Histopathological Alterations Research Articles

Histopathology based study of Nile tilapia fish (Oreochromis niloticus) as a biomarker for water pollution evaluation in the southern gulf of Lake Tana, Ethiopia

In the past decade, the increasing distribution of pollutants in the aquatic environment has been observed, causing integrative effects on fish. Likewise, due to anthropogenic activities, the southern gulf of Lake Tana is an impacted region, and the production of Nile tilapia fish is reduced. For this reason, the aim of this study was to conduct a histopathological-based study of 48 Nile tilapia fishes’ health status at the southern gulf of Lake Tana and aquaculture using a cross-sectional study from February 2023 to May 2023. The study evaluated the histopathology of the gill, liver, gonads, and spleen organs using descriptive statistics accompanied by a 2 × 2 contingency table and t-test analysis. During the study, different histological alterations were detected, and the numbers of fish affected by a specific histological alteration were presented as percentage prevalence; hence, from the total fish examined, hyperplasia (54.15%), followed by pigment deposits (52%), hemorrhage (50%), and immune cell infiltration (50%), respectively, were the most frequently detected alterations. However, Nile tilapias from the southern gulf of Lake Tana were 1.4 (odds ratio) times more likely to show histopathological alterations than those from aquaculture, although statistically, was not significant (p > 0.05). In addition, the study found the mean value of the fish index (95.3) and regressive indices of the gill (13.6), liver (14.8), and gonad (12.3); moreover, the inflammatory indices of the spleen organ (11.3) and mean severity grade value of the gill (2.35) and gonad (1.7) organs, respectively, were obtained from the southern gulf of Lake Tana, and all those values were significantly higher (p < 0.05) from this site as compared to the aquaculture. In general, it has been found that tilapias from the southern gulf of Lake Tana showed higher pathological severity as compared with aquaculture. Among the four target organs evaluated, liver organs were observed to be the most damaged, while gonads were the least impacted organs. Therefore, it has been concluded that tilapia fish are living in abnormal conditions, so to ensure a sustainable fishery, water pollutant sources from Bahirdar city must receive proper attention, and future studies should consider age differences, seasonal variation, and the detection of specific pollutants.

Impact of liposomal hesperetin in broilers: prospects for improving performance, antioxidant potential, immunity, and resistance against Listeria monocytogenes

ABSTRACT Liposomal encapsulated phytogenics, such as liposomal hesperetin, are considered novel substitutes for antibiotics in the broiler industry owing to their improved nutritional and therapeutic properties. Therefore, our key goal was to investigate liposomal hesperetin impact on broiler growth performance, health, antioxidant status, tight junction proteins (TJP), and resistance against Listeria monocytogenes. Four broiler groups were fed 0, 150, 250, or 400 mg/kg of liposomal hesperetin-supplemented diets and experimentally infected with L. monocytogenes strain. Herein, liposomal hesperetin, especially at higher concentrations, augmented broilers FCR with upregulation of genes encoding TJP (occludin, JAM-2, MUC-2), and antioxidant attributes (GPX-1, SOD-1, CAT, HO-1, NQO1, COX2), which reflect enhancing health and welfare of broilers. Muscle antioxidant biomarkers were enhanced; meanwhile, muscle MDA, ROS, and H2O2 levels were reduced in response to 400 mg/kg of liposomal hesperetin. Liposomal hesperetin fortification reduced L. monocytogenes loads and expression levels of its virulence-related genes (flaA, hlyA, and ami). Remarkably, histopathological alterations in intestinal and brain tissues of L. monocytogenes-infected broilers were restored post-inclusion at higher levels of liposomal hesperetin, which reflects increase of the birds’ resistance to L. monocytogenes infection. Transcription levels of genes encoding cytokines/chemokines (MyD88, AVBD6, CCL20, IL-1β, IL-18), and autophagy (Bcl-2, LC3, AMPK, AKT, CHOP, Bip, p62, XBP1) were ameliorated following dietary liposomal hesperetin fortification, which suggests enhancement of the birds’ immunity and health. Collectively, our research recommends liposomal hesperetin application in broiler diets owing to its promoting impact on growth performance, antioxidant status, immunity, health, and welfare besides its antibacterial, and antivirulence characteristics to fight against L. monocytogenes.

Fusarium begoniae metabolites: a promising larvicidal, pupicidal potential, histopathological alterations and detoxifications enzyme profiles of medically important mosquito vector Aedes aegypti, Culex quinquefasciatus and Anopheles stephensi.

The online version contains supplementary material available at 10.1007/s13205-024-04061-z.

Hepatic and immune modulatory effectiveness of lactoferrin loaded Selenium nanoparticles on bleomycin induced hepatic injury

This study aimed to estimate the hepatic and immune ameliorating potential of extracted bovine lactoferrin (LF), Selenium nanoparticles (SeNPs) or their combination (LF/SeNPs) against bleomycin (BLM) induced hepatic injury. Fifty adult male rats (160–200 g) were equally divided into five groups: (1) the saline control group, (2) BLM-injected (15 mg/kg twice a week, ip), and (3–5) groups treated orally with LF (200 mg/kg/day), SeNPs (0.0486 mg/kg/day) or LF/SeNPs combination (200.0486 mg/kg/day) for 6 weeks post BLM-intoxication. Blood and liver samples were subjected to biochemical, histopathological, and immunohistochemical analyses. The results revealed that BLM caused a significant increase in hepatic lipid peroxidation and nitric oxide, as well as serum markers of liver functions (AST, ALT and GGT activities), and levels of GM-CSF, CD4, TNF-α, IL-1β, TGF-β1, fibronectin, triglycerides, cholesterol and LDL-C. Additionally, hepatic glutathione, Na+/K+-ATPase, and glutathione peroxidase, as well as serum HDL-C, total protein and albumin levels were significantly reduced. Moreover, BLM injection resulted in marked histopathological alterations and severe expression of caspase 3. Post-treatment of BLM-intoxicated rats with LF, SeNPs or LF/SeNPs combination obviously improved the BLM-induced hepatic damages; this was achieved from the marked modulations in the mentioned parameters, besides improving the histopathological hepatic architecture. It is worth mentioning that LF/SeNPs exerted the greatest potency. In conclusion, the obtained results demonstrated that LF, SeNPs and LF/SeNPs succeeded in attenuating the BLM-induced hepatic dysfunction. Therefore, these supplements might be used to protect against drug-associated side effects.

Potential Antiallergic Activity of Two Chemically/Enzymatically-Modified Natural Products Against Active Atopic and Systemic Anaphylaxes in CD1 Mice Models

ABSTRACT Introduction Anaphylaxis is a globally increasing allergic reaction that is often fatal. Recently, our previous study reported the possibility of using the modified natural products “sodium R-lipoate (NaRLA) and enzymatically modified isoquercitrin (EMIQ)“ as potential novel safe agents against the non-immunological-degranulation of mast cells. Methods Here, we extended our previous findings by determining the antianaphylactic activity of 50 and 100 mg/kg body weight of NaRLA and EMIQ (given orally and prior to local or systemic challenge) in mice models of ovalbumin (OVA)-induced IgE-dependent active cutaneous anaphylaxis (ACA) and active systemic anaphylaxis (ASA) in comparison with sulfasalazine (SSZ, amast cell stabilizer). Results The pre-treatment of mice with NaRLA or EMIQ completely succeeded, as SSZ, in suppression of the increased vascular permeability associated with IgE-dependent ACA and protected the OVA-sensitized mice from fatal ASA by reducing (p < .001) the skin mast cell degranulation, the elevated peritoneal histamine and interleukin-4 levels, along with decreasing the associated sever gastrointestinal and lung histopathological alterations and inflammation. The high dose of EMIQ prevented death in 70% of mice with anaphylactic shock, better than SSZ. Discussion Our data indicated that NaRLA and EMIQ may be potential prophylactic and therapeutic candidates for the alleviation of atopic and systemic anaphylaxis.

Supplementing Nile tilapia (Oreochromis niloticus) (Linnaeus, 1758) larvae with dietary beta-glucan could improve their growth, survival, immune function, intestinal and liver histomorphology

Abstract Β-glucan exerts a positive impact and consider as a functional feed additives to enhance fish growth, immunity, control disease outbreaks and pathogen infections by increasing functional, immune and decreasing harmful responses. The present study aimed to determine the most effective dietary level of beta-glucan (βG) to improve Nile tilapia Oreochromis niloticus (L.) fry growth performance, feed utilization, and hematological indicators. Seven isonitrogenous (32% crude protein) and isocaloric diets (18 MJ/kg) were formulated. All diets were identical except for the variation in βG levels.The basal experimental diet (control diet) had no βG added. Diets 2–7 each contained βG at levels of 250, 500, 1000, 2000, 4000, and 8000 mg/kg diet, respectively. A total of 840 fish with an average body weight of 0.2±0.01 g were allocated into the seven experimental treatments (in triplicate). The trial lasted for 120 days, Nile tilapia fry growth performance and feed utilization were significantly (P≤0.05) higher in all treatments receiving βG diets than in the control diet. The fish fed with ßG0.1 diet showed the highest significant growth induces and the best FCR values. The survival (S%) of the fish also showed a significant increase in the βG diets up to ßG0.8 levels, when compared to the other experimental diet groups. All hematological parameters increased (P≤0.05) in fish fed with a diet supplemented up to ßG0.8 compared to the control group. Histological examination of the intestine and liver in the control group revealed histopathological alterations in the villi. On the other hand, the groups treated with βG had longer and structurally normal villi. The most well-preserved intestinal tissue and the tallest villi were observed in ßG0.1, followed by ßG0.050 and ßG0.8 groups. In the liver, the control group exhibited fatty degeneration, necrosis and pyknosis whereas the ßG0.1 group displayed the most preserved hepatic tissue, followed by ßG0.8 and ßG0.2 groups. The results indicate that according to FBW-based broken-line model analysis, the optimal dietary level of βG for Nile tilapia fry to exhibit superior growth and diet utilization efficiency associated with the best FCR for Nile tilapia should be 0.12% βG /kg diet. To increase the immunity of fish and improve the properties of hematological and histopathological indices, the dose can be increased to 0.8% βG without any adverse effects.

Plasma activated water effects on behavior, performance, carcass quality, biochemical changes, and histopathological alterations in quail

BackgroundPlasma-activated water (PAW) is an innovative promising technology which could be applied to improve poultry health. The current study investigated the effects of drinking water supply with PAW on quail behaviour, performance, biochemical parameters, carcass quality, intestinal microbial populations, and internal organs histopathology. A total of 54 twenty-one-day-old Japanese quail chicks were randomly allotted to three treatments provided with PAW at doses 0, 1 ml (PAW-1), and 2 ml (PAW-2) per one litter drinking water. Each treatment contained 6 replicates (3 birds/ cage; one male and two females).ResultsThe results clarified that there were no significant (P > 0.05) changes in behaviour, and performance. For the biochemical indicators, the PAW-1 group showed significantly higher serum H2O2, total protein and globulin levels compared with the other groups (P = 0.015, < 0.001, and 0.019; respectively). PAW groups had significantly lower serum creatinine and urea levels than the control (P = 0.003). For the carcass quality, the internal organs relative weight between different treatments was not changed. In contrast, there was a significant increase in the meat colour, taste, and overall acceptance scores in PAW groups compared with the control one (P = 0.013, 0.001, and < 0.001; respectively). For the intestinal microbial population, lactobacilli count was significantly higher in PAW-2 compared with the control group (P = 0.014), while there were no changes in the total bacterial count between different treatment groups. Moreover, mild histological changes were recorded in the intestine, liver, and spleen of PAW groups especially PAW-2 compared with the control one.ConclusionsPAW offered benefits, such as reducing creatine and urea levels, improving meat characteristics, and increasing lactobacilli count, all of which are crucial for sustainable quail farming. Therefore, further research is needed.

Umbelliprenin attenuates comorbid behavioral disorders in acetic acid-induced colitis in mice: mechanistic insights into hippocampal oxidative stress and neuroinflammation.

Inflammatory bowel disease (IBD) is often accompanied by psychiatric disorders. Emerging evidence suggests that neuroinflammation and oxidative stress contribute to the psychiatric symptoms associated with IBD. Umbelliprenin (UMB) possesses several pharmacological properties, including anti-inflammatory and antioxidant effects. This study aimed to investigate the protective effects of UMB on comorbid behavioral disorders in a mouse model of experimental colitis, focusing on its potential anti-neuroinflammatory and antioxidant activities. After inducing colitis with acetic acid, male NMRI mice were treated for 7 consecutive days with UMB, saline, or dexamethasone. Behavioral assessments included the forced swimming test (FST), splash test, open field test (OFT), and elevated plus maze (EPM). Histopathological changes in the colon were evaluated, and total antioxidant capacity (TAC), malondialdehyde (MDA) levels, and the expression of inflammatory genes (TNFα, IL1β, and TLR4) were measured in the hippocampus. Colitis was associated with increased immobility time in the FST, reduced entries and time spent in the open arms of the EPM, decreased grooming behavior in the splash test, and reduced time spent in the central zone of the OFT. Colitis also resulted in a reduction in TAC and an increase in MDA levels and inflammatory gene expression in the hippocampus. UMB treatment mitigated the behavioral disorders associated with colitis, reduced neuroinflammation and oxidative stress in the hippocampus, and alleviated histopathological alterations in the colon. In conclusion, UMB may reduce behavioral disorders induced by colitis by decreasing oxidative stress and neuroinflammation in the hippocampus.

Zingiber officinale Ameliorates Acute Toxoplasmosis-Induced Pathology in Mice.

Toxoplasma gondii (T. gondii) infects one third of the world's population with significant illness, mainly among immunocompromised individuals and pregnant women. Treatment options for toxoplasmosis are limited which signifies the need for novel, potent, and safe therapeutic options. The goal of this study was to assess the effectiveness of the ethanolic extract of Zingiber officinale (Z. officinale) in treating mice infected with the RH T. gondii strain. Gas Chromatography/Mass Spectrometry (GC/MS) was used to identify components of ethanolic extract of Z. officinale. A total of 80 mice were randomly allocated into four experimental groups that contained 20 mice each. The first group was left uninfected (uninfected control), while three groups were infected with T. gondii RH virulent strain tachyzoites at 2500 tachyzoites/mouse. One infected group was left untreated (infected, untreated), whereas the other two groups were treated orally with either spiramycin (positive control) or Z. officinale ethanolic extract at doses of 200mg/kg and 500mg/kg, respectively for 5 days, starting the day of infection. Ten mice from each group were used to assess mice survival in different groups, whereas the other ten mice in each group were sacrificed on the 5th day post-infectin (dpi) to estimate the treatment efficacy by quantifying liver parasite load, liver function, nitric oxide (NO) production, and levels of antioxidant enzymes. Additionally, histopathological studies were performed to evaluate the therapeutic effect of Z. officinale treatment on toxoplasmosis-induced pathological alterations in liver, brain, and spleen. Treatment with Z. officinale ethanolic extract extended the survival of mice till 9th dpi compared to 7th dpi in infected untreated mice. Higher percentage of mice survived in Z. officinale-treated group compared to spiramycin-treatment group at different time points. Liver parasite loads were significantly lower in Z. officinale extract-treated mice and spiramycin-treated mice compared to infected untreated mice which correlated with significantly lower levels of serum liver enzymes (ALT, AST) and nitric oxide (NO), as well as significantly higher catalase (CAT) antioxidant enzyme activity. Scanning electron microscopy (SEM) examination of tachyzoites from the peritoneal fluid revealed marked damage in tachyzoites from Z. officinale-treated group compared to that from infected untreated mice. Moreover, treatment with Z. officinale ethanolic extract alleviated infection-induced pathological alterations and restored normal tissue morphology of liver, brain, and spleen. Our results demonstrated that Z. officinale treatment reduced parasite burden and reversed histopathological and biochemical alterations in acute murine toxoplasmosis. These findings support the potential utility of Z. officinale as a future effective natural therapeutic for toxoplasmosis. Further studies are needed to determine the effective active ingredient in Z. officinale extract that can be further optimized for treatment of toxoplasmosis.

Antagonistic effects of Bacillus subtilis-derived chitosan nanoparticles on growth performance, stress biomarkers, and histological alterations of cadmium-intoxicated Nile tilapia fingerlings

Heavy metals including cadmium (Cd) are one of the major persistent and non-biodegradable wastewater pollutants. However, Cd reaches the aquatic ecosystem via industrial and agricultural waste discharges and causes serious deterioration in the welfare status of aquatic animals. The use of feed supplements with immune-stimulants to mitigate the toxic influences of heavy metals including Cd is a much more intriguing point. Thus, the current experiment used the bio-synthetized chitosan nanoparticles derived from Bacillus subtilis (Bs-CNPs) as a feed supplement and evaluated its ameliorative impacts on the growth and welfare status of Cd-intoxicated Nile tilapia (Oreochromis niloticus). Bifactorial design (3 Bs-CNPs levels × 3 Cd levels) was used in the current study where Nile tilapia fingerlings (58–63 g) were fed on 0.0, 2, and 4 g Bs-CNPs/kg feed alongside with exposing to 0.0, 0.392, and 0.784 mg Cd/L for 60 days to represent nine treatments as follows: T1: control group (no Cd exposure; no Bs-CNPs supplement); T2 and T3: fish were intoxicated with 0.784 and 0.392 mg Cd/L, respectively; T4 and T5: fish fed on 2 and 4 g Bs-CNPs/kg feed, respectively; T6 and T7: fish were fed on 2 g Bs-CNPs/kg feed along with exposure to 0.784 and 0.392 mg Cd/L, respectively; and T8 and T9: fish were fed on 4 g Bs-CNPs/kg feed along with exposure to 0.784 and 0.392 mg Cd/L, respectively. It is noted that the Cd-intoxicated fish exhibited significant retardation in growth performance and digestive enzyme activities with a decline in their survival rate compared to the control group (T1). The results also revealed that exposing fish to Cd toxicity alone with no feed supplement (T2 and T3) experienced abnormal clinical signs and behavioral changes. Compared with the control group (no Cd with no Bs-CNPs), highest values of cortisol, glucose, aspartate and alanine aminotransferase, and acetylcholine esterase activity were found in fish fed on the control diet along with exposure to 0.784 mg Cd/L. Higher Cd restudies in liver, intestine, gills, kidney, and muscles tissues were detected in fish exposed to 0.784 mg Cd/L alone and the sequence order of Cd levels in different fish organs is intestine > gills > liver > kidney > muscles. Remarkable pathological alterations in hepatic and intestinal tissues were also observed. On the other hand, feeding Nile tilapia on Bs-CNPs-enriched diets alone with no Cd exposure enhanced their growth performance, digestive enzyme activities, and hematological parameters with no Cd residues in fish organs. Interestingly, feeding the Cd-intoxicated fish on diets with Bs-CNPs (4 g/kg feed) returned their growth, digestive enzymes, hematological, and biochemical parameters to approximate those of the control group. Furthermore, these treatments showed histopathological alteration recovery in the intestine and liver tissues is similar to those in the control group (no Cd with no Bs-CNPs). Fish fed on Bs-CNPs levels with no Cd exposure showed no Cd residues in different fish organs. The Cd levels in different organs of fish exposed to 0.392 mg Cd/L along with feeding on Bs-CNPs (4 g/kg feed) were lower than those in Cd-exposed fish treatments. Consequently, the current study evoked that feeding Nile tilapia fingerlings on Bs-CNPs (4 g/kg feed) could enhance their growth performance and protect the fish from the Cd toxicity that may occur in the aquatic ecosystem.

Paeoniflorin inhibited GSDMD to alleviate ANIT-induced cholestasis via pyroptosis signaling pathway

BackgroundCholestasis (CT) is a group of disorders caused by impaired production, secretion or excretion of bile. This may result in the deposition of bile components in the blood and liver, which in turn causes damage to liver cells and other tissues. If untreated, CT can progress to severe complications, including cirrhosis, liver failure, and potentially life-threatening conditions. ObjectiveThis research was intended to elucidate the function and mechanism of Paeoniflorin (PF) in ameliorating ANIT-induced pyroptosis in CT. MethodsCT models were established in SD rats and HepG2 cells through ANIT treatment. Histological examination was conducted using haematoxylin and eosin (HE) staining to assess the histopathological alterations in the liver. Network pharmacology was employed to identify potential PF targets in CT treatment. To evaluate pyroptosis levels, various methods were used, including serum biochemical analysis, Enzyme-Linked Immunosorbent Assay (ELISA), immunofluorescence (IF), immunohistochemistry (IHC), Western blotting, transmission electron microscopy (TEM), and scanning electron microscopy (SEM). The HuProt™ 20K Chip was utilized to pinpoint potential PF-binding targets. PF's direct mechanisms in CT treatment were explored using molecular docking (MD), molecular dynamics simulations (MDS), Cellular Thermal Shift Assay (CETSA), and Surface Plasmon Resonance (SPR). ResultsPF administration was found to alleviate ANIT-induced liver pathology, enhance liver function markers, and improve cell viability. Network pharmacology and pyroptosis inhibitor studies suggested that PF might mitigate CT via the NLRP3-dependent pyroptosis pathway. This hypothesis was further supported by Western blotting, IF, and IHC analyses, which indicated PF's potential to inhibit NLRP3-dependent pyroptosis in CT. GSDMD was identified as a target through HuProt™ 20K Chip screening. The binding affinity of PF to GSDMD was validated through MD, MDS, CETSA, and SPR techniques. Additionally, the regulatory impact of GSDMD on downstream inflammatory pathways was confirmed by ELISA and IHC. CONCLUSION: PF exhibited a hepatoprotective effect in ANIT-induced CT, primarily by targeting GSDMD, thereby suppressing ANIT-induced pyroptosis and the subsequent release of inflammatory mediators.

Therapeutic efficacy of JEW-M449, an anti-TCTP monoclonal antibody, administered via the nasal route in a BALB/c mouse model of ovalbumin-induced acute asthma

Numerous studies have highlighted the role of translationally controlled tumor protein (TCTP) as a key inflammatory mediator of asthma and allergies. Our previous study revealed that blocking the cytokine-like activity of TCTP using JEW-M449, an anti-TCTP monoclonal antibody (mAb), alleviated allergic inflammation in asthmatic mice. This study aimed to determine whether directly delivering JEW-M449 into the respiratory tract is a more effective way of mitigating airway inflammation in a mouse model of ovalbumin (OVA)-induced allergic airway inflammation than delivering this antibody via the intraperitoneal (IP) route. OVA-sensitized mice were intranasally administered JEW-M449 to enable its direct delivery to the respiratory tract before OVA challenge. We evaluated the changes in the levels of bronchoalveolar lavage fluid (BALF) cells, T helper type 2 (Th2) cytokines, OVA-specific immunoglobulin E (IgE), and histopathological alterations in the lung tissues. Intranasal (IN) administration of JEW-M449 significantly ameliorated the pathological changes associated with OVA-induced lung injury, including reduced inflammatory cell infiltration and mucus hypersecretion. Mice IN administered JEW-M449 also showed decreased OVA-mediated induction of Th2 cytokines in BALF and lung homogenates. Importantly, JEW-M449 delivered via the IN route reached the lung tissue more effectively and exerted superior anti-inflammatory effects in OVA-challenged mice than the IP-delivered JEW-M449. This study is the first to demonstrate the efficacy of directly delivering JEW-M449 anti-TCTP mAb into the respiratory tract to alleviate the asthma phenotype in a mouse model, thereby highlighting a potential delivery strategy for novel inhaled mAb therapeutics for human asthma.

Subchronic Chloroform Exposure Causes Intestinal Damage and Induces Gut Microbiota Disruption and Metabolic Dysregulation in Mice.

Chloroform is a prevalent toxic environmental pollutant in urban settings, posing risks to human health through exposure via various mediums such as air and tap water. The gut microbiota plays a pivotal role in maintaining host health. However, there is a paucity of research elucidating the impact of chloroform exposure on the gut microbiota. In this investigation, 18 SPF Kunming female mice were stratified into three groups (n = 6) and subjected to oral gavage with chloroform doses equivalent to 0, 50, and 150 mg/kg of body weight over 30 days. Our findings demonstrate that subchronic chloroform exposure significantly perturbs hematological parameters in mice and induces histopathological alterations in cecal tissues, consequently engendering marked disparities in the functional composition of cecal microbiota and metabolic equilibrium of cecal contents. Ultimately, our investigation revealed a statistically robust correlation, exhibiting a high degree of significance, between the intestinal microbiome composition and the metabolites that were differentially expressed consequent to chloroform exposure.

Preclinical Safety Assessment of Lepidium sativum L. Seed Extract and its Nanoparticles via Acute and Subacute Oral Administration

Background: Lepidium sativum (LS) seed extract has various pharmacological properties, such as antioxidant, hepatoprotective, and anticancer activities. However, the translation of L. sativum seed extract to the clinical phase is still tedious due to its bioavailability and stability issues. This problem can be solved by encapsulating it in a nanodelivery system to improve its therapeutic potency. Methods: In this study, we have determined and compared the in vivo toxicity of ethanolic extracts of L. sativum seeds (EELS) and solid lipid nanoparticles (SLNs). To conduct toxicity (acute and subacute toxicity) assessments, EELS and SLNs were orally administered to Swiss albino mice. Animal survival, body weight, the weight of vital organs in relation to body weight, haematological profile, biochemistry profile, and histopathological alterations were examined. Results: Animals administered with 2000 mg/kg and 5000 mg/kg in an acute toxicity study exhibited no toxicological symptoms regarding behaviour, gross pathology, and body weight. As per a study on acute toxicity, the LD50 (lethal dose) for SLNs and EELS was over 400 mg/kg and over 5000 mg/kg, respectively. When animals were given SLNs (50 and 100 mg/kg, orally) and EELS (250, 500, and 1000 mg/kg, orally) for 28 days, subacute toxicity study did not exhibit any clinical changes. There were no differences in weight gain, haematological parameters, or biochemical parameters compared to the control groups (p &gt; 0.05). The organs of the treated animals showed no abnormalities in the histological analysis (liver, heart, kidney, and spleen). Conclusion: The result confirms ethanolic extracts of L. sativum seeds and their SLNs to not have harmful effects following acute and subacute administration to mice. For further studies, patents available on Lepidium may be referred for its preclinical and clinical applications.

Postmortem interval estimation of time since death: impact of non-histone binding proteins, immunohistochemical, and histopathological changes in vivo.

The postmortem interval (PMI) is one of the primary objectives and challenging tasks proposed for determining the time of death. This study aimed to estimate the PMI using serum levels of high mobility group box 1 (HMGB1), a biomarker of pyroptotic cell death, along with desmin immunohistochemical and histological analyses of the gastrocnemius muscle in rats at various time intervals. Serum and gastrocnemius muscle samples were collected at zero, 24-, 48-, 72-, and 96 hours postmortem from 50 rats maintained at 22 ± 2°C. The results revealed that the HMGB1 level peaked at 48 hours and dropped in a time-dependent manner afterward. Immunohistochemical analysis revealed a progressive decrease in desmin expression, with severe immunoreactivity (38.19%) at 0 hours, dropping to a minimal level (1.09%) 96 hours after death. Histological analysis of the gastrocnemius muscle at 96 hours revealed significant vacuolation, loss of normal architecture, reduced nuclear visibility, and complete autolysis of all myocytes. In conclusion, HMGB1 levels, desmin immunoreactivity, and histopathological alterations seen in the gastrocnemius muscle could be helpful, valuable, and potential markers for accurately determining PMIs in humans in future studies.

Histopathological Alterations in the Larvae of Culex pipiens L. (Diptera: Culicidae) Induced by Nanoemulsion of Pimpinella anisum L. Essential Oil

Histopathological Alterations in the Larvae of Culex pipiens L. (Diptera: Culicidae) Induced by Nanoemulsion of Pimpinella anisum L. Essential Oil

SIRT1/NOX1 pathway mediated ameliorative effects of rosmarinic acid in folic acid-induced renal injury.

Renal injury is a serious disorder that can be caused by some diseases or agents. Rosmarinic acid (RA) is a natural and safe compound with powerful antioxidant and anti-inflammatory properties. In this study, the ameliorative effects of RA were assayed in folic acid (FA)-induced renal injury by involving the SIRT1/NOX1 pathway. Thirty-six male C57/BL6 mice were divided into 6 groups (n = 6) including control, vehicle, FA, RA, FA + RA 50, and FA + RA 100. After 10 days, blood urea nitrogen (BUN), creatinine, and oxidative stress were measured. The expression of SIRT1 and NOX1 proteins was evaluated by western blot. Also, histopathological alterations were assayed by H&E and PAS staining methods. BUN and creatinine were significantly higher in the FA group compared to the control group; however, their levels decreased after RA treatment in both doses. A significant decrease was observed in swelling, necrosis, and desquamation of tubular epithelial cells in the FA + RA 50 and FA + RA 100 groups compared to the FA group. RA in the animals receiving FA increased SIRT1 expression and the levels of GSH and SOD compared to the FA group. RA in the animals receiving FA showed a significant decrease in NOX1 expression and MDA level compared to the FA group. The findings declared that the administration of RA has positive effects against renal damage induced by FA. The effect might result from involvement in the SIRT1/NOX1 pathway and thereby attenuation of oxidative stress.

Protective efficacy of thymoquinone against malathion-induced histopathological and biochemical alterations in the gills of Labeo rohita

Protective efficacy of thymoquinone against malathion-induced histopathological and biochemical alterations in the gills of Labeo rohita

Investigation of the pharmacological mechanisms of Shenfu injection in acute pancreatitis through network pharmacology and experimental validation

BackgroundShenfu Injection (SFI) has emerged as a prevalent therapeutic intervention in clinical practice for the management of acute pancreatitis (AP). The purpose of this research was to investigate and validate the potential mechanisms of SFI in the treatment of AP through network pharmacology. MethodsNetwork pharmacology was adopted to investigate the potential targets and mechanisms of SFI in the treatment of AP. Molecular docking was employed to evaluate the binding affinity between active components and targets. Single-cell transcriptome analysis was conducted to explore the cell types associated with SFI treatment in AP. In vitro and in vivo models of AP were induced by caerulein. The histopathological changes were observed by HE staining. Cell apoptosis was detected using flow cytometry and Tunel staining. Cell viability was assessed using CCK-8 assay. Western blot and ELISA were used to detect the protein expression and inflammatory cytokines, respectively. ResultsA total of 104 SFI active components were obtained, of which 29 targeted 76 genes. After intersecting with 3370 AP-related genes, 42 SFI treatment AP potential targets were identified. Enrichment analysis revealed that these targets were associated with cell apoptosis, necroptosis, and multiple signal transduction pathways, such as p53, IL-17 and TNF signal pathways, etc. Molecular docking demonstrated that the active components of SFI had good binding affinity with the corresponding targets and the binding ability of NGF and aromadendrene was the strongest. Bioinformatics analysis revealed that SFI treatment in AP is associated with various cell types, including acinar cells, endothelial cells, T cells, dendritic cells, ductal cells, and mesenchymal cells. Furthermore, in vitro experiments demonstrated that SFI induces acinar cell apoptosis in a dose-dependent manner, accompanied by increased expression of cleaved-caspase3/caspase3 and cleaved-caspase8/caspase8 proteins, and inhibition of inflammatory cytokine (TNF-ɑ, IL-1β, and PTGS2) expression. In vivo experiments demonstrated that SFI improved histopathological alterations, reduces inflammation, and promotes apoptosis and the expression of cleaved-casp3 and cleaved-casp8 in AP rats. ConclusionsThis study elucidated the multi-component, multi-target, and multi-cellular characteristics of SFI in the treatment of AP, and confirmed its mechanism of promoting acinar cell apoptosis.

Active mussel biomonitoring for the health status assessment of the Western Mediterranean Sea

The Western Mediterranean coast is under the influence of anthropogenic pressures, including land use, increasing amounts of dangerous waste and habitat destruction. In 2021, the French RINBIO network (http://www.ifremer.fr/envlit/) originally dedicated to assess chemical contamination in the region, focused on biological effects produced by contaminants and the interaction with natural variability in mussels using an active caging strategy. Cell and tissue level biomarkers were applied for 17 sampling sites divided in three sub-regions categorized by different environmental conditions. Results provide critical information for ecosystem health assessment using mussels as sentinel species in the Western Mediterranean Sea. The influence of natural and confounding factors (trophic condition, reproductive cycle, caging strategy), on biological responses to mild chemical contamination, was discussed and discriminated for health status assessment. Results provide valuable data available as reference values for the assessment of biomarkers and histopathological alterations for large-scale active biomonitoring campaigns in the Western Mediterranean Sea.