely, and sustainably. Using nutritional supplements is a novel way to solve the constraints mentioned above. The intriguing aspects of using dietary supplements against coccidiosis are that they reduce the risk of drug-resistant pathogen strains, ensure healthy, nutritious poultry products, have less reliance on synthetic drugs, and are typically considered environmentally safe. Furthermore, they improve productivity, enhance nonspecific immunity, preventing oxidation of fats (acting as antioxidants) and inflammation (acting as an anti-inflammatory). The present manuscript focuses on the efficacy, possible mechanism of action, applications, and different facets of nutrition supplements (such as organic acids, minerals, vitamins, probiotics, essential oils, amino acids, dietary nucleotides, feed enzymes, and yeast derivatives) as feed additive for treating poultry coccidiosis.

Biochemistry, mechanistic intricacies, and therapeutic potential of Antimicrobial Peptides: an alternative to traditional Antibiotics.

The emergence of drug-resistant strains of pathogens becomes a major obstacle to treating human diseases. Antibiotics and antivirals are in the application for a long time but now these drugs are not much effective anymore against disease-causing drug-resistant microbes and gradually it is becoming a serious complication worldwide. The development of new antibiotics cannot be a stable solution to treat drug-resistant strains due to their evolving nature and escaping antibiotics. At this stage, antimicrobial peptides (AMPs) may provide us with novel therapeutic leads against drug-resistant pathogens. Structurally, antimicrobial peptides are mostly α-helical peptide molecules with amphiphilic properties that carry the positive charge (cationic) and it belong to host defence peptides. These positively charged AMPs can interact with negatively charged bacterial cell membranes and may cause the alteration in electrochemical potential on bacterial cell membranes and consequently lead to the death of microbial cells. In the present study, we will elaborate on the implication of AMPs in the treatment of various diseases along with their specific structural and functional properties. This review will provide information which assists in the development of new synthetic peptide analogues to natural AMPs. These analogues will eliminate the limitations of natural AMPs like toxicity and severe hemolytic activities.

Rapid, efficient auxin-inducible protein degradation in Candida pathogens.

A variety of inducible protein degradation (IPD) systems have been developed as powerful tools for protein functional characterization. IPD systems provide a convenient mechanism for rapid inactivation of almost any target protein of interest. Auxin-inducible degradation (AID) is one of the most common IPD systems and has been established in diverse eukaryotic research model organisms. Thus far, IPD tools have not been developed for use in pathogenic fungal species. Here, we demonstrate that the original AID and the second generation, AID2, systems work efficiently and rapidly in the human pathogenic yeasts, Candida albicans and Candida glabrata. We developed a collection of plasmids that support AID system use in laboratory strains of these pathogens. These systems can induce >95% degradation of target proteins within minutes. In the case of AID2, maximal degradation was achieved at low nanomolar concentrations of the synthetic auxin analog 5-adamantyl-indole-3-acetic acid. Auxin-induced target degradation successfully phenocopied gene deletions in both species. The system should be readily adaptable to other fungal species and to clinical pathogen strains. Our results define the AID system as a powerful and convenient functional genomics tool for protein characterization in fungal pathogens. IMPORTANCE Life-threatening fungal infections are an escalating human health problem, complicated by limited treatment options and the evolution of drug resistant pathogen strains. Identification of new targets for therapeutics to combat invasive fungal infections, including those caused by Candida species, is an urgent need. In this report, we establish and validate an inducible protein degradation methodology in Candida albicans and Candida glabrata that provides a new tool for protein functional characterization in these, and likely other, fungal pathogen species. We expect this tool will ultimately be useful for the identification and characterization of promising drug targets and factors involved in virulence and drug resistance.

Nanomaterials: Potential Broad Spectrum Antimicrobial Agents

Nanotechnology is a promising science with new aspects to fight and prevent various diseases using nanomaterials. The capability to expose the structure and functions of biosystems at the nanoscale level supports research leading to development in biology, biotechnology, medicine, and healthcare. This is predominantly advantageous in treating microbial infections as an alternative to antibiotics. However, widespread production, use, and misuse of antibiotics have input to the next-generation alarm for the emergence of multiple-drug-resistant pathogenic bacteria. Due to infectious diseases from these drug-resistant pathogenic strains, human mortality rates have consistently increased and are becoming an epidemic in our society. Consequently, there is a strong demand for developing novel strategies and new materials that can cope with these problems. The emergence of nanotechnology has created many new antimicrobial options. The small size of these nanomaterials is suitable for carrying out biological operations. Several metals and metal oxides, such as silver, copper, gold, zinc oxide, and iron oxide nanoparticle types have shown toxicity toward several pathogenic microbes. Metal-based nanoparticles have been broadly examined for a set of biomedical applications. According to the World Health Organization, the reduced size and selectivity of metal-based nanoparticles for bacteria have established them to be effective against pathogens causing concern. Metal-based nanoparticles are known to have nonspecific bacterial toxicity mechanisms, which not only make the development of resistance by bacteria difficult, but also widen the spectrum of antibacterial activity. Metal-based nanoparticle efficiency studies achieved so far have revealed promising results against both Gram-positive and Gram-negative bacteria. Here we discuss the potential nanomaterials to either treat microbial resistance or induce development of resistance. However, fundamental research is required to focus on the molecular mechanism causing the antimicrobial activity of nanomaterials.

Plant-Derived Natural Products as Lead Agents against Common Respiratory Diseases.

Never has the world been more challenged by respiratory diseases (RDs) than it has witnessed in the last few decades. This is evident in the plethora of acute and chronic respiratory conditions, ranging from asthma and chronic obstructive pulmonary disease (COPD) to multidrug-resistant tuberculosis, pneumonia, influenza, and more recently, the novel coronavirus (COVID-19) disease. Unfortunately, the emergence of drug-resistant strains of pathogens, drug toxicity and side effects are drawbacks to effective chemotherapeutic management of RDs; hence, our focus on natural sources because of their unique chemical diversities and novel therapeutic applications. This review provides a summary on some common RDs, their management strategies, and the prospect of plant-derived natural products in the search for new drugs against common respiratory diseases.

Rational Design of an Antimicrobial Peptide Based on Structural Insight into the Interaction of Pseudomonas aeruginosa Initiation Factor 1 with Its Cognate 30S Ribosomal Subunit.

Bacterial infections continue to represent a major worldwide health hazard following the emergence of drug-resistant pathogenic strains. Pseudomonas aeruginosa is an opportunistic pathogen causing nosocomial infections with increased morbidity and mortality. The increasing antibiotic resistance in P. aeruginosa has led to an unmet need for discovery of new antibiotic candidates. Bacterial protein synthesis is an essential metabolic process and a validated target for antibiotic development; however, the precise structural mechanism in P. aeruginosa remains unknown. In this work, the interaction of P. aeruginosa initiation factor 1 (IF1) with the 30S ribosomal subunit was studied by NMR, which enabled us to construct a structure of IF1-bound 30S complex. A short α-helix in IF1 was found to be critical for IF1 ribosomal binding and function. A peptide derived from this α-helix was tested and displayed a high ability to inhibit bacterial growth. These results provide a clue for rational design of new antimicrobials.

Antimicrobial Natural Products Produced by Soil-Derived Fungus Penicillium cremeogriseum W1-1

Ten compounds (1-10) were obtained from soil-derived Penicillium cremeogriseum W1-1 with the antimicrobial guided isolation procedure. Among them, 4 presented broad-spectrum antimicrobial activities and its preliminary mechanisms were evaluated. Compound 4 showed growth inhibition on drug-resistant pathogenic strains Escherichia coli and Candida albicans with post-contact effect (PCE), changed the morphology and membrane structure, killed cells with leakage, inhibited the growth of C. albicans by eradicating biofilms. Interestingly, the fraction containing 4 presented in vivo anti-pathogenic activities in mice, indicating this indole diterpenoid alkaloid could been used as potential antimicrobial agent.

Antimicrobial and Antioxidant Effects of Aqueous Extract of Lythrum Salicaria

Introduction: The spread of drug-resistant pathogenic strains and the increase of free radicals in food sources are major health problems worldwide. The side effects of chemical drugs have led the researchers to use natural products. In this study, the antimicrobial and antioxidant effects of leaf extract of Lythrum Salicaria have been investigated.
 Methods: This research was a basic experimental study. The aqueous extract of Lythrum Salicaria leaf was prepared by maceration method. Antimicrobial effects of the extract were evaluated by disk diffusion method. The antioxidant properties of extract were determined by DPPH and iron reducing power methods. In addition, the amounts of phenol, flavonoid and flavonol were assayed by Folin-ciocalteu and Aluminum chloride methods, respectively. Data were compared by analysis of variance (ANOVA) using SPSS version 16 software.
 Results: All of the studied strains were sensitive to the aqueous extract of Lythrum Salicaria leaf except E. coli. The mean zones of inhibition were due to the extracts on the tested bacteria obtained in ranging from 12.67 to 24.67 mm. Statistical analysis showed a significant relationship between the mean diameters of growth inhibition zone and increased extract concentrations (P < 0.001). The total phenolic content, total flavonoid content and total flavonol content values of extract were determined as 36.53 ± 1.27 mg gallic acid per gram extract, 5.2 ± 0.4 mg quercetin per gram extract and 20.8 ± 0.95 quercetin per gram extract, respectively.
 Conclusion: Extract of Lythrum Salicaria can be a good alternative to common antibiotics resistant. The acceptable antioxidant effects of the Lythrum Salicaria extract can be introduced it as an inhibitor of diabetes and cancer.

Bacteriocin isolated from the natural inhabitant of Allium cepa against Staphylococcus aureus.

Extensive usage of antibiotics has led to the emergence of drug-resistant strains of pathogens and hence, there is an urgent need for alternative antimicrobial agents. Antimicrobial Peptides (AMPs) of bacterial origin have shown the potential to replace some conventional antibiotics. In the present study, an AMP was isolated from Bacillus subtilis subsp. spizizenii strain Ba49 present on the Allium cepa, the common onion and named as peptide-Ba49. The isolated AMP was purified and characterized. The purified peptide-Ba49, having a molecular weight of ~ 3.3kDa as determined using mass spectroscopy, was stable up to 121°Cand in the pH range of 5-10. Its interaction with protein degrading enzymes confirmed the peptide nature of the molecule. The peptide exhibited low minimum inhibitory concentration (MIC) against Staphylococcus aureus and its (Methicillin-resistant Staphylococcus aureus) MRSA strains (MIC, 2-16µM/mL). Further, time kill kinetic assay was performed and analysis of the results of membrane depolarization and permeabilization assays (TEM, DiBAC4 (3) and PI) suggested peptide-Ba49 to be acting through the change in membrane potential leading to disruption of S. aureus membrane. Additionally, cytotoxicity studies of peptide-Ba49, carried out using three mammalian cell lines viz. HEK 293T, RAW 264.7, and L929, showed limited cytotoxicity on these cell lines at a concentration much higher than its MIC values. All these studies suggested that the AMP isolated from strain Ba49 (peptide-Ba49) has the potential to be an alternative to antibiotics in terms of eradicating the pathogenic as well as drug-resistant microorganisms.

Glycol and Phosphate Depot Forms of 4- and/or 5-Modified Nucleosides Exhibiting Antibacterial Activity

Resistance developed to the majority of drugs used to treat infectious diseases warrants the design of new compounds effective against drug-resistant strains of pathogens. Recently, several groups of modified nucleosides have been synthesized and showed significant antibacterial activity in vitro, but their further studies were difficult to undertake because of their low solubility in aqueous solutions. Nevertheless, new compounds, well soluble in water-organic solutions, were synthesized and found to be more effective in inhibiting the growth of Gram-positive bacteria and mycobacteria. The water-soluble forms of modified nucleosides under study were assumed to be their depot forms. To check the assumption, the compounds were tested for hydrolysis in various media and their molecular docking was performed into the active center of the putative target, Mycobacterium tuberculosis flavin-dependent thymidylate synthase ThyX. Computer modelling showed that the water-soluble analogs do not act as ThyX inhibitors, supporting the assumption of their depot nature. The compounds were resistant to chemical hydrolysis but were hydrolyzed when incubated with porcine liver carboxylesterase, human serum, or Staphylococcus aureus 209P. The results demonstrate that the compounds are most likely depot forms of modified nucleosides.

Synthesis of zinc oxide and silver nanoparticles using ficus palmata - Forssk leaf extracts and assessment of antibacterial activity

Multidrug resistance of bacteria is an emerging human health hazard and warrants development of novel antibacterial agents with more effective mode of action. Here, zinc oxide and silver nanomaterials were prepared using Ficus palmata Forssk leaf extract with efficient antibacterial activity. SEM coupled with EDS confirmed the spherical symmetry with average particle diameter 50 to 65 nm while the XRD confirmed crystalline face centered cubic structure of silver and hexagonal crystallize phase of zinc oxide nanoparticles. Antibacterial activity was evaluated for 8 pathogenic bacterial strains including 3 drug resistant pathogenic strains. The nanoparticles showed enhanced growth inhibition for resistant strains in comparison with the broad-spectrum antibiotics i.e. roxithromycin and cefixime. Minimum inhibitory concentration in μg.mL<sup>-1</sup> of silver nanoparticles was found to be as low as 33.3 for resistant Streptococcus haemolyticus; 11.1 for Staphylococcus aureus and E Coli; and 3.7 μg.mL<sup>-1</sup> for resistant Pseudomonas aeruginosa. Similarly, the minimum inhibitory concentration of zinc oxide nanoparticles was found to be 100 μg.mL<sup>-1</sup> against resistant Streptococcus haemolyticus and Staphylococcus aureus; 11.1 μg.mL<sup>-1</sup> for resistant Pseudomonas aeruginosa; and 3.7 μg.mL<sup>-1</sup> against resistant E coli. Ficus palmata Forssk leaf extracts can be explored effectively for synthesizing active antibacterial nanomaterials as a non-toxic and environmentally benign synthesis route.

Systematic comparison of coexistence in models of drug-sensitive and drug-resistant pathogen strains

A number of mathematical models have recently been proposed to explain empirical trends of pathogen diversity. In particular, long-term coexistence of both drug-sensitive and drug-resistant variants of a single pathogen is something of a mystery, given that simple models of pathogens competing for the same ecological niche predict competitive exclusion, and more complex models admitting coexistence require assumptions that may not be justified. Coinfection is among the candidate mechanisms to generate coexistence, as it occurs in many pathogens and provides the opportunity for strains to interact directly. Recently, coinfection and competitive release have been described as creating a form of negative frequency-dependent selection that promotes coexistence, and a range of models containing coinfection have been proposed as having generic stable coexistence of multiple strains. This abundance of new models presents the challenge of comparison and interpretation. To this end, we describe a dimensionless quantity that can be used to compare the amount of coexistence generated by different models. We focus on models that include coinfection, although this framework could be generalized to a larger class of structured models.

Disease control efficacy of 32,33-didehydroroflamycoin produced by Streptomyces rectiviolaceus strain DY46 against gray mold of tomato fruit

Despite the efficacy of synthetic fungicides in controlling postharvest diseases, public concerns regarding chemical residues in food and an increase in drug-resistant strains of pathogens have led to a need for new agents to control postharvest diseases. The current study was performed to find control agents of microbial origin that are effective on gray mold of tomato fruits. We recently isolated Streptomyces rectiviolaceus DY46, which has antagonistic activity against various plant pathogenic fungi. The incidence of gray mold of tomato fruits was markedly reduced by 80.0% in tomatoes treated with the cell extract of Streptomyces rectiviolaceus DY46 compared with the control tomatoes. The active ingredient was purified from the cell extract of DY46 and identified to be 32,33-didehydroroflamycoin (DDHR). DDHR displayed MICs (minimal inhibitory concentrations) against the mycelial growth of various plant pathogenic fungi at concentrations of 8–64 mg L−1. The incidence of gray mold in tomato fruits inoculated with conidial suspension (104 conidia mL−1) of Botrytis cinerea was markedly reduced by 88.9% in tomatoes treated with DDHR (100 mg L−1) compared with the control. The DDHR residue in tomato fruit was significantly diminished 2 d after treatment. These results show that DDHR would be relatively safe for use as a postharvest fungicide.

Induction of autophagy through CLEC4E in combination with TLR4: an innovative strategy to restrict the survival of Mycobacterium tuberculosis

ABSTRACT Host-directed therapies are gaining considerable impetus because of the emergence of drug-resistant strains of pathogens due to antibiotic therapy. Therefore, there is an urgent need to exploit alternative and novel strategies directed at host molecules to successfully restrict infections. The C-type lectin receptor CLEC4E and Toll-like receptor TLR4 expressed by host cells are among the first line of defense in encountering pathogens. Therefore, we exploited signaling of macrophages through CLEC4E in association with TLR4 agonists (C4.T4) to control the growth of Mycobacterium tuberculosis (Mtb). We observed significant improvement in host immunity and reduced bacterial load in the lungs of Mtb-infected mice and guinea pigs treated with C4.T4 agonists. Further, intracellular killing of Mtb was achieved with a 10-fold lower dose of isoniazid or rifampicin in conjunction with C4.T4 than the drugs alone. C4.T4 activated MYD88, PtdIns3K, STAT1 and RELA/NFKB, increased lysosome biogenesis, decreased Il10 and Il4 gene expression and enhanced macroautophagy/autophagy. Macrophages from autophagy-deficient (atg5 knockout or Becn1 knockdown) mice showed elevated survival of Mtb. The present findings also unveiled the novel role of CLEC4E in inducing autophagy through MYD88, which is required for control of Mtb growth. This study suggests a unique immunotherapeutic approach involving CLEC4E in conjunction with TLR4 to restrict the survival of Mtb through autophagy. Abbreviations 3MA: 3 methyladenine; AO: acridine orange; Atg5: autophagy related 5; AVOs: acidic vesicular organelles; BECN1: beclin 1, autophagy related; BMDMs: bone marrow derived macrophages; bw: body weight; C4.T4: agonists of CLEC4E (C4/TDB) and TLR4 (T4/ultra-pure-LPS); CFU: colony forming unit; CLEC4E/Mincle: C-type lectin domain family 4, member e; CLR: c-type lectin receptor; INH: isoniazid; LAMP1: lysosomal-associated membrane protein 1; MφC4.T4: Mtb-infected C4.T4 stimulated macrophages; MAP1LC3/LC3: microtubule-associated protein 1 light chain 3; MDC: monodansylcadaverine; MTOR: mechanistic target of rapamycin kinase; MYD88: myeloid differentiation primary response 88; NFKB: nuclear factor of kappa light polypeptide gene enhance in B cells; NLR: NOD (nucleotide-binding oligomerization domain)-like receptors; PFA: paraformaldehyde; PPD: purified protein derivative; PtdIns3K: class III phosphatidylinositol 3-kinase; RELA: v-rel reticuloendotheliosis viral oncogene homolog A (avian); RIF: rifampicin; RLR: retinoic acid-inducible gene-I-like receptors; TDB: trehalose-6,6´-dibehenate; TLR4: toll-like receptor 4; Ultra-pure-LPS: ultra-pure lipopolysaccharide-EK; V-ATPase: vacuolar-type H+ ATPase.

Efficacy of Andrographis paniculata against AmpC producing multi drug resistant E. coli

Abstract Andrographis paniculata (Acanthaceae) is a well-known medicinal plant and is conventionally used in herbal medicine for the treatment of several diseases such as cancer, metabolic disorders, skin infections, respiratory tract infections, etc. In this present study, ethyl acetate extract of A. paniculata was used to test its efficacy on controlling growth and biofilm formation in multi drug resistant pathogenic strains of Escherichia coli through micro-dilution method. Antibiotic susceptibility screening through disc diffusion revealed that these E. coli strains were resistant towards 13 out of 15 antibiotics used. It was found that 250 and 500 μg/ml of crude ethyl acetate (CEA) extract effectively reduced the growth and biofilm formation as compared to 62 and 125 μg/ml. The strains treated with CEA extract of A. paniculata reduced extracellular polysaccharide (EPS) production and downregulated the expression of AmpC gene. The results suggested that CEA extract of A. paniculata can be explored to utilize as an alternative to antibiotics.

ImmtorLig_DB: repertoire of virtually screened small molecules against immune receptors to bolster host immunity

Host directed therapies to boost immunity against infection are gaining considerable impetus following the observation that use of antibiotics has become a continuous source for the emergence of drug resistant strains of pathogens. Receptors expressed by the cells of immune system play a cardinal role in initiating sequence of events necessary to ameliorate many morbid conditions. Although, ligands for the immune receptors are available; but their use is limited due to complex structure, synthesis and cost-effectiveness. Virtual screening (VS) is an integral part of chemoinformatics and computer-aided drug design (CADD) and aims to streamline the process of drug discovery. ImmtorLig_DB is a repertoire of 5000 novel small molecules, screened from ZINC database and ranked using structure based virtual screening (SBVS) against 25 immune receptors which play a pivotal role in defending and initiating the activation of immune system. Consequently, in the current study, small molecules were screened by docking on the essential domains present on the receptors expressed by cells of immune system. The screened molecules exhibited efficacious binding to immune receptors, and indicated a possibility of discovering novel small molecules. Other features of ImmtorLig_DB include information about availability, clustering analysis, and estimation of absorption, distribution, metabolism, and excretion (ADME) properties of the screened small molecules. Structural comparisons indicate that predicted small molecules may be considered novel. Further, this repertoire is available via a searchable graphical user interface (GUI) through http://bioinfo.imtech.res.in/bvs/immtor/.

Identification of potential drug targets and vaccine candidates in Clostridium botulinum using subtractive genomics approach.

A subtractive genomic approach has been utilized for the identification of potential drug targets and vaccine candidates in Clostridium botulinum, the causative agent of flaccid paralysis in humans. The emergence of drug-resistant pathogenic strains has become a significant global public health threat. Treatment with antitoxin can target the neurotoxin at the extracellular level, however, can't converse the paralysis caused by botulism. Therefore, identification of drug targets and vaccine candidates in C. botulinum would be crucial to overcome drug resistance to existing antibiotic therapy. A total of 1729 crucial proteins, including chokepoint, virulence, plasmid and resistance proteins were mined and used for subtractive channel of analysis. This analysis disclosed 15 potential targets, which were non-similar to human, gut micro flora, and anti-targets in the host. The cellular localization of 6 targets was observed in the cytoplasm and might be used as a drug target, whereas 9 targets were localized in extracellular and membrane bound proteins and can be used as vaccine candidates. Furthermore, 4 targets were observed to be homologous to more than 75 pathogens and hence are considered as broad-spectrum antibiotic targets. The identified drug and vaccine targets in this study would be useful in the design and discovery of novel therapeutic compounds against botulism.

Prioritizing and modelling of putative drug target proteins of Candida albicans by systems biology approach.

Candida albicans (Candida albicans) is one of the major sources of nosocomial infections in humans which may prove fatal in 30% of cases. The hospital acquired infection is very difficult to treat affectively due to the presence of drug resistant pathogenic strains, therefore there is a need to find alternative drug targets to cure this infection. In silico and computational level frame work was used to prioritize and establish antifungal drug targets of Candida albicans. The identification of putative drug targets was based on acquiring 5090 completely annotated genes of Candida albicans from available databases which were categorized into essential and non-essential genes. The result indicated that 9% of proteins were essential and could become potential candidates for intervention which might result in pathogen eradication. We studied cluster of orthologs and the subtractive genomic analysis of these essential proteins against human genome was made as a reference to minimize the side effects. It was seen that 14% of Candida albicans proteins were evolutionary related to the human proteins while 86% are non-human homologs. In the next step of compatible drug target selections, the non-human homologs were sequentially compared to the human microbiome data to minimize the potential effects against gut flora which accumulated to 38% of the essential genome. The sub-cellular localization of these candidate proteins in fungal cellular systems indicated that 80% of them are cytoplasmic, 10% are mitochondrial and the remaining 10% are associated with the cell wall. The role of these non-human and non-gut flora putative target proteins in Candida albicans biological pathways was studied. Due to their integrated and critical role in Candida albicans replication cycle, four proteins were selected for molecular modeling. For drug designing and development, four high quality and reliable protein models with more than 70% sequence identity were constructed. These proteins are used for the docking studies of the known and new ligands (unpublished data). Our study will be an effective framework for drug target identifications of pathogenic microbial strains and development of new therapies against the infections they cause.

A study on antibacterial effect of grape seed extracts in common clinical and drug resistant isolates

<p class="abstract"><strong>Background:</strong> Grape seeds are proposed to have antimicrobial activity, antioxidant effect and various other benefits to mankind. A study was done to assess the antibacterial effect of grape seed extract against common clinical isolates and drug resistant pathogenic strains.</p><p class="abstract"><strong>Methods:</strong> Grape seed extract prepared was investigated for its antibacterial effect against 65 bacterial isolates obtained from clinical specimens by agar well diffusion assay and the results were compared with routinely used antibiotics namely, Gentamicin for the common clinical isolates, Vancomycin for MRSA strains and Amikacin for ESBL organisms respectively.</p><p class="abstract"><strong>Results:</strong> Grape seed extract produced moderate zone of inhibition ranging between 11-15 mm among the 35 test common clinical isolates namely <em>Staphylococcus aureus, Escherichia coli, Klebsiella </em>sp<em> </em>and<em> Pseudomonas aeruginosa</em>. <em>E.coli </em>showed the highest susceptibility with zone ranging from 12-14 mm with increasing concentration of the extract starting from2 mg/ml to the highest being 20 mg/ml. Among the 30 drug resistant pathogenic strains like MRSA and ESBL producing organisms, the grape seed extract was found to be effective against 3 out of the 10 of MRSA and 2 out of the 10 of ESBL-<em>E.coli</em> at the highest concentration of 20 mg/ml. However, ESBL producing <em>Klebsiella </em>species were found to be resistant even to the highest concentration of the extract. </p><p><strong>Conclusions:</strong> The results provide evidence that the grape seed extract could be a potential antibacterial agent and this effect can further be made evident with improved methodologies.</p>