Positive Immunohistochemical Reaction Research Articles

Hyperbaric Treatment Stimulates Chaperone-Mediated Macroautophagy and Autophagy in the Liver Cells of Healthy Female Rats.

The role of autophagy goes far beyond the elimination of damaged cellular components and the quality control of proteins. It also cleanses cells from inclusions, including pathogenic viruses, and provides energy-forming components. The liver, which is an organ with increased metabolism, is made up of cells that are particularly vulnerable to damage. Therefore, detoxification of liver cells in the process of autophagy has become a very important issue clinically. The aim of this study was an immunohistochemical evaluation of proteins activated in rat liver cells at different stages of hyperbaric autophagy. The rats used for the study were randomly divided into six equivalent groups-three control groups and three experimental groups. Animals from the experimental groups were subjected to hyperbaric treatment in a hyperbaric chamber, with a pressure of 1.6 ATA for 120 min. They breathed atmospheric air. Rats were decapitated within 5 or 10 days after removal from the chamber. Immunohistochemical reactions with beclin 1, LC3B, RAB7, and HSC73 proteins were carried out on preparations made from liver slices. A three-step labeled streptavidin-biotin detection method of paraffin blocks (LSAB three-step) was used for immunohistochemical research. The results were evaluated using computer programs for morphometric analysis of microscopic images by calculating the mean surface areas occupied by a positive immunohistochemical reaction in individual groups for all antibodies tested. Increased closure of substrates in the autophagosome (beclin 1) induced late endosome transport and accelerated autophagosome maturation process (RAB7). Furthermore, a larger number of autophagosomes (LC3B) was observed in liver cells immediately after the cessation of hyperbaric activity; however, this decreased after 5 days. During this time, chaperone-mediated autophagy (HSC73) was observed on a larger scale. This means that increased macroautophagy induced by hyperbaric treatment weakens with time that has elapsed since the cessation of high pressure, whereas similarly induced chaperone-mediated autophagy intensifies over time.

Immunohistochemical Identification and Assessment of the Location of Immunoproteasome Subunits LMP2 and LMP7 in Acquired Cholesteatoma.

Cholesteatoma, accompanied by chronic inflammatory response, is characterized by invasive growth and osteolytic activity. As specific proteasome isoforms, the immunoproteasomes serve as an important modulator of inflammatory responses. The aim of the present study was to determine the biological activity of cholesteatoma through the analysis of the expression and localization of immunoproteasome subunits of low molecule weight protein (LMP) 2 and LMP7. Cholesteatoma specimens were obtained from 15 adults who underwent ear surgery due to acquired attic cholesteatoma. Normal skin specimens were taken from retro-auricular skin incisions from the same patients. The specimens were stained with anti-LMP7 antibody, using immunohistochemistry techniques based on the binding of biotinylated secondary antibody with the enzyme-labeled streptavidin and the Envision FLEX system. In all specimens of cholesteatoma, the immunohistochemical reaction with the antibody against the LMP2 was positive, in both the cytoplasm of the cholesteatoma matrix and the perimatrix. A negative reaction with anti-LMP2 was observed in the cytoplasm and nuclei of control skin cells. A positive nuclear and cytoplasmic immunohistochemical reaction with anti-LMP7 has been demonstrated in numerous cells, in both the matrix and perimatrix of cholesteatoma. We present evidence of the presence of expressions of LMP2 and LMP7 within cholesteatoma tissue. Our results might bring new information concerning immunoproteasome-dependent pathophysiologic mechanisms in cholesteatoma.

Immunohistochemical Detection of the Expressed BRCA1 and BRCA2 Proteins in Microenvironment of Malignant Breast Cancerous Tissues Infected with Human Mammary Tumor Virus.

The objective of the study was to examine the prevalence of HMTV infection, its associations with breast malignant tissues, and the expression of BRCA1 and BRCA2 proteins. One hundred archival breast tissues, 40 biopsies from female patients with breast cancer (BC), and 20 healthy breast tissues from the control group were used in the study. Immunohistochemical analysis was conducted to detect the expressed BRCA1 and BRCA2 proteins. Digoxigenin-labeled HMTV probes were used in chromogenic in situ hybridization for the identification of HMTV in breast tumor tissues. The complementary sequence sites of the HMTV probe sequences were stained by NBT/BCIP as blue signals. There were 12 out of 40 (30%) benign breast tumorous tissues and 14 out of 40 (35%) BC tissues, while healthy control breast tissues were 10% (2 out of 20 tissues). Positive immunohistochemical (IHC) reactions for BRCA2 protein were observed in 12 out of 40 BC tissues (30.0%), 25% of benign breast tumorous tissues, and 5% of the control group. A significant (p < 0.05) statistical difference in the percentages of HMTV in the studied groups was found. HMTV might contribute to the development of subsets of benign and malignant breast tumors. The observed rates of defective or mutated BRCA1 and BRCA2 genes in healthy tissues indicate a role in the development of breast tumors.

Unusual Canine Cutaneous Melanoma Presenting Parietal Bone Metastasis: A Case Report

Melanocytic tumour anatomic location is considered an important prognostic indicator. The cutaneous forms are generally considered benign and may show various biological behaviours. This work reports a rare case of canine cutaneous melanoma showing parietal bone metastasis. Bone invasion in melanocytic tumours is often described in oral or visceral melanomas, but not in cutaneous forms. The patient (dog, male, mixed breed, 12 years) was initially presented for the surgical removal of a cutaneous tumour located on the skin of the carpal region of the right forelimb. Four months after, the patient returned with enlarged lymph nodes and acute respiratory failure. The patient was euthanized due to a decline in physical condition. The necropsy showed metastases in the affected forelimb, regional lymph node, splanchnic organs, parietal bone and meninges. Histopathological examination of tumour tissue samples revealed a mixture of pigmented and non-pigmented spindle and epithelioid melanocytes, while according to immunohistochemistry, the tumours showed a strong immunopositivity for VEGF and MMP-10, and a moderate positivity for MMP-2 expression. This case shows that cutaneous melanocytic tumours may show an aggressive malignant form with positive immunohistochemical reactions for multiple invasiveness factors.

6-Monoacetylmorphine-antibody distribution in tissues from heroin-related death cases: An experimental study to investigate the distributive response.

Heroin, a semisynthetic opioid drug synthesized from morphine, is the 3,6‐diacetyl ester of morphine (diacetylmorphine). The post‐mortem diagnosis of heroin‐related death could be an issue and usually rely on a combination of investigations, including the autopsy, histological and toxicological analysis. We conducted the present study to evaluate the correlation between the heroin concentration in biological fluids (peripheral blood, bile and urine) and the post‐mortem anti‐6‐MAM antibody expression in various tissues (brain, heart, lung, liver and kidney) using immunohistochemical staining. A quantitative analysis of the immunohistochemical reaction was carried out. 45 cases of heroin‐related death investigated at the Forensic Pathology Institutes of the University of Rome, Foggia and Pisa were included. The control group was composed of 15 cases of death due to other causes, without brain lesions and negative toxicological analysis for drugs. We found a positive immunohistochemical reaction in different organs and it was related to the timing of heroin metabolization. No reaction was found in the control group. Our findings show that immunohistochemistry can be a valuable tool for the post‐mortem diagnosis of acute heroin abuse. A better understanding of the timing of heroin's metabolism can be useful in the forensic field and for future therapeutic applications.

Chosen Antioxidant Enzymes GPx4 and GPx8 in Human Colorectal Carcinoma: Study of the Slovak Population.

Background and Objectives: Nowadays colorectal carcinoma (CRC) is one of the most common causes of death in patients with malignant neoplasms worldwide. Our work aimed to determine the possible involvement of glutathione peroxidases 4 and 8 (GPx4 and GPx8) in this specific tumor process. Materials and Methods: The expression of GPx4 and GPx8 in 58 specimens of human colorectal cancer tissues and normal tissues was detected by the indirect immunohistochemical method under a light microscope. Statistical analysis was done by Chi-squared test. Histological findings were compared with data such as gender, age, tumor grade, histotype and lymph nodes alteration. Results: In all specimens of healthy tissue the presence of both, GPx4 and GPx8, was detected in the cytoplasm of epithelial cells. On the other hand, a positive immunohistochemical reaction against GPx4 only in 41.4% and against GPx8 only in 29.3% of human colorectal adenocarcinoma specimens were observed. Any significant difference between the presence of GPx and the age, the gender of the patient, tumor grade, histotype of cancer and the lesion of regional lymph nodes has not been detected. Conclusions: Our foundation could mean, that GPx4 and GPx8 have no important role in CRC pathogenesis, but the loss of these enzymes probably indicates a serious pathological process ongoing in the large intestine. To our knowledge, this is the first paper describing GPx8 presence in human colorectal carcinoma.

The effect of biodegradable silk fibroin-based scaffolds containing glial cell line-derived neurotrophic factor (GDNF) on the corneal regeneration process

Corneal injury due to ocular trauma or infection is one of the most challenging vision impairing pathologies. The aim of the work was to study the effect of biodegradable silk fibroin-based scaffolds containing GDNF on the corneal regeneration process. During cultivate the highest keratocytes proliferative activity was registered with scaffolds containing 250 ng/ml and 500 ng/ml GDNF. In mice with an experimental model of epithelial-stromal damage to the cornea, silk fibroin-based scaffolds containing GDNF in various concentrations were used (in groups 1, 2 and 3 silk fibroin-based scaffolds containing GDNF in a concentration of 50 ng/ml, 250 ng/ml and 500 ng/ml, respectively; in group 4 - silk fibroin-based scaffolds without GDNF; in group 5 - a solution of GDNF with concentration of 500 ng/ml; group 6- control). The area of the corneal epithelial defect in groups 2, 3, and 5 was less than in the other groups. The most pronounced positive immunohistochemical reaction with antibodies to Bcl2, Bax, phosphoERK1/2 and phospho-JNK1/2, Ki67, Gap43 was observed in groups 2 and 3. Thus, silk fibroin-based scaffolds with GDNF stimulate the epithelialization process, proliferative activity of epithelial cells and keratocytes, accelerate the formation of the stromal nerve plexus and exhibit anti-apoptotic activity.

Determination of postmortem interval through histopathological alterations and collagen evaluation in the prostate of Wistar albino rats

Background: Determination of postmortem interval (PMI) has always been a fundamental issue in forensic fields. Therefore, the purpose of this study was to assess usefulness of histopathological changes and collagen degradation in the prostate for estimation of PMI in adult male rats. Methods: Forty male albino rats were euthanized using CO2 chamber and then classified randomly into 8 groups; each group consisted of 5 rats. Rats were used for determining postmortem changes at 0, 6, 12, 24, 36, 48, 60 and 72 hs. Sections of prostate were fixed and stained for histopathology and quantitative analysis of collagen by Masson trichrome. Moreover, slices of 5µm mounted on positively charged slides were carried out for collagen III immunohistochemistry. Results: The prostate showed normal histology at 0 and 6 hs postmortem (PM). While, at 12 hs, most sections of the gland revealed no abnormalities except for mild focal widening between the acini. Starting from 24 hs, epithelial desquamation was seen in most acini. Seventy-two hours, obvious necrosis of prostatic acini appeared in most of samples. Degradation of collagen started and the spaces between acini became wide at 12 and 24 hs PM. Complete loss of stain reactivity was observed at both 60 and 72 hs. Moreover, a significant decrease in the amount of collagen’s stained areas in the prostate started from 12 hs. Positive immunohistochemical reaction of collagen III was detected at 0, 6 and 12 hs PM. However, it was completely lost at 60 and 72 hs. Interestingly, the immuno-reactive area of collagen III was significantly increased at 6 hs, then a remarkable decline in immuno-reactive area was observed at 12 hs. Conclusion: Collagen type III proved to be a successful parameter with histopathology in determination of PM interval. However, further studies still needed to confirm the accuracy of these parameters.

The Possible Role of Orthoboric Acid on Adipogenesis in Adult Male Albino Rat. Histological and Immunohistochemical Study

Background: obesity is a worldwide problem which leads to increased morbidity and rate of deaths. Clinical use of some therapeutic options for the prevention of overweightness is limited due to their dangerous side effects. Orthoboric acid is a new safe and beneficial treatment for prevention of high fat diet induced obesity. Aim: study the effect of orthoboric acid on adipogenesis in rats. Materials and methods: the study included 30 adult male albino rats. The rats were divided into 4 main groups. Group I: control. Group II (High Fat Diet ′′HFD′′ group): IIa (HFD 8 weeks), IIb (HFD 12 weeks). Group III (HFD and orthoboric acid group) for 8 weeks. Group IV (HFD for 12 weeks but starting from the 8th week the rats were given orthoboric acid daily for 4 weeks). Orthoboric acid was used at dose of 2.5 mg given orally via intragastric tube. Body weights of all rats were recorded weekly during the experiment. Adipose tissue specimens were collected at the end of experiment and processed for H&E, oil red O stain and anti β - catenin immunohistochemistry. Morphometric and statistical analysis were also done. Results: HFD group showed significant increase in body weight and in size of adipocytes compared to the control group meanwhile, groups of orthoboric acid showed a significant decrease in body weight and size of adipocytes relative to HFD group. As regard effect of orthoboric acid on adipogenesis, groups treated with orthoboric acid showed positive immunohistochemical reaction for β- catenin with subsequent decrease in lipid accumulation confirmed with less optically dense oil red O staining compared to HFD group. Conclusions: low dose of oral orthoboric acid was able to reduce body weight. Therefore, it can be considered as a suitable treatment for obesity due to its vital role in inhibition of adipogenesis.

DLL3 (delta-like protein 3) expression correlates with stromal desmoplasia and lymph node metastases in medullary thyroid carcinomas.

Medullary thyroid carcinomas (MTC) are rare and aggressive neuroendocrine tumors of the thyroid. About 70% of MTC are sporadic; approximately 50% of those harbor somatic RET mutation. DLL3 is widely expressed in many neuroendocrine tumors and has been evaluated as a potential therapeutic target. Since stromal desmoplasia in sporadic MTC has been identified as a reliable predictor of aggressive behavior and development of lymph node metastases, a possible correlation of DLL3 expression with the presence of stromal desmoplasia was of particular interest. 59 paraffin-embedded samples of sporadic MTC with (44 cases) and without (15 cases) stromal desmoplasia and known lymph node status were included. DLL3 expression was determined by immunohistochemistry; no expression (0%), low expression (1–49%) and high expression (≥50%) were correlated with clinicopathological data. The proportion of DLL3 positivity was significantly correlated with both stromal desmoplasia (P < 0.0001) and lymph node metastases (P < 0.0001). MTC without stromal desmoplasia consistently lack DLL3 expression. This is the first study to focus on MTC regarding DLL3 expression and the relationship to various factors. Our results demonstrate that expression of DLL3 in MTC represents a reliable surrogate marker for stromal desmoplasia and lymph node metastases and might be an indicator for aggressive clinical behavior. DLL3 expression in ≥50% of tumor cells virtually excludes MTC without stromal desmoplasia. DLL3 was discussed as a potential therapeutic target in malignant tumors of other locations with positive immunohistochemical reaction and might therefore be a new therapeutic option in MTC, as well.

EXPRESSION FEATURES OF HUMAN PAPILLOMA VIRUS TYPE 16 AND ANTI-EPSTEIN-BARR VIRUS IN PLEOMORPHIC ADENOMA, SURROUNDING AND INTACT SALIVARY GLAND

The aim is to reveal the expression features of MCA to human papilloma virus type 16 and anti-Epstein-Barr virus in the pleomorphic adenoma, surrounding and intact salivary gland. Materials and methods: It was used surgical and biopsy material from 30 patients, represented by pleomorphic adenomas with surrounding to tumor tissue of the salivary gland and intact tissue of the salivary gland (the distance between the tumor and the intact salivary gland – 10 mm). Immunohistochemical study was performed using mouse monoclonal antibody (MCA) to human papilloma virus type 16 (clone CAMVIR-1, «Diagnostic BioSystems», USA) and anti-Epstein-Barr virus (LMP, clone CS. 1-4, «Dako», Denmark). Visualization was performed, using an EnVisionTM FLEX detection system (Dako, Denmark). Antigen unmasking was carried out in citrate buffer pH 6.0 at 95°C. Primary antibodies were incubated at room temperature for 30 minutes, secondary antibodies – 20 minutes. Sections were counterstained with Gill hematoxylin. We assessed the immunohistochemical reaction by a semi-quantitative method by counting the percentage of positively stained cells in the field of view of a microscope × 400. Microspecimens were studied and photoarchived on an Olympus BX-41 microscope (Japan). Results: In this study it was detected a positive immunohistochemical reaction with MCA to human papilloma virus type 16 and anti-Epstein-Barr virus, respectively, in 26 (86.7%) and 8 (26.7%) cases. Epithelial, mixed and mesenchymal variants of pleomorphic adenoma of the salivary glands are characterized, respectively, by the severely expressed, moderately expressed and minimally expressed of MCA to human papilloma virus type 16 and anti-Epstein-Barr virus. The parenchymal component of pleomorphic adenoma is characterized by more marked expression of these markers as compared to the stromal component. The epithelial cells of the salivary glands, surrounding the pleomorphic adenoma, as well as intact salivary glands, express MCA to human papilloma virus type 16 and anti-Epstein-Barr virus. The severity of the expression of these markers in the salivary gland is determined by the histological variant of the tumor (severely expressed in the epithelial variant, moderately expressed in the mixed variant, and minimally expressed in the mesenchymal variant). Conclusions: The immunohistochemical study has shown that the Epstein-Barr virus and, especially, human papilloma virus type 16 can act as exogenous trigger factors involved in the development of pleomorphic adenoma of the salivary glands. The revealed immunohistochemical features of MCA expression to human papilloma virus type 16 and anti-Epstein-Barr virus in the salivary gland surrounding the pleomorphic adenoma and in the intact tissue of the salivary gland make it possible to recommend the extracapsulardissection of the tumor with resection of the adjacent intact tissue of the salivary gland at a distance of 10 mm in patients with pleomorphic adenoma.

Immunohistochemical Diagnosis Method in Parvovirus

10 cases of histopathologically suggestive lesions for parvovirus in the canine species were examined by the avidin-biotin-peroxidase complex. For this, a mouse monoclonal antibody was used. Following the immunohistochemical examination, positive results were obtained in 80% (8 dogs) of processed samples. The most frequent positive immunohistochemical reaction was observed in macrophages, lymphocytes, necrotic cells of the intestinal crypt, evidence of parvoviral antigen is reliable for determining the etiology of the disease if parvovirus is suspected on the basis of the lesions. In order to perform the diagnostic immunohistochemical procedure, the strepto-avidin-biotin-peroxidase method was used, using the monoclonal antibody. It is of murine origin. Negative cases of parvovirus by immunohistochemical examination show the existence of other causes that produce the same lesions; coronaviruses, rotaviruses or coronaviruses associated with clostridium perfringens which may cause intestinal necrosis of the crypt. All dog breeds can do this disease, but in the Poodle breed the disease has been more common. Sex is not a criterion of receptivity, but in proportion of over 60% were affected females. Regarding the seasonal evolution, the disease is more common in the winter period compared to the other seasons.

Changes in the Expression of TLR2 During the Intestinal Phase of Trichinellosis.

IntroductionToll-like receptors (TLRs) play an important role in fast activation of the immune response to a variety of pathogens, including parasites. In this study, we focused on TLR2, because this receptor is one of the best known and most frequently analysed members of the TLR family. The aim of this study was to assess the effect of Trichinella spiralis on expression of TLR2 during the intestinal stage of infection.Material and MethodsThe experimental material consisted of isolates prepared from the intestines (jejunum and colon) of BALB/c mice infected with T. spiralis taken at 4, 8, and 16 days post infection.ResultsOur results based on quantitative real-time PCR showed that the mRNA level for TLR2 was statistically significantly higher in the jejuna of mice infected with T. spiralis than in this tissue of uninfected mice. In addition, the presence of TLR2 protein in the intestinal phase of trichinellosis was confirmed by a strong positive immunohistochemical reaction.ConclusionOur results indicate that infection with T. spiralis changes the expression of TLR2 in the small intestine of the mouse host and suggest a contribution of these receptors to the host defence mechanisms during experimental trichinellosis.

Malignant Peripheral Nerve Sheath Tumor and Hypertrophic Osteopathy in a Bitch

Background: Malignant peripheral nerve sheath tumors (MPNSTs) are rare, aggressive, relapsing, metastatic cutaneous neoplasms of mesenchymal origin. So far, no account on the association of this disease with hypertrophic osteopathy (HO) in dogs is available in the literature. Current theories on the possible causes of HO suggest that this disease may be triggered by a primary neoplasm as well as by its metastasis. The objective of this work is to report the clinical, cytological, radiographic, histopathological, and immunohistochemical aspects of a dog affected by MPNST and HO.Case: A 13-year-old female mongrel dog was presented with a history of ulcerated nodular lesion on the skin of the flank. Several ulcerated and non-ulcerated tumors were observed on the thorax, neck, and head at the physical exam. Cytological examination of the nodules revealed presence of mesenchymal cells with a malignant aspect. Complete blood count revealed anemia. There were no alterations in the biochemical tests performed. Thoracic radiographs showed presence of a nodular interstitial pattern in the cranial, medial, and caudal lobes of the lungs. To improve quality of life of the patient, the veterinary team opted for surgical excision of the tumoral ulcerations. Slight claudication in the pelvic limbs was noticed prior to the surgery. Histopathological analyses of the excised nodules verified the existence of malignant mesenchymal neoplasia, which was categorized as peripheral nerve sheath tumor after immunohistochemical examination. Additional cutaneous tumors emerged after surgical excision, along with increased claudication, edema, and pain in the legs. Complete blood count revealed persistent anemia, lymphopenia, monocytosis, and neutrophilia. Radiographs showed an increase in the number and size of the nodules. Radiography of the limbs showed presence of palisading periosteal reaction, and increased radiopacity and volume in the adjacent soft tissues. Doxorubicin and vincristine were administered, but the patient died one week after commencement of the treatment.Discussion: Cytology suggested presence of a malignant neoplasm of mesenchymal origin, and the histopathological and immunohistochemical exams substantiated the diagnosis of malignant peripheral nerve sheath tumor. Macroscopic and histopathological examinations revealed characteristics similar to those described for MPNST in the literature. Positive immunohistochemical reactions for the markers NSE and CD56 determined the diagnosis of MPNST in spite of being negative for S-100, which could represent a consequence of variability in expression and cell differentiation. MPNST exhibits a variable distribution among breeds; it is more predominant in male and elder dogs. Animals exhibiting neoplastic lesions in the thorax, abdomen, head and neck show a high incidence of metastasis after surgical removal of these lesions. The clinical signs of MPNST were unspecific; the alterations observed in the four limbs were observed concomitantly with the diagnosis of hypertrophic osteopathy (HO), and could be due to this condition rather than MPNSC. Histopathological analysis of the pulmonary neoplasms was not possible. However, there is strong suspicion that the pulmonary lesions were a consequence of MPNSC metastasis, which could be the underlying cause of the reemergence of the cutaneous nodules. We believe that HO was caused by the thoracic tumors because the clinical signs became more intense after the increase in the nodules found in the lungs. The difficult diagnosis of MPNSC could be the reason for this type of tumor is rarely identified. MPNSC should always be included in the differential diagnosis for cutaneous tumors, and the possibility that HO represents a consequence of these tumors should be considered.

Peritumoral Clefting and Expression of MMP-2 and MMP-9 in Basal Cell Carcinoma of the Skin.

Peritumoral clefting is one of the main histologic features of basal cell carcinoma of the skin (BCC). The aim of the study was to analyze the expression of MMP-2 and MMP-9 both in cells of basal cell carcinoma and in the adjacent stroma and to correlate the findings of immunohistochemical analysis with the presence of peritumoral clefting. The study was made on archival material comprising 48 cases of BCC. These were scanned for the presence of peritumoral clefts. The results of immunohistochemical staining for MMP-2 and MMP-9 were determined semiquantitatively using immunohistochemical staining index (ISI). Peritumoral retractions were found in 40 BCC cases. Positive immunohistochemical reaction for MMP-2 in tumor cells was found in 47 cases and in all cases in the adjacent stroma. Positive immunostaining for MMP-9 in BCC tumor cells was observed in 37 cases and in all cases in the adjacent stroma. There was no statistically significant association between peritumoral retractions and expression of MMPs. A statistically significant correlation was found in the expression of both MMP-2 and MMP-9 between the tumor and the stroma. Tumor cells elaborate MMP-2 and -9, but they also produce some other factors that may induce production of MMPs in adjacent stromal cells. The role of MMPs in the development of peritumoral clefts could not be confirmed.

Strategy for prevention of uterine leiomyoma recurrence in combination with adenomiosis after organ-conserving operations in reproductive aged women

Aim of the study: to reduce the number of relapses and improve the reproductive function of women after surgical removal of uterine leiomyoma (LM).Materials and methods: the study included 76 women of reproductive age who underwent organ-preserving surgical treatment for LM. Women were divided into two groups depending on the adenomyosis presence. Group I included 35 women with isolated LM, group II included 41 women with LM in combination with adenomyosis. Groups I and II were divided into 4 subgroups: in Ia and IIa subgroups during preparation for surgery patients received one of the agonists of gonadotropin releasing hormone (GnRH) for 3 months. In Ib and IIb subgroups suppressive therapy was not performed prior to surgical treatment. After a myomectomy women of I and II groups received one of the GnRH agonists for the first three months, followed by the low-dose combined oral contraceptive. The choice of funds was justified on the results of an immunohistochemical study of the Ki-67 antigen expression (proliferation marker), estrogen and progesterone receptors, and cyclooxygenase-2 (COX-2). Group IIb women were additionally prescribed non-hormonal therapy with fibrinolytic (streptokinase) and proteolytic (streptodornase) enzymes, as well as herbal antiproliferative agent containing indole-3-carbinol and epigallocatechin-3-gallate.Results of the study: an immunohistochemical study showed that in women who did not receive suppressive therapy before surgery (Ib and IIb groups) Kі-67 antigen expression in the tumor nodes was significantly increased. Almost 2 times greater was the difference between the Ki-67 antigen expressions in the LM nodes with the adenomyosis combination. GnRH agonists did not inhibit the expression of estrogen and progesterone receptors. COX-2 expression was significantly higher than in intact myometrium. In women with combined uterine pathology the number of cells with a positive immunohistochemical reaction to COX-2 was 6.4 times higher than with isolated LM. Conclusions: myomectomy surgery only partially solves the problems of reproductive health restoration in women with combined uterine pathology. The proposed long-term suppressive therapy made it possible to reduce the tumor relapses number by almost 2.6 times after surgical organ-preserving treatment and to improve reproductive function in almost every third woman.

Intercellular adhesion in testicular germ cell tumors

Testicular germ cell tumors (TGCT) is one of the main causes of oncological mortality in young men. One of the factors determining the prognosis of cancer patients is the determination of molecular biological markers which correlate with a poor prognosis. The purpose of the work. To determine the state of intercellular adhesion in testicular postpubertal-type yolk sac tumor (YSTPT), postpubertal-type teratoma (TPT), spermatocytic tumor (ST) and teratoma with somatic-type malignancy (TSTM). Material and methods. The study was performed on the material of TGCT of patients who have been examined and treated at the Kharkiv Regional Clinical Center of Urology and Nephrology named after V.I Shapoval for the period covering 1998-2017. Groups of investigation were formed according to the pTNM classification of WHO. For assessment of the state of intercellular adhesion in TGCT the relative area (S) and intensity (L) of E-cadherin and β-catenin expression were studied. Results. During the tumorous progression S of E-cadherin expression in YSTPT and TPT was gradually decreasing right up to the complete reduction of this marker expression. These changes were combined with increasing of β-catenin expression. In the case of positive immunohistochemical reaction with E-cadherin and β-catenin in all investigated TGCT instead of the «typical» membrane localization there were «heterogeneities» which were also considered as a loss of intercellular adhesion. In ST the S and L of E-cadherin expression were higher than those in the corresponding groups of investigated TGCT but marker localization was also atypical. As for β-catenin, S and L of its expression were at the level of mean values in YSTPT and higher than analogous indexes in the corresponding group of TPT. Localization of β-catenin was atypical. In TSTM of both groups the reaction with E-cadherin was either negative or marker had an atypical localization. The same situation was observed at investigation of β-catenin. Conclusion. In YSTPT and TPT at tumorous progression a loss of intercellular adhesion occurs what leads to the dissociation of tumor cells, development of vascular invasion and metastatic disease.

Wounds as the Portal of Entrance for Parrot Bornavirus 4 (PaBV-4) and Retrograde Axonal Transport in Experimentally Infected Cockatiels (Nymphicus hollandicus).

In this study, we investigated the natural route of infection of psittacine bornavirus (PaBV), which is the causative agent of proventricular dilatation disease (PDD) in psittacines. We inoculated two infection groups through wounds with a PaBV-4 isolate. In nine cockatiels (Nymphicus hollandicus) we applied a virus suspension with a titer of 103 50% tissue culture infection dose (TCID50) via palatal lesions (Group P, P1-9). In a second group of three cockatiels, we applied a virus suspension with a titer of 104 TCID50 to footpad lesions (Group F, F1-3). In two cockatiels, the control (or "mock") group, we applied a virus-free cell suspension (Group M, M1-2) via palatal lesions. The observation period was 6 mo (Groups P and M) or 7 mo (Group F). We monitored PaBV-4 RNA shedding and seroconversion. At the end of the study, we examined the birds for the presence of inflammatory lesions, PaBV-4 RNA, and antigen in tissues, as well as virus reisolation of brain and crop material. We did not observe any clinical signs typical of PDD during this study. We also did not see seroconversion or PaBV RNA shedding in any bird during the entire investigation period, and virus reisolation was not successful. We only found PaBV-4 RNA in sciatic nerves, footpad tissue, skin, and in one sample from the intestine of Group F. In this group, the histopathology revealed mononuclear infiltrations mainly in skin and footpad tissue; immunohistochemistry showed positive reactions in spinal ganglia and in the spinal cord, and slightly in skin, footpad tissues, and sciatic nerves. In Groups P and M we found no viral antigen or specific inflammations. In summary, only the virus application on the footpad lesion led to detectable PaBV RNA, mononuclear infiltrations, and positive immunohistochemical reactions in tissues of the experimental birds. This could suggest that PaBV spreads via nervous tissue, with skin wounds as the primary entry route.

Three-dimensional evaluation of mucoepidermoid carcinoma on the hard palate

Mucoepidermoid carcinoma is of unknown aetiopathology, but it may be related to genetic factors, exposure to ionizing radiation and smoking habits as well. Mucoepidermoid carcinoma is considered the most common malignant neoplasm of salivary glands found in the oral cavity, with its clinical characteristics (e.g., size and shape) favoring the diagnosis and treatment planning. This report describes a case of mucoepidermoid carcinoma which infiltrates into the maxillary sinus and nasal fossa through the palate and highlights the importance of obtaining three-dimensional (3D) images of the tumor for volumetric analysis, thus improving the chances of a successful surgery. We have described the use of the segmentation technique in which the tumor’s volume was calculated to help in predicting surgical outcomes. A 50-year-old patient sought dental treatment because of a purplish swelling in the hard palate region on the left side. Computed tomography examination was performed for visualization of the lesion and incisional biopsy for obtaining a sample, which was sent to anatomopathological study. Histological characteristics of the lesion and presence of neoplastic cells showed positive immunohistochemical reactions for cytokeratin 7 antibody, leading to a diagnosis of mucoepidermoid carcinoma. In view of the malignant characteristics of the lesion, the patient was referred to a head and neck surgeon for treatment.

Case Report Plasmablastic Lymphoma

Plasmablastic lymphoma (PBL) is a rare and aggressive subtype of non-Hodgkin lymphoma that commonly occurs in human immunodeficiency virus (HIV)–positive individuals and affects oral sites. The aim of this work was to report a case of a 57-year-old female patient diagnosed with HIV infection, presenting with a purplish exophytic lesion, which was bleeding, painful, friable, located in the maxilla, and affecting the alveolar ridge of teeth #23 to #27, hard palate, and vestibule. Panoramic radiography was performed. Initially, Kaposi sarcoma was suspected. However, after incisional biopsy and positive immunohistochemical reactions for Ki-67 (high expression), LCA, plasma cell, CD138, and EBER and negative reaction for CD20 and CD34, the diagnosis of PBL was established. The patient's health status deteriorated rapidly, and shortly after the receiving the results of the analyses, she died before initiation of cancer treatment.