Significant Dose-dependent Increase Research Articles

The Dose-Dependent Effect of Obesity on Adverse Maternal and Neonatal Outcomes in a Hispanic Population.

To evaluate the frequency of adverse maternal and neonatal outcomes associated with maternal obesity in a Hispanic population. We hypothesized that obesity confers a dose-dependent risk associated with these outcomes. This was a retrospective cohort study of singleton pregnancies delivered between 24 and 42 weeks gestation at an urban county hospital between 2013 and 2021. Body mass index (BMI) at first prenatal visit was used as a proxy for pre-pregnancy weight. Patients were excluded if their first trimester BMI was not available. Trends in adverse outcomes across increasing obesity classes were assessed. During the study period, 58,497 patients delivered a singleton infant, of which 12,365 (21.1%), 5429 (9.3%), and 3482 (6.0%) were in class I, II, and III obesity, respectively. Compared with non-obese patients, more obese patients were more likely to be younger and nulliparous with a higher incidence of hypertension and pregestational diabetes. Higher BMI was associated with a significant dose-dependent increase in cesarean delivery (27% for non-obese, 34% for class I, 39% for class II, and 46% for class III obesity); severe preeclampsia (8% in non-obese and 19% for class III obesity); and gestational diabetes (5% in non-obese and 15% in class III obesity). There were significant trends in increasing morbidity for infants born to patients with correspondingly higher obesity classes. Some of these adverse outcomes included respiratory distress syndrome, neonatal intensive care unit admission, fetal anomalies, and sepsis (all p < 0.001). Increasing body mass index is associated with a significant dose-dependent increase in multiple adverse perinatal outcomes in a Hispanic population. Associated adverse maternal outcomes include severe preeclampsia, gestational diabetes, and cesarean delivery. Infants born to patients with correspondingly higher BMI class have significantly increased associated morbidity. Often, only higher BMI classes are significantly associated with these adverse outcomes.

In Vivo Evaluation of Genotoxicity and Antioxidant Capacity of Ajuga Orientalis L. (Lamiaceae) Leaf Extracts

Ajuga orientalis L. (Lamiaceae) is a fragrant herb native to the Eastern Mediterranean region, widely used in traditional healing practices in Jordan and neighboring countries. Despite its extensive use, there is a lack of toxicological studies on its leaf extracts. This study aims to address this gap by evaluating the genotoxic potential of ethanolic and aqueous leaf extracts using a micronucleus (MN) assay on mice DNA, alongside assessing their antioxidant status. The median lethal dose 50% (LD50) was tested in ten groups of sixty male Balb/c mice to determine the acute toxicity of A. orientalis leaf extracts. Four groups of male Balb/c mice (n=6) were used to evaluate micronucleus (MN) formation and total antioxidant capacity for each extract. Each group received daily intraperitoneal injections of one of the following concentrations: 4000, 2000, 1000, and 500 mg/kg over 28 days. Additionally, three control groups were included for comparison purposes. Peripheral blood samples were screened for MN formation, and liver samples were assessed for total antioxidant capacity. Results revealed an LD50 of 4000 mg/kg for both extracts, alongside a significant dose-dependent increase in MN formation and lower antioxidant capacity compared to controls. The findings indicate the genotoxicity of A. orientalis leaf extracts in Balb/c mice, urging caution in human consumption. Further research is warranted to comprehensively assess their safety and toxicity, especially considering their traditional medicinal use.

Targeting the SIRT3/MnSOD and JNK/HMGB1/Beclin 1 Axes: Role of Apigenin in Multifaceted Metabolic Intervention in Colorectal Cancer.

Colorectal cancer (CRC) is the third most prevalent cancer worldwide. While chemotherapy remains the standard treatment approach, natural products have emerged as a promising alternative. Among these, apigenin, a natural flavonoid, has garnered significant attention due to its pro-oxidant and antioxidant properties in various types of cancer. This study aimed to assess the potential impact of apigenin in CRC treatment by targeting mitochondrial SIRT3, HMGB1, and beclin 1-mediated autophagy in a mouse model of CRC. We administered 20 mg/kg of dimethyl hydrazine (DMH) intraperitoneally once weekly for 20 weeks to induce CRC in C57BL/6 mice. After 6 weeks of initiating the study, apigenin was intragastrically co-administered by oral gavage at 25 and 50 mg/kg until the end of week 20. The results revealed significant weight loss, shortening of the colon, and diarrhea in DMH-induced CRC, which are considered the marks of CRC. In addition, histopathological examination revealed dysplastic changes in the DMH-treated group, while no dysplasia was found in the apigenin-treated CRC groups. Importantly, the administration of apigenin to DMH-treated animals has led to a significant reduction of SIRT3 and MnSOD expression levels with a significant increase in LC3-II at either dose and a significant dose-dependent increase in the levels of MDA, c-JNK, HMGB1, and beclin 1 compared to the DMH-treated group. In conclusion, apigenin may have a promising role in suppressing DMH-induced CRC. It elicits a pro-oxidant activity by suppressing the gene expression of SIRT3 and subsequently, its target MnSOD, resulting in increased reactive oxygen species (ROS) and lipid peroxidation. The released ROS, in turn, activates JNK-mediated autophagy by enhancing HMGB1, beclin 1, and LC3-II protein levels.

Evaluation of the toxic potential of ethyl methanesulphonate (EMS) on Hydra vulgaris

The effect of EMS at final concentration of 0.09, 0.18, 0.27 and 0.37mM was studied on Hydra vulgaris using morphological, regeneration, oxidative stress markers and DNA damage as parameters. The morphological scores showed a significant dose dependent difference in the Hydra exposed to 0.18, 0.27, and 0.37mM of EMS for 24, 48, 72 and 96 hrs. The regeneration scores also showed a significant difference in the gastric region of Hydra exposed to 0.37mM of EMS for 48hr. A significant difference in the scores of regeneration was observed for the mid body portion exposed to 0.18, 0.27 and 0.37mM of EMS for 72 and 96hr of duration compared to control. A dose-dependent significant increase in the activities of glutathione-S-transferase (GST), catalase (CAT), and superoxide dismutase (SOD) was observed compared to control. The thiobarbituric acid reactive species (TBARS) levels were also significantly increased compared to control. The genotoxic damage was assessed in the cells of gastric region of the Hydra exposed to 0.09, 0.18, 0.27 and 0.37mM of EMS for 48hr by performing comet assay. A significant dose-dependent increase in the DNA damage was observed compared to control.

EFFECTS OF THE AQUEOUS BARK EXTRACT FROM STRYCHNOS CAMPTONEURA AGAINST 40% ETHANOL-INDUCED MALE INFERTILITY IN MALE GUINEA PIGS(CAVIAPORCELLUS)

The overall objective of this study was to assess the effects of the aqueous extract derived from the bark of Strychnos camptoneura on reproductive functions in male guinea pigs (Cavia porcellus) following ethanol 40% intoxication. To assess these effects, 20 guinea pigs were randomly sorted into 4 lots, each containing 5 animals and given distilled water at a dose of 1ml/kg for lot1 ethanol at a dose of 1ml/kg for lot 2 (positive controls) and ethanol at a dose of 1ml/kg for lot 3: ethanol at 1ml/kg and aqueous extract fromStrychnos camptoneura trunk bark at 100mg/kg and for lot 4: ethanol at 1ml/kg and aqueous extract from Strychnos camptoneura bark, given at a dosage of250mg/kg. The results demonstratedthat oral administration of aqueous extract fromStrychnos camptoneura trunk bark (100mg/kg and 250mg/kg) in male guinea pigs (Cavia porcellus) after ethanol intoxication 40% moderately stimulates food consumption compared with positive controls given ethanol exclusively. Results demonstrated that oral administration of aqueous extract fromStrychnos camptoneura trunk bark (100 and 250 mg/kg) in ethanol-treated male guinea pigs causes a non-significant (P ˃ 0.05) increase in the weight of vas deferens, seminal vesicles and penis, compared with negative controls given distilled water and positive controls treated exclusively with ethanol.In addition, gavage of the animals at the doses studied produced a significant (P&lt;0.01) dose-dependent increase in testicular and epididymal weights, in contrast to the positive controls, which showed a significant (P&lt;0.01) decrease in these two sexual organs. Treatment with the extract did not result in significant changes (P˃0.05)in the weight of the vas deferens, seminal vesicles, or penis. Oral administration of the aqueous extract of Strychnos camptoneura trunk bark (100 and 250mg/kg) to ethanol-exposed male guinea pigs resulted in highly and highly significant (p&lt;0.01 p&lt;0.001) dose-dependent increases in sperm concentration per epididymal tail compared with lots treated with distilled water and ethanol alone. In addition, there was a highly significant (p&lt; 0.001) dose-relatedincrease in the rate of motile and vital spermatozoa, compared with negative and positive controls given distilled water and ethanol alone.In contrast, a highly significant (p&lt; 0.01 p&lt; 0.001) decrease in sperm count, motility rate and vitality was observed in male guinea pigs treated with ethanol alone. These results show that gavage of the animals with the extract, at the doses studied after exposure to ethanol, results in a non-significant (p˃ 0.05) increase in serum testosterone levels, compared with positive controls. In addition, positive ethanol-only controls were found to experience a significant (p&lt;0.01) decrease in testosterone levels when compared to negative controls receiving distilled water. These results provide evidence that the extract may offer protective benefitsgainst alcohol-induced male infertility, confirming its androgenic and spermatogenic potential.

Neuroprotective Role of Ranolazine: ESR1 and NMDA Receptor Agonist in Traumatic Brain Injury in Drosophila melanogaster, In Silico and In Vivo Correlation

Abstract Objective In this study, a high-impact trauma (HIT) device was used for inducing moderate traumatic brain injury (TBI) in Drosophila melanogaster. Mechanical injuries in flies caused by rapid acceleration and assertion produce symptoms characteristics of TBI in humans. Materials and Methods Docking studies were carried out to check the binding affinity of the drug toward the receptors. Various oxidative stress parameters, catalase level, glutathione level, superoxide dismutase (SOD) level, malondialdehyde (MDA), and nitric oxide levels, were measured. The mortality index and neuroprotective potential were carried out in TBI in D. melanogaster models. Results In the current study, there was an increase in oxidative stress following TBI as evidenced by a significant decrease in the catalase, glutathione, and SOD levels and increase in the level of MDA and nitric oxide after 24 hours. Antioxidant enzymes, catalase and glutathione peroxidase, have a dominant role in TBI. Docking studies were carried out on estrogen receptor 1 (pdb: 1TVO and 1UOM) and NDMA receptor (pdb: 3QEL) as agonist showing the binding affinity of the drug toward the receptors. In comparison to the vehicle-treated group, there was a dose-dependent significant increase in the SOD level and percentage climbing along with a decrease in the MDA level and total protein content. The mortality index was also observed at three concentrations of ranolazine (1, 2, and 4 mg/mL) in D. melanogaster homogenate. These findings suggest that ranolazine has a good neuroprotective potential in the treatment of TBI in the D. melanogaster model. Conclusion Present study concluded the scientific evaluation of neuroprotective potential of ranolazine in the treatment of TBI in the D. melanogaster model.

Morphofunctional Features of Glomeruli and Nephrons After Exposure to Electrons at Different Doses: Oxidative Stress, Inflammation, Apoptosis.

Kidney disease has emerged as a significant global health issue, projected to become the fifth-leading cause of years of life lost by 2040. The kidneys, being highly radiosensitive, are vulnerable to damage from various forms of radiation, including gamma (γ) and X-rays. However, the effects of electron radiation on renal tissues remain poorly understood. Given the localized energy deposition of electron beams, this study seeks to investigate the dose-dependent morphological and molecular changes in the kidneys following electron irradiation, aiming to address the gap in knowledge regarding its impact on renal structures. The primary aim of this study is to conduct a detailed morphological and molecular analysis of the kidneys following localized electron irradiation at different doses, to better understand the dose-dependent effects on renal tissue structure and function in an experimental model. Male Wistar rats (n = 75) were divided into five groups, including a control group and four experimental groups receiving 2, 4, 6, or 8 Gray (Gy) of localized electron irradiation to the kidneys. Biochemical markers of inflammation (interleukin-1 beta [IL-1β], interleukin-6 [IL-6], interleukin-10 [IL-10], tumor necrosis factor-alpha [TNF-α]) and oxidative stress (malondialdehyde [MDA], superoxide dismutase [SOD], glutathione [GSH]) were measured, and morphological changes were assessed using histological and immunohistochemical techniques (TUNEL assay, caspase-3). The study revealed a significant dose-dependent increase in oxidative stress, inflammation, and renal tissue damage. Higher doses of irradiation resulted in increased apoptosis, early stages of fibrosis (at high doses), and morphological changes in renal tissue. This study highlights the dose-dependent effects of electrons on renal structures, emphasizing the need for careful consideration of the dosage in clinical use to minimize adverse effects on renal function.

History of Preoperative Use of Gabapentin Before Lower Extremity Bypass Predisposes Patients to a High Risk of Opioid Use and Dependence in a Dose-Dependent Manner

IntroductionThis study assesses the association of preoperative use of gabapentinoids (GBPs) with postoperative risk of opioid-related disorders in peripheral artery disease patients undergoing lower extremity bypass operation. MethodsThis is a retrospective propensity score-matched analysis of patients undergoing peripheral artery bypass in TriNetX, a multicenter national database. Two study groups were constituted based on the preoperative history of prescribed GBPs. Primary outcomes were opioid-related disorders and mortality. The outcomes were reported at two-time endpoints that is, at 1 and 5 y. ResultsThis study population included a total of 23,706 patients. After propensity score-matched analysis, each group contained 5130 patients. The primary outcomes showed a significant increase in postoperative opioid-related disorders at the 1 and 5-y time points between GBPs and no GBPs groups: 1-y outcome (2.0% versus 1.1%; adj. P = 0.007) and 5-y outcome (4.5% versus 3.5%; adj. P = 0.035). Logistic regression analysis revealed an increase in the 1-y (adjusted odds ratio= 1.664; 95% CI [1.217, 2.273], P = 0.001) and 5-y (OR = 1.353; 95% CI [1.107, 1.653], P = 0.003) odds of opioid-related disorders in patients on GBPs. A secondary analysis showed a significant dose-dependent increase in the associated risk of 5-y opioid-related disorders in patients with a history of prescribed gabapentin. ConclusionsIn patients undergoing lower extremity bypass with a history of gabapentin use, there is an associated increased long-term risk of opioid-related disorders in a dose-dependent fashion. Overall, this study highlights weighing risks and benefits of prescribing GBPs for pain control versus their long-term associated risk of opioid-related disorders among other adverse outcomes.

DNA Damage, Cell Death, and Alteration of Cell Proliferation Insights Caused by Copper Oxide Nanoparticles Using a Plant-Based Model.

The speedy growth of copper oxide nanoparticle (CuO NP) manufacturing due to their wide application in industries has caused concerns due to their increased discharge into the environment from both purposeful and accidental sources. Their presence at an elevated concentration in the environment can cause potential hazards to the plant kingdom, specifically to staple food crops. However, limited research is available to determine the consequences of CuO NPs. The present study aimed to assess the morphological and cytological changes induced by CuO NPs on Pisum sativum L., a key staple food crop. Seeds of Pisum sativum were exposed to various concentrations of CuO NPs (0, 25, 50, 75, 100, and 125 ppm) for 2 h, and their effects on seed germination (SG), radicle length (RL), cell proliferation kinetics (CPK), mitotic index (MI), cell death (CD), micronucleus frequency (MNF), and chromosomal aberration frequency (CAF) were studied. The results indicate a significant reduction in SG, RL, CPK, and MI and a significant dose-dependent increase in CD, MNF, and CAF. CuO NP treatment has led to abnormal meiotic cell division, increased incidence of micronucleus frequency, and chromosomal aberration frequency. Additionally, the CuO NP-treated groups showed an increase in the percentage of aberrant meiotic cells such as laggard (LG), double bridge (DB), stickiness (STC), clumped nuclei (CNi), precocious separation (PS), single bridge (SB), and secondary association (SA). CuO NP treatment led to reductions in SG as follows: 55% at 24 h, 60.10% at 48 h, and 65% at 72 h; reductions in RL as follows: 0.55 ± 0.021 cm at 24 h, 0.67 ± 0.01 cm at 48 h, and 0.99 ± 0.02 cm at 72 h; reductions in CPK as follows: 34.98% at prophase, 7.90% at metaphase, 3.5% at anaphase, and 0.97% at telophase. It also led to a 57.45% increase in CD, a 39.87% reduction in MI, and a 60.77% increase in MNF at a higher concentration of 125 ppm. The findings of this study clearly show that CuO NPs have a genotoxic effect on the food crop plant Pisum sativum.

Toxicological evaluation of a herbal tea made from the leaves of Setaria megaphylla, Ageratum conyzoides and Chromolaena odorata

The toxicological effect of an herbal tea made from leaf extracts of Ageratum conyzoides, Chromolaena odorata, and Setaria megaphylla, plants traditionally used in Nigeria for malaria treatment was studied. Hot aqueous extracts of the ternary combination (1:1:1 ratio) of the plants were administered in varying doses (0, 50, 100, and 200 mg/kg) to assess impact on hepatic, renal and haematologic function parameters, and liver and kidney tissue histology of forty male albino rats divided into four groups of ten animals each. The tea extracts were administered orally daily for 60 days. Biochemical analyses revealed that the extract significantly (p&lt;0.05) reduced ALT (25.91 ± 2.35 to 6.84 ± 0.31 U/L), AST (36.39 ± 1.96 to 19.24 ± 1.19 U/L), and ALP (21.94 ± 3.51 to 11.06 ± 2.76 IU/L) activities, with no significant (p&gt;0.05) changes in bilirubin, protein and albumin concentrations, suggesting non-hepatotoxic property. Among the kidney function, significant (p&lt;0.05) dose-dependent increases was observed only in serum urea ranging from 23.09 ± 0.79 mg/dl (Control group) to 34.90 ± 0.52 mg/dl (200 mg/kg treated group). Increase in WBC count was observed indicative of a possible immune response, while other blood parameters remained stable. Histopathological examination confirmed that lower extract doses were relatively safe, whereas higher doses presented early signs of organ stress. These findings suggest that this herbal tea extract has a favourable safety profile at moderate doses, though higher doses may pose risk of organ toxicity, warranting further studies on dose optimization and elucidation of its folkloric therapeutic potential.

Phenylephrine Enhances the Mitogenic Effect of S-Allyl-L-cysteine on Primary Cultured Hepatocytes through Protein Kinase C-Induced B-Raf Phosphorylation.

The co-mitogenic effects of the α1-adrenoceptor agonist phenylephrine on S-allyl-L-cysteine (SAC)-induced hepatocyte proliferation were examined in primary cultures of adult rat hepatocytes. The combination of phenylephrine (10-10-10-6 M) and SAC (10-6 M) exhibited a significant dose-dependent increase in the number of hepatocyte nuclei and viable cells compared to SAC alone. This combination also increased the progression of hepatocyte nuclei into the S-phase. The potentiating effect of phenylephrine on SAC-induced cell proliferation was counteracted by prazosin (an α1-adrenergic receptor antagonist) and GF109203X (selective protein kinase C (PKC) inhibitor). In addition, PMA (direct PKC activator) potentiated the proliferative effects of SAC similarly to phenylephrine. In essence, these findings suggest that PKC activity plays a crucial role in enhancing SAC-induced cell proliferation. Moreover, the effects of phenylephrine on SAC-induced Ras activity, Raf phosphorylation, and extracellular signal-regulated kinase 2 (ERK2) phosphorylation were investigated. Phenylephrine (or PMA) in combination with SAC did not augment Ras activity, but further increased ERK2 phosphorylation and its upstream B-Raf phosphorylation. These results indicate that PKC activation, triggered by stimulating adrenergic α1 receptors, further amplifies SAC-induced cell proliferation through enhanced ERK2 phosphorylation via increased B-Raf-specific phosphorylation in primary cultured hepatocytes.

In vitro toxicity assessment of bioavailable iron in coal varieties of Central India

Introduction Information on bioavailable Iron (BAI) content in respirable coal dust (RCD) is crucial to address occupational health and safety, especially in preventing coal workers’ pneumoconiosis (CWP). Materials and methods In the present study, we determined BAI concentrations in seventy-seven coal samples collected from ten coal mining regions of Central India. The cytotoxic potential of BAI-RCD was established invitro by using alveolar epithelial (A549) and macrophage (U937) cell lines. The oxidative/antioxidant status, inflammations, and genotoxicity attributed to BAI-RCD exposure were evaluated and correlated with CWP pathophysiology. Results The mean BAI concentrations in the coal samples (n = 77) range from (275 to 9065 mg kg-1) and showed wide variability. Both cell lines were exposed to low (275 mg kg-1), moderate (4650 mg kg-1), and high (9065 mg kg-1) BAI-RCD samples showed significant (p &lt; 0.001) cytotoxicity in a dose-dependent manner (low &lt; moderate &lt; high) compared to the control. After BAI-RCD treatment, both cell lines showed a decrease in antioxidant stress measures (SOD, CAT, and GSH) and a significant (p &lt; 0.001) increase in oxidative stress parameters (NADPH, MPO, LPO, and PC). Furthermore, these cell line models demonstrated a statistically significant (p &lt; 0.001) dose-dependent increase in cytokines (TGF-β1, IL-1β, TNF-α, MCP-1, and IL-6 cytokines) and oxidative DNA damage marker (8-OH-dG). Conclusion Results indicated that the central India coals (even at low BAI content) may be accountable for inflammatory responses and cytotoxicity. Hence, BAI can be important characteristic to establish safety standards for coal dust exposure before active mining.

Evaluation of Larvicidal Properties of Zingiber officinale Rhizome and Allium Sativum Bulbs against Aedes aegypti

This study evaluates the larvicidal potential of methanolic extracts derived from Zingiber officinale (ginger) rhizome and Allium sativum (garlic) bulbs against the 4th instar larvae of Aedes aegypti, a key vector responsible for transmitting diseases such as dengue, yellow fever, chikungunya, and Zika. Due to the increasing resistance of mosquitoes to conventional insecticides, there is a pressing need for alternative, eco-friendly vector control methods. Botanical extracts, known for their pesticidal properties, offer a promising solution. Methanolic extracts were collected from the selected test plant parts and different test solutions were prepared to test their larvicidal efficacy against the fourth instar larvae of Ae. aegypti. The results indicated a significant dose-dependent increase in larval mortality for both extracts. The extracts of Z. officinale consistently showed better larvicidal effects than outperformed the extracts of A. sativum at all tested concentrations, with mortality rates ranging from 14.29% at 62.5 ppm to 79.22% at 500 ppm. In contrast, A. sativum exhibited mortality rates from 11.69% at 62.5 ppm to 76.62% at 500 ppm. The LC50 values further highlighted the greater efficacy of ginger extract, with an LC50 of 219.10 ppm compared to 237.39 ppm for garlic. These results suggest the usage of extracts of ginger rhizomes and garlic bulbs as eco-friendly larvicides against Ae. aegypti mosquitoes.

Promoting effect of sunset yellow on N-methyl N-nitrosourea-induced rat mammary carcinogenesis: Implications of molecular mechanisms

BackgroundNowadays, the use of food additives, such as Sunset Yellow (SY), is growing, which attracted attention to the potential relationship between some diseases and food additives. AimThe study aimed to investigate the role of Sunset Yellow during chemically-induced mammary gland carcinogenesis in Sprague–Dawley rats. Material and methodsThree groups of female rats were intraperitoneally administered with N-methyl-N-nitrosourea (MNU). Group 1 was set on a basal diet. Group 2 was treated with 161.4 mg\\kg\\day Sunset Yellow (SY). Group 3 was given SY at 80.7 mg\\kg\\day. Groups 4–6 were not administered MNU; Group 4 received vehicles only. Groups 5 and 6 were administered SY similarly to groups 2 and 3 respectively. ResultsSunset Yellow at both doses exerted a significant dose-dependent increase in tumor incidences, multiplicities, volumes, and decreased tumor latency as compared with control. Immunolabeling indexes of the proliferating cell nuclear antigen, estrogen receptor alpha, and progesterone receptor were significantly increased after SY treatment. Oxidative stress markers, serum estrogen, progesterone, and prolactin levels were significantly modified by SY treatment. The mRNA expression of estrogen receptor alpha and epidermal growth factor was up-regulated in SY groups versus control. ConclusionCollectively, SY has significantly promoted MNU-induced mammary tumors in rats with underlying mechanisms correlating SY consumption with estrogen disruption and subsequent antioxidative stress discrepancy.

Investigation of [11C]carfentanil for mu opioid receptor quantification in the rat brain

[11C]Carfentanil ([11C]CFN) is the only selective carbon-11 labeled radiotracer currently available for positron emission tomography (PET) imaging of mu opioid receptors (MORs). Though used extensively in clinical research, [11C]CFN has not been thoroughly characterized as a tool for preclinical PET imaging. As we were occasionally observing severe vital sign instability in rat [11C]CFN studies, we set out to investigate physiological effects of CFN mass and to explore its influence on MOR quantification. In anesthetized rats (n = 15), significant dose-dependent PCO2 increases and heart rate decreases were observed at a conventional tracer dose range (IV, > 100 ng/kg). Next, we conducted baseline and retest [11C]CFN PET scans over a wide range of molar activities. Baseline [11C]CFN PET studies (n = 27) found that nondisplaceable binding potential (BPND) in the thalamus was positively correlated to CFN injected mass, demonstrating increase of MOR availability at higher injected CFN mass. Consistently, when CFN injected mass was constrained < 40 ng/kg (~ 10% MOR occupancy in rats), baseline MOR availability was significantly decreased. For test–retest variability (TRTV), better reproducibility was achieved by controlling CFN injected mass to limit the difference between scans. Taken together, we report significant cardiorespiratory depression and a paradoxical influence on baseline MOR availability at conventional tracer doses in rats. Our findings might reflect changes in cerebral blood flow, changes in receptor affinity, or receptor internalization, and merits further mechanistic investigation. In conclusion, rat [11C]CFN PET requires stringent quality assurance of radiotracer synthesis and mass injected to avoid pharmacological effects and limit potential influences on MOR quantification and reproducibility.

Abstract A013: NF1 loss is syntetic lethal with Trastuzumab emtansine

Abstract There is great interest in the identification of biomarkers to guide development of antibody-drug conjugates (ADC). Most research has focused on target expression, but key predictors of payload efficacy have not been indeitifed. NF1 is a tumor suppressor classically considered as an inhibitor of RAS signaling, and often mutated in metastatic HER2+ breast cancer (BC). We screened multiple approved drugs for differential sensitivityin CRISPR-engineeredf NF1 KO cells. HER2-targeted agents (small molecules or antibodies) were found to be less effective upon NF1 loss; surprisingly, we identified increased sensitivity to the approved ADC T-DM1, but not to Trastuzumab Deruxtecan (T-Dxd). We then elucidated the underlying molecular cause employing in vivo, in vitro and in vitro reductionist systems. To measure in vivo MT dynamics, we transiently transfected the MT end-binding protein EB3- GFP and reconstructed MT trajectories by live-cell imaging. Upon DM1 treatment, KO cells showed a highly significant reduction in MT speed, demonstrating a direct role for NF1 on MT dynamics in cells. In tubulin polymerization assays, recombinant NF1 greatly accelerated polymerization and completely rescued DM1-induced inhibition. NF1 induced significant MT bundling, a defining feature of many MT-associated proteins, which generates signal indistinguishable from true MT polymerization in turbidity assays. To follow the dynamics of individual microtubules, we applied Total Internal Reflection (TIRF) microscopy on glass-immobilized MTs. As expected, polymerization in the presence of NF1 led to a dose-dependent significant increase in MT dynamics (fraction of elongating MTs, elongation speed, catastrophe rate). Expectedly, DM1 led to significant reduction in the fraction of elongating MTs and speed, but these defects were completely or partially rescued by NF1. Importantly, DM1 did not only lead to MT shortening (as proposed by the current model), but also to clear and frequent MT fracturing, indicating that the drug is not only engaging MT ends but also intra-tubular binding sites. This is consistent with recent models of MT formation which incorporate the frequent presence of areas of discontinuity or damage induced by mechanical stress, exposing intratubular DM1 binding sites. Interestingly, adding NF1 to DM1-treated MTs generated areas of de novo intra-tubular tubulin insertion, coincident with damaged sites, suggesting an entirely novel role for NF1 in MT repair. In conclusion, we provide evidence for a model in which maytansinoids bind not only to soluble tubulin dimers and MT ends, but also to intratubular damaged sites. Thus, the number of binding sites in cells would be proportional to MT damage, suggesting a mechanism for differential efficacy across tumor types and a potential avenue for combinatorial drug development. These results prompt the use of NF1 as a biomarker to select patients for ADC treatment. Funding: FIEO fellowship 2023, AIRC (n25791), Italian MoH-Ricerca Corrente di Rete (ACCORD) 2022, Next Generation EU – PNRR M6C2 – PNRR-MAD-2022-12376934 Citation Format: Eleonora Messuti, Bruno Achutti Duso, Alessia Castiglioni, Giulia Tini, Emanuele Bonetti, Giuseppe Ciossani, Silvia Monzani, Daria Khuntsariya, Marcus Braun, Zdenek Lansky, Luigi Scietti, Luca Mazzarella. NF1 loss is syntetic lethal with Trastuzumab emtansine [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Expanding and Translating Cancer Synthetic Vulnerabilities; 2024 Jun 10-13; Montreal, Quebec, Canada. Philadelphia (PA): AACR; Mol Cancer Ther 2024;23(6 Suppl):Abstract nr A013.

A rare olive compound oleacein functions as a TrkB agonist and mitigates neuroinflammation both in vitro and in vivo

BackgroundNeuroinflammation is widely acknowledged as a characteristic feature of almost all neurological disorders and specifically in depression- and anxiety-like disorders. In recent years, there has been significant attention on natural compounds with potent anti-inflammatory effects due to their potential in mitigating neuroinflammation and neuroplasticity.MethodsIn the present study, we aimed to evaluate the neuroprotective effects of oleacein (OC), a rare secoiridoid derivative found in extra virgin olive oil. Our goal was to explore the BDNF/TrkB neurotrophic activity of OC and subsequently assess its potential for modulating neuroinflammatory response using human neuroblastoma cells (SH-SY5Y cells) and an in vivo model of depression induced by lipopolysaccharide (LPS)-mediated inflammation.ResultsIn SH-SY5Y cells, OC exhibited a significant dose-dependent increase in BDNF expression. This enhancement was absent when cells were co-treated with inhibitors of BDNF's receptor TrkB, as well as downstream molecules PI3K and MEK. Whole-transcriptomics analysis revealed that OC upregulated cell cycle-related genes under normal conditions, while downregulating inflammation-associated genes in LPS-induced conditions. Furthermore, surface plasmon resonance (SPR) assays demonstrated that OC exhibited a stronger and more stable binding affinity to TrkB compared to the positive control, 7,8-dihydroxyflavone.Importantly, bioluminescence imaging revealed that a single oral dose of OC significantly increased BDNF expression in the brains of Bdnf-IRES-AkaLuc mice. Furthermore, oral administration of OC at a dosage of 10 mg/kg body weight for 10 days significantly reduced immobility time in the tail suspension test compared to the LPS-treated group. RT-qPCR analysis revealed that OC significantly decreased the expression of pro-inflammatory cytokines Tnfα, Il6, and Il1β, while simultaneously enhancing Bdnf expression, as well as both pro and mature BDNF protein levels in mice hippocampus. These changes were comparable to those induced by the positive control antidepressant drug fluoxetine. Additionally, microarray analysis of mouse brains confirmed that OC could counteract LPS-induced inflammatory biological events.ConclusionAltogether, our study represents the first report on the potential antineuroinflammatory and antidepressant properties of OC via modulation of BDNF/TrkB neurotrophic activity. This finding underscores the potential of OC as a natural therapeutic agent for depression- and anxiety-related disorders.

Kartogenin Induces Chondrogenesis in Cartilage Progenitor Cells and Attenuates Cell Hypertrophy in Marrow-Derived Stromal Cells.

Kartogenin (KGN) is a synthetic small molecule that stimulates chondrogenic cellular differentiation by activating smad-4/5 pathways. KGN has been proposed as a feasible alternative to expensive biologic growth factors, such as transforming growth factor β, which remain under strict regulatory scrutiny when it comes to use in patients. This study reports the previously unexplored effects of KGN stimulation on cartilage- derived mesenchymal progenitor cells (CPCs), which have been shown to be effective in applications of cell-based musculoskeletal tissue regeneration. Our findings demonstrate that KGN treatment significantly increased markers of chondrogenesis, SOX9 and COL2 following 3-10 days of treatment in human CPCs. KGN treatment also resulted in a significant dose-dependent increase in GAG production in CPCs. The same efficacy was not observed in human marrow-derived stromal cells (BM-MSCs); however, KGN significantly reduced mRNA expression of cell hypertrophy markers, COL10 and MMP13, in BM-MSCs. Parallel to these mRNA expression results, KGN led to a significant decrease in protein levels of MMP-13 both at 0-5 days and 5-10 days following KGN treatment. In conclusion, this study demonstrates that KGN can boost the chondrogenicity of CPCs and inhibit hypertrophic terminal differentiation of BM-MSCs.

Dose-dependent LSD effects on cortical/thalamic and cerebellar activity: brain oxygen level-dependent fMRI study in awake rats.

Lysergic acid diethylamide is a hallucinogen with complex neurobiological and behavioural effects. This is the first study to use MRI to follow functional changes in brain activity in response to different doses of lysergic acid diethylamide in fully awake, drug-naive rats. We hypothesized that lysergic acid diethylamide would show a dose-dependent increase in activity in the prefrontal cortex and thalamus while decreasing hippocampal activity. Female and male rats were given intraperitoneal injections of vehicle or lysergic acid diethylamide in doses of 10 or 100 µg/kg while fully awake during the imaging session. Changes in blood oxygen level-dependent signal were recorded over a 30-min window. Approximately 45-min post-injection data for resting-state functional connectivity were collected. All data were registered to rat 3D MRI atlas with 173 brain regions providing site-specific increases and decreases in global brain activity and changes in functional connectivity. Treatment with lysergic acid diethylamide resulted in a significant dose-dependent increase in negative blood oxygen level-dependent signal. The areas most affected were the primary olfactory system, prefrontal cortex, thalamus and hippocampus. This was observed in both the number of voxels affected in these brains regions and the changes in blood oxygen level-dependent signal over time. However, there was a significant increase in functional connectivity between the thalamus and somatosensory cortex and the cerebellar nuclei and the surrounding brainstem areas. Contrary to our hypothesis, there was an acute dose-dependent increase in negative blood oxygen level-dependent signal that can be interpreted as a decrease in brain activity, a finding that agrees with much of the behavioural data from preclinical studies. The enhanced connectivity between thalamus and sensorimotor cortices is consistent with the human literature looking at lysergic acid diethylamide treatments in healthy human volunteers. The unexpected finding that lysergic acid diethylamide enhances connectivity to the cerebellar nuclei raises an interesting question concerning the role of this brain region in the psychotomimetic effects of hallucinogens.

Https://www.texilajournal.com/public-health/article/2527-guardians-of-liver

Glyphosate is used as an herbicide in agriculture. At sub-agriculture concentrations, glyphosate-based herbicide inhibits cell proliferation. Glyphosate is a chelating agent that interferes with the metabolic activities in plants thereby adversely affecting its metabolism. The study aimed to determine the glyphosate-induced detrimental changes in SREBP-1c and PPAR-γ mRNA expression in adipose tissue of adult male rats. Adult male Wistar albino rats were divided into 4 groups, each consisting of 6 animals. Group I served as normal control rats; Group II-IV consisted of rats exposed to glyphosate at different concentrations (50, 100, and 250 mg/kg body weight respectively) orally for 16 weeks. After 16 weeks of treatment, the animals were sacrificed, and adipose tissue was dissected out for the assessment of SREBP-1c and PPAR-γ mRNA by real-time PCR using gene-specific down-regulated primers. The results with the p&lt;0.05 level were considered to be statistically significant. The results showed a significant dose-dependent increase (P &lt;0.05) in the expression of SREBP-1c in all the glyphosate-exposed rats compared to control rats and PPAR-γ mRNA expression was found to be significantly reduced in a concentration-dependent manner (P&lt;0.05) compared to normal control animals. The current findings for the first time report that glyphosate had detrimental changes in the expression of transcription factors such as SREBP-1c and PPAR-γ mRNA in adipose tissue and thereby glyphosate may lead to the development of type-2 diabetes or insulin resistance.