RF-amide related peptide-3 (RFRP-3): a novel neuroendocrine regulator of energy homeostasis, metabolism, and reproduction.

A hypothalamic neuropeptide, RF-amide related peptide-3 (RFRP-3), the mammalian ortholog of the avian gonadotropin-inhibitory hormone (GnIH) has inhibitory signals for reproductive axis via G-protein coupled receptor 147 in mammals. Moreover, RFRP-3 has orexigenic action but the mechanism involved in energy homeostasis and glucose metabolism is not yet known. Though, the RFRP-3 modulates orexigenic action in co-operation with other neuropeptides, which regulates metabolic cues in the hypothalamus. Administration of GnIH/RFRP-3 suppresses plasma luteinizing hormone, at the same time stimulates feeding behavior in birds and mammals. Likewise, in the metabolically deficient conditions, its expression is up-regulated suggests that RFRP-3 contributes to the integration of energy balance and reproduction. However, in many other metabolic conditions like induced diabetes and high-fat diet obesity, etc. its role is still not clear while, RFRP-3 induces the glucose homeostasis by adipocytes is reported. The physiological role of RFRP-3 in metabolic homeostasis and the metabolic effects of RFRP-3 signaling in pharmacological studies need a detailed discussion. Further studies are required to find out whether RFRP-3 is associated with restricted neuroendocrine function observed in type II diabetes mellitus, aging, or sub-fertility. In this context, the current review is focused on the role of RFRP-3 in the above-mentioned mechanisms. Studies from search engines including PubMed, Google Scholar, and science.gov are included after following set inclusion/exclusion criteria. As a developing field few mechanisms are still inconclusive, however, based on the available information RFRP-3 seems to be a putative tool in future treatment strategies towards metabolic disease.

Heavy Metals Assessment in Water, Sediments, Algae and Two Fish Species from River Swat, Pakistan.

The heavy metals were studied in water, sediments, algae, and various tissues of Glyptosternon reticulatum and Cyprinus carpio from River Swat, Pakistan, using flame atomic absorption spectrophotometer. The Zn, Cu, Pb and Ni were higher in water at sewage site compared to upstream and downstream sites. In sediments, the Ni and Cd were not detected whereas Cu, Pb and Zn were higher at downstream followed by sewage and upstream sites. The Ni and Zn in algae were higher at upstream and sewage sites compared to downstream site whereas Pb and Cd were higher at upstream site compared to sewage and downstream sites and Cu was found same at all the three sites. The heavy metals (Zn > Cu > Pb and Ni) in tissues (liver > gills > skin > muscles) of G. reticulatum was higher than in C. carpio. This study recommends the proper monitoring of River Swat in order to save its water and inhabitant aquatic life.

Rosiglitazone Enhances Browning Adipocytes in Association with MAPK and PI3-K Pathways During the Differentiation of Telomerase-Transformed Mesenchymal Stromal Cells into Adipocytes.

Obesity is a major risk for diabetes. Brown adipose tissue (BAT) mediates production of heat while white adipose tissue (WAT) function in the storage of fat. Roles of BAT in the treatment of obesity and related disorders warrants more investigation. Peroxisome proliferator activator receptor gamma (PPAR-γ) is the master regulator of both BAT and WAT adipogenesis and has roles in glucose and fatty acid metabolism. Adipose tissue is the major expression site for PPAR-γ. In this study, the effects of rosiglitazone on the brown adipogenesis and the association of MAPK and PI3K pathways was investigated during the in vitro adipogenic differentiation of telomerase transformed mesenchymal stromal cells (iMSCs). Our data indicate that 2 µM rosiglitazone enhanced adipogenesis by over-expression of PPAR-γ and C/EBP-α. More specifically, brown adipogenesis was enhanced by the upregulation of EBF2 and UCP-1 and evidenced by multilocular fatty droplets morphology of the differentiated adipocytes. We also found that rosiglitazone significantly activated MAPK and PI3K pathways at the maturation stage of differentiation. Overall, the results indicate that rosiglitazone induced overexpression of PPAR-γ that in turn enhanced adipogenesis, particularly browning adipogenesis. This study reports the browning effects of rosiglitazone during the differentiation of iMSCs into adipocytes in association with the activation of MAPK and PI3K signaling pathways.

Dietary administration of the commercially available probiotics enhanced the survival, growth, and innate immune responses in Mori (Cirrhinus mrigala) in a natural earthen polyculture system.

The use of probiotics is considered effective for survival, growth and enhanced immune response in aquaculture. In the current study, effects of commercially available probiotic (Magic Plus) was investigated on survival, growth and immune response of Mori (Cirrhinus mrigala) in a polyculture system. The experiment was conducted for 90 days on 1200 fingerlings in two groups i.e. control and probiotic supplemented groups each having 600 fingerlings. Control group was fed with 35% protein basal diet without any supplements and the other group was supplemented with commercially available probiotic at the rate of (1012 CFU kg-1 diet). After 90 days, probiotic supplemented group was characterized with significant increase (p < 0.05) in growth parameters like, total weight, total length, %weight gain, specific growth rate and survival growth rate. Immunological indices like, lysozyme activity, white blood cells, total plasma protein level and immunoglobulin (IgM) of supplemented group were also significantly (p < 0.05) enhanced. Moreover, digestive enzymes i.e. cellulase, protease and amylase were also found to be significantly (p < 0.05) hyper-active in probiotic supplemented groups. Haematological parameters like, RBCs, Hb, Hct, MCH and MCHC were also significantly (p < 0.05) increased. Thus, the current study strongly suggests that a commercially available probiotic Magic plus may serve as a healthy and immunostimulating feed additive in C. mrigala culture.

Cyclophilin A: A Key Factor in Virus Replication and Potential Target for Anti-viral Therapy.

Cyclophilin A (CypA) is a key member of immunophilins that has peptidyl-prolyl cis/trans isomerase (PPIase) activity. Besides acting as a cellular receptor for immunosuppressive drug cyclosporine A (CsA), CypA is involved in various cellular activities. CypA has an important role in viral infection which either facilitates or inhibits their replication. Inhibition of CypA via inhibitors is useful for overcoming several viral infections, indicating that CypA is an attractive target for anti-viral therapy. Collectively, these facts demonstrate the critical roles of CypA in mediating or inhibiting viral infections, suggesting that CypA can be an attractive cellular target for the development of anti-viral therapy.

Factors affecting sustainable iodine deficiency elimination in Pakistan: A global perspective.

Iodine deficiency remains a considerable challenge worldwide, even after decades of efforts to address the problem. The aim of this review is to present the current situation in historically iodine-deficient Pakistan regarding iodine nutritional status and place it in a global perspective. We collected relevant articles from online bibliographic databases and websites of concerned organizations that addressed prevalence of goiter/iodine deficiency and barriers to sustainable control. We divided the studies into pre- and post-1994, a landmark year when Pakistan formally adopted the universal salt iodization (USI) programme. Overall, 56 studies reported goiter/iodine deficiency prevalence in Pakistan. Before 1994, six studies (30%) reported a goiter prevalence ≥70%, while nine studies (45%) reported a goiter prevalence between 30% and 70%. Only five studies (25%) found a goiter prevalence less than 30%, of which only two studies reported prevalence <10%. From 1994 onwards, 15 studies (41.7%) reported a goiter/iodine deficiency (ID) prevalence ≥50%, of which seven studies reported prevalence ≥70%, while three studies (8.3%) found a goiter prevalence of 30%-49%, nine studies (25%) found a goiter prevalence of 10%-29%, and five studies (13.9%) reported prevalence of <10%. Four studies (11.1%) reported lower goiter prevalence but higher prevalence of iodine deficiency. The efforts in the past two decades resulted in up to a 50% decline in iodine deficiency disorders (IDD). Variable remaining factors and the recent results, however, indicate that this decline may be non-uniform and even over-estimated. Coordinated and regionally adopted efforts for eradication of IDD from all stakeholders should be pursued. Policy makers should take steps to protect future generations and alert concerned organizations about the importance of careful assessments and estimates of iodine nutritional status.

Decreased fish diversity found near marble industry effluents in River Barandu, Pakistan.

In a recently published study we observed that effluents from marble industry affected physicochemical characteristics of River Barandu in District Buner, Pakistan. These changes in water quality due to marble effluents may affect fish community. The present study was therefore conducted to evaluate the impacts of marble industry effluents on fish communities in River Barandu using abundance, richness, diversity and evenness of fish species as end point criteria. The fish samples were collected by local fishermen on monthly basis from three selected sites (upstream, effluents/industrial, and downstream sites). During the study period, a total of 18 fish species were found belonging to 4 orders, 5 families and 11 genera. The Cyprinidae was observed to be the dominant family at all the three selected sites. Lower abundance and species diversity was observed at the industrial (22%) and downstream sites (33%) as compared to the upstream site (45%). Effluents of marble industry were associated with lower abundance of species in River Barandu. It is recommended that industries should be shifted away from the vicinity of river and their effluents must be treated before discharging to prevent further loss of fish abundance and diversity in the River.

Chemotactic Activity of Cyclophilin A in the Skin Mucus of Yellow Catfish (Pelteobagrus fulvidraco) and Its Active Site for Chemotaxis.

Fish skin mucus is a dynamic barrier for invading pathogens with a variety of anti-microbial enzymes, including cyclophilin A (CypA), a multi-functional protein with peptidyl-prolyl cis/trans isomerase (PPIase) activity. Beside various other immunological functions, CypA induces leucocytes migration in vitro in teleost. In the current study, we have discovered several novel immune-relevant proteins in yellow catfish skin mucus by mass spectrometry (MS). The CypA present among them was further detected by Western blot. Moreover, the CypA present in the skin mucus displayed strong chemotactic activity for yellow catfish leucocytes. Interestingly, asparagine (like arginine in mammals) at position 69 was the critical site in yellow catfish CypA involved in leucocyte attraction. These novel efforts do not only highlight the enzymatic texture of skin mucus, but signify CypA to be targeted for anti-inflammatory therapeutics.

Identification and Characterization of MicroRNAs in Snakehead Fish Cell Line upon Snakehead Fish Vesiculovirus Infection.

MicroRNAs (miRNAs) play important roles in mediating multiple biological processes in eukaryotes and are being increasingly studied to evaluate their roles associated with cellular changes following viral infection. Snakehead fish Vesiculovirus (SHVV) has caused mass mortality in snakehead fish during the past few years. To identify specific miRNAs involved in SHVV infection, we performed microRNA deep sequencing on a snakehead fish cell line (SSN-1) with or without SHVV infection. A total of 205 known miRNAs were identified when they were aligned with the known zebrafish miRNAs, and nine novel miRNAs were identified using MiRDeep2 software. Eighteen and 143 of the 205 known miRNAs were differentially expressed at three and 24 h post-infection (poi), respectively. From the differentially-expressed miRNAs, five were randomly selected to validate their expression profiles using quantitative reverse transcription polymerase chain reaction (qRT-PCR), and their expression profiles were consistent with the microRNA sequencing results. In addition, the target gene prediction of the SHVV genome was performed for the differentially-expressed host miRNAs, and a total of 10 and 58 differentially-expressed miRNAs were predicted to bind to the SHVV genome at three and 24 h poi, respectively. The effects of three selected miRNAs (miR-130-5p, miR-214 and miR-216b) on SHVV multiplication were evaluated using their mimics and inhibitors via qRT-PCR and Western blotting. The results showed that all three miRNAs were able to inhibit the multiplication of SHVV; whereas the mechanisms underlying the SHVV multiplication inhibited by the specific miRNAs need to be further characterized in the future.

Impact of marble industry effluents on water and sediment quality of Barandu River in Buner District, Pakistan.

Industries play an important role in improving the living standard but at the same time cause several environmental problems. Therefore, it is necessary to evaluate the impact of industries on the quality of environment. In the present study, the impact of marble industry effluents on water and sediment quality of Barandu River in Buner District, Pakistan was evaluated. Water and sediment samples were collected at three different sampling sites (upstream, industrial, and downstream sites) from Barandu River and their physicochemical properties were inter-compared. In addition, different marble stones and mix water (wastewater) from marble industry were analyzed. The measured physicochemical parameters of river water including pH, electrical conductivity (EC), alkalinity, total hardness, Ca and Mg hardness, total dissolved solid (TDS), total suspended solids (TSS), sulfates (SO4 (2-)), sodium (Na(+)), potassium (K(+)), nitrites (NO2 (-)), nitrate (NO3 (-)), chloride (Cl(-)), calcium (Ca(2+)), and magnesium (Mg(2+)) were found to be significantly altered by effluent discharges of marble industries. Similarly, heavy metal concentrations in both water and sediments of the river were significantly increased by marble industry wastewater. It is concluded that large quantities of different pollutants are added to Barandu River due to direct disposal of marble industry effluents which degrades its quality. Therefore, it is recommended that direct disposal of marble industry wastewater should be banned and all effluents must be properly treated before discharging in the river water.

Determination of heavy metal contents in water, sediments, and fish tissues of Shizothorax plagiostomus in river Panjkora at Lower Dir, Khyber Pakhtunkhwa, Pakistan.

The present study was conducted to investigate the contamination of water, sediments, and fish tissues with heavy metals in river Panjkora at Lower Dir, Khyber Pakhtunkhwa, Pakistan. Water, sediments, and fish (Shizothorax plagiostomus) samples were collected from September 2012 to January 2013 at three different sites (upstream site at Sharigut, sewage site at Timergara, and downstream site at Sadoo) of river Panjkora. The concentrations of heavy metals in water were in the order Zn > Cu ≈ Pb > Ni ≈ Cd with mean values of 0.30, 0.01, 0.01, 0.0 and 0.0 mg/l, respectively, which were below the maximum permissible limits of WHO for drinking water. In sediments, heavy metals were found in the order Cu > Zn > Ni > Pb > Cd with mean concentrations of 50.6, 38.7, 9.3, 8, and 0.4 mg/kg, respectively. Ni and Cd were not found in any fish tissues, but Zn, Cu, and Pb were detected with the mean concentration ranges of 0.04-1.19, 0.03-0.12, and 0.01-0.09 µg/g, respectively. The present study demonstrates that disposal of waste effluents causes a slight increase in the concentration of heavy metals in river Panjkora as revealed by variation in metal concentrations from upstream to downstream site. Sewage disposal was also found to change physicochemical characteristics of Panjkora water. At present, water and fish of river Panjkora are safe for human consumption, but the continuous sewage disposal may create problems in the future.

Central obesity and fat-free mass are associated with a larger spleen volume in the general population.

Background and aim: As the spleen plays a significant role in immunity, the aim was to investigate the associations of different body composition markers derived from various sources with spleen volume in a general population sample. Materials and methods: Cross-sectional data of 1095 individuals (570 women; 52%) aged between 30 and 90 years were collected in the Study of Health in Pomerania (SHIP-START-2). We measured spleen volume by magnetic resonance imaging (MRI).Body composition markers were derived from classic anthropometry, bioelectrical impedance analysis, including absolute fat mass (FM) and fat-free mass (FFM), as well as from MRI, including visceral adipose tissue (VAT), subcutaneous adipose tissue (SAT), and liver fat content. Sex-stratified-adjusted linear regression models were used to analyze the associations of body composition markers with spleen volumes. Results: We observed positive associations of body mass index, body weight, waist circumference, hip circumference, waist-to-height ratio, absolute FM, absolute FFM, and VAT and SAT with spleen volume in men and women. An 8.12 kg higher absolute FFM was associated with a 38.4 mL (95% confidence interval [CI]: 26.7-50.1) higher spleen volume in men and a 5.21 kg higher absolute FFM with a 42.6 mL (95% CI: 26.2-59.0) higher spleen volume in women. Conclusion: Our findings indicate that obesity-related body composition markers and FFM are associated with a higher spleen volume. Particularly, higher absolute FFM showed a strong association with a larger spleen volume in both men and women. Further studies are warranted to understand the clinical significance of body composition markers on large spleen volume.

Proteomic profile of epidermal mucus from Labeo rohita reveals differentially abundant proteins after Aeromonas hydrophila infection.

We report the proteomic profile of Epidermal Mucus (EM) from Labeo rohita and identified the differentially abundant proteins (DAPs) against Aeromonas hydrophila infection through label-free liquid chromatography-mass spectrometry (LC-MS/MS). Using discovery-based proteomics, a total of 2039 proteins were quantified in nontreated group and 1,328 proteins in the treated group, of which 114 were identified as DAPs in both the groups. Of the 114 DAPs, 68 proteins were upregulated and 46 proteins were downregulated in the treated group compared to nontreated group. Functional annotations of these DAPs shows their association with metabolism, cellular process, molecular process, cytoskeletal, stress, and particularly immune system. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis and Fisher's exact test between the two groups shows that most of the proteins were immune-related, which were significantly associated with the proteasome, phagosome, and Salmonella infection pathways. Overall, this study shows a basic and primary way for further functional research of the involvement of vitellogenin 2, alpha-2-macroglobulin-like protein, toll-like receptors (TLR-13), calpain, keratin-like proteins, and heat shock proteins against bacterial infection. Nonetheless, this first-ever comprehensive report of a proteomic sketch of EM from L. rohita after A. hydrophila infection provides systematic protein information to broadly understand the biological role of fish EM against bacterial infection.

First Record of Ichthyofauna from Gomal Zam Dam, District South Waziristan, Khyber Pakhtunkhwa, Pakistan.

The present study reports the fish fauna of Gomal Zam Dam at three different sites from October 2018 to July 2019. The total sampled fish belong to 18 species, 15 genera, 5 families, and 4 orders. Fourteen species belong to family Cyprinidae, which was the most abundant, while one species belong to each family Mastacembelidae, Channidae, Siluridae, and Sisoridae. The prominent species were Labeo dyochielus (18.6%), Barilius vagra (16.5%), and Barilius pakistanicus (13.8%), while the rare species were Tor zhobensis, Wallago attu, Hyphophthalmicthys molitrix, Ctenophyrngodon idella, and Bagarius bagarius each one forming (0.1%). Species were rich in spring with ideal temperature, followed by summer, whereas species abundance was high in summer with high temperature and minimum abundance was recorded in autumn. In conclusion, the Gomal Zam Dam is a favorable reservoir for the fish particularly for family Cyprinidae. This present study will provide useful information about the diversity of fish fauna of Gomal Zam Dam that could be used in systematic fisheries management and conservation of the country.

Increased strength in the Col-Tgel induces apoptosis in the human dental pulp stem cells: 3D culturing of human dental pulp stem cells at different strengths of collagen.

Human dental pulp stem cells (HDPSCs) have great potential to be used in regenerative medicine. To use these stem cells effectively for this purpose, they should be grown in a 3D cell culture that mimics their natural niches instead of a 2D conventional cell culture. The aim of this study was to grow the HDPSCs in the 3D cell culture created by Transglutaminase-crosslinked collagen hydrogels (Col-Tgel) in two different strengths to find a suitable 3D cell culture environment for these stem cells. Two stiffness of the 3D Col-Tgel were used to grow the HDPSCs: soft and medium matrix with strength of 0.9-1.5 kPa and 14-20 kPa, respectively. HDPSCs express markers similar to MSCs, therefore seven such markers were analyzed in the HDPSCs during their growth in the 2D and in the 3D soft and medium Col-Tgel. The CD105 and CD90 markers were significantly (p < 0.05) downregulated in HDPSCs cultured in both 3D cell culture conditions compared with HDPSCs in 2D cell culture. Furthermore, CD34 marker, a negative marker, expressed by a few cells in HDPSCs culture was upregulated (p < 0.05) in HDPSCs cultured in medium 3D Col-Tgel, indicating cells that expressing the marker grow better in medium 3D Col-Tgel. The apoptosis results revealed that HDPSCs in medium 3D Col-Tgel had the least number of live cells and a significantly (p < 0.05) higher early apoptosis rate compared to HDPSCs in 2D and 3D Col-Tgel medium. MTT analysis also showed a significant difference among the three cell culture conditions. We conclude that HDPSCs cultured on 3D soft Col-Tgel showed better proliferation than cells cultured in 3D medium gel. These results demonstrate that the ideal environment to grow HDPSCs in 3D is the soft Col-Tgel not medium Col-Tgel.

Transcriptomic profiling of the telomerase transformed Mesenchymal stromal cells derived adipocytes in response to rosiglitazone.

BACKGROUND: Differentiation of Immortalized Human Bone Marrow Mesenchymal Stromal Cells - hTERT (iMSC3) into adipocytes is in vitro model of obesity. In our earlier study, rosiglitazone enhanced adipogenesis particularly the brown adipogenesis of iMSC3. In this study, the transcriptomic profiles of iMSC3 derived adipocytes with and without rosiglitazone were analyzed through mRNA sequencing. RESULTS: A total of 1508 genes were differentially expressed between iMSC3 and the derived adipocytes without rosiglitazone treatment. GO and KEGG enrichment analyses revealed that rosiglitazone regulates PPAR and PI3K-Akt pathways. The constant rosiglitazone treatment enhanced the expression of Fatty Acid Binding Protein 4 (FABP4) which enriched GO terms such as fatty acid binding, lipid droplet, as well as white and brown fat cell differentiation. Moreover, the constant treatment upregulated several lipid droplets (LDs) associated proteins such as PLIN1. Rosiglitazone also activated the receptor complex PTK2B that has essential roles in beige adipocytes thermogenic program. Several uniquely expressed novel regulators of brown adipogenesis were also expressed in adipocytes derived with rosiglitazone: PRDM16, ZBTB16, HOXA4, and KLF15 in addition to other uniquely expressed genes. CONCLUSIONS: Rosiglitazone regulated several differentially regulated genes and non-coding RNAs that warrant further investigation about their roles in adipogenesis particularly brown adipogenesis.

Comparative anticancer activities of Ficus carica and Ficus salicifolia latex in MDA-MB-231 cells.

Ficus latex is rich in polyphenolic compounds and hence considered as an antioxidant and anti-proliferative. Many studies are available on Ficus carica (common fig) whereas Ficus salicifolia is less studied. F. salicifolia grows in a harsh dry environment, therefore its latex was selected in the current study along with the F. carica for their comparative anti-cancer potential and the involved molecular mechanism. Triple-negative breast cancer (TNBC) derived MDA-MB-231 cells were used in the study. MTT and morphological studies indicated that the latex of both plants has anti-proliferative effects. To know their anti-metastatic effects, a wound-healing assay was performed. Both species were able to maintain the wound size compared to the untreated cells indicating their anti-metastatic effects. Using a DNA damage assay kit, we found that both fig species have genotoxic and cytotoxic effects in MDA-MB-231 cells compared to the untreated control. To know the potential molecular mechanism involved, we used a human kinase array kit. We found that ERK2, CREB, and AKT2 were downregulated after treatment the MDA-Mb-231 cells with the latex of F. carica. We assumed that F. salicifolia will also affect the same pathways, however after confirmation through real-time (RT)-PCR, downregulations of the above mentioned pathways were confirmed in cells treated with F. carica latex, however, in cells treated with F. salicifolia the selected genes were upregulated at the transcriptional level. We conclude that latex of both species of ficus have anti-cancer effects in MDA-MB-231 cells, however differ in their level of toxicity and the mechanism of action at the molecular level.

Significant transcriptomic changes are associated with the inhibitory effects of 5-aza-2-deoxycytidine during adipogenic differentiation of MG-63 cells.

Our previous study revealed that the treatment of 5-aza-2-deoxycytidine (5-aza) inhibited while treatment of suberoylanilide hydroxamic acid (SAHA) enhanced the adipogenic differentiation of MG-63 cells. In this study, we examined the transcriptomic profiles of the derived adipocyte-like cells from MG-63 cells in the presence of 5-aza (Treatment 1) and SAHA (Treatment 2). Genome wide expression analysis showed high within sample variability for the adipocytes derived with 5-aza versus vehicle. Additionally, the expression profile of 5-aza derived cells was separated from the other sample groups. Differential analysis on the pairwise comparison of 5-aza versus control and SAHA versus 5-aza identified 1290 and 1086 differentially expressed (DE) genes, respectively. Furthermore, some overlap was observed between the up and down-regulated DE genes of 5-aza versus control and SAHA versus control (jaccard score 0.3) as well as between the differentially regulated genes of 5-aza versus control and 5-aza versus SAHA (jaccard score 0.29). A total of 73 transcription factors (TFs) were differentially expressed across all the pair wise comparisons with some overlap between the under and over expressed TFs of 5-aza versus control and 5-aza versus SAHA (jaccard score 0.29). Unsupervised clustering of TFs showed that the samples within the group are consistent in expression and the samples cluster in accordance with the group. Several GO terms related to enhanced adipogenesis such as neutral lipid biosynthetic process, lipid metabolic processes, cellular amide metabolic processes and cellular carbohydrate metabolic processes were enriched in the down regulated genes of 5-aza derived adipocytes versus control, indicating 5-aza inhibit the adipogenic differentiation of MG-63 cells. GSEA analysis on selected gene sets of MAPK and PI3K signaling pathway in MSigDB identified the pathways were up-regulated in 5-aza versus control. This study revealed that inhibition of MG-63 adipogenesis due to 5-aza treatment is associated with large transcriptomics changes and further research is needed to unravel the roles of these genes in the adipogenesis.

Phytochemical profile and antiproliferative activities of acetone extracts of Asplenium polypodioides Blume. and A. dalhousiae Hook. in MDA-MB-231 breast cancer cells.

The plants extracts are widely used in traditional medicines and hence considered a potential source for drug discovery. In this study, we assessed the phytochemical composition of Asplenium dalhousiae and Asplenium polypodioides in acetone extracts and checked its antiproliferative potential in MDA-MB-231 cells. We found that both plants are rich in phenolic and flavonoid compounds and are efficient in antioxidant activities. The total phenolic compounds in A. dalhousiae were 44.15 ± 1.38 µg/mg whereas in A. polypodioides were 27.73 ± 1.35 µg/mg. Total flavonoids in A. dalhousiae were 105.39 ± 2.92 µg/mg whereas in A. polypodioides were 101.56 ± 1.75 µg/mg. The ferric reducing power assay indicates 66.38 ± 2.6% reduction by A. dalhousiae whereas 78.43 ± 0.47% reduction by A. polypodioides. Similarly, the total antioxidant capacity of A. dalhousiae was found to be 59.95 ± 1.13 whereas for A. polypodioides the recorded value was 33.03 ± 1.67%. Using GCMS analysis, we identified 25 compounds in A. dalhousiae whereas 26 in A. polypodioides. Four of these compounds are common in both plants. The morphological study and MTT assay revealed that both plants have antiproliferative potential as both plants exerted significant effects on the shape of the MDA-MB-231 cells and inhibited cellular proliferation in time and dose dependent manner. We conclude that both Asplenium plants have potential anticancer compounds.

Arylated gold nanoparticles have no effect on the adipogenic differentiation of MG-63 cells nor regulate any key signaling pathway during the differentiation.

OBJECTIVE: MG-63 cells that have osteoblastic and adipogenic differentiation potential were evaluated for internalization, and adipogenic differentiation in the presence and absence of the covalently functionalized aryl gold nanoparticles (AuNPs-C6H4-4-COOH). RESULTS: Inductively coupled plasma, flow cytometry and confocal microscopy analyses confirmed that gold nanoparticles were easily internalized by MG-63 cells. The MG-63 cells were differentiated into adipocytes without gold-aryl nanoparticles and with the gold-aryl nanoparticles at 5 µM concentration in both induction and maintenance media. The lipid content assay and the relative expressions of PPAR-γ, ADR1, GLUT1 and GLUT4 genes showed no significant variation with and without the gold nanoparticles treatment. Differential phosphorylation levels of 43 kinases phosphorylation sites were evaluated using the human phospho-kinase array to assess the effect of the gold nanoparticles on the signaling pathways during the differentiation. No kinase phosphorylation site was differentially phosphorylated with two or more folds after the nanoparticles treatment after the first day as well as at the end of MG-63 cells differentiation. The gold-aryl nanoparticles do not affect MG-63 cells differentiation into adipocytes neither do they affect any key signaling pathway. These properties make these gold nanoparticles suitable for future drug delivery and medical applications.