Severe Histopathological Changes Research Articles

Investigation of the protective effect of rosmarinic acid in rats given high dose gentamicin

Abstract
 Purpose: Gentamicin is an antibiotic, used to treat infections caused by microorganisms. Endogenous antioxidants are strengthened by using antioxidant protectors against the toxic effects caused by drugs. Rosmarinic acid has antioxidative, anti-inflammatory, antiapoptotic, and antitumoral effects. In our study, we aimed to investigate the protective effect, immunohistochemical changes, oxidative markers, histopathological changes and inflammation related Ifi44 expression of rosmarinic acid against gentamicin induced nephrotoxicity in rats.
 Materials and methods: In our study, 32 rats randomly divided into four groups. These groups are consisted of control group, gentamicin group received gentamicin 100 mg/kg/day, gentamicin + rosmarinic group acid received gentamicin 100 mg/kg/day and rosmarinic acid 50 mg/kg/day, and rosmarinic acid group received rosmarinic acid 50 mg/kg/day. At the end of the study, histopathological, immunohistochemical, and biochemical changes in kidney tissues evaluated. 
 Results: Rosmarinic acid reduced creatine, urea, blood urea nitrogen, and total oxidative stress in blood serum. The toxic effect of gentamicin caused severe histopathological changes in the kidneys. A slight decrease in histopathological changes observed in the gentamicin + rosmarinic acid group. Antiproliferative Ifi44 expression was higher in the gentamicin group and gentamicin + rosmarinic acid group.
 Conclusion: As a result of the application of rosmarinic acid, a decrease in oxygen radicals and an increase in antioxidant levels observed. When used in combination with gentamicin and rosmarinic acid, the protective effect of rosmarinic acid was partially observed, but it could not provide full protection.

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.

Histopathology and virulence of an in vitro-adapted Trypanosoma evansi TEDC 953 strain (Thailand isolate) in mice.

Trypanosoma evansi is a blood and tissue protozoan parasite affecting domestic and wild animals. The T. evansi Thai strain, namely, T. evansi from dairy cattle number 953 (TEDC 953) strain, has been successfully isolated from dairy cattle and cultivated in vitro. The in vitro-cultivated parasite is useful for biological studies, evaluation of novel chemotherapeutic agents, and production of antigens for diagnostic tests. This study aimed to observe the histopathology and virulence of an in vitro-adapted T. evansi TEDC 953 strain in vivo. The histopathology and virulence of the TEDC 953 strain were clarified in mice. Six mice were infected with 1 × 105 trypomastigotes of TEDC 953 strain intraperitoneally, and four mice were in the negative control. Parasitemia was monitored daily, and the mice were euthanized on 30 days post-infection (DPI). Internal organs were collected for histopathological examination using hematoxylin and eosin staining. Histopathological lesions were found in the liver, lung, heart, kidney, spleen, and brain of the inoculated mice. The main histopathological feature was lymphoplasmacytic inflammation in parenchyma and perivascular areas of multiple organs, and the severity of histopathological changes was related to the presence of trypomastigotes in the regional vessels. Granulomatous inflammation was seen in meninges, pleura, renal capsule, renal pelvis, and spleen of some infected mice. Four mice died at 17, 24, 26, and 27 DPI with an average parasitemia of 4.05 × 1011 trypomastigotes/mL. The average survival time was 23.5 DPI (mice = 4). This study confirmed that the TEDC 953 strain is infectious and pathogenic in mice after the continuously cultivated in vitro. To replace the use of experimental animals, the in vitro-cultivated parasite can be used instead in further studies.

Derrone Targeting the TGF Type 1 Receptor Kinase Improves Bleomycin-Mediated Pulmonary Fibrosis through Inhibition of Smad Signaling Pathway.

Transforming growth factor-β (TGF-β) has a strong impact on the pathogenesis of pulmonary fibrosis. Therefore, in this study, we investigated whether derrone promotes anti-fibrotic effects on TGF-β1-stimulated MRC-5 lung fibroblast cells and bleomycin-induced lung fibrosis. Long-term treatment with high concentrations of derrone increased the cytotoxicity of MRC-5 cells; however, substantial cell death was not observed at low concentrations of derrone (below 0.05 μg/mL) during a three-day treatment. In addition, derrone significantly decreased the expressions of TGF-β1, fibronectin, elastin, and collagen1α1, and these decreases were accompanied by downregulation of α-SMA expression in TGF-β1-stimulated MRC-5 cells. Severe fibrotic histopathological changes in infiltration, alveolar congestion, and alveolar wall thickness were observed in bleomycin-treated mice; however, derrone supplementation significantly reduced these histological deformations. In addition, intratracheal administration of bleomycin resulted in lung collagen accumulation and high expression of α-SMA and fibrotic genes-including TGF-β1, fibronectin, elastin, and collagen1α1-in the lungs. However, fibrotic severity in intranasal derrone-administrated mice was significantly less than that of bleomycin-administered mice. Molecular docking predicted that derrone potently fits into the ATP-binding pocket of the TGF-β receptor type 1 kinase domain with stronger binding scores than ATP. Additionally, derrone inhibited TGF-β1-induced phosphorylation and nuclear translocations of Smad2/3. Overall, derrone significantly attenuated TGF-β1-stimulated lung inflammation in vitro and bleomycin-induced lung fibrosis in a murine model, indicating that derrone may be a promising candidate for preventing pulmonary fibrosis.

Evaluating the histomorphological and biochemical changes induced by Tributyltin Chloride on pituitary-testicular axis of adult albino rats and the possible ameliorative role of hesperidin

ABSTRACT This study was performed to explore in detail the toxic effects of Tributyltin Chloride (TBT) on the pituitary-testicular axis and the possible amelioration with Hesperidin. Seventy-two adult male albino rats were divided into four groups: Control group (I), TBT-treated group (II), TBT+Hesperidin group (III), and Recovery group (IV). Body and testicular weights were measured. Blood samples were taken to estimate serum levels of testosterone, FSH and LH hormones by enzyme-linked immunosorbent assay (ELISA). Malondialdehyde (MDA) level was measured in testes homogenates. Tissue samples from the pituitary glands and testes were processed for light, electron microscope examination, and immunohistochemical detection of anti-FSH, and Ki67 proteins. Results showed a statistically significant decrease in testicular weight, serum testosterone, FSH and LH levels and a significant increase in tissue MDA in the TBT group when compared to the control group. TBT treatment caused severe histopathological changes with decreased area percent of PAS-stained basophils, and anti FSH immuno-stained gonadotrophs in the pituitary gland. The testes of group II also showed marked tissue damage, cell loss with decreased epithelial height and decreased number of proliferating spermatogenic cells. Hesperidin supplementation with TBT proved significant amelioration of the previously mentioned parameters in both glands which could improve male fertility. In conclusion: The flavonoid Hesperidin has the potential to protect against the reproductive damage induced by TBT in susceptible individuals.

Melatonin pretreatment can improve the therapeutic effect of adipose-derived stem cells on CCl4-induced liver fibrosis

In this study, the therapeutic effect of Mel-incubated Adipose-derived mesenchymal stem cells (ADSCs) on CCl4-induced hepatic fibrosis was investigated. Mice with hepatic fibrosis were intraperitoneally injected with 8% CCl4 twice a week for 4 weeks. Starting from week 5, mice in the ADSC group and ADSC + Mel group were injected with 1 mL PBS cell suspension containing 1 × 106 ADSCs or ADSCs pretreated with 50 nM Mel twice a week for 2 consecutive weeks. Results: In the model group, severe histopathological changes were observed in the liver, including severe vacuolation, nuclear fragmentation, and liver fibrosis, and these changes were ameliorated by Mel-pretreated ADSCs. RT-qPCR results showed that Mel-induced ADSCs significantly inhibited the expression of proapoptotic genes (Caspase-8, Bax, and Caspase-3) and promoted the expression of an anti-apoptotic gene (Bcl-2). Immunohistochemical results showed that a large number of MMP-9-, TGF-β-, and MMP-2-positive cells were present in the liver tissues of the model group, while the number of positive cells was reduced by Mel-induced ADSCs. ELISA results showed that the ADSC pretreated with Mel also significantly reduced the expression levels of TNF-α and IL-6. Conclusion: ADSCs pretreated with Mel significantly improved CCl4-induced liver fibrosis.

Evaluation of the Effects of a 50 Hz Electric Field on Brain Tissue by Immunohistochemical Method, and on Blood Tissue by Biochemical, Physiological and Comet Method

The aim of this study was to evaluate the possible effects of a 50 Hz electric field on brain tissue and the positive effects of juglone (5-hydroxy-1,4-naphthoquinone) antioxidant activity, using the immunohistochemical technique on male Wistar-Albino rats. The effects on blood tissue were also examined using biochemical, physiological and comet methods. Animals were randomly divided into three groups (eight in each group): group I: control, group II: electric field, group III: 50 Hz electric field + juglone (5-hydroxy-1,4-naphthoquinone)/300 ppm. Juglone was applied per day by gavage over 30 days. At the end of the experimental procedure, animals were sacrificed and brain tissue was subjected to routine histologic and immunohistochemical processes. As a result of histophatological examination, the brain tissue of rats with 50 Hz electric field exposure showed severe histopathological changes. The differences between groups were statistically significant according to total comet score (p = 0.001). For the antioxidant parameters on the blood, SOD activity in the electric field group was significantly higher among the other groups, although we did not find significant differences in MDA, CAT activity level.

Effects of Single and Combined Exposure to Environmentally Relevant Concentrations of Cyfluthrin and Copper on Digestive Gland Histology of Lymnaea stagnalis

Pyrethroid insecticides and heavy metals frequently co-exist in aquatic systems, due to intensive anthropogenic activities, and their effects on aquatic organisms are needed to be investigated. In the present work, single and combined effects of environmentally realistic concentrations of cyfluthrin and copper on the digestive gland histology of the freshwater pond snail (Lymnaea stagnalis) were evaluated. Snails were exposed to 1 µg/L cyfluthrin, 10 µg/L copper, and 1 µg/L cyfluthrin + 10 µg/L copper mixture for 96 h. Cyfluthrin-exposed samples showed apparent vacuolization, basal lamina separations, and disrupted digestive cells. Exposure to copper alone gave rise to enlargements of the tubule lumens and the intertubular area, degenerated tubules, atrophied basophilic cells, prominently disrupted and ruptured digestive cells, and nuclear enlargements in some basophilic cells. Cyfluthrin and copper mixture caused more severe histopathological changes in the digestive gland. General tissue appearance was altered by prominently degenerated, and fused tubules lacking cellular structure; tubule lumens filled with cellular content were noted. Increased intertubular regions were filled with connective tissue formations. Progressive disruption of digestive cells; and basophilic cell atrophy were also observed. The findings of the current study highlighted that cyfluthrin and copper at environmentally relevant concentrations caused a biological response in L. stagnalis; however, this response was more intense following their co-exposure.

Immunopathological mechanisms in the early stage of Mycobacterium avium subsp. paratuberculosis infection via different administration routes in a murine model.

Mycobacterium avium subspecies paratuberculosis (MAP) is the causative agent of Johne's disease, a chronic emaciating disease of ruminants that causes enormous economic losses to the bovine industry, globally. However, there are still remaining clues to be solved in the pathogenesis and diagnosis of the disease. Therefore, an in vivo murine experimental model was tried to understand responses in early stage of MAP infection by oral and intraperitoneal (IP) routes. In the MAP infection size, and weight of spleen and liver were increased in the IP group compared with oral groups. Severe histopathological changes were also observed in the spleen and liver of IP infected mice at 12 weeks post-infection (PI). Acid-fast bacterial burden in the organs was closely related to histopathological lesions. In the cytokine production from splenocytes of MAP-infected mice, higher amounts of in TNF-α, IL-10, and IFN-γ were produced at early stage of IP-infected mice while IL-17 production was different at time and infected groups. This phenomenon may indicate the immune shift from Th1 to Th17 through the time course of MAP infection. Systemic and local responses in the MAP-infection were analyzed by using transcriptomic analysis in the spleens and mesenteric lymph nodes (MLN). Based on the analysis of biological processes at 6 weeks PI in spleen and MLN in each infection group, canonical pathways were analyzed with ingenuity pathway analysis in the immune responses and metabolism especially lipid metabolism. Infected host cells with MAP increased in the production of proinflammatory cytokines and reduced the availability of glucose at early stage of infection (p < 0.05). Also, host cells secreted cholesterol through cholesterol efflux to disturb energy source of MAP. These results reveal immunopathological and metabolic responses in the early stage of MAP infection through the development of a murine model.

RELATIONSHIP OF HELICOBACTER PYLORI CAGA AND VACA STATUS TO MORPHOLOGICAL CHANGES OF GASTRIC MUCOSA AND PRIMARY CLARITHROMYCIN RESISTANCE RATE IN PATIENTS WITH CHRONIC GASTRITIS: A CROSS-SECTIONAL STUDY.

The aim: To investigate the relation of H. pylori CagA and VacA status to morphological changes of gastric mucosa and primary clarithromycin resistance rate in patients with chronic gastritis. Materials and methods: A cross-sectional study was conducted between May 2021 and January 2023, involving 64 patients with H. pylori-associated chronić gastritis. The patients were assigned to two groups according to the H. pylori virulence factors (CagA and VacA) status. The grades of inflammation, activity, atrophy, and metaplasia were determined according to the Houston-updated Sydney system. The identification of H. pylori genetic markers of antibiotic resistance and pathogenicity was performed by the polymerase chain reaction using paraffin stomach biopsies. Results: Patients with CagA- and VacA-positive H. pylori strains had significantly higher grades of inflammation both in the antrum and in the corpus of the stomach, activity of gastritis in the antrum, higher incidence and grade of atrophy in the antrum. Primary resistance to clarithromycin was significantly more prevalent in patients with CagA- and VacA-negative H. pylori strains (58.3% vs. 11.5%, p=0.002). Conclusions: Positive CagA and VacA status is related to more severe histopathological changes of gastric mucosa. In contrast, the rate of primary clarithromycin resistance is higher in patients CagA- and VacA-negative H. pylori strains.

Histopathological and immunohistochemical studies on the influence of orally administration monosodium glutamate, a food additive dependent on time in vivo

BackgroundMonosodium glutamate is a widely used flavor enhancer/food additive in meat, fish, milk and cheese or vegetable origins. Our present study aimed to assess the effect of utilization monosodium glutamate on the cardiac, splenic, hepatic and renal organs in mice. Thirty animals were divided into three groups: control group; glutamate 10 treated groups and glutamate 15 treated group. The experimental period was two successive weeks.ResultsOral administrations of monosodium glutamate for 10 days induced moderate histopathological changes in cardiac, splenic, hepatic and renal tissues and also resulted in significant increase of the nuclear factor-kB expression depend on time of administration as compared to the control group, while treatment with monosodium glutamate for 14 days resulted in severe histopathological changes as well as highly significant increase of the nuclear factor-kB expression as compared to the control group.ConclusionsIt could be concluded that oral administration of monosodium glutamate induced histopathological alterations, and cytotoxicity in cardiac, splenic, hepatic and renal tissues dependent on time.

Histopathological and Biochemical Evaluation of Albendazole in the Treatment of Infected Mice with Hydatid Cyst

Introduction: Hydatidosis, caused by the larval stage of Echinococcus granulosus, is a prevalent parasitic disease affecting both humans and animals. Albendazole is currently the most effective drug for treating hydatid cysts. This research aimed to investigate the histopathological and biochemical effects of Albendazole on the liver, lung, and kidney of mice experimentally infected by hydatid cysts.&#x0D; Materials and methods: A total of 20 mice weighing approximately 220 g were used. The rats were randomly divided into the Albendazole group (100 mg/kg/day) and the control group (infected Rats without treatment). At the end of the experiment, tissue samples from the liver, lung, and kidney were collected for histopathological evaluation. Liver blood tests were used to assess liver functions or liver injury (alkaline phosphatase, alanine aminotransferase, and bilirubin).&#x0D; Results: After 30 days of daily treatment, the total numbers of cysts, size, and weight of the largest cyst were significantly lower in the Albendazole group, compared to the control group. The study addressed histopathological changes in the liver, kidneys, and lungs caused by hydatid cysts, such as tissue necrosis, hemorrhage, and local inflammation, indicating the potential for serious complications and significant damage to these organs. The group treated with Albendazole showed severe histopathological changes in the liver, kidneys, and lungs, compared to the control group. This suggests that Albendazole may trigger a more aggressive response in these organs to the cysts, leading to increased tissue damage. In addition, alkaline phosphatase, alanine aminotransferase, and bilirubin concentrations revealed a significant increase in the Albendazole group.&#x0D; Conclusion: While Albendazole is an effective drug for treating hydatidosis, it can also cause severe side effects on various organs in the body. Therefore, alternative treatment strategies need to be developed to minimize these adverse effects.

Hepatotoxicity of Short Term Exposure to Mancozeb Fungicide in Male Wistar Rats

Mancozeb is a dithiocarbamate fungicide used effectively to protect plant products against fungi. The hepatic effects of short term exposure to mancozeb in adult male Wistar rats were investigated in the present study. Twenty-four animals were divided into four equal groups. Two groups were administered mancozeb (60 mg/kg body weight as single dose or 30 mg/kg body weight daily for 10 days, intraperitoneally), and the others, which served as control groups, received normal saline. Liver biochemical parameters in plasma were measured using standard methods. Liver homogenates were analysed for oxidative stress biomarkers and liver histopathology was studied. Single dose and 10 days exposures of mancozeb caused elevation in the activities of Alanine Transaminase (ALT), Aspartate Transaminase (AST), Alkaline Phosphatase (ALP), Lactate Dehydrogenase (LDH), and Gamma Glutamyl Transpeptidase (GGT) in plasma (p&lt;0.05-0.001) compared with control. Mancozeb also caused elevation in the plasma level of total bilirubin, and reductions in albumin, total protein, and conjugated bilirubin. In addition, Malondialdehyde (MDA) and Advanced Oxidation Protein Product (AOPP) levels were increased in hepatic tissues (p&lt;0.001) of all mancozeb exposed rats. Furthermore, hepatic levels of protein, reduced Glutathione (GSH) and vitamin C were decreased (p&lt;0.01), together with the activities of Superoxide Dismutase (SOD), catalase, and Glutathione Peroxidase (GPx) enzymes (p&lt;0.01-0.001). Histological analysis showed severe histopathological changes in mancozeb exposed rats. The results demonstrated that single dose intraperitoneal exposure of mancozeb (60 mg/kg body weight) or short term (10 days) daily exposure at 30 mg/kg body weight is capable of causing hepatotoxic effects in rats.

Protective Role of Quercetin on Thioacetamide-Induced Renal Injury in Adult Male Albino Rats

Background: Thioacetamide (TAA) is a recognized industrial poisonous agent drastically used in animal studies for induction of hepatic necrosis, fibrosis and cirrhosis. It is additionally reported to be nephrotoxic through induction of oxidative stress. Quercetin (QE) has a high antioxidant capacity via free radical scavenging, transition metal ion binding, and lipid peroxidation inhibition. Aim of the work: The goal of this research was to see if QE may help reduce the negative effects of thioacetamide on renal tissue by histological examination of the kidney. Material and methods: Twenty four adult male Albino rats 7–9 weeks old around 180–200g body weight were allocated into 3 groups; Group I (n=8) receiving distilled water, Group II (n=8); rats receiving TAA and Group III (n=8); receiving TAA and QE. Results: Renal tissues were examined under a light microscope stained by Harris Hematoxylin &amp; Eosin (H&amp;E stain), Periodiceeacid Schiff (PAS) andjMasson’srTrichrome for TAA-treated groups revealed severe histopathological changes, whereas specimens obtained from QE-treated groups showed onlywmild changes. Immunohistochemical results corroborated these findings. Conclusion: This study demonstrated the ameliorative consequences of QE in opposition to TAA-induced renal injury in rats. The result of this study might contribute in the development of a novel complementary alternative medication in combating and therapeutic intervention of TAA-induced renal injury. KEY WORDS: Thioacetamide, renal, quercetin, albino rat and histopathology.

Melatonin ameliorates the adrenal and pancreatic alterations in streptozotocin-induced diabetic rats: Clinical, biochemical, and descriptive histopathological studies.

Previous studies have demonstrated the beneficial effects of melatonin in diabetic rats. However, limited studies have been conducted on the potential effects of melatonin on the descriptive histopathological and morphometric findings in different compartments of the adrenal glands in diabetic animal models. In this study, using a streptozotocin (STZ)-induced diabetic rat model, we sought to examine histological alterations in the pancreas and adrenal glands and observe the effect of the administration of melatonin on the histopathology and morphology of the pancreas and the adrenal gland cortex and medulla that are altered by STZ-induced hyperglycemia. Rats were randomly assigned to four different groups: Group I, normal control; Group II, melatonin group (MT) (10 mg/kg/day); Group III, (diabetic STZ group), and Group IV, diabetic (STZ) + melatonin group (MT). Throughout the experiment, the animals' fasting blood sugar levels were measured. Blood was obtained to determine the animals' cumulative blood sugar levels after sacrification. For histological and morphometrical evaluations, the pancreatic and adrenal gland tissues were dissected and processed. Our results showed that diabetic rats receiving melatonin significantly (P < 0.05) improved their fasting blood sugar and cumulative blood sugar levels compared to the diabetic group not receiving melatonin. Furthermore, histopathological examinations of the pancreatic and adrenal tissues of the diabetic rats indicated the occurrence of severe histopathological and morphometric changes. Morphometric analysis of the adrenals indicated a significant increase (P < 0.05) in the thickness of the cortex zones [zona glomerulosa (ZG), zona fasciculata (ZF), and zona reticularis (ZR)] for the diabetic STZ group compared with other groups, and a significant decrease (P < 0.05) in the diameter of the in adrenal gland medullas in the diabetic STZ rats compared to the other groups. Furthermore, treatment with melatonin restored these changes in both the pancreatic and adrenal gland tissues and produced a significant (P < 0.05) improvement in the cortex and medulla thickness compared to the untreated diabetic rats. Overall, melatonin significantly reduced the hyperglycemic levels of glucose in diabetic rats and reversed the majority of histopathological alterations in the tissues of the pancreas and adrenals, demonstrating its anti-diabetic activity.

Anti-Inflammatory and Analgesic Effects of Curcumin Nanoparticles Associated with Diclofenac Sodium in Experimental Acute Inflammation.

The present study evaluated the anti-inflammatory and analgesic effects of conventional curcumin (cC) and curcumin nanoparticles (nC) associated with diclofenac sodium (D) in experimental acute inflammation (AI) induced by carrageenan administration. Seven groups of eight randomly selected Wistar-Bratislava white rats were evaluated. One group was the control (C), and AI was induced in the other six groups. The AI group was treated with saline solution, the AID group was treated with D, the AIcC200 and AInC200 groups were treated with cC and nC, respectively, while AIcC200D and AInC200D were treated with cC and nC, respectively, both associated with D. Conventional curcumin, nC, and D were administered in a single dose of 200 mg/kg b.w. for cC and nC and 5 mg/kg b.w. for D. Association of cC or nC to D resulted in significant antinociceptive activity, and improved mechanical pressure stimulation and heat thresholds at 3, 5, 7 and 24 h (p < 0.03). The association of cC and nC with D (AIcC200D and AInC200D groups) showed significantly lower plasma and tissue levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) up to 2.5 times, with the best results in the group who received nC. Moreover, AInC200D presented the least severe histopathological changes with a reduced level of inflammation in the dermis and hypodermis. The combination of nC to D showed efficiency in reducing pain, inflammatory cytokines, and histological changes in acute inflammation.

Combining Multiple Biomarkers to Evaluate the Environmental Stress in Cyprinid Populations

The Vilariça River was restored in order to improve the fish habitat due to the construction of dams. However, due to the pressure that agriculture exerts on water resources, it is crucial to evaluate its consequences on fish health from a histological perspective. The gonad histopathological changes of two cyprinid species were assessed microscopically and evaluated with semi-quantitative analysis. Histopathological changes in the gonad, gill, and liver were used as biomarkers in the determination of the Integrated Biomarker Response (IBR), as well as to evaluate the histopathological scores between seasons. The observation of the gonad histopathological changes showed that interstitial fibrosis and macrophage aggregates were present exclusively in the Summer and more prevalent in males of large size. In addition, the histopathological scores of the three organs were higher in the Summer. In terms of the severity degree, 98% and 68% of the fish showed pronounced and severe histopathological changes in the gill and liver, respectively, while 28% of the fish showed slight histopathological changes in the gonad. The IBR showed higher values in the Summer and in the middle stream and downstream, which is in agreement with the metal concentrations. Thus, this work showed a relationship between the anomalies present in the fish organs and the quality of the water (classified as polluted). Therefore, minimization measures are presented, such as improving the irrigation methods, preserving the vegetation cover; promoting reforestation in degraded and burned areas; and improving the riparian vegetation.

Differential modulation of lung aquaporins among other pathophysiological markers in acute (Cl2 gas) and chronic (carbon nanoparticles, cigarette smoke) respiratory toxicity mouse models.

Inhaled toxic chemicals and particulates are known to disrupt lung homeostasis causing pulmonary toxicity and tissue injury. However, biomarkers of such exposures and their underlying mechanisms are poorly understood, especially for emerging toxicants such as engineered nanoparticles and chemical threat agents such as chlorine gas (Cl2). Aquaporins (AQPs), commonly referred to as water channels, are known to play roles in lung homeostasis and pathophysiology. However, little is known on their regulation in toxicant-induced lung injuries. Here, we compared four lung toxicity models namely, acute chemical exposure (Cl2)-, chronic particulate exposure (carbon nanotubes/CNT)-, chronic chemical exposure (cigarette smoke extract/CSE)-, and a chronic co-exposure (CNT + CSE)- model, for modulation of lung aquaporins (AQPs 1, 3, 4, and 5) in relation to other pathophysiological endpoints. These included markers of compromised state of lung mucosal lining [mucin 5b (MUC5B) and surfactant protein A (SP-A)] and lung-blood barrier [protein content in bronchoalveolar lavage (BAL) fluid and, cell tight junction proteins occludin and zona-occludens]. The results showed toxicity model-specific regulation of AQPs measured in terms of mRNA abundance. A differential upregulation was observed for AQP1 in acute Cl2 exposure model (14.71-fold; p = 0.002) and AQP3 in chronic CNT exposure model (3.83-fold; p = 0.044). In contrast, AQP4 was downregulated in chronic CSE model whereas AQP5 showed no significant change in any of the models. SP-A and MUC5B expression showed a decreasing pattern across all toxicity models except the acute Cl2 toxicity model, which showed a highly significant upregulation of MUC5B (25.95-fold; p = 0.003). This was consistent with other significant pathophysiological changes observed in this acute model, particularly a compromised lung epithelial-endothelial barrier indicated by significantly increased protein infiltration and expression of tight junction proteins, and more severe histopathological (structural and immunological) changes. To our knowledge, this is the first report on lung AQPs as molecular targets of the study toxicants. The differentially regulated AQPs, AQP1 in acute Cl2 exposure versus AQP3 in chronic CNT nanoparticle exposure, in conjunction with the corresponding differentially impacted pathophysiological endpoints (particularly MUC5B) could potentially serve as predictive markers of toxicant type-specific pulmonary injury and as candidates for future investigation for clinical intervention.

Toxicity of chronic waterborne zinc exposure in the hepatopancreas of white shrimp Litopenaeus vannamei

Zinc (Zn) is necessary for the survival of aquatic organisms; nevertheless, the accumulation of Zn in excessive amounts may have toxic consequences. Few studies focusing on the biochemical, morphological, and transcriptional effects of aqueous Zn in Litopenaeus vannamei have been reported, and the underlying toxic mechanism remains largely unknown. The present study was performed to investigate the growth performance, morphological alterations, physiological changes, and transcriptional responses after Zn exposure at 0 (control), 0.01, 0.1, and 1 mg/L concentrations for 30 days in white shrimp L. vannamei hepatopancreas. The results found that survival rate (SR) and growth performance were significantly reduced in 1 mg/L Zn group. Significant structural damage and significant Zn accumulation in hepatopancreas were observed. The activities of trypsin and amylase (AMS), and the total antioxidant capacity (T-AOC) were attenuated, while the production of reactive oxygen species (ROS) and malondialdehyde (MDA) content were significantly increased after Zn exposure. Many differentially expressed genes (DEGs) were obtained after Zn exposure, and the majority of these DEGs were downregulated. Ten DEGs involved in oxidative stress, immunological response, apoptosis, and other processes were selected for qRT-PCR validation and the expression profiles of these DEGs kept well consistent with the transcriptome data, which confirmed the accuracy and reliability of the transcriptome results. Subsequently, we screened 12 genes to examine the changes of expression in different concentrations in more detail. All the results implying that Zn exposure caused severe histopathological changes and increased Zn accumulation in hepatopancreas, altered immune, antioxidant and detoxifying response by regulating the gene expressions of related genes, and eventually might trigger apoptosis. These findings provide valuable information and a new perspective on the molecular toxicity of crustaceans in response to environmental heavy metal exposure.

H5N8 Subtype avian influenza virus isolated from migratory birds emerging in Eastern China possessed a high pathogenicity in mammals.

Novel H5N8 highly pathogenic avian influenza viruses (HPAIVs) bearing the clade 2.3.4.4b HA gene have been widely spread through wild migratory birds since 2020. One H5N8 HPAIV (A/Wild bird/Cixi/Cixi02/2020; here after Cixi02) was isolated from migratory birds in Zhejiang Province, Eastern China in 25 November 2020. However, its pathogenicity in avian and mammal remains unknown. Hemagglutinin gene genetic analysis indicated that Cixi02 virus belonged to the branch II of H5 clade 2.3.4.4b originated from Iraq in May 2020. Cixi02 virus showed a binding affinity to both SA α-2, 3-galactose (Gal) and SA α-2, 6 Gal receptors, good pH stability, thermostability, and replication ability in both avian and mammal cells. The poultry pathogenicity indicated that Cixi02 virus was lethal to chickens. Moreover, the mammalian pathogenicity showed that the 50% mouse lethal dose (MLD50 ) is 2.14 lgEID50 /50 μl, indicating a high pathogenicity in mice. Meanwhile, Cixi02 virus was widely detected in multiple organs, including heart, liver, spleen, lung, kidney, turbinate, and brain after nasal infection. In addition, we found high level gene expressions of TNF-α, IL-12p70, CXCL10, and IFN-α in lungs, IL-8 and IL-1β in brains, and observed severe histopathological change in lungs and brains. Collectedly, this study provided new insights on the pathogenic and zoonotic features of an H5N8 subtype AIV isolated from migratory birds.