Determination Of Viability Research Articles

Determinants of Financial Viability of Forest Concession in Brazilian Amazon

This study aimed to assess the financial viability of forest concessions in the state of Pará, Brazil. Two Forest Management Units (FMUs) were analyzed: FMU-2, located in Saracá-Taquera National Forest, and FMU-3, located in Caxiuanã National Forest. Financial indicators were evaluated under different timber productivity scenarios (20 m3/ha and 25.8 m3/ha). At a logging intensity of 20 m3/ha, FMU-2 was not financially viable. However, both FMUs were financially viable at 25.8 m3/ha. Sensitivity analysis demonstrated that FMU-2 requires a logging intensity of over 22 m3/ha and a sawmill yield efficiency of at least 45%. The ideal cost for sawn wood would be USD 226.53/m3, with royalties of USD 16.00/m3. FMU-3 consistently demonstrated positive financial results, despite fluctuations in production costs. The financial viability of investing in forest concessions in state of Pará depends largely on forest productivity, sawmill yield, royalties, and sawn wood costs.

Antimicrobial and Cytotoxic Potential of Eucalyptus Essential Oil-Based Nanoemulsions for Mouthwashes Application.

Objective: An eucalyptus essential oil-based nanoemulsion was produced and evaluated for its antimicrobial properties against Streptococcus mutans and its cytotoxicity in the surface mucous cells of rabbits. Methods: The essential oil-based nanoemulsion was synthesized with two species of eucalyptus-Eucalyptus citriodora and Eucalyptus globulus-followed by physicochemical characterization and the determination of antimicrobial activity and cell viability. Subsequently, the mouthwash formulations (fluoride and fluoride-free) were functionalized with the nanoemulsion, and their in vitro antimicrobial actions were evaluated against S. mutans. Results: The nanoemulsion presented an average particle size of around 100 nm, a polydispersity index close to 0.3, a zeta potential between -19 and -30 mV, a pH close to 7, a spherical shape, and a cell viability above 50%. The antimicrobial activity analysis showed that the nanoemulsion was effective in the control of S. mutans. The mouthwashes functionalized with the nanoemulsion also presented bacteriostatic and bactericidal properties. Conclusions: The bio-based material produced with eucalyptus essential oil presented adequate physicochemical characteristics, with the potential to be used as an innovative material in preventive dentistry, contributing to the maintenance of oral and systemic health.

Assessment of the In Vitro Phosphatidylinositol Glycan Class A (PIG-A) Gene Mutation Assay Using Human TK6 and Mouse Hepa1c1c7 Cell Lines.

Gene mutations linked to diseases like cancer may be caused by exposure to environmental chemicals. The X-linked phosphatidylinositol glycan class A (PIG-A) gene, required for glycosylphosphatidylinositol (GPI) anchor biosynthesis, is a key target locus for in vitro genetic toxicity assays. Various organisms and cell lines may respond differently to genotoxic agents. Here, we compared the mutagenic potential of directly genotoxic ethyl methane sulfonate (EMS) to metabolically activated pro-mutagenic polycyclic aromatic hydrocarbons (PAHs). The two classes of mutagens were compared in an in vitro PIG-A gene mutation test using the metabolically active murine hepatoma Hepa1c1c7 cell line and the human TK6 cell line, which has limited metabolic capability. Determination of cell viability is required for quantifying mutagenicity. Two common cell viability tests, the MTT assay and propidium iodide (PI) staining measured by flow cytometry, were evaluated. The MTT assay overestimated cell viability in adherent cells at high benzo[a]pyrene (B[a]P) exposure concentrations, so PI-based cytotoxicity was used in calculations. The spontaneous mutation rates for TK6 and Hepa1c1c7 cells were 1.87 and 1.57 per million cells per cell cycle, respectively. TK6 cells exposed to 600 µM and 800 µM EMS showed significantly higher mutation frequencies (36 and 47 per million cells per cell cycle, respectively). Exposure to the pro-mutagen benzo[a]pyrene (B[a]P, 10 µM) did not increase mutation frequency in TK6 cells. In Hepa1c1c7 cells, mutation frequencies varied across exposure groups (50, 50, 29, and 81 per million cells per cell cycle when exposed to 10 µM B[a]P, 5-methylcholanthrene (5-MC), chrysene, or 16,000 µM EMS, respectively). We demonstrate that the choice of cytotoxicity assay and cell line can determine the outcome of the Pig-A mutagenesis assay when assessing a specific mutagen.

Combining Killed Vaccine Candidate with Different Adjuvants to Determine Prophylactic Potential against Leishmaniasis.

Visceral Leishmaniasis is a serious public health problem caused by Leishmania species parasites. Approximately 500 thousand people get Visceral Leishmaniasis (VL) every year. An effective and reliable vaccine against the disease has still not been formulated. Choosing the right adjuvant is important to increase immunogenicity in vaccines prepared with total antigens. In this study, we investigate the ideal adjuvant for use in vaccine formulations against VL. For this purpose, Leishmania antigens (FTLA) obtained from L. infantum parasites by the freeze-thaw method and three different adjuvants (alum-saponin and calcium phosphate) were used. The effectiveness of the formulations was investigated in vitro by cell viability analysis and determination of nitric oxide and cytokine production abilities in J774 macrophage cells. According to the study results, it was determined that formulations prepared with calcium phosphate produced 72% more NO and approximately 7.2 times more IL-12 cytokine. The results obtained showed that calcium phosphate salts can be used as ideal adjuvants in vaccine research against leishmaniasis.

Sources of Variability in Determinations of Brewing Yeast Total Cell Counts and Viability Obtained by Microscopy

Brewers utilizing yeast cell counts and determinations of yeast viability obtained by microscopy require precise results for calculating the yeast pitch to produce fermentations with the highest quality and consistency. The steps in obtaining results with the desired precision include identifying, quantifying and controlling the sources of variation. In this study, we used simulated cell counting, manual cell counting, and semi-automated cell counting to explore sources of variation contributing to the variability of yeast cell counts and yeast viability determinations. The results showed that the variability associated with counting different fields of view contributed most to the variability in yeast cell counts and determinations of yeast viability. Understanding the sources and amounts of variation in cell counts will help brewers direct their efforts toward obtaining the highest quality determinations of yeast cell counts and viability.

Staining to machine learning: An emerging technology for determination of microalgal cell viability

Staining to machine learning: An emerging technology for determination of microalgal cell viability

Poster 336: Reliability of Preoperative MRI in the Prediction of ACL Tear Type

Objectives: Anterior cruciate ligament (ACL) tears are among the most frequent ligament injuries in the United States. Most of these injuries are treated surgically, depending on patient-specific factors. Management of these injuries includes nonoperative management with stepwise rehabilitation or surgical treatment with ACL repair or reconstruction. Patient goals, a clinical evaluation, and preoperative MRI imaging are generally assessed to inform the choice of treatment. Classification of ACL tear-type based on stump morphology is critical to preoperative planning, as it can determine a patient’s eligibility for ACL preservation techniques. We aim to determine the efficacy of preoperative magnetic resonance imaging (MRI) in predicting ruptured ACL stump morphology and thus assist in preoperative planning and decision-making. Methods: This retrospective observational study utilized chart review from the case log of a single surgeon at an urban tertiary institution. The review identified patients who underwent the Bridge Enhanced ACL Repair (BEAR) procedure between January 1, 2022 and August 31, 2023. The inclusion criteria included patients who underwent a primary ACL repair with the BEAR technique and had adequate intraoperative and MRI images available for review. Patients excluded were those with inadequate intraoperative imaging or MRI imaging that was unavailable for review. For each patient included, the two senior authors assessed the morphology of the ACL stump on MRI (sagittal T2, coronal T2) and intraoperative arthroscopy images in a blinded manner and assigned a classification grade to each. The MRI-based morphology and the intraoperative image morphology of the tears were classified based on a system validated by DeFelice et al, which includes types 1 to 5 based on the location of the tear from most proximal to most distal. The correlation between the preoperative MRI assessment and the intraoperative arthroscopic assessment was interrogated using simple linear regression quantified with the Pearson correlation coefficient. Results: A total of 35 patients that underwent the BEAR procedure were identified. One patient was excluded due to inadequate intraoperative images. Of the remaining patients, 13 (38.2%) were male and 21 (61.8%) were female. The average patient age was 31, with a median age of 28. Most patients were injured while skiing (32.4%), while the second and third most prevalent mechanisms of injury were nonsports-related (23.4%) and soccer injuries (17.6%), respectively. Of the MRI tears that were analyzed, 9 (26.5%) were classified as type 1, 19 (55.9%) were type 2, and 5 (14.7%) were type 3. There were no tears identified as types 4 or 5 tears. Of the arthroscopic images that were analyzed, 21 (61.8%) were classified as type 1, 10 (29.4%) were type 2, 3 (8.8%) were type 3. Similarly, there were no tears identified as types 4 or 5. MRI accurately predicted the ACL tear type seen arthroscopically 55.9% (19/34) of the time. Of the (15) cases in which MRI did not correlate with the arthroscopic tear type, the MRI classification grade was within 1 grade of the arthroscopic grade 93.3% of the time (14/15). In the cases in which MRI was not correct in its grading, 93.3% of them (14/15) were classified as too high of a grade (MRI perceiving the tear to be more distal than it was in actuality). The Pearson correlation coefficient assessing the correlation between MRI identified tear type and arthroscopy identified tear type was 0.56. Conclusions: MRI accurately predicted the ACL tear type seen arthroscopically in only 55.9% of patients. In >90% of incorrect predictions, arthroscopy determined the ACL tear to be one classification more proximal than predicted by MRI. Our results demonstrate that while preoperative MRI can be useful in identifying approximate ACL tear location, it is essential to examine the ACL at time of arthroscopy for final determination of ACL stump size and viability for ACL restoration procedures. Moreover, a greater percentage of patients may be eligible for an ACL restoration procedure than predicted by preoperative MRI, highlighting the importance of discussing both ACL repair and ACL reconstruction with patients preoperatively. [Figure: see text]

A novel role for the peptidyl-prolyl cis-trans isomerase Cyclophilin A in DNA-repair following replication fork stalling via the MRE11-RAD50-NBS1 complex

Cyclosporin A (CsA) induces DNA double-strand breaks in LIG4 syndrome fibroblasts, specifically upon transit through S-phase. The basis underlying this has not been described. CsA-induced genomic instability may reflect a direct role of Cyclophilin A (CYPA) in DNA repair. CYPA is a peptidyl-prolyl cis-trans isomerase (PPI). CsA inhibits the PPI activity of CYPA. Using an integrated approach involving CRISPR/Cas9-engineering, siRNA, BioID, co-immunoprecipitation, pathway-specific DNA repair investigations as well as protein expression interaction analysis, we describe novel impacts of CYPA loss and inhibition on DNA repair. We characterise a direct CYPA interaction with the NBS1 component of the MRE11-RAD50-NBS1 complex, providing evidence that CYPA influences DNA repair at the level of DNA end resection. We define a set of genetic vulnerabilities associated with CYPA loss and inhibition, identifying DNA replication fork protection as an important determinant of viability. We explore examples of how CYPA inhibition may be exploited to selectively kill cancers sharing characteristic genomic instability profiles, including MYCN-driven Neuroblastoma, Multiple Myeloma and Chronic Myelogenous Leukaemia. These findings propose a repurposing strategy for Cyclophilin inhibitors.

Determination of Cell Viability, Apoptotic and Necrotic Activities of Boswellia serrata Extract in Liver Cancer (HepG2) and Breast Cancer (MCF7) Cell Lines

Cancer is a very important disease that has existed for a very long time in the history of humanity and poses a great threat to humanity today. Today, cancer remains a global health problem. The scientific world has introduced many innovative treatment approaches, but due to the complexity of cancer cells and their resistance to treatment, significant progress has been made in the fight against cancer, but a definitive cure has not yet been found. Boswellia serrata has been used in many parts of the world for a variety of ailments from ancient times to the present day, comes from the Burseraceae family and is commonly known as white gum. B. serrata is a resin tree with very small and few leaves, as it is a plant species that grows as a small tree or shrub 4-6 meters high in the desert. In this study, B.serrata resin was obtained commercially in Gaziantep province and obtained by extraction in methanol and hexane solvents. The cytotoxic activity of B. serrata resin extract was determined by MTT Assay (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide). Apoptotic/necrotic activity was determined by double staining (Hoechst 33342 and propidium iodide). Human breast cancer (MCF7) and human liver cancer (HepG2) cell lines were used in these experiments.

The Potential Of Superoxide Dismutase Enzyme From Tomato Fruit (Solanum Lycopersicum) To Repair Collagen Damage In 3t3 Fibroblast Cells Exposed To Ultraviolet A Radiation

UV A radiation can cause photoaging to the skin which is show by a decrease in the amount of collagen and death of fibroblast cell. This study aimed to determine the SOD enzyme activity of tomatoes (Solanum lycopersycum Mill.) influences the increase of viability of fibroblast cell and deposition of collagen. The study design used a completely randomized design (CRD) with 11 treatments and 3 repetitions. The procedures of this study include: (1) extracting SOD enzymes (2) making bradford reagents (3) making standard protein solutions (4) measuring total protein content (5) testing the antioxidant activity of SOD enzymes (6) preparing 3T3 fibroblast cells (7) Cell exposure 3T3 fibroblasts with UV A rays (8) preparation of tomato SOD enzyme stock solutions (9) determination of cell viability and collagen deposition. Data analized by normality test, homogeneity test, one way ANOVA test, tuckey test and dunnet test. The results of SOD enzyme activity showed the highest percent inhibition was 77.83%. Highest value of percent viability of fibroblast cells that were exposed to UV A rays after given 90% SOD enzyme treatment was 233.00% and collagen deposition was 157.67%. The results showed that variations in the SOD enzyme concentration of tomatoes had a significant effect on cell viability and collagen deposition in fibroblast cells. with a value of sig = 0,000 (p <α). Based on the result, we can concluded that SOD enzyme from tomatoe fruit show significant activity as anti-photoaging.

Effect of fructose on anthracycline and HER-2 blocking agent mediated cardiotoxicity through NLRP3 and MyD88 mediated pathways: New roles of sweeteners in cardio-oncology.

e24020 Background: Combinatorial therapy based on anthracyclines and HER-2 blocking agents increases the risk of cardiomyopathy in women with breast cancer. Fructose is a monosaccharide associated with high risk of diabetes, non-alcoholic fatty liver disease, atherosclerosis and visceral obesity. A high fructose diet is a potential cardiometabolic risk factor and may increase cardiac risk in women with breast cancer treated with cardiotoxic drugs. In this study, we highlighted the role of sweeteners fructose in the worsening of cardiotoxicity induced by combinatorial therapy anthracycline-HER2-blocking agents. Methods: Human cardiomyocytes were pre-exposed with doxorubicin combined to trastuzumab (at 300 and 1000 nM) for 48 and 72h alone or combined to fructose at 0.25, 1 and 2.5 mM. After the incubation period, we performed the following tests: determination of cell viability, through mitochondrial dehydrogenase activity, study of lipid peroxidation (quantifying cellular Malondialdehyde and 4-hydroxynonenal), intracellular Ca2+ homeostasis. Moreover, pro-inflammatory studied were also performed (activation of NLRP3 inflammasome; expression of peroxisome proliferator-activated receptor-α; transcriptional activation of p65/NF-κB and secretion of cytokines involved in cardiotoxicity (Interleukins 1β, 8, 6, CXCL-12). Results: High fructose increases significantly cell mortality of cardiomyocytes exposed to doxorubicin and trastuzumab through the activation of NLRP-3 and Myd-88 expression. Intracellular Ca++ levels were also increased of 4 times compared to non-fructose exposed cardiomyocytes; MDA and 4-HNA levels were increased of 48,3 and 51,2 % compared to non-fructose exposed cells (p<0.001). Fructose increases IL-1, IL-6 and CXCL-12 expression in cardiomyocytes in a concentration dependent manner, compared to doxorubicin-trastuzumab group (p<0.05). Conclusions: Data of the present study indicate that fructose induces a pro-inflammatory phenotype in human cardiac cells exposed to anticancer cardiotoxic drugs, providing even more data on the harmful role of fructose as a key player of cardiotoxic phenomena.

FRUCTOSE EXACERBATES ANTHRACYCLINE AND HER–2 BLOCKING AGENT MEDIATED CARDIOTOXICITY THROUGH NLRP3 AND MYD88 MEDIATED PATHWAYS: NEW ROLES OF SWEETENERS IN CARDIO–ONCOLOGY

Abstract Background Combinatorial therapy based on anthracyclines and HER–2 blocking agents increases the risk of cardiomyopathy in women with breast cancer. Fructose is a monosaccharide associated with high risk of diabetes, non–alcoholic fatty liver disease, atherosclerosis and visceral obesity. A high fructose diet is a potential cardiometabolic risk factor and may increase cardiac risk in women with breast cancer treated with cardiotoxic drugs. Purpose In this study, we highlighted the role of sweeteners fructose in the worsening of cardiotoxicity induced by combinatorial therapy anthracycline–HER2–blocking agents. Methods Human cardiomyocytes were pre–exposed with doxorubicin combined to trastuzumab (at 300 and 1000 nM) for 48 and 72h alone or combined to fructose at 0.25, 1 and 2.5 mM. After the incubation period, we performed the following tests: determination of cell viability, through mitochondrial dehydrogenase activity, study of lipid peroxidation (quantifying cellular Malondialdehyde and 4–hydroxynonenal), intracellular Ca2+ homeostasis. Moreover, pro–inflammatory studied were also performed (activation of NLRP3 inflammasome; expression of peroxisome proliferator–activated receptor–α; transcriptional activation of p65/NF–κB and secretion of cytokines involved in cardiotoxicity (Interleukins 1β, 8, 6, CXCL–12). Results High fructose increases significantly cell mortality of cardiomyocytes exposed to doxorubicin and trastuzumab thorugh the activation of NLRP–3 and Myd–88 expression. Intracellular Ca++ levels were also increased of 4 times compared to non–fructose exposed cardiomyocytes; MDA and 4–HNA levels were increased of 48,3 and 51,2 % compared to non–fructose exposed cells (p<0.001). Fructose increases IL–1, IL–6 and CXCL–12 expression in cardiomyocytes in a concentration dependent manner, compard to doxorubicin–trastuzumab group (p<0.05). Conclusion Data of the present study indicate that fructose induces a pro–inflammatory phenotype in human cardiac cells exposed to anticancer cardiotoxic drugs, providing even more data on the harmful role of fructose as a key player of cardiotoxic phenomena.

PRG4 represses the genesis and metastasis of osteosarcoma by inhibiting PDL1 expression

BackgroundOsteosarcoma is originated from skeletal system. Recombinant human proteoglycan 4 (rhPRG4) can inhibit cell proliferation and migration in multiple cancers. This research is designed to dig out the role and mechanism of PRG4 in osteosarcoma. MethodsHuman osteosarcoma cell lines, MG63 and 143B, were transfected with programmed death 1 (PD-L1) overexpression vectors and/or treated with 20, 50, and 100 μg/mL rhPRG4, followed by the determination of cell viability, colony formation, sphere formation, invasion, migration, apoptosis, and the expressions of matrix metalloproteinases (MMPs), PD-L1 and apoptosis-related proteins. Tumor-bearing mouse models were constructed by injection of 143B cells and treatment of anti-PD-L1 antibody and/or adenovirus PRG4 (AdPRG4). Tumor volume was monitored, and immunohistochemical location of Ki67 was performed. Expressions of MMPs, transforming growth factor-β (TGF-β), PD-L1, and epithelial mesenchymal transition (EMT)-related proteins were measured in tumors. ResultsRhPRG4 (20, 50, and 100 μg/mL) inhibited the viability, colony formation, sphere formation, invasion, migration, and the expressions of MMP2, MMP9 and Bcl2 in osteosarcoma cells, while promoting cell apoptosis as well as Bax and c-caspase3 expressions, at a dose-dependent manner; by contrast, PD-L1 overexpression reversed the above effects of 100 μg/mL rhPRG4. AdPRG4 or anti-PD-L1 antibody decreased tumor volume, number of pulmonary metastasis nodule, Ki67 location, and expressions of TGF-β, PD-L1, MMP2, MMP9, Vimentin, and Snail, but increased E-cadherin expression in tumor cells. Moreover, anti-PD-L1 antibody and AdPRG4 together functioned more effectively than them alone in reducing tumor burden. ConclusionPRG4 represses the genesis and metastasis of osteosarcoma via inhibiting PD-L1 expression, and AdPRG4 enhances the effectiveness of anti-PD-L1 therapy.

How precisely can x-ray predict the viability of wild flower plant seeds?

The detection of seed viability is an important step in seed production, as well as in conservation and restoration practice. Due to random natural events, the quality and viability of seeds of wild flowering species vary substantially, and hence a quick and reliable method for seed viability assessment is desirable. X-rays provide information about the internal structures of a seed and therefore show promise for detection of viability and even germination capacity. Seeds of 207 accessions of 176 wild flowering plant species were x-rayed and the viability results were compared with combined germination-TZ test results. Of special interest was whether there are certain plant families for which x-ray is an appropriate method for viability detection, considering correlations with seed internal morphology, seed mass and/or shape. The comparison revealed a strong correspondence between viability determination by combined germination-TZ tests and by x-ray analysis. According to taxonomy and seed type, two main groups could be distinguished, that differed significantly in viability detection by x-ray and combined germination-TZ test. Whereas the evaluation of little/non-endospermic seeds gave approximately identical results, there was greater discrepancy for of endospermic seeds. Seeds of different sizes and shapes were evaluated similarly with both methods. Especially for little/non-endospermic seeds, x-ray can provide a useful and quick tool for viability detection, whereas for endospermic seeds, further research is needed. For the commercial seed industry, viability detection via x-ray should be the first step before further vigour testing is performed.

Functional compensation in a savanna scavenger community.

Functional redundancy, the potential for the functional role of one species to be fulfilled by another, is a key determinant of ecosystem viability. Scavenging transfers huge amount of energy through ecosystems and is, therefore, crucial for ecosystem viability and healthy ecosystem functioning. Despite this, relatively few studies have examined functional redundancy in scavenger communities. Moreover, the results of these studies are mixed and confined to a very limited range of habitat types and taxonomic groups. This study attempts to address this knowledge gap by conducting a field experiment in an undisturbed natural environment assessing functional roles and redundancy in vertebrate and invertebrate scavenging communities in a South African savanna. We used a large-scale field experiment to suppress ants in four 1 ha plots in a South African savanna and paired each with a control plot. We distributed three types of small food bait: carbohydrate, protein and seed, across the plots and excluded vertebrates from half the baits using cages. Using this combination of ant suppression and vertebrate exclusion, allowed us explore the contribution of non-ant invertebrates, ants and vertebrates in scavenging and also to determine whether either ants or vertebrates were able to compensate for the loss of one another. In this study, we found the invertebrate community carried out a larger proportion of overall scavenging services than vertebrates. Moreover, although scavenging was reduced when either invertebrates or vertebrates were absent, the presence of invertebrates better mitigated the functional loss of vertebrates than did the presence of vertebrates against the functional loss of invertebrates. There is a commonly held assumption that the functional role of vertebrate scavengers exceeds that of invertebrate scavengers; our results suggest that this is not true for small scavenging resources. Our study highlights the importance of invertebrates for securing healthy ecosystem functioning both now and into the future. We also build upon many previous studies which show that ants can have particularly large effects on ecosystem functioning. Importantly, our study suggests that scavenging in some ecosystems may be partly resilient to changes in the scavenging community, due to the potential for functional compensation by vertebrates and ants.

Epithelial-Mesenchymal Transformation Promotes the Progression of Hepatocellular Carcinoma through NF-κB/MMP9 Axis.

Primary liver cancer (PHC) stands as one of the most prevalent malignant diseases in clinical settings. Studies have indicated that transcatheter arterial chemoembolization (TACE) treatment exhibits superior clinical outcomes, potentially increasing the complete necrosis rate in patients with PHC. A correlation exists between the clinical outcomes of TACE surgery and the process of epithelial-mesenchymal transition (EMT), yet the underlying mechanism remains a mystery. Hence, it is crucial to investigate the impact and mechanism of EMT on hepatocellular carcinoma (HCC). Retrospectively, patients with advanced liver cancer who underwent TACE were selected and categorized into two groups based on the assessment of clinical efficacy: the effective group and the ineffective group. The expression levels of nuclear factor-kappa B (NF-κB), matrix metalloproteinase 9 (MMP9), Ki-67, B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X (Bax), Vimentin, E-cadherin, and N-cadherin in tumor tissues were evaluated using reverse transcription-polymerase chain reaction (RT-PCR). In vitro, Huh7 cells were cultured, and lentivirus infections were utilized to inhibit the overexpression of NF-κB and MMP9. The determination of EMT and cell viability was conducted through Cell Counting Kit-8 (CCK-8) assays, RT-PCR, and Western blot. Sixty patients diagnosed with advanced liver cancer were selected for the study. Based on their clinical outcomes, 30 patients with advanced hepatocellular carcinoma were categorized into the effective group, while the remaining 30 patients were categorized into the ineffective group. The results of the Western blot analysis indicated that, in comparison to the effective group, the expression levels of NF-κB, MMP9, Ki-67, Bcl-2, Vimentin, and N-cadherin were significantly higher in the tumor tissues of the ineffective group. Conversely, the expression of Bax and E-cadherin was notably lower in the effective group. Following the individual knockdown of NF-κB and MMP9, the cell experiments revealed a remarkable decrease in the expression levels of Ki-67, Bcl-2, Vimentin, and N-cadherin, whereas the expression of Bax and E-cadherin showed significant elevation (p < 0.05). Furthermore, there was a significant increase in cell viability and a decrease in cell apoptosis after the knockdown of NF-κB and MMP9. The NF-κB/MMP9 signaling axis serves as a pivotal regulator that fosters proliferation and impedes apoptosis in Huh7 cells by modulating the process of EMT.

Irrigation water economic value and productivity: An econometric estimation for maize grain production in Italy

Climate change, characterized by rising temperatures and limited precipitation, has intensified the demand for irrigation water while simultaneously restricting its availability. This challenge poses significant risks to agricultural and food production, particularly in the Mediterranean regions where, recently, water deficits have led to substantial production losses and quality issues. Water is a critical determinant of crops' economic viability, especially for water-intensive crops, making it essential to estimate its economic relevance, especially in the absence of reliable water market prices. This study has two primary objectives: first, to evaluate the shadow price of irrigation water for maize grain at the farm level, which is defined as the value generated by the marginal unit of water consumed; and second, to analyse its heterogeneity. Leveraging a Farm Accountancy Data Network (FADN) panel of 1625 Italian farms over a decade (2010–2020), an econometric production function approach is employed. Moreover, quantile regressions reveal variations in the shadow price linked to geographical, managerial, and structural farm characteristics. Our findings underscore water’s key role in economically viable maize grain production, significantly enhancing the productivity of other inputs like fertilizers and pesticides. The average shadow price is 0.29 €/m³, with a median of 0.20 €/m³ and water total productivity accounts for one-third of maize’s average gross output. Quantile regressions uncover how factors like geographic location, altitude, farm management, irrigation water source, and farm size influence the distribution of water productivity, reflecting either efficient use or scarcity of this resource. Our estimation provides valuable insights for policymakers by offering accurate shadow price estimates for irrigation water in Italian maize grain production. Furthermore, it enhances our understanding of irrigation water’s role in the economic viability of this crop, while contributing to support evidence-based water management strategies, identifying vulnerable areas and farms and allowing for future methodological developments.

Cisplatin and Starvation Differently Sensitize Autophagy in Renal Carcinoma: A Potential Therapeutic Pathway to Target Variegated Drugs Resistant Cancerous Cells.

Cisplatin, a powerful chemotherapy medication, has long been a cornerstone in the fight against cancer due to chemotherapeutic failure. The mechanism of cisplatin resistance/failure is a multifaceted and complex issue that consists mainly of apoptosis inhibition through autophagy sensitization. Currently, researchers are exploring ways to regulate autophagy in order to tip the balance in favor of effective chemotherapy. Based on this notion, the current study primarily identifies the differentially expressed genes (DEGs) in cisplatin-treated autophagic ACHN cells through the Illumina Hi-seq platform. A protein-protein interaction network was constructed using the STRING database and KEGG. GO classifiers were implicated to identify genes and their participating biological pathways. ClueGO, David, and MCODE detected ontological enrichment and sub-networking. The network topology was further examined using 12 different algorithms to identify top-ranked hub genes through the Cytoscape plugin Cytohubba to identify potential targets, which established profound drug efficacy under an autophagic environment. Considerable upregulation of genes related to autophagy and apoptosis suggests that autophagy boosts cisplatin efficacy in malignant ACHN cells with minimal harm to normal HEK-293 growth. Furthermore, the determination of cellular viability and apoptosis by AnnexinV/FITC-PI assay corroborates with in silico data, indicating the reliability of the bioinformatics method followed by qRT-PCR. Altogether, our data provide a clear molecular insight into drug efficacy under starved conditions to improve chemotherapy and will likely prompt more clinical trials on this aspect.

Evaluating the Protective Effects of Thymoquinone on Methamphetamine-induced Toxicity in an In Vitro Model Based on Differentiated PC12 Cells.

Methamphetamine (Meth) is a highly addictive stimulant. Its potential neurotoxic effects are mediated through various mechanisms, including oxidative stress and the initiation of the apoptotic process. Thymoquinone (TQ), obtained from Nigella sativa seed oil, has extensive antioxidant and anti-apoptotic properties. This study aimed to investigate the potential protective effects of TQ against Meth-induced toxicity by using an in vitro model based on nerve growth factor-differentiated PC12 cells. Cell differentiation was assessed by detecting the presence of a neuronal marker with flow cytometry. The effects of Meth exposure were evaluated in the in vitro neuronal cell-based model via the determination of cell viability (in an MTT assay) and apoptosis (by annexin/propidium iodide staining). The generation of reactive oxygen species (ROS), as well as the levels of glutathione (GSH) and dopamine, were also determined. The model was used to determine the protective effects of 0.5, 1 and 2 μM TQ against Meth-induced toxicity (at 1 mM). The results showed that TQ reduced Meth-induced neurotoxicity, possibly through the inhibition of ROS generation and apoptosis, and by helping to maintain GSH and dopamine levels. Thus, the impact of TQ treatment on Meth-induced neurotoxicity could warrant further investigation.

Predicting forward osmosis performance with synthesized polyamide-based membrane: An integrated machine learning (MATLAB and ANN) and economic analysis framework

Forward Osmosis (FO) has emerged as a highly promising membrane-based water treatment process owing to its intrinsic advantages, such as low energy requirements and the potential for high water recovery. The lab-prepared highly efficient thin film composite (TFC) FO membranes that were used in this research work. The resultant polyamide-based FO membrane showed exceptional performance, notably achieving a high-water flux of 60.94 ± 3.5 L/m2.h and a constrained solute flux of 1.52 ± 0.08 g/m2.h. Machine learning methodologies are strategically employed in this study to construct predictive models for FO membrane performance. Notably, the accuracy of the van't Hoff equation's linear assumption is significantly enhanced by introducing an osmotic pressure calculation based on water activity, considering the nuanced effects of external and internal concentration polarization. The research employs MATLAB software alongside an artificial neural network (ANN) to develop predictive models. MATLAB facilitates the creation of a comprehensive theoretical model, offering insights into forecasting water flux through FO membranes by systematically analyzing diverse membrane parameters and their impact on performance. Concurrently, the ANN model was employed to predict the permeate flux for the laboratory-scale FO membrane system, utilizing the input parameters within the network's training range (R2 > 0.91). An economic assessment is also conducted to estimate the projected membrane cost, a pivotal determinant of system viability. Collectively, these investigations yield valuable insights into the intricacies of FO membrane design and optimization, elucidating their performance dynamics in water treatment and allied applications. This research significantly contributes to the ongoing quest to develop more efficient and precise membrane processes.