Histomorphological Alterations Research Articles

Dietary supplementation of Lactobacillus brevis SAD ameliorates high-fat diet-induced hyperglycemia and associated metabolic issues in Swiss albino mice

ABSTRACT This study is aimed to investigate the therapeutic potential of a strain of Lactobacillus brevis (SAD) isolated from an ethnic fermented food consumed regularly by the native people of Darjeeling Himalayan region. Effects of regular consumption of this lactic acid bacteria (LAB) in ameliorating the pathophysiological conditions associated with hyperglycemia, insulin resistance, and obesity is studied in mice having induced obesity by high-fat diet (HFD). Key objectives included evaluating the strain’s impact on glycemic parameters, lipid profiles, colonic oxidative stress, inflammation, and gut integrity while assessing the metagenomic composition. For this study, male Swiss albino mice were grouped into Normal Diet (ND), HFD, and L. brevis SAD supplemented (LB) groups. It was found that, L. brevis SAD supplementation significantly improved glycemic parameters, insulin sensitivity, at the same time reduced glycation. It also mitigated obesity-related effects, such as weight gain, organ enlargement, and adverse lipid profiles. Moreover, the HFD group exhibited pronounced deterioration in gut integrity, evidenced by alterations in colonic histomorphology, concurrent with the development of obesity and hyperglycemia. In LB group gut integrity appeared to be restored in the histological study. Metagenomic analysis demonstrated the strain’s potential to modulate the gut microbial composition towards health promoting groups. This study demonstrates the therapeutic potential of L. brevis SAD in mitigating the adverse metabolic effects induced by HFD.

Histological evidence of hypothyroidism in mice chronically exposed to conventional farming

Worldwide, disorders of the thyroid gland are a growing concern; such can be caused by exposure to contaminants, including agrochemicals used in conventional agriculture, which act as endocrine disruptors. The purpose of this study is to evaluate whether or not exposure to an environment with conventional agriculture leads to thyroid disruption. Mus musculus were used as bioindicator species, captured in two sites: a farm where conventional agriculture is practiced, and a place without agriculture. Thyroid histomorphometric and morphologic data were analyzed. The impacts of the agricultural environment over the thyroid were revealed, as indications of hypothyroidism were observed in exposed mice: the area and volume of epithelial cells were much lower. Alterations in thyroid histomorphology were also observed: lower follicular sphericity, irregularly delimited epithelium and increased exfoliation into the colloid. These results highlight the need for transition from current conventional agricultural systems towards organic systems.

Histomorphological changes and alterations in the reproductive parameters of male wistar rats following exposure to ethanolic leaf extract of Datura stramonium (GEGEMU)

Histomorphological changes and alterations in the reproductive parameters of male wistar rats following exposure to ethanolic leaf extract of Datura stramonium (GEGEMU)

Study of the Histopathological Changes in the Heart in Postmortem/Autopsy Cases in Tertiary Care Center.

There has been a significant increase in global cardiac-related fatalities, particularly over the last five decades. In India, cardiovascular diseases have emerged as a prevalent condition. Moreover, the incidence of sudden cardiac death has shown a steady rise worldwide. The aim of this study was to research how frequently cardiac disease was found during postmortem/autopsy cases and how it affects sudden death. The research was conducted at the Department of Pathology, SRTR, GMC, Ambajogai, India. One hundred and eighty cardiac specimens were examined from October 2019 to October 2021. Formalin-fixed heart specimens had multiple sections cut out of them at 4-5 mm intervals. The tissues underwent preparation, followed by sectioning into 4 μm thickness using paraffin. The standard staining method of hematoxylin and eosin was used to stain the samples. Different cardiac histomorphological alterations were assessed. The study included 180 autopsy cases. The highest numbers of cases were observed in the age group of 31-40 years (20.2%). Triple-vessel involvement was seen in the majority of 61.5% of cases, out of 180 cases, 52.7% of cases were of hypertrophy. The research highlights the prevalence of cardiovascular diseases, particularly atherosclerosis and hypertrophy. Notably, sudden death cases showed a distinct distribution across the study years, with males being predominantly affected. This underscores the urgent need for preventive measures and interventions to prevalent cardiovascular conditions effectively.

Histomorphological Alterations in the Liver of Labeo rohita Exposed to Aflatoxin B1 and Quercetin-Enriched Diets

Histomorphological Alterations in the Liver of Labeo rohita Exposed to Aflatoxin B1 and Quercetin-Enriched Diets

Selenium and zinc supplementation mitigates metals-(loids) mixture- mediated cardiopulmonary toxicity via attenuation of antioxidant, anti-inflammatory and antiapoptotic mechanisms in female Sprague Dawley rats.

This study evaluated the cardiopulmonary protective effects of essential elements (Zn and Se) against heavy metals mixture (HMM) exposure. Twenty five female Sprague Dawley albino rats, divided in to five groups: controls were orally treated only with distilled water; next, group 2 was exposed to HMM with the following concentrations: 20mg/kg of Pb body weight, 0.40mg/kg of Hg, 0.56mg/kg of Mn, and 35mg/kg of Al. Groups 3, 4 and 5 were exposed to HMM and co-treated with zinc chloride (ZnCl2; 0.80mg/kg), sodium selenite (Na2SeO3;1.50mg/kg) and both zinc chloride and sodium selenite, respectively. The experiment lasted for 60days. Afterwards animals were sacrificed, and we conduced biochemical and histopathological examination of the heart and lungs. HMM only exposed animals had an increased levels of malondialdehyde (MDA) and nitric oxide (NO), increased IL-6 and TNF-α, attenuated SOD, GPx, CAT and GSH and caspase 3 in the heart and lungs. HMM affected NF-kB and Nrf2 in the heart muscle with histomorphological alterations. Zn and Se attenuated adverse effects of HMM exposure. Essential element supplementation ameliorated heavy metal cardiopulmonary intoxication in rats.

Cobalt exposure triggers impairments in cognitive and anxiety‐like behaviors, brain oxidative stress and inflammation, and hippocampo‐amygdala histomorphological alterations: Protective role of aqueous Prosopis africana seed extract

AbstractBackgroundCobalt toxicity has become a health concern in recent years, due to overexposure resulting in neurological impairments like dementia. With a growing interest in the therapeutic roles of herbs, in toxicity research, it’s worth looking into the curative effects of aqueous Prosopis africana seed extract, a plant rich in flavonoids on cobalt‐induced dementia‐like and anxiogenic behaviour.MethodWe treated rats with Cobalt (CoCl2) or CoCl2 in combination with aqueous Prosopis africana seed extract (PAE) orally for 14 days. Control rats received distilled water for the same period. Following treatments, behavioral experiments for cognition and anxiety, analysis for oxidative stress, inflammation, histological and immunohistochemical analysis were performed.ResultResults revealed that CoCl2 reduced the exploration time, recognition index in the novel object recognition test, percentage spontaneous alternation in the Y‐maze tests, and reduced open arm entry and duration in elevated plus‐maze. However, treatment with PAE improved these parameters to levels comparable with those of the control group. Furthermore, PAE therapy reduced CoCl2‐induced surge in hydrogen peroxide, malondialdehyde, TNF‐α and IL‐1β levels in brain homogenate, while also increasing superoxide dismutase and reduced reduced‐glutathione activities. CoCl2 exposure resulted in obvious features of neurodegeneration like nuclear disintegration, nuclear shrinkage, and cytoplasmic vacuolations of the cells of the hippocampus and amygdala, with an increased expression of GFAP. The hippocampal and amygdala histology improved after PAE administration, while exacerbated GFAP expressions were attenuated.ConclusionThese findings imply that PAE may be anxiolytic and can help reduce cognitive impairments and hippocampal damage caused by CoCl2 neurotoxicity, via mechanisms that involve attenuation of oxidative stress and neuroinflammation.

Pleiotropic attenuating effect of Ginkgo biloba against isoprenaline-induced myocardial infarction via improving Bcl-2/mTOR/ERK1/2/Na+, K+-ATPase activities

ObjectiveMyocardial infarction (MI) is linked to an imbalance in the supply and demand of blood oxygen in the heart muscles. Beta-blockers and calcium antagonists are just two of the common medications used to treat MI. However, these have reportedly been shown to be either ineffective or to have undesirable side effects. Extract of Ginkgo biloba leaves (GBE), a Chinese herbal product offers special compatibility benefits in therapeutic settings relating to inflammatory diseases and oxidative stress. In order to better understand how GBE affects MI in rats insulted by isoprenaline (ISO), the current study was designed. MethodsThe heart weight index, serum lipid profile, cardiac marker enzymes, endogenous antioxidants [catalase (CAT), superoxide dismutase (SOD), glutathione (GSH), nitrites and malondialdehyde (MDA)], inflammatory mediators [tumour necrosis factor alpha (TNF-α) and interleukin-6 (IL-6)], immunohistochemical expressions of B-cell lymphoma factor-2 (Bcl-2), extracellular signal-regulated kinase (ERK1/2), and mammalian target of rapamycin (mTOR) and histopathological analysis were used to assess the cardioprotective properties of GBE. ResultsThe findings showed that GBE effectively attenuated myocardial infarction by boosting the body’s natural antioxidant defense system and reducing the release of inflammatory cytokines as well as heart injury marker enzymes. The expression of Bcl-2, ERK1/2 and mTOR was increased while the histomorphological alterations were reversed. ConclusionThe cardioprotective effects of GBE may be due to a mechanism involving increased Bcl-2/mTOR/ERK1/2/Na+, K+-ATPase activity.

The anti-liver fibrosis effect of Tibetan medicine (Qiwei Tiexie capsule) is related to the inhibition of NLRP3 inflammasome activation in vivo and in vitro

Ethnopharmacological relevanceQiwei Tiexie capsule (QWTX) is an improved form of a classical prescription of Tibetan medicine—Qiwei Tiexie pill. It has been employed in the treatment of a variety of chronic liver disorders, including liver fibrosis. Uncertainty still exists regarding the mechanism of QWTX action in liver fibrosis. Aim of the studyConfirm the anti-liver fibrosis effect of QWTX and reveal its mechanism from the perspective of NOD-like receptor protein 3 (NLRP3) inflammasome activation. Materials and methodsIn vivo experiment: A rat model of carbon tetrachloride -induced liver fibrosis was constructed. All rats were randomly divided into six groups: a control group, a model group, a group receiving the positive drug (Biejia Ruangan tablet), and three groups receiving QWTX at high, medium, and low doses. The contents of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and total bilirubin (TBil) were detected in serum. Hematoxylin and eosin staining and Masson's staining were used to assess the histomorphological alteration of the liver. The levels of glutathione peroxidase, hydroxyproline, tumor necrosis factor alpha (TNF-α), and interleukin 1 beta (IL-1β) in the liver were determined using the corresponding detection kits. Real-time polymerase chain reaction, immunofluorescence, and western blotting were used to determine the expression levels of NLRP3, adaptor protein (ASC), caspase-1, and alpha-smooth muscle actin (α-SMA). In vitro experiment: Four groups of rat hepatic stellate cell line (HSC-T6) cells were created: the control group, the low-dose QWTX group (0.05 mg/mL), the medium-dose QWTX group (0.1 mg/mL), and the high-dose QWTX group (0.2 mg/mL). Cell viability was assessed using a cell counting kit, and the amounts of collagen type I (Col I) and IL-1β in the cell lysate were measured using an enzyme-linked immunosorbent assay kit. The mRNA and protein expression of NLRP3, ASC, caspase-1, and α-SMA were also estimated. ResultsQWTX had an inhibitory effect on liver fibrosis and a negative effect on HSC activation, while it improved liver histopathological injury and abnormal liver function and increased hydroxyproline content and glutathione peroxidase activity in vivo. QWTX decreased the expression of α-SMA, NLRP3, caspase-1, ASC, and IL-1β both in vitro and in vivo. ConclusionsTibetan medicine QWTX had a significant anti-liver fibrosis effect that was related to the inhibition of NLRP3 inflammasome activation in vivo and in vitro.

Dose-Response Relationship and Histo-morphological Alterations on Oreochromis niloticus Juveniles Following Exposure to Ethanolic Extract of Latana camara

Aim: This research intends to evaluate the toxicity of latana camara and ascertain if it can be useful as a plant-based additive in the formulation of fish feed.
 Study Design: The study was conducted using two replicate (Batch A and B) for 96 hours under controlled laboratory conditions. Five concentrations ranging from 0, 5, 10, 15, and 20 mg/l were prepared from the ethanolic extract (EE) of L. camara for the toxicity test.
 Place and Duration of Study: Experiment was conducted in Akwa Ibom State University, Obio Akpa Campus, Akwa Ibom State between February, 2023 and June, 2023.
 Methodology: Total of two hundred (200) juveniles were collected for the studies. One hundred (100) juveniles were used during range findings test and the left over for the actual toxicity test. Prior to commencement of the toxicity test, experimental fish were allowed to acclimatize. Each of the ten (10) plastic aquaria was stocked with ten (10) O. niloticus juveniles. The prepared plant extract at varying concentration was added to each stock aquaria and allowed to stand for 96 hours for mortality examination. Gills tissues were isolated from the fish samples in each of the concentration after 96 hours for histopathological examinations.
 Results: The experimental animals showed differential percentage mortalities with toxicant concentrations. The 96 hours LC50 for O. niloticus for both batches (A and B) was given at 7.346 mg/l representing a log transformed concentration of 0.866 mg/l. The different batches of O. niloticus (P = .05) had no significant difference in mortality. The results of the present study suggest that the EE of L. camara had severe impacts on the test organism resulting in mortality. The effects of L. camara on the gills of O. niloticus Juveniles showed severe impacts on the test organisms. Five samples were taken from each of the concentrations to examine the effects of the extract on the gills of O. niloticus. There were no observed changes in the gills of the control group as compared to other treatment which showed evidence of histological alterations.
 Conclusion: From the findings, it is observed that extract obtained from L. camara is toxic to aquatic life and cannot be recommended as a plant-based additive in the formulation of fish feed.

Effects of repeatedly heated cooking oil consumption in mice: a study on health implications

Background: Cooking oils are a major part of human diets, but repeated use of heated oils can have detrimental effects on consumer health. This study aims to investigate the impact of different heating grades of vegetable oils on the hemato-biochemical parameters and vital organs like the heart, liver, kidney, and intestine in mice. Methods: Thirty mice were randomly assigned to different treatment groups, including a control group (diet only), unheated cooking oil (UHCO) group, single heated cooking oil (SHCO) group, three times repeatedly heated cooking oil (3RHCO) group, and repeatedly heated cooking oil (ReHCO) group. Blood and organ samples were collected on day 31 to investigate hemato-biochemical parameters and histo-morphological alterations in response to the oil treatments. Results: The oil-treated groups showed significant (P<0.05) decreases in the total erythrocyte, leukocyte, and hemoglobin levels. Meanwhile, serum levels of total cholesterol, triglyceride, high-density lipoprotein, glucose, and creatinine increased significantly (P<0.05), while low-density lipoprotein and protein levels dropped markedly in the treatment groups. Severe histo-morphological alterations were also found in the liver (hepatocytic degeneration with hydropic change in the 3RHCO and ReHCO groups), kidney (glomerular atrophy with increased glomerular space, tubular degeneration, and lymphocytic infiltration in the SHCO, 3RHCO, and ReHCO groups), and colon (lymphocytic infiltration in the mucosal layer of ReHCO group). Conclusions: These findings suggest that the consumption of heated oils can have severe adverse effects on consumers' health, leading to alterations in blood chemistry and damage to vital organs.

Evidence of enteritis, hepatic steatosis and jaundice in juvenile pirarucu (Arapaima gigas) fed high levels of soybean meal

ABSTRACT This study evaluated the effects of replacing dietary fishmeal with increasing levels of soybean meal (0, 15, 30, 45 and 60% corresponding to 0, 11, 22, 33 and 44% inclusion in the formulated diet) on the intestine and liver histomorphology of juvenile pirarucu Arapaima gigas (233 ± 12 g; mean ± SD) fed for 120 days. At the end of the feeding trial, fish fed diets with higher soybean levels (45 and 60%) presented clinical signs such as appetite loss, cachexia and jaundice. In addition, fish fed on 45% and 60% soybean meal had histomorphological alterations in the middle and posterior portions of the intestinal mucosa (enteritis) and high liver lipid deposition (steatosis). Therefore, 45% or 60% of soybean meal is not recommended; however, a replacement of fishmeal by soybean meal of up to 30% (22% inclusion) can be used in diets to feed juvenile pirarucu for long periods.

Feeding deterrence and midgut histomorphological alterations in Spodoptera littoralis larvae treated with separate and combined imidacloprid and spinosad

Feeding deterrence and midgut histomorphological alterations in Spodoptera littoralis larvae treated with separate and combined imidacloprid and spinosad

Alterations in histomorphology, biochemical parameters and gross morphometry in liver of albino rats following administration of rifampicin and isoniazid

Background: Rifampicin (RIF) and Isoniazid (INH) are two main medicinal drugs used as first line regimen in the treatment of Tuberculosis. These drugs have shown to induce hepatotoxicity and alteration of liver parenchyma upon their administration. The aim of this study was to investigate the gross morphometric, histomorphological and biochemical effect of INH and RIF on the liver architecture. Methods: Adult albino rats with an average weight of between 150g to 250g were used in the study. A total of 12 rats were randomly assigned into 2 groups each group containing 6 rats. Group one was the control group (no intervention). Group two was the experimental group, given INH (50mg/kgbwt) and RIF (50mg/kgbwt). At the end of the experiment, after 21 days, the albino rats were sacrificed humanely, liver harvested, weighed and the gross parameters measured using a ruler and caliper. Blood samples were taken for liver serum biomarkers analysis. Thereafter, the livers were processed and stained with Haematoxylin and Eosin for histological examination. Results: There was a significant (p value < 0.0001) decrease in gross morphometric measurements of the weight, length, width, thickness in experimental group. The three selected liver biochemical parameters (ALT, AST and ALP) were observed to be high above their normal ranges. The liver sections in positive control group showed deranged histomorphological features. Conclusion: co-administration of Rifampicin and Isoniazid can have effect on gross morphometric, histomorphology and liver biochemical parameters of the liver therefore, their administration should be done in caution.

Alzheimer's Disease-like Pathological Features in the Dorsal Hippocampus of Wild-Type Rats Subjected to Methionine-Diet-Evoked Mild Hyperhomocysteinaemia.

Multifactorial interactions, including nutritional state, likely participate in neurodegeneration's pathogenesis and evolution. Dysregulation in methionine (Met) metabolism could lead to the development of hyperhomocysteinaemia (hHcy), playing an important role in neuronal dysfunction, which could potentially lead to the development of Alzheimer's disease (AD)-like pathological features. This study combines proton magnetic resonance spectroscopy (1H MRS) with immunohistochemical analysis to examine changes in the metabolic ratio and histomorphological alterations in the dorsal rat hippocampus (dentate gyrus-DG) subjected to a high Met diet. Male Wistar rats (420-480 g) underwent hHcy evoked by a Met-enriched diet (2 g/kg of weight/day) lasting four weeks. Changes in the metabolic ratio profile and significant histomorphological alterations have been found in the DG of hHcy rats. We have detected increased morphologically changed neurons and glial cells with increased neurogenic markers and apolipoprotein E positivity parallel with a diminished immunosignal for the N-Methyl-D-Aspartate receptor 1 in hHcy animals. A Met diet induced hHcy, likely via direct Hcy neurotoxicity, an interference with one carbon unit metabolism, and/or epigenetic regulation. These conditions lead to the progression of neurodegeneration and the promotion of AD-like pathological features in the less vulnerable hippocampal DG, which presents a plausible therapeutic target.

Toxicology of arsenate, arsenite, cadmium, lead, chromium, and nickel in testes of adult Swiss mice after chronic exposure by intraperitoneal route

BackgroundSome residues such as the heavy metals cadmium (Cd), lead (Pb), chromium (Cr VI), nickel (Ni), and arsenic (As), this last one in its oxidized forms + 5 (arsenate) and + 3 (arsenite), can cause injuries to human health, so they are currently considered environmental health emergencies. In the testis, heavy metals can cause morphological and functional damage due to constant exposure acting chronically in individuals. Thus, we aimed to determine the toxicological mechanism of As+5, As+3, Cd, Cr VI, and Ni that leads to testicular damage and establish for the first time an order of toxicity among these studied heavy metals. MethodsForty-two Swiss mice at reproductive age (140 days) were used, randomly distributed into seven experimental groups (n = 6). Exposure to heavy metals was weekly performed, by intraperitoneal route. Group 1 received 0.7 mL 0.9% saline (control), and the other groups received 1.5 mg/ kg of As+5, As+3, Cd, Pb, Cr VI, or Ni, for six weeks. ResultsThese studied heavy metals did not accumulate in the testis tissue. However, exposure to Ni induced moderate pathologies in the seminiferous tubules, plus changes in the tunica propria, blood vessels, lymphatic space, and carbonyl protein levels. Cd exposure caused moderate tubular histopathologies and changes in the blood vessels and lymphatic space. Cr VI induced slight tubular histopathologies, changes in the lymphatic space, blood vessels, and SOD activity. Pb and As+3 exposure triggered moderate tubular pathologies and changes in the SOD activity and carbonyl protein levels, respectively. Finally, As+5 induced only slight tubular pathologies. ConclusionThe testicular histopathologies caused by the studied heavy metals are mainly triggered by changes in testicular oxidative balance. Based on our findings of histomorphological alterations, the toxicity order among the heavy metals is Ni>Cd>Cr(VI)>PbAs+3 >As+5. However, considering oxidative stress results, we propose the following testicular toxicity order for these heavy metals: Ni>As+3 > Cd>Cr(VI)>Pb>As+5. Ni exposure shows the most harmful among the heavy metals to the testis.

Mammary gland development in male rats perinatally exposed to propiconazole, glyphosate, or their mixture

This study aimed to assess whether perinatal exposure to propiconazole (PRO), glyphosate (GLY) or their mixture (PROGLY) alters key endocrine pathways and the development of the male rat mammary gland. To this end, pregnant rats were orally exposed to vehicle, PRO, GLY, or a mixture of PRO and GLY from gestation day 9 until weaning. Male offspring were euthanized on postnatal day (PND) 21 and PND60. On PND21, GLY-exposed rats showed reduced mammary epithelial cell proliferation, whereas PRO-exposed ones showed increased ductal p-Erk1/2 expression without histomorphological alterations. On PND60, GLY-exposed rats showed reduced mammary gland area and estrogen receptor alpha expression and increased aromatase expression, whereas PRO-exposed ones showed enhanced lobuloalveolar development and increased lobular hyperplasia. However, PROGLY did not modify any of the endpoints evaluated. In summary, PRO and GLY modified the expression of key molecules and the development of the male mammary gland individually but not together.

PHYTOCHEMICALS AND TOXICOLOGICAL EFFECTS OF HERBAL DRINK (HD) ON LIVER AND KIDNEY IN WISTAR RATS

Herbal medicine use is on the increase worldwide and many studies have associated their use with hepatotoxicity and nephrotoxicity. Herbal drink (HD) is a commercial herbal supplement mainly used in Uganda. In this study, we evaluated the phytochemical composition, and acute and sub-chronic toxicological effects of HD on hepatorenal integrity in Wistar rats. Modified Lorke’s method was employed in the acute toxicity study. For the sub-chronic toxicity study, a total of sixteen (N= 16) Wistar rats were randomly allotted to four groups of four rats each (n=4). Normal control group, NC (normal dose equivalent of distilled water), Group T1 (half normal dose of HD), Group T2 (normally recommended dose equivalent of HD),and Group T3 (twice normal recommended dose of HD). All treatments were done by intragastric gavage for a period of twentyeight days. Phytochemical analysis of HD showed the presence of alkaloids, glycosides, tannins, phenolic compounds, flavonoids, saponins, and carbohydrates. The animals were able to tolerate a dose of up to 5000 mg/Kg. It was demonstratedthat continuous twenty-eight-day treatment with HD does not significantly (p>0.0 alter kidney and liver function parametersin Wistar rats. This observation concurs with the histopathological findings of the liver and kidney tissues which showed no remarkable histomorphological alterations. Regular intake of HD does not have any significant toxic effects on the liver and kidney in Wistar rats.

Structural and proteomic repercussions of growth hormone receptor deficiency on the pituitary gland: Lessons from a translational pig model.

Growth hormone receptor deficiency (GHRD) results in low serum insulin-like growth factor 1 (IGF1) and high, but non-functional serum growth hormone (GH) levels in human Laron syndrome (LS) patients and animal models. This study investigated the quantitative histomorphological and molecular alterations associated with GHRD. Pituitary glands from 6 months old growth hormone receptor deficient (GHR-KO) and control pigs were analyzed using a quantitative histomorphological approach in paraffin (9 GHR-KO [5 males, 4 females] vs. 11 controls [5 males, 6 females]), ultrathin sections tissue sections (3 male GHR-KO vs. 3 male controls) and label-free proteomics (4 GHR-KO vs. 4 control pigs [2 per sex]). GHR-KO pigs displayed reduced body weights (60% reduction in comparison to controls; p < .0001) and decreased pituitary volumes (54% reduction in comparison to controls; p < .0001). The volume proportion of the adenohypophysis did not differ in GHR-KO and control pituitaries (65% vs. 71%; p = .0506) and GHR-KO adenohypophyses displayed a reduced absolute volume but an unaltered volume density of somatotrophs in comparison to controls (21% vs. 18%; p = .3164). In GHR-KO pigs, somatotroph cells displayed a significantly reduced volume density of granules (23.5%) as compared to controls (67.7%; p < .0001). Holistic proteome analysis of adenohypophysis samples identified 4660 proteins, of which 592 were differentially abundant between the GHR-KO and control groups. In GHR-KO samples, the abundance of somatotropin precursor was decreased, whereas increased abundances of proteins involved in protein production, transport and endoplasmic reticulum (ER) stress were revealed. Increased protein production and secretion as well as significantly reduced proportion of GH-storing granules in somatotroph cells of the adenohypophysis without an increase in volume density of somatotroph cells in the adenohypophysis could explain elevated serum GH levels in GHR-KO pigs.

Detection of Diclofenac-Induced Alterations in Rainbow Trout (Oncorhynchus mykiss) Using Quantitative Stereological Methods.

In 2013, the nonsteroidal anti-inflammatory drug diclofenac (DCF) was included in the watch list for emerging pollutants under the European Union Water Framework Directive. Frequently, monitoring data revealed DCF concentrations in surface waters exceeding the proposed environmental quality standards of 0.04 µg L-1 and 0.126 µg L-1 . In recent literature, the possible effects of DCF on fish are discussed controversially. To contribute to a realistic risk assessment of DCF, a 28-day exposure experiment was carried on rainbow trout (Oncorhynchus mykiss). To warrant reliability of data, experiments were conducted considering the Criteria for Reporting and Evaluating Ecotoxicity Data. The test concentrations of DCF used (0.1, 0.5, 1, 5, 25, and 100 µg L-1 ) also included environmentally relevant concentrations. The lowest-observed-effect concentration (LOEC) for a significant decrease in the plasma concentrations of the DCF biomarker prostaglandin E2 was 0.5 µg L-1 (male fish). For objective evaluation of relevant histomorphological parameters of gills and trunk kidneys, unbiased quantitative stereological methods were applied. In the gills, significant increases in the thickness of the secondary lamella and in the true harmonic mean of barrier thickness in secondary lamellae were present at DCF concentrations of 25 µg L-1 and 100 µg L-1 . In the trunk kidneys, the absolute and relative volumes of nephrons were significantly decreased, paralleled by a significant increase of the volume of the interstitial renal tissue. With regard to quantitative histomorphological alterations in the trunk kidney, the observed LOEC was 0.5 µg L-1 . The quantitative histomorphological analyses that were conducted allow identification and objective quantification of even subtle but significant morphological effects and thus provide an important contribution for the comparability of study results for the determination of no-observed-effect concentrations(NOEC). Environ Toxicol Chem 2023;42:859-872. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.