Vacuolation Of Epithelial Cells Research Articles

The effects of saline water consumption on sperm parameters, testicular histopathology, hormonal and antioxidants concentrations in Barki Rams

The study aimed to assess the effects of water salinity on the sperm parameters, levels of cortisol, LH, FSH, testosterone and antioxidants as well as the testes’ histopathology in Barki rams. Fifteen healthy Barki rams (1–1.5 years) were divided into three equal depending on the type of drinking water for nine months. The rams in the tap water group (TW, water that contained 350 ppm of total dissolved salts (TDS). Males in the high saline water group (HSW) were permitted to consume high saline water with 8,934 ppm TDS, whereas those in the second group were permitted to have moderately saline water (MSW, 4,557 ppm TDS). High salt concentration in drinking water had adverse effect on sperm viability, morphology and sperm cell concertation. Nitric oxide and malondialdehyde concentrations in blood were significantly higher in the MSW and HSW groups than in TW. There was a significant decrease in glutathione concentration as well as superoxide dismutase activity in TDS and HSW. Cortisol was most highly concentrated in the HSW, next in the MSW, and least in TW. The testosterone, LH, and FSH concentrations in the HSW and MSW groups were significantly lower than in TW. As the salt concentration in drinking water increases, damage to testicular tissue. The MSW group demonstrating vacuolation of lining epithelial cells with pyknotic nuclei in the epididymis and necrosis and desquamation of spermatogenic cells in seminiferous tubules while HSW group displaying desquamated necrotic cells and giant cell formation in the epididymis, as well as damage to some of the seminiferous tubules and showed congestion, vacuolation of spermatogenic epithelium of seminiferous tubules, and desquamated necrotic spermatogenic epithelium. In conclusion, the salinity of the water has detrimental impacts on the sperm morphology, viability and concentration, hormones and antioxidant levels in Barki rams.

Adenomyotic Lesions Are Induced in the Mouse Uterus after Exposure to NSAID and EE2 Mixtures at Environmental Doses.

The aim of this study was to assess the long-term effect of exposure to environmentally relevant doses of non-steroidal anti-inflammatory drugs (NSAIDs; ibuprofen, and diclofenac) and 17β-ethinylestradiol (EE2) on the mouse uterus. NSAID-EE2 mixtures were administered in the drinking water from gestational day 8 until 8 weeks post-birth (i.e., during embryo development, lactation, puberty, and sexual maturity). The incidence of adenomyosis lesions (presence of endometrial glands in the inner myometrium) increased up to 60% in the uterus of 8-week-old exposed females (F1) and to 85% in F2 females (exposed father). Histological analysis revealed aberrant proliferation and apoptosis, vacuolization of epithelial cells, and increased incidence of abnormal glands in the luminal and glandular epithelium in F1 and F2 uteri. Moreover, myofibroblast proportion (alpha-smooth muscle actin (α-SMA) expression analysis) and collagen expression (Picrosirius red stain; a fibrosis hallmark) were increased in F1 and F2 endometrium. Connexin-43 was aberrantly distributed in the endometrial stroma and glands of F1 and F2 uteri. Conversely, uterine 17β-estradiol and progesterone levels were not affected in F1 and F2 females. These findings demonstrated that in mice, chronic exposure to NSAID and EE2 mixtures at environmental doses intergenerationally affects uterine physiology, particularly the endometrium. It may serve as a model to study the pathophysiology of human adenomyosis.

Effects of Tannic Acid Supplementation of a High-Carbohydrate Diet on the Growth, Serum Biochemical Parameters, Antioxidant Capacity, Digestive Enzyme Activity, and Liver and Intestinal Health of Largemouth Bass, Micropterus salmoides.

We investigated the effects of dietary tannic acid (TA) supplementation of a high-carbohydrate diet on growth, feed utilization, whole-body proximate composition, serum biochemical indicators, antioxidant capacity, digestive enzyme activity, and liver and intestinal health of juvenile largemouth bass, Micropterus salmoides (initial mean weight: 8.08 ± 0.08 g). Five diets were prepared, including a positive control (dietary carbohydrate level, 16%, LC0), a negative control (dietary carbohydrate level, 21%, HC0), and three TA-supplementation diets based on the negative control diet with TA addition at 200, 400, and 800 mg/kg, respectively. After 8 weeks of feeding, the results showed that compared with the LC0 diet, 400-800 mg/kg dietary TA significantly improved the survival rate of largemouth bass (P < 0.05) while significantly reducing its weight-gain rate and specific growth rate (P < 0.05). Compared with the HC0 diet, 400 mg/kg dietary TA significantly increased serum catalase activity (P < 0.05), and significantly decreased serum malondialdehyde, liver glycogen, lightness (L ∗), and yellowness (b ∗) (P < 0.05). Moreover, compared with the HC0 diet, 200-400 mg/kg dietary TA effectively improved the vacuolation of hepatocytes caused by the high-carbohydrate diet and reduced the occurrence of intestinal epithelial cell vacuolation and necrosis. In turn, 800 mg/kg dietary TA significantly inhibited protease activity in the pyloric caecum and intestine (P < 0.05). In conclusion, dietary supplementation with TA inhibited protease activity, which resulted in decreased growth performance in largemouth bass. However, it was also found that 200-400 mg/kg TA enhanced the antioxidant capacity of largemouth bass in the case of the high-carbohydrate diet, reduced liver glycogen levels, and improved liver and intestinal health. Finally, it should be noted that, when the dietary TA level exceeded 800 mg/kg, TA appeared to play a pro-oxidation role in the liver, which may cause oxidative stress in the liver.

Analysis of histopathology and changes of major cytokines in the lesions caused by Mycoplasma ovipneumoniae infection

BackgroundMycoplasma ovipneumoniae (M. ovipneumoniae) is one of the main pathogens of sheep pneumonia, causing a series of clinical symptoms, such as depression, anorexia, hyperthermia, cough, dyspnea, and tract secretions. In recent years, the prevalence of M. ovipneumoniae pneumonia has become increasingly serious in sheep farms in Ningxia, China, leading to the death of sheep, and causing significant economic losses. In this study, the pathological organs infected by M. ovipneumoniae were collected to observe histopathological change, to determine the tissue localization of M. ovipneumoniae, and to analyze the cytokine changes, which lays a basis for the diagnosis and pathogenesis of M. ovipneumoniae disease.ResultsIn this study, M. ovipneumoniae was detected in 97 of 105 samples collected from 13 large-scale sheep farms for nucleic acid by PCR. One representative isolate per farm was isolated from 13 farms. The lesions caused by M. ovipneumoniae were mainly in the trachea, bronchus, and lung, including necrosis of tracheal mucosal epithelial cells, disintegration of some epithelial cells, edema of mucosal lamina propria, with inflammatory cell infiltration, cytoplasmic vacuolization of epithelial cells of bronchial mucosa, massive infiltration of inflammatory cells in the alveolar space of lung, necrosis and hyperplasia of alveolar epithelial cells. Immunohistochemical analysis showed that the proportion of M. ovipneumoniae positive area in the lung was the largest, followed by that in the bronchus and trachea. Compared to healthy animals, diseased animals exhibited up-regulated gene expression levels of IL-1β, IL-6, and NF-κB in the trachea, bronchus, and lungs. In contrast, the expression of IL-10, IL-12, and IFN-γ was primarily limited to the trachea and bronchus. The expression of IL-1β showed differential patterns across different lung regions, with variations observed among lung lobes. Additionally, other cytokines consistently showed significant up-regulation specifically in the bronchus.ConclusionsM. ovipneumoniae is primarily found in the lungs of infected individuals. NF-κB, an essential transcription factor, is involved in the regulation of IL-1β transcription. IL-12 may enhance the cytotoxic function of natural killer cells during M. ovipneumoniae infection. Those findings demonstrate the distinct expression profiles of cytokines in various anatomical sites throughout disease progression, suggesting the potential role of bronchial tissue as a major site of immune response.

RNA-seq analysis revealed the pathogenicity of Vibrio vulnificus to American eel (Anguilla rostrata) and the strategy of host anti-V. vulnificus infection

Vibrio vulnificus is a commonly pathogenic bacterium in cultivated eels, but its pathogenicity to American eel (Anguilla rostrata) and the molecular mechanism of host anti-V. vulnificus infection remains uncertain. In this study, American eels were infected with different dose of V. vulnificus to determine the LD50. Then, bacterial load in the liver and kidney histopathology were assessed post the LD50 of V. vulnificus infection. Additionally, gene expressions of 18 immune related genes in the liver, spleen and kidney were detected. Furthermore, transcriptome sequencing and enrichment of differentially expressed genes (DEGs) were analyzed in the eel spleens between pre-infection (Con_0), post-36 h (Vv_36), and post-60 h (Vv_60) infection. The results showed that LD50 of V. vulnificus to American eels was determined to be 5.0 × 105 cfu/g body weight, and the bacterial load peaked at 24 and 12 h post the infection (hpi) in the kidney and liver, respectively. The histopathology was highlighted by necrotic hepatocytes and splenic cells, congestion blood vessels in liver and spleen, atrophied glomeruli and vacuolization of renal tubular epithelial cells. The results of RT-PCR revealed that 18 host immune-related genes showed significantly up or downregulated expression post-infection compare to that of pre-infection. Finally, results of the RNA-seq revealed 16 DEGs play essential role to the immunosuppression in American eels, and the protein-protein interactions shed light on the widespread upregulation GEGs related to metabolism and immune response maintained the host cell homeostasis post the V. vulnificus infection, shedding new light on our understanding of the V. vulnificus pathogenesis towards understudied American eel and the host anti-V. vulnificus infection strategies in gene transcript.

Effectivity test of ethanol extract of black pepper (Piper nigrum) against mortality of Aedes aegypti and its effect on midgut morphological changes

Aedes aegypti is the vector of Dengue Hemorrhagic Fever (DHF). Dengue vector management usually depends on synthetic larvicides. However, it can lead to insect resistance and leave chemical pollutant residues. Therefore natural larvicides are needed for their environmentally friendly and do not induce insect resistance. Piper nigrum fruit is expected to contain secondary metabolites beneficial as larvicides to Aedes aegypti. This study aims to test the effectivity of the ethanol extract of Piper nigrum fruit on the mortality of Aedes aegypti larvae and determine the effect of extract exposure on larval midgut morphological changes. The study used a completely randomized design with five experimental groups and four repetitions as follows: K (aquades + 0% extract), P1 (0.25% extract), P2 (0.50% extract), P3 (0.75% extract), P4 (1% extract). Data analysis used one-way ANOVA and continued to the LSD Post hoc test. The extract's effectiveness was analyzed using the probit test and LC50 confirmation. Changes in the morphology of larval midgut were observed descriptively and presented in the form of tables and figures. The results showed that the 1% treatment resulted in the highest larval mortality of 93% and the lowest in the 0.25% treatment of 43%. The LC50 value was 2,666 based on the probit analysis. The ethanol extract of Piper nigrum fruit affected midgut morphological changes of Aedes aegypti larvae characterized by peripheral membrane damage, microvilli thinning, epithelial cell swelling and vacuolation, and basement membrane damage.

Maturation of the medaka immune system depends on reciprocal interactions between the microbiota and the intestinal tract.

The interactions between the host immune system and intestinal microorganisms have been studied in many animals, including fish. However, a detailed analysis has not been performed in medaka, an established fish model for biological studies. Here, we investigated the effect of immunodeficiency on the microbiota composition and the effect of gut bacteria on intestinal epithelial development and immune responses in medaka. Chronological analysis of the intestinal microbiota of interleukin 2 receptor subunit gamma (il2rg) mutant medaka showed a gradual decrease in the evenness of operational taxonomic units, mainly caused by the increased abundance of the Aeromonadaceae family. Exposure of wild-type medaka to high doses of an intestine-derived opportunistic bacterium of the Aeromonadaceae family induced an inflammatory response, suggesting a harmful effect on adult il2rg mutants. In addition, we established germ-free conditions in larval medaka and observed large absorptive vacuoles in intestinal epithelial cells, indicating a block in epithelial maturation. Transcriptome analysis revealed a decrease in the expression of genes involved in the defense response, including the antimicrobial peptide gene hepcidin, whose expression is induced by lipopolysaccharide stimulation in normal larvae. These results show that reciprocal interactions between the microbiome and the intestinal tract are required for the maturation of the medaka immune system.

Polystyrene microplastics disrupted physical barriers, microbiota composition and immune responses in the cecum of developmental Japanese quails

Microplastics, a new type of emerging pollutant, is ubiquitous in terrestrial and water environments. Microplastics have become a growing concern due to their impacts on the environment, animal, and human health. Birds also suffer from microplastics contamination. In this study, we examined the toxic effects of polystyrene microplastics (PS-MPs) exposure on physical barrier, microbial community, and immune function in the cecum of a model bird species—Japanese quail (Coturnix japonica). The one-week-old birds were fed on environmentally relevant concentrations of 20 µg/kg, 400 µg/kg, and 8 mg/kg PS-MPs in the diet for 5 weeks. The results showed that microplastics could cause microstructural damages characterized by lamina propria damage and epithelial cell vacuolation and ultrastructural injuries including microvilli breakage and disarrangement as well as mitochondrial vacuolation in the cecum of quails. In particular, blurry tight junctions, wider desmosomes spacing, and gene expression alteration indicated cecal tight junction malfunction. Moreover, mucous layer breakdown and mucin decrease indicated that chemical barrier was disturbed by PS-MPs. PS-MPs also changed cecal microbial diversity. In addition, structural deformation of cecal tonsils and increasing proinflammatory cytokines suggested cecal immune disorder and inflammation responses by PS-MPs exposure. Our results suggested that microplastics negatively affected digestive system and might pose great health risks to terrestrial birds.

Antioxidation, immunity and hepatopancreatic histology of red swamp crayfish (Procambarus clarkii) subjected to low-temperature aerial exposure stress and re-immersion

Desiccation is a stressful situation that decapods often experience during live transportation. This study investigated the effects of low-temperature aerial exposures (LTAEs) (dry exposure (DL) and moist exposure (ML) at 6 °C) and re-immersion on the antioxidative and immune responses and hepatopancreatic histopathology in P. clarkii. Compared to the control group (normally feeding at 24.0 °C water temperature), the crayfish under LTAEs showed overall severe hepatopancreatic oxidative damage, with significantly increased malondialdehyde (MDA) contents and significantly reduced total antioxidant capacity (T-AOC), and oxidant damage was not fully recovered even after 12 h of re-immersion; the expression of hsp70 was significantly increased within 24–48 h of stress and re-immersion. The activity of hemolymphatic acid phosphatase (ACP) was significantly increased during 24–48 h of the stress and at 12 h of re-immersion; the activity of aspartic aminotransferase (AST) and alanine aminotransferase (ALT) was significantly increased throughout the experiment; and the gene expression of proPO or TLR was significantly increased during 12–48 h of the stress. Severe histopathological changes (lumen dilatation, vacuolation of epithelial cells and reduced cell numbers) were observed in hepatopancreas at 48 h of stress and 12 h of re-immersion. These results indicated that 48 h of low-temperature aerial exposure stress stimulated the non-specific immunity but adversely affected the antioxidation and hepatopancreatic histomorphology of P. clarkii, whereas 12 h of re-immersion was not sufficient to restore crayfish from stress to a normal state.

Renal toxicity caused by diethylene glycol: an overview.

Diethylene glycol (DEG) is nephrotoxic, potentially resulting in high morbidity and mortality. Its main nephrotoxic by-product is diglycolic acid (DGA). This narrative overview summarizes selected literature with a focus on clinical findings, pathophysiology, diagnosis including morphological features of renal biopsies, and management. The kidney injury in DEG poisoning is secondary to proximal tubular necrosis caused by DGA. Marked vacuolization and edema of epithelial cells obstruct the lumen, reducing urine flow and, consequently, resulting in anuria and uremia. The clinical alterations due to DEG poisoning are dose-dependent. Patients may present with gastrointestinal symptoms and anion gap metabolic acidosis, followed by renal failure, and, later, encephalopathy and neuropathy. Although this three-phase pattern has been described, signs and symptoms may be overlapping. Data about DEG intoxication is scarce. Sometimes the diagnosis is challenging. The management includes supportive care, gastric decontamination, correction of acid-base disorders, and hemodialysis. The understanding of the metabolic processes related to DEG poisoning may contribute to its management, preventing death, serious sequels, or irreversible lesions.

Histopathology, immunoenzyme activity and transcriptome analysis of immune response in silver pomfret infected by cryptokaryon (Cryptorchidism irritant)

Cryptorchidism irritant (CI) infection is a major problem in the culturing process of silver pomfret (Pampus argenteus), which can result in rapid and massive death. However, there is limited information available on the immune response of silver pomfret infected by CI. To address this gap, we sampled naturally infected fish and observed milky white translucent oval CI trophozoites on the gills, body surface, and fin rays. Histological analysis showed that CI infection led to vacuolation of epithelial cells and a decrease in blood cells in the gills. We also performed transcriptome profiling of the gill, kidney complex, and spleen, generating 399,616,194 clean reads that assembled into 101,228 unigenes, which were annotated based on public databases. We detected 14,369 differentially expressed genes, and selected several key immune-related genes for further validation using RT-qPCR. The Graft-versus-host pathway and Allograft rejection pathway were enriched in the gills, leading to inflammation and ulceration. CI infection activated the immune system, increasing levels of interleukin-1 beta and MHC class II antigen, and also activated innate and acquired immune genes in silver pomfret. Furthermore, we measured the activities of five immune-related enzymes (SOD, AKP, CAT, CSH and ACP), which all increased to varying degrees after CI infection. Our findings enhance our understanding of the immune response of fish to parasitic infection and may contribute to the development of strategies to prevent high mortality in CI-stimulated fish in aquaculture.

Wenzhou shrimp virus 8 (WzSV8) detection by unique inclusions in shrimp hepatopancreatic E-cells and by RT-PCR

The genome sequence of Wenzhou shrimp virus 8 (WzSV8) (GenBank record KX883984.1) was described in 2015 from wide screening for RNA viruses in aquatic animals. A closely related sequence (GenBank record OK662577.1) from the whiteleg shrimp Penaeus vannamei was deposited in 2021 under the name Penaeus vannamei picornavirus (PvPV). In 2022 another closely related sequence (GenBank accession: OP265432) was submitted under the name Penaeus vannamei solinvivirus (PvSV). In 2021, prior to the publication of PvPV and PvSV, we used an RT-PCR method devised from the sequence of KX883984.1 (described herein) to screen for WzSV8 in specimens of cultivated shrimp. Samples that gave positive RT-PCR results were subsequently tested by in situ hybridization (ISH) analysis to identify virus target tissues. Several tissues gave positive ISH results within morphologically normal nuclei. Thus, these tissues were of no use for diagnosis of WzSV8 by normal histological analysis. However, unique basophilic, cytoplasmic inclusions within vacuoles in tubule epithelial cells of the shrimp hepatopancreas (HP) were also found in the same WzSV8-positive shrimp specimens, sometimes accompanied by a smaller eosinophilic inclusion. We call these Lightner double Inclusions (LDI) that can be considered pathognomonic for diagnosis of WzSV8 infection when detected using the light microscope. Such abnormal WzSV8 inclusions could be seen scattered throughout the HP. However, the presence of vacuoles, and especially those containing viral inclusions, is highly abnormal in E-cells. Thus, focusing attention on the E-cell region of the HP for such features, serves as a useful approach in facilitating more rapid detection of WzSV8 infections in shrimp. Although no current proof of WzSV8 is the cause of disease, it is important to investigate new viruses and related tissue anomalies, even from normal cultivated shrimp, to determine whether they may have any relationship to significant negative effects on the production.

Histological study of the role of CD34+ stem cells and mast cells in cyclophosphamide-induced thymic injury in rats and the possible attenuating role of melatonin

Cyclophosphamide (CP) is an anticancer drug that adversely affects immunity and thymus structure. Melatonin is a hormone secreted by the pineal gland. It boosts immunity and has antioxidant properties. Therefore, the present study was conducted to investigate the possible protective effect of melatonin on CP-induced changes in the rat thymus. Forty male albino rats were used and divided equally into four main groups. Group I was the control group. Group II (melatonin group) received melatonin at a dose of 10 mg/kg body weight/day by intraperitoneal injection throughout the experimental period. Group III (CP group) received 200 mg/kg body weight CP by a single intraperitoneal injection. Group IV (CP + melatonin group) received melatonin intraperitoneally at a dose of 10 mg/kg body weight/day starting 5 days prior to CP injection until the end of the experiment. All rats were euthanized 7 days after CP injection. Administration of CP in group III resulted in depletion of the cortical thymoblasts. In addition, CD34-immunopositive stained stem cells decreased and mast cell infiltration increased. Electron microscopy showed degeneration of thymoblasts and vacuolization of epithelial reticular cells. Administration of melatonin with CP in group IV showed considerable protection of thymic histology. In conclusion, melatonin may protect against CP-induced thymic injury.

Histopathology of Nephrotoxicity Associated with Administered Water Extract Purple Sweet Potato (&lt;i&gt;Ipomoea batatas&lt;/i&gt;) in Mice (&lt;i&gt;Mus musculus&lt;/i&gt;) in Stratified Phases of Dose

The main aim of the registered purple sweet potato (Ipomoea batatas) is to provide minimize the adverse chemical drugs, in addition to its anti-inflammatory, antioxidant, and antimicrobial effects. Potentially adverse effects may be observed in laboratory animals in particular, the extent to which this administration can cause toxicity. This study aimed to examine the histopathology of nephrotoxicity associated with administered water extracts of purple sweet potato in mice with stratified doses. The study was conducted at the Biomedical Laboratory, Faculty of Medicine, Universitas Islam Bandung on September 2019. Female mice (Mus musculus) strain Swiss Webster, aged between 6–8 weeks weighing 25 to 30 g, were obtained from Biopharma Laboratory, Bandung. The animal was acclimatized for seven days before being administered water extract purple sweet potato: eleven mice, one control group, and ten treatment groups underwent toxicity doses of purple sweet potato water extract administration. Purple sweet potato variant of Ayamurasaki prepared in various oral doses. The results show in the control group there were no histopathological changes, but in the group administered water extract purple sweet potato from the first phase seems in a mild grade of macrophage accumulation, mild vacuolization of tubular epithelial cells, mild vascular dilatation, and mild hydrophilic degeneration. In the second phase, macrophage accumulation was visible in moderate grades. The LD50 of purple sweet potato extract is greater than 5,000 mg/kgBW. The findings of this study indicate that registration of purple sweet potato extract in confirmatory doses is safe to administer and did not exhibit any mortality. The toxicity test of purple sweet potato water extracts in the kidney exhibits minimal chemical effects.

WITHDRAWN: The effects of fumonisin B1 at the No Observed Adverse Effect Level (NOAEL) and 5-times above on the renal histology and lipidome of rats

Fumonisin B1 (FB1) mycotoxin was intraperitoneally (IP) administered at the No Observed Adverse Effect Level (NOAEL = 0.2 mg/kg BW/day as IP equivalent, "L") and 5-times above ("H") to male rats, in a controlled ("C"), 5-day study (n = 10/group, total n = 30). BW (bodyweight) of H rats decreased after day 4, kidney weight after 5 days. Renal histology revealed tubular epithelial desquamation, tubular dilatation, nuclear swelling, pale chromatin, cell vacuolation and casual karyopycnosis (H). Lipidomic analysis was performed with liquid chromatography - time-of-flight mass spectrometry (LC-TOF). Renal sphinganine (Sa) concentration increased 500 (L) to 1000-fold (H) and Sa-1-P to over 200 and 350-fold, respectively), with FB1 dose-dependence. Renal triacyclglycerols, diacylglycerols, ceramides and sphingomyelins were depleted, while cholesterol and cholesterol ester concentrations increased. Spearman correlation of free sphingoid bases (Sa, Sa-1-P, sphingosine (So) and So-1-P) was positive with histopathological damage severity, sphingomyelins and ceramides provided negative relationship (-0.78 and -0.8, resp.). Two-way cluster analysis and sparse partial least squares discriminant analysis (sPLS-DA) was used for experimental group classification. Fully effective group separation was achieved for ceramides, sphingomyelins and phosphatidyl-cholines, highlighting molecular species of possible diagnostic value. Lipidomic results highlight possible re-consideration of the NOAEL.

Effects of Suramin on Polycystic Kidney Disease in a Mouse Model of Polycystin-1 Deficiency.

The aberrant activation of the purinergic signaling pathway has been shown to promote cyst growth and fluid secretion in autosomal dominant polycystic kidney disease (ADPKD). Suramin is an anti-parasitic drug that has strong anti-purinergic properties. Whether suramin could have a therapeutic effect on ADPKD has not been fully investigated. We examined the effect of suramin on cyst progression in a Pkd1 microRNAs transgenic mouse model that presented stable Pkd1 knockdown and moderate disease progression. The Pkd1-deficient mice were treated with suramin (60 mg/kg) by intraperitoneal injection twice a week from postnatal days 35 to 90. Kidney-to-body weight ratios, cyst indices, and blood urea nitrogen (BUN) levels were measured. Cell proliferation and macrophage infiltration were determined by immunohistochemistry. The suramin-treated group had significantly lower renal cyst densities, cell proliferation, and macrophage infiltration compared with saline-treated controls. Suramin significantly inhibited ERK phosphorylation and the expression of Il1b, Il6, Nlrp3, Tgfb, Fn1, P2rx7, and P2ry2 mRNAs in the kidneys. However, BUN levels remained high despite the reduction in cyst growth. Furthermore, plasma cystatin C and neutrophil gelatinase-associated lipocalin (NGAL) levels were significantly higher in the suramin-treated group compared with the control group. Periodic acid-Schiff staining revealed degenerative changes and epithelial cell vacuolation in the non-cystic renal tubules, which indicated phospholipidosis following suramin treatment. These results suggest that suramin may reduce renal cyst growth and inflammation, but the associated tubular cell injuries could limit its therapeutic potential. Other purinergic receptor antagonists with less nephrotoxicity may deserve further investigation for the treatment of ADPKD.

Effect of Aerobic Treadmill Exercise Regulating Glucose Transporter 9 on Renal Function in Hyperuricemia Rats

<bold>Objective</bold> To explore the possible mechanism of aerobic treadmill exercise in improving renal function in hyperuricemia (HUA) rats. <bold>Methods</bold> A total of 24 male SD rats were randomly divided into the Control group, the Model group and the Exercise group according to the random number table method, with 8 rats in each group. The Control group was given 2 g/(kg·d) distilled water intragastric administration, the Model group and the Exercise group were given 2 g/(kg·d) oxonic aid potassium salt (OAPS) intragastric administration, continuous intragastric administration for 10 days. From the 11th day, the Control group was given distilled water intragastric administration every other day, and the Model group was given OAPS intragastric administration every other day to maintain high uric acid status, with a dose of 2 g/(kg·d). In both groups, diet and water were not restricted, exercise freely, without other intervention, lasted for three weeks. The Exercise group was given intragastric administration as same as the Model group, and was given aerobic treadmill training simultaneously, the speed was 12 m/min, 20 minutes a time, once a day, continuous training for six days and rest for one day, continuous training for three weeks. Hematoxylin-eosin (HE) was used to stain the renal histopathological section, and the microscope (×400) was used to observe the structural changes of renal tubules and glomerulus; the peroxidase method was used to detect the renal function indexes such as uric acid (UA), creatinine (Cre) and urea nitrogen (BUN); enzyme-linked immunosorbent assay (ELISA) method was used to determine the contents of interleukin-1β (IL-1β) in serum and kidney tissues; Western blot was used to detect the expressions of Caspase-1, IL-1β and glucose transporter 9 (GLUT-9) in the kidney tissues. <bold>Results</bold> ① Pathological examination results: there was not obvious pathological changes were observed in the kidney tissues of the Control group; in the Model group, the renal tissue capsule was incomplete, the glomerulus was deformed and atrophic, and the renal tubules were disordered; in the Exercise group, the number of inflammatory cell infiltration was significantly decreased, the renal tubular structure was relatively intact, and the pathological changes such as cytoplasmic vacuolation of renal tubular epithelial cells were significantly decreased. ② UA, Cre and BUN content of serum: compared with the Control group, UA, BUN and Cre content in serum of the Model group and Exercise group was significantly increased (<italic>P</italic>&lt;0.05); compared with the Model group, UA, BUN and Cre content in serum of the Exercise group were significantly decreased (<italic>P</italic>&lt;0.05). ③ Serum and kidney IL-1β content: compared with the Control group, the expression of IL-1β in serum and kidney of the Model group and the Exercise group were significantly increased (<italic>P</italic>&lt;0.05); compared with the Model group, the expression of IL-1β in serum and kidney of the Exercise group was significantly decreased (<italic>P</italic>&lt;0.05). ④ Expressions of Caspase-1, IL-1β and GLUT-9 protein: compared with the Control group, the expressions of Caspase-1, IL-1β and GLUT-9 protein in kidney tissues of the Model group and the Exercise group were significantly increased (<italic>P</italic>&lt;0.05); compared with the Model group, the expressions of Caspase-1, IL-1β, and GLUT-9 protein in kidney tissues of the Exercise group were significantly decreased (<italic>P</italic>&lt;0.05). <bold>Conclusion</bold> Aerobic treadmill exercise can effectively reduce the level of uric acid and inflammatory response of hyperuricemia rats, and improve renal function, which may be related to the regulation of GLUT-9 protein expression.

Sperm Quality and Histology of the Testis and Epididymis in Chemical Castrated Male Cats with Intra-testicular Injection of Sodium Chloride Solution

The high population of feral cats in Bali, Indonesia, could be a problem associated with cat welfare and the risk of zoonotic diseases. Gonadectomy or male castration is an option for population control; however, there would be more economical ways to reach this goal. Sterilization using chemicals is an alternative to the surgical method of castration as it is reasonably priced, activated in a short time, and does not affect animal activity after the chemical administration. The present study aimed to evaluate intratesticular injection of Sodium chloride as an agent of chemosterilants in male cats. In the current study, 16 healthy male cats aged nearly one year old were randomly selected. The male cats were allocated into four groups containing four cats in each. Group 1 (control) received bilateral intratesticular injection with 0.25 mL normal saline solution containing 2% Lidocaine. The cats in groups 2, 3, and 4 received bilateral intratesticular injection of 0.25 mL Sodium chloride solution containing 2% Lidocaine with a concentration of 15%, 20%, and 25%, respectively. At the end of the study (after 30 days of injection), the cats were castrated and their testes were removed for sperm quality and histopathological evaluation. The results of the present study indicated that intratesticular injection of Sodium chloride significantly decreased the motility and viability rates, and increased the abnormalities of the sperms. Histopathological studies revealed marked depletion of spermatozoa in the testes and seminiferous tubular degeneration, seminiferous tubular atrophy, and epithelial cell vacuolation. In conclusion, the injection of 20-25% solution of Sodium chloride in testes had the potential as a chemosterilant for chemical sterilization in cats.

Subchronic oral exposure of tungsten induces myofibroblast transformation and various markers of kidney fibrosis.

Tungsten is a naturally occurring transition element used in a broad range of applications. As a result of its extensive use, we are increasingly exposed to tungsten from our environment, including potable water, since tungsten can become bioaccessible in ground sources. The kidneys are particularly susceptible to tungsten exposure as this is the main site for tungsten excretion. In this study, we investigated the prolonged effects of tungsten on the kidneys and how this may impact injury and function. When mice were exposed to tungsten in their drinking water for 1 mo, kidney function had not significantly changed. Following 3-mo exposure, mice were presented with deterioration in kidney function as determined by serum and urine creatinine levels. During 3 mo of tungsten exposure, murine kidneys demonstrated significant increases in the myofibroblast marker α-smooth muscle actin (αSMA) and extracellular matrix products: fibronectin, collagen, and matricellular proteins. In addition, Masson's trichrome and hematoxylin-eosin (H&E) staining revealed an increase in fibrotic tissue and vacuolization of tubular epithelial cells, respectively, from kidneys of tungsten-treated mice, indicative of renal injury. In vitro treatment of kidney fibroblasts with tungsten led to increased proliferation and upregulation of transforming growth factor β1 (TGFβ1), which was consistent with the appearance of fibroblast-to-myofibroblast transition (FMT) markers. Our data suggest that continuous exposure to tungsten impairs kidney function that may lead to the development of chronic kidney disease (CKD).

THE STATE OF MICROBIOTS AND MORPHOLOGICAL CHANGES OF THE SMALL INTESTINE IN EXPERIMENTAL ACUTE GENERALIZED PERITONITIS ON THE BACKGROUND OF STREPTOSOTOCIN-INDUCED DIABETES MELLITUS

The aim of the work: to analyze the microbiota and to study the morphological structure of the small intestine in the dynamics of acute peritonitis on the background of streptozotocin-induced diabetes mellitus.&#x0D; Materials and methods. Experimental researches were performed on 52 adult white male rats. The main group of animals was divided into three subgroups, on which bacteriological examination of parietal intestinal biotope and morphological examination of the small intestine on the first, third and seventh days of acute peritonitis on the background of concomitant diabetes were carried out. Diabetes mellitus was simulated by a single intraperitoneal injection of streptozotocin (60 mg / kg). On day 14 of diabetes mellitus, rats were injected with 0.5 ml of 10 % filtered fecal suspension into the abdominal cavity and acute generalized peritonitis was initiated. The control group consisted of rats (n=8), which were injected subcutaneously with saline 0.9 % NaCl.&#x0D; Results and discussion. Bacteriological examination of the parietal intestinal biotope and morphological examination of the small intestine were performed. In the first subgroup of animals with acute peritonitis on the background of streptozotocin-induced diabetes mellitus, the dominant microbiota isolated from the parietal intestinal habitat was represented by Escherichia coli (70 %), Candida spp. (20 %) and Staphylococcus spp. (20 %). A monoculture of microorganisms was identified in 40 % of animals. In the second subgroup, the number of microbial associations increased to 78 %. Three-component associations predominated in the third subgroup, dominated by Escherichia coli (57 %) and Enterobacter aerogenes (43 %). In the control group of animals Bifidibacterium spp., Lactobacillus spp., Escherichia coli, Staphylococcus spp were found. The progression of the inflammatory process on the background of hyperglycemia was accompanied by the increase in the number of Enterobacter aerogenes, Bacteroides spp, Candida spp. and by the decrease in the number of Escherichia coli. The results of morphological examination of the mucous membrane of the small intestine of rats at different stages of development of this combined pathology indicate the development of an active inflammatory reaction, which is the basis for the deterioration of absorption function and enteral insufficiency syndrome. Alterative changes manifested by uneven vacuolation of villi epithelial cells, areas of epithelial desquamation, as well as severe infiltration of the mucous membrane by neutrophilic granulocytes, monocytes and lymphocytes increased throughout the experiment and were most characteristic of animals of the third subgroup, mortality in which reached 38 %.&#x0D; Conclusions. The results of bacteriological examination of the parietal intestinal biotope of animals with acute peritonitis on the background of streptozotocin-induced diabetes mellitus indicate the dominance of Escherichia coli (65 %), Enterobacter aerogenes (23 %), Candida spp. (23 %), Bacteroides spp (19 %) among the representatives of the microbiota of the small intestine.&#x0D; The development of the inflammatory process of the peritoneum on the background of hyperglycemia is accompanied by a decrease in the height and thickness of the villi, crypt depth, dystrophic changes of mucosal cells, which indicates the decrease in the barrier function of the small intestine and is the cause of bacterial translocation, which is enhanced by qualitative and quantitative changes in microbes (predominance of associations of pathogens of aerobic gram-negative microorganisms and fungi of the genus Candida).