Histopathological Scores Research Articles

Parthenolide repressed endometriosis induced surgically in rats: Role of PTEN/PI3Kinase/AKT/GSK-3β/β-catenin signaling in inhibition of epithelial mesenchymal transition

AimPI3K/AKT/GSK-3β/β-catenin signaling pathway is a triggering factor for epithelial to mesenchymal transition (EMT) which plays a pivotal role in the pathogenesis of endometriosis. Parthenolide is a sesquiterpene lactone extract that has anti-inflammatory, analgesic and anticancer properties. Hence, we investigated the effect of parthenolide against EMT in the endometrial tissue implants and immortalized epithelial endometriotic cell lines 12Z. Main methodsTwenty- four female Rats with surgically induced endometriosis were treated with parthenolide (2, 4 mg/kg), for 4 weeks. Endometriotic cell line 12Z was used to identify the effect of parthenolide on the wound healing, cellular migration and invasion properties of endometriotic cells. Key findingsParthenolide decreased the endometriotic implant tissue expression of total PI3K, PI3K-p85, p-AKT, p/total AKT, p-GSK-3β, P/total GSK-3β, and nβ-catenin, as well as increased E-cadherin and decreased vimentin mRNA expression. Parthenolide upregulated PTEN immunoreactivity as well as the endometriotic tissue caspase-3, caspase-9, BAX levels while reducing Bcl2 level. Additionally, parthenolide decreased endometriotic tissue implants surface area and histopathological score of the epithelial growth. SignificanceOur findings showed that parthenolide in a dose dependent manner inhibited PI3K/AKT/GSK-3β/nβ-catenin cascade via enhancement of PTEN with subsequent inhibition of EMT evidenced by elevation of the epithelial marker, E-cadherin and reduction of mesenchymal marker, vimentin, of the endometriotic implants in addition to reversal of invasion and migration properties of epithelial endometriotic cell lines. These findings provide a valuable therapeutic approach for treatment of endometriosis.

Correlation analysis of quantitative MRI measurements of thigh muscles with histopathology in patients with idiopathic inflammatory myopathy

ObjectivesTo validate the correlation between histopathological findings and quantitative magnetic resonance imaging (qMRI) fat fraction (FF) and water T2 mapping in patients with idiopathic inflammatory myopathy (IIM).MethodsThe study included 13 patients with histopathologically confirmed IIM who underwent dedicated thigh qMRI scanning within 1 month before open muscle biopsy. For the biopsied muscles, FF derived from the iterative decomposition of water and fat with echo asymmetry and least-squares estimation quantitation (IDEAL-IQ) and T2 time from T2 mapping with chemical shift selective fat saturation were measured using a machine learning software. Individual histochemical and immunohistochemical slides were evaluated using a 5-point Likert score. Inter-reader agreement and the correlation between qMRI markers and histopathological scores were analyzed.ResultsReaders showed good to perfect agreement in qMRI measurements and most histopathological scores. FF of the biopsied muscles was positively correlated with the amount of fat in histopathological slides (p = 0.031). Prolonged T2 time was associated with the degree of variation in myofiber size, inflammatory cell infiltration, and amount of connective tissues (p ≤ 0.008 for all).ConclusionsUsing the machine learning-based muscle segmentation method, a positive correlation was confirmed between qMRI biomarkers and histopathological findings of patients with IIM. This finding provides a basis for using qMRI as a non-invasive tool in the diagnostic workflow of IIM.Relevance statementBy using ML-based muscle segmentation, a correlation between qMRI biomarkers and histopathology was found in patients with IIM: qMRI is a potential non-invasive tool in this clinical setting.Key points• Quantitative magnetic resonance imaging measurements using machine learning-based muscle segmentation have good consistency and reproductivity.• Fat fraction of idiopathic inflammatory myopathy (IIM) correlated with the amount of fat at histopathology.• Prolonged T2 time was associated with muscle inflammation in IIM.Graphical

Luteolin inhibits herpes simplex virus 1 infection by activating cyclic guanosine monophosphate-adenosine monophosphate synthase-mediated antiviral innate immunity

BackgroundThe successive outbreaks of large-scale infectious diseases due to virus infection have been a major threat to human health in recent decades. Herpes simplex virus I (HSV-1) is a widely-disseminated DNA virus that infects the central nervous system to cause herpes labialis, keratitis and herpes simplex virus encephalitis (HSE), resulting in recurrent lifelong clinical or subclinical episodes. Luteolin is a plant flavone that has been extensively used in the treatment of various human diseases, including carcinogenesis, inflammation and chronic degenerative diseases. PurposeThe aim of this study was to investigate the antiviral molecular mechanism of luteolin against HSV-1 infection in vitro and in vivo. MethodsThe antiviral effect of luteolin in cell lines was examined by viral plaque assay, RT-qPCR, Western blot and time-of-addition assay. The interaction between luteolin and cyclic guanosine monophosphate-adenosine monophosphate synthase (cGAS) was evaluated by molecular modeling and semi-denaturing detergent agarose gel electrophoresis. The efficacy of luteolin on HSE was evaluated in the HSE mouse model by analyzing weight loss, neurodegenerative symptoms and histopathological scores. Cytokine expression and protein levels were examined by RT-qPCR, Western blot and ELISA. ResultsLuteolin inhibited the early process of HSV-1 infection, without affecting the infection of acyclovir-resistant HSV-1 strains. In addition, luteolin enhanced antiviral type I interferon production and activated the cytoplasmic DNA-sensing cGAS-stimulator of interferon gene (STING) pathway. Luteolin directly bound the active substrate binding site and promoted the oligomerization of cGAS. Luteolin also inhibited HSE-related weight loss, neurodegeneration and neuroinflammation in mice caused by HSV-1 infection. Furthermore, luteolin enhanced type I interferon expression and stimulated the cGAS-STING signaling pathway in vivo. ConclusionLuteolin inhibited the post-entry process of HSV-1 by activating the cGAS-STING pathway to promote antiviral interferon production. These results provided the rationale for luteolin as a potent cGAS activator and antiviral agent.

Brensocatib (an oral, reversible inhibitor of dipeptidyl peptidase-1) attenuates disease progression in two animal models of rheumatoid arthritis.

Rheumatoid arthritis (RA) is a painful and incurable disease characterized by chronic joint inflammation and a progressive destruction of cartilage and bone. Although current treatments have improved clinical outcomes for some patients, the high relapse rates and sizeable proportion of non-responders emphasize the need for further research. Arthritic joints are massively infiltrated by neutrophils, which influence inflammatory and immune processes by releasing cytokines, chemokines, eicosanoids, and neutrophil serine proteases (NSPs) - all of which are known to contribute to RA initiation and progression. Active NSPs are generated from zymogens at the promyelocytic stage of neutrophil differentiation under the action of dipeptidyl peptidase 1 (DPP-1) and DPP-1 knockout mice are resistant to the development of arthritis. Thus, DPP-1 inhibition represents a promising therapeutic approach in RA. In this study, we assessed the efficacy of a potent and highly selective DPP-1 inhibitor, brensocatib, in two well established RA models - rat collagen-induced arthritis (CIA) and mouse collagen antibody-induced arthritis (CAIA). In both models, brensocatib at 3 and 30 mg/kg/day significantly reduced bone marrow NSP levels, in keeping with prior pharmacodynamic studies in rodents. More importantly, brensocatib treatment significantly improved disease score at both dosages in both rodent models. In the mouse CAIA model, brensocatib even proved at least as potent as anti-TNF antibodies in diminishing both the histopathological score and neutrophil infiltration into arthritic joints. Together, these results show that brensocatib alters RA disease progression in rodents and supports the need for its further evaluation as a potential therapeutic option, or to complement existing RA treatments.

Prevalence and In Vivo Assessment of Virulence in Shiga Toxin-Producing Escherichia coli Clinical Isolates from Greater Cairo Area.

Background: Shiga toxin-producing Escherichia coli (STEC) has been identified as an important etiologic agent of human disease in Egypt. Aims: To investigate the occurrence and describe the characterization as well as prevalence of STEC in Greater Cairo hospitals as well as molecular characterization of virulence and resistance genes. Methods: Four hundred seventy E. coli clinical isolates were collected from eight hospitals and analyzed by genotypic and phenotypic methods for STEC, followed by histopathological examination and scoring of different organs lesions. Results: The highest proportion of isolates was from urine (151 isolates), whereas the lowest was from splenic drain (3 isolates). In tandem, when serogrouping was performed, 15 serogroups were obtained where the most prevalent was O157 and the least prevalent was O151. All isolates were positive when screened for identity gene gad A, while only typable strains were screened for seven virulence genes stx1 (gene encoding Shiga toxin 1), stx2 (gene encoding Shiga toxin 2), tsh (gene encoding thermostable hemagglutinin), eaeA (gene encoding intimin), invE (gene encoding invasion protein), aggR (gene encoding aggregative adherence transcriptional regulator), and astA (aspartate transaminase) where the prevalence was 48%, 30%, 50%, 57%, 7.5%, 12%, and 58%, respectively. Of 254 typable isolates, 152 were STEC carrying stx1 or stx2 genes or both. Conclusions: Relying on in vivo comparison between different E. coli pathotypes via histopathological examination of different organs, E. coli pathotypes could be divided into mild virulent, moderate virulent, and high virulent strains. Statistical analysis revealed significant correlation between different serogroups and presence of virulence genes.

A new rat model of chronic relapsing ulcerative colitis with traditional Chinese medicine syndrome of yang deficiency and dampness stagnation

IntroductionChronic relapsing ulcerative colitis (CRUC) is classified as an inflammatory bowel disease, and traditional Chinese medicine (TCM) has been recognized for its therapeutic benefits in treating CRUC. According to TCM principles, the prevailing syndromes in CRUC are yang deficiency and dampness stagnation (YDDS). The objective of this study is to establish a novel animal model of CRUC exhibiting the YDDS syndrome. MethodsInitially, the rats were administered hydrocortisone succinate sodium through subcutaneous injection, followed by the administration of senna folium granule solution. Subsequently, the rats underwent three cycles of dextran sulfate sodium solution ingestion. ResultsUpon completion of the modeling process, it was observed that the disease activity index, colonic mucosal damage index score, colon weight/length ratio, colonic histopathology score, and tissue damage index scores of the model rats were elevated, while the colon length and spleen index exhibited a decrease compared to the control group. The pathological alterations in the model rats included the presence of shallow or deep ulcers and acute or chronic inflammation throughout the process of CRUC modeling. Additionally, our findings indicate a correlation between the proximity of the ulcer to the anal verge, the severity of the lesion, and the presence of precancerous glands, glandular disappearance, crypt abscesses, and tissue destruction. Furthermore, the expression levels of IL6, IL10, IFNG, and TNFA in the colon tissues were significantly elevated in the model rats compared to the control group. Notably, the end-stage lesions observed in CRUC exhibited greater severity when compared to those observed in cases of acute ulcerative colitis. DiscussionIn conclusion, we have successfully developed a rat model of CRUC with YDDS syndrome, which can be utilized to investigate the effectiveness and underlying mechanisms of TCM in treating CRUC.

Establishment of a Mycoplasma hyorhinis challenge model in 5-week-old piglets.

Mycoplasma hyorhinis is an emerging swine pathogen with high prevalence worldwide. The main lesions caused are arthritis and polyserositis, and the clinical manifestation of the disease may result in significant economic losses due to decreased weight gain and enhanced medical costs. We aimed to compare two challenge routes to induce M. hyorhinis infection using the same clinical isolate. Five-week-old, Choice hybrid pigs were inoculated on 2 consecutive days by intravenous route (Group IV-IV) or by intravenous and intraperitoneal routes (Group IV-IP). Mock-infected animals were used as control (control group). After the challenge, the clinical signs were recorded for 28 days, after which the animals were euthanized. Gross pathological and histopathological examinations, PCR detection, isolation, and genotyping of the re-isolated Mycoplasma sp. and culture of bacteria other than Mycoplasma sp. were carried out. The ELISA test was used to detect anti-M. hyorhinis immunoglobulins in the sera of all animals. Pericarditis and polyarthritis were observed in both challenge groups; however, the serositis was more severe in Group IV-IV. Statistically significant differences were detected between the challenged groups and the control group regarding the average daily weight gain, pathological scores, and ELISA titers. Additionally, histopathological scores in Group IV-IV differed significantly from the scores in the control group. All re-isolated strains were the same or a close genetic variant of the original challenge strain. Our results indicate that both challenge routes are suitable for modeling the disease. However, due to the evoked more severe pathological lesions and the application being similar to the hypothesized natural route of infection in Group IV-IV, the two-dose intravenous challenge is recommended by the authors to induce serositis and arthritis associated with M. hyorhinis infection.

Establish and apply ISI pathological method to evaluate the intestinal health of largemouth bass (Micropterus salmoides) and channel catfish (Ictalurus punctatus) challenged with Aeromonas hydrophila

Gut health is essential for animal growth because it can absorb nutrients more efficiently, promote growth, and protect the body from invasion by pathogenic microorganisms. Temperature, dissolved oxygen, water quality, nutrition level, pathogen, net fishing, and so on will affect the gut health of aquatic animals. Intestinal diseases in aquatic animals were often not easily detected, making them difficult to prevent and control. Therefore, establishing a stable and scientific evaluation standard of gut health was of great significance. This study evaluated the effects of gross pathological and histological parameters of largemouth bass and channel catfish infected by Aeromonas hydrophila. A total of 150 largemouth bass and 150 channel catfish were randomly divided into 5 groups: 1 control group (NC) and 4 challenge groups (CH, fish were infected with A. hydrophila by intraperitoneal injection with a concentration of 200 µL 1x108 CFU/mL, 1x107 CFU/mL, 1x106 CFU/mL, and 1x105 CFU/mL, respectively). After the challenge, largemouth bass’ gross pathological and histological changes were observed on the 2nd, 6th, 9th, 12th, and 14th days. In contrast, channel catfish’s gross pathological and histological changes were observed on the 3rd, 6th, 9th, 12th, and 15th day. In this study, regarding the “I See Inside” (ISI) methodology of poultry, a pathological evaluation method of fish intestinal health was established to comprehensively evaluate the intestinal health status of fish by gross pathological and histopathological score. In gross pathology, it was scored by the body surface and intestinal lesions. In histology, it was a metric evaluation of histological alterations in the intestine, converting macroscopic and microscopic alterations into numbers and then scoring. Each evaluation parameter sets up an “influence factor” (IF) according to the degree of influence of its pathological changes on intestinal function. The final score was the sum of the IF product and this parameter’s score. The results of two kinds of fish showed that the gross pathological and histopathological scores correlated with the infection dose and time. And the significant difference in the score between the enteritis and control groups was observed. The results showed that the higher score, the more serious the pathological changes of fish infected by A. hydrophlia. From this, it can be concluded that the pathological evaluation method established can be used to evaluate the gut health of fish.

The potential renoprotective effect of Raloxifene in renal ischemia-reperfusion injury in a male rat model.

Renal ischemia-reperfusion injury is caused by a temporary reduction in oxygen-carrying blood flow to the kidney, followed by reperfusion. During ischemia, kidney tissue damage induces overproduction of reactive oxygen species, which produces oxidative stress. The blood flow restoration during the reperfusion period causes further production of reactive oxygen species that ends with apoptosis and cell death. This study aimed to investigate the potential renoprotective effects of Raloxifene on bilateral renal ischemia-reperfusion injury in rats by looking into kidney function biomarkers, urea and creatinine, inflammatory cytokines, such as tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β). Additionally, antioxidant markers such as total antioxidant capacity (TAC) and the pro-apoptotic marker caspase-3 were assessed. Histopathological scores were also employed for evaluation. Our experimental design involved 20 rats divided into four groups: the sham group underwent median laparotomy without ischemia induction, the control group experienced bilateral renal ischemia for 30 minutes followed by 2 hours of reperfusion, the vehicle group received pretreatment with a mixture of corn oil and dimethyl sulfoxide (DMSO) before ischemia induction, and the Raloxifene-treated group was administered Raloxifene at a dose of 10 mg/kg before ischemia induction, followed by ischemia-reperfusion. Urea and creatinine, TNF-α, IL-1β, and caspase-3 in the Raloxifene group were significantly lower compared to the control and vehicle groups. On the other hand, TAC levels in the Raloxifene group were significantly higher than in the control and vehicle groups. This study concluded that Raloxifene had a renoprotective impact via multiple actions as an anti-inflammatory, anti-apoptotic, and antioxidant agent.

Native T1 mapping for non-invasive quantitative evaluation of renal function and renal fibrosis in patients with chronic kidney disease.

To investigate the role of native T1 mapping in the non-invasive quantitative assessment of renal function and renal fibrosis (RF) in chronic kidney disease (CKD) patients. A prospective analysis of 71 consecutive patients [no RF (0%): 9 cases; mild RF (<25%): 36 cases; moderate RF (25-50%): 17 cases; severe RF (>50%): 9 cases] who were clinically diagnosed with CKD that was pathologically confirmed and who underwent magnetic resonance imaging (MRI) examination between October 2021 and September 2022 was performed. T1-C (mean cortical T1 value), T1-M (mean medullary T1 value), ΔT1 (mean corticomedullary difference) and T1% (mean corticomedullary ratio) values were compared. Correlations between T1 parameters and clinical and histopathological values were analyzed. Regression analysis was performed to determine independent predictors of RF. The areas under the receiver operating characteristic curve (AUC) were calculated to assess the diagnostic value of RF. The T1-C, ΔT1 and T1% values (P<0.05) were significantly different in the CKD group, but T1-M was not (P>0.05). The ΔT1 and T1% values showed significant differences in pairwise comparisons among CKD subgroups (P<0.05) except for CKD 2 and 3. ΔT1 and T1% were moderately correlated with the estimated glomerular filtration rate (ΔT1: rs=-0.561; T1%: r=-0.602), serum creatinine (ΔT1: rs=0.591; T1%: rs=0.563), blood urea nitrogen (ΔT1: rs=0.433; T1%: rs=0.435) and histopathological score (ΔT1: rs=0.630; T1%: rs=0.658). ΔT1 and T1%, but not T1-C, were independent predictors of RF (P<0.05). ΔT1 and T1% were set as -410.07 ms and 0.8222 with great specificity [ΔT1: 91.7% (77.5-98.2%); T1%: 97.2% (85.5-99.9%)] to identify mild RF and moderate-severe RF. The optimal cutoff values for differentiating severe RF from mild-moderate RF were -343.81 ms (ΔT1) and 0.8359 (T1%) with high sensitivity [both 100% (66.4-100%)] and specificity [ΔT1: 90.6% (79.3-96.9%); T1%: 94.3% (84.3-98.8%)]. ΔT1 and T1% overwhelm T1-C for assessment of renal function and RF in CKD patients. ΔT1 and T1% identify patients with <25% and >50% fibrosis, which can guide clinical decision-making and help to avoid biopsy-related bleeding.

Systemic administration of budesonide in pegylated liposomes for improved efficacy in chronic rhinosinusitis.

Chronic rhinosinusitis (CRS) is a chronic inflammatory condition affecting the nasal and paranasal sinuses of approximately 11.5% of the United States adult population. Oral corticosteroids are effective in controlling sinonasal inflammation in CRS, but the associated adverse effects limit their clinical use. Topical budesonide has demonstrated clinical efficacy in patients with CRS. Herein, we investigated the systemic delivery of liposomes tethered with poly(ethylene glycol) (PEG) and loaded with budesonide in a murine model of CRS. PEGylated liposomes encapsulated with budesonide phosphate (L-BudP) were administered via tail vein injection, and the feasibility of L-BudP to reduce sinonasal inflammation was compared to that of free budesonide phosphate (F-BudP) and topical budesonide phosphate (T-BudP) treatment over a 14-day study period. Compared to a single injection of F-BudP and repeat T-BudP administration, a single injection of L-BudP demonstrated increased and prolonged efficacy, resulting in the significant improvement of sinonasal tissue histopathological scores (p<0.05) with decreased immune cell infiltration (p<0.05). Toxicities associated with L-BudP and T-BudP treatment, assessed via body and organ weight, as well as peripheral blood liver enzyme and differential white blood cell analyses, were transient and comparable. These data suggest that systemic liposomal budesonide treatment results in improved efficacy over topical treatment.

Early Life Exposure to Human Milk Oligosaccharides Reduces Allergic Response in a Murine Asthma Model.

Studies suggest that early-life gut microbiota composition and intestinal short-chain fatty acids (SCFAs) are linked to future asthma susceptibility. Furthermore, infancy offers a critical time window to modulate the microbiota and associated metabolites through diet-microbe interactions to promote infant health. Human milk oligosaccharides (HMOs), nondigestible carbohydrates abundant in breast milk, are prebiotics selectively metabolized by gut microbiota that consequently modify microbiome composition and SCFA production. Using a house dust mite mouse model of allergy, we investigated the impacts of early oral treatment of pups with biologically relevant doses of 2'-fucosyllactose (2'-FL) and 6'-sialyllactose (6'-SL), two of the most abundant HMOs in human milk, in amelioration of allergic airway disease severity. We found that administration of 2'-FL and 6'-SL during early life reduced lung histopathology scores, circulating IgE, cytokine levels, and inflammatory cell infiltration, all hallmark symptoms of allergic asthma. HMO supplementation also increased the relative abundance of intestinal Bacteroidetes and Clostridia, known SCFA producers within the gut. Indeed, we detected increased SCFA concentrations in both the intestine and blood of adult mice who received HMOs prior to weaning. We propose a model in which orally administered HMOs delivered during early life shift the microbiota toward increased production of SCFAs, which dampens the allergic immune responses behind allergy and asthma. Overall, these data suggest the potential for HMO supplementation to protect infants against asthma development later in life, with possible benefits against additional atopic diseases such as eczema and food allergies.

Role of histopathology in severity assessments of post-COVID-19 rhino-orbital cerebral mucormycosis - A case-control study

BackgroundThere was an upsurge in rhino-orbital-cerebral mucormycosis (ROCM) during the second wave of the COVID-19 pandemic in India. Histopathological examination of the biopsies witnessed greater tissue invasion in these cases. The present study aimed to evaluate the histopathological scoring system in the severity assessment and to compare the scores between ROCM that required orbital exenteration (cases) and those that were treated conservatively (controls). Material and methodsA retrospective study was conducted to analyze the histopathological features using a scoring system in patients diagnosed with mucormycosis on sino nasal biopsies. The scoring was based on the following parameters: the extent of necrosis (score 1 <50% and score 2 >50%), degree of angioinvasion (score 1 <3 vessels in 10 fields and score 2 > 3 vessels in 10 fields), degree of inflammation (mild, moderate or severe), and fungal load(score 1 to 4), based on which three grades (I to III) were given. The scores were compared between the case group and the control group. The data obtained were analyzed statistically. ResultsThe study included 40 controls and 10 cases. The median age of patients in the control group was 52 with 82.5%males (n = 43) and 17.5 % females (n = 7) while in the case group, the median age was 51 years, and all were males. The predominant inflammatory response was of mixed suppurative type (70 %, n = 28). The degree of necrosis was scored 2 in 57.5 % (n = 23) of controls and 70 %(n = 7) in the case group. All the samples in the case group showed scores of 2 for angioinvasion and scores of 3 and 4 for fungal load. There was a statistically significant difference in the degree of necrosis, angioinvasion, the severity of neutrophilic infiltrates, and fungal load between the cases and controls (p-value <0.05). ConclusionThe present study analyses the predictive role of histopathology in invasive mucormycosis. Higher fungal load, degree of angioinvasion, and the absence of granulomas are associated with advanced disease in sinonasal mucormycosis. The study findings recommend incorporating the histopathological scoring criteria in evaluating patients with sinonasal mucormycosis.

Live IBD vaccine exacerbates disease and pathological effects of Asian lineage H9N2 LPAIV in chickens

ABSTRACT Avian influenza H9N2 is one of the most commonly circulating viruses in numerous Egyptian poultry farms. The Asian lineage H9N2 exhibits an immunosuppressive effect, and its pathogenicity is amplified when it co-infects with other pathogens, especially with the immunosuppressive infectious bursal disease virus (IBDV), resulting in increased mortality rates. Both vaccines and field infection can exacerbate the pathogenicity of H9N2, particularly in the bursa of Fabricius, causing more significant lymphoid depletion. To comprehend the impact of the IBD vaccine on the viral and pathogenic effect of H9N2 infection in specific pathogen-free chicks (SPF), the experiment was designed as four groups; group 1 served as the negative control, group 2 received (228E) IBD vaccine, group 3 was challenged with H9N2, and group-4 was vaccinated by the IBD vaccine then challenged with H9N2. The clinical signs, relative immune organs weights and histopathological lesion scores were recorded. The tracheal and cloacal H9N2 viral shedding were also measured. Group 4 exhibited a significant decrease (P ≤ 0.05) in the relative bursal weight and an increase in the bursal lesion score when compared with groups 1 and 3 at 4 and 8 days post-challenge (dpc). The tracheal lesion score of group-4 recorded a significant increase when compared with groups 1 and 3. The renal lesion score of group 4 achieved a significant increase when compared with 1 and 3 at 8 dpc. Also, group 4 recorded a significant increase in H9N2 shedding in comparison with groups 1 and 3. Consequently, our study concluded that routine vaccination with the IBD intermediate plus vaccine exacerbates the silent infection of H9N2 resulting in outbreaks.

The curative effect of ozonated bidistilled water on Escherichia coli-induced endometritis in rats.

Endometritis is a prevalent and challenging condition that affects livestock, and its effective treatment is the use of intrauterine antibiotics. Antibiotic use may lead to negative consequences, including residue in tissues and the emergence of antibiotic resistance. The aim of this study was to assess the therapeutic potential of ozonated bidistilled water in the treatment of Escherichia coli-induced endometritis in rat models, comparing it to conventional antibiotic treatment. A total of 21 female Wistar Albino rats were used for this study. E. coli from the uterus of cows with endometritis at a concentration of 1.0×109 cfu/mL inoculated into rat uteruses. After inoculation, it was confirmed macroscopically that experimental endometritis was induced in all individuals. And, the rats with endometritis were randomly divided into three groups treated with intrauterine bidistilled water enriched with ozone, antibiotics and 0.9% NaCl. The uterine tissues were examined histopathologically. In the semiquantitative analyses, significantly lower histopathological scores were observed for both the bidistilled water enriched with ozone and antibiotic-treated endometritis groups compared to the control group (p < 0.05). Ozone-enriched bidistilled water can be used in the treatment of experimentally induced endometritis in rats, and can provide effective microbiological and histopathological improvement. Further studies involving larger populations of different species are needed to determine the medical suitability of this new treatment.

Evaluation and comparison of pharmacokinetic profiles and safety of two extended-release buprenorphine formulations in common marmosets (Callithrix jacchus)

While sustained-release buprenorphine (BSR) is used as a long-lasting opioid analgesic in common marmosets (Callithrix jacchus), there are no published studies on pharmaceutical-grade extended-release buprenorphine options such as Ethiqa XR (EXR) for this species. However, BSR is a compounded product and has been reported to cause injection site reactions in multiple species, including marmosets. Additionally, now with the availability of EXR, a pharmaceutical-grade veterinary product, the use of BSR in laboratory animals is not compliant with the Guide for the Care and Use of Laboratory Animals (Guide) unless scientifically justified and approved by the IACUC. We compared pharmacokinetic and safety profiles of BSR (0.15 mg/kg) and EXR (0.1–0.2 mg/kg) administered subcutaneously to adult marmosets. Blood was collected by venipuncture of the saphenous vein at multiple time points (0.25–72 h) and analyzed by liquid chromatography-tandem mass spectrometry (LC–MS/MS). EXR between 0.1 and 0.2 mg/kg resulted in a dose-dependent increase in Cmax (1.43–2.51 ng/mL) and were not statistically different from BSR (1.82 ng/mL). Tmax, lambdaz, and t1/2 were not statistically different between formulations. Mean plasma buprenorphine concentrations for BSR and EXR exceeded the therapeutic threshold (0.1 ng/mL) within 0.25 h and lasted for > 72 h. Mild sedation, but neither respiratory depression nor ataxia, was observed for both formulations. BSR injection sites had significantly higher histopathological scores compared to EXR. Video recordings for monitoring drug-induced behavioral changes showed increased animal activity levels after BSR and EXR versus saline controls. Norbuprenorphine, a buprenorphine metabolite associated with respiratory depression, was detected in the plasma after BSR and EXR administration as well as by in vitro liver microsome assays. In conclusion, we recommend using EXR over BSR as a long-lasting buprenorphine analgesic in marmosets because EXR is a pharmaceutical-grade formulation that is compliant with FDA guidelines and the Guide as well as exhibits comparable PK and safety profiles as BSR.

Therapeutic Effects of Valeriana jatamansi on Ulcerative Colitis: Insights into Mechanisms of Action through Metabolomics and Microbiome Analysis.

Ulcerative colitis (UC), belonging to inflammatory bowel disease (IBD), is a chronic and relapsing inflammatory disorder of the gastrointestinal tract, which has not been completely cured in patients so far. Valeriana jatamansi is a Chinese medicine used clinically to treat "diarrhea," which is closely related to UC. This study was to elucidate the therapeutic effects of V. jatamansi extract (VJE) on dextran sodium sulfate (DSS)-induced UC in mice and its underlying mechanism. In this work, VJE effectively ameliorates the symptoms and histopathological scores and reduces the production of inflammatory factors in UC mice. The colon untargeted metabolomics analysis and 16S rDNA sequencing showed remarkable differences in colon metabolite profiles and intestinal microbiome composition between the control and DSS groups, and VJE intervention can reduce these differences. Thirty-two biomarkers were found and modulated the primary pathways including pyrimidine metabolism, arginine biosynthesis, and glutathione metabolism. Meanwhile, twelve significant taxa of gut microbiota were found. Moreover, there is a close relationship between endogenous metabolites and intestinal flora. These findings suggested that VJE ameliorates UC by inhibiting inflammatory factors, recovering intestinal maladjustment, and regulating the interaction between intestinal microbiota and host metabolites. Therefore, the intervention of V. jatamansi is a potential therapeutic treatment for UC.

Two Age Groups of Adult Pikeperch (Sander lucioperca) as Bioindicators of Aquatic Pollution

Pikeperch of age classes 3+ and 4+ were collected from the Garaši reservoir (Serbia) to analyze their bioindicator potential and compare their possible differences. Concentrations of 26 elements were determined in gills, muscles, and liver by inductively-coupled plasma optical emission spectrometry (ICP-OES) and of 17 organochlorine pesticides (OCPs), and six polychlorinated biphenyls (PCBs) in muscle by gas chromatography with mass spectrometric detection (GC-MS). Histopathological changes in the liver and gills were analyzed as biomarkers of general fish health. Only the concentrations of Cd, Na, and P in the muscles differed significantly. The OCPs and PCBs concentrations were below the detection limits, so fish meat consumption does not pose a risk concerning these substances. Hg and Cd exceeded the maximum allowed concentrations in some 4+ individuals, probably due to biomagnification. Gills were the most affected by metal exposure in both age classes. Histopathological changes and indices were minor and did not differ significantly between age classes, suggesting that pollution did not affect the morphology and structure of gills and liver. There were no significant correlations between elemental accumulation and fish condition or between histopathological scores. Therefore, both age classes can be used as bioindicators of pollution.

Anti-phospholipase A2 receptor positive membranous nephropathy: investigating the link between electron microscopy stages and clinical outcome

ABSTRACT This retrospective, observational study sought to examine the relationship between Ehrenreich-Churg electron microscopy (EM) stages and long-term outcomes in anti-PLA2R membranous nephropathy (MN). Seventy-one patients with anti-PLA2R MN (median titer 185.7RU/mL) were followed for a median of 46 months, with end-stage kidney disease (ESKD) as the primary endpoint, and response to treatment as a secondary endpoint. Patients were grouped into stages I-II (41 patients) and stages III-IV (30 patients) for analytical purposes. Notably, the III-IV group demonstrated a lower eGFR, lower anti-PLA2R titer, but a higher chronicity score. Kaplan-Meier analysis showed shorter mean kidney survival time in stages III-IV compared to I-II (p 0.03). However, multivariate analysis using Cox regression indicated that Ehrenreich-Churg stages did not significantly influence kidney survival, but lower eGFR at diagnosis and higher histopathological chronicity score did. Remission was achieved by 64% of patients and no relationship between Ehrenreich-Churg stages and treatment response was found. The only identified risk factor for not achieving remission was the severity of hyposerinemia at diagnosis. In conclusion, while EM stages III-IV are associated with more chronic lesions and stages I-II with more active immunologic disease, the histological chronicity score seems to be a stronger predictor of long-term outcomes.

Mesenchymal stem cells derived from bone marrow and adipose tissue for repairing acute sciatic nerve injury in a rabbit model

Peripheral nerve injury is one of the common disabling clinical conditions and around 50% of the cases end up in permanent impairment. Due to the lack of effective treatment options regenerative medicine employing stem cells is being evaluated. The presented study evaluated and compared regeneration potential of mesenchymal stem cells (MSCs) derived from bone marrow (BM) and adipose tissue (AD) in acute rabbit sciatic nerve injury (axonotmesis) model. A total of n = 54 grey giant rabbits were made subject of the study and divided equally into 3 groups: Control, BM-MSCs in Collagen I and AD-MSCs in Collagen I as per the treatment given. Iliac crest BM and omental AD was harvested from the same donor for isolation and culture of MSCs. The repair of sciatic nerve injury was evaluated on days 60 and 90. The clinical and histopathological scores and SEM morphology was better in cell treated groups as compared to the control. Morphology and histological studies revealed injured nerve in different levels of regenerative process. Gene expression was more than double for N-Cadherin in cell treated groups as compared to the control, especially at day 60. Between cell treated groups, BM-MSCs group showed better response as compared to the AD-MSCs, although statistically non-significant (p > 0.05). Incomplete nerve regeneration observed under various diagnostic parameters was in compliance to the incomplete clinical recovery at day 90. It was concluded that MSCs may improve sciatic nerve healing but fall short of complete regeneration at day 90, although BM-MSCs may have an edge over AD-MSCs.