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Heavy metal pollution is a significant environmental concern with detrimental effects on ecosystems and human health, and traditional remediation methods may be costly, energy-intensive, or have limited effectiveness. The current study aims were to investigate the impact of heavy metal toxicity in Eisenia fetida, the growth, reproductive outcomes, and their role in soil remediation. Various concentrations (ranging from 0 to 640 mg per kg of soil) of each heavy metal were incorporated into artificially prepared soil, and vermi-remediation was conducted over a period of 60 days. The study examined the effects of heavy metals on the growth and reproductive capabilities of E. fetida, as well as their impact on the organism through techniques such as FTIR, histology, and comet assay. Atomic absorption spectrometry demonstrated a significant (P < 0.000) reduction in heavy metal concentrations in the soil as a result of E. fetida activity. The order of heavy metal accumulation by E. fetida was found to be Cr > Cd > Pb. Histological analysis revealed a consistent decline in the organism's body condition with increasing concentrations of heavy metals. However, comet assay results indicated that the tested levels of heavy metals did not induce DNA damage in E. fetida. FTIR analysis revealed various functional group peaks, including N-H and O-H groups, CH2 asymmetric stretching, amide I and amide II, C-H bend, carboxylate group, C-H stretch, C-O stretching of sulfoxides, carbohydrates/polysaccharides, disulfide groups, and nitro compounds, with minor shifts indicating the binding or accumulation of heavy metals within E. fetida. Despite heavy metal exposure, no significant detrimental effects were observed, highlighting the potential of E. fetida for sustainable soil remediation. Vermi-remediation with E. fetida represents a novel, sustainable, and cutting-edge technology in environmental cleanup. This study found that E. fetida can serve as a natural and sustainable method for remediating heavy metal-contaminated soils, promising a healthier future for soil.
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Recuperação e Remediação Ambiental , Metais Pesados , Oligoquetos , Reprodução , Poluentes do Solo , Oligoquetos/efeitos dos fármacos , Metais Pesados/toxicidade , Animais , Poluentes do Solo/toxicidade , Reprodução/efeitos dos fármacos , Recuperação e Remediação Ambiental/métodos , Ensaio Cometa , Espectroscopia de Infravermelho com Transformada de Fourier , Dano ao DNA , Solo/químicaRESUMO
A tremendous increase in the green synthesis of metallic nanoparticles has been noticed in the last decades, which is due to their unique properties at the nano dimension. The present research work deals with synthesis mediated by the actinomycete Streptomyces tendae of silver nanoparticles (AgNPs), isolated from Little and Greater Rann of Kutch, India. The confirmation of the formation of AgNPs by the actinomycetes was carried out by using a UV-Vis spectrophotometer where an absorbance peak was obtained at 420 nm. The X-ray diffraction pattern demonstrated five characteristic diffraction peaks indexed at the lattice plane (111), (200), (231), (222), and (220). Fourier transform infrared showed typical bands at 531 to 1635, 2111, and 3328 cm-1. Scanning electron microscopy shows that the spherical-shaped AgNPs particles have diameters in the range of 40 to 90 nm. The particle size analysis displayed the mean particle size of AgNPs in aqueous medium, which was about 55 nm (±27 nm), bearing a negative charge on their surfaces. The potential of the S. tendae-mediated synthesized AgNPs was evaluated for their antimicrobial, anti-methicillin-resistant Staphylococcus aureus (MRSA), anti-biofilm, and anti-oxidant activity. The maximum inhibitory effect was observed against Pseudomonas aeruginosa at (8 µg/mL), followed by Escherichia coli and Aspergillus niger at (32 µg/mL), and against Candida albicans (64 µg/mL), whereas Bacillus subtilis (128 µg/mL) and Staphylococcus aureus (256 µg/mL) were much less sensitive to AgNPs. The biosynthesized AgNPs displayed activity against MRSA, and the free radical scavenging activity was observed with an increase in the dosage of AgNPs from 25 to 200 µg/mL. AgNPs in combination with ampicillin displayed inhibition of the development of biofilm in Pseudomonas aeruginosa and Streptococcus pneumoniae at 98% and 83%, respectively. AgNPs were also successfully coated on the surface of cotton to prepare antimicrobial surgical cotton, which demonstrated inhibitory action against Bacillus subtilis (15 mm) and Escherichia coli (12 mm). The present research integrates microbiology, nanotechnology, and biomedical science to formulate environmentally friendly antimicrobial materials using halotolerant actinomycetes, evolving green nanotechnology in the biomedical field. Moreover, this study broadens the understanding of halotolerant actinomycetes and their potential and opens possibilities for formulating new antimicrobial products and therapies.
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Bio-inspired zinc oxide nanoparticles are gaining immense interest due to their safety, low cost, biocompatibility, and broad biological properties. In recent years, much research has been focused on plant-based nanoparticles, mainly for their eco-friendly, facile, and non-toxic character. Hence, the current study emphasized a bottom-up synthesis of zinc oxide nanoparticles (ZnO NPs) from Psidium guajava aqueous leaf extract and evaluation of its biological properties. The structural characteristic features of biosynthesized ZnO NPs were confirmed using various analytical methods, such as UV-Vis spectroscopy, X-ray diffraction (XRD), energy-dispersive X-ray analysis (EDX), Fourier transform infrared spectroscopy (FT-IR), dynamic light scattering (DLS), Scanning electron microscopy (SEM) and high-resolution transmission electron microscopy (HR-TEM). The synthesized ZnO NPs exhibited a hydrodynamic shape with an average particle size of 11.6-80.2 nm. A significant antimicrobial efficiency with minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of 40 and 27 µg/ml for Enterococcus faecalis, followed by 30 and 40 µg/ml for Staphylococcus aureus, 20 and 30 µg/ml for Staphylococcus mutans, 30 µg/ml for Candida albicans was observed by ZnO NPs. Additionally, they showed significant breakdown of biofilms of Streptococcus mutans and Candida albicans indicating their future value in drug-resistance research. Furthermore, an excellent dose-dependent activity of antioxidant property was noticed with an IC50 of 9.89 µg/ml. The antiproliferative potential of the ZnO NPs was indicated by the viability of MDA MB 231 cells, which showed a drastic decrease in response to increased concentrations of biosynthesized ZnO NPs. Thus, the present results open up vistas to explore their pharmaceutical potential for the development of targeted anticancer drugs in the future.
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Propranolol (PRO) is a beta-blocker drug used for the treatment of anxiety, chest pain, migraine and tremors. The present study investigated whether sublethal concentrations of PRO have effects on the body condition, biochemistry, and hematology of Clarias gariepinus juveniles. The 96-h median lethal concentration (LC50) of the drug, which was established through an acute toxicity study, was 9.48 mg/L. Based on these values, the fish were exposed for 21 days to the control and sublethal concentrations of 1.90, 0.95, and 0.63 mg/L, which are equivalent to the 1/5th, 1/10th, and 1/20th of the LC50 of PRO, respectively. After 21 days of exposure, the fish were removed from the toxicant and kept in toxicant-free water for 7 days to recover. The standard length and body weight of each fish were measured after each exposure period. The condition factor (CF) and hepatosomatic index (HSI) were not significantly affected by the drug. The red blood cell (RBC) count, hemoglobin (Hb) count and packed cell volume (PCV) decreased from day 7 to 21 at the tested concentrations, while the white blood cell (WBC) count significantly increased. There were alterations in the mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), and mean corpuscular hemoglobin (MCH) in the exposed groups compared to those in the control group. When neutrophil counts increased, the lymphocyte counts decreased, but the monocyte, basophil cell and eosinophil cell counts were not affected. Among the liver enzymes, only aspartate aminotransferase was significantly stimulated in the groups that were exposed to the drug. The protein and glucose levels of fish exposed to the drug decreased. Most of the studied parameters returned to their original values after the 7-day recovery period. The information provided in the current study will be helpful in the monitoring of PRO contamination in aquatic environments.
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Background: The healing of burn wounds is a complicated physiological process that involves several stages, including haemostasis, inflammation, proliferation, and remodelling to rebuild the skin and subcutaneous tissue integrity. Recent advancements in nanomaterials, especially nanofibers, have opened a new way for efficient healing of wounds due to burning or other injuries. Methods: This study aims to develop and characterize collagen-decorated, bilayered electrospun nanofibrous mats composed of PVP and PVA loaded with Resveratrol (RSV) and Ampicillin (AMP) to accelerate burn wound healing and tissue repair. Results: Nanofibers with smooth surfaces and web-like structures with diameters ranging from 200 to 400 nm were successfully produced by electrospinning. These fibres exhibited excellent in vitro properties, including the ability to absorb wound exudates and undergo biodegradation over a two-week period. Additionally, these nanofibers demonstrated sustained and controlled release of encapsulated Resveratrol (RSV) and Ampicillin (AMP) through in vitro release studies. The zone of inhibition (ZOI) of PVP-PVA-RSV-AMP nanofibers against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) was found 31±0.09 mm and 12±0.03, respectively, which was significantly higher as compared to positive control. Similarly, the biofilm study confirmed the significant reduction in the formation of biofilms in nanofiber-treated group against both S. aureus and E. coli. X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR) analysis proved the encapsulation of RSV and AMP successfully into nanofibers and their compatibility. Haemolysis assay (%) showed no significant haemolysis (less than 5%) in nanofiber-treated groups, confirmed their cytocompatibility with red blood cells (RBCs). Cell viability assay and cell adhesion on HaCaT cells showed increased cell proliferation, indicating its biocompatibility as well as non-toxic properties. Results of the in-vivo experiments on a burn wound model demonstrated potential burn wound healing in rats confirmed by H&E-stained images and also improved the collagen synthesis in nanofibers-treated groups evidenced by Masson-trichrome staining. The ELISA assay clearly indicated the efficient downregulation of TNF-alpha and IL-6 inflammatory biomarkers after treatment with nanofibers on day 10. Conclusion: The RSV and AMP-loaded nanofiber mats, developed in this study, expedite burn wound healing through their multifaceted approach.
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Ampicilina , Queimaduras , Colágeno , Escherichia coli , Nanofibras , Álcool de Polivinil , Povidona , Resveratrol , Staphylococcus aureus , Cicatrização , Resveratrol/farmacologia , Resveratrol/química , Resveratrol/administração & dosagem , Resveratrol/farmacocinética , Nanofibras/química , Queimaduras/tratamento farmacológico , Cicatrização/efeitos dos fármacos , Animais , Colágeno/química , Povidona/química , Staphylococcus aureus/efeitos dos fármacos , Álcool de Polivinil/química , Humanos , Escherichia coli/efeitos dos fármacos , Ampicilina/farmacologia , Ampicilina/química , Ampicilina/farmacocinética , Ampicilina/administração & dosagem , Antibacterianos/farmacologia , Antibacterianos/química , Antibacterianos/administração & dosagem , Ratos , Biofilmes/efeitos dos fármacos , MasculinoRESUMO
Introduction: Previous publications have shown that STIM1, ORAI1, and KDM2B, are implicated in Ca2+ signaling and are highly expressed in various cancer subtypes including prostate cancer. They play multiple roles in cancer cell migration, invasion, and metastasis. In the current study we investigated the expression of the above biomarkers in circulating tumor cells from patients with metastatic prostate cancer. Methods: Thirty-two patients were enrolled in this study and CTCs' isolation was performed with Ficoll density gradient. Two different triple immunofluorescence stainings were conducted with the following combination of antibodies: CK/KDM2B/CD45 and CK/STIM1/ORAI1. Slides were analyzed using VyCAP microscopy technology. Results: CTC-positive patients were detected in 41% for (CK/KDM2B/CD45) staining and in 56% for (CK/STIM1/ORAI1) staining. The (CK+/KDM2B+/CD45-) and the (CK+/STIM1+/ORAI1+) were the most frequent phenotypes as they were detected in 85% and 94% of the CTC-positive patients, respectively. Furthermore, the expression of ORAI1 and STIM1 in patients' PBMCs was very low exhibiting them as interesting specific biomarkers for CTC detection. The (CK+/STIM1+/ORAI1+) phenotype was correlated to bone metastasis (p = 0.034), while the (CK+/STIM1+/ORAI1-) to disease relapse (p = 0.049). Discussion: STIM1, ORAI1, and KDM2B were overexpressed in CTCs from patients with metastatic prostate cancer. STIM1 and ORAI1 expression was related to disease recurrence and bone metastasis. Further investigation of these biomarkers in a larger cohort of patients will clarify their clinical significance for prostate cancer patients.
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KDM2B, a histone lysine demethylase, is expressed in a plethora of cancers. Earlier studies from our group, have showcased that overexpression of KDM2B in the human prostate cancer cell line DU-145 is associated with cell adhesion, actin reorganization, and improved cancer cell migration. In addition, we have previously examined changes of cytosolic Ca2+, regulated by the pore-forming proteins ORAI and the Ca2+ sensing stromal interaction molecules (STIM), via store-operated Ca2+ entry (SOCE) in wild-type DU-145. This study sought to evaluate the impact of KDM2B overexpression on the expression of key molecules (SGK1, Nhe1, Orai1, Stim1) and SOCE. Furthermore, this is the first study to evaluate KDM2B expression in circulating tumor cells (CTCs) from patients with prostate cancer. mRNA levels for SGK1, Nhe1, Orai1, and Stim1 were quantified by RT-PCR. Calcium signals were measured in KDM2B-overexpressing DU-145 cells, loaded with Fura-2. Blood samples from 22 prostate cancer cases were scrutinized for KDM2B expression using immunofluorescence staining and the VyCAP system. KDM2B overexpression in DU-145 cells increased Orai1, Stim1, and Nhe1 mRNA levels and significantly decreased Ca2+ release. KDM2B expression was examined in 22 prostate cancer patients. CTCs were identified in 45 % of these patients. 80 % of the cytokeratin (CK)-positive patients and 63 % of the total examined CTCs exhibited the (CK + KDM2B + CD45-) phenotype. To conclude, this study is the first to report increased expression of KDM2B in CTCs from patients with prostate cancer, bridging in vitro and preclinical assessments on the potentially crucial role of KDM2B on migration, invasiveness, and ultimately metastasis in prostate cancer.
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Objective: Cardiovascular diseases (CVD) are the leading cause of death globally. Metabolic syndrome (MtS) is a risk factor that increases the likelihood of CVD. The atherogenic index (AIP), calculated as the logarithm of the ratio of triglycerides (TG) to high-density lipoprotein cholesterol (HDL) cholesterol in plasma, is a valuable marker for highly atherogenic small dense low-density lipoprotein cholesterol particles. This study aimed to explore MtS prevalence and investigate the potential of using the AIP as a predictor for CVD risk factors in adults from the Qassim region of Saudi Arabia. Methods: The cross-sectional study enrolled 589 participants from public hospitals in nine major cities who completed a detailed questionnaire on health, diet, and lifestyle. Anthropometric measurements and some clinical parameters were measured. Results: The findings indicated a significant prevalence of MtS (37.5%) among participants from the Qassim Area, which was higher in males (39.9%) than females (34.9%). Nevertheless, a significant prevalence was shown for CVD risk factors among participants, with hyperglycemia (78.1%), hypertriglyceridemia (39.0%), hypo-HDL-cholesterolemia (38.9%), and hypertension (21.6%) being common. The AIP's performance in identifying CVD risk factors showed a receiver operating characteristic value of 0.909 (P < 0.001). The optimal cutoff value for the AIP was determined to be 0.468, demonstrating high sensitivity (84.8%) and specificity (78.6%). Conclusion: Incorporating AIP into clinical practice could enhance CVD risk prediction compared to using lipid profiles alone. These findings suggest that there is a high prevalence of MtS among adults in the Qassim region of Saudi Arabia. Further longitudinal studies are needed to recommend AIP as a robust tool for predicting CVD in clinical settings.
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Nanosized lanthanum oxide particles (La2O3) are commonly utilized in various industries. The potential health risks associated with La2O3 nanoparticles, cytotoxic effects at varying doses and time intervals, and the mechanisms behind their induction of behavioral changes remain uncertain and necessitate further investigation. Therefore, this study examined in vivo hepatotoxicity, considering the quantity (60, 150, and 300 mg/kg) and time-dependent induction of reactive oxygen species (ROS) over one week or 21 days. The mice received intraperitoneal injections of three different concentrations in Milli-Q water. Throughout the experiments, no physical changes or weight loss were observed among the groups. However, after 21 days, only the highest concentration showed signs of anxiety in the activity cage (p < 0.05). Subsequently, all animals treated with La2O3 NPs exhibited a significant loss of learning and memory recall using the Active Avoidances test, after 21 days (p < 0.001). Markers for anti-reactive oxygen species (ROS) such as superoxide dismutase (SOD) were significantly upregulated in response to all concentrations of NPs after seven days compared to the control group. This was confirmed by a significant increase in glutathione peroxidase (Gpx1) and pro-apoptotic Caspase-3 expression at the lowest and highest doses. Additionally, both transcription and protein levels of the anti-apoptotic BCL-2 surpassed P53 protein in a dosage-dependent manner, indicating activation of the primary anti-apoptosis pathway. After 21 days, P53 levels exceeded BCL-2 protein levels, confirming a significant loss of BCL-2 mRNA, particularly at the 300 mg/kg concentration. Furthermore, a higher transcription level of Caspase-3, SOD, and Gpx1 was observed, with the highest values detected at the 300 mg/kg concentration, indicating the activation of cell death. Histopathological analysis of the liver illustrated apoptotic bodies resulting from La2O3 NP concentration. The investigation revealed multiple inflammatory foci, cytoplasmic degeneration, steatosis, and DNA fragmentation consistent with increased damage over time due to higher concentrations. Blood samples were also analyzed to determine liver enzymatic changes, including alkaline phosphatase (ALP), alanine transaminase (ALT), aspartate aminotransferase (AST), and lipid profiles. The results showed significant differences among all La2O3 NP concentrations, with the most pronounced damage observed at the 300 mg/kg dose even after 21 days. Based on an animal model, this study suggests that La2O3 hepatotoxicity is likely caused by the size and shape of nanoparticles (NPs), following a dose and time-dependent mechanism that induces the production of reactive oxygen species and behavioral changes such as anxiety and memory loss.
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Doença Hepática Induzida por Substâncias e Drogas , Lantânio , Nanopartículas , Óxidos , Camundongos , Feminino , Animais , Espécies Reativas de Oxigênio/metabolismo , Caspase 3/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Nanopartículas/toxicidade , Apoptose , Fígado , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Superóxido Dismutase/metabolismo , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Estresse OxidativoRESUMO
Background: Antitumor research aims to efficiently target hepatocarcinoma cells (HCC) for drug delivery. Nanostructured lipid carriers (NLCs) are promising for active tumour targeting. Cell-penetrating peptides are feasible ligands for targeted cancer treatment. Methods: In this study, we optimized gefitinib-loaded NLCs (GF-NLC) for HCC treatment. The NLCs contained cholesterol, oleic acid, Pluronic F-68, and Phospholipon 90G. The NLC surface was functionalized to enhance targeting with the cRGDfK-pentapeptide, which binds to the αvß3 integrin receptor overexpressed on hepatocarcinoma cells. Results: GF-NLC formulation was thoroughly characterized for various parameters using differential scanning calorimetry and X-ray diffraction analysis. In-vitro and in-vivo studies on the HepG2 cell line showed cRGDfK@GF-NLC's superiority over GF-NLC and free gefitinib. cRGDfK@GF-NLC exhibited significantly higher cytotoxicity, growth inhibition, and cellular internalization. Biodistribution studies demonstrated enhanced tumour site accumulation without organ toxicity. The findings highlight cRGDfK@GF-NLC as a highly efficient carrier for targeted drug delivery, surpassing non-functionalized NLCs. These functionalized NLCs offer promising prospects for improving hepatocarcinoma therapy outcomes by specifically targeting HCC cells. Conclusion: Based on these findings, cRGDfK@GF-NLC holds immense potential as a highly efficient carrier for targeted drug delivery of anticancer agents, surpassing the capabilities of non-functionalized NLCs. This research opens up new avenues for effective treatment strategies in hepatocarcinoma.
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Carcinoma Hepatocelular , Neoplasias Hepáticas , Nanoestruturas , Humanos , Portadores de Fármacos/química , Carcinoma Hepatocelular/tratamento farmacológico , Gefitinibe , Distribuição Tecidual , Neoplasias Hepáticas/tratamento farmacológico , Nanoestruturas/química , Tamanho da Partícula , Lipídeos/químicaRESUMO
Psidium guajava L. is a small evergreen tree known for its magnificent medicinal and nutritional value. This study aimed to evaluate the nutritional profile and in vitro pharmacological potentialities of the different leaf extracts of four cultivars of Psidium guajava namely Surka chitti, Allahabad safeda, Karela, and Lucknow-49. The standard procedures of the Association of Official Analytical Chemists (AOAC) were followed to carry out the nutritional analysis and all of the cultivars recorded the presence of elements at a nominal range. The highest presence of phenols (125.77 mg GAE/g) and flavonoids (92.38 mg QE/g) in the methanolic leaf extract of the Karela cultivar was recorded. A wide range of minerals such as sodium, phosphorus, magnesium, zinc, and boron were recorded with a higher percentage in the Karela cultivar of Psidium guajava. In the enzyme inhibitory assays, Allahabad safeda showed potential inhibition with an IC50 of 113.31 ± 1.07, 98.2 ± 0.66 and 95.73 ± 0.39 µg/mL in α-amylase, α-glucosidase, and tyrosinase inhibition assays, respectively. The strong antioxidant effect was established by Lucknow-49 (IC50 of 74.43 ± 1.86 µg/mL) and Allahabad safeda (IC50 of 78.93 ± 0.46 µg/mL) for ABTS and DPPH assays, respectively. The ethyl acetate and methanolic leaf extracts of the Allahabad safeda cultivar showed better inhibition against Pseudomonas aeruginosa with an MIC of 14.84 and 28.69 µg/mL, respectively. A decent mean zone of inhibition was recorded in methanolic leaf extract that ranged from 21-25 mm in diameter against the tested bacterial strains (Proteus vulgaris, Bacillus subtilis, and P. aeruginosa). This is the first scientific report on the comparative and comprehensive analysis of indigenous guava cultivars to evidently shortlist the elite cultivars with enriched dietary nutrition and biological activities.
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Rice constitutes a foundational cereal and plays a vital role in the culinary sector. However, the detriments of abiotic stress on rice quality and productivity are noteworthy. Carotenoid cleavage oxygenases (CCO) hold vital importance as they enable the particular breakdown of carotenoids and significantly contribute towards the growth and response to abiotic stress in rice. Due to the insufficient information regarding rice CCOs and their potential role in abiotic stress, their utilization in stress-resistant genetic breeding remains limited. The current research identified 16 CCO genes within the Oryza sativa japonica group. These OsCCO genes can be bifurcated into three categories based on their conserved sequences: NCEDs (9-Cis-epoxycarotenoid dioxygenases), CCDs (Carotenoid cleavage dioxygenases) and CCD-like (Carotenoid cleavage dioxygenases-like). Conserved motifs were found in the OsCCO gene sequence via MEME analysis and multiple sequence alignment. Stress-related cis-elements were detected in the promoter regions of OsCCOs genes, indicating their involvement in stress response. Additionally, the promoters of these genes had various components related to plant light, development, and hormone responsiveness, suggesting they may be responsive to plant hormones and involved in developmental processes. MicroRNAs play a pivotal role in the regulation of these 16 genes, underscoring their significance in rice gene regulation. Transcriptome data analysis suggests a tissue-specific expression pattern for rice CCOs. Only OsNCED6 and OsNCED10 significantly up-regulated during salt stress, as per RNA seq analyses. CCD7 and CCD8 levels were also higher in the CCD group during the inflorescence growth stage. This provides insight into the function of rice CCOs in abiotic stress response and identifies possible genes that could be beneficial for stress-resistant breeding.
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Nanoparticles are widely used in the pharmaceutical, agriculture, and food processing industries. In this study, we have synthesized green lead nanoparticles (gPbNPs) by using an extract of Ziziphus spina-christi leaves and determined their cytotoxic and apoptotic effect on the human breast cancer MDA-MB-231 cell line. gPbNPs were characterized by using X-ray diffraction (XRD), energy dispersive X-ray (EDX) scanning electron microscope (SEM), and transmission electron microscope (TEM). The toxicity of gPbNPs was determined on the MDA-MB-231 cell line using MTT and NRU assays and as a result cell viability was reduced in a concentration-dependent manner. MDA-MB-231 cells were more sensitive at the highest concentration of gPbNPs exposure. In this experiment, we observed the production of intracellular ROS in cells, and induction of caspase 3/7 was higher in cells at 42 µg/ml of gPbNPs. Moreover, the Bax gene was upregulated and the Bcl-2 gene was downregulated and increased caspase 3/7 activity confirmed the apoptotic effect of gPbNPs in cells. Our observation showed that gPbNPs induced cell toxicity, increased generation of intracellular ROS, and gene expression of Bcl-2 and Bax in the MDA-MB-231 cell line. In conclusion, these findings demonstrated that gPbNPs executed toxic effects on the MDA-MB-231 cell line through activating caspase 3/7 activity.
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Electrospun composite nanofiber scaffolds are well known for their bone and tissue regeneration applications. This research is focused on the development of PVP and PVA nanofiber composite scaffolds enriched with hydroxyapatite (HA) nanoparticles and alendronate (ALN) using the electrospinning technique. The developed nanofiber scaffolds were investigated for their physicochemical as well as bone regeneration potential. The results obtained from particle size, zeta potential, SEM and EDX analysis of HA nanoparticles confirmed their successful fabrication. Further, SEM analysis verified nanofiber's diameters within 200-250 nm, while EDX analysis confirmed the successful incorporation of HA and ALN into the scaffolds. XRD and TGA analysis revealed the amorphous and thermally stable nature of the nanofiber composite scaffolds. Contact angle, FTIR analysis, Swelling and biodegradability studies revealed the hydrophilicity, chemical compatibility, suitable water uptake capacity and increased in-vitro degradation making it appropriate for tissue regeneration. The addition of HA into nanofiber scaffolds enhanced the physiochemical properties. Additionally, hemolysis cell viability, cell adhesion and proliferation by SEM as well as confocal microscopy and live/dead assay results demonstrated the non-toxic and biocompatibility behavior of nanofiber scaffolds. Alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP) assays demonstrated osteoblast promotion and osteoclast inhibition, respectively. These findings suggest that developed HA and ALN-loaded PVP/PVA-ALN-HA nanofiber composite scaffolds hold significant promise for bone regeneration applications.
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Electrospinning is a versatile method for fabrication of précised nanofibrous materials for various biomedical application including tissue engineering and drug delivery. This research is aimed to fabricate the PVP/PVA nanofiber scaffold by novel electrospinning technique and to investigate the impact of process parameters (flow rate, voltage and distance) and polymer concentration/solvent combinations influence on properties of electrospun nanofibers. The in-vitro and in-vivo degradation studies were performed to evaluate the potential of electrospun PVP/PVA as a tissue engineering scaffold. The solvents used for electrospinning of PVP/PVA nanofibers were ethanol and 90% acetic acid, optimized with central composite design via Design Expert software. NF-2 and NF-35 were selected as optimised nanofiber formulation in acetic acid and ethanol, and their characterization showed diameter of 150-400 nm, tensile strength of 18.3 and 13.1 MPa, respectively. XRD data revealed the amorphous nature, and exhibited hydrophilicity (contact angles: 67.89° and 58.31° for NF-2 and NF-35). Swelling and in-vitro degradability studies displayed extended water retention as well as delayed degradation. FTIR analysis confirmed solvent-independent interactions. Additionally, hemolysis and in-vitro cytotoxicity studies revealed the non-toxic nature of fabricated scaffolds on RBCs and L929 fibroblast cells. Subcutaneous rat implantation assessed tissue response, month-long biodegradation, and biocompatibility through histological analysis of surrounding tissue. Due to its excellent biocompatibility, this porous PVP/PVA nanofiber has great potential for biomedical applications.
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The SARS-CoV-2 virus gains entry into human cells by exploiting the angiotensin-converting enzyme 2 (ACE2), a key component known as the spike protein (S), as a point of entry. Initially, SARS-CoV-2 suppresses the natural function of ACE2, leading to a gradual decline in cell health. Additionally, individuals with cancer are considered more susceptible to COVID-19. This study investigates the expression patterns of ACE2 in colorectal cancer (CRC) patients with and without a history of COVID-19 infection. RT-PCR was used to analyze samples from both cancerous and adjacent non-affected colorectal tissues of 47 CRC patients, comprising two groups: 24 CRC patients with no history of COVID-19 and 23 CRC patients with a recent history of COVID-19 infection. Epithelial CR cells were isolated from both types of tissues and cultured to evaluate cell adhesion. Immunohistochemistry analyses were conducted to examine ACE2 protein expression using various ACE2 antibodies for both cell types. The study revealed ACE2 mRNA expression in all CRC tissues of patients with and without a history of COVID-19. ACE2 expression was significantly higher in CRC patients without a history of COVID-19. Notably, the non-affected colorectal cancer (NACRC) tissues of patients without a history of COVID-19 also showed ACE2 expression, whereas no ACE2 expression was detected in the biopsies of CRC patients with a positive COVID-19 history. ACE2 antibodies were employed to validate ACE2 protein expression at the mRNA level. COVID-19 appears to downregulate ACE2 expression in both CRC and NACRC tissues of CRC patients with a positive history of COVID-19 infection.
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COVID-19 , Neoplasias Colorretais , Humanos , SARS-CoV-2/genética , Enzima de Conversão de Angiotensina 2/genética , RNA Mensageiro/genética , Peptidil Dipeptidase A/genética , Peptidil Dipeptidase A/metabolismoRESUMO
The anti-inflammatory effect of the ethyl acetate extract of F. microcarpa bark (EAFMB) was investigated in acute and chronic (21 days) inflammation induced in Wistar albino rats. EAFMB (200 mg/kg b.w.) exhibited comparable anti-inflammatory effects to the reference drug, with a reduction of 59.48% at 4 h in acute inflammation and 83.96% on day 21 in chronic inflammation. Bioassay-guided fractionation using DPPH radical scavenging activity led to isolating and identifying three compounds from EAFMB: oleanolic acid, catechin, and p-hydroxycinnamic acid. All these compounds demonstrated the concentration-dependent inhibition of COX enzymes and the protection of egg albumin from heat-induced denaturation. Catechin exhibited the highest COX inhibition (COX-1 and COX-2 IC50 = 9.02 and 50.38 µM, respectively) and anti-denaturation effect (IC50 = 27.13 µg/mL) compared to oleanolic acid and p-hydroxycinnamic acid. These isolated compounds are likely responsible for the anti-inflammatory activities of F. microcarpa bark.
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Background: The purpose of this study is to analyzed the involvement of chorein in microtubules organization of three types of malignant; rhabdomyosarcoma tumor cells (ZF), rhabdomyosarcoma cells (RH30), and rhabdomyosarcoma cells (RD). ZF are expressing high chorein levels. Previous studies revealed that chorein protein silencing in ZF tumor cells persuaded apoptotic response followed by cell death. In addition, in numerous malignant and non-malignant cells this protein regulates actin cytoskeleton structure and cellular signaling. However, the function of chorein protein in microtubular organization is yet to be established. Methods: In a current research study, we analyzed the involvement of chorein in microtubules organization by using three types of malignant rhabdomyosarcoma cells. We have applied confocal laser-scanning microscopy to analyze microtubules structure and RT-PCR to examine cytoskeletal gene transcription. Results: We report here that in rhabdomyosarcoma cells (RH30), chorein silencing induced disarrangement of microtubular network. This was documented by laser scanning microscopy and further quantified by FACS analysis. Interestingly and in agreement with previous reports, tubulin gene transcription in RH cells was unchanged upon silencing of chorein protein. Equally, confocal analysis showed minor disordered microtubules organization with evidently weakened staining in rhabdomyosarcoma cells (RD and ZF) after silencing of chorein protein. Conclusion: These results disclose that chorein silencing induces considerable structural disorganization of tubulin network in RH30 human rhabdomyosarcoma tumor cells. Additional studies are now needed to establish the role of chorein in regulating cytoskeleton architecture in tumor cells.
Assuntos
Rabdomiossarcoma , Tubulina (Proteína) , Proteínas de Transporte Vesicular , Humanos , Citoesqueleto de Actina , Citoesqueleto/genética , Microtúbulos , Rabdomiossarcoma/genética , Linhagem Celular Tumoral , Proteínas de Transporte Vesicular/genéticaRESUMO
Green synthesis of nanoparticles has drawn huge attention in the last decade due to their eco-friendly, biocompatible nature. Phyto-assisted synthesis of metallic nanoparticles is widespread in the field of nanomedicine, especially for antimicrobial and anticancer activity. Here in the present research work, investigators have used the stem extract of the Himalayan plant Hippophae rhamnoides L, for the synthesis of copper nanoparticles (CuNPs). The synthesized of CuNPs were analyzed by using sophisticated instruments, i.e., Fourier transform infrared spectroscopy (FTIR), UV-Vis spectroscopy, X-ray diffraction (XRD), high-performance liquid chromatography (HPLC), and scanning electron microscope (SEM). The size of the synthesized CuNPs was varying from 38 nm to 94 nm which were mainly spherical in shape. Further, the potential of the synthesized CuNPs was evaluated as an anticancer agent on the Hela cell lines, by performing an MTT assay. In the MTT assay, a concentration-dependent activity of CuNPs demonstrated the lower cell viability at 100 µg/mL and IC50 value at 48 µg/mL of HeLa cancer cell lines. In addition to this, apoptosis activity was evaluated by reactive oxygen species (ROS), DAPI (4',6-diamidino-2-phenylindole) staining, Annexin V, and Propidium iodide (PI) staining, wherein the maximum ROS production was at a dose of 100 µg per mL of CuNPs with a higher intensity of green fluorescence. In both DAPI and PI staining, maximum nuclear condensation was observed with 100 µg/mL of CuNPs against HeLa cell lines.
RESUMO
Nanoparticles have shown promising potential for efficient drug delivery, circumventing biological interferences like immunological and renal clearance and mechanical and enzymatic destruction. However, a handful of research papers have questioned the biomedical use of metal-based nanoparticles like cadmium telluride quantum dots (CdTe-QDs) for their cytotoxic, genotoxic, and carcinogenic potential. Herein, we examined the effects of CdTe-QD NPs on gene expression profile of hepatocellular carcinoma (Huh-7) cell line. Huh-7 cells were treated with CdTe-QD NPs (10 µg/ml for 6, 12, and 24 hours, and 25 µg/ml for 6 and 12 hours), and transcriptomic analysis was performed using microarray to evaluate the global gene expression profile. Differential expressed genes (DEGs) were observed for both the doses (10 and 25 µg/ml) of CdTe-QD NPs at different time points. Gene ontology (GO) analysis revealed that genes involved in molecular function of cell cycle, organizational injury and abnormalities, cell death and survival, gene expression, cancer, organismal survival, and cellular development were differentially expressed. Overall, we have demonstrated differential expression of several genes, involved in maintaining cell survival, metabolism, and genome integrity. These findings were confirmed by RT-qPCR study for some canonical pathway genes signifying possible implication in NP toxicity-mediated cell survival and inhibition of cell death.