Imunohistohemijska identifikacija citokeratina u submandibularnoj pljuvačnoj žlezdi pacova u toku ontogeneze

Cytokeratins (CK) are a group of proteins comprised of at least 20 different skeletal polypeptides specific for simple and complex epithelia of almost all tissues. Until recently only a few antibodies for cytoskeletal proteins were available and thus it was very difficult to visualize all the components of postnatal cytodifferentiation of salivary glands. Therefore, morphologic and immunophenotypic features of the CK polypeptides, have been analized within the rat submandibular salivary glands (SSG) during postnatal development. SSG were obtained from 1, 30 and 60 days old, male, AO rats. Streptavidin-biotin immunoperoxidase method was used for immunohistochemistry. Cryostat sections were fixed in acetone and incubated with monoclonal antibodies (mAbs) which specifically react with CK polypeptides (CK7, CK8, CK18, CK kidney, K8.12, K8.13) and rat epithelium (PT13D11). Immunohistochemical analysis of the rat SSG showed phenotypic heterogeneity of particular components of this gland during postnatal development. Regarding CK profiles it was shown that epithelial cells of the acini expressed CK18, whereas epithelial cells of the duct expressed K8.12 and K8.13 mAbs. Epithelial cells of the acini were not stained by K8.12 and PT13D11 mAbs. Intercalated ducts were strongly CK kidney+ and PT13D11+. Myoepithelial cells of the acini were stained by K8.13 mAbs, whereas these cells were not stained by mAbs specific for simple epithelia. This immunohistological study showed that the heterogeneity expression of CK polypeptides might also be useful to further understand the origin of epithelial SSG cells, as well as their development and function.Citokeratini (CK) su grupa složenih proteina koji se sastoje od najmanje 20 različitih polipeptida citoskeleta, specifični za običan i složeni epitel skoro svih tkiva. Do skoro je bilo dostupno samo nekoliko antitela specifičnih za proteine citoskeleta i zbog toga je bila otežana vizualizacija komponenti postnatalne citodiferencijacije pljuvačnih žlezda. U ovoj studiji su, pomoću panela monoklonskih antitela (mAt), analizirane morfološke i imunofenotipske osobine CK polipeptida, u submandibularnoj pljuvač noj žlezdi (SPŽ) pacova tokom ontogeneze. Submandiblarne pljuvačne žlezde uzimane su od soja AO pacova, muškog pola, starosti 1, 30 i 60 dana. U ovom radu korišćena je imunohistohemijska metoda streptavidin-biotin peroksidaznog bojenja. Tkivni preseci fiksirani su u acetonu i inkubirani sa mAt specifičnim za CK polipeptide (CK7, CK 8, CK 18, CK bubrega, K8.12, K8.13) i epitel pacova (PT13D11). U ovoj imunohistohemijskoj analizi, ekspresija anti -CK antitela na SPŽ pacova, pokazuje fenotipsku heterogenost određenih komponenti ove žlezde tokom postnatalnog perioda. Epitelne ćelije acinusa su CK18+, dok epitelne ćelije kanalića imaju izražene CK definisane sa K8.12 i K8.13 mAt. Epitelne ćelije acinusa nisu bile obojene sa K8.12 i PT13D11 mAt. Umetnuti kanali su bili izrazito CK bubrega+ i PT13D11+. Mioepitelne ćelije acinusa su bile obojene sa K8.13 i PT13D11 mAt, dok su ove ćelije imale negativnu reakciju sa mAt specifičnim za obični epitel. Ova imunohistohemijska studija pokazuje heterogenost u ekspresiji CK polipeptida i može poslužiti za razumevanje porekla, razvoja i funkcije epitelnih ćelija SPŽ pacova

Intravascular large cell lymphoma: Clinicopathological and immunohistochemical description of a case

A 51-year-old female patient was admitted to the Neurological clinic because of motor seizures with myoclonus of the right hand and right side of the face. The results of initial brain CT scan, chest X rays, EEG ultrasonography of the great blood vessels and laboratory tests made in another hospital were unremarkable. Because of repeated partial seizures transient aphasic disturbances, urinary sphincter disturbances and periodic low-grade fever the patient was transferred to our hospital four months after the disease onset. Laboratory tests and NMR suggested a nonspecific disseminated viral encephalitis. After administration of Endoxan she was ambulatory for several weeks and then became increasingly exhausted confused, febrile, dyspneic, tachypneic and developed a shock status with hepatorenal insufficiency. She died after 7 months of disease duration. Postmortem examination revealed intravascular collections of large atypical lymphoid cells of B cell line. Blood vessels changed in this way were common in the brain and rare in other organs including skin, lungs, heart, liver spleen and digestive system. They were not found in the lymph nodes and bone marrow. A biopsy was not done because of absence of symptomatic and swollen tissues. However, correlation of clinical feature and postmortem findings shows that absence of clinical manifestations in an organ does not mean lack of microscopic pathological changes and biopsy should be done regardless of absence of clinical signs. This case shows that intravascular lymphoma may mimic vasculitis or disseminated nonspecific viral encephalitis

Traumatski neurom mentalnog nerva nakon ekscizije mukokele donje usne

Traumatic neuroma represents reactive hyperplasia of irregularly positioned Schwann neurofibril cells and connective tissue - stroma that develop as a result of unsuccessful attempts to regenerate peripheral nerve after injury. This paper presents the case of a patient with painless traumatic neuroma of the lower lip formed in the same place where he previously had surgically removed salivary cyst of minor salivary gland - mucocele. Histopathological examination confirmed the diagnosis of traumatic neuroma.Traumatski neurom predstavlja reaktivnu hiperplaziju iregularno postavljenih Švanovih ćelija i neurofibrila u vezivno-tkivnoj stromi koja nastaje kao posledica neuspešnih pokušaja regeneracije nakon povrede perifernog nerva. U ovom radu je prikazan slučaj pacijenta sa bezbolnim traumatskim neuromom donje usne koji se pojavio na istom mestu gde je prethodno hirurški uklonjena salivarna cista male pljuvačne žlezde donje usne - mukokela. Histopatološki nalaz je pokazao da je uklonjena promena bila traumatski neurom

Expression of matrix metalloproteinases and endogenous inhibitors in abdominal aortic aneurysm and aortoiliac occlusive disease (syndrome leriche)

© 2017 Charles University. All rights reserved. Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) play a complex role in the pathogenesis of atherosclerosis. We compared (1) the histopathological findings in patients with abdominal aortic aneurysms (AAA) and aortoiliac occlusive disease (AOD); (2) the expression of MMP-2/MMP-9 and TIMP-1/TIMP-2 in aortic layers, inflammatory cells and smooth muscle cells (SMCs), aiming to identify the common underlying pathogenic mechanisms of the disease development. Samples were obtained from 30 patients with AAA and 30 with AOD. Aortic histology and immunohistochemistry were performed to evaluate inflammatory changes and MMP and TIMP expression. Thrombosis and ulceration were more frequent in AOD than in AAA. The MMP-9 expression was elevated in all aortic layers of AAA patients and in media/adventitia of AOD patients, mainly followed by lower expression of its inhibitor TIMP-1. Higher MMP-9 expression was also found in SMCs and macrophages of both AAA and AOD specimens, while higher TIMP-1/TIMP-2 were predominantly observed in the lymphocytes and macrophages of the aneurysm. These results showed that both conditions exhibited increased MMP-9 expression; however, the MMP expression pattern differed to some degree between the aneurysms and occlusive disease. The variations in molecular mechanisms underlying dilatative/stenosing disease warrant further investigation

Immunohistochemical Analysis and Prognostic Value of Markers of the Intrinsic Pathway of Apoptosis in Adenoid Cystic Carcinoma of the Salivary Glands

Adenoidni cistični karcinom (ACC), je jedan od najčešćih malignih tumora pljuvačnih žlezda. Učestvuje sa oko 10-15% kod salivarnih neoplazmi. Karakteriše ga spor rast, visoka incidenca ka invaziji nerva, retko regionalno metastaziranje, česti lokalni recidivi i spor razvoj progresivnih i relativno indolentnih udaljenih metastaza. Najznačajniji faktori za prognozu ovog tumora su klinički stadijum i histološki izgled tumora, prisustvo ili odsustvo tumorskog tkiva na linijama resekcije, invazija nerva. Apoptoza je regulatorni mehanizam tkivne homeostaze. Poremećaj regulacije apoptoze je relativno česta karakteristika maligne ćelije. U zavisnosti od stimulansa, putevi aktivacije apoptoze mogu biti: receptorski (spoljašnji) i mitohondrijski (unutrašnji) put. U unutrašnjem putu apoptoze značajnu ulogu igraju Apaf-1, inicijatorne i efektorne kaspaze. Apaf-1 je multidomenski protein, za koji se vezuje citohrom c i prokaspaza 9 što dovodi do formiranja apoptozoma. Ovaj kompleks proteolitički aktivira inicijatornu kaspazu-9. U završnoj fazi apoptoze, aktivirana kaspaza 9 dovodi do aktivacije efektorne kaspaze 3, čijim ulaskom u jedro započinje degradaciona faza apoptoze. Cilj U ovoj studiji cilj je bio ispitati imunohistohemijsku ekspresiju Apaf-1, kaspaze 9 i 3 u ACC pljuvačnih žlezda, i rezultate korelirati sa kliničko-patološkim parametrima kako bi se utvrdilo koji je od njih nezavistan prognostički parametar. Materijal i metode Istraživanje je koncipirano kao studija preseka, koja obuhvata 50 pacijenata sa ACC pljuvačnih žlezda, muškog i ženskog pola, prosečne starosti 58 godina. U ovoj studiji koristili smo tehniku tkivnog mikroniza (TMA kalupi). Preseci sa TMA kalupa, debljine 5μm, bojeni su streptavidin-biotin imunohistohemijskom tehnikom pomoću primarnih antitela specifičnih za: Apaf-1, kaspazu 9 i kaspazu 3. Obojeni tkivni preseci su analizirani svetlosnim mikroskopom (Olympus tip BH-2). Na osnovu prikupljenih podataka formirana je baza podataka u –SPSS 18.0 koja je korišćena za dalju statističku obradu. Statistička analiza podataka obuhvatala je metode deskriptivne i analitičke (inferencijalne) statistike...Adenoid cystic carcinoma (ACC), is one of the most common malignant salivary gland tumors. It participates about 10-15% in salivary neoplasms. It is characterized by a slow growth, high incidence of the nerve invasion, rare regional metastasis, frequent local recurrence and slow development of progressive and relatively indolent distant metastases. The most important factors for the prognosis of this tumor are the clinical stage and histological appearance of the tumor, presence or absence of tumor tissue on the lines of resection, nerve invasion. Apoptosis is the regulatory mechanism of tissue homeostasis. Dysregulation of apoptosis is a relatively common feature of cancer cells. Depending on the stimuli, activation of apoptotic pathways may include: receptor (extrinsic) and mitochondrial (intrinsic) pathways. In the intrinsic apoptotic pathway, Apaf-1, initiator and effector caspases play an important role. Apaf-1 is a multi domain protein, that binds cytochrome c and procaspase 9, which leads to the formation of apoptosome. This complex proteolytically activates the initiator caspase-9. In the final stage of apoptosis the activated caspase 9 leads to activation of the effector caspase 3, whose entry into the nucleus initiates the degradation phase of apoptosis. Objective The aim of this study was to examine the immunohistochemical expression of Apaf- 1, caspases 9 and 3 in the ACC of the salivary glands, and to correlate the results with clinicopathological parameters in order to determine which of them is an independent prognostic factor. Materials and Methods The study was designed as a study of sections, which included 50 patients with ACC of the salivary glands, male and female, average age 58 years. In this study, we used the technique of tissue microarray (TMA moulds). Sections from the TMA mould, 5μm thick, were stained with the streptavidin-biotin immunohistochemical technique using primary antibodies specific for: Apaf-1, caspase 9 and caspase 3. Stained tissue sections were analyzed by the light microscope (Olympus type BH-2). Based on the data collected, the database was created in -SPSS 18.0, which was used for a further statistical analysis. The statistical data analysis included methods of descriptive and analytical (inferential) statistics..

Immunohistochemical Analysis and Prognostic Value of Markers of the Intrinsic Pathway of Apoptosis in Adenoid Cystic Carcinoma of the Salivary Glands

Adenoidni cistični karcinom (ACC), je jedan od najčešćih malignih tumora pljuvačnih žlezda. Učestvuje sa oko 10-15% kod salivarnih neoplazmi. Karakteriše ga spor rast, visoka incidenca ka invaziji nerva, retko regionalno metastaziranje, česti lokalni recidivi i spor razvoj progresivnih i relativno indolentnih udaljenih metastaza. Najznačajniji faktori za prognozu ovog tumora su klinički stadijum i histološki izgled tumora, prisustvo ili odsustvo tumorskog tkiva na linijama resekcije, invazija nerva. Apoptoza je regulatorni mehanizam tkivne homeostaze. Poremećaj regulacije apoptoze je relativno česta karakteristika maligne ćelije. U zavisnosti od stimulansa, putevi aktivacije apoptoze mogu biti: receptorski (spoljašnji) i mitohondrijski (unutrašnji) put. U unutrašnjem putu apoptoze značajnu ulogu igraju Apaf-1, inicijatorne i efektorne kaspaze. Apaf-1 je multidomenski protein, za koji se vezuje citohrom c i prokaspaza 9 što dovodi do formiranja apoptozoma. Ovaj kompleks proteolitički aktivira inicijatornu kaspazu-9. U završnoj fazi apoptoze, aktivirana kaspaza 9 dovodi do aktivacije efektorne kaspaze 3, čijim ulaskom u jedro započinje degradaciona faza apoptoze. Cilj U ovoj studiji cilj je bio ispitati imunohistohemijsku ekspresiju Apaf-1, kaspaze 9 i 3 u ACC pljuvačnih žlezda, i rezultate korelirati sa kliničko-patološkim parametrima kako bi se utvrdilo koji je od njih nezavistan prognostički parametar. Materijal i metode Istraživanje je koncipirano kao studija preseka, koja obuhvata 50 pacijenata sa ACC pljuvačnih žlezda, muškog i ženskog pola, prosečne starosti 58 godina. U ovoj studiji koristili smo tehniku tkivnog mikroniza (TMA kalupi). Preseci sa TMA kalupa, debljine 5μm, bojeni su streptavidin-biotin imunohistohemijskom tehnikom pomoću primarnih antitela specifičnih za: Apaf-1, kaspazu 9 i kaspazu 3. Obojeni tkivni preseci su analizirani svetlosnim mikroskopom (Olympus tip BH-2). Na osnovu prikupljenih podataka formirana je baza podataka u –SPSS 18.0 koja je korišćena za dalju statističku obradu. Statistička analiza podataka obuhvatala je metode deskriptivne i analitičke (inferencijalne) statistike...Adenoid cystic carcinoma (ACC), is one of the most common malignant salivary gland tumors. It participates about 10-15% in salivary neoplasms. It is characterized by a slow growth, high incidence of the nerve invasion, rare regional metastasis, frequent local recurrence and slow development of progressive and relatively indolent distant metastases. The most important factors for the prognosis of this tumor are the clinical stage and histological appearance of the tumor, presence or absence of tumor tissue on the lines of resection, nerve invasion. Apoptosis is the regulatory mechanism of tissue homeostasis. Dysregulation of apoptosis is a relatively common feature of cancer cells. Depending on the stimuli, activation of apoptotic pathways may include: receptor (extrinsic) and mitochondrial (intrinsic) pathways. In the intrinsic apoptotic pathway, Apaf-1, initiator and effector caspases play an important role. Apaf-1 is a multi domain protein, that binds cytochrome c and procaspase 9, which leads to the formation of apoptosome. This complex proteolytically activates the initiator caspase-9. In the final stage of apoptosis the activated caspase 9 leads to activation of the effector caspase 3, whose entry into the nucleus initiates the degradation phase of apoptosis. Objective The aim of this study was to examine the immunohistochemical expression of Apaf- 1, caspases 9 and 3 in the ACC of the salivary glands, and to correlate the results with clinicopathological parameters in order to determine which of them is an independent prognostic factor. Materials and Methods The study was designed as a study of sections, which included 50 patients with ACC of the salivary glands, male and female, average age 58 years. In this study, we used the technique of tissue microarray (TMA moulds). Sections from the TMA mould, 5μm thick, were stained with the streptavidin-biotin immunohistochemical technique using primary antibodies specific for: Apaf-1, caspase 9 and caspase 3. Stained tissue sections were analyzed by the light microscope (Olympus type BH-2). Based on the data collected, the database was created in -SPSS 18.0, which was used for a further statistical analysis. The statistical data analysis included methods of descriptive and analytical (inferential) statistics..

Effects of continuous-combined oral drospirenone- estradiol on blood pressure, body weight & lipid profile in early menopausal women

Background & objectives: Drospirenone (DRSP) is a progestin with antimineralocorticoid and anti-androgenic activity. When administered in combination with estradiol (E2), it relieves menopausal symptoms. The aim of this study was to evaluate the effects of DRSP/E2 on the reduction of cardiovascular risk factors in menopausal women with hypertension. Methods: A retrospective study was conducted at the Clinical Center of Serbia. The participants were 64 menopausal women [mean age=49.19±4.62 yr, mean body mass index (BMI)=25.08±2.94 kg/m2, mean amenorrhoeic period=2.48±2.46 yr]. The effects of DRSP 2 mg/E2 1 mg on 24 h blood pressure (BP) variability, heart rate (HR), anthropometric characteristics and hormone and lipid levels were evaluated in early menopausal women with previously untreated stage 1 hypertension. All analyses were carried out before and after six and 12 months of therapy. Results: DRSP/E2 significantly reduced daytime BP values during six and 12 months of therapy. The reductions in systolic and diastolic BPs ranged from about −4.50 to −8.50 and from −4.00 to −5.00 mmHg, respectively. There were no significant changes in nocturnal 24 h BPs. DRSP/E2 significantly reduced HR daytime and night-time during the follow up period. DRSP/E2 significantly lowered the BMI, concentrations of total cholesterol, low-density lipoprotein cholesterol and apolipoprotein B, while high-density lipoprotein cholesterol and apolipoprotein concentration increased. Interpretation & conclusions: Continuous long-term therapy with DRSP 2 mg/E2 1 mg significantly lowered 24 h systolic and diastolic BPs and reduced the risk of cardiovascular disease in early menopausal women with stage 1 hypertension. Timely initiated menopausal hormone therapy can have beneficial effects on BP and can reduce the incidence of cardiovascular disease in menopausal women

Expression of apoptotic protease activating factor-1 in adenoid cystic carcinoma of the salivary glands and its clinicopathological relevance

Background/Aim. Apoptotic protease-activating factor-1 (Apaf-1) is a key molecule in the intrinsic or mitochondrial pathway of apoptosis. Some pathological conditions, such as cancer, stroke, and neurodegenerative diseases, are the result of dysregulation in the intrinsic apoptosis pathway. The aim of this study was to analyze the immuno-histochemical expression of Apaf-1 in adenoid cystic carcinoma (ACC) cells of the salivary glands and its correlation with clinicopathological parameters of patients (gender, age, localization, histological type, and overall survival). Methods. Formalin-fixed, paraffin-embedded tissues of ACC of the salivary glands from 50 male and female patients with an average age of 58 years, were used for the study. We used the technique of tissue microarray (TMA blocks). Sections from the TMA mold, 5 μm thick, were stained with the streptavidin-biotin immunohistochemical technique using primary antibodies specific for Apaf-1 (Leica Biosystems, Newcastle, UK). Stained tissue sections were analyzed by the light microscope (Olympus type BH-2). Based on the data collected, the database was created in SPSS software v. 22.0 (SPSS Inc., Chicago, ILL, USA), which was used for further statistical analysis. The statistical data analysis included methods of descriptive and analytical (inferential) statistics. Results. The results of the immunohistochemical analysis of Apaf-1 expression in the samples of patients with ACC of the salivary glands were compared with the clinicopathological parameters of these patients. The immunohistochemical expression of Apaf-1 showed no statistical significance with regard to the patients’ gender (p = 0.552), age (p = 0.106), histological tumor type (p = 0.654), and localization of ACC in the salivary glands (p = 0.486). There was no statistically significant correlation observed between the overall survival of ACC patients and Apaf-1 expression in tumor cells (p = 0.340, Log-Rank test). Conclusion. With regard to ACC, Apaf-1 expression is not in correlation with clinicopathological parameters (gender, age, localization, histological tumor type, outcome of the disease, and overall survival). Therefore, we believe Apaf-1 cannot be regarded as an independent prognostic factor for course and outcome of ACC of the salivary glands

The Interactions of Magnesium Sulfate and Cromoglycate in a Rat Model of Orofacial Pain; The Role of Magnesium on Mast Cell Degranulation in Neuroinflammation

Mast cell degranulation impacts the development of pain and inflammation during tissue injury. We investigated the antinociceptive effect of a combination of cromoglycate and magnesium in the orofacial model of pain and the histological profile of the effect of magnesium in orofacial pain. In male Wistar rats, formalin (1.5%, 100 µL) was injected subcutaneously into the right upper lip of rats after cromoglycate and/or magnesium. Pain was measured as the total time spent on pain-related behavior. Toluidine blue staining was used to visualize mast cells under the light microscope. In the formalin test, in phase 1, magnesium antagonized the antinociceptive effect of cromoglycate, while in phase 2, it potentiated or inhibited its effect. Magnesium significantly reduced mast cell degranulation in the acute phase by about 23% and in the second phase by about 40%. Pearson’s coefficient did not show a significant correlation between mast cell degranulation and pain under treatment with magnesium. The cromoglycate–magnesium sulfate combination may prevent the development of inflammatory orofacial pain. The effect of a combination of cromoglycate–magnesium sulfate depends on the nature of the pain and the individual effects of the drugs. Magnesium reduced orofacial inflammation in the periphery, and this effect did not significantly contribute to its analgesic effect

Immunohistochemical expression of caspases 9 and 3 in adenoid cystic carcinoma of salivary glands and association with clinicopathological parameters

Purpose: Adenoid cystic carcinoma (ACC) is one of the most common malignant salivary gland tumors. It is characterized by a high rate of recurrence, perineural invasion and development of distant metastases many years after removal of the primary tumor. Disorders of the induction of apoptosis and its cascade reactions where caspases are involved may be significant in the pathogenesis of this tumor. Methods: The immunohistochemical expression of caspase 9 and caspase 3 was analyzed by tissue microarray (TMA) in 50 cases of ACC in relation with different clinicopathological parameters (gender, age, localization, histological type and overall survival). Results: Caspase 9 was expressed in the cytoplasm and nuclei of ACC tumor cells with varying degrees of staining intensity (1+, 6%; 2+, 54%, 3+, 40%). Comparison of caspase 9 expression in tumor cells with clinicopathological parameters (gender, age, localization, histological type and overall survival) showed no statistically significant difference except that the expression was more pronounced in females. Caspase 3 was expressed in the cytoplasm of tumor cells with varying degrees of staining intensity (1+, 22%; 2+, 36%; 3+, 42%). No correlation between the expression of caspase 3 and clinicopathological parameters was noticed. Conclusions: The expression of caspases 9 and 3 in ACC of the salivary glands can contribute in the better characterization of molecules involved in apoptosis of tumor cells