Decline In Viability Research Articles

A lef5-deficient baculovirus expression system with no virion contamination and promoting secretion

The baculovirus expression system (BEVS) is widely used in biomanufacturing. However, massive late and very late genes are expressed during baculovirus infection of the host cell, and the protein aggregation has a significant negative impact on the intercellular environment of cells and organelle function. Baculovirus particle contamination also hinders the purification of particulate products. In this study, a novel baculovirus vector of deficient-lef5 (Ac-Δlef5) was established to prevent the production of baculovirus particles. It could transduce cells and replicate viral DNA but the expression of the late and very late genes was reduced so that no virus was packed and budded. The Ac-Δlef5 expression system was successfully used to express a secreted protein, thyroid peroxidase (TPO). Compared to wild-type baculovirus (Ac-wt), Ac-Δlef5 delayed the decline in cell viability and prolonged the harvest period from 4 to 6 days after infection. When expressed in the Ac-Δlef5 system, 98% of TPO was secreted extracellularly, which was about 1.8 times that of the Ac-wt system. Meanwhile, the transcription levels of protein folding-related genes were significantly increased. The results show that the Ac-Δlef5 system is a potential novel viral-free baculovirus expression system, which omits the virion removal operation in biomanufacturing.

Quantitative Assessment of Oil Palm Pollen After 23 Years Of Cryobanking

Cryopreservation of pollen grains is an effective means of conserving desired germplasm of crop plants. Cryoconserved pollen are expected to be long-lived and thus can be suitably retrieved to overcome hybridization constraints imposed by a variety of reasons. We ascertained the performance of oil palm pollen grains (Tenera hybrids) that were cryobanked 23 years ago using liquid nitrogen (196°C). Cryostored pollen were assessed for viability, in-vitro germinability and vigour. Our analysis showed a marginal decline in viability, assessed through fluorochromatic reaction test, of cryopreserved pollen as compared to fresh ones (pre-storage assessment); however, the viability did not decline in the cryostate since it was last tested 15 years back. On the other hand, germinability and vigour of cryopreserved pollen were maintained to the levels of fresh pollen. Our study, for the first time, demonstrates the amenability of pollen grains for cryopreservation of any plant species beyond a period of two decades in general, and that for oil palm in particular.

Are orthodox Australian rainforest seeds short-lived in storage?

Context Fifty per cent of Australian rainforest species produce orthodox seeds, but little is known about their longevity in storage. Aims To (1) estimate the longevity of seeds of 33 rainforest species using artificial aging; (2) assess the influence of habit, habitat range (restricted to rainforest or more broadly distributed), seed characteristics (endospermy, dry weight), and collection environment (e.g. precipitation, elevation and average temperature) on longevity; (3) compare longevity of rainforest seeds to previously assessed species from non-rainforest habitats; (4) compare longevity in artificial aging to real-time longevity in storage. Methods Seeds were aged at 60% relative humidity and 45°C, and tested at pre-determined intervals until germination was reduced to zero. The time taken for germination to decline to 50% (p50AA) was calculated by probit analysis. Ordinary least-squares regression was used to model p50AA for rainforest and non-rainforest species against predictor variables. Values for p50AA were then compared with the actual longevity in storage at −20°C (p50RT) using Pearson’s correlation. Key results Species restricted to rainforest exhibited a significantly lower p50AA than species with a wider distribution or those restricted to non-rainforest habitats. Collection elevation had a significant negative influence on p50AA. In all, 14 of 33 species showed a significant decline in viability after ≤12 years in storage. Values of p50AA were not correlated with p50RT. Conclusions Rainforests contain a high proportion of potentially, and actually, short-lived species; however, p50AA is not a good predictor of their real-time longevity. Implications Rainforest species should be managed as short-lived, with viability checked at least every 5 years, until real-time longevity data indicate otherwise.

The adverse and beneficial effects of polyphenols in green and black teas in vitro and in vivo.

Although numerous studies highlight the health benefits of tea, excessive consumption has been linked to toxic conditions. Thus, understanding the optimal consumption of tea is essential to minimize toxicity while maximizing its benefits. In this study, we investigated the effects of eight green tea samples (G1-G8) and eight black tea samples (R1-R8) from Camellia sinensis, the most popular teas in Asian culture, on RSC96 Schwann neural cells and embryonic cardiomyocyte H9c2 cells. The results showed that the IC50 (mg/ml, weight/volume) of both tea types were inversely proportional to their polyphenol content, suggesting a relationship between toxicity and polyphenol levels in both green and black tea. Interestingly, green teas generally have higher polyphenol content than black teas. We also assessed the protective effects of tea in vitro by pretreating cells with the teas at indicated doses of polyphenol and subsequently exposing them to H2O2. Both tea types significantly reduced the decline in cell viability for both cell lines, and there was no significant difference in protective polyphenol concentrations for green (G3 & G7) and black (R3 & R8) teas at effective concentrations (EC20 and EC40). To evaluate the preventative effects of tea in vivo, we examined the impact of two green (G3 & G7) and two black (R3 & R8) teas with varying polyphenol content on dextran sulfate sodium (DSS)-induced inflammatory colitis in mice. Tea-treated groups exhibited significantly lower inflammatory scores (DAI) than the control group. DSS treatment in the control group led to shortened colorectal lengths in mice, while tea co-treatment partially prevented this loss. Histological analysis revealed that G7 and R3 (with a moderate polyphenol content) treatment improved colorectal crypt structure, decreased the severity of inflammatory ulcerative colitis, and significantly reduced histological scores compared to the control group. However, G3 and R8 (with high and low doses of polyphenol content, respectively) did not show these effects, suggesting that a moderate polyphenol level in both tea types is optimal for preventative benefits.

Dietary cobalt oxide nanoparticles alleviate aging through activation of mitochondrial UPR in Caenorhabditis elegans.

Mitochondrial unfolded protein response (UPRmt), which is a mitochondrial proteostasis pathway, orchestrates an adaptive reprogramming for metabolism homeostasis and organismal longevity. Similar to other defense systems, compromised UPRmt is a feature of several age-related diseases. Here we report that dimercapto succinic acid (DMSA)-modified cobalt oxide nanoparticles (Co3O4 NPs), which have received wide-spread attention in biomedical fields, is a promising UPRmt activator and, more importantly, provides a gate for extending healthy lifespan. Methods: UPRmt activation by Co3O4 NPs was tested in transgenetic Caenorhabditis elegans (C. elegans) specifically expressing UPRmt reporter Phsp-6::GFP, and the underlying mechanism was further validated by mitochondrial morphology, mtDNA/nDNA, metabolism-related genes' expression, mitonuclear protein imbalance, oyxgen assumption and ATP level in C. elegans. Then therapeutic response aganist senescence was monitored by lifespan analysis, lipofusin contents, MDA contents, Fe accumulation, pharyngeal locomotion performance as well as athletic ability (head thrashes and body bends) at different developmental stages of C. elegans. RNAi towards ubl-5 or atfs-1 in UPRmt pathway was applied to clarify the role of UPRmt in Co3O4 NPs -mediated anti-aging effects. Finally, the effect of Co3O4 NPs on mitochondrial homeostasis and D-galactose-induced cell viability decline in mammalian cells were studied. Results: Co3O4 NPs was revealed as a bona fide activator of the UPRmt signaling pathway, through fine-tuning mitochondrial dynamics and inducing a stoichiometric imbalance between OXPHOS subunits encoded by mitochondrial DNA (mtDNA) and nuclear DNA (nDNA) at early life stage of C. elegans. Phenotypically, Co3O4 NPs treatment protect C. elegans from external stresses. More importantly, dietary low level of Co3O4 NPs effectively extend lifespan and alleviate aging-related physiological and functional decline of worms, demonstrating its potential roles in delaying aging. While the protective effect exerted by Co3O4 NPs was compromised in line with atfs-1 or ubl-5 RNAi treatment. Further studies verified the conservation of Co3O4 NPs in activating UPRmt and exerting protective effects in mammalian cells. Conclusions: The results reveal beneficial effects of Co3O4 NPs on mitochondrial metabolic control, thus presenting their potential efficacy in anti-aging care.

Determinants of Farm Revenue in Pakistan

Will small farm viability decline with the reduction of average farm size in Pakistan? This paper addresses the determinants of rural household and farm-related income. Using the 2001 PIDE Household Survey, the approach developed captures the potential interactions between farm returns and household, farm, and factor market characteristics (schooling, family size, land tenure and operational size, access to water, credit, and capital). Econometric results show: (a) returns to additional schooling and the revenue elasticity of operated acres increase with farm size; (b) medium and large farm renters would be willing to pay more than observed rents, implying an incentive to increase farm size at the prevailing rental values; (c) owneroperated farms, landowners who also leases in, and fixed rental tenants earn higher revenues than sharecropping tenants. The difference, however, between landowner/fix-renter income and sharecropper income varies with family and farm size, as well as water use. While these results favour farm size increase, the results also show that off-farm and non-farm income sources are relatively more important for small farmers, contributing to their viability. JEL classification: D13, Q12, Q15 Keywords: Pakistan, Land Markets, Rural Factor Markets, Revenue Function

Polymyxin B neutralizes endotoxin and improves the quality of chicken semen during liquid storage

Despite its importance in gamete utilization for livestock production, poultry semen cryopreservation in a liquid state, is limited in the poultry industry due to a significant decline in sperm viability and functionality during liquid storage. Lipopolysaccharide (LPS) is released from gram-negative bacteria and impairs sperm function in mammals. Using exogeneous LPS, we show this endotoxin compromises sperm viability and function, including motility and penetrability to the inner peri-vitelline layer (IPVL) during liquid storage at 4 °C. This outcome was supported by LPS quantification showing an extreme increase in the first 24 h of storage. Polymyxin B (PMB) is an LPS neutralizer previously shown to improve fertility in boar semen, thus we explored the effect of PMB on chicken semen quality during liquid storage. Sperm viability and penetrability tests showed that PMB completely abolishes the deleterious effect by LPS. However, co-addition of PMB with penicillin G (PenG), an antibiotic against gram positive bacteria, reduces IPVL-penetrability while improving sperm viability post-storage. Furthermore, artificial insemination trials showed that PMB addition improves semen fertility at the post liquid storage. Our results show that chicken semen quality during liquid storage is impaired by bacterial LPS, but improved by PMB addition due to cancelled endotoxic effects, which offers a new approach for prolonged fertility of poultry semen storage in a liquid state.

Istradefylline modulates purinergic enzymes and reduces malignancy-associated factors in B16F10 melanoma cells.

ATP and adenosine exert pivotal roles in the development, maintenance, and metastatic spreading of melanoma. The action of such key melanoma tumor microenvironment (TME) constituents might be complementary or opposed, and their effects are not exclusive to immune cells but also to other host cells and tumor cells. The effects of ATP are controlled by the axis CD39/73, resulting in adenosine, the main actor in the TME, and A2A is the crucial mediator of its effects. We evaluated ATP and adenosine signaling through A2A on B16F10 melanoma cells using istradefylline (IST) (antiparkinsonian A2A antagonist) and caffeine (CAF) treatments after exposure to ATP and adenosine. Adenosine increased melanoma cell viability and proliferation in a concentration-dependent manner. ATP increases viability only as a substrate by CD39 to produce adenosine. Both IST and CAF are toxic to B16F10 cells, but only IST potentialized paclitaxel-induced cytotoxic effects, even decreasing its IC50 value. IST positively modulated CD39 and CD73 expression. CD39 activity was increased, and E-ADA was reduced, indicating that the melanoma cells promoted compensatory feedback in the production and maintenance of adenosine levels. A2A antagonism by IST reduced the factors associated with malignancy, like migration, adhesion, colony formation, and the capacity to produce melanin. Moreover, IST significantly increases nitric oxide (NO) production, which correlates to a decline in melanoma cell viability by apoptotic events. Altogether, our results suggest that adenosine signaling through A2A is essential for B16F10 cells, and its inhibition by IST causes compensatory purinergic enzymatic modulations. Furthermore, IST is a promising therapy that provides new ways to improve current melanoma treatments.

AP2a-Mediated Upregulation of miR-125a-5p Ameliorates Radiation-Induced Oxidative Stress Injury via BRD4/Nrf2/HO-1 Signaling.

Radiation therapy is widely used to restrain tumor progression, but it is always accompanied by damage to healthy tissues. We aimed to probe the impact and mechanism of activator protein 2a (AP2a) and miR-125a-5p in radiation-induced oxidative stress injury. Human umbilical vein endothelial cells (HUVECs) were treated with X rays to induce radiation injury in vitro. Cell viability was measured using MTT assays. Flow cytometry assay was employed to detect the apoptosis rate. Oxidative stress markers were evaluated by detection kits. Gene or protein levels were determined by RT-qPCR or Western blotting. Validation of the interaction of miR-125a-5p with BRD4 and AP2a was conducted by dual luciferase assay or ChIP. MiR-125a-5p and AP2a were decreased in irradiated HUVECs, whereas BRD4 was increased. MiR-125a-5p overexpression or BRD4 silencing alleviated the cell viability decline, apoptosis, and oxidative stress injury caused by radiation treatment. MiR-125a-5p repressed the BRD4 level. The protective effects of miR-125a-5p overexpression in the radiation-induced oxidative injury were impeded by BRD4 overexpression. Moreover, AP2a bound to the promoter of miR-125a-5p. MiR-125a-5p inhibition reversed the effects of AP2a overexpression on radiational oxidative injury by modulating Nrf2/HO-1 signaling. AP2a transcriptionally activated miR-125a-5p ameliorated oxidative stress injury of HUVECs caused by radiation through Nrf2/HO-1 signaling.

Non-human animal cultures, co-cultures and conservation

Abstract Animal culture involves the transmission of behaviours through social learning, which plays a crucial role in species’ survival and adaptability to changing environments. Co-cultures, which describe the mutual cultural evolution between species, underscore the importance of interspecies interactions in maintaining ecological balance and resilience. The loss of these cultural practices can lead to a decline in population viability, reduced genetic diversity, and destabilised ecosystems, ultimately impacting the services these ecosystems provide to human health and well-being. The preservation of cultural traits is essential for maintaining biodiversity, ensuring species’ adaptive capacities, and supporting the overall health of ecosystems, which are critical for the survival of both non-human animals and humans. Integrating the conservation of animal cultures and co-cultures into broader conservation and One Health strategies is necessary to protect the delicate balance of life on Earth and ensure the well-being of all species.

The Effect of Bacteria on the Stability of Microfluidic-Generated Water-in-Oil Droplet.

Microencapsulation in emulsion droplets has great potential for various applications such as food which require formation of highly stable emulsions. Bacterial-emulsion interactions affect the physiological status of bacteria while bacterial cell characteristics such as surface-active properties and metabolic activity can affect emulsion stability. In this study, the viability and growth of two different bacterial species, Gram-negative Escherichia coli and Gram-positive Lactobacillus paracasei, encapsulated in water-in-oil (W/O) droplets or as planktonic cells, were monitored and their effect on droplet stability was determined. Microencapsulation of bacteria in W/O droplets with growth media or water was achieved by using a flow-focusing microfluidic device to ensure the production of highly monodispersed droplets. Stability of W/O droplets was monitored during 5 days of storage. Fluorescence microscopy was used to observe bacterial growth behaviour. Encapsulated cells showed different growth to planktonic cells. Encapsulated E. coli grew faster initially followed by a decline in viability while encapsulated L. paracasei showed a slow gradual growth throughout storage. The presence of bacteria increased droplet stability and a higher number of dead cells was found to provide better stability due to high affinity towards the interface. The stability of the droplets is also species dependent, with E. coli providing better stability as compared to Lactobacillus paracasei.

Atorvastatin Inhibits Ferroptosis of H9C2 Cells by regulatingSMAD7/Hepcidin Expression to Improve Ischemia-Reperfusion Injury.

Ferroptosis plays a key role in cardiomyopathy. Atorvastatin (ATV) has a protective effect on ischemia-reperfusion (I/R) cardiomyopathy. The purpose of this study is to elucidate the mechanism of ATV in I/R injury. H9C2 cells and cardiomyopathy rats were induced by hypoxia/reoxygenation (H/R) and I/R to construct in vitro and in vivo models. Cell viability was determined by CCK8. Cardiac histopathology was observed by HE staining. Transmission electron microscope (TEM) was used to observe the mitochondrial morphology. The reactive oxygen species (ROS) content in cells was analyzed by the biochemical method. ELISA was conducted to calculate the concentrations of total iron/Fe2+ and hepcidin. The expression of ferroptosis and SMAD pathway-related genes were detected by qPCR. Western blot was performed to detect the expression levels of ferroptosis and SMAD pathway-related proteins. In H9C2 cells, ATV reversed the decline in cell viability, mitochondrial shrinkage, and ROS elevation induced by erastin or H/R. The concentration of total iron and Fe2+ in H/R-induced H9C2 cells increased, and the protein expression of FPN1 decreased. After ATV treatment, the concentration of total iron and Fe2+ decreased, and the protein expression of FPN1 increased. The expression of the SMAD7 gene in H/R-induced H9C2 cells decreased, and the expression of the hepcidin gene increased, which were reversed by ATV. When SMAD7 was knocked down, ATV treatment failed to produce the above effect. ATV also improved ferroptosis in I/R rat myocardium through the SMAD7/hepcidin pathway. ATV reversed the decline in H9C2 cell viability, mitochondrial shrinkage, and ROS elevation, and improved the myocardium ferroptosis through the SMAD7/hepcidin pathway in I/R rat.

Spermidine inhibits high glucose-induced endoplasmic reticulum stress in HT22 cells by upregulation of growth differentiation factor 11.

Hyperglycemia-induced neuronal endoplasmic reticulum (ER) stress is particularly important for the pathogenesis of diabetic encephalopathy. Spermidine (Spd) has neuroprotection in several nervous system diseases. Our current study to explore the potential protective role of Spd in hyperglycemia-induced neuronal ER stress and the underlying mechanisms. HT22 cells were treated with high glucose (HG) to establish an in-vitro model of hyperglycemia toxicity. The HT22 cells' activity was tested by cell counting kit-8 assay. RNA interference technology was used to silence the expression of growth differentiation factor 11 (GDF11) in HT22 cells. The GDF11 expression levels of mRNA were assessed using reverse transcription-PCR (RT-PCR). Western blotting analysis was applied to evaluate the expressions of GRP78 and cleaved caspase-12. Spd markedly abolished HG-exerted decline in cell viability as well as upregulations of GRP78 and cleaved caspase-12 in HT22 cells, indicating the protection of Spd against HG-induced neurotoxicity and ER stress. Furthermore, we showed that Spd upregulated the expression of GDF11 in HG-exposed HT22 cells. While, silenced GDF11 expression by RNA interference reversed the protective effects of Spd on HG-elicited neurotoxicity and ER stress in HT22 cells. These results indicated that Spd prevents HG-induced neurotoxicity and ER stress through upregulation of GDF11. Our findings identify Spd as a potential treatment for diabetic encephalopathy as well as ER stress-related neurologic diseases.

Interactive effect between tree ageing and trunk-boring pest reduces hydraulics and carbon metabolism in Hippophae rhamnoides.

Sea-buckthorn (Hippophae rhamnoides) is widely distributed across the Eurasian continent. Recently sea-buckthorn has shown premature ageing and decline when confronted with water deficiency and Holcocerus hippophaecolus damage in northwest China and the Loess Plateau region. However, the physiological process of sea-buckthorn senescence in response to drought and pest damage is still unknown. In this study, 4-year-old (4y), 15-year-old normal growth (15yN) and 15-year-old seriously moth-damaged sea-buckthorn plants (15yH) were used as the research objects. The growth of branches and roots, branch water potential and percentage loss of conductivity (PLC), branch vulnerability to embolism (quantified by P50, xylem water potential at 50 % of PLC), branch xylem parenchyma cell viability, photosynthesis and the non-structural carbohydrate (NSC) content in branches and roots in dry and wet seasons were measured. The results showed that the length, basal diameter of 1-year-old branches and the leaf area of 4y trees were significantly larger than that of 15yN and 15yH trees, and the fine root density of 15yH trees was significantly lower than that of 15yN trees in all measured areas. The branch-specific hydraulic conductivity of 15yN and 15yH trees was only 50.2 % and 12.3 % of that of 4y trees, and the P50 of 4y, 15yH and 15yN trees was -3.69 MPa, -2.71 MPa and -1.15 MPa, respectively. The midday water potential and photosynthetic rate were highest in 4y trees, followed by 15yN and then 15yH trees in both the dry season and wet seasons, while branch PLC declined in the opposite direction (15yH trees highest, 4y trees lowest). The degree of PLC repair within a day was highest in 4y trees, followed by 15yN and then 15yH trees, and the viability of xylem cells was consistent with this pattern. The branch xylem starch and NSC content of 4y and 15yN trees were significantly higher than that of 15yH trees in the dry season, and the root starch and NSC content of 4y trees were significantly higher than that of 15yH trees in the two seasons. The above results suggest that the hydraulic properties of the normal elderly and seriously pest-damaged sea-buckthorn were significantly worse than in juvenile plants. Narrower early wood width and vessel density, high embolism vulnerability and weak embolism repair capacity led to the decline in water-conducting ability, and similarly further affected photosynthesis and the root NSC content. The decline in xylem parenchyma cell viability was the main reason for the limited embolism repair in the branches.

Adiponectin Protects Hypoxia/Reoxygenation-Induced Cardiomyocyte Injury by Suppressing Autophagy.

Adiponectin is a cytokine produced by adipocytes and acts as a potential cardioprotective agent and plays an important role in myocardial ischemia/reperfusion injury. In a myocardial hypoxia/reoxygenation model using neonatal rat ventricular myocytes, we investigated the contribution of adiponectin-mediated autophagy to its cardioprotective effects. Cardiomyocytes were exposed to hypoxia/reoxygenation pretreated with or without adiponectin in the presence of absence of rapamycin. Cell viability was analyzed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. Western blotting assay was used to determine the expression levels of microtubule-associated proteins 1A/1B light chain 3B (LC3B), adenosine monophosphate-activated protein kinase (AMPK), mammalian target of rapamycin (mTOR), p62/sequestosome 1, unc-51 like autophagy activating kinase 1 (ULK1), and Beclin-1. Autophagosome formation was detected by monodansylcadaverine staining. We found that hypoxia induced a time dependent decline in cardiomyocyte viability, and increase in autophagy and reoxygenation further augmented hypoxia-induced autophagy induction and consequently reduced cell viability. Adiponectin treatment alleviated hypoxia/reoxygenation-induced cellular damage and autophagy in cardiomyocytes. Adiponectin treatment also attenuated hypoxia/reoxygenation-promoted cardiomyocyte autophagy even in the presence of another autophagy stimulator rapamycin in part by inhibiting vacuolar hydron-adenosine triphosphatase. Additionally, autophagy suppression by adiponectin during hypoxia/reoxygenation was associated with the attenuated phosphorylation of AMPK and ULK1, augmented phosphorylation of mTOR, and the reduced protein expression levels of Beclin-1 in cardiomyocytes. Taken together, these results suggest that adiponectin protects ischemia/reperfusion-induced cardiomyocytes by suppressing autophagy in part through AMPK/mTOR/ULK1/Beclin-1 signaling pathway.

Synthesis of M-Ag3PO4, (M = Se, Ag, Ta) Nanoparticles and Their Antibacterial and Cytotoxicity Study.

Silver Phosphate, Ag3PO4, being a highly capable clinical molecule, an ultrasonic method was employed to synthesize the M-Ag3PO4, (M = Se, Ag, Ta) nanoparticles which were evaluated for antibacterial and cytotoxicity activities post-characterization. Escherichia coli and Staphylococcus aureus were used for antibacterial testing and the effects of sonication on bacterial growth with sub-MIC values of M-Ag3PO4 nanoparticles were examined. The effect of M-Ag3PO4 nanoparticles on human colorectal carcinoma cells (HCT-116) and human cervical carcinoma cells (HeLa cells) was examined by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) assay and DAPI (4′,6-diamidino-2-phenylindole) staining. Additionally, we analyzed the effect of nanoparticles on normal and non-cancerous human embryonic kidney cells (HEK-293). Ag-Ag3PO4 exhibited enhanced antibacterial activity followed by Ta-Ag3PO4, Ag3PO4, and Se-Ag3PO4 nanoparticles against E. coli. Whereas the order of antibacterial activity against Staphylococcus aureus was Ag3PO4 > Ag-Ag3PO4 > Ta-Ag3PO4 > Se-Ag3PO4, respectively. Percentage inhibition of E. coli was 98.27, 74.38, 100, and 94.2%, while percentage inhibition of S. aureus was 25.53, 80.28, 99.36, and 20.22% after treatment with Ag3PO4, Se-Ag3PO4, Ag-Ag3PO4, and Ta-Ag3PO4, respectively. The MTT assay shows a significant decline in the cell viability after treating with M-Ag3PO4 nanoparticles. The IC50 values for Ag3PO4, Se-Ag3PO4, Ag-Ag3PO4, and Ta-Ag3PO4 on HCT-116 were 39.44, 28.33, 60.24, 58.34 µg/mL; whereas for HeLa cells, they were 65.25, 61.27, 75.52, 72.82 µg/mL, respectively. M-Ag3PO4 nanoparticles did not inhibit HEK-293 cells. Apoptotic assay revealed that the numbers of DAPI stained cells were significantly lower in the M-Ag3PO4-treated cells versus control.

Probing the Potential Mechanism of Quercetin and Kaempferol against Heat Stress-Induced Sertoli Cell Injury: Through Integrating Network Pharmacology and Experimental Validation.

Quercetin and kaempferol are flavonoids widely present in fruits, vegetables, and medicinal plants. They have attracted much attention due to their antioxidant, anti-inflammatory, anticancer, antibacterial, and neuroprotective properties. As the guarantee cells in direct contact with germ cells, Sertoli cells exert the role of support, nutrition, and protection in spermatogenesis. In the current study, network pharmacology was used to explore the targets and signaling pathways of quercetin and kaempferol in treating spermatogenic disorders. In vitro experiments were integrated to verify the results of quercetin and kaempferol against heat stress-induced Sertoli cell injury. The online platform was used to analyze the GO biological pathway and KEGG pathway. The results of the network pharmacology showed that quercetin and kaempferol intervention in spermatogenesis disorders were mostly targeting the oxidative response to oxidative stress, the ROS metabolic process and the NFκB pathway. The results of the cell experiment showed that Quercetin and kaempferol can prevent the decline of cell viability induced by heat stress, reduce the expression levels of HSP70 and ROS in Sertoli cells, reduce p-NF-κB-p65 and p-IκB levels, up-regulate the expression of occludin, vimentin and F-actin in Sertoli cells, and protect cell structure. Our research is the first to demonstrate that quercetin and kaempferol may exert effects in resisting the injury of cell viability and structure under heat stress.

Effects of urea formaldehyde resin on the characteristics and microbial community of anaerobic granular sludge

The effects of urea formaldehyde (UF) resin on the performance and characteristics of anaerobic granular sludge (AnGS), the microbial community and related inhibition mechanisms were studied. During the exposure period, a low concentration of UF resin (0.1 g/L) had no significant effect on the AnGS activity, while a high concentration of UF resin (0.2 g/L, 0.5 g/L) decreased methane production and chemical oxygen demand (COD) removal by 16.5–19.7% and 6.4–7.9%, and short-chain fatty acid (SCFA) increased by 17.1–18.9%. Meanwhile, a low concentration of UF resin could stimulate cell stress, induce the production of more EPS and realize the self-defence mechanism; however, a higher UF resin concentration will inhibit the production of EPS, the extracellular polymer in the reactor with high concentration of UF resin was only 83.1–69.3% of that in control group, it leads to the crushing of AnGS and the decline of cell viability. In addition, high concentration of UF resin reduced the relative abundance of Proteobacteria and Bacteroidota by 9.2%− 16.7% and 8.0%− 10.5% respectively, led to the decline of bacterial diversity and richness, and the metabolic performance in anaerobic system was prepared. UF resin induced oxidative stress has been proved to be the main reason for the decline of AnGS activity, this is reflected in the increase of reactive oxygen species (ROS) production and lactate dehydrogenase (LDH) release, this further explains the reason why UF resin inhibits sludge acidification and methanation.

MiR-702-5p ameliorates diabetic encephalopathy in db/db mice by regulating 12/15-LOX

The purpose of this study was to investigate the effect of miR-702-5p on diabetic encephalopathy (DE) and the interaction of miR-702-5p/12/15-LOX in the central nervous system (CNS). In this study, db/db mice were used as DE animal model and HT22 cells were treated with high-glucose (HG). Based on the bioinformatics prediction of possible binding sites between miR-702-5p and 12/15-LOX, we found that the expression of miR-702-5p was significantly down-regulated while 12/15-LOX up-regulated in vivo and in vitro, and the expression changes were inversely correlated. In vivo, diabetic mice with cognitive dysfunction and hippocampal neuronal damage had a concomitant increase in amyloid precursor protein (APP), amyloid beta(Aβ), tau, BAX protein expressions; by contrast, Bcl-2 protein expression was significantly decreased. Overexpression of miR-702-5p significantly reduced the histopathological damage of the hippocampus, improved the learning and memory function of db/db mice, down-regulated 12/15-LOX, APP, Aβ, tau, BAX protein expressions significantly and up-regulated the expression of Bcl-2. In vitro, miR-702-5p mimic reversed the decline in cell viability and the increase in cell apoptosis induced by HG. Simultaneously, reduced 12/15-LOX, APP, Aβ, BAX protein expressions, and increased Bcl-2 protein expression were detected in the miR-702-5p mimic group. Moreover, combined administration of miR-702-5p mimic and 12/15-LOX overexpression lentivirus significantly reversed the protective effect of up-regulation of miR-702-5p. In conclusion, miR-702-5p has a neuroprotective effect on DE, and this effect was achieved by inhibiting 12/15-LOX. However, miR-702-5p had an endogenous regulatory effect on 12/15-LOX rather than a direct targeting relationship.

Melatonin inhibits NaIO3-induced ARPE-19 cell apoptosis via suppression of HIF-1α/BNIP3-LC3B/mitophagy signaling

BackgroundAge-related macular degeneration (AMD) leads to gradual central vision loss and eventual irreversible blindness. Melatonin, an endogenous hormone, exhibits anti-inflammatory and antitumor effects; however, the role it plays in AMD remains unclear. Herein, we investigated the anti-AMD molecular mechanism of melatonin after sodium iodate (NaIO3) treatment of ARPE-19 cells in vitro and in animal models with the goal of improving the therapeutic effect.ResultsThe in vitro results showed that melatonin protected against NaIO3-induced cell viability decline, mitochondrial dysfunction and apoptosis in ARPE-19 cells, and melatonin also alleviated NaIO3-induced reactive oxygen species (ROS) production, mitochondrial dysfunction and mitophagy activation. Melatonin reduced NaIO3-induced mitophagy activation through HIF-1α-targeted BNIP3/LC3B transcription, whereas ROS inhibition realized with N-acetylcysteine (NAC, a ROS inhibitor) combined with melatonin reduced the effect of NaIO3 on mitophagy. An animal model of AMD was established to confirm the in vitro data. Mouse tail vein injection of NaIO3 and melatonin was associated with enhanced repair of retinal layers within 7 days, as observed by optical coherence tomography (OCT) and hematoxylin and eosin (H&E) staining. A reduction in BNIP3 and HIF-1α levels, as determined by immunohistochemistry (IHC) assay, was also observed.ConclusionsThese results indicate that melatonin attenuated NaIO3-induced mitophagy of ARPE-19 cells via reduction in ROS-mediated HIF-1α targeted BNIP3/LC3B signaling in vitro and in vivo. Melatonin may be a potential therapeutic drug in the treatment of AMD.