Krishnaraj Thirugnanasambantham

Swertiamarin attenuates inflammation mediators via modulating NF-κB/I κB and JAK2/STAT3 transcription factors in adjuvant induced arthritis.

Rheumatoid arthritis (RA) is an autoimmune and chronic inflammatory disease that leads to pannus formation followed by severe joint destruction, characterized by synovial hyperplasia, inflammation and angiogenesis. Swertiamarin is a secoiridoid glycoside that is used as an anti-inflammatory compound, mainly found in Enicostema axillare (Lam) A. Raynal, a medicinal plant used in Indian system of traditional medicine. In the present study, the effect of swertiamarin was evlauated in experimental adjuvant arthritis animal model by the estimation of biochemical (paw thickness, lysosomal enzymes, and urinary degradative products) parameters, proinflammatory cytokines and enzymes along with histopathological and radiographic observations. The proteins of phosphorylated NF-κB/IκB and JAK2/STAT3 transcription factors were also quantified from experimental animals as well as LPS induced RAW 264.7 macrophage cells. In in silico analysis, swertiamarin was docked with proinflammatory enzymes to confirm its potential. The administration of swertiamarin (2, 5, 10mg/kg bw) significantly (P⩽0.05) inhibited the levels of paw thickness, lysosomal enzymes and increased the body weight of experimental animals in a dose dependent manner. In molecular analysis, the treatment decreased the release of proinflammatory cytokines (IL1, TNF, IL-6) and proangiogenic enzymes (MMPs, iNOS, PGE2, PPARγ and COX-2); and also significantly (P⩽0.05) increased the levels of antiinflammatory proteins (IL-10, IL-4) when compared to the disease groups. The swertiamarin treatment significantly (P⩽0.05) inhibited the release of NF-κB p65, p-IκBα, p-JAK2 and p-STAT3 signaling proteins levels on both experimental animals and LPS induced cells. Histopathological and radiological analysis evidenced the curative effect of swertiamarin on bone destruction. The docking studies of swertiamarin on proinflammatory enzymes supported the results from the in vivo experiments. Thus the swertiamarin inhibited the development of arthritis by modulating NF-κB/IκB and JAK2/STAT3 signaling. These findings suggested that swertiamarin acted as an anti-rheumatic agent.

Relevance of miR-21 in HIV and non-HIV-related lymphomas

The critical role of microRNAs (miRNAs) in cell differentiation, homeostasis and cancer development has been extensively discussed in recent publications. The microRNAs with RISC enzyme complex allow it to find its complementary sequence, which is usually located in the 3′-untranslated region (UTR) of the target messenger RNA (mRNA). This is followed by inhibition of protein translation or promotion, resulting in degradation of the target gene. miR-21 has been mapped at chromosome 17q23.2, where it overlaps with the protein coding gene vacuole membrane protein 1 (VMP1), a human homologue of rat vacuole membrane protein. Recent evidence indicates that miR-21 plays a vital role in tumour cell proliferation, apoptosis and invasion. The inhibition of miR-21 may induce cell cycle arrest and increased chemosensitivity to anticancer agents, providing evidence that miR-21 functions as an oncogene in human cancer. Increased expression levels of miR-21 were observed in tumours arising from diverse tissue types. This also includes tumours of haematological origin, such as chronic lymphatic leukaemia, diffuse large B cell lymphomas (DLBCLs), acute myeloid leukaemia and Hodgkin lymphomas. Recently, it has been shown that high levels of B cell activation were induced by miR-21 in circulating B cells and are seen in HIV-infected individual. Notably, miR-21 is overexpressed in activated B cells, suggesting its assistance in maintaining B cell hyperactivation, which plays a pivotal role in HIV-infected cells. Therefore, miR-21 can be considered as a powerful biomarker in HIV-related lymphomas. The number of studies related to the role of miR-21 in HIV-related lymphomas is sparse; therefore, this mini review highlights the recent publications related to clinical impact and significance of miR-21, specifically in HIV- and non-HIV-related lymphomas.

Role of Ethylene Response Transcription Factor (ERF) and Its Regulation in Response to Stress Encountered by Plants

Plants are nonmotile and are easily affected by both biotic and abiotic stresses. Plants have evolved themselves at both cellular and molecular level to fight against stress. Transcription factors are important among the stress-responsive genes, and their protein products are known to regulate the expression of other stress-responsive genes via binding to the regulatory elements. Among the plant transcription factors, ethylene response factor (ERF) is one of the largest subfamilies of Apetala2 (AP2)/ERF transcription factor family and is characterized with single AP2 domain. ERFs are a double-edged sword; though most of the ERFs are activators of stress-responsive genes, certain ERF could act as repressor, and this phenomenon of ERF has been well discussed in this review. Further, the expression of ERFs may be ethylene dependent or independent and is regulated by feedback mechanism. Apart from above regulation mechanism, expressions of ERFs are post-transcriptionally regulated by microRNAs (miRNAs), and miRNA expressions are in turn regulated by ERFs. The present review highlights the importance of ERFs in plant stress management and complexity in regulation of ERF expression in response to various stresses.

Novel Aryl Hydrocarbon Receptor Agonist Suppresses Migration and Invasion of Breast Cancer Cells

Background Despite the remarkable progress to fight against breast cancer, metastasis remains the dominant cause of treatment failure and recurrence. Therefore, control of invasiveness potential of breast cancer cells is crucial. Accumulating evidences suggest Aryl hydrocarbon receptor (Ahr), a helix-loop-helix transcription factor, as a promising target to control migration and invasion in breast cancer cells. Thus, an Ahr-based exploration was performed to identify a new Ahr agonist with inhibitory potentials on cancer cell motility. Methods For prediction of potential interactions between Ahr and candidate molecules, bioinformatics analysis was carried out. The interaction of the selected ligand with Ahr and its effects on migration and invasion were examined in vitro using the MDA-MB-231 and T47D cell lines. The silencing RNAs were transfected into cells by electroporation. Expressions of microRNAs (miRNAs) and coding genes were quantified by real-time PCR, and the protein levels were detected by western blot. Results The in silico and in vitro results identified Flavipin as a novel Ahr agonist. It induces formation of Ahr/Ahr nuclear translocator (Arnt) heterodimer to promote the expression of cytochrome P450 family 1 subfamily A member 1 (Cyp1a1). Migration and invasion of MDA-MB-231 and T47D cells were inhibited with Flavipin treatment in an Ahr-dependent fashion. Interestingly, Flavipin suppressed the pro-metastatic factor SRY-related HMG-box4 (Sox4) by inducing miR-212/132 cluster. Moreover, Flavipin inhibited growth and adhesion of both cell lines by suppressing gene expressions of B-cell lymphoma 2 (Bcl2) and integrinα4 (ITGA4). Conclusion Taken together, the results introduce Flavipin as a novel Ahr agonist, and provide first evidences on its inhibitory effects on cancer cell motility, suggesting Flavipin as a candidate to control cell invasiveness in breast cancer patients.

Pinocembrin, a novel histidine decarboxylase inhibitor with anti-allergic potential in in vitro

ABSTRACT Pinocembrin (5, 7‐ dihydroxy flavanone) is the most abundant chiral flavonoid found in propolis, exhibiting antioxidant, antimicrobial and anti‐inflammatory properties. However, the effect of Pinocembrin on allergic response is unexplored. Thus, current study aimed at investigating the effects of Pinocembrin on IgE‐mediated allergic response in vitro. A special emphasis was directed toward histidine decarboxylase (HDC) and other pro‐allergic and pro‐inflammatory mediators. Preliminary studies, using a microbiological model of Klebsiella pneumoniae, provided first evidences that suggest Pinocembrin as a potential thermal stable inhibitor for HDC. Applying docking analysis revealed possible interaction between Pinocembrin and mammalian HDC. In vitro studies validated the predicted interaction and showed that Pinocembrin inhibits HDC activity and histamine in IgE‐sensitized RBL‐2H3 in response to dinitrophenol (DNP)‐bovine serum albumin (BSA) stimulation. In addition, Pinocembrin mitigated the damage in the mitochondrial membrane, formation of cytoplasmic granules and degranulation as indicated by lower &bgr;‐hexoseaminidase level. Interestingly, it reduced range of pro‐inflammatory mediators in the IgE‐mediated allergic response including tumor necrosis factor (TNF)‐&agr;, interleukin (IL)‐6, nitric oxide (NO), inducible NO synthase (iNOS), phosphorylation of inhibitory kappa B (I&kgr;B)‐&agr;, prostaglandin (PGE)‐2 and cyclooxygenase (COX)‐2. In conclusion, current study suggests Pinocembrin as a potential HDC inhibitor, and provides the first evidences it is in vitro anti‐allergic properties, suggesting Pinocembrin as a new candidate for natural anti‐allergic drugs.

Swertiamarin, a natural steroid, prevent bone erosion by modulating RANKL/RANK/OPG signaling

Abstract Bone erosion is a central feature of rheumatoid arthritis (RA) that is characterized by the infiltration of the synovial lining by osteoclasts and lymphocytes. In the present study, swertiamarin a major secoiridoid glycoside was evaluated for anti‐osteoclastogenic property to prevent bone erosion in Freunds complete adjuvant (FCA) induced in‐vivo model, in‐vitro osteoblast and osteoclasts as well as in co‐culture system and in‐silico molecular docking analysis. The swertiamarin treatment decreased the expression of TRAP, RANKL, and RANK levels and increased the levels of OPG levels significantly in both in vitro and in vivo models. In in vitro, the compound treatment significantly increased the cell proliferation and ALP levels in osteoblast cells; the high proliferation (153.8600 ± 5.23%) and ALP release (165.6033 ± 4.13%) were observed at 50 &mgr;g/ml concentration of swertiamarin treatment. At the same time the treatment decreased the TRAP positive cells in osteoclast cells; the high reductions of TRAP positive cells (39.32 ± 3.19%) were observed at 50 &mgr;g/ml of swertiamarin treatment. The treatment modulated the levels of pro‐inflammatory cytokines, MMPs and NF‐&kgr;B levels in osteoblast and osteoclast co‐culture system. In in silico analysis swertiamarin had affinity towards the proteins RANK, RANKL and OPG residues with low binding energy −4.5, −3.92 and −5.77 kcal/mol respectively. Thus, the results of this study revealed the anti‐osteoclastogenic activity of swertiamarin on the prevention of bone destruction. HighlightsSwertiamarin was confirmed as anti‐osteoclastogenic effect by in‐silico analysis on bone mediated proteins.The swertiamarin reciprocally regulate the expression of TRAP, RANKL, RANK levels and OPG levels.The swertiamarin modulated the levels of pro‐inflammatory cytokines, MMPs and NF‐&kgr;B levels.Swertiamarin treatment revealed the prevention of bone destruction in animal models.

In Silico Identification of Human miR 3654 and its Targets Revealed its Involvement in Prostate Cancer Progression

BACKGROUND MicroRNAs (miRNAs) are non-coding RNAs known to control a broad range of biological functions such as cellular proliferation, differentiation and programmed cell death. Recent reports showed that miRNAs can act as oncogenes or tumor suppressors, thereby, playing an important role in cancer initiation and progression. Moreover, we know that Expressed sequence tags (ESTs) are random single pass sequence reads, which displays the condition/tissue specific transcripts (coding and non-coding) of an organism. METHODS In the present study, we have applied the bioinformatics approach to identify miRNA from prostate cancer using EST resource and its expressions were analyzed by quantitative reverse transcription PCR (qRT-PCR). RESULTS Analysis of transcriptomics resource from the LNCaP cells revealed the presence of an EST encoding hsa-miR-3654. Presence of the premature candidate of miR-3654, demonstrates its expression in LNCaP cells. We further indentified that the expression level (Fold Induction) of miR-3654 in LNCaP was higher than the normal and androgen insensitive prostate cancer cell lines (PNT1A, PC-3). CONCLUSION we have identified the miR-3654 involved in prostate cancer progression using computational approach and hypothesized that the down regulation of miR-3654 could be responsible for a solid tumor to get cancer stem-like cell phenotype. Further studies are required to investigate the molecular mechanisms behind the STAT3 mediated miR-3654 repression and the associated metastasis.

Sequencing approaches in cancer treatment.

Use of sequencing approaches is an important aspect in the field of cancer genomics, where next‐generation sequencing has already been utilized for targeting oncogenes or tumour‐suppressor genes, that can be sequenced in a short time period. Alterations such as point mutations, insertions/deletions, copy number alterations, chromosomal rearrangements and epigenetic changes are encountered in cancer cell genomes, and application of various NGS technologies in cancer research will encounter such modifications. Rapid advancement in technology has led to exponential growth in the field of genomic analysis. The

Computational Approach for Identification of Anopheles gambiae miRNA Involved in Modulation of Host Immune Response

1000 Genome Project (in which the goal is to sequence an entire human genome for

Analysis of Dormant Bud (Banjhi) Specific Transcriptome of Tea (Camellia sinensis (L.) O. Kuntze) from cDNA Library Revealed Dormancy-Related Genes

1000), and deep sequencing techniques (which have greater accuracy and provide a more complete analysis of the genome), are examples of rapid advancements in the field of cancer genomics. In this mini review, we explore sequencing techniques, correlating their importance in cancer therapy and treatment.