Histopathological Approaches Research Articles

Protective effects of some antioxidants against long-term intake of aspartame toxicity on liver and kidney: biochemical and histopathological approach in rats

BackgroundAspartame is used to treat obesity, and the diabetic people could induce changes in liver and kidney structures and function. Garlic extract, melatonin and thymoquinone have an important role against aspartame toxicity due to their antioxidant properties. The current study was designed to examine the protective effects of garlic, melatonin and thymoquinone against aspartame-induced hepatorenal toxicity in albino rates.ResultsThe results indicated that aspartame induced changes in the serum levels of liver parameters function (glucose, ALT, AST and ALP) and renal parameters function (urea and creatinine). Garlic, melatonin and thymoquinone reversed the values of liver and kidney enzymes levels near to or similar to that of control. The histopathological effects of aspartame on the histological structures of liver and kidney were either reduced or removed by garlic, melatonin and thymoquinone.ConclusionLong-term (6 months) administration of aspartame induced toxic effects on hepatorenal function and structure, whereas garlic, melatonin and thymoquinone resulted in hepatorenal ameliorative and protective effects.

Pathology reporting of margin status in locally advanced pancreatic cancer: challenges and uncertainties.

Chemo(radio)therapy is becoming the new standard for patients with locally advanced pancreatic cancer. In case of tumor regression on imaging, surgical resection can be undertaken, albeit often with the need for extended procedures. Reevaluation of the current routine pathology procedures is required to establish the appropriate histopathological approach of the resulting specimens. This review focusses on margin status, which is universally considered a core data item of the pathology report, of relevance to both the management of the individual patient and the evaluation of the result of surgery in this particular patient group. As explained in this review, due to the cytoreductive effect of neoadjuvant therapy, the conventional definition of a tumor-free margin ("R0") based on 1 mm clearance is not adequate. Furthermore, the complexity of many of the specimens following extended or multivisceral en bloc surgical resection make margin assessment challenging. These large specimens require extensive sampling, which is not always easily implemented in daily practice. At present, there is marked divergence in pathology practice, and consequently, neither the true R0-rate nor the exact prognostic effect of the margin status have been definitively established for resected locally advanced pancreatic cancer. A concerted effort towards uniform and optimal margin assessment is unfortunately still lacking.

Interferon-γ-Induced Myeloid-Derived Suppressor Cells Aggravate Kidney Ischemia-Reperfusion Injury by Regulating Innate Immune Cells

Objective: Myeloid-derived suppressor cells (MDSCs) are heterogeneous cells which can suppress T-cell functionality. Herein, we evaluated the functional importance of MDSCs in the context of kidney ischemia-reperfusion injury (IRI) and explored their ability to regulate innate and adaptive immune cell function in this context. Methods: The differentiation of MDSCs was induced in vitro by treating cells with GM-CSF and interferon (IFN)-γ. In a murine model of renal IRI, serum creatinine and blood urea nitrogen values were measured to monitor kidney function, while histopathological and immunohistochemical approaches were used to assess kidney injury severity. In addition, flow cytometry was employed to assess the phenotypes and apoptosis of kidney cells in these mice. Results: MDSCs induced by treatment with GM-CSF + IFN-γ could suppress T-cell functionality in vitro. The adoptive transfer of these MDSCs into an IRI mouse model system enhanced kidney damage and impaired renal function following the recruitment of these cells to renal tissues in these mice. Following such adoptive transfer, the relative frequency of MDSCs with a CD11b⁺Ly6G⁻Ly6C^high monocytic-MDSC phenotype decreased, whereas cells with a CD11b⁺Ly6G⁺Ly6C^low polymorphonuclear-MDSC phenotype become more prevalent within kidney tissues following IRI. Adoptive transfer also coincided with increased frequencies of macrophages and dendritic cells (DCs) in the kidney tissues. This suggested that M-MDSCs contributed to early-stage renal IRI damage by differentiating into these deleterious cell types. However, MDSC-induced suppression of CD4⁺ and CD8⁺ T-cell infiltration was not sufficient to prevent the deterioration of renal function in these mice. Conclusions: Herein, we successfully developed a protocol wherein MDSCs were differentiated in vitro through combination GM-CSF/IFN-γ treatment. When these MDSCs were subsequently adoptively transferred into a murine model of renal IRI, they aggravated kidney damage, likely owing to the differentiation of M-MDSCs into deleterious macrophages and DCs.

Expression of Concern: Al-Baqami, N.M.; Hamza, R.Z. Protective Effect of Resveratrol against Hepatotoxicity of Cadmium in Male Rats: Antioxidant and Histopathological Approaches. Coatings 2021, 11, 594

Following acceptance of the Special Issue article by the Guest Editor, El-Sayed Abd El-Aziz, concerns were raised regarding the integrity of the peer review process [...]

Histopathological features of SARS-CoV-2 infection and relationships with organoid technology.

Coronavirus disease 2019 (COVID-19) following infection by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has caused a global pandemic that is still having serious effects worldwide. This virus, which targets the lungs in particular, can also damage other tissues. Angiotensin converting enzyme 2 (ACE-2) plays a key role in viral entry into host cells. The presence of ACE-2 in various tissues may permit viral infection. Studies of COVID-19 often make use of postmortem tissues. Although this information provides various useful results, it is also necessary to conduct in vitro studies to understand optimal treatment approaches. Because the virus may show species-specific differences, in vitro technologies using human cells are particularly important. Organoid technologies, three-dimensional structures that can be obtained from human cells, are playing increasingly important roles in studies of SARS-CoV-2. This technology offers a significant advantage in terms of mimicking in vivo tissue structures and testing antiviral compounds. In this mini-review, we summarize studies of SARS-CoV-2 using both histopathological and organoid technology approaches.

Poplar-type propolis provides protection of blood cells, testosterone levels and sperm motility in cisplatin-induced toxicity

Cisplatin is a frequently used chemotherapeutic in many types of cancer, especially urological cancers. Despite its efficacy in the treatment of cancer, it causes various toxic side effects such as nephrotoxicity, neurotoxicity and ototoxicity. The aim of this study was to determine the protective role of olive oil extract of propolis (OEP) with biochemical and histopathological approaches to cisplatin induced toxicity. Sixty-four adult male Wistar rats were divided into eight groups, control, propolis (100 and 200 mg/kg, orally daily for 10 days) and combined therapy (propolis 10 days before and after CP injection). Haematological (Wbc, Rbc, Mpv, Hgb, Htc, Mcv, Mch, Mchc, Lym, Neu), biochemical (triglyceride, total cholesteol, HDL and LDL-cholesterol, glucose, BUN, uric acid and testosterone level), epididymal sperm concentration, sperm motility, and histological parameters were analyzed. According to the results, cisplatin has negative effects on hematological, biochemical parameters, and sperm motility compared to control group. Administration of pre-cisplatin propolis ameliorated wbc, hct, mcv, mchc, neu levels. The administration of OEP prior to CP normalized the increased BUN and uric acid levels induced by CP. Interestingly it was also revealed that the testosterone levels increased in the OEP groups compared to the control group. Additionally, the combined administration of CP with OEP normalized the decreased testosterone levels induced by CP, particularly pre-treatment OEP. As a result, propolis, a natural product with numerous useful biological effects, was shown to a have protective as well as amelioration and normalizing effect on CP-induced damage.

Nephroprotective role of diosgenin in gentamicin-induced renal toxicity: biochemical, antioxidant, immunological and histopathological approach

BackgroundAminoglycoside antibiotics, gentamicin (GM) owns the utmost nephrotoxic potential than other antibiotics from the same category. To the other side, diosgenin (DG) showed the antioxidant and anti-inflammatory property.ResultsThe present study was aimed to explore the nephroprotective effect of diosgenin on gentamicin-induced renal toxicity in Wistar rats. Wistar albino rats were divided into six groups (n = 6): Normal control (NC), Nephrotoxicity control (GM), DG (20 mg/kg), DG (40 mg/kg), DG (80 mg/kg), accordingly. After the treatment, the nephroprotective effects of DG were assessed by measuring serum levels of creatinine (Cr), blood urea nitrogen (BUN), total proteins (TP), albumin and urea levels. Urine volume, proteins, electrolyte levels, creatinine clearance were also evaluated in urine samples. Oxidative stress was evaluated through the measurement of antioxidant stress markers in the kidney tissue. Changes in body weight and kidney weight were also recorded along with a histopathological examination of kidney sections. For evaluation of inflammation, TNF-α and IL-1β levels were measured in the blood serum using ELISA kits. GM intoxication induced elevated serum creatinine, BUN, urea, albumin and TP levels, urine electrolytes levels, pro-inflammatory cytokines, antioxidant parameters which were found to be decreased significantly in a dose-dependent manner in rat groups received DG which was also evidenced by the histological observations.ConclusionDG showed a significant nephroprotective effect in a dose-dependent manner by ameliorating the GM induced nephrotoxicity in Wistar rats.

Identification of molecular biomarkers associated with disease progression in the testis of bulls infected with Besnoitia besnoiti

Breeding bulls infected with Besnoitia besnoiti may develop sterility during either acute or chronic infection. The aim of this study was to investigate the molecular pathogenesis of B. besnoiti infection with prognosis value in bull sterility. Accordingly, five well-characterized groups of naturally and experimentally infected males were selected for the study based on clinical signs and lesions compatible with B. besnoiti infection, serological results and parasite detection. A broad panel of molecular markers representative of endothelial activation and fibrosis was investigated and complemented with a histopathological approach that included conventional histology and immunohistochemistry. The results indicated the predominance of an intense inflammatory infiltrate composed mainly of resident and recruited circulating macrophages and to a lesser extent of CD3+ cells in infected bulls. In addition, a few biomarkers were associated with acute, chronic or subclinical bovine besnoitiosis. The testicular parenchyma showed a higher number of differentially expressed genes in natural infections (acute and chronic infections) versus scrotal skin in experimental infections (subclinical infection). In subclinical infections, most genes were downregulated except for the CCL24 and CXCL2 genes, which were upregulated. In contrast, the acute phase was mainly characterized by the upregulation of IL-1α, IL-6 and TIMP1, whereas in the chronic phase, the upregulation of ICAM and the downregulation of MMP13, PLAT and IL-1α were the most relevant findings. Macrophages could be responsible for the highest level of gene regulation in the testicular parenchyma of severely affected and sterile bulls, and all these genes could be prognostic markers of sterility.

An investigation on protective effects of the new killed vaccine against nervous necrosis virus (NNV) using histopathology and immunohistochemistry approach on the brain and eye tissues of Acipenser stellatus Pallas 1771

The objective of this study was to analyze the efficiency of the killed vaccine against nervous necrosis virus on Acipenser stellutus. Heat inactivated VNN vaccine was administrated in 7 g juveniles of Acipenser stellutus as a laboratory model and it was included in three different adjuvants that were used as injection and immersion forms with different doses. Ten groups consisting of 30 A. stellutus fish in each group (group 1–4 with 3 replications, others with no replicate) were divided totally into 18 aquariums. Two steps of vaccination were done with a one-month interval and after that, all treatments and control groups were challenged by the virulent VNN virus. The mortality rate of immersion and injection groups were 12.9% and 19.8% respectively, compared to 100% mortality in the control group. Histopathology and immunohistochemistry findings were evaluated. According to the mortality rate one month after challenging, a low range mortality of 12.5% was seen in group 2 with no pathological lesion and negative IHC test in the brain and eye tissues, whereas 100% of the control group (unvaccinated group) died with severe vacuolation in the brain and eye tissues and also positive IHC test. The correlation assay between these results concluded that the immersion form with 75% of aquatic-specific Montanide IMS 1312 Seppic adjuvant made better immunization with no pathological sign or forming the complex of antigen-antibody in IHC assay. These findings are important because of the impossibility of injection in the larval stage and also due to the occurrence of the disease in the first stage of sturgeon life which could cause high mortality in susceptible fish in the larval stage.

The therapeutic effects of Bombyx mori sericin on rat skin psoriasis through modulated epidermal immunity and attenuated cell proliferation

Background and aimPsoriasis is a skin disorder that leads to chronic inflammation and keratinocyte hyperproliferation. Sericin extracted from Bombyx mori cocoon has been demonstrated to possess anti-inflammatory and antiproliferative properties, which makes it a viable candidate for psoriasis treatment. This study aimed to investigate the therapeutic effect of sericin on skin psoriasis at the cellular level. Experimental procedureImiquimod-induced skin psoriasis was established in Sprague-Dawley rats. The rats with psoriasis were divided into 6 groups (n = 5), namely, one nontreatment control group and five groups that received different treatments: sericin (2.5%, 5%, and 10%), 0.1% betamethasone, 3 μg/ml calcitriol. The treatments were administered twice daily for 7 days, followed by skin sample collection. Epidermal improvement and protein expression were evaluated using histopathological and label-free proteomic approaches and immunohistochemistry. Results and conclusionCompared with other concentrations, 10% sericin had the desired effect of improving skin psoriasis as shown by reduced epidermal thickness, similar to the effects of betamethasone and calcitriol treatments. Anti-inflammatory activity was shown by decreased C–C motif chemokine 20 (CCL20) expression posttreatment. Proteomic observation revealed that sericin reduced cytokine production by Th17 cells by interfering with the JAK-STAT signaling pathway. Sericin treatment also resulted in a modulated immune response via upregulation of Galectin-3 (Lgals3) and downregulation of Sphingosine-1-phosphate lyase1 (Sgpl1). Sericin improved epithelial cell proliferation by upregulating Nucleoside diphosphate kinase B (Nme2). Therefore, the therapeutic effect of sericin on psoriasis correlated with a reduced immune response and attenuated epidermal proliferation, making sericin a promising approach for skin psoriasis treatment.

Cerebellar spongiform degeneration is accompanied by metabolic, cellular, and motor disruption in male rats with portacaval anastomosis.

The episodes of cerebral dysfunction, known as encephalopathy, are usually coincident with liver failure. The primary metabolic marker of liver diseases is the increase in blood ammonium, which promotes neuronal damage. In the present project, we used an experimental model of hepatic encephalopathy in male rats by portacaval anastomosis (PCA) surgery. Sham rats had a false operation. After 13weeks of surgery, the most distinctive finding was vacuolar/spongiform neurodegeneration exclusively in the molecular layer of the cerebellum. This cerebellar damage was further characterized by metabolic, histopathological, and behavioral approaches. The results were as follows: (a) Cellular alterations, namely loss of Purkinje cells, morphological changes, such as swelling of astrocytes and Bergmann glia, and activation of microglia; (b) Cytotoxic edema, shown by an increase in aquaporin-4 and N-acetylaspartate and a reduction in taurine and choline-derivate osmolytes; (c) Metabolic adjustments, noted by the elevation of circulating ammonium, enhanced presence of glutamine synthetase, and increase in glutamine and creatine/phosphocreatine; (d) Inflammasome activation, detected by the elevation of the marker NLRP3 and microglial activation; (e) Locomotor deficits in PCA rats as assessed by the Rotarod and open field tests. These results lead us to suggest that metabolic disturbances associated with PCA can generate the cerebellar damage that is similar to morphophysiological modifications observed in amyloidogenic disorders. In conclusion, we have characterized a distinctive cerebellar multi-disruption accompanied by high levels of ammonium and associated with spongiform neurodegeneration in a model of hepatic hypofunctioning.

Outbreak investigation attributes Infectious spleen and kidney necrosis virus as a necessary cause of a mortality epidemic in farmed grouper (Epinephelus spp.) in Bali, Indonesia

In 2016, famers reported mass mortality events in hybrid grouper within 2–4 weeks following transfer to sea cages on the northern coast of Bali. The objective was to obtain an aetiological diagnosis using a broad range of traditional and emerging diagnostic approaches. A group of 24 and 12 fish with and without clinical signs were sampled from 10 affected populations at 9 farms. Samples for histopathology, ectoparasites evaluation and molecular approaches for microbiology were obtained with a diagnostic post-mortem examination. Fish with clinical signs had a significantly higher likelihood of having pale anterior and posterior kidneys and a liver that was pale and reduced in size compared to fish without clinical signs. There were no differences in the prevalence and quantity of Megalocytivus (MCV) or nervous necrosis virus (NNV) in tissues observed from fish with and without clinical signs. Nearly 55 % of fish were infected with NNV irrespective of clinical signs. There were no histopathological lesions consistent with virial nervous necrosis and the NNV infections were considered subclinical. 80 % of grouper were infected with MCV, irrespective of clinical signs. A significant proportion of fish with clinical signs (true prevalence 94.4 %; 95 % CI 79–100) had observed megalocytes and pathology consistent with disease caused by ISKNV compared to those without clinical signs (true prevalence 47.2 %; 95 % CI 27–70). Metagenomic sequences generated using Illumina Miseq and taxonomically labelled using BlastN + revealed that the Megalocytivirus was 99.9 % similar to Infectious spleen and kidney necrosis virus (ISKNV). The unbiased sequencing did not detect any novel DNA viruses or bacterial pathogens of clinical significance. The monogenean, Benedenia epinepheli and a leech, Zeylanicobdela arugamensis were detected at 6/9 and 9/9 farms, respectively. Our approach identified several pathogens reported in grouper aquaculture with histopathology showing that ISKNV was a necessary cause for the mass mortality events.

Real-Time Imaging of Hepatic Inflammation Using Hydrogen Sulfide-Activatable Second Near-Infrared Luminescent Nanoprobes

The sensing and visualized monitoring of hydrogen sulfide (H2S) in vivo is crucial to understand its physiological and pathological roles in human health and diseases. Common methods for H2S detection require the destruction of the biosamples and are not suitable to be applied in vivo. In this Communication, we report a "turn-on" second near-infrared (NIR-II) luminescent approach for sensitive, real-time, and in situ H2S detection, which is based on the absorption competition between the H2S-responsive chromophores (compound 1) and the NIR-II luminescent lanthanide nanoparticles. Specifically, the luminescence was suppressed by compound 1 due to the competitive absorption of the incident light. In the presence of H2S, the compound 1 was bleached to recover the luminescence. Thanks to the deep tissue penetration depth and the low absorbance/scattering on biological samples of the NIR-II nanoprobes, the monitoring of the endogenous H2S in lipopolysaccharide-induced liver inflammation was achieved, which is unattainable by the conventional histopathological and serological approaches.

The novel approach to distinguish primary multiple lung adenocarcinomas from intrapulmonary metastases by next generation sequencing in Chinese patients.

e20506 Background: Differentiation of multiple primary lung cancer (MPLC) from intrapulmonary metastasis (IPM) is critical to determine clinical stage. Although clinicopathological features could provide certain evidences, it’s still challenging to identify the tumor malignancy accurately. In General, standard histopathologic approach is adequate in most cases, but has notable limitations in the recognition of IPMs. Herein, we propose an integrated molecular algorithm to facilitate MPLCs and IPMs diagnosis in the clinical practice. Methods: 40 Chinese patients with lung adenocarcinomas were enrolled in the study, 84 tumor samples were collected for next-generation sequencing. Somatic alterations with variant allele fraction (VAF) ≥1% were taken into account for molecular algorithm. A genomic database of 2,471 Chinese lung adenocarcinomas (LUAD) was used to calculate odds of coincidental occurrence, prevalence of individual mutation prevalence. Tumor relatedness diagnosed by histopathologic assessment was contrasted with comparative genomic profiling by subsequent NGS. Moreover, the performance of molecular algorithm prediction was evaluated as well. Results: Firstly, we compared the performance of comprehensive next-generation sequencing (NGS) with standard histopathologic approaches for distinguishing NSCLC subtypes in clinical practice. The genomic profiling was described as following: EGFR alterations occurred more frequently in MPLCs compared to IPMs (77.1% vs 50.0%, P<0.05). Further analysis showed that TP53 alterations occurred less frequently in MPLCs compared to large Chinese cohort (22.9% vs 51.0%, P<0.05). TP53 alterations occurred less frequently in MPLCs compared to large Chinese cohort (P<0.05). The classifications based on the three different methodologies mentioned above were compared. Molecular algorithm prediction was concordant with NGS in 21 cases (52.5%), particularly in the prediction of MPLC. Retrospective review highlighted several histologic challenges, including morphologic progression in some IPMs. For the five undetermined cases, two showed differences in architectural patterns, and remained cases have nodules presented as adenocarcinoma in situ, or minimally invasive adenocarcinoma. Of 28 MPLC cases defined by NGS, 25 cases had unique somatic mutations per pair Based on calculation from the prevalence of EGFR L858R mutation (27%) in large Chinese cohort, the odds of coincidental occurrence of the mutation in two unrelated tumors was 7.3%. Taking together, EGFR alterations occurred more frequently in MPLCs compared to IPMs (77.1% vs 50.0%, P<0.05). Molecular algorithm prediction was concordant with NGS in 21 cases (52.5%). Conclusions: Our results support broad panel NGS to assist differential diagnosis to assist approach in clinical practice. It is necessary to conduct large clinical study to establish comprehensive algorithm models to assist diagnosis and predict clinical outcome.

Protective Effect of Resveratrol against Hepatotoxicity of Cadmium in Male Rats: Antioxidant and Histopathological Approaches

Cadmium (Cd) is widely used in some industries and emitted from fossil fuels. It is a heavy metal with a number of side effects, including hepatotoxicity. Resveratrol (Rs) is considered an important polyphenol, which is a secondary plant metabolite and has the ability to scavenge free radicals. The study was designed to evaluate the effects of resveratrol on Cd, which induced hepatotoxicity, by the assessment of some histopathological and biochemical alterations. Forty male albino rats were divided into four groups: the 1st group was the control group, the 2nd group was treated with Cd (5 mg/kg), the 3rd group was given Rs (20 mg/kg), and the 4th group was treated with Cd in combination with Rs intraperitoneally for 30 successive days. The results indicate that Cd increased liver enzymes alanine aminotransferase and aspartate aminotransferase (AST and ALT), alkaline phosphatase ALP and gamma-glutamyl transferase (γ-GT) while reducing the total protein level; Cd increased the malondialdhyde (MDA) level while decreasing the levels of other antioxidant enzymes super oxide dismutase, catalase and glutathione peroxidase (SOD, CAT and GPx). Serious congestion and hemorrhage related to the hepatic tissues were noticed in the Cd group, and Rs plays a major role in alleviating histopathological injuries and hepatic oxidative damage. It is clear that Rs has the ability to minimize the hepatotoxicity induced by Cd in male rats.

Routine Histopathological Examination of the Specimen After Laparoscopic Cholecystectomy: Can We Be Brave Enough to Give Up?

Selective versus routine histopathological examination after cholecystectomy is still in debate. This study aims to investigate the effect of histopathology results on treatment modality and surgery strategy. The validity of the selective histopathology approach was questioned. The data of patients undergoing laparoscopic cholecystectomy between January 2009 and December 2019 were retrospectively analyzed. The demographics and histopathology results, whether the operation was emergent or elective, and the reasons for conversion to open surgery were recorded. Malignant and precursor histopathology diagnoses were examined, and their relationship with the surgical strategy was questioned. A total of 2723 patients were included in the study. Of these patients, 2600 (95.5%) were operated under elective, while 123 (4.5%) were operated under emergency conditions. While the surgery was completed laparoscopically in 2685 (98.6%) patients, it was converted to open surgery in 38 (1.4%) patients. Age, gender, the presence of primary gallbladder cancer, acute cholecystitis, and xanthogranulomatous cholecystitis in histopathological examination were found to be independent predictive factors for conversion to open surgery (P < .05). The rate of primary invasive carcinoma in the series was 0.1%. Routine histopathological examination of the gallbladder is important for demonstrating a wide spectrum of pathological changes in this organ. Invasive cancer or precursor lesions can be detected even in patients without any macroscopic abnormality. Histopathological examination also plays a role in determining follow-up, further examination, and treatment modality in addition to the diagnosis in these patients.

Effects of tamoxifen therapy on sciatic nerve crush injury: An experimental study in rats.

This study aimed to determine the effects of tamoxifen on sciatic nerve crush injury in a rat model using histopathological, histomorphometric, and immunohistochemical approaches. In this study, 24 male Sprague-Dawley rats aged of 5 to 7 weeks and weighing between 300 g and 400 g were used. The rats were randomly divided into 3 groups: control (group C), sciatic nerve injury (group SNI), and sciatic nerve injury with tamoxifen (group SNT). The sciatic nerve crush injury model was performed using the De Koning's crush force method. In group C, only a skin incision was made and then the skin was sutured. In group SNI, the injury model was performed but no treatment was applied. In group SNT, the injury model was executed, and then 40 mg/kg/day tamoxifen was given for 4 weeks by intraperitoneal methods. At the end of 4 weeks, all animals were killed using high doses of an anesthetic. Approximately, 2-cm sciatic nerve samples were obtained for histopathological, histomorphometric, and immunohistochemical analyses using the old skin incision. In the histopathological examination, vascular congestion and density of vacuolization were significantly lower in group SNT than in group SNI (p<0.05). In the histomorphometric examination, the mean sciatic nerve diameter was 306±62 µm in group C, 510±42 µm in group SNI, and 204±23 µm in group SNT. A significant difference was observed in the sciatic nerve diameter measurements among the 3 groups (p<0.05). In pairwise comparisons, the mean sciatic nerve diameter was significantly lower in group SNT than in group SNI (p=0.00002). Sciatic nerve diameter measurements of both groups were found to be significantly higher than group C (p<0.05). The mean epineurium thickness was 17±0.8 µm in group C, 32±2.5 µm in group SNI, and 17±0.8 µm in group SNT. A significant difference was observed in the epineurium thickness measurements among the 3 groups (p<0.05). In pairwise comparisons, the epineurium thickness was significantly higher in group SNI than in groups SNT and C (p<0.05). In the immunohistochemical analysis, S100 immunoreactivity was found significantly higher in group SNI than in the other 2 groups (p<0.05). The histomorphometric, histopathological, and immunohistochemical data obtained from this study have shown that tamoxifen has a beneficial effect on sciatic nerve crush injury in the experimental rat model.

Chia Seed Oil Ameliorates Doxorubicin-Induced Cardiotoxicity in Female Wistar Rats: An Electrocardiographic, Biochemical and Histopathological Approach

Doxorubicin (DOX) is a potent anti-cancer antibiotic that was widely used for treatment of various cancers. It produces free radicals which result in extreme dose-limiting cardiotoxicity. This study investigated the cardioprotective potential of chia seed oil, an active polyphenolic nutraceutical against doxorubicin-induced cardiotoxicity in Wistar rats. Twenty-four female Wistar rats were divided into four groups (n = 6) which consist of normal control, DOX control, test-A and test-B group. Animals were prophylactically treated with two different doses of test drug, i.e. chia seed oil 2.5 ml/kg/day and 5 ml/kg/day in test-A and test-B groups orally for 7 days. Doxorubicin (25 mg/kg; single dose) was administered intraperitoneally to DOX control, Test-A and Test-B animals on the seventh day to induce cardiotoxicity. ECG analysis was done before and after treatment. Besides ECG, CK, CK-MB, LDH, AST, MDA and GSH were analyzed. DOX had significantly altered ECG, CK, CK-MB, LDH, AST, MDA and GSH. Pre-treatment with chia seed oil significantly alleviated DOX-induced ECG changes and also guarded against DOX-induced rise of serum CK, CK-MB and AST levels. Chia seed oil alleviated histopathological alteration in DOX-treated rats. It also significantly inhibited DOX-induced GSH depletion and elevation of MDA. The present study revealed that chia seed oil exerts cardioprotection against doxorubicin-induced cardiotoxicity in female Wistar rats. Our study opens the perspective to clinical studies to precisely consider chia seed oil as a potential chemoprotectant nutraceutical in the combination chemotherapy with doxorubicin to limit its cardiotoxicity.

CD11c-CD8 Spatial Cross Presentation: A Novel Approach to Link Immune Surveillance and Patient Survival in Soft Tissue Sarcoma.

Simple SummaryImmune cells can be powerful regulators of tumor growth and disease progression. Yet, the potential role of professional antigen-presenting cells in soft tissue sarcoma is poorly explored. Both dendritic cells and macrophages may present exogenous antigens through major histocompatibility complex (MHC) class I molecules to CD8+ T cells, a process referred to as cross presentation. With the concept of cellular cross presentation in mind, the present study supports the hypothesis that CD11c+ cells in direct cell-cell contact with CD8+ T cells within the primary tumor are associated with an active anti-tumor immune microenvironment and favorable prognosis. Our work hereby defines a novel biomarker for immune surveillance where presence of spatial cross presentation at tissue level resolution is significantly associated with overall survival. Importantly, this biomarker is independent from established prognostic markers like tumor grade. Biomarkers linked to immune surveillance are expected to gain increasing attention with the advent of immunotherapy in clinical practice.Checkpoint inhibitors are slowly being introduced in the care of specific sarcoma subtypes such as undifferentiated pleomorphic sarcoma, alveolar soft part sarcoma, and angiosarcoma even though formal indication is lacking. Proper biomarkers to unravel potential immune reactivity in the tumor microenvironment are therefore expected to be highly warranted. In this study, intratumoral spatial cross presentation was investigated as a novel concept where immune cell composition in the tumor microenvironment was suggested to act as a proxy for immune surveillance. Double immunohistochemistry revealed a prognostic role of direct spatial interactions between CD11c+ antigen-presenting cells (APCs) and CD8+ cells in contrast to each marker alone in a soft tissue sarcoma (STS) cohort of 177 patients from the Karolinska University Hospital (MFS p = 0.048, OS p = 0.025). The survival benefit was verified in multivariable analysis (MFS p = 0.012, OS p = 0.004). Transcriptomics performed in the TCGA sarcoma cohort confirmed the prognostic value of combining CD11c with CD8 (259 patients, p = 0.005), irrespective of FOXP3 levels and in a CD274 (PD-LI)-rich tumor microenvironment. Altogether, this study presents a histopathological approach to link immune surveillance and patient survival in STS. Notably, spatial cross presentation as a prognostic marker is distinct from therapy response-predictive biomarkers such as immune checkpoint molecules of the PD-L1/PD1 pathway.

A multidisciplinary approach for the differential diagnosis between multiple primary lung adenocarcinomas and intrapulmonary metastases

PurposeThe distinction between multiple primary lung cancers (MPLCs) and intrapulmonary metastases has a significant impact on tumor staging and therapeutic choices. Several criteria have been proposed to solve this diagnostic issue, but a definitive consensus is still missing. We tested the efficacy of a combined clinical, histopathological and molecular (“real world”) approach for the correct classification of multiple lung tumors in a selected cohort of patients. Methods24 multiple lung tumors with a diagnosis of adenocarcinoma from 10 patients were retrospectively reviewed. Radiological, pathological and clinical information, including follow-up, were integrated with molecular profiling via a routine multigene panel sequencing. ResultsComprehensive histologic assessment revealed readily distinguishable histologic patterns between multiple tumors suggesting unrelated lesions in 7 cases, in agreement with clinical, radiological and molecular data, thus leading to final diagnosis of MPLCs. In the remaining 3 cases, the differential diagnosis between MPLCs and intrapulmonary metastases was challenging, since the histologic features of the lesions were similar or identical. The final interpretation (2 MPLCs and 1 most likely intrapulmonary metastases) was reached thanks to the integration of all available data, and was confirmed by follow-up. ConclusionsA multidisciplinary approach including a routinely affordable multigene panel sequencing is a useful tool to discriminate MPLCs from intrapulmonary metastases in multiple lung nodules sharing the adenocarcinoma histotype.