Biodegradable PEG-PCL Nanoparticles for Co-delivery of MUC1 Inhibitor and Doxorubicin for the Confinement of Triple-Negative Breast Cancer

Combating triple-negative breast cancer (TNBC) is still a problem, despite the development of numerous drug delivery approaches. Mucin1 (MUC1), a glycoprotein linked to chemo-resistance and progressive malignancy, is unregulated in TNBC. GO-201, a MUC1 peptide inhibitor that impairs MUC1 activity, promotes necrotic cell death by binding to the MUC1-C unit. The current study deals with the synthesis and development of a novel nano-formulation (DM-PEG-PCL NPs) comprising of polyethylene glycol-polycaprolactone (PEG-PCL) polymer loaded with MUC1 inhibitor and an effective anticancer drug, doxorubicin (DOX). The DOX and MUC1 loaded nanoparticles were fully characterized, and their different physicochemical properties, viz. size, shape, surface charge, entrapment efficiencies, release behavior, etc., were determined. With IC(50) values of 5.8 and 2.4 nm on breast cancer cell lines, accordingly, and a combination index (CI) of < 1.0, DM-PEG-PCL NPs displayed enhanced toxicity towards breast cancer cells (MCF-7 and MDA-MB-231) than DOX-PEG-PCL and MUC1i-PEG-PCL nanoparticles. Fluorescence microscopy analysis revealed DOX localization in the nucleus and MUC1 inhibitor in the mitochondria. Further, DM-PEG-PCL NPs treated breast cancer cells showed increased mitochondrial damage with enhancement in caspase-3 expression and reduction in Bcl-2 expression.In vivo evaluation using Ehrlich Ascites Carcinoma bearing mice explicitly stated that DM-PEG-PCL NPs therapy minimized tumor growth relative to control treatment. Further, acute toxicity studies did not reveal any adverse effects on organs and their functions, as no mortalities were observed. The current research reports for the first time the synergistic approach of combination entrapment of a clinical chemotherapeutic (DOX) and an anticancer peptide (MUC1 inhibitor) encased in a diblock PEG-PCL copolymer. Incorporating both DOX and MUC1 inhibitors in PEG-PCL NPs in the designed nanoformulation has provided chances and insights for treating triple-negative breast tumors. Our controlled delivery technology is biodegradable, non-toxic, and anti-multidrug-resistant. In addition, this tailored smart nanoformulation has been particularly effective in the therapy of triple-negative breast cancer. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10924-022-02654-4

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Not AvailableSeafood is not considered the natural habitat of Salmonella except the river fish, but still, the incidence of Salmonella in seafood is in a steady rise. By extending our understanding of Salmonella growth dynamics and pathogenomics in seafood, we may able to improve seafood safety and offer better strategies to protect the public health. The current study was thus aimed to assess the growth and multiplication of non-typhoidal and typhoidal Salmonella serovars on seafood and further sought to evaluate their virulence and stress genes expression while in contact with seafood at varying temperature exposure. Salmonella enterica Weltevreden and Salmonella enterica Typhi were left to grow on fish fillets at −20, 4, room temperature (RT) and 45 °C for a period of one week. Total RNA from both Salmonella serovars were extracted and qRT-PCR based relative gene expression approach was used to detect the expression of rpoE, invA, stn and fimA genes at four different temperature conditions studied on incubation days 0, 1, 3, 5 and 7. Salmonella Weltevreden growth on seafood was increased ~4 log10 at RT and 45 °C, nevertheless, nearly 2 and >4 log 10 reduction was observed in cell count stored at 4 and −20 °C on seafood, respectively. Growth pattern of Salmonella Typhi in seafood has shown identical pattern at RT and 45 °C, however, growth was sharply reduced at 4 and −20 °C as compared to the Salmonella Weltevreden. Total RNA of Salmonella Weltevreden was in the range from 1.3 to 17.6 μg/μl and maximum concentration was obtained at 45 °C on day 3. Similarly, RNA concentration of Salmonella Typhi was ranged from 1.2 to 11.8 μg/μl and maximum concentration was obtained at 45 °C on day 3. The study highlighted that expression of invA and stn genes of Salmonella Weltevreden was >8-fold upregulated at RT, whereas, fimA gene was increasingly down regulated at room temperature. Storage of Salmonella Weltevreden at 45 °C on seafood resulted in an increased expression (>13 -fold) of stn genes on day 1 followed by down regulation on days 3, 5, and 7. Nevertheless, other genes i.e. fimA, invA and rpo remained down regulated throughout the storage period. More intense upregulation was observed for invA and stn genes of Salmonella Typhi at RT and 45 °C. Further, incubating Salmonella Weltevreden at 4 °C resulted in down regulation in the expression of rpoE, invA and stn genes. Regarding Salmonella Typhi, fimA and stn genes were upregulated on day one, in addition, an increased expression of fimA was noted on day 3. At −20 °C, there was no obvious expression of target genes of Salmonella Weltevreden and Salmonella Typhi when stored along with seafood. Here we demonstrate that nutritional constituents and water content available in seafood has become useful growth ingredients for the proliferation of Salmonella in a temperature dependent manner. Although, it was absence of serovar specific growth pattern of non-typhoidal and typhoidal Salmonella in seafood, there was observation of diverse expression profile of stress and virulent genes in non-typhoidal and typhoidal Salmonella serovars. In presence of seafood, the induced expression of Salmonella virulent genes at ambient temperature is most likely to be impacted by increased risk of seafood borne illness associated with Salmonella.Not Availabl

Circulatory extracellular vesicle derived miR-195-5p promotes cellular apoptosis and suppresses cell proliferation in the buffalo endometrial primary cell culture

Abstract In pregnant animals, communication between the mother and conceptus occurs via extracellular vesicles (EVs) that carry several biomolecules such as nucleic acids (miRNAs, mRNAs), proteins, and lipids. At the time of implantation, the endometrium undergoes several morphological and physiological changes, such as angiogenesis, apoptosis, and cell proliferation regulation at the implantation site, to attain a receptive state. This study was conducted to detect pregnancy-specific miRNAs derived from extracellular vesicles in the systemic circulation of Bubalus bubalis (water buffalo) and to assess their functional significance in the modulation of endometrial primary cells. The extracellular vesicles were isolated from the blood plasma using a precipitation-based method and further characterized by various methods such as Differential light scattering, Nanoparticle tracking assay, Western blot, and transmission electron microscopy. The relative expression of the selected extracellular vesicles associated miRNAs (EV-miRNA) at different intervals (days 15, 19, 25, and 30) post artificial insemination (AI) was analyzed using RT-qPCR, and expression of miR-195-5p was found to be significantly higher (P < 0.01) in pregnant animals on day 19 post AI (implantation window) as compared to day 15 post AI. The elevated expression might indicate the involvement of this miRNA in the maternal-conceptus cross-talk occurring during the implantation period. The KEGG pathway enrichment and Gene Ontology analyses of the miR-195-5p target genes revealed that these were mostly involved in the PI3-Akt, MAPK, cell cycle, ubiquitin-mediated proteolysis, and mTOR signaling pathways, which are related to the regulation of cell proliferation. Transfecting the in vitro cultured cells with miR-195-5p mimic significantly suppressed (P < 0.05) the expression of its target genes such as YWHAQ, CDC27, AKT-3, FGF-7, MAPK8, SGK1, VEGFA, CACAND1, CUL2, MKNK1, and CACAN2D1. Furthermore, the downregulation of the miR-195-5p target genes was positively correlated with a significant increase in the apoptotic rate and a decrease in the proliferation. In conclusion, the current findings provide vital information on the presence of EV miR-195-5p in maternal circulation during the implantation window indicating its important role in the modulation of buffalo endometrium epithelial cells via promoting cell death. Altogether, the milieu of miR-195-5p may serve as a novel and potential molecular factor facilitating the implantation of the early embryo during the establishment of pregnancy in buffaloes. Thus, miR-195-5p may be identified as a unique circulatory EV biomarker related to establishing pregnancy in buffaloes as early as day 19 post-AI

Production and Profitability of Hybrid Rice Is Influenced by Different Nutrient Management Practices

The government of Nepal has recommended blanket fertilizer application for rice cultivation, which results in lower nutrient use efficiency (NUE) particularly under rainfed conditions. With the aim of finding an appropriate nutrient management practices concerning rice production and profitability, a field experiment was conducted during rainy season of 2017 and 2018 at Kavrepalanchowk and Dang district of Nepal. Altogether, five treatments comprising various nutrient management practices viz. Nutrient Expert Model (NE), use of Leaf Color Chart (LCC), Government Recommended Fertilizer Dose (GON), Farm Yard Manure (FYM), and Farmers’ Field Practice (FFP), were laid out in RCBD with four replications in farmers’ fields. The analysis of variance showed significant difference between treatments for test weight and grain yield in Kavrepalanchowk whereas all traits except number of effective tillers were significant in Dang. The significantly higher grain yield and harvest index were obtained in NE, followed by LCC; and the overall straw yield was highest in LCC, followed by NE in both the locations. Also, yield gap analysis suggested the NE had 44.44% and 23.97% increase in yield as compared to FPP in Kavrepalanchowk and Dang, respectively. The combined analysis with Best Linear Unbiased Estimator revealed the interaction of nutrient management and location significantly effects the straw yield and harvest index across both the locations. The estimated mean straw yield and harvest index were 10.93 t/ha and 34.98%, respectively. Both correlation study and biplot of principal component analysis signaled grain yield had positive correlation with all other traits. Furthermore, the net revenue was maximum for NE, followed by LCC in both the locations. The benefit: cost ratio was highest for NE which was 1.55 in Kavrepalanchowk and 2.61 in Dang. On the basis of these findings, NE and LCC can be effectively used as nutrient management practice by the farmers to obtain maximum production and profitability in Rice

Production and Profitability of Hybrid Rice Is Influenced by Different Nutrient Management Practices

The government of Nepal has recommended blanket fertilizer application for rice cultivation, which results in lower nutrient use efficiency (NUE) particularly under rainfed conditions. With the aim of finding an appropriate nutrient management practices concerning rice production and profitability, a field experiment was conducted during rainy season of 2017 and 2018 at Kavrepalanchowk and Dang district of Nepal. Altogether, five treatments comprising various nutrient management practices viz. Nutrient Expert Model (NE), use of Leaf Color Chart (LCC), Government Recommended Fertilizer Dose (GON), Farm Yard Manure (FYM), and Farmers&rsquo; Field Practice (FFP), were laid out in RCBD with four replications in farmers&rsquo; fields. The analysis of variance showed significant difference between treatments for test weight and grain yield in Kavrepalanchowk whereas all traits except number of effective tillers were significant in Dang. The significantly higher grain yield and harvest index were obtained in NE, followed by LCC; and the overall straw yield was highest in LCC, followed by NE in both the locations. Also, yield gap analysis suggested the NE had 44.44% and 23.97% increase in yield as compared to FPP in Kavrepalanchowk and Dang, respectively. The combined analysis with Best Linear Unbiased Estimator revealed the interaction of nutrient management and location significantly effects the straw yield and harvest index across both the locations. The estimated mean straw yield and harvest index were 10.93 t/ha and 34.98%, respectively. Both correlation study and biplot of principal component analysis signaled grain yield had positive correlation with all other traits. Furthermore, the net revenue was maximum for NE, followed by LCC in both the locations. The benefit: cost ratio was highest for NE which was 1.55 in Kavrepalanchowk and 2.61 in Dang. On the basis of these findings, NE and LCC can be effectively used as nutrient management practice by the farmers to obtain maximum production and profitability in Rice