Dose-dependent Rise Research Articles

"Effect of Sunset Yellow on Testis: Molecular Evaluation, and Protective Role of Coenzyme Q10 in Male Sprague-Dawley Rats".

In recent years, Sunset Yellow (SY) has been widely used as a food additive, sparking debates about its potential toxicity. This research aims to investigate SY's effects at both the molecular and histopathological levels, along with the protective benefits of Coenzyme Q10 (CoQ10) supplementation in male rat testes. Forty-two male Sprague-Dawley rats were randomly divided into six groups (n = 7) and given daily oral gavages for six weeks. The groups included: a low dose of Sunset Yellow (2.5 mg/kg/day), a high dose of Sunset Yellow (70 mg/kg/day), CoQ10 (10 mg/kg/day), CoQ10 with the low dose of Sunset Yellow, CoQ10 with the high dose of Sunset Yellow, and deionized water as a control. After anesthesia, the rats' testes were removed for molecular and histological analysis. The findings showed a dose-dependent rise in the expression of oxidative stress genes (Sod, Gpx, and Cata) and a notable decrease in the expression of the steroidogenic acute regulatory (Star) gene (P value < 0.05) with increasing SY doses. Histological results supported these outcomes. Additionally, there was no significant distinction between rats treated with CoQ10 along with low doses of Sunset Yellow (CoQ10+LD) and control rats given low doses of Sunset Yellow (SY-LD). Conclusions: This study illustrates that SY, as an artificial food dye, has harmful effects on the male reproductive system, while the utilization of CoQ10 can alleviate the negative impacts of SY exposure.

The influence of ghrelin agonist ipamorelin acetate on the hypothalamic-pituitary-testicular axis in a cichlid fish, Oreochromis mossambicus

Ghrelin, a peptide found in the brain and gut, is predicted to play a significant role in the control of various physiological systems in fish. The objective of this study was to examine the impact of ipamorelin acetate (IPA), a ghrelin agonist, on the reproductive axis of the tilapia Oreochromis mossambicus. The administration of either 5 or 30 µg of IPA for 21 days led to a significant and dose-dependent rise in food intake concomitant with a significant increase in the numbers of primary spermatocytes, secondary spermatocytes, and early spermatids compared to the control group. There was a significant rise in the number of late spermatids, as well as the areas of the lobule and lumen, in fish treated with 30 µg of IPA, compared to the control group. Moreover, there was no significant difference in the percentage of gonadotropin-releasing hormone (GnRH)-immunoreactive fibres in the hypothalamus and anterior pituitary gland across different groups. However, a significant elevation in the expression of androgen receptor protein was observed in fish treated with 30 µg of IPA. Furthermore, the concentrations of luteinizing hormone (LH) and 11-ketotestosterone (11-KT) in the serum of fish treated with either 5 or 30 µg of IPA were significantly elevated in comparison to the control group. Collectively, these findings suggest that the administration of ghrelin enhances the development of germ cells during the meiosis-I phase and that this effect might be mediated via the stimulation of 11-KT and androgen receptors at the testicular level and LH at the pituitary level in the tilapia.

Vallaris solanacea induces mitochondrial mediated apoptosis in HL-60 human promyelocytic leukemia cells

In the present study, the apoptosis-inducing potential of a chloroform fraction from an alcoholic extract of Vallaris solanacea aerial parts (VS) was examined using human promyelocytic leukemia HL-60 cells. We discovered a concentration and time-dependent decrease in cell growth using MTT assay. Scanning electron micrographs and fluorescence microscopy were used to observe several well-documented morphological and nuclear alterations, such as reduction in cell size, chromatin condensation, fragmentation, and the creation of cell surface blebs. A considerable rise in the Sub-G0 population was revealed by cell cycle analysis. Additionally, a dose-dependent rise in cells positive for Annexin V was observed. DCFH-DA test on VS-treated HL-60 cells showed an increase in endogenous ROS generation of up to 4.3 fold. Additionally, suppression in Bcl-2 levels and increased mitochondrial membrane depolarization in treated cells were also associated with a rise in cytosolic cytochrome-c levels that was consequently followed by the activation of the caspase cascade. Further, the DNA fragmentation assay exhibited a typical ladder formation at 25 μg/ml, which became prominent in a concentration-dependent manner. Our study revealed that VS has apoptosis-inducing potential towards HL-60 cells in vitro and is an effective candidate for further anti-cancer studies.

N-3 fatty acids and the risk of atrial fibrillation, review.

Atrial fibrillation (AF) is the most frequent type of cardiac arrhythmia that affects over six million individuals in Europe. The incidence and prevalence of AF rises with age, and often occurs after cardiac surgery. Other risk factors correlated with AF comprise high blood pressure, diabetes mellitus, left atrial enlargement, ischemic heart disease, and congestive heart failure. Considering the high prevalence of AF in aging societies, strategies to prevent serious complications, such as stroke or heart failure, are important because they are correlated with high morbidity and mortality. The supplementation of sea-derived n-3 polyunsaturated fatty acids (PUFA) is widely discussed in this context, but the results of experimental and observational studies are in contrast to randomized placebo-controlled intervention trials (RCTs). Specifically, larger placebo-controlled n-3 PUFA supplementation studies with long follow-up showed a dose-dependent rise in incident AF. Daily n-3 PUFA doses of≥1 g/d are correlated with a 50 % increase in AF risk, whereas a daily intake of <1 g/d causes AF in only 12 %. Individuals with a high cardiovascular risk (CVD) risk and high plasma-triglycerides seem particularly prone to develop AF upon n-3 PUFA supplementation. Therefore, we should exercise caution with n-3 PUFA supplementation especially in patients with higher age, CVD, hypertriglyceridemia or diabetes. In summary, existing data argue against the additive intake of n-3 PUFA for preventative purposes because of an incremental AF risk and lacking CVD benefits. However, more clinical studies are required to disentangle the discrepancy between n-3 PUFA RCTs and observational studies showing a lower CVD risk in individuals who regularly consume n-3 PUFA-rich fish.

Development of Syringaldehyde as an Agonist of the GLP-1 Receptor to Alleviate Diabetic Disorders in Animal Models.

The phenolic aldehyde syringaldehyde (SA) has been shown to have an antihyperglycemic effect in diabetic rats due to increased glucose utilization and insulin sensitivity. To understand the direct effect of SA on the GLP-1 receptor, STZ-induced diabetic rats were used. The levels of pro-inflammatory cytokines, liver enzymes, and renal function were measured using specific ELISA kits. The mechanisms of SA effects were investigated using CHO-K1 cells, pancreatic Min-6 cells, and cardiomyocyte H9c2 cells. The results indicated that the antihyperglycemic effect of SA in diabetic rats was abolished by blocking the GLP-1 receptor with an antagonist. SA has a direct effect on the GLP-1 receptor when using CHO-K1 cells transfected with the exogenous GLP-1 receptor gene. In addition, SA stimulated insulin production in Min-6 cells by activating GLP-1 receptors. SA caused a dose-dependent rise in GLP-1 receptor mRNA levels in cardiac H9c2 cells. These in vitro results support the notion that SA has a direct effect on the GLP-1 receptor. Otherwise, SA inhibited the increase of pro-inflammatory cytokines, including interleukins and tumor TNF-α, in type 1 diabetic rats in a dose-dependent manner. Moreover, as with liraglutide, SA reduced plasma lipid profiles, including total cholesterol and triglyceride, in mixed diet-induced type 2 diabetic rats. Intriguingly, chronic treatment with SA (as with liraglutide) reversed the functions of both the liver and the kidney in these diabetic rats. SA displayed less efficiency in reducing body weight and food consumption compared to liraglutide. In conclusion, SA effectively activates GLP-1 receptors, resulting in a reduction in diabetic-related complications in rats. Therefore, it is beneficial to develop SA as a chemical agonist for clinical applications in the future.

Oil droplet fouling on lesser sandeel (Ammodytes marinus) eggshells does not enhance the crude oil induced developmental toxicity

The oil industry's expansion and increased operational activity at older installations, along with their demolition, contribute to rising cumulative pollution and a heightened risk of accidental oil spills. The lesser sandeel (Ammodytes marinus) is a keystone prey species in the North Sea and coastal systems. Their eggs adhere to the seabed substrate making them particularly vulnerable to oil exposure during embryonic development. We evaluated the sensitivity of sandeel embryos to crude oil in a laboratory by exposing them to dispersed oil at concentrations of 0, 15, 50, and 150 µg/L oil between 2 and 16 days post-fertilization. We assessed water and tissue concentrations of THC and tPAH, cyp1a expression, lipid distribution in the eyes, head and trunk, and morphological and functional deformities. Oil droplets accumulated on the eggshell in all oil treatment groups, to which the embryo responded by a dose-dependent rise in cyp1a expression. The oil exposure led to only minor sublethal deformities in the upper jaw and otic vesicle. The findings suggest that lesser sandeel embryos are resilient to crude oil exposure. The lowest observed effect level documented in this study was 36 µg THC/L and 3 µg tPAH/L. The inclusion of these species-specific data in risk assessment models will enhance the precision of risk evaluations for the North Atlantic ecosystems.

Dickkopf-related Protein 2 Promotes Hair Growth by Upregulating the Wnt/β-catenin Signaling Pathway in Human Dermal Papilla Cells.

The Wnt/β-catenin signaling pathway is crucial for the development, initiation, and growth of hair follicles (HFs). The Dickkopf-related protein (DKK) gene family encodes secreted proteins modulating the Wnt/β-catenin signaling pathway. Studies have reported that DKK1 promotes the regression of HFs and serves as a pathogenic mediator in male pattern baldness. However, the role of DKK2 on human hair growth has not yet been explored. This study investigates direct effect of DKK2 on hair growth using human dermal papilla cell (DPC) cultures and ex vivo human HF organ cultures. To elucidate the effect of DKK2 on hair growth, we examined the effect of recombinant human DKK2 (rhDKK2) treatment on cell viability, expression of mRNA and protein related to the Wnt/β-catenin signaling pathway, and cell growth in cultured human DPCs. We also performed ex vivo organ culture of HFs with rhDKK2 and measured changes in hair shaft length for 8 days. Treatment with rhDKK2 led to a dose-dependent rise in the proliferation of human DPCs (p<0.05), reaching levels comparable to those induced by 1 μM minoxidil. Moreover, rhDKK2 increased the expression of Wnt/β-catenin target genes, phosphorylated extracellular signal-regulated kinase, and cyclin-D1; it also increased the BAX-to-Bcl-2 ratio and downregulated the bone morphogenetic protein 2 gene. In human HF organ cultures, relative to the control treatment, rhDKK2 treatment significantly increased hair shaft elongation (p<0.01). Our results indicate that rhDKK2 could promote hair growth by facilitating the proliferation of human DPCs through activation of the Wnt/β-catenin signaling pathway.

In-vivo investigation of dimethoate toxicity on serum enzymes, target organs and intestinal tissues of albino rats

Dimethoate is a broad spectrum organophosphate insecticide used to control agricultural, industrial and domestic pests. Despite its benefits, toxicities in non-target organisms have been widely reported. This study investigated acute and sub-chronic toxicities of dimethoate in rats. The arithmetic method of Karbar and a modification of repeated 8-week oral toxicity were adopted for the determination of acute and sub-chronic toxicities of dimethoate, respectively. Symptoms of acute toxicity evaluated include muscular weakness, respiratory distress, convulsion and death. Animals that were used for the sub-chronic toxicity studies were divided into 4 groups of 5. Groups 2, 3 and 4 were fed with feeds containing 200, 500 and 800 mg/kg dimethoate respectively, while group 1 served as the control. The weight of the animals were determined weekly for 8 weeks, and thereafter sacrificed. Blood samples were collected for biochemical tests, and the liver, kidney, lung, intestine and heart were excised and processed for histopathological analysis. The calculated median lethal dose (LD50) was 176 mg/kg. Animals in all groups gained weight progressively. However, a dose-dependent significant rise in alanine transaminase and alkaline phosphatise, but not aspartate transaminase accompanied with significant focal necrosis in liver, eosinphilic casts in kidney and intestinal ulceration in group 3 and 4 in animals. In addition, dimethoate- induced inflammation was observed in the liver, kidney, lung and intestine tissues. The derangement of biochemical parameters and relevant histopathological alterations observed in rats exposed to dimethoate are suggestive of toxicity. Therefore necessary precautionary measures should be taking during handling and use.

Long-Term Persistence of Antibodies to Adeno-Associated Viral Vectors Following Gene Therapy with scAAV8-LP1-Fixco

Background Gene therapy for hemophilia B (HB) consisting of a one-time, systemic, administration of adeno-associated viral (AAV) vectors encoding a functional Factor-IX gene is now approved and represents an important and long-awaited milestone. A rise in anti-AAV antibody titers follows systemic administration of AAV vectors. This does not have direct clinical consequences; however, persistence of neutralising anti-AAV antibodies (NAB) at high titers will preclude subsequent successful gene transfer with vector of the same serotype. This would be problematic, in the event that transgene expression should fall below therapeutic levels and repeat dosing might be needed. Furthermore, high titer NAB response to one serotype can cross-react with other serotypes thereby limiting the option of using alternative serotype AAV vectors. We report a longitudinal analysis of anti-AAV antibody response over a 10-year period in severe HB participants following systemic administration of scAAV2/8-LP1-hFIXco from the St Jude-UCL Phase I/II, gene therapy trial (AGT4HB ClinicalTrials.gov:NCT00979238). Methods Anti-AAV antibody levels were evaluated in the 10 trial participants following a bolus intravenous infusion of scAAV2/8-LP1-hFIXco at a dose of 2x10 11vg/kg (N=2), 6x10 11vg/kg (N=2), or 2x10 12vg/kg (N=6) between 2010 and 2012. Total anti-AAV antibodies (TAB) were measured by ELISA, and levels quantitated using a pooled IgG reference. NAB were quantitated by an in-vitro transduction inhibition assay (TIA) in which plasma samples were preincubated with AAV CMV NanoLuc (MOI: AAV3 1, AAV5 100 and 8 100) prior to evaluation of transduction of HEK 293T cells. Results were normalised to virus control and the 50% inhibition dilution (IC50) was calculated. For comparison, we assessed anti-AAV antibody titers in 38 normal human plasma samples using the same methodology. Results At the time of data cut-off (31 st Dec 2022) the median follow-up for the cohort was 10.7 years (range: 4-12 years) with follow-up in two participants (one mid dose and the other high dose) being limited to 4 years. A dose dependent AAV8 TAB response was observed with low dose participants having a mean of 84±101mg/ml at 1-year, mid dose 507 mg/ml and high dose 1650±603 mg/ml. This represented a &amp;gt;1400-fold increase from baseline, pre-AAV administration values. These TAB levels were significantly higher than levels following natural infection in 38 seropositive individuals not exposed to recombinant vector (normal range 0.31-34.02mg/ml). In the high dose cohort, TAB titer against AAV3 and AAV5 at 1 year after gene transfer were 145±167 mg/ml (natural infection reference range 0.31-29.61mg/ml) and 96±55 mg/ml (normal range 0.31-36.13mg/ml) respectively, consistent with cross-reactivity between AAV serotypes. There was a decline in TAB levels against all serotypes prior to reaching a plateau. At year 5 the high dose cohort was 928.9 ±556.9mg/ml for AAV8, 90.4±95.6 mg/ml for AAV3 and, 72.8 ±49.1 mg/ml for AAV5. AAV8 NAB titers had a similar kinetic with IC50 at 1 year 7261.5 ±9812 (low-dose), 15000 (mid-dose) and, 40204± 37823 (high dose). For comparison, AAV8 IC50 were significantly higher than levels following natural infection with wild-type AAV in seropositive individuals (range 5-747) (Mann Whitney p&amp;lt;0.0001). NABs levels declined to an IC50 level of 12907±8246 at 5 years for the high dose cohort, which was still 3000-fold higher than their baseline value (range 5-70.1) (Mann Whitney p&amp;lt;0.0001), and above the threshold for efficient gene transfer. The development of cross-reactive NAB levels was relatively modest and within the range observed in natural seropositive individuals. For the high dose cohort NAB to AAV3 at year 1 was 6536± 1869 (normal range 5- 13502), and for AAV5 at year 1 852± 596 (normal range 5-2143). Conclusion Following AAV gene therapy there is a robust dose dependent rise in anti-AAV 8 antibody levels that decline over time, but at 10 years are still significantly above the threshold for re-administration of vector of the same serotype. There was clear evidence of cross reactivity against AAV3 and 5. AAV5 NAB titres were lower than other serotypes declining to levels below a threshold required for efficient gene transfer in the low and mid dose cohort within 10 years. This suggests that, should FIX levels fall below therapeutic levels in this cohort, successful re-administration of an alternative vector may be possible.

Surface-Modified Silver Nanoparticles and Their Encapsulation in Liposomes Can Treat MCF-7 Breast Cancer Cells.

Silver nanoparticles (AgNPs) have emerged as a promising tool for cancer treatment due to their unique physicochemical and biological properties. However, their clinical applications are limited by their potential cytotoxicity caused due to oxidation stress and non-specific cellular uptake pathways. To overcome these barriers, surface modifications of AgNPs have been proposed as an effective strategy to enhance their biocompatibility and specificity toward cancer cells. In this study, AgNPs were synthesised using the chemical reduction method and subsequently conjugated with various capping agents such as Polyvinylpyrrolidone (PVP) and Bovine Serum Albumin (BSA). Further, this study involves the synthesis of liposomes by using dipalmitoyl phosphatidylcholine lipid (DPPC) and cholesterol to increase the biocompatibility and bioavailability of AgNPs to MCF-7 breast cancer cells. In vitro, cytotoxicity studies were performed to determine which surface modification method exhibited the highest cytotoxic effect on the MCF-7 breast cancer cells, which was determined through the MTT assay. The AgNPs conjugated with BSA exhibited the highest cytotoxicity at the lowest dosage, with an IC50 of 2.5 μL/mL. The BSA-AgNPs induced a dose-dependent rise in cytotoxicity through the enhancement of nucleophilic dissolution of the AgNPs in cancer cells. In comparison, the unmodified AgNPs had an IC50 value of 3.0 μL/mL, while the PVP-modified AgNPs had an IC50 of 4.24 μL/mL. AgNPs encapsulated in liposomes had an IC50 value of 5.08 μL/mL, which shows that the encapsulation of AgNPs in liposomes controls their entry into cancer cells. The findings of this research have provided insights into the potential use of surface-modified AgNPs and liposomal encapsulated AgNPs as novel therapeutic tools to overcome the conventional treatment limitations of breast cancer cells.

Assessment of the hypothalamic-pituitary-adrenocortical axis function using a vasopressin stimulation test in neonatal foals.

Bacterial sepsis is the leading cause of death in foals and is associated with hypothalamic-pituitary-adrenocortical axis (HPAA) dysfunction. HPAA function can be evaluated by an arginine-vasopressin (AVP) stimulation test. Administration of AVP will stimulate a dose-dependent rise in systemic adrenocorticotropin-releasing hormone (ACTH) and cortisol in neonatal foals. There will be no response seen in corticotropin-releasing hormone (CRH) and baseline AVP will be within reference interval. Twelve neonatal foals, <72 hours old. HPAA function was assessed in foals utilizing 3 doses of AVP (2.5, 5, and 7.5 IU), administered between 24 and 48 hours of age in this randomized cross-over study. Cortisol, ACTH, CRH and AVP were measured at 0 (baseline), 15, 30, 60 and 90 minutes after AVP administration with immunoassays. The fold increase in cortisol and ACTH was calculated at 15 and 30 minutes compared to baseline. All doses of AVP resulted in a significant increase in cortisol concentration over time, and a dose-dependent increase in ACTH concentration over time. ACTH and cortisol were significantly increased at 15 and 30 minutes, respectively after all 3 doses of AVP compared to baseline (P < .01). There was no change in endogenous CRH after stimulation with AVP. Administration of AVP is safe and results in a significant rise in ACTH and cortisol in neonatal foals. A stimulation test with AVP (5 IU) can be considered for HPAA assessment in septic foals.

Composition and Biological Activity of Fetoplacental Tissues-Derived Cryoextracts Being Differently Obtained

The protein-peptide composition of fetal tissues (FTCEs) and placenta (PCE) cryoextracts of rats was investigated by gel permeation chromatography. Cryoextracts were derived from tissue homogenates using the freeze-warming modes: 1 – single (–20°С); 2 – double (–20; –196°С) and 3 – triple (–20; –196; –196°С) ones. The biological activity of cryoextracts was in vitro evaluated by the phagocytic activity of neutrophil granulocytes (NG) of the blood of intact rats after incubation with an inactivated culture of Staphylococcus aureus (2 × 109 cells/ml) for 45 and 120 min for the concentrations of extracts in the incubation medium: 1.6; 3.1; 6 mg/ml in terms of dry substance. The use of mode 3 ensured a greater yield of total protein and low molecular weight compounds of a protein-peptide nature in the composition of cryoextracts. The content of low-molecular-weight fractions in FTCE exceeded that in PCE, regardless of the obtaining mode. Incubation of NG with cryoextracts did not significantly increase the number of neutrophils which entered into phagocytosis. However, all the samples of PCE and FTCE were characterized by a dose-dependent rise in the NG absorption activity during the 45-minute incubation compared to the control.

Elucidation of the role of α-lipoic acid and vitamin C in methotrexate-induced hepatoxicity in mice.

Although methotrexate (MTX) is used to treat several malignancies and chronic inflammatory diseases, its clinical use is constrained because of its negative side effects, the most prevalent of which are hepatotoxicity and nephrotoxicity. So, this study aims to determine whether α-lipoic acid (ALA) and vitamin C can protect mice against the liver damage that methotrexate causes. A total of 49 male mice were divided into seven groups at random. Group I received sodium bicarbonate, while groups II to VII received an intraperitoneal injection of MTX (20 mg/kg) on the tenth day, following ten days of pretreatment with ALA (60 mg/Kg), ALA (120 mg/Kg), vitamin C (100 mg/Kg), vitamin C (200 mg/Kg), ALA (60 mg/Kg), and vitamin C (100 mg/kg). When compared to mice in group I, mice in group II (the control group) had significantly higher levels of the enzymes malondialdehyde (MDA), alanine transaminase (ALT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) and significantly lower (p <0.05) levels of the enzymes superoxide dismutase (SOD) and glutathione (GSH). As compared to the control group, pretreatment groups with ALA and vitamin C showed a dose-dependent substantial rise (p <0.05) in GSH and SOD levels, a dose-dependent notable decrease (p <0.05) in MDA, ALT, ALP, and LDH levels, and better liver histological architecture. In order to increase the antioxidant capacity, pretreatment with ALA and vitamin C may be able to prevent MTX-induced hepatotoxicity. These results imply that ALA and vitamin C are useful in the treatment of MTX-induced liver damage.

Quantification of Xanthone and Anthocyanin in Mangosteen Peel by UPLC-MS/MS and Preparation of Nanoemulsions for Studying Their Inhibition Effects on Liver Cancer Cells

Mangosteen peel, a waste produced during mangosteen processing, has been reported to be rich in xanthone and anthocyanin, both of which possess vital biological activities such as anti-cancer properties. The objectives of this study were to analyze various xanthones and anthocyanins in mangosteen peel by UPLC-MS/MS for the subsequent preparation of both xanthone and anthocyanin nanoemulsions to study their inhibition effects on liver cancer cells HepG2. Results showed that methanol was the optimal solvent for the extraction of xanthones and anthocyanins, with a total amount of 68,543.39 and 2909.57 μg/g, respectively. A total of seven xanthones, including garcinone C (513.06 μg/g), garcinone D (469.82 μg/g), γ-mangostin (11,100.72 μg/g), 8-desoxygartanin (1490.61 μg/g), gartanin (2398.96 μg/g), α-mangostin (51,062.21 μg/g) and β-mangostin (1508.01 μg/g), as well as two anthocyanins including cyanidin-3-sophoroside (2889.95 μg/g) and cyanidin-3-glucoside (19.72 μg/g), were present in mangosteen peel. The xanthone nanoemulsion was prepared by mixing an appropriate portion of soybean oil, CITREM, Tween 80 and deionized water, while the anthocyanin nanoemulsion composed of soybean oil, ethanol, PEG400, lecithin, Tween 80, glycerol and deionized water was prepared as well. The mean particle size of the xanthone extract and nanoemulsion were, respectively, 22.1 and 14.0 nm as determined by DLS, while the zeta potential was -87.7 and -61.5 mV. Comparatively, xanthone nanoemulsion was more effective than xanthone extract in inhibiting the growth of HepG2 cells, with the IC50 being 5.78 μg/mL for the former and 6.23 μg/mL for the latter. However, the anthocyanin nanoemulsion failed to inhibit growth of HepG2 cells. Cell cycle analysis revealed that the proportion of the sub-G1 phase followed a dose-dependent increase, while that of the G0/G1 phase showed a dose-dependent decline for both xanthone extracts and nanoemulsions, with the cell cycle being possibly arrested at the S phase. The proportion of late apoptosis cells also followed a dose-dependent rise for both xanthone extracts and nanoemulsions, with the latter resulting in a much higher proportion at the same dose. Similarly, the activities of caspase-3, caspase-8 and caspase-9 followed a dose-dependent increase for both xanthone extracts and nanoemulsions, with the latter exhibiting a higher activity at the same dose. Collectively, xanthone nanoemulsion was more effective than xanthone extract in inhibiting the growth of HepG2 cells. Further research is needed to study the anti-tumor effect in vivo.

CENTELLA ASIATICA NEUROPROTECTIVE EFFECT ON 6-OHDA-STIMULATED OXIDATIVE STRESS IN DIFFERENTIATED SH-SY5Y CELLS

Aim: Parkinson's disease is qualified by advancing the loss of dopaminergic neurons and depletion of dopamine. However, the pathophysiological mechanisms need new perspectives for therapeutic strategies that alleviate and abolish neurodegenerative. The last searches have demonstrated that Centella Asiatica is commonly used in conventional medicine. The aim of our study is to reveal the neuroprotective effect of Centella Asiatica, which we will use in the treatment, on the neurotoxicity stimulated by 6 OHDA. Method: First, the SH-SY5Y cell line was grown in prepared media. Then, 25-50-75 and 100 µg/ml concentrations of Centella Asiatica were supplemented to the wells that reached 85% confluence 2 hours before (except for the control and 6-OHDA groups). Afterward, 200 µM 6-OHDA was added to the wells (except for the control group) and incubated for 24 hours. Then, IL-1β, GSH, MTT, LDH, GPx, TNF-α, SOD, MPO, CAT, and MDA analyses were performed. One-way analysis of variance was performed using the IBM SPSS 22.0 package program. The results were compared with the control and 6-OHDA groups, and values below p&lt;.05 were considered statistically significant. Results: It was found that Centella Asiatica demonstrated a dose-dependent rise in the vivacity rate and the cell vivacity was 92% at the highest concentration. Moreover, antioxidant parameters (GSH, GPx, SOD, CAT) correlated with MTT and LDH assay. In IL-1β, TNF-α, MPO, and MDA activities, it was observed that the oxidant amount reduced depending on the concentration. Conclusion: These findings revealed that Centella Asiatica exerts a neuroprotective effect opposite 6-OHDA induction by rising cell viability and reducing oxidative stress.

Investigating the anticancer efficacy of biogenic synthesized MgONPs: An in vitro analysis.

The biogenic approach of synthesizing metal nanoparticles is an exciting and interesting research area with a wide range of applications. The present study reports a simple, convenient, low-cost method for synthesizing magnesium oxide nanoparticles (MgONPs) from pumpkin seed extracts and their anticancer efficacy against ovarian teratocarcinoma cell line (PA-1). The characteristic features of biogenic MgONPs were assessed by UV–visible spectrophotometry (UV–vis), X-ray powder diffraction (XRD), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The formation of spherical NPs with an average size of 100 nm was observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Moreover, MgONPs exhibit considerable cytotoxicity with an IC50 dose of 12.5 μg/ml. A dose-dependent rise in the induction of apoptosis, ROS formation, and inhibition in the migration of PA-1 cells was observed up to 15 μg/ml concentration, reflecting their significant anticancer potential against ovarian teratocarcinoma cell line. However, additional work, especially in different in vitro and in vivo models, is recommended to find out their real potential before this environment-friendly and cost-effective nanoformulation could be exploited for the benefit of humankind.

In Vitro Anti-Inflammatory Evaluation of Sesbania Procumbens Extracts

Plants used in the traditional healing system require a systematic evaluation that may give more promising data about their medicinal value and may be useful to meet the rising demand for novel agents to combat infections and diseases. The plant Sesbania procumbens is a one used traditionally by the rural population for a varietyof medicinal purposes including anti inflammatory activity. Based on this report the plant S. procumbens was selected for our research and in this study different extracts of S. procumbens were evaluated in vitro for anti-inflammatory activity. The whole plant material was collected, authenticated, aerial parts were separated, shade dried for about two weeks and made into a coarse powder by using mechanical grinder. Extraction of powdered material was done by soxhlation using the different solvents of increasing polarity viz., petroleum ether, chloroform, ethyl acetate and methanol. Dried extracts thus obtained were subjected to in vitro anti-inflammatory evaluation by human red blood cell (HRBC) membrane stabilization in normal and different temperature (54°C and −10°C) and tonicity (hypotonic and isotonic) conditions. Six different concentrations (100, 200, 400, 600, 800, 1000μg/ml) of four extracts were subjected to evaluation with indomethacin (100μg/ml) as standard control. The data thus collected was analysed by using SPSS version 15.0. All values are expressed as mean ± SD. 0.05 was considered statistically significant From the results, a dose-dependent rise in antiinflammatory activity was found in all the tested extracts. Particularly, the methanol and ethyl acetate extracts showed significant activity in their maximum testing concentration (1000µg/ml) in all the experimental approaches. The results of this study strongly supported the claims of the anti-inflammatory property of the tested extracts. Further studies such as in vivo evaluations and phytochemical screening of the extracts may give more significant data which may be useful for the development of novel chemotherapeutic agents

Distribution of Fluoride in Plasma, Brain, and Bones and Associated Oxidative Damage After Induced Chronic Fluorosis in Wistar Rats.

The study was aimed to determine fluoride levels in plasma, brain, and bones of Wistar rats following chronic administration of fluoride at different dose levels and the consequent oxidative damage inflicted in these tissues. Brain histomorphology and bone radiographs were also evaluated to assess the extent of damage in these organs. Eighteen rats were randomly divided into three groups with six animals in each group. Group I served as control and groups II and III received 50 and 100ppm fluoride in tap water, respectively for 180days. A dose-dependent rise in the levels of fluoride in plasma, brain, and bones was observed in rats. Significant (P < 0.05) alterations in levels of total thiols, glutathione peroxidase, glutathione reductase, acetylcholinesterase, catalase, superoxide dismutase, lipids, as well as protein peroxidation in blood and brain were observed as compared to control in a dose-dependent manner. Radiological examination of bone revealed thinning of bone cortex with haphazard ossification, reduced bone density, and widening of marrow cavity indicating occurrence offlawed bone remodeling upon chronic fluoride exposure. Improper mineralization in bones of intoxicated rats indirectly reflected reduced bone tensile strength. Moreover, alterations in plasma Ca:P ratio and high levels of fluoride in bone ash indicated that chronic fluoride exposure leads to alterations in the bone matrix further corroborating the radio-graphical findings. Additionally, severe microscopic alterations were recorded in the cerebrum and cerebellum of treated rats which included neuronal necrosis, gliosis, spongiosis, perivascular cuffing, congestion, and hemorrhage which correlated well with oxidative changes induced by fluoride intoxication in the brain tissue of rats.

Alpelisib-Induced Hyperglycemia- A Small Case Series

The phosphoinositide-3-kinase (PI3K) family consists of highly conserved enzymes that are key intermediaries in signal transduction regulating cell survival and mitogenesis. This axis has been implicated in many types of cancers. In 2019 Alpelisib (Piqray), a reversible inhibitor specific to the PI3Kα subunit, in combination with fulvestrant, was approved to treat hormone receptor positive, human epidermal growth factor receptor 2 (Her2) negative, PI3K mutated breast cancer. This approval followed SOLAR-1 trial results showing a 35% risk reduction in cancer progression or death with alpelisib-fulvestrant compared to placebo in a cohort with PI3K mutated breast cancer. PI3K also plays a critical role in the insulin signaling pathway and alpelisib has been shown to cause a dose dependent rise in plasma glucose, insulin and c-peptide. Here we present a case series of severe hyperglycemia induced by alpelisib.A 44-year-old woman with ER+/Her2(-)/PI3K positive metastatic breast cancer was started on alpelisib. Previously, HbA1C was 5.4%. Hyperglycemia developed and HbA1c rose to 9.0% within 6 months of alpelisib 300mg daily. She started metformin and empagliflozin, which she was unable to tolerate due to nausea and vomiting. Her self-monitored blood glucoses were 300-400mg/dL within hours after her morning alpelisib dose. We discontinued empagliflozin when she developed metabolic acidosis with an increased anion gap. However, prior to any dose reduction, oncology discontinued alpelisib due to evidence of cancer progression. A week later, her glucoses normalized.Second case is of a 64-year-old woman with stage IV ER+/Her2(-)/PI3K mutated breast cancer with bony metastases, who was started on alpelisib 250mg. Her prior HbA1C was 5.5%. Ten days after initiation of alpelisib, she developed grade 3 hyperglycemia (blood glucoses 200-500mg/dL). She was started on metformin 2000mg with alpelisib dosed at noon. However, she noted a marked rise in blood glucose in the afternoon, few hours following alpelisib dose. Thus, moving the alpelisib to bedtime allowed better control of glycemia by using overnight basal insulin.Similarly, a 37-year-old woman with a history of ER+/HER2(-) stage IV metastatic breast cancer to the liver, with PI3K mutation was found to have acute, severe hyperglycemia with blood glucose of 300mg/dL, despite HbA1C being only 4.7%. This was attributed to initiation of alpelisib 2 days prior to admission. Given the severity of her insulin resistance (requiring > 100 units of insulin daily), alpelisib dose was reduced from 300mg to 150mg/day. On discharge, she was placed on metformin, dulaglutide, and basal and prandial insulins. Her HbA1C rose to 9.4% within 3 months of alpelisib initiation.This case series demonstrate the unique challenges in managing alpelisib induced reversible hyperglycemia.

Effect of Engineered Nickel Oxide Nanoparticle on Reactive Oxygen Species-Nitric Oxide Interplay in the Roots of Allium cepa L.

Scientists anxiously follow instances of heavy metals augmenting in the environment and undergoing bioaccumulation and trace their biomagnification across food webs, wary of their potent toxicity on biological entities. Engineered nanoparticles supplement natural pools of respective heavy metals and can mimic their effects, exerting toxicity at higher concentrations. Thus, a thorough understanding of the underlying mechanism of this precarious interaction is mandatory. Most urban and industrial environments contain considerable quantities of nickel oxide nanoparticles. These in excess can cause considerable damage to plant metabolism through a significant increase in cellular reactive oxygen species and perturbation of its cross-talk with the reactive nitrogen species. In the present work, the authors have demonstrated how the intrusion of nickel oxide nanoparticles (NiO-NP) affected the exposed roots of Allium cepa: starting with disruption of cell membranes, before being interiorized within cell organelles, effectively disrupting cellular homeostasis and survival. A major shift in the reactive oxygen species (ROS) and nitric oxide (NO) equanimity was also observed, unleashing major altercations in several crucial biochemical profiles. Altered antioxidant contents and upregulation of stress-responsive genes, namely, Catalase, Ascorbate peroxidase, Superoxide dismutase, and Rubisco activase, showing on average 50–250% rise across NiO-NP concentrations tested, also entailed increased cellular hydrogen peroxide contents, with tandem rise in cellular NO. Increased NO content was evinced from altered concentrations of nitric oxide synthase and nitrate reductase, along with NADPH oxidase, when compared with the negative control. Though initially showing a dose-dependent concomitant rise, a significant decrease of NO was observed at higher concentrations of NiO-NP, while cellular ROS continued to increase. Modified K/Na ratios, with increased proline concentrations and GABA contents, all hallmarks of cellular stress, correlated with ROS–NO perturbations. Detailed studies showed that NiO-NP concentration had a significant role in inducing toxicity, perturbing the fine balance of ROS–NO, which turned lethal for the cell at higher dosages of the ENP precipitating in the accumulation of stress markers and an inevitable shutdown of cellular mechanisms.