Intraperitoneal Dose Research Articles

A phase I trial of intraperitoneal (IP) mesothelin (MSLN)-targeted CAR T-cell therapy in patients with MSLN-positive esophagogastric cancer (EGC) with peritoneal carcinomatosis.

TPS513 Background: Outcomes for patients with advanced EGC remain poor. The peritoneum is an immune-privileged niche and a common site of metastases occurring in up to 40% of patients with metastatic EGC; this is characterised by poor survival because of resistance to immunotherapy and a lack of measurable disease, which excludes trial participation. MSLN was selected as a target for CAR T cell therapy as it is overexpressed in up to 60% of EGC, is associated with aggressive disease biology and has low levels of expression in normal tissues. In pre-clinical in vivo models of peritoneal carcinomatosis, improved efficacy was observed with regional administration of IP CAR T cells, compared with systemic CAR T cells. No significant toxicity was observed at clinically relevant doses of IP administered CAR T-cells. M28z1XXPD1DNR CAR T-cells are a next-generation MSLN-targeted CAR, equipped with a modified CD3z (1XX), and a PD-1 dominant negative receptor (PD1DNR) that provides T-cell intrinsic checkpoint blockade. Methods: This is a single-arm phase I study assessing the safety, maximum tolerated dose (MTD) and preliminary efficacy of IP administered M28zXXPD1DNR CAR T cells in patients with MSLN-positive (IHC ≥ 25%) EGC with evidence of peritoneal carcinomatosis (imaging and/or cytology). Patients with concomitant extraperitoneal disease are eligible. Patients must have received at least one prior line of treatment for advanced or metastatic disease and have at least one measurable or evaluable lesion per RECIST 1.1. A minimum of 4 patients and maximum of 18 patients will be treated with 4 escalating doses (with 1 “fallback” dose) of CAR T cells to a maximum of 3 x 10 7 cells/kg administered via peritoneal catheter (Table). Lymphodepletion with fludarabine 30mg/m 2 /day and cyclophosphamide 300mg/m 2 /day is given for 3 days prior to the CAR T cell infusion. Adverse events, response rates (RECIST 1.1), survival outcomes and immuno-monitoring (immune cell phenotyping and serum/peritoneal fluid cytokine analysis) will be assessed. The trial is currently enrolling patients. Clinical trial information: Will be registered before ASCO GI. Dose-escalation scheme based on the continuous reassessment method. Dose Level Dose (cells/kg) 1 1 x 10 6 2 (Starting dose) 3 x 10 6 3 6 x 10 6 4 1 x 10 7 5 3 x 10 7

Luteolin alleviates diabetic cardiac injury related to inhibiting SHP2/STAT3 pathway.

Diabetic cardiomyopathy, a heart disease resulting from diabetes mellitus, inflicts structural and functional damage to the heart. Recent studies have highlighted the potential role of luteolin, a flavonoid, in mitigating diabetic cardiovascular injuries. The Src homology 2-containing protein tyrosine phosphatase 2 (SHP2) is implicated in exacerbating diabetes- and obesity-related complications. Interestingly, luteolin has been shown to inhibit protein tyrosine phosphatases, but it's unclear how SHP2 relates to luteolin's protective effects against diabetic heart disease. Here, we hypothesized that the inhibition of SHP2 signaling could play a role in luteolin's protective action against diabetic heart injury. Diabetes was induced in male Sprague-Dawley rats through a high-fat diet followed by a single intraperitoneal dose of streptozotocin (30mg/kg). Five weeks post-diabetes induction, these rats were intraperitoneally injected with luteolin at varying doses (5, 10, 20mg/kg) every other day for an additional 5 weeks. Then cardiac function was assessed, and hearts were isolated for further analysis. We found that luteolin notably improved cardiac function, inhibited cardiac hypertrophy and fibrosis, reduced levels of inflammatory factors and reactive oxygen species, and activated superoxide dismutase. Importantly, luteolin treatment also reduced the expression of SHP2 and phosphorylated signal transducer and activator of transcription 3 (STAT3) in a dose-dependent manner. These findings suggest that luteolin protects the diabetic heart against inflammation, oxidative stress, hypertrophy, and fibrosis, which may relate to down-regulating cardiac SHP2/STAT3 signaling.

Effect of methanol extract of cocos nucifera husk (MECH) in Alloxan-induced toxicity in Wistar male rats

The present study investigated the effect of Methanol extract of Cocos nucifera Husk (MECH) in alloxan-induced toxicity in male wistar rats. Thirty-six (36) male rats were randomly divided into six experimental groups (n=6). Group 1 was administered distilled water). Groups 2, 3, 4, 5and 6 were made diabetic by a single intraperitoneal dose of 150mg/kg body weight (bwt) of alloxon monohydrate. Group 2 served as diabetic non-treated, Group 3 (positive control) was treated orally with gibenclamide at 2mg/kg bwt. Groups 4, 5, and 6 were treated with MECH extract orally at doses of 62.5mg/kg, 125.0mg/kg, and 250mg/kg body weight for 28 days respectively. The fasting blood sugar (FBS) was taken on days 0, 1, 7, 14, 21, and 28. At the end of 28 days, the serum biochemical parameters were measured (assigned). MECH demonstrated a significant (P<0.05) dose-dependent reduction in the fasting blood sugar (FBS) of the diabetic rats as compared to the untreated control group. A decrease of 68.5%, 78.25%, and 47% respectively on day 28. In other words, the highest reduction was seen at 125mg/kg FBS. There was a significant (P<0.05) increase in levels of Alanine aminotransferase (ALT), Alkaline phosphatase (ALP) and urea in the alloxan-intoxicated rat when compared with normal control. MECH reduced the level of ALT, ALP, and urea compared with the diabetic control. These findings showed that MECH has significant antidiabetic activity where comparable with glibenclamide. It also may protect against DM-induced hepatorenal injuries in rats.

Therapeutic Potential of Asparagus Racemosus Root in Managing Hyperglycemia, Dyslipidemia, and Platelet Aggregation in Neonatal Streptozotocin-induced Type 2 Diabetic Rats

Lipid abnormalities frequently accompany hyperglycemia and thus a primary goal in diabetes therapy is the management of dyslipidemia. Asparagus racemosus root has previously been shown to reduce postprandial blood glucose in diabetic rats by delaying carbohydrate absorption and enhancing insulin secretion. In the present study, the chronic effects of A racemosus root on serum glucose, fructosamine, lipids, and platelet aggregation were assessed in rats with type 2 diabetes induced by neonatal streptozotocin injection. The type 2 diabetes model was created by injecting 48-hour-old pups with a single intraperitoneal dose of streptozotocin (STZ). Platelet aggregation was measured by optical aggregometry. Daily oral administration of ethanol extract of A racemosus to diabetic rats (n = 10) lowered serum glucose by 21% (p<0.01) and fructosamine by 11% (p<0.05) after 28 days. Total cholesterol (p<0.05), triglyceride (p<0.05), and NEFA (p<0.01) levels were also lowered by 9%, 16% and 38% respectively. No difference in HDL cholesterol or body weights was observed compared to control rats but platelet aggregation was significantly reduced by 18% (p<0.05). Food and water intake, stool formation, water content of stools, and urine formation were unchanged in extract-treated rats in a 24-hour acute observational study in Nalgene Metabolic Cages. In conclusion, this study reveals that ethanol extract of A racemosus root lowers circulating glucose, and atherogenic blood lipids and decreases platelet aggregation. Thus, A racemosus is the source of glucose-lowering bioactive agents and a useful dietary adjunct in the management of diabetes, dyslipidemia, and related complications.

The role of Juniperus Macrocarpa extract as anti-inflammatory and antioxidant on methotrexate-induced acute liver injury in rat model

Background Methotrexate (MTX) is an antifolate medication indicated to treat an array of tumors and autoinflammatory maladies. MTX may exhibit harmful impacts on multiple organs, especially liver injury and cirrhosis. Juniperus macrocarpa is a medicinal herb enriched with polyphenols and flavonoids featuring robust anti-inflammatory and antioxidative benefits. Objective To evaluate the hepatoprotective effects of Juniperus macrocarpa aqueous extract on MTX-aggravated liver toxicity. Methods The study involved 20 male middle-aged albino rats, arbitrarily allocated into 4 groups of 5 animals each. Group 1 (control) were given distilled water (DW) once daily for two weeks. Group 2 (MTX) got an intraperitoneal single dose of MTX (20 mg/kg) for two weeks. Rats in groups 3 and 4 were given daily dosages of 100 mg and 200 mg of Juniperus macrocarpa aqueous extract, respectively, for two weeks before receiving a single intraperitoneal MTX injection. Results Juniperus macrocarpa extracts at both low and high doses substantially alleviated the MTX-provoked biochemical alterations, as evidenced by decreased levels of inflammatory parameters including TNF-α and IL-6 and hepatic enzymes including ALT, AST, and ALP. Juniperus macrocarpa also significantly boosted levels of the anti-oxidant enzymes like SOD and GPX. Moreover, Juniperus macrocarpa extract attenuated congestive and degenerative hepatic changes, as indicated by improved histopathological findings. Conclusion The anti-oxidative and anti-inflammatory activities of Juniperus macrocarpa extract are a promising approach for ameliorating MTX-aggravated hepatotoxicity.

Lentinan's effect on gut microbiota and inflammatory cytokines in 5-FU-induced mucositis mice

Chemotherapeutic therapies for cancer are frequently associated with cytotoxic side effects that can be harmful to human health, including the development of intestinal mucositis (IM). It mostly affects the gastrointestinal tract, causing ulceration, inflammation, and the formation of lesions in the colon. Surprisingly, despite the frequency of IM, therapeutic choices remain restricted. In our search for new intestinal mucositis therapies, we wanted to see how Lentinan (LT), derived from Lentinus edodes, would fare in mouse models of intestinal mucositis. To create the intestinal mucositis model in mice, we gave them intra-peritoneal doses of 5-fluorouracil (5-FU) (50 mg/kg) and then tested the effects of Lentinan on intestinal mucositis. This examination required constant monitoring of several factors, such as body weight fluctuations, food consumption, and diarrhea. In addition, we measured the levels of certain inflammatory cytokines (Tumour Necrosis Factor-alpha (TNF-α), Interleukin-1 (IL-1), Interleukin-6 (IL-6), and Interleukin-10 (IL-10), looked at the expression of tight junction proteins (Zonula Occludens-1(ZO-1), Claudin-1), measured mucin-2 levels, and looked into changes in the gut flora. In the mouse model of intestinal mucositis, our findings showed that LT effectively reduced weight loss, increased food intake, and relieved diarrhea. Concurrently, we saw a decrease in the expression of inflammatory cytokines such as TNF-α, IL-1, and IL-6, as well as a considerable increase in the concentration of IL-10. Furthermore, LT reduced intestinal mucositis by increasing the length and structural integrity of the colon. Furthermore, increased expression of tight junction proteins (ZO-1, Claudin-1), mucin-2, and an increase in the number of goblet cells all confirmed our previous findings. Notably, the makeup of beneficial bacteria in the stomach increased as well. Finally, our findings suggest that LT can effectively prevent 5-fluorouracil-induced intestinal mucositis in mice by improving immune function, restoring intestinal barrier integrity, and rebalancing gut microbial flora.

Protective effects of hydatid cyst fluid on inflammation and tissue damage in rat model of type 1 diabetes.

Cystic echinococcosis, caused by Echinococcus granulosus, is a zoonotic disease with immunomodulatory properties attributed to hydatid cyst fluid (HCF). Given the immune-modulating and anti-inflammatory properties of HCF observed in other contexts, its potential therapeutic effects in diabetes remain unexplored. This study aimed to investigate the potential therapeutic effects of HCF on glycemic control, inflammatory cytokines, and tissue histopathology in a streptozotocin (STZ)-induced model of type 1 diabetes. Twenty male rats were randomly divided into four groups (n = 5): a healthy control group, a hydatid cyst group that received three intraperitoneal injections of HCF at two-week intervals, a diabetic group that received a single intraperitoneal dose of STZ to induce diabetes, and a hydatid cyst + diabetic group (HCF + STZ) that received both HCF treatment and STZ administration. Serum glucose levels, inflammatory cytokines (TNF-α, IL-1β, and IL-10), and histopathological changes in pancreatic and renal tissues were analyzed. The HCF + STZ group demonstrated a significant reduction in serum glucose levels compared to the STZ-only group. Pro-inflammatory cytokines TNF-α and IL-1β were significantly decreased in HCF-treated diabetic rats, while the anti-inflammatory cytokine IL-10 was partially restored. Histopathological examination revealed severe pancreatic islet atrophy and renal degeneration in the diabetic group, which were markedly alleviated in the HCF + STZ group. These findings suggest that HCF's immunomodulatory and anti-inflammatory properties may mitigate hyperglycemia and inflammatory responses in type 1 diabetes, warranting further investigation into its mechanisms and clinical applications.

Improving the Safety of N,N -Dimethylacetamide (DMA) as a Potential Treatment for Preterm Birth in a Pregnant Mouse Model Using a Vaginal Nanoformulation.

Vaginal administration and the uterine first pass effect allow for preferential delivery of drugs to the reproductive tract. Dimethylacetamide has previously been shown to delay preterm birth in a pregnant mouse model when given intraperitoneally but the effectiveness of a vaginal nanoformulation of dimethylacetamide has yet to be tested. The purpose of this study was to compare the two formulations of dimethylacetamide for efficacy in rescuing pups from preterm birth in an inflammation-induced mouse model, effects on the maternal fetal interface, and pharmacokinetic profiles in maternal plasma. Timed pregnant CD1 mice were given a 1.56 mg/kg intraperitoneal dose of lipopolysaccharide followed by 3 doses of either vaginal dimethylacetamide or intraperitoneal dimethylacetamide. Mice were monitored for 48 hours and times of deliveries were recorded. Additionally, CD1 mice in late gestation were given a single dose of either vaginal or intraperitoneal dimethylacetamide and blood was drawn at 3 different time points following administration. Vaginal administration of dimethylacetamide had similar efficacy in delaying inflammation induced preterm birth as intraperitoneal administration but resulted in lower concentrations in the systemic circulation and decreased effects on the maternal fetal interface. Vaginal nanoformulations should be explored for their potential therapeutic value for the delay of preterm birth.

Exploring Azithromycin's Neuroprotective Role in Traumatic Brain Injury: Insights into Cognitive and Motor Recovery and Neuroinflammatory Modulation.

Traumatic brain injury (TBI) is a leading cause of mortality worldwide and often results in substantial cognitive, motor, and psychological impairments, triggering oxidative stress, neuroinflammation, and neurodegeneration. This study examined the neuroprotective effects of azithromycin (AZI) in TBI. TBI was induced in rats using the weight-drop method. Subsequently, rats received a daily intraperitoneal (I.P.) dose of AZI (150 mg/kg) for 28 days. Behavioral tests (Morris water maze, rotarod, and open field tests) were performed to assess cognitive and motor functions. Neurochemical analyses included oxidative stress markers (GSH, SOD, MDA, catalase), inflammatory cytokines (TNF-α, IL-1β), apoptotic markers (caspase-3, Bax, Bcl-2), mitochondrial complexes (complex I, II, III, IV, and V), and the transforming growth factor- beta (TGF-β) as a neurofilament marker. Histological evaluations focused on neuronal integrity in the cortex, hippocampus, and striatum. Treatment with AZI significantly facilitated motor and cognitive function recovery in TBI-affected rats. At the molecular level, AZI effectively reduced oxidative stress markers, ameliorated neuroinflammation by decreasing TNF-α, IL-1β, and neuronal apoptosis, and differentially modulated mitochondrial complexes. Histological assessments revealed enhanced neuronal integrity and fewer pathological changes in AZI-treated rats compared to untreated TBI controls. AZI was shown to interfere with several pathways involved in TBI's pathophysiology. While preclinical results are promising, further studies are necessary to establish the long-term safety and efficacy of AZI in a clinical setting. This research supports the potential re-purposing of AZI as a novel treatment strategy for TBI and related neurodegenerative disorders.

Possibility of Using NO Modulators for Pharmacocorrection of Endothelial Dysfunction After Prenatal Hypoxia.

Prenatal hypoxia (PH) is a key factor in the development of long-term cardiovascular disorders, which are caused by various mechanisms of endothelial dysfunction (ED), including those associated with NO deficiency. This emphasizes the potential of therapeutic agents with NO modulator properties, such as Thiotriazoline, Angiolin, Mildronate, and L-arginine, in the treatment of PH. Methods: Pregnant female rats were given a daily intraperitoneal dose of 50 mg/kg of sodium nitrite starting on the 16th day of pregnancy. A control group of pregnant rats received saline instead. The resulting offspring were divided into the following groups: Group 1-intact rats; Group 2-rat pups subjected to prenatal hypoxia (PH) and treated daily with physiological saline; and Groups 3 to 6-rat pups exposed to prenatal hypoxia and treated daily from the 1st to the 30th day after birth. Levels of sEPCR, Tie2 tyrosine kinase, VEGF-B, SOD1/Cu-Zn SOD, GPX4, and GPX1 in the heart's cytosolic homogenate were assessed using ELISA. The expression of VEGF and VEGF-B mRNA was analyzed via real-time polymerase chain reaction, and the nuclear area of myocardial microvessel endothelial cells was evaluated morphometrically. Results: We have shown that only two representatives of this group-Angiolin and Thiotriazoline-are able to exert full effect on the indices of endothelial dysfunction after PH to decrease sEPCR, increase Tie-2, VEGF-B and VEGF-B mRNA, Cu/ZnSOD, and GPX in myocardial cytosol, and increase the area of endotheliocyte nuclei in 1- and 2-month-old rats in comparison with the control. Conclusions: Our results experimentally substantiate the necessity of early postnatal cardio- and endothelioprotection using NO modulators, taking into account the role of NO-dependent mechanisms in the pathogenesis of cardiovascular system disorders in neonates after PH.

Protective Effects of Phycobiliproteins from Arthrospira maxima (Spirulina) Against Cyclophosphamide-Induced Embryotoxicity and Genotoxicity in Pregnant CD1 Mice.

Background/Objectives: In recent years the global incidence of cancer during pregnancy is rising, occurring in 1 out of every 1000 pregnancies. In this regard, the most used chemotherapy drugs to treat cancer are alkylating agents such as cyclophosphamide (Cp). Despite its great efficacy, has been associated with the production of oxidative stress and DNA damage, leading to embryotoxicity, genotoxicity, and teratogenicity in the developing conceptus. Therefore, this study aimed to investigate the protective role of phycobiliproteins (PBP) derived from Arthrospira maxima (spirulina) in reducing Cp-induced embryotoxicity and genotoxicity in pregnant CD1 mice. Methods: Pregnant CD1 mice were divided into five groups: control, Cp 20 mg/kg, and three doses of PBP (50, 100, and 200 mg/kg) + Cp co-treatment. PBP were administered orally from day 6 to 10.5 dpc, followed by a single intraperitoneal dose of Cp on 10.5 dpc. Embryos were collected at 12.5 dpc to assess morphological development and vascular alterations, while maternal DNA damage was evaluated using micronucleus assays and antioxidant enzyme activity in maternal plasma. Results: PBP exhibited a dose-dependent protective effect against Cp-induced damage. The 200 mg/kg PBP dose significantly reduced developmental abnormalities, micronucleated polychromatic erythrocytes, and oxidative stress, (as evidenced by increased SOD and GPx activity). Conclusions: Phycobiliproteins from Arthrospira maxima (spirulina) effectively reduced Cp-induced morphological and vascular alterations in embryos and genotoxicity in pregnant mice. These findings highlight their potential as a complementary therapy to mitigate teratogenic risks during chemotherapy. Further research is needed to optimize dosing and explore clinical applications.

Iron(III)-Quercetin Complex: In Vivo Acute Toxicity and Biodistribution of Novel MRI Agent.

The iron(III)-quercetin complex, known as "IronQ", is an innovative MRI contrast agent composed of one Fe(III) ion and two quercetin molecules. IronQ is efficiently internalized by cells, enabling T1-weighted MRI tracking. It has demonstrated therapeutic benefits in reducing inflammation in an intracerebral hemorrhage (ICH) mouse model and offers a safer alternative to gadolinium-based agents by avoiding cytotoxicity and genotoxicity. These properties make IronQ a promising candidate for safe and effective MRI contrast enhancement. This study aims to further the development of IronQ as an MRI contrast agent by investigating its biodistribution, pharmacokinetics, and acute toxicity in a preclinical animal model. The relaxivity of IronQ was measured in water and whole blood phantoms. Acute toxicity was evaluated in Sprague Dawley rats administered single intraperitoneal doses of IronQ (75, 150, and 225 µmol Fe/kg BW) over a 14-day period. Pharmacokinetic studies were performed at a dose of 150 µmol Fe/kg BW, with blood iron content analyzed using ICP-OES. For in vivo biodistribution, SD rats were administered an intravenous dose of IronQ (225 µmol Fe/kg BW), followed by MR imaging using a 1.5 T scanner and subsequent tissue-ICP analysis. The longitudinal relaxivity (r1) of IronQ was measured to be 2.17mm⁻¹s⁻¹ in ultrapure water and 3.56mm⁻¹s⁻¹ in whole blood. Acute toxicity studies showed no mortality, morbidity, or significant biochemical changes, with histopathology confirming no irreversible organ damage. Pharmacokinetics revealed peak blood iron content at 1.1hours post-administration and clearance within 24hours. MRI demonstrated enhanced T1 signal intensity, particularly in the liver and kidney. These findings provide valuable insights into the safety, pharmacokinetics, and imaging efficacy of IronQ, highlighting its potential as a robust and biocompatible MRI contrast agent.

In-Silico and In-Vivo Characterization of Anti-Neuro inflammatory potential of Tetrahydrocoptisine by using LPS-Induced model in mice

Neuroinflammation can be directly linked to the imbalance in the Kynurenine-tryptophan Pathway (KP) metabolism. Under inflammatory circumstances, the KP is activated, resulting in a rise in the KP metabolite L-kynurenine (KYN) in the peripheral and central nervous systems (CNS). Increased amounts of KYN in the brain may lead to neurotoxic KYN metabolites, mostly due to breakdown by Kynurenine-3-monooxygenase (KMO). Tetrahydrocoptisine (also known as stylopine) is an alkaloid isolated from Corydalis impatiens. Molecular docking with specific proteins involved in the Neuroinflammation mechanism was studied. LPS-induced neuroinflammation to mice. After 7 days of acclimatization, the animals in groups II, III, and IV were given 5 mg/kg i.p. of the endotoxin LPS. Groups III and IV were subsequently given daily intraperitoneal doses of 18.4 mg/kg and 36.8 mg/kg of our test medication Tetrahydrocoptisine, while group II was used as a disease control. On the 15th day, all groups were assessed neuro-behaviorally. On the 16th day, the mice were slaughtered for histopathology, lipid peroxidation, and nitrite studies. The neurobehavioural assessment involving elevated plus-maze, sucrose preference test, line crossing, and actophotometer revealed that the test drug is capable of decreasing LPS-induced anxiety, depression, and anhedonia at both low and high doses respectively. The histopathological analysis indicated that the neurodegeneration is attenuated at high doses of Tetrahydrocoptisine. A test drug demonstrated potency in inhibiting Kynurenine monooxygenase (KMO) expression in the brain, leading to reduced levels of nitric oxide and lipid peroxidation compared to a control group.

Dimethyl Fumarate Improves Detrusor Contractility in Streptozotocin-Induced Diabetic Rats

Aim: The present study aimed to investigate potential effect of dimethyl fumarate (DMF) on bladder dysfunction in streptozotocin (STZ)-induced diabetic rats. Material and Methods: Adult male Sprague Dawley rats were given a single intraperitoneal dose of streptozotocin (60 mg/kg) to induce diabetes. After eight weeks, diabetic and nondiabetic rats were orally treated with DMF (25 or 100 mg/kg/day) or vehicle for four weeks orally. At 12 week after diabetes induction, in vitro organ bath studies were performed on detrusor strips of each rat and the contractile responses to KCl and carbachol (CCh) of the strips were evaluated. Results: The maximal KCl (80 mM)- and CCh-induced contractile responses of detrusor strips significantly (p

Histopathological and Immunohistochemical Evaluation of Methotrexate-Induced Gonadal Damage in Rats: Role of SCF, mTOR, and SIRT-1.

Methotrexate (MTX) is a highly effective chemotherapy for cancer. This drug has a gonadotoxic effect, mainly in the testes and ovaries. Our study used histopathological and immunohistochemical methods to assess the potential damage to testicular and ovarian tissue caused by MTX use. Twenty-four Wistar albino rats, both male and female, were used in our study. Four sets of rats; control male, MTX male, control female, and MTX female were created. The male and female MTX-treated groups received a single intraperitoneal dose of 20 mg/kg MTX. The testes and ovaries of rats sacrificed under general anesthesia were extracted and histopathologically analyzed. In addition, the immunoreactivity intensities of stem cell factor (SCF), mechanistic target of rapamycin (mTOR), and SIRT-1 in both tissues were measured by immunohistochemistry. Johnsen's testicular biopsy score in the testicular seminiferous tubules was significantly lower in the MTX group than in the control group (p<0.001). The ovary showed substantial follicular degeneration (p<0.05), vascular congestion (p<0.01), and fibrosis (p<0.001). MTX reduced SCF immunoreactivity density in the testis and ovary (p<0.05). Furthermore, MTX reduced mTOR, a marker of autophagy, in the testis (p<0.05) and ovary (p<0.001) compared with the control. SIRT-1 intensity increased dramatically in the testis (p<0.001) and ovary (p<0.01) in the injured group, unlike the mTOR marker. Our investigation revealed that the gonads incurred significant damage as a result of MTX. One vital option for reducing or eliminating this damage to the ovaries and testicles is the use of anti-oxidant-rich substances.

Involvement of necroptosıs and apoptosıs ın protectıve effects of cyclosporın a on ischemıa-reperfusıon injury in rat kıdney.

We aimed to investigate the protective effects of low dose cyclosporin A (CsA) on ischemia-reperfusion (IR) injury in rat the kidney and on the apoptotic and necroptotic mechanisms involved. 1. Control group (received a single intraperitoneal (i.p.) dose of 1ml sterile saline 15min before the surgical procedure), 2. IR group (was subjected to 30min of bilateral kidney ischemia followed by 90min of reperfusion; and received a single i.p. dose of 1ml sterile saline 15min before the IR procedure, 3. IR + CsA group (received a single i.p. dose of 3mg/kg CsA 15min before the IR procedure. Renal functions (renal perfusion pressures, and serum urea-creatinine levels), kidney histological scores, MDA levels, and TNF-α, caspase-3, RIP1, RIP3, MLKL, CaMKII and CypD protein expressions were also measured. Renal perfusion pressures (PP), serum urea and creatinine levels, renal tissue MDA levels, and the protein expression levels of TNF-α, caspase-3, RIP1, RIP3, MLKL, CAMKII and CypD were significantly increased in the IR group compared to the control group (p < 0.05), Additionally, there were significant decreases in all the parameters in the IR + CsA group compared to those in the IR group (p < 0.05). Furthermore, histopathological analyses revealed significantly higher kidney injury scores in the IR group compared to the control group, and low dose CsA treatment improved the injury. A single low dose of CsA injection 15min before IR, demonstrated a protective effect on bilateral renal IR injury and a reduction in apoptotic and necroptopic markers which is resulted in improvement of renal functions.

Alogliptin attenuates STZ-induced diabetic nephropathy in rats through the modulation of autophagy, apoptosis, and inflammation pathways: Targeting NF-κB and AMPK/mTOR pathway.

Diabetic nephropathy (DN) is a type of microvascular complication that arises from diabetes mellitus and leads to further health issues. Most importantly, the prevalence of DN is steadily rising in developed countries. This research explored the therapeutic benefits of alogliptin, a dipeptidyl peptidase IV (DPP-4) inhibitor, on streptozotocin (STZ)-induced DN and its underlying mechanisms in rats. Ten rats were allocated to group 1, served as the normal group; and received saline. To develop diabetes, thirty rats were administered a single intraperitoneal dose of STZ (45mg/kg). STZ-induced diabetic rats were randomly assigned to three groups: group 2 diabetic control; was given saline, groups 3 and 4 received alogliptin (10mg/kg) and (20mg/kg), respectively. The treatment began 8weeks after diabetes onset and continued for four weeks. Histopathological alterations in the kidney were detected. Serum was collected to measure blood glucose levels (BGL), renal function, and lactate dehydrogenase (LDH). Tissue samples were collected to detect changes in oxidative stress (OS), inflammation, 5' adenosine monophosphate-activated protein kinase (AMPK), and the mammalian target of Rapamycin (mTOR) signaling pathways in addition to apoptotic and autophagy changes. Alogliptin reduced STZ-induced histological changes in the kidney as well as OS, and inflammation. Alogliptin also ameliorated the AMPK/mTOR signaling pathways, enhanced autophagy, and reduced apoptosis. These results demonstrate that alogliptin ameliorates inflammation and OS and consequently modulates the AMPK/mTOR axis along with targeting autophagy and apoptosis, leading to the alleviation of DN.

Intestinal failure-associated liver disease model: a reduced phytosterol intravenous lipid emulsion prevents liver injury.

Long-term parenteral nutrition in children often results in intestinal failure-associated liver disease (IFALD). Phytosterols are plant steroids in vegetable oil-based intravenous lipid emulsions (ILEs) that are associated with IFALD. We investigated whether a phytosterol-depleted soybean oil ILE, compared to standard soybean oil ILE, prevented hepatotoxicity in a murine IFALD model. Eight-week-old male C57BL/6 J mice were provided a fat-free high carbohydrate liquid dietfor 19 days. Mice were intravenously administered ILEs as the sole fat source: Intralipid® (commercially available soybean oil ILE), Omegaven® (commercially available fish oil ILE), a low phytosterol soybean oil ILE (L-SOLE) or a high phytosterol soybean oil ILE (H-SOLE) with matched alpha tocopherol content. On days 6, 12, and 18 mice were administered escalating intraperitoneal doses of lipopolysaccharide. Compared to chow controls, mice that received Intralipid® demonstrated elevated plasma biomarkers of liver injury and histologic liver disease (hepatosteatosis, histologic inflammation, F4/80 staining). L-SOLE prevented both biochemical and histologic liver injury. Administration of H-SOLE also prevented biochemical liver injury, but not steatosis. The combination of phytosterol removal and alpha tocopherol supplementation may reduce the toxicity associated with parenteral use of soybean oil-based ILE. Low phytosterol soybean oil may be a valuable component in safer next generation ILEs. Half of children receiving long-term parenteral nutrition develop intestinal failure-associated liver disease (IFALD). Standard intravenous lipid emulsions (ILEs) in parenteral nutrition are vegetable oil based and high in phytosterols (plant steroids); no low phytosterol vegetable oil-based ILE is available. Phytosterols in ILEs are associated with IFALD. In this study, a new phytosterol-depleted soybean oil was utilized in a laboratory-generated ILE. Use of the phytosterol-depleted soybean oil ILE prevented liver injury in a murine model of IFALD. Phytosterol-depleted soybean oil may be utilized as a component of less toxic next-generation ILEs.

Comparative Study of Non-invasive Mouse Models of Pancreatitis.

Although a relevant animal model is essential for studying human diseases, one has yet to be established for mouse pancreatitis. Early non-invasive models of mouse pancreatitis have serious limitations. In this study, we compared the efficiency, consistency, and reproducibility of inducing pancreatitis in 3 non-invasive mouse models of pancreatitis in Wistar albino mice: (1) L-arginine-induced model (2 intraperitoneal injections of 4 g/kg body weight of L-arginine spaced 1 h apart), (2) caerulein-induced model (6 intraperitoneal injections of 50 µg/kg body weight of caerulein at hourly intervals), and (3) caerulein + LPS (lipopolysaccharide)-induced model (6 intraperitoneal doses of 50 µg/kg body weight of caerulein at hourly intervals, along with an LPS [10 mg/kg body weight] injection immediately after the last caerulein injection). Our findings showed that the L-arginine-induced model was inconsistent. The levels of the pancreatic enzymes, amylase and lipase, were higher in the caerulein and caerulein + LPS groups. Histological examination showed tissue destruction in the induced groups, with varying degrees of fibrosis in the caerulein + LPS group. The caerulein + LPS model was the most reliable model in Wistar albino mice. Our findings may be useful in helping investigators choose the most appropriate animal model for pancreatitis research.

Carmaphycin B-Based Proteasome Inhibitors to Treat Human African Trypanosomiasis: Structure-Activity Relationship and In Vivo Efficacy.

The proteasome is essential for eukaryotic cell proteostasis, and inhibitors of the 20S proteasome are progressing preclinically and clinically as antiparasitics. We screenedTrypanosoma brucei, the causative agent of human and animal African trypanosomiasis, in vitro with a set of 27 carmaphycin B analogs, irreversible epoxyketone inhibitors that were originally developed to inhibit thePlasmodium falciparum20S (Pf20S). The structure-activity relationship was distinct from that of the human c20S antitarget by the acceptance of d-amino acids at the P3 position of the peptidyl backbone to yield compounds with greatly decreased toxicity to human cells. For the three most selective compounds, binding to the Tb20S β5 catalytic subunit was confirmed by competition with a fluorescent activity-based probe. For one compound, J-80, with its P3 d-configuration, the differential binding to the parasite's β5 subunit was supported by both covalent and noncovalent docking analysis. Further, J-80 was equipotent against both Trypanosoma brucei gambiense and Trypanosoma brucei rhodesiense in vitro. In a mouse model of Stage 1 T. brucei infection, a single intraperitoneal (i.p.) dose of 40 mg/kg J-80 halted the growth of the parasite, and when given at 50 mg/kg i.p. twice daily for 5 days, parasitemia was decreased to below the detectable limit, with parasite recrudescence 48 h after the last dose. The in vivo proof of principle demonstrated by a potent, selective, and irreversible inhibitor of Tb20S reveals an alternative path to the development of kinetoplastid proteasome inhibitors that differs from the current focus on allosteric reversible inhibitors.