PURIFICATION AND CYTOTOXICITY STUDY OF LOVASTATIN FROM SOIL FUNGI

  Objective: The objective of the present study is to evaluate the anticancer potential of lovastatin obtained from fungal source.Methods: About 15 fungal cultures were isolated from soil samples collected from Bharathiar University, India, and all are identified and characterized through microscopic characterization. Lovastatin producing capability was confirmed through bioassay against Saccharomyces cerevisiae, and the ability of selected fungus to produce lovastatin was further confirmed by high-performance liquid chromatography. Maximum lovastatin producing fungi were further selected for purification (overloaded elution chromatography) and characterization done using inhibition rate (IR). 5-diphenyl tetrazolium bromide (MTT) assay using A549 cell line was performed for antitumor activity evaluation.Results: Among the 15 isolates, Aspergillus flavus exhibited the maximum zone of inhibition (1.5 cm) against the test organism through solid-state fermentation. The resemblance in retention time (RT) of peak shown in chromatograms of standard lovastatin (RT=25.1 minutes) and sample (RT=25.1 minutes) were similar. This confirmed the presence of lovastatin in the selected fungal isolate (A. flavus). The presence of two functional groups in lovastatin C=O and O-H was confirmed by IR spectrum 50% of cell lysis was observed in MTT assay.Conclusion: Lovastatin obtained from soil fungi is capable of producing lovastatin in good proportions. Obtained fungal lovastatin exhibited significant antitumor activity against A549 cell line. Like other biological derivatives, lovastatin from soil fungi had greater potential in anticancer activity, and further biosynthetic pathway insights in their production can improve the yield which aid in large scale production

Histomorphological perspectives of preputial and clitoral glands of soft-furred field rat Millardia meltada

The present study was an attempt to understand the sexual dimorphism of the integumentary scent glands of soft-furred fi eld rat Millardia meltada from the perspectives of anatomy, morphology and histology with view to correlate with the sex-specifi c pheromones they produce. The scent gland of male is known as preputial gland, and female, the clitoral gland. The rats, that are agricultural pests were fi eld caught, the glands of males and females of almost identical size were dissected out, and subjected to gravimetric, morphometric and histological analyses. Both glands are yellowish-brown, pear-shaped, and dorsoventrally compressed. The mean weight, length and width of preputial glands are signifi cantly (p < 0.05) larger than that of the clitoral glands. The preputial gland is composed of sebaceous glandular lobules and apocrine glandular lobules whereas the clitoral gland is formed only of sebaceous glandular lobules. The sebaceous glandular lobules of both preputial and clitoral glands are fi lled with a wax-like material. Thus, the scent glands of the soft-furred male fi eld rats exhibit sexual dimorphism in respect histoarchitecture of the glands and the nature of the secretory material. This sexual dimorphism of the scent glands may refl ect control by male and female sex hormones impinging on specifi c roles as sex attractant pheromones

A decline in avian cytokine expression with age revealed by commercially available cytokine array

Cytokines are secreted immunomodulators that are key regulators of the avian immune response. Currently, the most commonly used method to follow cytokine expression is qPCR, which measures cellular levels of mRNA, rather their extracellular circulating levels. Here we present a commercially available cytokine array designed to assay circulating expression levels of multiple cytokines and immunomodulators simultaneously. Upon minor modifications to the manufacturer protocol, background noise was reduced, leading to a significant increase in the sensitivity of the device. Our data indicate that the array is reliable and produce consistent data between biological repeats. We tested the reproducibility of the array in a biologically relevant context by assessing age-related changes in circulating cytokines. While individual features did not show a consistent pattern, our data revealed a consistent decline in the median of all cytokine values, supporting the validity of the array in studying biological processes

Exploration of salivary proteins in buffalo : an approach to find marker proteins for estrus

Saliva is considered as the best source of biological material for biomarker discovery studies since it is noninvasive in comparison to other body sources. Usually buffalo cannot precisely express estrus signals. Hence, there is a need for concise methods to detect the time of estrus to ensure the success of artificial insemination. Therefore, we have established a reference proteome map on the whole saliva of buffalo during their estrous cycle with special reference to estrus. Nearly 12 bands have been observed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of whole saliva. Collectively, 179 proteins are identified with respect to different phases of the estrous cycle using mass spectrometry. On the whole, 37 proteins are exclusively expressed in the estrus phase, which include β-enolase, Toll-like receptor (TLR) 4, clusterin, lactoperoxidase, serotransferrin, TGM3, UBA6, and transducin. Among the proteins, β-enolase and TLR 4 were validated, and their specific expression was found during estrus as compared to other phases using immunoblot. The functional annotation reveals many as binding proteins in the estrus saliva when compared to the other phases. The present findings conclude that the proteomic approach adopted to identify the proteins from buffalo saliva around the estrous cycle may provide a new tool for screening the estrus phase. The results further conclude that the specific expression of β-enolase and TLR 4 can be taken as the indicator of estrus in buffalo.—Muthukumar, S., Rajkumar, R., Rajesh, D., Saibaba, G., Liao, C.-C., Archunan, G., Padmanabhan, P., Gulyas, B. Exploration of salivary proteins in buffalo: an approach to find marker proteins for estrus

Structural elucidation of estrus urinary lipocalin protein (EULP) and evaluating binding affinity with pheromones using molecular docking and fluorescence study

Transportation of pheromones bound with carrier proteins belonging to lipocalin superfamily is known to prolong chemo-signal communication between individuals belonging to the same species. Members of lipocalin family (MLF) proteins have three structurally conserved motifs for delivery of hydrophobic molecules to the specific recognizer. However, computational analyses are critically required to validate and emphasize the sequence and structural annotation of MLF. This study focused to elucidate the evolution, structural documentation, stability and binding efficiency of estrus urinary lipocalin protein (EULP) with endogenous pheromones adopting in-silico and fluorescence study. The results revealed that: (i) EULP perhaps originated from fatty acid binding protein (FABP) revealed in evolutionary analysis; (ii) Dynamic simulation study shows that EULP is highly stable at below 0.45 Å of root mean square deviation (RMSD); (iii) Docking evaluation shows that EULP has higher binding energy with farnesol and 2-iso-butyl-3-methoxypyrazine (IBMP) than 2-naphthol; and (iv) Competitive binding and quenching assay revealed that purified EULP has good binding interaction with farnesol. Both, In-silico and experimental studies showed that EULP is an efficient binding partner to pheromones. The present study provides impetus to create a point mutation for increasing longevity of EULP to develop pheromone trap for rodent pest management.Published versio

Melting and solidification of pure metals by a phase-field model

In this paper, we study melting and solidification for metallurgical processes related with phase transitions of pure metals, which during the solid phase show an evident ductility. So, the transition is between a viscous fluid and a viscoplastic state. In this work these particular phenomena can be well described by a phase field fractional model, whose evolution has to satisfy a Ginzburg- Landau equation. Then, we prove the compatibility with the Thermodynamic Laws. Hence, for metallurgical phase transitions, we have considered a similar model by a new fractional derivative and compared the behavior of the first with this second model. Finally, a generalization to finite deformation for the same models is presented in the last section

Buffalo nasal odorant-binding protein (bunOBP) and its structural evaluation with putative pheromones

Pheromones are odoriferous volatile chemical cues produced by animals for communication among conspecifics so as to regulate their social behaviors. In general, the odor compounds are recognized by receptors in the nasal cavity. Odorant-binding protein (OBP), a lipocalin family protein, mediates the air-borne odor cues to nasal receptors through nasal mucus. The presence of OBP in several mammalian species is well documented but to-date there is no report of a nasal OBP in buffalo. Hence, the present study was undertaken to investigate if OBP is present in buffalo nasal mucus. Uni- and two-dimensional gel electrophoresis of the nasal mucus suggested the presence of OBP, which was confirmed using mass spectrometry. In silico homology model of the OBP was generated and its structural similarity with other mammalian OBPs was assessed. Finally, molecular-docking and -dynamics simulations analysis revealed the efficiency of buffalo nasal OBP (bunOBP) to bind with buffalo pheromones as well as other reported chemical cues. Taken together, the occurrence of nasal OBP in buffalo and its putative role in odor binding are reported for the first time. The potential association of this protein with estrus-specific volatiles could be taken to advantage for non-invasive detection of estrus in buffaloes.MOE (Min. of Education, S’pore)Published versio

Intensity (band area) of 14.5 kDa protein around estrous cycle and Ovariectomized animal.

<p>The intensity of the protein band was compared with estrous phases and ovariectomized animal. The protein intensity significantly high during estrus (E) and metestrus (ME) compared to proestrus (PE), diestrus (DE) and ovariectomized (OVX) using Fisher’s least significant difference post-hoc comparisons (*p<0.05). Values are mean ± SE from six gels (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071357#pone.0071357.s002" target="_blank">Figure S2</a>).</p