Ethanolic extract of Euphorbia royleana Boiss. reduces metastasis of breast cancer cells and inhibits tumor progression in vivo.

Metastasis is the most devastating attribute of breast cancer (BC) that leads to high mortality. It is a complex process of tumor cell migration, invasion, and angiogenesis. In this study, we evaluated the effect of ERA on BC metastasis and BC progression in vivo. The transwell invasion/migration and wound healing assays showed that ERA treatment significantly reduced the invasion and migration of BC cell lines. The expression of mesenchymal (E-cadherin and N-cadherin), matrix metalloproteinases (MMP2, MMP9), and stemness markers (Oct3) were down-regulated by ERA. Furthermore, ERA down-regulated angiogenic chemokines (CXCL1/2/3, CXCL5, and CXCL12) expression in the highly metastatic MDA-MB-231 cell line. The clonogenic survival of BC cells was also reduced by ERA treatment. Strikingly, ERA prevented DMBA-induced tumor growth in Swiss albino mice as depicted by a high animal survival rate (84%) in the ERA group and histopathological analysis. Conclusively, this study revealed that ERA possesses anti-metastatic potential and also reduces the growth of BC in vivo. Moreover, the GC-MS data revealed the presence of biologically active compounds (Lupeol, Phytol, phytosterol) and some rare (9, 19-Cyclolanost) phyto metabolites in ERA extract. However, further studies are suggestive to identify and isolate the therapeutic agents from ERA to combat BC and metastasis.

A Comparative Analysis on the Innate Immune Responses of Cirrhinus mrigala Challenged with Pseudomonas aeruginosa and Fusarium oxysporum.

Microbes are the most significant ubiquitous pathogens that cause serious infections in freshwater fish, leading to tremendous economic losses. The present study was designed to investigate the extent of changes in cytokine expression, hemato-biochemical parameters, and tissue histology of Cirrhinus mrigala (C. mrigala) challenged with Pseudomonas aeruginosa (P. aeruginosa) and Fusarium oxysporum (F. oxysporum). Fish were divided into three major groups: control, P. aeruginosa-challenged, and F. oxysporum-challenged. The infection in both challenge assays was allowed to progress until 7 days post infection. Upregulated expression of TNF-α and IL-1ß was found in blood, gills, livers, and kidneys of the challenged fish. Significant differences were noted in hematological parameters of challenged fish. Alanine aminotransferase, aspartate aminotransferase, and alkaline aminotransferase levels also showed significant differences in infected and control groups. An increase in serum albumin and globulin and a decrease in total protein were noted in infected groups as compared to the control group. Severe histological alterations were noted in gill, liver, and kidney tissues of the infected groups as compared to control. The order of histological alteration index for P. aeruginosa challenge was liver > kidney > gills, and for F. oxysporum challenge it was kidney > liver > gills. These changes in fish infected by P. aeruginosa and F. oxysporum can be used as an effective and subtle index to monitor the physiological and pathological conditions of fish.

Autochthonous strains of Trichoderma isolated from tannery solid waste improve phytoextraction potential of heavy metals by sunflower.

This research work was aimed at isolating and demonstrating the significant potential of autochthonous fungi for phytoextraction of hazardous metals in metal polluted soil using Helianthus annuus. Four multi-metal resistant strains of Trichoderma were selected from a total of 21 strains isolated from tannery polluted soil and tannery solid waste. Autochthonous Trichoderma strains were used singly and in the form of consortium (TC). Sunflower was grown in pots for 90 days having eight different amendments of tannery polluted soil with and without Trichoderma inoculation. Growth and biochemical attributes of the plants were observed along with metal content extract by different plant parts. The results revealed that TC enhanced shoot length, shoot dry weight, and metal uptake as compared to single specie inoculation. Similarly, BCF (72.8-118.23%) and TF were significantly pronounced in shoots of H. annuus grown with TC at 40% amended soil. The biochemical analysis of the plants showed that Trichoderma strains boosted the enzymatic (catalase, peroxidase, and superoxide dismutase) antioxidants in the plants. The use of indigenous fungi with metal accumulating plants like sunflower can help to alleviate metal contamination from industrial sites and can make the soil cultivable for energy crops.

The genus Trichoderma is among the most common cosmopolitan soil fungi that enhance phytoextraction capability of plants. Hence, the isolation and identification of diversified and potent Trichoderma strains from contaminated environments is the need of the hour for broad spectrum applications in bioremediation. In the present study, contaminated soil mycoflora was explored and multi-metal resistant strains of Trichoderma were isolated. Their application in myco-assisted phytoextraction with Helianthus annuus was assessed to analyze their impact on the metal removal efficacy and enhancing growth in highly contaminated soil.

An Insight into the Role of E2F1 in Breast Cancer Progression, Drug Resistance, and Metastasis.

AIMS: This study aimed to investigate the role of E2F1 in breast cancer biology. BACKGROUND: Expression of E2F1, a transcription factor of many oncogenes and tumor suppressor genes, is lowered in several malignancies, including breast carcinoma. OBJECTIVES: In the present study, we analyzed the status of E2F1 expression in association with diverse attributes of breast malignancy and its impact on cancer progression. METHODS: For this purpose, we used various freely available online applications for gene enrichment, expression, and methylation analysis to extract mutation-based E2F1 map, to measure E2F1 drug sensitivity, and to determine E2F1 association with DNA damage response proteins. RESULTS: Results revealed tissue-specific regulatory behavior of E2F1. Moreover, the key role of E2F1 in the promotion of metastasis, stem cell-mediated carcinogenesis, estrogen-mediated cell proliferation, and cellular defense system, has therefore highlighted it as a metaplastic marker and hot member of key resistome pathways. CONCLUSION: The information thus generated can be employed for future implications in devising rational therapeutic strategies. Moreover, this study has provided a more detailed insight into the diagnostic and prognostic potential of E2F1.

Equilibrium kinetics and thermodynamic studies on biosorption of heavy metals by metal-resistant strains of Trichoderma isolated from tannery solid waste.

This study was aimed at finding the metal sorption potential of six indigenous Trichoderma strains by using batch experiments for Cd (II), Cr (VI), Cu (II), and Pb (II). Trichoderma atrobrunneum showed maximum metal biosorption potential at 800 mg L-1 of initial concentration. Two adsorption isotherm models, (1) Langmuir (2) Freundlich models, were employed on the biosorption data obtained at various initial metal concentrations (10 mg L-1-200 mg L-1) and pseudo-first (PSI) and pseudo-second (PSII) order equilibrium kinetic models were subjected to data of agitation time (3-7 days). A maximum correlation coefficient value (R2) of ≤ 1 was observed for the Langmuir and PSII model. Results revealed that pH 6-7 was the best for metal sorption, while metal removal efficiency was increased by increasing temperature (298 K, 303 K, 308 K, 313 K). The results of thermodynamic study parameters (∆G°, ∆H°, ∆S°) indicated that heavy metal biosorption by Trichoderma strains was an endothermic, spontaneous, and feasible process. Moreover, surface characterization analysis through SEM, BET, FTIR, and XRD showed that T. atrobrunneum and Trichoderma sp. could adsorb more metal ions when grown in high metal concentrations. The results indicate that living biomass of T. atrobrunneum and Trichoderma sp. is an effective multi-metal biosorbent that can be used for efficacious bioremediation of bio-treatment of heavy metal polluted wastewater.

GABRG2 C588T Polymorphism Is Associated with Idiopathic Generalized Epilepsy but Not with Antiepileptic Drug Resistance in Pakistani Cohort.

Idiopathic generalized epilepsy (IGE) is the most prevalent type of epilepsy with genetic origin. Mutations in ion channel genes have been identified as a common cause of IGE. Several studies have reported various epilepsy risk variants of GABRG2 (gamma-aminobutyric acid type A receptor subunit gamma2 subunit) gene in different ethnic groups, but the results are inconsistent. The purpose of this case-control research is to determine if GABRG2 polymorphisms contribute to IGE susceptibility and antiepileptic drug resistance in Pakistani population. For this purpose, we genotyped exon2, exon5 (C540T and C588T), exon7 (T813C), exon8 (K289M), and exon9 of GABRG2 gene by restriction fragment length polymorphism and Sanger's sequencing in 87 drug-responsive idiopathic generalized epilepsy patients, 55 drug-resistant epilepsy patients, and 83 healthy controls. Restriction fragment length polymorphism (RFLP) and sequencing results indicated only C588T polymorphism in the studied subjects. The comparison of genotypic and allelic frequencies showed significant differences between IGE patients and control groups (P = 0.008 and odds ratio = 4.2) and nonsignificant association of C588T polymorphism in antiseizure medication-resistant patients (P = 0.9). Our findings showed that C588T polymorphism of GABRG2 is a risk variant for IGE in Pakistani population. Further studies are required to validate the results.

Role of Solvent Used in Development of Graphene Oxide Coating on AZ31B Magnesium Alloy: Corrosion Behavior and Biocompatibility Analysis.

Clinical applications of bio-absorbable magnesium (Mg) and its alloys can be enhanced by increasing their corrosion resistance, using surface modification and functionality. In this study, we synthesized graphene oxide (GO) through improved Hummers' method and deposited it on biodegradable AZ31B Mg alloy for further characterization. Different suspensions of GO were prepared in various solvents, like deionized water, ethanol, and acetone by ultra-sonication. Electrophoretic deposition (EPD) was used to develop GO coatings on AZ31B Mg using different GO suspensions. Effect of various solvents on corrosion behavior, as well as in vitro biocompatibility, was studied. The optimized EPD parameters were 3 volts and 90 s for coating. Different characterization techniques were used to study GO and prepared coatings. Atomic force microscopy found that the average thickness of GO was ~1 nm. Electrochemical behavior of coatings was studied through electrochemical impedance spectroscopy (EIS) and Tafel analysis in Ringer's lactate solution. Tafel analysis revealed that GO coatings deposited by GO water suspension increased corrosion protection efficiency of AZ31B Mg alloy by ~94%. After 72 h incubation in MC3T3-E1 osteoblast cells extract, in vitro analysis was performed to determine the cell viability and biocompatibility of the GO- coated and bare Mg samples. GO coatings deposited by GO water suspension demonstrated ~2× cell viability, as well as nontoxicity and better biocompatibility compared to the bare and other GO-coated Mg samples.

Citrullus colocynthis-Mediated Green Synthesis of Silver Nanoparticles and Their Antiproliferative Action against Breast Cancer Cells and Bactericidal Roles against Human Pathogens.

The present study investigated the biomedical potential of eco-friendly Citrullus colocynthis-mediated silver nanoparticles (Cc-AgNPs). The antibacterial efficacy of Cc-AgNPs was evaluated against two multidrug-resistant pathogenic bacterial strains, Escherichia coli and Pseudomonas aeruginosa. The antiproliferative and antilipidemic performance of the prepared particles was determined against the MCF7 cell line, a breast cancer cell line. The in vitro antibacterial assay revealed that Cc-AgNPs induced dose-dependent bactericidal activity, as a considerable increase in the zone of inhibition (ZOI) was noted at higher concentrations. Reduced proliferation, migration, spheroid size, and colony formation exhibited the substantial antiproliferative potential of Cc-AgNPs against MCF7 cells. Significant alterations in the expression of cell surface markers, apoptosis, and cell proliferation genes further confirmed the antiproliferative impact of Cc-AgNPs. Moreover, Cc-AgNPs exhibited antilipidemic activity by reducing cellular cholesterol and triglyceride levels and regulating key genes involved in lipogenesis. In conclusion, these results propose that Cc-AgNPs can be employed as a potent tool for future antibacterial and anticancer applications.

Inhibition of anti-inflammatory pathway through suppressors of cytokine signalling (Socs2/Socs3) in the initiation of hepatocellular carcinoma.

Hepatocellular carcinoma (HCC), a leading cause of cancer related deaths is predominantly driven by chronic inflammatory responses. Due to asymptomatic nature and lack of early patient biopsies, precise involvement of inflammation in hepatic injury initiation remains unidentified. Aim of the study was to elucidate the regulation patterns of inflammatory signalling from initiation of hepatic injury to development of HCC. HCC mice model was established using DEN followed by repeated doses of CCl4 and sacrificed at three different stages of disease comprising 7, 14 and 21 weeks. Serum biochemical tests, hepatic lipids quantification, histopathology and qPCR analyses were conducted to characterize the initiation and progression of liver injury and inflammatory signalling. Notably, at 7 weeks, we observed hepatocyte damage and periportal necrotic bodies coupled with induction of Socs2/Socs3 and anti-inflammatory cytokine Il-10. At 14 weeks, mice liver showed advancement of liver injury with micro-vesicular steatosis and moderate collagen deposition around portal zone. With progression of injury, the expression of Socs3 was declined with further reduction of Il-10 and Tgf-ß indicating the disturbance of anti-inflammatory mechanism. In contrast, pro-inflammatory cytokines Il1-ß, Il6 and Tnf-α were upregulated contributing inflammation. Subsequently, at 21 weeks severe liver damage was estimated as characterized by macro-vesicular steatosis, perisinusoidal collagen bridging, immune cell recruitment and significant upregulation of Col-1α and α-Sma. In parallel, there was significant upregulation of pro/anti-inflammatory cytokines highlighting the commencement of chronic inflammation. Findings of the study suggest that differential regulation of cytokine suppressors and inflammatory cytokines might play role in the initiation and progression of hepatic injury leading towards HCC.

A comparison of total RNA extraction methods for RT-PCR based differential expression of genes from Trichoderma atrobrunneum.

The genus Trichoderma is ubiquitous in various niches and is currently used for biocontrol, biofertilizer, enzyme production and bioremediation. However, molecular mechanisms underlying its diverse biological functions are yet not fully elucidated. Extraction of high-quality RNA for downstream applications such as quantitative polymerase chain reaction is a prerequisite. The current study aims to optimize a total RNA extraction protocol for high-quality and quantity RNA from Trichoderma atrobrunneum. Seven RNA extraction protocols including Trizol, RiboEx PureLink RNA mini kit, high salt CTAB (cetyltrimethylammonium bromide), modified high salt CTAB, SDS (sodium dodecyl sulfate) and CTAB-PVP (polyvinylpyrrolidone) were performed separately, to extract RNA. Quality and quantity of extracted RNA samples were further analyzed by Nanodrop spectrophotometry, agarose gel electrophoresis and RT-PCR analysis. The results of quantitative and qualitative analysis of RNA samples showed that more intact, high-quality RNA was extracted using the modified high salt CTAB as compared to other methods. The RT-PCR results for the amplification of the genes encoding ß-tubulin and Ubiquitin carrier protein also showed lowest threshold cycle (Ct) and coefficient of variation (CV) for RNA samples extracted with the modified high salt CTAB method as compared to RNA samples extracted with other protocols. Therefore, it is proposed that the modified high salt CTAB protocol is an excellent method to obtain high-quality RNA with good yield from T. atrobrunneum for its downstream applications. Moreover, the optimized protocol is very economical and can be used to extract total RNA from a large number of samples.

Anti-diabetic and anti-lipidepmic effect of Albizia lebbeck seeds against HepG2 cells.

Albizia lebbeck has been a medicinally important plant for its pharmacological potential. This study aims to determine the in vitro antioxidant, anti-diabetic and anti-lipidemic potential of A. lebbeck seeds. The seed extracts were prepared in petroleum ether, chloroform and methanol. Crude methanolic extract (ME ext) was subjected further to sequential fractionation in increasing polarity based solvents. Extracts and fractions were analyzed for their antioxidant, anti-diabetic and anti-lipidemic potentials using hepatic cell line, HepG2. Results showed that crude extracts of A. lebbeck seeds specifically, ME ext are rich in polyphenols and flavonoids. ME ext has also shown highly significant antioxidant and alpha-amylase inhibition potential compared to petroleum ether and chloroform extracts. In vitro assays using different fractions of methanolic extract further highlighted the ethyl acetate and chloroform fractions exhibiting significant antioxidant and anti-diabetic potentials. Alpha-amylase inhibition coupled with enhanced glucose uptake of cells treated with ME ext and ethyl acetate fraction emphasized on significant anti-diabetic potential of the plant. Expression alteration of genes and reduced level of cholesterol suggested the lipid synthesis mediated anti-diabetic activity of the plant. It is therefore, concluded that A. lebbeck seed has significant antioxidant, anti-diabetic and anti-lipidemic potentials.

Bisphenol S induced dysregulations in liver; iron regulatory genes and inflammatory mediators in male Wistar rats.

Bisphenol S (BPS), an analog of bisphenol A (BPA), has been frequently detected in consumer products, food wrappers, plastics, and thermal papers. Since the liver is a hub of metabolic and detoxification pathways, thus intimately related to BPS presence in the environment and body. The current study was designed to investigate the effects of BPS administration in an animal model. Twenty-five male Wistar rats weighing 175 ± 25 g were randomly divided into control and treated groups. The control group was further divided into group I (no treatment) and group II (corn oil), whereas the treatment group was divided into D-I (40 mg/kg/day), D-II (200 mg/kg/day), and D-III (400 mg/kg/day) groups, getting oral doses of BPS for 15 days. Data analysis showed a significant statistical increase in hepatic enzymes ALT (33.4%), AST (25.4%), and ALP (529.6%) in the D-III group along with the development of hypercholesterolemia and hypertriglyceridemia in all BPS groups. Aberrant mRNA expressions of some key hepatic iron regulatory genes and inflammatory mediators were evident through qRT-PCR. Bisphenol S caused congestion of central vein from mild to moderate in hepatic sections. In conclusion, our investigation insinuates BPS intoxication potential and therefore may not be a safe alternative to BPA.

Calreticulin Shortage Results in Disturbance of Calcium Storage, Mitochondrial Disease, and Kidney Injury.

Renal Ca2+ reabsorption plays a central role in the fine-tuning of whole-body Ca2+ homeostasis. Here, we identified calreticulin (Calr) as a missing link in Ca2+ handling in the kidney and showed that a shortage of Calr results in mitochondrial disease and kidney pathogenesis. We demonstrated that Calr+/- mice displayed a chronic physiological low level of Calr and that this was associated with progressive renal injury manifested in glomerulosclerosis and tubulointerstitial damage. We found that Calr+/- kidney cells suffer from a disturbance in functionally active calcium stores and decrease in Ca2+ storage capacity. Consequently, the kidney cells displayed an abnormal activation of Ca2+ signaling and NF-κB pathways, resulting in inflammation and wide progressive kidney injury. Interestingly, the disturbance in the Ca2+ homeostasis and signaling in Calr+/- kidney mice cells triggered severe mitochondrial disease and aberrant mitophagy, resulting in a high level of oxidative stress and energy shortage. These findings provide novel mechanistic insight into the role of Calr in kidney calcium handling, function, and pathogenesis.

Ethanolic extracts of Pakistani euphorbiaceous plants induce apoptosis in breast cancer cells through induction of DNA damage and caspase-dependent pathway.

BACKGROUND: Numerous plants of Euphorbiaceae, thespurgefamily are traditionally used for the treatment of different diseases and recent studies also reported anti-oxidant, anti-inflammatory, and anti-tumor activities of these plants. However, the medicinal potential of several indigenous euphorbiaceous plants of Pakistan is not described yet. Therefore, we intended to evaluate the in vitro anti-breast cancer potential of 10 euphorbiaceous plants of Pakistan. METHODS: Cytotoxic screening of ethanolic extracts of selected plants was performed by MTT assay. The qualitative phytochemical analysis was performed to find the major groups of chemicals responsible for cytotoxic activity. To determine the genotoxic effect of plant extracts, microscopic analysis was carried out. Flow cytometry and fluorescent microscopic analysis were done to detect apoptosis. To find out the expression analysis of cell cycle and cell death regulatory genes, quantitative real-time polymerase reaction (qRT-PCR) was performed. RESULTS: Among the 10 tested plants, ethanolic extracts of Croton tiglium (CTL) and Euphorbia royleana (ERA) were found to possess the highest anti-proliferative activity against breast cancer cells (MDA-MB-231, MCF-7), with IC50 values 100 and 80 µg/mL respectively. The phytochemical analysis confirmed the presence of phenols, flavonoids, and steroids in both plant extracts, whereas, glycosides and saponins were found only in CTL and ERA, respectively. The cellular aberrations and nuclear morphologies with a distinct DNA laddering pattern substantiated the genotoxic effects. Furthermore, our data showed that CTL and ERA induce cell cycle arrest at the G1/S phase by down-regulating the CDK4 and Cyclin D1 expression followed by caspase-dependent induction of apoptosis in both MCF-7 and MDA-MB-231 cells. However, based on the activation of initiator and executioner caspases, two distinct types of apoptotic pathways are proposed for these plants. The CTL prompted extrinsic while ERA triggered the intrinsic pathways of apoptosis. CONCLUSION: Our data demonstrate the strong anti-proliferative and caspase-dependent apoptotic potential of CTL and ERA against breast cancer cells. Further studies are suggested to find clinical implications of these plants in breast cancer therapeutic.

In vitro antioxidant potential and antiproliferative activity of Aesculus indica seed extract against hepatocellular carcinoma cell line.

Therapeutic secondary metabolites have gained immense attention in recent years due to their effective medicinal properties. Aesculus indica is a medicinally important plant being traditionally used for various ailments. The present study aimed to determine the antioxidant and antiproliferative activities of seeds of A. indica. The crude methanolic seed extract was prepared and subjected to sequential fractionation in increasing polarity. The extract and its fractions were investigated for antioxidant activities using various in vitro assays. Further, the extract along with its potential antioxidant fractions were analyzed for their cytotoxic activity against HepG2, human hepatocyte carcinoma cells through bioassays. The results showed highly significant antioxidant potential of methanolic extract of A. indica seeds and two of its fractions prepared with chloroform and ethyl acetate. The studies on hepatocyte carcinoma cells further revealed that the extract and two of its potential antioxidant fractions significantly induced cytotoxicity and inhibited migration, proliferation, clonogenicity and 3D growth of HepG2 cells. It is therefore, concluded that A. indica possess significant antioxidant and cytotoxic potential against HepG2 cells and with further research can be proposed for therapeutic interventions.

Calreticulin Deficiency Disturbs Ribosome Biogenesis and Results in Retardation in Embryonic Kidney Development.

Nephrogenesis is driven by complex signaling pathways that control cell growth and differentiation. The endoplasmic reticulum chaperone calreticulin (Calr) is well known for its function in calcium storage and in the folding of glycoproteins. Its role in kidney development is still not understood. We provide evidence for a pivotal role of Calr in nephrogenesis in this investigation. We show that Calr deficiency results in the disrupted formation of an intact nephrogenic zone and in retardation of nephrogenesis, as evidenced by the disturbance in the formation of comma-shaped and s-shaped bodies. Using proteomics and transcriptomics approaches, we demonstrated that in addition to an alteration in Wnt-signaling key proteins, embryonic kidneys from Calr-/- showed an overall impairment in expression of ribosomal proteins which reveals disturbances in protein synthesis and nephrogenesis. CRISPR/cas9 mediated knockout confirmed that Calr deficiency is associated with a deficiency of several ribosomal proteins and key proteins in ribosome biogenesis. Our data highlights a direct link between Calr expression and the ribosome biogenesis.

Methanolic extract of Citrullus colocynthis suppresses growth and proliferation of breast cancer cells through regulation of cell cycle.

Breast cancer is a major cause of cancer related deaths in women worldwide. Available treatments pose serious limitations such as systemic toxicity, metastasis, tumor recurrence, off-target effects, and drug resistance. In recent years, phytochemicals such as secondary metabolites due to their effective anticancer potential at very low concentration have gained attention. Aim of the study was to evaluate anticancer potential of Citrullus colocynthis and its possible molecular targets on MCF-7, a human breast cancer cell line. Methanolic extract of leaves was prepared and fractionated by solvents (n-hexane, chloroform, ethyl acetate and n-butanol) with increasing polarity. Bioassays and gene expression regulation was conducted to evaluate the anticancer activity, proliferation rate and cell cycle regulation of breast cancer cells treated with extract and its fractions, separately. Results showed a significant anticancer activity of methanolic extract of C. colocynthis and two of its fractions prepared with chloroform and ethyl acetate. Bioassays depicted significant decrease in proliferation and growth potential along with cell cycle arrest of treated cells compared to control untreated cells. Expression regulation of genes further confirmed the cell cycle arrest through significant upregulation of cyclin-CDK inhibitors (p21 and p27) and cell cycle checkpoint regulators (HUS1, RAD1, ATM) followed by downregulation of downstream cell cycle progression genes (Cyclin A, Cyclin E, CDK2). It is concluded that C. colocynthis arrests cell cycle in human breast cancer cells through expression regulation of cyclin-CDK inhibitors and with further research can be proposed for therapeutic interventions.

High-Fat Diet Induced Hedgehog Signaling Modifications during Chronic Kidney Damage.

Excessive consumption of dietary fats leads to the deposition of unnecessary metabolites and multiple organ damage. Lipids, important key regulators of Hedgehog signaling, are involved in triggering fibrotic chronic kidney disease. The present study encompasses the assessment of renal morphofunctional modifications and alteration of lipid metabolism influencing the changes in gene expression of hedgehog signaling pathway genes. Fifteen male Rattus norvegicus of 200 ± 25 grams weight were equally divided into three groups: control (standard rat chow), D-1 (unsaturated high-fat diet) and D-2 (saturated high-fat diet). Animals were provided with respective diets and were followed for 16 weeks. Both HFD-fed groups did not show overall body weight gain as compared to the control. While significant downregulation of hedgehog pathway genes was found in fatty diet groups. In comparison with the control group, Shh, Gli1, Gli2, and Gli3 were downregulated after the consumption of both unsaturated and saturated fatty diets. Ihh and Smo exhibit a similar downregulation in the D-1 group, but an upregulation was detected in the D-2 group. D-2 group also had an increased serum urea concentration as compared to the control (P = 0.0023). Furthermore, renal histopathology revealed tubular necrosis, glomerular edema, glomerular shrinkage, and hypocellularity. Collagen deposition in both HFD groups marks the extent of fibrosis summary figure. Extravagant intake of dietary fats impaired normal kidney functioning and morphofunctionally anomalous kidney triggers on Hh signaling in adult rats. These anomalies can be linked to an escalated risk of chronic kidney disease in adults strongly recommending the reduced uptake of fatty diets to prevent impaired metabolism and renal lipotoxicity.

Citrullus colocynthis regulates de novo lipid biosynthesis in human breast cancer cells.

BACKGROUND: Reprogrammed energy metabolism is considered a hallmark of cancer and is proposed as an important target for therapy. Uncontrolled and infinite cell proliferation needs efficient energy sources. To meet the demands of cancer cells lipid metabolism is activated. Citrullus colocynthis is a traditional medicinal plant known for its anticancer and hypolipidemic effects. AIMS: Aim of the current study was to assess the effect of C. colocynthis leaves on regulation of lipid metabolism in MCF-7, a human breast cancer cell line. METHODS: Methanolic extract of leaves and its fractions in increasing polarity-based solvents (n-hexane, chloroform, ethyl acetate and n-butanol) were prepared and analyzed for the presence of secondary metabolites in each fraction. Bioassays and apoptosis genes expression analysis was conducted to evaluate the anticancer and cytotoxic effect of breast cancer cells treated with extract and its fractions, separately. Lipid quantification and gene expression regulation of genes involve in lipid metabolism was performed to evaluate regulation of lipid metabolism. RESULTS: Results showed a significant anticancer activity of methanolic extract of C. colocynthis and two of its fractions prepared with chloroform and ethyl acetate. Quantification of lipids depicted significant increase in cholesterol and increase in triglycerides of treated cells compared to control untreated cells. Expression regulation of genes further confirmed the lipid regulation through significant down regulation of genes involve in lipid metabolism (FASN, HMGCLL1, ACSL5 and ELOVL2). CONCLUSION: The present study concludes that C. colocynthis holds strong anticancer potential through regulation of lipid metabolism and with further studies can be proposed for novel therapeutic approaches.

Next-generation sequencing to elucidate adaptive stress response and plantaricin genes among Lactobacillus plantarum strains.

Aim: The objective of this study was to identify the genes involved in plantaricin synthesis and adaptive stress response in four Lactobacillus plantarum strains (AS-6, AS-8, AS-9 and AS-10) and one Lactobacillus paraplantarum strain (AS-7) for their usage in medicine and industry. Materials & methods: Whole genomes of these strains were sequenced by a high-throughput sequencing technique known as next-generation sequencing via Ilumina MiSeq platform and the genes were identified by using various bioinformatics tools and software. Results: Plantaricin genes (plnD, plnE, plnF, plnG, plnI) and genes regulating response to temperature, pH, bile salt, osmotic and oxidative stress were identified in all strains. Conclusion: Lactobacilli could be an option to combat antimicrobial resistance and might replace harmful antibiotics in future.