Histopathological Scores Research Articles

The Effect of Curcumin Nanoparticles on Paracetamol-Induced Liver Injury in Male Wistar Rats.

Curcumin is a naturally occurring compound that has antioxidant properties, acts as a hepatoprotective, and lowers lipid peroxidation. However, curcumin's low solubility and bioavailability are its primary drawbacks and prevent its use as a therapeutic agent. In this study, curcumin nanoparticles will be created using the ultrasonic-assisted extraction method, and their effectiveness against paracetamol-induced changes in ALT, AST, SOD, MDA, and TNF-α will be compared to that of pure curcumin. This study aimed to determine the hepatoprotective effect of curcumin nanoparticles in paracetamol-induced rats as a model for liver injury. Thirty-six male Wistar rats, aged 6 to 8 weeks, with a minimum weight of 120 grams, were used in an experimental laboratory investigation with a post-test-only group design. Rats in each group received 100 mg/kgBW pure curcumin, 100 mg/kgBW curcumin nanoparticles, and 50 mg/kgBW curcumin nanoparticles for 7 days before paracetamol induction. On day 8, 300 mg/kgBW of paracetamol was intraperitoneally injected to cause liver damage. One of the groups received NAC as an antidote 10 hours after paracetamol induction. Detection of ALT and AST using a Chemistry Analyzer. ELISA approach for the detection of SOD, MDA, and TNF-α. The Roenigk score was calculated by two examiners after the liver histopathology preparations were stained using the Hematoxylin-Eosin method. Post hoc analyses were performed after the One Way Annova and Kruskal Wallis tests to examine the data. According to PSA results, the smallest formula that formed curcumin nanoparticles (10.2 nm) was 8 g of curcumin formula mixed with a mixture of Tween 20 4.5 ml, Kolliphor EL 1.5 ml, Propylene Glycol 1.5 ml, and Capryol 90 1 ml for 21 minutes using an ultrasonic process. MDA and TNF-α levels, as well as the liver's histological Roenigk score, were significantly lower in the 100 mg/kgBB pure curcumin group (C100) when compared to the model group (model). The levels of AST, MDA, TNF-α, and the liver histopathology score were significantly lower in the 100 mg/kgBB (NC100) and 50 mg/kgBB (NC50) curcumin nanoparticle groups compared to the model group (model) and pure curcumin group (C100) (p< 0.05). Curcumin nanoparticles showed better hepatoprotective ability than pure curcumin.

Effects of rosmarinic acid on cyclophosphamide-induced nephrotoxicity in rats

Purpose: Cyclophosphamide (CP) is an antineoplastic agent. It is used in the treatment of many types of cancer. Rosmarinic acid (RA) exhibits remarkable biological activities such as anti-inflammatory, antitumor, antibacterial, and antimicrobial effects. This study aimed to evaluate the effect of rosmarinic acid against CP-induced nephrotoxicity.&#x0D; Materials and Methods: Eighteen male Sprague Dawley rats were randomly divided into 3 equal groups; Sham group (n=6): 0.9% saline solution/8 days/oral gavage + 0.9% saline solution/8th day/intraperitoneal, CP group (n=6): 0.9% saline solution/8 days/oral gavage + 200 mg/kg/8th day/intraperitoneal CP, and CP+RA group (n=6): 100 mg/kg/8 days/oral gavage RA + 200 mg/kg/8th day/intraperitoneal CP was applied. Hematoxylin and Eosin, Periodic Acid-Schiff, and Masson’s Trichrome staining were performed on the collected tissues &#x0D; Results: Histopathological evaluation revealed tubular atrophy, glomerular damage, vascular congestion, vacuolization, and interstitial inflammation in the CP group. Histopathological scores were significantly lower in the CP+RA group compared to the CP group. Intertubular fibrosis was observed in the CP group compared to the Sham group. Fibrosis decreased with rosmarinic acid. PAS-stained sections from the CP group showed tubular epithelial vacuolization, brush border, and basal membrane disruption. These findings decreased with rosmarinic acid. The increased blood urea nitrogen level in the CP group was lower in the CP+RA group, while the decreased SOD level in the CP group was higher in the CP+RA group. &#x0D; Conclusion: RA has protective effects against CP causing tubular atrophy, glomerular damage, vascular congestion, vacuolization, and interstitial inflammation in the kidney

Dragon fruit (Hylocereus undatus (Haw.) Britt.) powders attenuate ulcerative disease symptoms via dextran sulfate sodium-induced mouse model

Dragon fruit is cactus based fruit that has loads of health benefits such as lowering risks of a blood sugar spike, aids in digestion etc. Its nutritional content is rich in vitamin C, vitamin E, magnesium, iron etc. Its functions on the physiological regulation are well known. Inflammatory bowel diseases (IBD) are multifactorial chronic intestinal disorders. Currently, mesalamine etc. and therapeutic strategies were suggested for IBD therapy. However, the etiology of IBD remains unclear which is an ongoing challenge and side effects of therapeutic drugs must be also considered. Thus, the aim of this study was evaluated the efficacy and therapeutic strategies investigations on the attenuated IBD symptoms via administrating three doses of dragon fruit powders in the 2% dextran sulfate sodium (DSS)-induced IBD mouse model. The female C57BL/6 mice were divided respectively the normal control group (n = 10), the negative control group (n = 10), three dose groups (n = 10 per group) of dragon fruit powders (250 mg/kg BW, 500 mg/kg BW, and 1,000 mg/kg BW). Exception of the normal control group, other groups were administrated with 2% DSS for 5 days. Later, the normal drinking water was provide to C57BL/6 mice until the end of the experiment. At the end of the experiment, the body weight (BW), the stool appearance/status, the macroscopic and microscopic colonic injuries, and myeloperoxidase (MPO) activity were monitored, measured and scored. The results were showed that BW of C57BL/6 mice in the negative control group, three dragon fruit powder groups was gradually reduced during the IBD period induced by 2% DSS, and BW of C57BL/6 mice gradually increased when the 2% DSS in drinking water was replaced with the normal drinking water. When the experiment was carried out to the 3rd to 4th week, BW of the negative control group was significantly lower than that of the normal control group. The stool appearance/status was presented that stool score in the negative control group was significantly higher than that in the normal control group (p &lt; 0.001). The stool score in the high-dose dragon fruit powder group was significantly lower than that in the negative control group (p &lt; 0.001). The macroscopic colons of C57BL/6 mice were performed at the end of the experiment. (1) Gut weight: It can be seen that gut weight in the normal control group is lowest and the gut weight in the negative control group is highest between all groups. The gut weight in the negative control group was higher than that in the normal control group, medium-dose dragon fruit powder group, and high-dose dragon fruit powder group were seen. (2) Gut length: It can be seen that gut length in the normal control group is longest. The gut length in the normal control group is significantly longer than that in the other groups. Exception of the normal control group, other groups were not significant difference compared to each other. (3) Gut weight-to-gut length ratio: It can be seen that gut weight-to-gut length ratio in the normal control group is lowest and the gut weight-to-gut length ratio in the negative control group is highest between all groups. The gut weight-to-gut length ratio in the negative control group was significantly higher than that in the normal control group (p &lt; 0.01). The gut weight-to-gut length ratio in the high-dose dragon fruit powder group was significantly lower than that in the negative control group. The microscopic colons of C57BL/6 mice was performed at the end of the experiment. The pathological analysis items were divided into ulcer area ratio, mucosal ulcer depth, inflammatory cell infiltration, and submucosal edema. Total histopathologic scores in the 2% DSS-induced group was also significantly higher than that of the normal control group. Finally, the evaluating MPO activity was performed by using MPO activity assay kit. It can be seen that MPO activity was significant higher in the negative control group than that in the normal control group. MPO activity in the three dragon fruit powder groups were significant lower than that in the negative control group. Taken all results together, the consumption of medium-dose (500 mg/kg BW) and high-dose (1,000 mg/kg BW) dragon fruit powders has a positive improvement effect on relieving various symptoms caused by IBD via a successful 2% DSS-induced IBD mouse model.

Experimental Investigation of Lung Toxicity after Radiation Therapy Combined with Immune Checkpoint Inhibitors

The combination of radiation therapy (RT) with immune checkpoint inhibitors (ICIs) has resulted in prolonged survival in patients with locally advanced lung cancer (NSCLC). However, RT and ICIs carry the risk of inflammatory and irreversible lung damage and might have synergistic effects on these adverse events. Importantly, only little is known about the risk of enhanced side effects of the lung and which factors modulate such toxicity. In addition, there is a need for sensitive diagnostics for the early detection of lung injury after combined radioimmunotherapy. Recently, experimental x-ray dark-field radiography was able to detect radiation-induced lung injury earlier than conventional radiography in mice. The right thorax of mice was irradiated with unfractionated RT (1x15 Gy or 1x30 Gy) or fractionated RT (5x9 Gy). In addition, indicated experimental groups received ICIs (a-PD1, a-CTLA4 or combination) for one month. After four months, the mice were analyzed using different methods to quantify lung damage: lung coefficient as surrogate marker for lung fibrosis, histopathological staining of fibrosis, conventional x-ray, and dark-field radiography. We found enhanced signs of lung fibrosis (lung coefficient and histopathological score) after unfractionated RT in combination with ICIs. Surprisingly, combination of ICIs with RT resulted in opposite effects. Specifically, concomitant combination of ICIs with fractionated RT resulted in reduced lung coefficients and lower histopathological signs of lung fibrosis in mice treated in this way. Importantly, in vivo x-ray dark-field radiographs showed the same trend as the ex vivo assessment of the lungs. To the best of our knowledge, this is the first experimental study to find that the combination of RT with ICIs has an impact on the risk of pulmonary fibrosis. Strikingly, ICIs (a-PD1 + a-CTLA4) combined with unfractionated RT resulted in increased lung damage, while concomitant use of ICIs (a-PD1 + a-CTLA4) with fractionated RT resulted in reduced lung toxicity. In the field of radioimmunotherapy, such studies are of great value for the interpretation of clinical studies and of importance for the design of further clinical trials.

High expression of P-selectin induces neutrophil extracellular traps via the PSGL-1/Syk/Ca2+/PAD4 pathway to exacerbate acute pancreatitis.

Excessive neutrophil extracellular traps (NETs) is involved in the progression of acute pancreatitis (AP) but the mechanisms controlling NETs formation in AP are not fully understood. Therefore, our study sought to investigate the mechanism of the highly expressed P-selectin stimulating the formation of NETs in AP. NETs formation was detected by flow cytometry, immunofluorescence staining, and cf-DNA and MPO-DNA complexes were measured as biomarkers of NETs formation. Neutrophils treated with P-selectin and pharmacological inhibitors were examined by western blot, immunofluorescence staining and flow cytometry. Mouse model of AP was established by caerulein and the effect of inhibiting P-selectin by PSI-697 on the level of NETs and PAD4 in pancreatic tissue was observed. The severity of AP was evaluated by histopathological score and the detection of serum amylase and lipase. Patients with AP had elevated levels of NETs and P-selectin compared with healthy volunteers. Stimulation of P-selectin up-regulated the expression of PSGL-1, increased the phosphorylation of Syk, mediated intracellular calcium signal and led to the activation and expression of PAD4, which modulated NETs formation in neutrophils. Pretreament with PSI-697 blunted NETs formation and PAD4 expression in the pancreatic tissue, and ameliorated the severity of AP in mice. Taken together, these results suggest that P-selectin induces NETs through PSGL-1 and its downstream Syk/Ca2+/PAD4 signaling pathway, and that targeting this pathway might be a promising strategy for the treatment of AP.

Therapeutic efficacy of β-sitosterol treatment on Trypanosoma congolense infection, anemia development, and trans-sialidase (TconTS1) gene expression.

African animal trypanosomiasis hinders sustainable livestock productivity in sub-Saharan Africa. About 17 million infected cattle are treated with trypanocides annually but most of the drugs are associated with drawbacks, necessitating the search for a promising chemotherapeutic agent. In this study, the effects of β-sitosterol on Trypanosoma congolense infection were investigated along with its effect on the trans-sialidase gene expressions. Oral treatment with β-sitosterol at 15 and 30 mg/kg body weight (BW) for 14 days significantly (p < 0.05) reduced parasitemia and ameliorated the parasite-induced anemia. Also, the parasite-induced increase in serum urea level and renal histopathological damage scores in addition to renal hypertrophy was significantly (p < 0.05) reverted following treatment with 30 mg/kg BW β-sitosterol. The compound also significantly (p < 0.05) down-regulated the expression of TconTS1 but not TconTS2, TconTS3, and TconTS4. Correlation analysis between free serum sialic acid with the TconTS1 and TconTS2 gene variants revealed negative correlations in the β-sitosterol-treated groups although they were non-significant (p > 0.05) in the group treated with 15 mg/kg BW β-sitosterol. Similarly, a non-significant negative (p > 0.05) correlation between the biomolecule and the TconTS3 and TconTS4 gene variants was observed in the β-sitosterol-treated groups while positive correlations were observed in the infected untreated control group. The observed effect of β-sitosterol on T. congolense infection could make the compound a possible template for the design of novel trypanocides.

The Effects of Probiotic Lactobacillus rhamnosus GG on Fecal Flora and Serum Markers of Renal Injury in Mice with Chronic Kidney Disease.

In this study, we analyzed intestinal flora in an experimental mouse model of chronic kidney disease (CKD) and investigated whether oral supplementation with probiotic Lactobacillus rhamnosus GG could slow the decline in renal function and inflammatory status of mice with CKD. We surgically induced chronic kidney disease in C57BL/6J male mice aged 8-9 weeks. We used dual-stage 5/6 nephrectomy for this, while the mock group underwent a mock procedure. The experimental (CKD mice) and mock group were administered a daily dose of 10 × 109 colony forming unit (CFU) of probiotic L. rhamnosus GG or 2 g of maltodextrin as a placebo by oral gavage, respectively, for 5 weeks. At the end of the experiment, the fecal samples of the mice were collected and prepared for intestinal microbial diversity analysis. We examined the serum chemistry and renal histology of the mice. Important serum and blood biomarkers were associated with the development of CKD, including increased serum concentrations of creatine, cystatin C, blood urea nitrogen (BUN), and a protein-interleukin-6 (denoted as IL-6), whereas decreased serum albumin concentration was also observed in the mice with CKD. The intestinal flora of the mice with CKD significantly declined in terms of diversity, richness, and homogeneity. The consumption of L. rhamnosus GG probiotic via oral gavage significantly decreased the serum concentration level present in creatinine and blood urea nitrogen. However, it increased albumin in the group with CKD. After probiotic treatment, serum IL-6 levels dropped considerably, and the kidney histopathology score in mice with CKD who were given L. rhamnosus GG improved. Moreover, supplementation with the probiotic significantly improved floral richness and lineage diversity in the mice with CKD.Conclusions: In this study, we found that probiotics significantly attenuated renal failure development, reduced serum levels of proinflammatory cytokine IL-6, and increased the abundance and lineage diversity of intestinal flora in mice with chronic kidney disease.

Effect of Kangfuxiaomi suppository on pelvic inflammatory disease in rats

Pelvic inflammatory disease (PID) is commonly encountered in gynecological practice. Kangfuxiaomi suppository, made from the compound extract of Periplaneta Americana, is a Traditional Chinese Medicine remedy widely used for the treatment of gynecological disorders. This study aimed to preliminarily explore the therapeutic effect of Kangfuxiaomi suppository in a rat model of PID established by chemical injury and pathogen infection. The key parameters assessed were vulvar inflammation score, vaginal + uterine organ index, and serum levels of interleukin (IL)− 8; tumor necrosis factor (TNF)-α; C-reactive protein (CRP); superoxide dismutase (SOD); and malondialdehyde (MDA). In addition, levels of IL-6, cyclooxygenase (COX)− 2, and IL-2 in cervical tissues as well as that of IL-1β and prostaglandin E-2 (PGE2) in uterine tissues were measured. The expression levels of nuclear factor-kappa B (NF-κB) p65 and Toll-like receptor 4 (TLR4) in uterine tissues were detected by immunohistochemical method. After Kangfuxiaomi suppository treatment, the vulva inflammation score and histopathological score of PID rats showed a tendency to decrease. Serum IL-8, TNF-α, CRP, and MDA levels were reduced, while SOD levels were significantly increased. Levels of IL-6, IL-2, and COX-2 in cervical tissues were somewhat decreased, and PGE2 and IL-1β levels in uterine tissue were significantly decreased. Moreover, the levels of NF-κB p65 and TLR4 protein expression were also decreased. These findings demonstrated the therapeutic effect of Kangfuxiaomi suppository in PID rats. The underlying mechanism may involve enhanced antioxidant capacity and decreased secretion of proinflammatory factors via the NF-κB/TLR4 signaling pathway.

Co-administration of curcumin and polyamines in high-fat diet induced obese rats: Assessment of changes in serum polyamine levels and some tissue parameters

Obesity is a non-communicable chronic disease that continues to increase around the world. Recently, it has been shown that curcumin positively affects lipid, energy metabolism, and body weight change. Moreover, polyamines are aliphatic polycations, which can be found in all mammalian cells and foods and have been shown to prevent obesity through many different mechanisms. However, whether the co-administration of curcumin and polyamines has synergistic effects has yet to be clarified. Our study aimed to examine the effects of curcumin and polyamines on obesity and to assess the changes in serum polyamine levels and tissue parameters. 28 Sprague-Dawley male rats were fed a high-fat diet for 10 weeks to develop obesity, and then they were randomly divided into 4 groups as the control group (CONT), curcumin group (CUR), polyamine group (POL), curcumin and polyamine group (CUR+POL) and supplements were administered for 6 weeks. As a result, the lowest feed consumption in rats was recorded in the CUR+POL group, and the group with the lowest weight after supplements was the POL group, then the CUR+POL, CONT, and CUR groups, respectively. N-acetyl putrescine and GABA levels increased significantly after obesity development. The total histopathological score in fat, liver, and kidney tissues increased significantly in the CONT group. In the CUR+POL group, damage to the tissues was in the direction of recovery compared to the other groups, and the expression of NF-κB was significantly low. These results suggest that combined curcumin and polyamines may have protective effects.

Streamlining quantitative joint-wide medial femoro-tibial histopathological scoring of mouse post-traumatic knee osteoarthritis models

Histological scoring remains the gold-standard for quantifying post-traumatic osteoarthritis (ptOA) in animal models, allowing concurrent evaluation of numerous joint tissues. Available systems require scoring multiple sections/joints making analysis laborious and expensive. We investigated if a single section allowed equivalent quantitation of pathology in different joint tissues and disease stages, in three ptOA models. Male 10-12-week-old C57BL/6 mice underwent surgical medial-meniscal-destabilization, anterior-cruciate-ligament (ACL) transection, non-invasive-ACL-rupture, or served as sham-surgical, non-invasive-ACL-strain, or naïve/non-operated controls. Mice (n = 12/group) were harvested 1-, 4-, 8-, and 16-weekpost-intervention. Serial sagittal toluidine-blue/fast-green stained sections of the medial-femoro-tibial joint (n = 7/joint, 84 µm apart) underwent blinded scoring of 40 histology-outcomes. We evaluated agreement between single-slide versus entire slide-set maximum or median scores (weighted-kappa), and sensitivity/specificity of single-slide versus median/maximum to detect OA pathology. A single optimal mid-sagittal section showed excellent agreement with median (weighted-kappa 0.960) and maximum (weighted-kappa 0.926) scores. Agreement for individual histology-outcomes was high with only 19/240 median and 15/240 maximum scores having a weighted-kappa ≤0.4, the majority of these (16/19 and 11/15) in control groups. Statistically-significant histology-outcome differences between ptOA models and their controls detected with the entire slide-set were reliably reproduced using a single slide (sensitivity >93.15%, specificity >93.10%). The majority of false-negatives with single-slide scoring were meniscal and subchondral bone histology-outcomes (89%) and occurred in weeks 1-4 post-injury (84%). A single mid-sagittal slide reduced the time needed to score diverse histopathological changes by 87% without compromising the sensitivity or specificity of the analysis, across a variety of ptOA models and time-points.

Effect of chronic low-dose treatment with chitooligosaccharides on microbial dysbiosis and inflammation associated chronic ulcerative colitis in Balb/c mice

The study aimed to investigate the potential of low dose chitooligosaccharide (COS) in ameliorating dextran sodium sulfate (DSS) induced chronic colitis by regulating microbial dysbiosis and pro-inflammatory responses. Chronic colitis was induced in BALB/c mice by DSS (4% w/v, 3 cycles of 5 days) administration. The mice were divided into four groups: vehicle, DSS, DSS + mesalamine and DSS+COS. COS and mesalamine were administered orally, daily once, from day 1 to day 30 at a dose of 20 mg/kg and 50 mg/kg respectively. The disease activity index (DAI), colon length, histopathological score, microbial composition, and pro-inflammatory cytokine expression were evaluated. COS (20 mg/kg, COSLow) administration reduced the disease activity index, and colon shortening, caused by DSS significantly. Furthermore, COSLow restored the altered microbiome in the gut and inhibited the elevated pro-inflammatory cytokines (IL-1 and IL-6) in the colon against DSS-induced chronic colitis in mice. Moreover, COSLow treatment improved the probiotic microflora thereby restoring the gut homeostasis. In conclusion, this is the first study where microbial dysbiosis and pro-inflammatory responses were modulated by chronic COSLow treatment against DSS-induced chronic colitis in Balb/c mice. Therefore, COS supplementation at a relatively low dose could be efficacious for chronic inflammatory bowel disease.

Fraxin in Combination with Dexamethasone Attenuates LPS-Induced Liver and Heart Injury and Their Anticytokine Activity in Mice.

Cytokine storm syndrome (CSS) is a major cause of morbidity and mortality in people suffering from hyperinflammatory status, which diverse etiological factors, including pathogens, therapeutic interventions, malignancies, and autoimmune disorders, can instigate. Since there is limited research on the antioxidant properties of fraxin and no studies have investigated its potential as an anticytokine storm agent, it is important to note that most studies have primarily focused on proinflammatory cytokines such as IL-1β, IL-6, and TNFα during cytokine storm. However, little research discusses the role of chemokines, particularly IL-8, during cytokine storms. Therefore, further investigation is warranted into the role of fraxin as an anticytokine storm agent and the involvement of IL-8 in cytokine storms. The present study examines the preventive efficacy of fraxin and the combination of fraxin and dexamethasone (FD) in mitigating lipopolysaccharide-induced systemic inflammation in mice caused by Escherichia coli, 055: B5. Five groups of ten mice were randomly assigned: LPS only group (5 mg/kg, intraperitoneally i.p.), control (normal saline N.S. 1 ml/kg, i.p.), concentrations were selected based on previous literature, fraxin (120 mg/kg, i.p.), dexamethasone (5 mg/kg, i.p.), fraxin + dexamethasone (FD) (60 mg/kg + 2.5 mg/kg, i.p.), administered one hour before LPS injection (5 mg/kg,i.p.), animals were euthanized next day, and interleukin-8 (IL-8) was quantified in serum using an enzyme-linked immunosorbent assay. The liver and heart tissues underwent histopathological analysis to assess morphological changes. For data analysis using ANOVA and Tukey post hoc tests, the Kruskal-Wallis and Mann-Whitney U tests were employed to analyze the histological results. A significant decline in IL-8 levels was recorded in the treatment groups almost to the same degree (p < 0.001), and the percentage of inhibition of IL-8 for fraxin, dexamethasone, and FD was 93%.92.4%, and 93%, respectively, compared to the LPS-only group. Histopathological scores were significantly reduced in liver and heart tissue (P < 0.05). All interventions used in this study significantly reduced interleukin-8 (IL-8) levels and reduced LPS-induced liver and cardiac damage. The combination (FD) did not result in an evident superiority of either agent. More research is required to identify the possible usefulness of these agents in treating hyperinflammatory diseases, such as cytokine storms, in future clinical practice.

ALDH1A Inhibition Suppresses Colitis and Alters α4β7 Integrin Expression on Activated T Cells in Mdr1a-/- Mice.

There are limited pharmacological treatment options for inflammatory bowel disease (IBD), and some of these options are expensive and administered by injection or infusion. Thus, new cheaper and easier (oral) treatment options are needed. ALDH1A enzymes produce retinoic acid that can affect intestinal diseases such as IBD by regulating immune cells in the gut. We previously demonstrated that an orally deliverable ALDH1A inhibitor, WIN 18,466, can suppress colitis in an acute mouse model of IBD. Here, we tested the efficacy of ALDH1A inhibition in a chronic mouse model of IBD. Mdr1a-/- mice were treated with a diet containing WIN 18,446 starting 1 week prior to inducing colitis by H. bilis inoculation. Treatment was continued until the study end point and colitis was monitored based on clinical symptoms and confirmed by histological analysis. Immune cell phenotypes in colon-draining lymph nodes (cMLN) were analyzed. WIN 18,446 treatment reduced clinical symptoms and improved histopathologic colitis scores. This was associated with decreased expression of the gut homing integrin, α4β7, on T cells in cMLN; increased expression of CD103, a protein associated with tissue-resident memory T cells; and changes in dendritic cells, plasmacytoid dendritic cells and B cells in inhibitor-treated mice. ALDH1A inhibition broadly influences immune cells during colitis and is a potential new target for IBD treatment. Future studies will be needed to determine the efficacy of ALDH1A inhibition on active colitis and to evaluate its relative efficacy in comparison to approved drugs.

Effects of adriamycin on cell differentiation and proliferation in rat testis

ABSTRACT Although adriamycin (ADR) is used to treat many cancers, it can be toxic to healthy organs including the testis. We investigated the effects of ADR on pluripotency in rat testis. Testicular damage was induced by either cumulative or single dose single dose administration of ADR in Wistar albino rats. Rats were divided randomly into three groups: untreated control, cumulative dose ADR group (2 mg/kg ADR every three days for 30 days) and single dose ADR group (15 mg/kg, single dose ADR). Testicular damage was evaluated and seminiferous tubule diameters were measured using light microscopy. Expression levels of Oct4, Sox2, Klf4, c-Myc, Utf1 and Dazl were assessed by immunohistochemistry and real time PCR. Serum testosterone levels were measured using ELISA assay. Histopathologic scores were lower and mean seminiferous tubule diameters were less compared to the ADR groups. Oct4, Sox2, Klf4 and Utf1 expressions were decreased significantly in spermatogenic cells of both cumulative and single dose ADR groups compared to the control group. We found that c-Myc expression in spermatogenic and Leydig cells were increased significantly in both ADR groups compared to the control group. Dazl expression was decreased in the cumulative adriamycin group compared to the control group, but increased in the single dose ADR group compared to both the control and cumulative ADR groups. Serum testosterone levels were decreased in both ADR groups compared to the control group. Our findings suggest that ADR is detrimental to regulation and maintenance of pluripotency in rat testis.

Benefits of accurate and guided endoureterotomy versus ureteral balloon dilatation in the management of ureteral strictures-comparative animal study.

Endoscopic treatment of ureteral strictures provides a minimally invasive approach with a shorter hospital stay and less postoperative pain. There are different therapeutic options, the dilatation balloon and endoureterotomy with holmium yttrium-aluminum-garnet (Ho:YAG) laser are the most used. To assess histological changes after endoureterotomy in the ureteral stricture treatment comparing Ho:YAG laser endoureterotomy versus balloon dilatation endoureterotomy. The subjects used were a total of 48 female pigs. The initial assessment consisted of an endoscopic, nephrosonographic, and contrast fluoroscopic evaluation of the urinary tract. Subsequently, a model of ureteral stricture was performed. Three weeks later, the ureteral stricture was diagnosed and treated. Then animals were randomly assigned to two groups (group A, Balloon dilatation endoureterotomy and group B, Holmium laser retrograde endoureterotomy) in which a double-pigtail ureteral stent was placed for 3 weeks. Follow-up assessments were performed at 3-6 weeks. The final follow-up was completed at 5 months and included the pathological study. In terms of therapeutic effectiveness, the overall success was 81.2%. The success rate was 91.7% in group B and 70.8%in group A without statistical significance. No evidence of vesicoureteral reflux nor urinary tract anomalies were observed. Histological assessment showed statistical significance in overall score, lamina propria fibrosis and serosal alterations in group A with higher histological changes. The overall histopathological score after ureteral stricture treatment in an animal model showed better remodeling of incised ureteral wall healing after Ho:YAG laser endoureterotomy. Laser endoureterotomy tends to have higher success rate compared to balloon dilatation.

Aerosolized, Gas-Phase, Intravitreal Methotrexate Reduces Proliferative Vitreoretinopathy in a Randomized Trial in a Porcine Model.

To determine the effectiveness of aerosol-delivered methotrexate (AD-MTx) in a large-animal (porcine) model of proliferative vitreoretinopathy (PVR). Prospective, randomized, interventional, double-masked, controlled, large-animal study with predetermined clinical and histopathologic outcome criteria. Half of the pigs were randomly assigned to receive an identical volume of aerosol-delivered normal saline (AD-NS) using identical delivery systems and treatment intervals. Proliferative vitreoretinopathy was surgically induced in 16 pigs (8 males and 8 females), randomly assigned to receive 2 doses (group A) or 3 doses (group B) of either AD-MTx (1.6 mg/0.4 ml) or normal saline (AD-NS). Group A pigs were euthanized at week 2 (n= 8), and group B pigs were euthanized at week 3 (n= 8). Masked clinical PVR scores (0-6) by a vitreoretinal surgeon and histopathology PVR scores (0-8) by a masked ophthalmic pathologist were used to determine outcomes. The mean, combined clinical and histopathology scores (both anterior and posterior) were used to determine the overall treatment effect between the groups. The mean masked score (± standard deviation) when all grading end points (clinical+ histopathology) were combined was a mean of 8.0 ± 2.3 in the AD-MTx group versus a higher 9.9 ± 2.0 in the AD-NS control group (P= 0.05). The clinical score was 3.88 ± 1.2 in the AD-MTx group versus 4.63 ± 1.6 in the AD-NS group (P= 0.16). The histopathology score for anterior PVR was 2.5 ± 0.8 in the AD-MTx group versus 2.5 ± 0.5 in the AD-NS group (P= 0.50), and the posterior PVR was 1.63 ± 1.6 in the AD-MTx group versus 2.75 ± 1.3 in the AD-NS group (P= 0.07). When the frequency of methotrexate dosing in group A (2 doses) was compared with that in group B (3 doses), the mean score was 8.75 versus 9.13 (P= 0.38), respectively, suggesting an insignificant difference. After surgical induction of PVR in an aggressive, high-risk, large-animal model, AD-MTx reduced posterior PVR formation compared with AD-NS. Additional dosing at week 3 did not improve the outcomes. No difference in anterior PVR formation was noted with intervention. This novel drug delivery system has implications for PVR reduction and warrants further investigation. Proprietary or commercial disclosure may be found after the references.

Anti-rheumatic and Memory-enhancing Effect of Memantine on Complete Freund’s Adjuvant-induced Rheumatoid Arthritis Compared to Methotrexate in Wistar Rats

Objectives To evaluate the anti-inflammatory and memory-enhancing effects of memantine (MEM) in complete Freund’s adjuvant (CFA)-induced rheumatoid arthritis (RA) in Wistar rats. The effect of MEM on cognition in CFA-induced RA was also evaluated. Materials and Methods The rats were randomly assigned to one of five groups: sham control (SC), disease control (DC)—CFA 0.1 mg, positive control (PC)—methotrexate (MTX) 1 mg/kg, low dose MEM—20 mg/kg, and high dose MEM (HD-M)—50 mg/kg. All groups received a subcutaneous injection of CFA in the left paw on day 0, except for the SC group, which received a distilled water (DW) injection. The study drugs, MEM and MTX, were administered orally from days 15 to 28. The SC group received DW orally from days 15 to 28. Variables for physical (paw volume, paw surface temperature, and ankle joint diameter) and behavior (open field test, grip strength test, and water corn test) were performed on days 1, 0, 7, 14, 21, and 28. On day 28, blood was collected for serum inflammatory markers, tumor necrosis factor-alpha (TNF-α) and interleukin (IL)-6. Ex vivo radiological changes in the ankle joint were recorded using a digitalized X-ray machine. Histopathology of the ankle joint was performed. Results When compared to the DC group, the HD-M group showed a significant improvement in all physical and behavioral characteristics ( p &lt; 0.001). When compared to the DC group, TNF-α, IL-6 levels, and histopathological scores were significantly lower in the HD-M group ( p &lt; 0.01). Conclusion MEM reduces inflammation and improves memory in Wistar rats with CFA-induced RA. MEM can be used as an anti-inflammatory drug in elderly patients at risk of dementia.

Fasudil and SR1001 synergistically protect against sepsis-associated pancreatic injury by inhibiting RhoA/ROCK pathway and Th17/IL-17 response

Sepsis is defined as a dysregulated host response to infection that can result in organ dysfunction and high mortality, which needs more effective treatment urgently. Pancreas is one of the most vulnerable organs in sepsis, resulting in sepsis-associated pancreatic injury, which is a fatal complication of sepsis. The aim of this study was to investigate the effect of combination of fasudil and SR1001 on sepsis-associated pancreatic injury and to explore the underlying mechanisms. The model of sepsis-associated pancreatic injury was induced by cecal ligation and puncture. Pancreatic injury was evaluated by HE staining, histopathological scores and amylase activity. The frequency of Th17 cells was analyzed by flow cytometry. Serum IL-17 level was determined by ELISA. Protein levels of RORγt, p-STAT3, GEF-H1, RhoA and ROCK1 were determined by Western blot. The apoptosis of pancreatic cells was examined by TUNEL analysis and Hoechst33342/PI staining. Compared to the sham group, the model group showed significant pathological injury including edema, hyperemia, vacuolization and necrosis. After treatment with fasudil, model mice showed an obvious reduction of Th17 cells and IL-17. SR1001 significantly reduced the expressions of GEF-H1, RhoA and ROCK1 in the model mice. The combination treatment with fasudil and SR1001 significantly inhibited the differentiation of Th17 cells, expressions of IL-17, GEF-H1, RhoA and ROCK1, which were more effective than each mono-treatment. In addition, our data revealed a remarkable decrease of apoptosis in pancreatic acinar cells culturing with fasudil or SR1001, which was further inhibited by their combination culture. Lipopolysaccharide remarkably upregulated the differentiation of Th17 cells in vitro, which could be significantly downregulated by fasudil or SR1001, and further downregulated by their combination treatment. Taken together, the combination of fasudil with SR1001 has a synergistic effect on protecting against sepsis-associated pancreatic injury in C57BL/6 mice.

Nano-selenium impacts on growth performance, digestive enzymes, antioxidant, immune resistance and histopathological scores of Nile tilapia, Oreochromis niloticus against Aspergillus flavus infection

The present study examined the impacts of varying amounts of selenium nanoparticles (Se-NPs) as a natural antioxidant and metabolic regulator on growth performance, antioxidant capacity, digestive enzymes and immune resistance of Oreochromis niloticus challenged to Aspergillus flavus infection. For 60 days, fish were fed the experimental diet of 30% crude protein up to apparent satiation three times a day. Se-NPs were added to the control diet at different levels of 0.0 (control), 0.5 (T1), 1.0 (T2) and 1.5 (T3) mg/kg diet to fed monosex Nile tilapia fingerlings with average initial weight (4.5 ± 0.5 g). Fish were randomly distributed in 12 tanks 100 L at a density of 20 fish per tank after 2-week acclimation to represent four treatments in triplicates. The results indicated that the growth indices, feed efficiency and survival rate were significantly enhanced (P < 0.05) by incorporating Se-NPs up to 1 mg/kg diet. Furthermore, the haemato-biochemical parameters, digestive enzymes activity and antioxidant capacity of the fish were significantly improved (P < 0.05) at T2. When the fish were challenged with Aspergillus flavus, a decreasing mortality rate was observed, which clearly shows that selenium nanoparticles boosted the fish’s immune response at T2. The intestinal morphology as villus length, villus width, muscular fibres layer thickness and absorption intestinal zone after challenged with A. flavus infection were considerably enhanced (P < 0.05) by incorporating all Se-NP levels. Histopathological score significantly improved (P < 0.05) for the hepatopancreatic, intestinal, gills and muscle tissues at T2. The present study concludes that selenium nanoparticles up to 1.0 mg/kg diet can be efficiently used in tilapia feed to help boost fish production, immune system response and histopathological parameters.

Targeting Autophagy, Apoptosis, and SIRT1/Nrf2 Axis with Topiramate Underlies Its Neuroprotective Effect against Cadmium-Evoked Cognitive Deficits in Rats.

Cadmium is an environmental toxicant that instigates cognitive deficits with excessive glutamate excitatory neuroactivity in the brain. Topiramate, a glutamate receptor antagonist, has displayed favorable neuroprotection against epilepsy, cerebral ischemia, and Huntington's disease; however, its effect on cadmium neurotoxicity remains to be investigated. In this study, topiramate was tested for its potential to combat the cognitive deficits induced by cadmium in rats with an emphasis on hippocampal oxidative insult, apoptosis, and autophagy. After topiramate intake (50 mg/kg/day; p.o.) for 8 weeks, behavioral disturbances and molecular changes in the hippocampal area were explored. Herein, Morris water maze, Y-maze, and novel object recognition test revealed that topiramate rescued cadmium-induced memory/learning deficits. Moreover, topiramate significantly lowered hippocampal histopathological damage scores. Mechanistically, topiramate significantly replenished hippocampal GLP-1 and dampened Aβ42 and p-tau neurotoxic cues. Notably, it significantly diminished hippocampal glutamate content and enhanced acetylcholine and GABA neurotransmitters. The behavioral recovery was prompted by hippocampal suppression of the pro-oxidant events with notable activation of SIRT1/Nrf2/HO-1 axis. Moreover, topiramate inactivated GSK-3β and dampened the hippocampal apoptotic changes. In tandem, stimulation of hippocampal pro-autophagy events, including Beclin 1 upregulation, was triggered by topiramate that also activated AMPK/mTOR pathway. Together, the pro-autophagic, antioxidant, and anti-apoptotic features of topiramate contributed to its neuroprotective properties in rats intoxicated with cadmium. Therefore, it may be useful to mitigate cadmium-induced cognitive deficits.