Comprehensive quality assessment of peppermint oils and commercial products: An integrated approach involving conventional and chiral GC/MS coupled with chemometrics.

Peppermint essential oil (EO) has a multitude of applications, such as a fragrance in cosmetics, personal care and industrial products, or as a flavoring ingredient in food and beverages. Despite its popularity and economic significance, peppermint EO is often adulterated to reduce production costs and to increase profits. Although the ISO standard for peppermint EO exists, detecting sophisticated forms of adulteration remains challenging.The current study used conventional and chiral GC/MS analysis of volatiles compounds, and chemometric techniques to evaluate an extensive set of authentic peppermint EO (n = 22) and commercial products (n = 36) purported to contain peppermint EO. Specifically, thirty-six terpenoids were examined in each sample and compared with the ISO standard. Fifty-three percent of the selected commercial products did not meet the ISO specifications and the ratio between menthone/isomenthone was proven to be a good indicator for authentication and adulteration detection. Chiral GC/MS was further employed to measure eight terpenoids: α-pinene, ß-pinene, limonene, menthol, menthone, isomenthone, pulegone, and menthyl acetate. The enantiomeric compositions of 27 commercial products were above or below the norm measured from authentic peppermint EOs. Of the 27 samples, eight met the ISO standard. A sample class prediction (SCP) model based on partial least squares-discriminant analysis (PLS-DA) of conventional GC/MS data was constructed using authentic peppermint EOs and cornmint EOs. The model can distinguish the most common types of peppermint EOs (US, India, and US/India blend) and cornmint EOs sold in the US market. After construction, the SCP model was then used to analyze commercial samples. One sample, which passed both ISO specification and chiral analysis, was identified as outlier by the SCP model. Overall, the applicability of combining both conventional and chiral GC/MS along with chemometric tools has been successfully demonstrated to address the overall quality of peppermint EOs in commerce and may help combat sophisticated adulteration.

Discovery, Synthesis, and Optimization of 1,2,4-Triazolyl Pyridines Targeting Mycobacterium tuberculosis.

The rise in multidrug resistant tuberculosis cases underscores the urgent need to develop new treatment strategies for tuberculosis. Herein, we report the discovery and synthesis of a new series of compounds containing a 3-thio-1,2,4-triazole moiety that show inhibition of Mycobacterium tuberculosis (Mtb) growth and survival. Structure-activity relationship studies led us to identify several potent analogs displaying low micromolar to nanomolar inhibitory activity, specifically against Mtb. The potent analogs demonstrated no cytotoxicity in mammalian cells at over 100 times the effective concentration required in Mtb and were bactericidal against Mtb during infection of macrophages. In the exploratory ADME investigations, we observed suboptimal ADME characteristics, which prompted us to identify potential metabolic liabilities for further optimization. Our preliminary investigations into the mechanism of action suggest that this series is not engaging the promiscuous targets that arise from many phenotypic screens. We selected for resistant mutants with the nanomolar potent nitro-containing compound 20 and identified resistant isolates with mutations in genes required for coenzyme F420 biosynthesis and the nitroreductase Ddn. This suggests that the aromatic nitro-1,2,4-triazolyl pyridines are activated by F420-dependent Ddn activity, similar to the nitro-containing TB drug pretomanid. We were able to circumvent the requirement for F420-dependent Ddn activity using compounds that contained non-nitro groups, identifying a key feature to be modified to avoid this predominant resistance mechanism. These studies provide the foundation for the development of a new class of 1,2,4-triazole compounds for the treatment of tuberculosis.

Identification of potential non-nucleoside MraY inhibitors for tuberculosis chemotherapy using structure-based virtual screening.

The efforts to limit the spread of the tuberculosis epidemic have been challenged by the rise of drug-resistant strains of Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis. It is critical to discover new chemical scaffolds acting on novel or unexploited targets to beat this drug-resistant pathogen. MraY (phospho-MurNAc-pentapeptide translocase or translocase I) is an in vivo validated target for antibacterials-discovery. MraY is inhibited by nucleoside-based natural products that suffer from poor in vivo efficacy. The current study is focused on discovering novel chemical entities, particularly, non-nucleoside small molecules, as MraYMtb inhibitors possessing antituberculosis activity. In the absence of any reported X-ray crystal structures of MraYMtb, we used a homology model-based virtual screening approach combined with the ligand-based e-pharmacophore screening. We screened ∼12 million commercially available compounds from the ZINC15 database using GOLD software. The resulting hits were filtered using a 2-pronged screening method comprising e-pharmacophore hypotheses and docking against the MraYMtb homology model using Glide. Further clustering based on Glide scores and optimal binding interactions resulted in 15 in silico hits. We performed molecular dynamics (MD) simulations for the three best-ranking compounds and one other poorer-ranking compound, out of the 15 in silico hits, to analyze the interaction modes in detail. The MD simulations indicated stable interactions between the compounds and key residues in the MraY active site that are crucial for maintaining the enzymatic activity. These in silico hits could advance the antibacterial drug discovery campaign to find new MraY inhibitors for tuberculosis treatment.Communicated by Ramaswamy H. Sarma.

Studies Directed toward the Stereoselective Synthesis of Cytospolide E.

Our exhaustive effort toward the total synthesis of cytotoxic marine nonanolide cytospolide E has been detailed. To achieve this synthesis, we have explored both the ring-closing metathesis and lactonization-based macrocyclization strategies using a variety of precursors. Unfortunately, none of them provided the desired product. The ring-closing metathesis approach provided mainly the macrocycle with Z-olefin, whereas the macrolactonization strategy culminated in 8-epi-9-epi-cytospolide E following the regioselective formation of a 10-membered macrocycle over a 9-membered macrocycle.

Total Synthesis of Cytospolide Q.

A flexible and convergent strategy for the stereoselective total synthesis of bioactive marine natural product cytospolide Q has been developed. The key features of this synthesis include Evans anti-aldol reaction for the installation of C-2 and C-3 stereocenters and cycloetherification via epoxide opening followed by concomitant lactonization for the construction of tetrahydrofuran and γ-butyrolactone scaffolds. This synthetic study also revealed that protected oxygenated functionality (methyl ester or benzyl ether) at C-1 position participated readily in epoxide opening.

Total Synthesis of Reported Structure of Baulamycin A and Its Congeners.

A convergent and flexible strategy for the stereoselective total synthesis of the reported structure of baulamycin A and its congeners has been developed for the first time. Synthetic highlights include a Crimmins aldol reaction to construct the C-1' and C-14 centers, a Crimmins acetate aldol reaction to generate the hydroxy group at the C-13 position, Horner-Wadsworth-Emmons olefination to form the C9-C10 bond, and Evans methylation to install the C-8 center. This synthetic study disclosed that the reported structure of baulamycin A needs to be revised, as its spectroscopic data are not identical with those of the synthetic baulamycin A.

Production and estimation of alkaline protease by immobilized Bacillus licheniformis isolated from poultry farm soil of 24 Parganas and its reusability.

Microbial alkaline protease has become an important industrial and commercial biotech product in the recent years and exerts major applications in food, textile, detergent, and pharmaceutical industries. By immobilization of microbes in different entrapment matrices, the enzyme produced can be more stable, pure, continuous, and can be reused which in turn modulates the enzyme production in an economical manner. There have been reports in support of calcium alginate and corn cab as excellent matrices for immobilization of Bacillus subtilis and Bacillus licheniformis, respectively. This study has been carried out using calcium alginate, κ-carrageenan, agar-agar, polyacrylamide gel, and gelatin which emphasizes not only on enzyme activity of immobilized whole cells by different entrapment matrices but also on their efficiency with respect to their reusability as first attempt. Gelatin was found to be the best matrix among all with highest enzyme activity (517 U/ml) at 24 h incubation point and also showed efficiency when reused.

Stereoselective total synthesis of cytospolide P.

A short and convergent stereoselective total synthesis of biologically potent cytospolide P has been accomplished from acrolein. The salient features of our synthetic strategy include modified Crimmins aldols, Yamaguchi esterification, and Grubbs ring-closing metathesis reaction.

Progesterone receptor agonists and antagonists as anticancer agents.

Progesterone is a major female steroid hormone produced by the ovarian corpus luteum and by the placental syncytiotrophoblast during the second trimester. The biological effects of this steroid hormone are mediated by the ubiquitously expressed progesterone receptor. The exact link between progesterone and female reproductive organ cancer is a controversial issue with various cross-talks. The present review summarizes recent trends in the development of some (anti)progestagen in the cure and management of breast and uterine cancers.

Anti-androgenic endocrine disrupting activities of chlorpyrifos and piperophos.

The present work describes the screening and characterization of some common endocrine disrupting chemicals for their (anti)androgenic activities. Various chemicals (mostly pesticides and pharmaceuticals) were screened with the NIH3T3 cell line stably expressing human androgen receptor (hAR) and luciferase reporter gene for their ability to stimulate luciferase activity or inhibit the response that was evoked by 0.4nM testosterone. The most potent anti-androgenic compounds identified in our assay included chlorpyrifos, endosulfan and piperophos. Finally, the chemicals were analyzed for their effects on steriodogenesis in rat Leydig cells. Piperophos and chlorpyrifos showed a significant decrease in testosterone biosynthesis by Leydig cells. RT-PCR studies showed decrease in the expression of key steroidogenic enzymes: cytochrome P450scc, 3beta-HSD and 17beta-HSD and immunoblot analysis demonstrated a decrease in steroidogenic acute regulatory (StAR) protein expression by both these chemicals. Chlorpyrifos also showed a decrease in LH receptor stimulated cAMP production. In conclusion, we demonstrate that commonly used pesticides like chlorpyrifos and piperophos pose serious threat to male reproductive system by interfering at various levels of androgen biosynthesis.

Biodegradation of pyridine by the new bacterial isolates S. putrefaciens and B. sphaericus.

In this study, two bacterial strains capable of utilizing pyridine as a sole carbon source were isolated from biofilters. Based on the biochemical test, the organisms were identified as Shewanella putrefaciens and Bacillus sphaericus. In liquid cultures, S. putrefaciens and B. sphaericus degraded pyridine quite effectively up to 500 mg L(-1). S. putrefaciens degrades 500 mg L(-1) of pyridine completely within 140 h, whereas the B. sphaericus degrades 500 mg L(-1) of pyridine only nearly 75% and takes a longer duration of 150 h. S. putrefaciens used pyridine as sole carbon and energy source better than B. sphaericus. Monod's and Haldane's inhibitory growth models were used to obtain maximum specific growth rate (micro(max)), half saturation (K(s)) and substrate inhibition (K(i)) constant for pyridine by using S. putrefaciens and B. sphaericus. The high value of K(i) for S. putrefaciens than B. sphaericus indicates that the inhibition effect can be observed only in a high concentration range. The S. putrefaciens degrades pyridine with a faster rate than B. sphaericus. S. putrefaciens can be used effectively for the treatment of pyridine bearing wastewater and as an inoculum in a biofilter treating pyridine-laden gas.

Screening of some anti-progestin endocrine disruptors using a recombinant yeast based in vitro bioassay.

The present study was aimed to develop a sensitive, fast and user friendly progesterone receptor transactivation assay using recombinant yeast cells, Saccharomyces cerevisiae, modified to express human progesterone receptor (PR) and progesterone response element (PRE) driving the expression of green fluorescent protein. Stimulation of cells with increasing concentrations of progesterone resulted in significant elevation in fluorescence activity, with the minimum effective dose of progesterone being 0.1 nM. RU486, significantly inhibited progesterone induced transactivation and non-progesterogenic steroids failed to transactivate PR till 10 microM concentrations. About 7 different chemicals (mostly pesticides or their metabolites) like DDT and its metabolites, nonylphenol, endosulfan were screened in this assay system for their role in transactivation and they were all found to be anti-progestative and IC50 values within the range of 3-20 microM. Further, the assay was used to analyze the endocrine disrupting activity of extracted water samples from leather industries known for their high content of various chemicals and it was found to be rich in anti-progestative compounds. It resulted in about 30% reduction in transactivation. In conclusion, we demonstrated that this yeast based bioassay provides a rapid and robust assay for high throughput screening of (anti)progestative compounds from various sources.

Combined removal of BTEX in air stream by using mixture of sugar cane bagasse, compost and GAC as biofilter media.

Biofiltration of air stream containing mixture of benzene, toluene, ethyl benzene and o-xylene (BTEX) has been studied in a lab-scale biofilter packed with a mixture of compost, sugar cane bagasse and granulated activated carbon (GAC) in the ratio 55:30:15 by weight. Microbial acclimation was achieved in 30 days by exposing the system to average BTEX inlet concentration of 0.4194 gm(-3) at an empty bed residence time (EBRT) of 2.3 min. Biofilter achieved maximum removal efficiency more than 99% of all four compounds for throughout its operation at an EBRT of 2.3 min for an inlet concentration of 0.681 gm(-3), which is quite significance than the values reported in the literature. The results indicate that when the influent BTEX loadings were less than 68 gm(-3)h(-1) in the biofilter, nearly 100% removal could be achieved. A maximum elimination capacity (EC) of 83.65 gm(-3)h(-1) of the biofilter was obtained at inlet BTEX load of 126.5 gm(-3)h(-1) in phase IV. Elimination capacities of BTEX increased with the increase in influent VOC loading, but an opposite trend was observed for the removal efficiency. The production of CO(2) in each phase (gm(-3)h(-1)) was also observed at steady state (i.e. at maximum removal efficiency). Moreover, the high concentrations of nitrogen in the nutrient solution may adversely affect the microbial activity possibly due to the presence of high salt concentrations. Furthermore, an attempt was also made to isolate the most profusely grown BTEX-degrading strain. A Gram-positive strain had a high BTEX-degrading activity and was identified as Bacillus sphaericus by taxonomical analysis, biochemical tests and 16S rDNA gene analysis methods.

Development of a yeast-based assay to determine the (anti)androgenic contaminants from pulp and paper mill effluents in India.

We have constructed an efficient and reliable yeast-based detection system to evaluate the androgenic activity of endocrine disruptors from pulp and paper mill effluents (PPME). This system consists of human androgen receptor and androgen response elements driven ß-galactosidase genes transformed in yeast, Saccharomyces cerevisiae. The transcriptional activation by known androgens, correlated with androgenic activities as measured by other assay systems. This assay system when applied to evaluate anti-androgenic activities, the known anti-androgens effectively inhibited reporter gene induction by testosterone. The specificity of the assay was tested by incubating the transformed cells with supraphysiological concentrations of non-androgenic steroids and none of them gave a significant response. The extracted PPME from five different mills demonstrated strong androgenic activities (about five- to eight-folds over control). These results suggest that PPME are rich in androgenic chemicals and the employed detection system could be applicable to primary screening for effectors on androgen receptor functions.

Higher prevalence of OCA1 in an ethnic group of eastern India is due to a founder mutation in the tyrosinase gene.

PURPOSE: Oculocutaneous albinism (OCA) is a group of autosomal recessive disorders characterized by deficient synthesis of melanin pigment and associated with common developmental abnormalities of the eye. It is one of the major causes of childhood blindness in India. The disease is common among an ethnic group (Tili) of Eastern India, which represents about 12.56% of the Bankura district population (approximately 0.4 million) of West Bengal. The purpose of the study was to investigate the molecular lesions causing OCA within this ethnic group for the unequivocal diagnosis of the carriers and attempt to decipher the cause for the high prevalence of OCA. METHODS: Fourteen OCA-affected Tili families consisting a total of 161 individuals, including 26 patients, were recruited for the study. A lack of tyrosinase (TYR) activity among all the patients was ascertained by the tyrosinase hair bulb assay. Mutation screening in the tyrosinase gene (TYR) was done by single strand conformational polymorphism (SSCP) and DNA sequencing. The restriction fragment length polymorphism (RFLP) assay was carried out to determine the frequency of the pathogenic changes among the normal individuals. Haplotype analysis was performed at the TYR locus using a set of informative microsatellite and SNP markers. RESULTS: All the patients were homozygous for a null mutation (c.832C>T, Arg278stop) in TYR exon 2, which might cause a complete loss of enzyme activity. The mutation occurred in the same haplotype background. The frequency of the disease in this ethnic group was estimated to be significantly higher than the world average. CONCLUSIONS: OCA1 in the Tili population is due to the occurrence of a founder mutation in the TYR as indicated by haplotype analysis. Higher prevalence of the mutation in the population group is due to marriage within the same community. The diagnostic RFLP assay can be utilized for genetic counseling and thereby will help to reduce the disease load on the population.