Immunohistological Examination Research Articles

Influence of mechanical stimuli and zoledronic acid on the femoral morfology in rats with obesity and limited mobility models

The aim. Investigate the long-term effect of WBV and bisphosphonate (zoledronic acid) on the morphological structure and mineral density of the rat femora. Materials and methods. Rats were divided into control and three experimental groups: I - experimental group - high-calorie diet + limited mobility (HCD + LM), II - experimental group - HCD + LM + whole body vibration (WBV 3 m/s2). III - experimental group - HCD + LM + administration of zoledronic acid (HCD + LM + Zol. at a dose of 0.025 mg / kg). Bone mineral density determination, scanning electron microscopy of trabecular meshwork and immunohistological examination were performed. Results. The application of the model of a high-calorie diet with limited mobility leads to the loss of the mineral component of trabeculae up to 16.2%. The trabecular layer thinned to 13.9% compared to the control group, and the intertrabecular spaces expanded. Under the influence of whole-body vibration, there was an increase in BMD in the femora compared to the group HCD + LM, and bone density remained at the control level until the 16th week of the experiment. In group III, the rate of bone formation increased, the ratio of bone to trabecular volume was significantly higher compared to the group HCD + LM. Starting from the 8th week, it increased by 8.1% and 13% at the end of the experiment, respectively.Conclusions. A high-calorie diet with limited mobility reduces the mineral density of the trabeculae in the femoral bone. Zoledronic acid effectively improves mineralization but leads to impairment of the morphology of the bone growth plate. The research results indicate that WBV can be an effective non-pharmacological method of maintaining bone health in young people.

Ultra-High Dose Rate FLASH Effect on Head and Neck Cancer Cells and Normal Salivary Gland in Mice

<h3>Purpose/Objective(s)</h3> Ultra-high dose rate irradiation (FLASH) is a novel therapeutic modality that has been reported to spare tissues compared to conventional dose rate. We aimed to investigate the effect of FLASH on head and neck (H&N) cancer cells and salivary gland dysfunction caused by radiotherapy. <h3>Materials/Methods</h3> H&N cancer cells (HSG and FaDu) were irradiated with 10 MeV electron beam at a dose rate of 132.2Gy/s (FLASH) or 0.1Gy/s (CONV), by a modified clinic linear accelerator. The irradiated dose was confirmed with EBT-XD film, and 3, 7, and 11 Gy were irradiated. We evaluated the cell cycle and the surviving fraction of cells with clonogenic assays. We administered single dose head and neck irradiation to mice with a 6 MeV electron beam at dose rate of 105.6-126 Gy/s (FLASH) or 0.03Gy/s (CONV), using a modified linear accelerator. The irradiated dose was confirmed with EBT3 film, and 9-76 Gy and 10-30 Gy were irradiated for FLASH and CONV, respectively. Parotid and submandibular glands were obtained from mice at 3 and 30 days after irradiation. Immunohistological examination and apoptotic assays were conducted to assess the radiation response. <h3>Results</h3> The proportion of sub G0/G1 was higher in the FLASH group, and the surviving fraction was significantly decreased in FLASH through every dose range. At three days post-irradiation, both 10 Gy and 30 Gy irradiated groups showed a greater number of salivary cells and increased expression of AQP5 in FLASH compared to conventional irradiated mice. As a late toxicity (30 days of irradiation), fibrosis did not show any significant difference, although FLASH group received much higher radiation dose. There was no difference in apoptosis between groups on post 3 days, but FLASH showed significantly less degree of apoptosis on 30 days after irradiation. <h3>Conclusion</h3> This study demonstrates that FLASH irradiation strengthens the therapeutic effect for H&N cancers and attenuates radiation-induced salivary gland damage. Further research is needed on the biological mechanisms involved.

Effect of crude polysaccharide from seaweed, Dictyopteris divaricata (CDDP) on gut microbiota restoration and anti-diabetic activity in streptozotocin (STZ)-induced T1DM mice

Type-1 Diabetes Mellitus (T1DM) is regarded as a multifunctional, immune-related disease which causes massive destruction of islet β-cells in pancreas resulting in hyperglycemic, hypoinsulinemia and hyperlipidimic conditions. The aim of the present study, was to investigate the hypothesis that streptozotocin (STZ)-induced T1DM in Balb/c mice when treated with crude polysaccharide from seaweed, Dictyopteris divaricata (CDDP) depicts improvement in diabetes-related symptoms. Treatment with CDDP resulted in decreased body weight loss, improved food consumption and water intake disbalances. The CDDP effectively improved fasting blood glucose, oral glucose tolerance (OGTT), serum insulin, insulin secretion, rejuvenation of β-cells mass, serum lipid profile and pro-inflammatory cytokines levels. Additionally, treatment with CDDP increased the population of beneficial bacteria such as Firmicutes, Bacteroidetes and Lactobacillus at phylum, family and genus levels by 16S rRNA sequencing. Furthermore, immunohistological examination confirmed that CDDP reduces the inflammation and restored the structural morphology of colon and upraised the levels of insulin receptor substrate-1 (IRS-1), Mucin-2 (MUC-2) and tight-junction proteins (TJs) whereby maintaining the gut structures and barrier permeability. Thus, the above presented data, highlights the safe and therapeutic effects of crude polysaccharide (CDDP) from D. divaricata in the treatment and restoration of T1DM disorders and can be used as a food supplement alternative to diabetes medicine.

I-FABP as a Potential Marker for Intestinal Barrier Loss in Porcine Polytrauma.

Polytrauma and concomitant hemorrhagic shock can lead to intestinal damage and subsequent multiple organ dysfunction syndrome. The intestinal fatty acid-binding protein (I-FABP) is expressed in the intestine and appears quickly in the circulation after intestinal epithelial cell damage. This porcine animal study investigates the I-FABP dynamics in plasma and urine after polytrauma. Furthermore, it evaluates to what extent I-FABP can also act as a marker of intestinal damage in a porcine polytrauma model. Eight pigs (Sus scrofa) were subjected to polytrauma which consisted of lung contusion, tibial fracture, liver laceration, and hemorrhagic shock followed by blood and fluid resuscitation and fracture fixation with an external fixator. Eight sham animals were identically instrumented but not injured. Afterwards, intensive care treatment including mechanical ventilation for 72 h followed. I-FABP levels in blood and urine were determined by ELISA. In addition, immunohistological staining for I-FABP, active caspase-3 and myeloperoxidase were performed after 72 h. Plasma and urine I-FABP levels were significantly increased shortly after trauma. I-FABP expression in intestinal tissue showed significantly lower expression in polytraumatized animals vs. sham. Caspase-3 and myeloperoxidase expression in the immunohistological examination were significantly higher in the jejunum and ileum of polytraumatized animals compared to sham animals. This study confirms a loss of intestinal barrier after polytrauma which is indicated by increased I-FABP levels in plasma and urine as well as decreased I-FABP levels in immunohistological staining of the intestine.

Oral Focal Mucinosis, an uncommon entity: Report of three cases

<h3>Introduction</h3> Oral focal mucinosis (OFM) is a rare soft tissue lesion of unknown etiology and it is considered the oral counterpart of cutaneous focal mucinosis or cutaneous myxoid cyst. The pathogenesis of OFM may be explained by fibroblast overproduction of hyaluronic acid. Clinically, it appears as painless swelling of the same color as the surrounding mucosa. The gingiva is the most common site and the diagnosis is established only by histopathological and immunohistological examination. <h3>Materials and Methods</h3> We present three cases of gingival oral focal mucinosis. Case 1: 40 year-old womatn with painless and slow-growing swelling of two years of evolution at buccal mandibular gingiva, between lateral incisor and canine. The lesion measured 2 cm. and exhibited the same color as the overlying mucosa; there was not radiographic involvement. Case 2: 48 year-old female with a hard palatal swelling. The lesion was sessile, firm, and not color change Case 3: 36 year-old male with a painless and same color as the normal mucosa gingival swelling. The lesion was between central and lateral right incisors, it was sessil and measured 1.5 cm Results In the three cases excisional biopsy was performed and histopathologically they shown the proliferation of elongated and mucinous material in a myxoid matrix. This myxoid stroma was surrounded by collagenous fibrous connective tissue and covered with squamous epithelium. Histochemical staining with Alcian blue was positive in the mucinous areas Conclusions These three cases are significant clinically as well as histopathologically. The lesions clinically mimic an ossifying peripheral fibroma or a fibroma. Thus, the histopathological and histochemical findings with special stains, such as Alcian blue, are determinant to establish the diagnosis of these conditions. Treatment is the complete surgical excision and its recurrence is not reported.

Primary Extra Nodal Diffuse Large B-Cell Lymphoma of the Maxillary Sinus with Symptoms of Acute Pulpitis

Diffuse large B-cell lymphoma not otherwise specified (DLBCL-NOS) is a subtype of large B-cell non-Hodgkin lymphoma with various clinical and pathological manifestations. DLBCL-NOS which primarily arises from maxillary sinus is rare and hard to diagnose due to unique anatomy. Here, we present a case of DLBCL-NOS that developed in the left maxillary sinus of a 72-year-old male, who presented with severe toothache that resembled acute pulpitis. The lesion was diagnosed and treated based on radiographs, histological, immunohistological examinations, and PET-CT analysis. Despite its rare incidence, DLBCL-NOS should still be included in differential diagnoses to rule out malignancy in cases of endodontic disease.

Epidermolysis Bullosa With Congenital Absence of Skin: Congenital Corneal Cloudiness and Esophagogastric Obstruction Including Extended Genotypic Spectrum of PLEC, LAMC2, ITGB4 and COL7A1.

Epidermolysis bullosa (EB) is a rare and genetically heterogeneous disorder characterized by skin fragility and blister formation occurring spontaneously or after minor trauma. EB is accompanied by congenital absence of skin (EB with CAS) in some patients. Pathogenic variants of COL7A1 are responsible for EB with CAS in the vast majority of cases. Type and subtype diagnosis of EB with CAS generally requires specific immunohistological examinations that are not widely available plus targeted gene analysis. The present study aimed to determine the clinical features of five patients affected by EB with CAS and to identify the underlying genetic defects using whole exome sequencing (WES) followed by focused analysis of the target genes. Four patients had generalized skin involvement and one had localized defects. Two patients exhibited extremely severe skin manifestations and congenital cloudy cornea along with pyloric atresia, and one had partial esophagogastric obstruction and anuria due to vesicoureteric obstruction. In the WES analysis, the average coverage of the target exons was 99.05% (726 of 733 exons), with a range of 96.4–100% for individual genes. We identified four novel and two known pathogenic/likely pathogenic variants of five distinct genes in the examined families: PLEC:c.2536G > T (p.Glu846Ter); LAMC2:c.3385C > T (p.Arg1129Ter); KRT5:c.429G > A (p.Glu477Lys); ITGB4:c.794dupC (p.Ala266SerfsTer5); COL7A1:c.5440C > T (p.Arg1814Cys); and COL7A1:c.6103delG. All alleles were inherited from the parents, except for the KRT5 variant as a de novo finding. The findings reveal extremely rare phenotypes found in EB with CAS, namely congenital cloudy cornea, esophagogastric obstruction, and anuria, and extend the genotypic spectrum of EB-related genes. The data confirm that WES provides very high coverage of coding exons/genes and support its use as a reasonable alternative method for diagnosis of EB. The present data from an underrepresented population in Southeast Asia could further broaden the knowledge and research on EB.

Metastatic papillary thyroid carcinoma presenting with elevated serum levels of carbohydrate antigen 19-9 (CA19-9): a case report

BackgroundThe major sites of distant metastases of papillary thyroid carcinoma (PTC) are the lung and bone; metastasis to the liver is rare. Although the postoperative serum thyroglobulin (Tg) level after total thyroidectomy is a good prognostic indicator for PTC when anti-thyroglobulin antibody (TgAb) is negative, the presence of TgAb interferes with the Tg assay, making serum Tg levels unreliable. Here we report a case of liver metastasis of PTC that presented with elevated serum levels of carbohydrate antigen 19-9 (CA19-9), which is usually a serum marker of pancreatic and gastrointestinal neoplasias.Case presentationA 69-year-old man was diagnosed with PTC and underwent total thyroidectomy 16 years ago. The patient’s serum Tg levels increased progressively during follow-up and his serum TgAb was negative. Positron emission tomography (PET) and computed tomography (CT) revealed metastases of the lung, cervical spine, mediastinum and liver. The liver lesion was a solitary tumor measuring 4.0 cm in the greatest dimension. His serum CA19-9 level was very high (326 U/mL), and intrahepatic cholangiocarcinoma was suspected from the results of various examinations including gastrointestinal endoscopic imaging and CT. Laparoscopic partial liver resection for segment 4 was performed. The histopathological diagnosis was a metastatic liver tumor from PTC. The immunohistological examination revealed that the liver tumor was positive for CA19-9 and Tg. The primary PTC, recovered from paraffin-embedded specimen, was also positive for CA19-9. After the surgery, his serum CA19-9 level as well as serum Tg level markedly decreased.ConclusionsWe presented the first reported case of liver metastasis of a PTC presenting with elevated serum levels of CA19-9 after total thyroidectomy. This case suggests that the serum CA19-9 levels may serve as a surrogate marker for PTC in place of the serum Tg level in patients with positive serum TgAb if the PTC and/or the metastatic lesions are positive for CA19-9 staining.

Ex Vivo Fluorescence Confocal Microscopy in Specimens of the Liver: A Proof-of-Concept Study.

Simple SummaryFluorescence confocal microscopy (FCM) is a novel micro-imaging technique providing optical sections of native tissue. The method is increasingly used for the routine diagnosis of tumors and inflammatory lesions of the skin and shows promising results for the diagnosis of other organ tumors. Very few publications exist about examinations of liver tissue thus far. In this study, we compare findings of FCM-examinations of biopsies and surgical specimens of the liver with the final diagnoses from conventional histology.Ex vivo Fluorescence Confocal Microscopy (FCM) is a technique providing high-resolution images of native tissues. The method is increasingly used in surgical settings in areas of dermatology and urology. Only a few publications exist about examinations of tumors and non-neoplastic lesions of the liver. We report on the application of FCM in biopsies, surgical specimens and autopsy material (33 patients, 39 specimens) of the liver and compare the results to conventional histology. Our preliminary examinations indicated a perfect suitability for tumor diagnosis (ĸ = 1.00) and moderate/good suitability for the assessment of inflammation (ĸ = 0.4–0.6) with regard to their severity and localization. Macro-vesicular steatosis was reliably detected, micro-vesicular steatosis tended to be underestimated. Cholestasis and eosinophilic granules in granulocytes were not represented in the scans. The tissue was preserved as native material and maintained its quality for downstream histological, immunohistological and molecular examinations. In summary, FCM is a material sparing method that provides rapid feedback to the clinician about the presence of tumor, the degree of inflammation and structural changes. This can lead to faster therapeutic decisions in the management of liver tumors, treatment of hepatitis or in liver transplant medicine.

Neuroprotective Effect of Glatiramer Acetate on Neurofilament Light Chain Leakage and Glutamate Excess in an Animal Model of Multiple Sclerosis.

Axonal and neuronal pathologies are a central constituent of multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE), induced by the myelin oligodendrocyte glycoprotein (MOG) 35–55 peptide. In this study, we investigated neurodegenerative manifestations in chronic MOG 35–55 induced EAE and the effect of glatiramer acetate (GA) treatment on these manifestations. We report that the neuronal loss seen in this model is not attributed to apoptotic neuronal cell death. In EAE-affected mice, axonal damage prevails from the early disease phase, as revealed by analysis of neurofilament light (NFL) leakage into the sera along the disease duration, as well as by immunohistological examination. Elevation of interstitial glutamate concentrations measured in the cerebrospinal fluid (CSF) implies that glutamate excess plays a role in the damage processes inflicted by this disease. GA applied as a therapeutic regimen to mice with apparent clinical symptoms significantly reduces the pathological manifestations, namely apoptotic cell death, NFL leakage, histological tissue damage, and glutamate excess, thus corroborating the neuroprotective consequences of this treatment.

Neuropathology of SCA34 showing widespread oligodendroglial pathology with vacuolar white matter degeneration: a case study

Spinocerebellar ataxia type 34 (SCA34) is an autosomal dominant inherited ataxia due to mutations in ELOVL4, which encodes one of the very long-chain fatty acid elongases. SCA38, another spinocerebellar ataxia, is caused by mutations in ELOVL5, a gene encoding another elongase. However, there have been no previous studies describing the neuropathology of either SCA34 or 38. This report describes the neuropathological findings of an 83-year-old man with SCA34 carrying a pathological ELOVL4 mutation (NM_022726, c.736T>G, p.W246G). Macroscopic findings include atrophies in the pontine base, cerebellum, and cerebral cortices. Microscopically, marked neuronal and pontocerebellar fiber loss was observed in the pontine base. In addition, in the pontine base, accumulation of CD68-positive macrophages laden with periodic acid-Schiff (PAS)-positive material was observed. Many vacuolar lesions were found in the white matter of the cerebral hemispheres and, to a lesser extent, in the brainstem and spinal cord white matter. Immunohistological examination and ultrastructural observations with an electron microscope suggest that these vacuolar lesions are remnants of degenerated oligodendrocytes. Electron microscopy also revealed myelin sheath destruction. Unexpectedly, aggregation of the four-repeat tau was observed in a spatial pattern reminiscent of progressive supranuclear palsy. The tau lesions included glial fibrillary tangles resembling tuft-shaped astrocytes and neurofibrillary tangles and pretangles. This is the first report to illustrate that a heterozygous missense mutation in ELOVL4 leads to neuronal loss accompanied by macrophages laden with PAS-positive material in the pontine base and oligodendroglial degeneration leading to widespread vacuoles in the white matter in SCA34.

Receptor-related aspects of reduced lubrication in women of reproductive age with hypoandrogenism

Aim — to investigate receptivity of the vaginal epithelium in women with the female sexual dysfunction and hypoandrogenism.Materials and methods. The immunohistochemical investigation has been performed to define the density of androgen receptors (AR) and estrogen receptors (ER) in the distal part of the vaginal tract of women of reproductive age with hypoandrogenic provision of their hormonal background. For immunohistological examination, a biopsy of the lower third of the vaginal mucosa was taken using a dermopunch with a diameter of 3 mm in 20 women with female sexual dysfunction (FSD) and hypo­andro­genism. The control group consisted of 20 women of reproductive age without FSDmanifestations and disturbances in their hormonal levels. The survey was carried out at the stage of pre-conceptional preparation. Location: Ukrainian Scientific and Prac­tical Center for Endocrine Surgery, Transplantation of Endocrine Organs and Tissues of the Ministry of Health of Ukraine.Results. Immunoreactivity of ERα in the epithelium in the main and control groupswas 47.2 ± 22.2 and 86.1 ± 8.0 (p &lt; 0.001), in the stroma — respectively 13.9 ± 16.8 and 86.1 ± 8.3 (p &lt; 0.001), that is, immunoreactivity in the epithelium was significantly higher thanin the stroma (p &lt; 0,01). The ERα locationin the vaginal epithelium was the samein both groups: cellsof the basal-intermediate layer along the basement membrane and the intermediate layer. No cells, expressing ERα, were detected in the surface layer. In stromal cells, ERαcontained in fibroblasts and smooth muscle cells. ERβ was found in cellsof thebasal — parabasal layer, the intermediate layer, occasionally —on the surface layerof vaginal epithelium, as well as in fibroblasts, vascular endothelium, where both nuclear andcytoplasmic staining was revealed. According to the results of immunohistochemical study of AR invagina, the proportion of AR-positive cells in healthy women was negligible (3.8 ± 2.2), that is considerably less than in patients with androgen deficiency (18.8 ± 8.3) and compared withother receptors. AR were localized mainly in the basal layer alongbasement membrane. Single AR-positive fibroblasts occurred in the stroma.Conclusions. Estrogen receptors α (ERα) are involved in the regulation of the processes of proliferation and differentiation of the vaginal epithelium under the influence of estrogen, as evidenced by a 1.8-fold increase in the density of stained ERα in women of the control group. The absence of correlations between ЕRα and ЕRβ receptors (r = 0.24; p &lt; 0.05) may indicate the ability to change in the same tissue depending on the strength of estrogen effects on the body of a woman. Topical localization of androgen receptors in the basal layer testifies in favour of the safe use of local formsof androgensto overcome the reduced lubrication.

Ameliorative impact of taurine on oxidative damage induced by Ipomoea carnea toxicity in wistar male rats through modulation of oxidative stress markers, apoptotic and Nrf2 pathway

ObjectivesThe adverse deleterious influences of Ipomoea carnea on brain tissue and Taurine's relative ability to prevent this neurotoxicity have been examined. MethodsWe utilized sixty Wistar male rats weighing 1752 g. The rats were allotted into 4 groups of fifteen each: The control group got saline i.p. daily for eight weeks; the Ipomoea carnea extract (ICE) group received ICE (15 mg/kg b.w./day) orally for eight weeks. Taurine-treated rats, in which rats were administered Taurine (200 mg·kg−1, i.p./Day) for 2 months. Taurine + ICE group in which rats were supplemented with (200 mg·kg−1, i.p. Taurine + ICE 15 mg/kg b.w./day orally) for 2 months. ResultThe findings indicated that Ipomoea persuaded an increase in lipid peroxidation markers as well as a disruption in antioxidant homeostasis, along with brain and serum cholinesterase (AChE) elevated levels, tumor necrosis factor (TNF), total creatine phosphokinase (CPK), creatine phosphokinase isoenzymes BB (CPK-BB), and lactate dehydrogenase (LDH). A considerable drop (P < 0.05) in the brain superoxide dismutase (SOD), catalase (CAT), and GSH in Ipomoea carnea. SOD, CAT, and GSH levels were enhanced in the Taurine co-treated group but did not achieve control levels. Ipomoea Furthermore, Induced apoptosis in brain tissue by Ipomoea carnea reflected in pro-apoptotic Bax, P53, and caspase 3 overexpression., while taurine upregulated the anti-apoptotic Bcl2 in addition to Histopathologic, electron microscope, and immunohistological brain examination finding support our finding concerning the protective role of taurine. ConclusionOur study proved the protective role of taurine as a supplement could recover Ipomoea carnea -induced oxidative changes and brain damage.

Ectopic thyroid tissue in the iris: a case report

BackgroundEctopic thyroid tissue in the iris, also known as a thyroid glandular epithelial choristoma of the iris, has only been described twice in the literature. In both cases it remained asymptomatic.Case presentationA 67-year-old female patient presented for the first time in mid-2017 with corneal endothelial decompensation, with a history of complicated cataract surgery and IStent® implantation. Slit lamp microscopy showed endothelial decompensation, pseudophakia, anterior synechiae and a whitish iris tumour adhering to the endothelium. The latter had existed since childhood. Given these findings, reduced visual acuity of hand movement perception and an intraocular pressure of 23 mmHg, we performed a keratoplasty combined with an en bloc resection of the iris tumour at 9 o’clock and sector iridectomy at the end of 2019. Histological and immunohistological examination of the iris tumour unexpectedly revealed thyroid tissue. After the procedure described above, the patient had an increase in visual acuity while the graft stayed clear and the eye showed no evidence of tumour recurrence or other complications.ConclusionsWe report a third case of ectopic thyroid tissue in the iris. Both previous cases remained asymptomatic, whereas in our case, size and location of the ectopic thyroid tissue contributed to a more complex cataract surgery resulting in endothelial decompensation. Therefore, in such cases appropriate patient information should be provided prior to cataract surgery. Furthermore, careful histological examination and examination of the thyroid is important to exclude malignant diagnoses such as a metastasis of a follicular thyroid carcinoma.

Intermittent intestinal intussusception in patients with mesenteric lymphadenitis in the ileocecal angle

Introduction. At present, lymphadenopathy – as a cause of idiopathic intestinal intussusception in little children - comes to the fore. However, intestinal intussusception in case of destructive mesenteric lymphadenitis in combination with lymphoid hyperplasia of the ileocecal angle ( what does not exclude lymphoma intraoperatively) is extremely rare in clinical practice.Purpose. To present a therapeutic and diagnostic algorithm in intestinal intussusception which was caused by the “volumetric“ abdominal lymphadenopathy.Case Report. A 3-year-old boy was admitted by the ambulance with recurrent abdominal pain, single episode of vomiting and no stool for 4 days. The ultrasound examination revealed intermittent ileocecal intussusception. The risk factor which had triggered the disease was an acute respiratory viral infection complicated with purulent mesenteric lymphadenitis and lymphoid hyperplasia with ulcerative lesions of the mucous membrane in the ileocecal junction. Laparoscopic desinvagination with lymph node removal and colonoscopy with ladder biopsy were performed. A differential diagnostics of nonspecific lymphadenopathy, Burkitt’s lymphoma and Crohn’s disease was made. Immunohistological examination intraoperatively was made; examination of the biopsy material excluded tumor and inflammatory bowel disease (IBD). The inflammatory process regressed under conservative therapy (antibacterial, anti-inflammatory and antispasmolytic) ; there were no any recurrent intussusception episodes within one-year follow-up.Conclusion. Successive ultrasound, laparoscopic and colonoscopic examinations followed by the immunohistological analysis of intraoperative and biopsy material allowed to put a correct diagnosis and, thereby, to minimize surgical aggression in ileocecal intussusception which was accompanied by volumetric lymphadenopathy.

Therapeutic Effect of IL-4 Receptor-Targeting Pro-Apoptotic Peptide (AP1-ELP-KLAK) in Glioblastoma Tumor Model.

BackgroundThermal-responsive self-assembled elastin-like polypeptide (ELP)-based nanoparticles are an emerging platform for controlled delivery of therapeutic peptides, proteins and small molecular drugs. The antitumor effect of bioengineered chimeric polypeptide AP1-ELP-KLAK containing an interleukin-4 receptor (IL-4R) targeting peptide and pro-apoptotic peptide (KLAKLAK) was evaluated in glioblastoma (GBM) in vitro and in vivo.Methods and ResultsHerein, the therapeutic effect of AP1-ELP-KLAK was tested in advanced, and less curable glioblastoma cells with higher expression of IL-4R. Glioblastoma cell lines stably expressing different reporter systems i.e., caspase-3 sensor (surrogate marker for cellular apoptosis) or effluc/enhanced firefly luciferase (cellular viability) were established to measure cell death non-invasively. Bioluminescence imaging (BLI) of D54/effluc and U97MG/effluc treated with AP1-ELP-KLAK exhibited higher cell death up to 2~3-fold than the control. Treatment with AP1-ELP-KLAK resulted in time-dependent increase of caspase-3 sensor BLI activity in D54/C cells and D54/C tumor-bearing mice. Intravenous injection of AP1-ELP-KLAK dramatically reduced tumor growth by inducing cellular apoptosis in D54/effluc tumor-bearing mice. Further, the immuno-histological examination of the excised tumor tissue confirmed the presence of apoptotic cells as well as caspase-3 activation.ConclusionCollectively, AP1-ELP-KLAK effectively induced cellular apoptosis of glioblastoma cells and non-invasive imaging provides a window for real-time monitoring of anti-tumor effect with the provision of improving therapeutic efficacy in a glioblastoma mice model.

Case report of portal hepatic schwannoma: presentation of multimodality images

BackgroundPortal hepatic schwannoma is a rare benign tumor and difficult to diagnose preoperatively because of its rarity and imaging manifestations that mimic malignancy. We present a case of portal hepatic schwannoma that showed moderate contrast enhancement on computed tomography (CT), extension along the bile duct on T2-weighted imaging and magnetic resonance cholangiopancreatography (MRCP), and uptake of 18F-fluorodeoxyglucose (FDG) on positron emission tomography.Case presentationUltrasonography at an annual health checkup identified a hepatic mass in a 38-year-old woman. CT showed a well-defined portal hepatic tumor with mild contrast enhancement. T2-weighted imaging and MRCP showed a clavate tumor extending along the intrahepatic bile ducts but no dilatation of the ducts. The tumor exhibited increased FDG uptake, such as maximum standardized uptake values of 5.0 and 6.5 in the early and late phases, respectively. Neither dilatation of intrahepatic bile ducts nor lymphadenopathy was identified, and the multimodality imaging suggested hepatic portal lymphoma, gastrointestinal tumor, or IgG4-related disease rather than cholangiocarcinoma. A needle biopsy via endoscopic ultrasonography was performed, and immunohistology confirmed the tumor as a schwannoma.ConclusionsThe diagnosis of a portal hepatic schwannoma requires immunohistological examinations in addition to multimodality imaging studies to reflect fully the pathohistological characteristics of the tumor.

Mesenchymal stem cells induce tumor stroma formation and epithelial‑mesenchymal transition through SPARC expression in colorectal cancer.

Tumor-stroma interactions serve a crucial role in the development of colorectal cancer (CRC), in which secreted protein acidic and rich in cysteine (SPARC) has been implicated. Due to interactions between cancer and stromal cells [mesenchymal stem cells (MSCs)], SPARC gene expression is markedly upregulated in CRC cells. The present study investigated the role of SPARC in CRC development and its potential as a biomarker. Specifically, the present study examined the association between SPARC expression and clinicopathological characteristics in 42 cases of CRC. SPARC expression in cancer cells was associated with T grade, N grade (TNM classification), stage and poor prognosis. Furthermore, the area of fibroblast-activating protein-positive staining around the cancer cells was increased in SPARC-positive compared with SPARC-negative cases. Proliferation and wound healing assays in SPARC-silenced KM12SM cells [short hairpin RNA SPARC (shSPARC)], the reduced SPARC expression of which was demonstrated by reverse transcription-quantitative PCR, revealed that the proliferative and migratory capacity of shSPARC cells did not differ from that of wild-type (WT) cells. However, it was markedly reduced when co-cultured with MSCs. Furthermore, in vivo, immunohistological analysis and RNA sequencing were conducted in an orthotopic implanted mouse model. Tumor growth and lymph node metastasis were markedly suppressed in shSPARC-transplanted tumors compared with WT-transplanted tumors, with a more marked suppression observed following shSPARC co-transplantation with MSCs. Immunohistological examination further revealed that the stromal reaction and epithelial-mesenchymal transition (EMT) were markedly suppressed in tumors co-transplanted with shSPARC and MSCs, and these results were consistent with RNA sequencing using RNA extracted from orthotopic tumors. Overall, these results suggested that SPARC expression in CRC cells is dependent on the interaction between cancer cells and stromal cells to induce EMT and promote stromal formation in the tumor microenvironment, suggesting its suitability as a novel target molecule for CRC treatment.

Quality assurance in dMMR and MSI diagnostics

Deficient mismatch repair (dMMR) and microsatellite instability (MSI) have therapeutic relevance not only for colorectal carcinomas but also for carcinomas of other entities (endometrium, biliary tract, pancreas). In order to guarantee the knowledge and good technical quality necessary for adequate implementation of the corresponding analyses in pathology institutes, the Pathology Quality Assurance Initiative ("Die Qualitätssicherung-Initiative Pathologie") has been offering proficiency tests (PT) for years. It has been shown for the dMMR PT that various antibody clones from different manufacturers provide comparable results in immunohistological examinations, except for slight variations. The difficulty lies in the staining protocol (intensity of staining) and the interpretation of the staining results. The molecular pathological MSI PT has shown apositive trend at ahigh-quality level over the last three years. Success rates increased from 89 (2018) to 97% (2019/2020). The choice of assay, whether commercial or in-house tests with the designated cutoffs for this purpose, has not been shown to have asignificant impact on the PTs in the selected EQA samples.

Immunohistological Examination of AKT Isoforms in the Brain: Cell-Type Specificity That May Underlie AKT's Role in Complex Brain Disorders and Neurological Disease.

Protein kinase B (PKB/AKT) is a central kinase involved in many neurobiological processes. AKT is expressed in the brain as three isoforms, AKT1, AKT2, and AKT3. Previous studies suggest isoform-specific roles in neural function, but very few studies have examined AKT isoform expression at the cellular level. In this study, we use a combination of histology, immunostaining, and genetics to characterize cell-type-specific expression of AKT isoforms in human and mouse brains. In mice, we find that AKT1 is the most broadly expressed isoform, with expression in excitatory neurons and the sole detectable AKT isoform in gamma-aminobutyric acid ergic interneurons and microglia. By contrast, we find that AKT2 is the sole isoform expressed in astroglia and is not detected in other neural cell types. We find that AKT3 is expressed in excitatory neurons with AKT1 but shows greater expression levels in dendritic compartments than AKT1. We extend our analysis to human brain tissues and find similar results. Using genetic deletion approaches, we also find that the cellular determinants restricting AKT isoform expression to specific cell types remain intact under Akt deficiency conditions. Because AKT signaling is linked to numerous neurological disorders, a greater understanding of cell-specific isoform expression could improve treatment strategies involving AKT.