Intestinal Histopathological Changes Research Articles

Autoinducer-2 May Be a New Biomarker for Monitoring Neonatal Necrotizing Enterocolitis.

Autoinducer-2 (AI-2) has a widely accepted role in bacterial intra- and interspecies communication. Little is known about the relationships between AI-2 and NEC. This study found that AI-2 levels in patients and in a NEC mouse model were detected using the Vibrio harveyi BB170 assay system. Bacterial communities of the newborns' stool microbiota (NEC acute group, NEC recovery group, control group, and antibiotics-free group) and of the NEC mouse model (NEC group and control group) were detected by high-throughput sequencing. Intestinal histopathological changes were observed after HE staining. The AI-2 level in the NEC acute group (44.75 [40.17~65.52]) was significantly lower than that in the control group, NEC recovery group and antibiotics-free group. The overall microbiota compositions of each group at the phylum level were not significantly different. The proportions of Enterococcus, Clostridium_sensu_stricto_1, Peptoclostridium, and Veillonella had significant differences among the 4 groups at the genus level. In animal experiments, the AI-2 level in feces of NEC mice (56.89 ± 11.87) was significantly lower than that in the feces of control group mice (102.70 ± 22.97). The microbiota compositions of NEC and control group mice at the phylum level were not significantly different. At the genus level, Klebsiella, Clostridium_sensu_stricto_1, and Peptoclostridium abundances in the NEC group increased significantly compared with those in the control group (P < 0.05). In addition, Lactobacillus, Pasteurella, and Parabacteroides abundances in the NEC group decreased significantly compared with those in the normal control group (P < 0.05), while Lactobacillus, Pasteurella, and Parabacteroides abundances had the opposite trend. The AI-2 concentration decreased significantly in the acute phase of NEC and increased gradually in the convalescent phase. We conclude that the concentration of AI-2 was correlated with intestinal flora disorder and different stages of disease. AI-2 may be a new biomarker for the diagnosis and monitoring of NEC.Trial Registry: ClinicalTrials.gov; ChiCTR-ROC-17013746; URL: www.clinicaltrials.gov

Encephalitozoon intestinalis: A new target for auranofin in a mice model.

Despite the fact that many approaches have been developed over years to find efficient and well-tolerated therapeutic regimens for microsporidiosis, the effectiveness of current drugs remains doubtful, and effective drugs against specific targets are still scarce. The present study is the first that was designed to evaluate the potency of auranofin, an anti-rheumatoid FDA approved drug, against intestinal Encephalitozoon intestinalis. Evaluation of the drug was achieved through counting of fecal and intestinal spores, studying the intestinal histopathological changes, measuring of intestinal hydrogen peroxide level, and post therapy follow-up of mice for 2 weeks for detection of relapse. Results showed that auranofin has promising anti-microsporidia potential. It showed a promising efficacy in mice experimentally infected with E. intestinalis. It has revealed an obvious reduction in fecal spore shedding and intestinal tissue spore load, amelioration of intestinal tissue pathological changes, and improvement of the local inflammatory infiltration without significant changes in hydrogen peroxide level. Interestingly, auranofin prevented the relapse of infection. Thus, considering the results of the present work, auranofin could be considered a therapeutic alternative for the gold standard drug 'albendazole' against the intestinal E. intestinalis infection especially in relapsing cases.

Dietary administration of gelatinised polyhydroxybutyrate to Penaeus vannamei improved growth performance and enhanced immune response against Vibrio parahaemolyticus

The effect of dietary supplementate, gelatinized polyhydroxy butyrate on the growth performance, and prophylactic disease control in Penaeus vannamei was evaluated. Healthy juvenile shrimp were collected from farms, acclimatized in a laboratory aquarium and fed gelatinized polyhydroxybutyrate (PHB) diet for 60 days. Infectivity experiments were conducted with Vibrio parahaemolyticus SF14 which was isolated from diseased shrimp samples. The treated, control and challenged shrimp samples were analyzed for intestinal function, immune enzymes, and histopathological changes. The PHB treated shrimp showed significant weight gain (7.81 ± 0.184 g) compared to the commercial diet fed control shrimp (5.914 ± 0.070 g) at 5% significant level. The activity of immune enzymes such as prophenol oxidase and superoxide dismutase activity along with the total antioxidant activity significantly elevated in the PHB diet fed shrimp. The digestive enzyme levels were found to be higher in the PHB treated shrimp indicates the PHB modulates digestive function in shrimp. The V. parahaemolyticus SF14 challenge resulted in 100% mortality in the control shrimp whereas the PHB diet fed shrimp were completely protected with 100% survival against V. parahaemolyticus infection. A histopathological examination revealed that shrimp fed with the commercial diet and challenged with V. parahaemolyticus displayed necrotic muscles and vacuolation in the muscle fibers along with hemocyte infiltration in the gills. In contrast, the shrimp fed PHB diet and challenged with V. parahaemolyticus showed no pathological changes in the muscle fibers and gills. Cumulatively, this study demonstrated that dietary supplementation of 1% gelatinized PHB effectively protects shrimp against infective pathogens and improves their immune function and growth rate.

Low dose Epigallocatechin Gallate Alleviates Experimental Colitis by Subduing Inflammatory Cells and Cytokines, and Improving Intestinal Permeability.

Background: In this study, we investigate the impact of epigallocatechin gallate (EGCG), the most abundant and potent catechin in green tea, on a mouse model of inflammatory bowel disease (IBD) and the underlying mechanisms of action. Methods: C57BL/6J mice were subjected to dextran sulfate sodium (DSS)-induced IBD-like disease and then randomly divided into three groups: Model group (MD), low-dose EGCG group (LE, 20 mg/kg/d), and high-dose EGCG group (HE, 50 mg/kg/d). DSS-induced clinical and macroscopic changes were monitored daily. Intestinal permeability was assessed by FITC-Dextran assay. Results: Both high- and low-dose EGCG treatment alleviated clinical manifestations including body weight loss and disease activity index (DAI) of DSS-induced colitis. The DAI score was significantly improved after two days of EGCG treatment. At the end of the study, the macroscopic severity score (MSS) of HE and LE treatment groups were 2.4 ± 1.2, and 2.2 ± 1.0, respectively, significantly lower than that of the controls (5.0 ± 2.1). EGCG treatment also prevented colon shortening, and improved intestinal permeability and histopathological changes. In addition, EGCG treatment attenuated colon inflammation by suppressing colonic levels of pro-inflammatory cytokines IL-6, MCP-1, and TNF-alpha, and inhibited CD3+ T cell and CD68+ macrophage infiltration. Conclusion: EGCG is effective in inflammatory colitis because it reduces cellular and molecular inflammation, and reduces intestinal permeability.

Production and functional insights into the potential regulation of three isoforms of grass carp p40 subunit in inflammation

The p40 subunit is known as a component of Interleukin (IL)-12 and IL-23. In mammals, p40 can be secreted as a monomer or homodimer and acts independently to mediate cellular responses. Recently, three p40 paralogues were isolated and identified from grass carp and other fish species, but whether they exist independently as well as their functional consequences and significance remain unclear. In the present study, using grass carp as the model, we for the first time demonstrated the existence of natural fish p40a, p40b and p40c (gcp40a, gcp40b and gcp40c) mainly as a monomer in culture supernatant of head kidney leukocytes (HKLs). Particularly, their excessive secretion induced by various immune stimuli suggests possible involvement of free p40s in fish immune responses. To define their functions, recombinant grass carp p40a/b/c (rgcp40a, rgcp40b and rgcp40c) were prepared by Pichia pastoris expression system, and they possessed the activities to enhance the secretion of pro-inflammatory cytokines including Il-1β and tumor necrosis factor-α (Tnf-α) in grass carp HKLs. These pro-inflammatory properties of p40 isoforms prompted us to investigate their roles during the inflammatory process. In line with this, in vivo study revealed the pathogenic effect of rgcp40a on intestinal inflammation, whereas gcp40a polyclonal antibodies remarkably ameliorated Aeromonas hydrophila-induced intestinal histopathological changes. Taken together, our results uncover the biological significance of free p40s in teleost, and provide new clue for targeting fish intestinal inflammation.

Fisetin and telmisartan each alone or in low-dose combination alleviate OVA-induced food allergy in mice.

Food allergy (FA) is a worldwide health problem, affecting nearly 10% of all populations, with no prophylactic options or regulatory treatment available until now. Fisetin, a biologically active flavonoid, and telmisartan, the highly selective competitive AT1 receptor antagonist, recently exhibited potent anti-inflammatory and immunomodulatory activities. In the present study, we have evaluated the possible anti-inflammatory and immunomodulatory activities of fisetin and telmisartan each alone or in low-dose combination in a mouse model of FA. For induction of FA, eight-week-old BALB/c mice, sensitized by two ip injection of 50 μg ovalbumin (OVA) and 1 mg alum at day 0 and 7. Then, each mouse challenged with 10 mg OVA at days 14, 16, 18, and 21. On the 28th day, the fifth challenge carried out by oral administration of 50 mg OVA. Either fisetin (1 or 3 mg/kg/d), telmisartan (1 or 3 mg/kg/d) or a combination of fisetin 1 mg/kg/d and telmisartan 1 mg/kg/d received orally from the 13th day till 28th day. In challenge days, the treatments received one-hour before the challenge. Our data showed that fisetin and telmisartan each alone or in low-dose combination attenuated the anaphylactic manifestation, decreased blood eosinophilic count, serum OVA-specific IgE, and IL-4 levels, the intestinal total and degranulated mast cells count, and CD4+ immunohistochemical expression. Furthermore, they enhanced the serum IFN-γ level and abrogated the intestinal histopathological changes induced by OVA in mice. Either fisetin, telmisartan or their low-dose combination could be promising in the management of FA.

THERAPEUTIC EFFICACY OF CHITOSAN NANOPARTICLES AND ALBENDAZOLE IN INTESTINAL MURINE TRICHINELLOSIS

Most of the drugs used for the treatment of trichinellosis showed a limited bioavailability and a high degree of resistance. Therefore, there is an urgent need to develop new agents to improve the bioavailability of these drugs. This work assessed the use Chitosan (CH) nanoparticles singly or combined with albendazole (ABZ) for the treatment of experimental trichinellosis. Fifty male albino mice were used. They were divided into six experimental groups. Chitosannanoparticles were used orally at a dose of 100mg/kg/day starting from 3rd dpi for three successive days for the treatment of the intestinal phase of infection either alone or loaded with full dose or half dose of ABZ. Results showed significant reduction in all treated groups with the highest reduction rate of adult 99.1%, improvement of the intestinal histopathological changes and a significant decrease in iNOS expression of the intestinal tissue were observed ingroups treated by CH loaded with full dose of ABZ compared to control group

THERAPEUTIC EFFICACY OF CHITOSAN NANOPARTICLES AND ALBENDAZOLE IN INTESTINAL MURINE TRICHINELLOSIS

Most of the drugs used for the treatment of trichinellosis showed a limited bioavailability and a high degree of resistance. Therefore, there is an urgent need to develop new agents to improve the bioavailability of these drugs. This work assessed the use Chitosan (CH) nanoparticles singly or combined with albendazole (ABZ) for the treatment of experimental trichinellosis. Fifty male albino mice were used. They were divided into six experimental groups. Chitosannanoparticles were used orally at a dose of 100mg/kg/day starting from 3rd dpi for three successive days for the treatment of the intestinal phase of infection either alone or loaded with full dose or half dose of ABZ. Results showed significant reduction in all treated groups with the highest reduction rate of adult 99.1%, improvement of the intestinal histopathological changes and a significant decrease in iNOS expression of the intestinal tissue were observed ingroups treated by CH loaded with full dose of ABZ compared to control group

Atorvastatin: In-Vivo Synergy with Metronidazole as Anti-Blastocystis Therapy.

Blastocystis is an enteric Straminopile in tropical, subtropical and developing countries. Metronidazole has been a chemotheraputic for blastocystosis. Failures in its regimens were reported and necessitate new studies searching for alternative therapeutic agents. Aim of current study is to investigate potential effects of Atorvastatin (AVA) compared to the conventional chemotherapeutic MTZ in experimentally Blastocystis-infected mice. Anti-Blastocystis efficacy of AVA was evaluated parasitologically, histopathologically and by transmission electron microscopy using MTZ (10 mg/kg) as a control. Therapeutic efficacy of AVA was apparently dose-dependent. Regimens of AVA (20 and 40 mg/kg) proved effective against Blastocystis infections with high reduction in Blastocystis shedding (93.4–97.9%) compared to MTZ (79.3%). The highest reductions (98.1% and 99.4%) were recorded in groups of combination treatments AVA 20–40 mg/kg and MTZ 10 mg/kg. Blastocystis was nearly eradicated by the 20th day post infection. Genotype analysis revealed that genotype I was most susceptible, genotype III was less. Histopathologic and ultrastructural studies revealed apoptotic changes in Blastocystis and significant improvement of intestinal histopathological changes more remarkable in combinational therapy groups. Thus, the present study offers AVA as a potential candidate for Blastocystis therapy combined with MTZ.

A new scope for orlistat: Effect of approved anti-obesity drug against experimental microsporidiosis.

As the current therapies for intestinal microsporidiosis are either inconsistent in their efficacies or hampered by several adverse effects, alternative antimicrosporidial agents are being sought. The present study is the first that was designed to evaluate the potency of orlistat, an approved anti-obesity drug, against intestinal microsporidiosis caused by both Enterocytozoon bieneusi and Encephalitozoon intestinalis. Results were assessed through studying fecal and intestinal spore load, intestinal histopathological changes, viability, and infectivity of spores from treated animals. Results showed that orlistat has promising antimicrosporidia potential, with better results in E. intestinalis than E. bieneusi. The animals that received orlistat showed statistically significant decrease in the fecal and intestinal spore load, when compared to the corresponding control infected nontreated mice. The results were insignificant compared to fumagillin and albendazole. Light microscopic examination of stained intestinal sections revealed amelioration of the pathological changes and decreased inflammatory cells detected in the control infected nontreated mice. Spores encountered from stool of orlistat-treated E. bieneusi and E. intestinalis mice showed low viability and significant reduction of infectivity versus their control. Thus, considering the results of the present work, orlistat proved its effectiveness against the intestinal microsporidial infection.

Supplementation of Vitamin E Protects Chickens from Newcastle Disease Virus-Mediated Exacerbation of Intestinal Oxidative Stress and Tissue Damage

Background/Aims: Newcastle disease virus (NDV) causes a highly devastating and contagious disease in poultry, which is mainly attributed to extensive tissue damages in the digestive, respiratory and nervous systems. However, nature and dynamics of NDV-induced oxidative stresses in the intestine of chickens remain elusive. Methods: In this study, we examined the magnitude of intestinal oxidative stress and histopathological changes caused by the virulent NDV infection, and explored the protective roles of vitamin E (vit. E) in ameliorating these pathological changes. For these purposes, chickens were divided into four groups namely i) non supplemented and non-challenged (negative control, CON); ii) no supplementation of vit. E but challenged with ZJ1 (positive control, NS+CHA); iii) vit. E supplementation at the dose of 50 IU/day/Kg body weight and ZJ1 challenge (VE50+CHA); and 4) vit. E supplementation at the dose of 100 IU/day/Kg body weight and ZJ1 challenge (VE100+CHA). In all groups, we analyzed concentrations of glutathione (GSH), malondialdehyde (MDA), nitric oxide (NO), total antioxidant capacity (T-AOC), and activity of glutathione S-transferase (GST), superoxide dismutase (SOD), catalase (CAT) using biochemical methods. The virus loads were determined by quantitative RT-PCR and antibody titers by hemagglutination inhibition assays. We also examined the histopathological changes in the duodenal and jejunal mucosa at 3 and 5-day post infection (dpi) with NDV. Results: A significant elevation in the NO level was observed in NDV challenged chickens compared to the CON chickens at 2 dpi. The MDA contents were significantly increased whereas GSH was significantly decreased in NDV-challenged chickens compared to control. Furthermore, activities of GST, CAT, SOD, as well as the TOAC were markedly decreased in challenged chickens in comparison with control. Virus copy numbers were higher in NDV infected NS+CHA group compared to other groups. Severe histopathological changes including inflammation, degeneration and broken villi were observed in the intestine of NDV challenged chickens. However, all these malfunctions of antioxidant system and pathological changes in the intestine were partially or completely reversed by the vit. E supplementation. Conclusions: Our results suggest that NDV infection causes oxidative stress and histopathological changes in the duodenum and jejunum of chickens, which can be partially or fully ameliorated by supplementation of vit. E. Additionally, these findings suggest that oxidative stress contributes to the intestinal damages in NDV infected chickens. These findings will help to understand the pathogenesis of NDV and further investigation of therapeutic agents for control of Newcastle disease.

Protective effect of salvianolic acid B against intestinal ischemia reperfusion-induced injury in a rat model

Purpose: To evaluate the gastro-protective efficacy of salvianolic acid B (SAB) against intestinal ischemic-reperfusion injury (IIRI) in a rat model.Methods: Forty-eight healthy male rats were randomly choosen and divided into 4 groups of 12 rats each. Control group rats underwent laparotomy without occlusion; IIRI group rats underwent laparotomy with occlusion for 60 min, followed by 24 h of reperfusion; SAB + IIRI group received 7 days of pretreatment with 40 mg/kg of SAB + IIRI; while the fourth group received only SAB. The antioxidant, inflammatory markers, intestinal permeability marker, as well as intestinal histopathological changes were assessed.Results: The activities of antioxidants including reduced glutathione (GSH), catalase (CAT) and superoxide dismutase (SOD) were significantly ameliorated (p &lt; 0.01) in SAB-supplemented group (SAB + IIRI). The concentration of inflammatory markers, including interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-α) and nuclear factor p65 (NF-p65) as well as small intestinal permeability marker (FITC-Dextran), were significantly reduced (p &lt; 0.01) following administration of SAB for 7 days. In addition, pretreatment with SAB reverted intestinal (ileum) histopathological changes to almost normal architecture with significant reduction in Chiu score.Conclusion: The results of this study demonstrate that SAB may protect the intestine by attenuating oxidative stress and inflammatory response and hence, may be potentially for treating IIRI.Keywords: Salvianolic acid B, Intestinal Ischemia-reperfusion, Antioxidants, Inflammation, Intestinal permeability

17β-Estradiol prevents mesenteric injury induced by occlusion of the proximal descending aorta in male rats

ObjectiveIn surgical aortic repair or cardiac surgery with aorta occlusion, the occurrence of mesenteric ischemia and bowel injury has been associated with higher short-term mortality. The vascular protection of estrogens has been investigated and is mainly mediated by increasing the availability of nitric oxide (NO). Therefore, this study investigated the role of 17β-estradiol on visceral ischemia-reperfusion (I/R) injury after descending aorta occlusion in male rats. MethodsMesenteric ischemia was induced in male Wistar rats by placing a 2F Fogarty arterial embolectomy catheter (Edwards Lifesciences, Irvine, Calif) in the descending aorta, which remained occluded for 15 minutes, followed by reperfusion for up to 2 hours. Rats were divided into four groups: (1) rats that underwent surgical manipulation only (sham, n = 22); (2) rats that underwent I/R injury (n = 22); (3) rats treated with intravenous 17β-estradiol (280 μg/kg) 30 minutes before I/R (n = 22); (4) or at the beginning of reperfusion (n = 22). Intestinal histopathologic changes were evaluated by histomorphometry. Mesenteric microcirculatory alterations were assessed by laser Doppler flowmetry and intravital microscopy technique. Protein expression of intercellular adhesion molecule-1, P-selectin, endothelial NO synthase (eNOS), and endothelin-1 was evaluated by immunohistochemistry; in addition, eNOS and endothelin-1 gene expressions were quantified by real-time polymerase chain reaction. Serum cytokines were measured by enzyme-linked immunosorbent assay. ResultsRelative to the sham group, the I/R group exhibited a highly pronounced loss of intestine mucosal thickness, a reduction in mesenteric blood flow (P = .0203), increased migrated leukocytes (P < .05), and high mortality rate (35%). Treatment with 17β-estradiol before aorta occlusion preserved intestine mucosal thickness (P = .0437) and mesenteric blood flow (P = .0251), reduced the number of migrated leukocytes (P < .05), and prevented any fatal occurrence. Furthermore, 17β-estradiol downregulated the expression of intercellular adhesion molecule-1 (P = .0001) and P-selectin (P < .0001) on the endothelium and increased the protein expression of eNOS (P < .0001). The gene expressions of eNOS and endothelin-1 did not differ between the groups. ConclusionsThe prophylactic treatment with 17β-estradiol showed better overall repercussions and was able to prevent any fatal occurrence, increase eNOS expression, thus preserving mesenteric perfusion and intestinal integrity, and reduce inflammation.

Postconditioning: “Toll-erating” mesenteric ischemia-reperfusion injury?

Postconditioning may prove to be a suitable method to decrease ischemia-reperfusion injury of intestine after mesenteric arterial occlusion. Toll-like-receptor-4 is involved in the pathophysiology of organ damage after ischemia-reperfusion; therefore, the aim of our study was to investigate the effect of postconditioning on the mucosal expression of toll-like-receptor-4. Male Wistar rats (n=10/group) underwent 60minutes of superior mesenteric artery occlusion followed by 6hours of reperfusion in 3 groups: sham-operated, ischemia-reperfusion, and a postconditioned group. Postconditioning was performed by 6 alternating cycles of 10seconds of reperfusion/reocclusion. Blood and tissue samples were collected at the end of reperfusion. Intestinal histopathologic changes and immunohistochemical expression of mucosal caspase-3, antioxidant status, and protein levels of high-mobility group box-1 and toll-like-receptor-4 were assessed. Immunofluorescent labeling and confocal microscopic analysis of toll-like-receptor-4 were performed. Mucosal and serum levels of interleukin-6 and tumor necrosis factor-α protein were measured. Histologic alterations in the postconditioned group were associated with decreased caspase-3 positivity, less toll-like-receptor-4 mRNA, and less protein expression of high-mobility group box-1 and toll-like-receptor-4 in the intestinal villi compared with the ischemia-reperfusion group. Furthermore, a significantly improved antioxidant state of the intestinal mucosa and less mucosal and serum protein levels of interleukin-6 and tumor necrosis factor-α were detected in the postconditioned group. Small intestinal ischemia-reperfusion injury in male Wistar rats caused by the occlusion of the superior mesenteric artery was ameliorated by the use of postconditioning, showing a more favorable inflammatory response, which may be attributed to the decreased mucosal expression of toll-like-receptor-4.

The protective effect of Aplysin on ethanol‐induced liver damage via modulation of liver Kupffer cell function and altered intestinal flora in rats

Aplysin, a member of the bromine‐sesquiterpene compounds family, was mainly extracted from red alga Laurencia tristicha. This study aimed to explore the effects of Aplysin on rat liver, intestine, Kupffer cells and intestinal flora in rats exposed to alcohol. Rats were randomly divided into three groups: normal control group, ethanol‐model group, and Aplysin‐treated group. Except control group, rats were exposed to ethanol at dose of 6 g/kg/d for first four weeks, then 8 g/kg/d for last four weeks. One hour after that, Aplysin‐treated group was given oral administration of Aplysin at 150 mg/kg/bw on a daily basis. After 8 weeks, the rats were sacrificed and liver and intestine were collected for histological and biochemical assessments, plasma was collected for endotoxin level measurement with commercial kit, primary rat Kupffer cell cultures were established for measuring expression of CD14, TLR4 and NF‐kB using RT‐PCR and production of TNF‐α and IL‐1β in liver Kupffer cells using ELISA. Fecal samples were collected for quantitative analysis of intestinal floras. We found that Aplysin reduced the alcohol‐induced hepatic and intestinal histopathological changes and plasma endotoxin levels, and also suppressed the elevation of biomarkers for liver damage (AST, ALT, ALP, CHE and GGT). In primary Kupffer cell cultures, asplysin significantly decreased expression of CD14, TLR4, and NF‐kB as well as production of TNF‐α and IL‐1β. In addition, Aplysin significantly affected the changes in intestinal floras, including Escherichia coli, Enterococcus faecalis, Lactobacillus and Bifidobacterium. In conclusion, Aplysin exerted potent liver‐protective effect, which is associated with modulation of Kupffer cells function and altered intestinal flora.Support or Funding InformationThis work was supported by Grants from the National Nature Science Foundation of China (No.81573137) and Dietary Nutrition Research and Education Foundation of Danone Nutrition Center (No. DIC2014‐03)

Carbonic anhydrase enzyme as a potential therapeutic target for experimental trichinellosis.

Trichinellosis is a globally distributed helminthic infection. There is a considerable interest in developing new anti-helminthic drugs affecting all the developmental stages of Trichinella. Acetazolamide (carbonic anhydrase (CA) inhibitor) involves a novel mechanism of action by inhibiting such an essential enzyme for parasite metabolism. This work aimed to study the effect of acetazolamide against different stages of T. spiralis in experimental animals. Mice were divided into three groups: group I: infected and treated with acetazolamide on day 2 post infection (P.I.), group II: infected and treated with acetazolamide on day 12 P.I., and group III: infected non-treated. From each group, small intestine and muscles were removed for histopathological and immunohistochemical studies. Also, total adult and muscle larval count were estimated. We found that acetazolamide was effective in reduction of both adult and muscle larval counts. When given early, the effect was more pronounced on the adults (62.7%). However, the efficacy of the drug against muscle larvae was increased when given late (63%). Improvement of the intestinal histopathological changes was observed in all the treated groups. Degeneration of encysted larvae with minimal pathologic changes of infected skeletal muscle was observed in the treated groups. Expression of matrix metalloproteinase-9 showed a statistically significant decrease in the intestinal and muscle tissues in all treated groups as compared to the control group. In conclusion, the present study revealed that acetazolamide, carbonic anhydrase inhibitor, could be a promising drug against both adults and larvae of T. spiralis.

Global hypoxia-ischemia induced inflammation and structural changes in the preterm ovine gut which were not ameliorated by mesenchymal stem cell treatment.

Perinatal asphyxia, a condition of impaired gas exchange during birth, leads to fetal hypoxia-ischemia (HI) and is associated with postnatal adverse outcomes including intestinal dysmotility and necrotizing enterocolitis (NEC). Evidence from adult animal models of transient, locally-induced intestinal HI has shown that inflammation is essential in HI-induced injury of the gut. Importantly, mesenchymal stem cell (MSC) treatment prevented this HI-induced intestinal damage. We therefore assessed whether fetal global HI induced inflammation, injury and developmental changes in the gut and whether intravenous MSC administration ameliorated these HI-induced adverse intestinal effects. In a preclinical ovine model, fetuses were subjected to umbilical cord occlusion (UCO), with or without MSC treatment, and sacrificed 7 days after UCO. Global HI increased the number of myeloperoxidase positive cells in the mucosa, upregulated mRNA levels of interleukin (IL)-1β and IL-17 in gut tissue and caused T-cell invasion in the intestinal muscle layer. Intestinal inflammation following global HI was associated with increased Ki67+ cells in the muscularis and subsequent muscle hyperplasia. Global HI caused distortion of glial fibrillary acidic protein immunoreactivity in the enteric glial cells and increased synaptophysin and serotonin expression in the myenteric ganglia. Intravenous MSC treatment did not ameliorate these HI-induced adverse intestinal events. Global HI resulted in intestinal inflammation and enteric nervous system abnormalities which are clinically associated with postnatal complications including feeding intolerance, altered gastrointestinal transit and NEC. The intestinal histopathological changes were not prevented by intravenous MSC treatment directly after HI, indicating that alternative treatment regimens for cell-based therapies should be explored.

Effect of dietary supplementation with clay-based binders on biochemical and histopathological changes in organs of turkey fed with aflatoxin-contaminated diets.

This study was carried out to investigate the effect of dietary supplementation with molecular or nano-clay binders on biochemical and histopathological examination of organs of turkeys fed diets contaminated with aflatoxin B1. Two hundred and sixteen unsexed 1-day-old British United Turkeys were randomly allotted to nine diets in a 3×3 factorial arrangement of diets supplemented with no toxin binder, molecular toxin binder (MTB) and nano-clay toxin binder, each contaminated with 0, 60 and 110ppb aflatoxin B1 respectively. There were three replicates per treatment with eight turkeys per replicate. Biochemical analyses, organ weights and histopathological changes of some organs were examined at the end of the study which lasted for 84 days. Turkeys fed diets supplemented with molecular and nano-binders showed higher (p<0.001) total serum protein, reduced (p<0.001) serum uric acid and GGT concentration values when compared with those fed aflatoxin-contaminated diets supplemented with no binder. Turkeys fed aflatoxin-contaminated diets supplemented with no binder had increased (p<0.001) AST and ALT concentration when compared with other treatments. The heaviest (p<0.001) liver and intestinal weight was noticed with turkeys fed diets supplemented with no binder and contaminated with 110ppb aflatoxin B1 . Pathologically, there was no visible morphological alteration noticed in all turkeys fed uncontaminated diets and nano-clay-supplemented group. Hepatic paleness, hepatomegaly and yellowish discolouration of the liver were observed with turkeys fed diets containing no binder but contaminated with 60 and 110ppb aflatoxin B1. Intestinal histopathological changes such as goblet cell hyperplasia, villous atrophy and diffuse lymphocytic enteritis were more prominent in turkeys fed diets containing no toxin binder and MTB. In conclusion, there were improved biochemical parameters and reduced deleterious effects of aflatoxin B1 in turkeys fed diet supplemented with clay binders. However, the improvement was more conspicuous in the nano-clay-supplemented group than molecular clay group.

Sevoflurane Preconditioning Reduces Intestinal Ischemia-Reperfusion Injury: Role of Protein Kinase C and Mitochondrial ATP-Sensitive Potassium Channel

Ischemic preconditioning (IPC) has been considered to be a potential therapy to reduce ischemia-reperfusion injury (IRI) since the 1980s. Our previous study indicated that sevoflurane preconditioning (SPC) also reduced intestinal IRI in rats. However, whether the protective effect of SPC is similar to IPC and the mechanisms of SPC are unclear. Thus, we compared the efficacy of SPC and IPC against intestinal IRI and the role of protein kinase C (PKC) and mitochondrial ATP-sensitive potassium channel (mKATP) in SPC. A rat model of intestinal IRI was used in this study. The superior mesenteric artery (SMA) was clamped for 60 min followed by 120 min of reperfusion. Rats with IPC underwent three cycles of SMA occlusion for 5 min and reperfusion for 5 min before intestinal ischemia. Rats with SPC inhaled sevoflurane at 0.5 minimum alveolar concentration (MAC) for 30 min before the intestinal ischemic insult. Additionally, the PKC inhibitor Chelerythrine (CHE) or mKATP inhibitor 5-Hydroxydecanoic (5-HD) was injected intraperitoneally before sevoflurane inhalation. Both SPC and IPC ameliorated intestinal IRI-induced histopathological changes, decreased Chiu’s scores, reduced terminal deoxyribonucleotide transferase-mediated dUTP nick end labeling (TUNEL) positive cells in the epithelium, and inhibited the expression of malondialdehyde (MDA) and tumor necrosis factor-α (TNF-α). These protective effects of SPC were similar to those of IPC. Pretreatment with PKC or mKATP inhibitor abolished SPC—induced protective effects by increasing Chiu’s scores, down-regulated the expression of Bcl-2 and activated caspase-3. Our results suggest that pretreatment with 0.5 MAC sevoflurane is as effective as IPC against intestinal IRI. The activation of PKC and mKATP may be involved in the protective mechanisms of SPC.

Effects of hydrogen inhalation on serum pro-inflammatory factors and intestinal injury in mice with severe sepsis

To investigate the effects and mechanisms of hydrogen inhalation on serum levels of pro-inflammatory factors and intestinal injury in severe septic mice. 176 male ICR mice were randomly divided into four groups: sham operation group, hydrogen control group (sham+hydrogen inhalation), model group (severe sepsis model) and hydrogen treatment group (severe sepsis model+hydrogen inhalation), with 44 mice in each group. Severe sepsis model was reproduced by cecal ligation and puncture (CLP). 2% hydrogen inhalation was given for 1 hour at 1 hour and 6 hours after sham or CLP operation. Twenty animals in each group were selected and observed for 7-day survival rate. Six animals in each group were selected and sacrificed at 6, 12, 24 and 48 hours after sham or CLP, the concentrations of tumor necrosis factor-α (TNF-α), interleukins (IL-6, IL-10) and high mobility group box 1 (HMGB1) in serum were determined, the intestinal histopathological changes and scores were evaluated by light microscopy, and the activities of myeloperoxidase (MPO) and caspase-3 were determined. The 7-day survival rate of severe sepsis mice was 0; the 7-day survival rate was increased to 60% in hydrogen treatment group, with statistical significance in variables compared with model group (P<0.05). Compared with sham operation group, the serum concentrations of TNF-α, IL-6, IL-10 and HMGB1 were obviously increased, the intestine were heavily injured along with higher histopathological scores, and the intestinal MPO and caspase-3 activities were significantly enhanced at different time points after CLP in model group (all P<0.05). Compared with model group, the serum concentrations of TNF-α, IL-6 and HMGB1 were significantly decreased [ TNF-α (ng/L): 6 hours: 110.34±9.28 vs. 440.55±25.78, 12 hours: 82.29±8.43 vs. 448.36±32.54, 24 hours: 79.68±9.04 vs. 346.42±22.24, 48 hours: 80.79±10.06 vs. 368.94±31.58; IL-6 (ng/L): 12 hours: 58.68±8.55 vs. 158.28±16.73, 24 hours: 46.98±7.58 vs. 146.74±18.02, 48 hours: 38.67±8.22 vs. 136.45±15.45; HMGB1 (μg/L): 6 hours: 15.75±4.32 vs. 55.56±10.04, 12 hours: 32.02±9.33 vs. 89.65±15.65, 24 hours: 35.87±8.54 vs. 86.44±20.33, 48 hours: 23.85±9.83 vs. 98.33±18.88, all P<0.05], the serum concentrations of IL-10 (ng/L) at 24 hours and 48 hours after CLP were obviously increased (24 hours: 135.44±16.43 vs. 79.22±12.03, 48 hours: 110.92±12.54 vs. 74.47±11.18, both P<0.05), the intestinal injury were ameliorated with decreased histopathological scores (12 hours: 1.70±0.06 vs. 3.23±0.44, 24 hours: 2.12±0.31 vs. 4.51±0.58, 48 hours: 2.03±0.42 vs. 4.27±0.58, all P<0.05), and the intestinal MPO and caspase-3 activities were significantly decreased [MPO (U/g): 6 hours: 13.75±4.21 vs. 25.56±5.34, 12 hours: 14.72±4.22 vs. 30.53±6.87, 24 hours: 11.62±3.14 vs. 33.58±7.24, 48 hours: 11.33±4.03 vs. 38.57±8.12; caspase-3 (fluorescence intensity): 6 hours: 0.37±0.07 vs. 0.69±0.23, 12 hours: 0.42±0.07 vs. 0.86±0.13, 24 hours: 0.53±0.11 vs. 1.36±0.23, 48 hours: 0.50±0.08 vs. 1.48±0.15, all P<0.05] in hydrogen treatment group. Hydrogen inhalation can down-regulate the systemic inflammatory response to ameliorate the intestinal injury, and it may improve the septic process and increase the survival rate of mice with severe sepsis.