Anatomical organization of the lateral cervical nucleus in Artiodactyls

The presence of the lateral cervical nucleus (LCN) in different mammals, including humans, has been established in a number of anatomical research works. The LCN receives its afferent inputs from the spinocervical tract, and conveys this somatosensory information to the various brain areas, especially the thalamus. In the present study, the organization of the calf and pig LCN was examined through the use of thionine staining and immunohistochemical methods combined with morphometrical analyses. Specifically, the localization of calbindin-D28k (CB-D28k) and neuronal nitric oxide synthase (nNOS) in the LCN was investigated using the immunoperoxidase method. Calf and pig LCN appear as a clearly defined column of gray matter located in the three cranial segments of the cervical spinal cord. Thionine staining shows that polygonal neurons represent the main cell type in both species. The calf and pig LCN contained CB-D28k-immunoreactive (IR) neurons of varying sizes. Large neurons are probably involved in the generation of the cervicothalamic pathway. Small CB-D28k-IR neurons, on the other hand, could act as local interneurons. The immunoreactivity for nNOS was found to be mainly located in thin neuronal processes that could represent the terminal axonal portion of nNOS-IR found in laminae III e IV. This evidence suggests that nitric oxide (NO) could modulate the synaptic activity of the glutamatergic spinocervical tracts. These findings suggest that the LCN of Artiodactyls might play an important role in the transmission of somatosensory information from the spinal cord to the higher centers of the brain.

Anatomical Organization of the Lateral Cervical Nucleus in Artiodactyls

The presence of the lateral cervical nucleus (LCN) in different mammals, including humans, has been established in a number of anatomical research works. The LCN receives its afferent inputs from the spinocervical tract, and conveys this somatosensory information to the various brain areas, especially the thalamus. In the present study, the organization of the calf and pig LCN was examined through the use of thionine staining and immunohistochemical methods combined with morphometrical analyses. Specifically, the localization of calbindin-D28k (CB-D28k) and neuronal nitric oxide synthase (nNOS) in the LCN was investigated using the immunoperoxidase method. Calf and pig LCN appear as a clearly defined column of gray matter located in the three cranial segments of the cervical spinal cord. Thionine staining shows that polygonal neurons represent the main cell type in both species. The calf and pig LCN contained CB-D28k-immunoreactive (IR) neurons of varying sizes. Large neurons are probably involved in the generation of the cervicothalamic pathway. Small CB-D28k-IR neurons, on the other hand, could act as local interneurons. The immunoreactivity for nNOS was found to be mainly located in thin neuronal processes that could represent the terminal axonal portion of nNOS-IR found in laminae III e IV. This evidence suggests that nitric oxide (NO) could modulate the synaptic activity of the glutamatergic spinocervical tracts. These findings suggest that the LCN of Artiodactyls might play an important role in the transmission of somatosensory information from the spinal cord to the higher centers of the brain.

Histological and immunohistochemical evaluation of epithelial and mesenchymal tumors of psittacines

This study describes histological and immunohistochemical evaluation of 50 epithelial and mesenchymal tumors of psittacines that were found in the archives of the pathology departments of veterinary faculties at Utrecht University and Istanbul University Cerrahpasa. In each case, cellular morphological features, tumoral pattern and mitotic activity were evaluated histologically, and immunohistochemistry was performed by the streptavidin-biotin immunoperoxidase method using a panel of commercially available antibodies raised against pan-cytokeratin, vimentin, muscle-specific actin, desmin, smooth muscle actin and Factor VIII/von Willebrand Factor. The histological evaluation revealed that 30 epithelial tumors were squamous cell carcinoma (n: 20), renal adenocarcinoma (n: 3), basal cell carcinoma (n: 2), abdominal carcinoma (n: 2), uropygial gland carcinoma (n: 1), cloacal carcinoma (n: 1) and cholangiocarcinoma (n: 1). Out of the 50 tumors, 13 had a mesenchymal origin and were diagnosed as fibrosarcoma (n: 4), hemangioma (n: 3), hemangiosarcoma (n: 4) and leiomyosarcoma (n: 2). Histological diagnosis could not be made for 7 tumors, and they were evaluated as undifferentiated malignant tumors (UDMT). Two tumors diagnosed as fibrosarcoma and three tumors diagnosed as UDMT showed strong SMA staining and were re-diagnosed as leiomyosarcoma. In view of the IHC findings, two other UDMTs were re-diagnosed as fibrosarcoma and anaplastic carcinoma. Although only a limited number of antibodies were used in the control panel, they were useful in establishing a definite diagnosis in some cases

The Expression of Progesterone Receptors in Meningiomas of Different Grades

Background: Meningiomas are slow growing intracranial and intraspinal neoplasms with a tendency to recur locally. WHO grades them as I (benign), II (atypical) and III (anaplastic) in order of their increasing aggressiveness, based on histological parameters and brain parenchymal invasion. Progesterone receptors (PR) are more prevalent amongst the lower grade meningiomas. The objective of this study was to determine the immunohistochemical expression of progesterone receptors in meningiomas of different grades.Material and Methods: A total of 100 cases were selected over a period of 2.5 years. Three to five microns’ thick sections stained with Hematoxylin and Eosin were examined microscopically by a team of two Histopathologists and graded into grades I, II and III, according to 2016 WHO classification criteria. Another section of the original tumor was stained with progesterone receptor antibody using the conventional immunoperoxidase method. Stained slides were than examined by the same team of Histopathologists and declared positive (if nuclear staining was observed in more than 10% of tumor cells) or negative. Statistical analysis was done using SPSS version 21.Results: Out of a total of 100 cases of meningioma, there were 79 cases of benign/typical WHO grade I, 15 cases of atypical/ WHO grade II and 6 cases of anaplastic/ WHO grade III tumor. PR status was positive in 89.8 % (71/79) of grade I meningiomas and 46.6 % (7/15) of grade II/Atypical meningiomas. The 06 cases of Anaplastic/WHO grade III tumors were negative for PR. There was a higher prevalence of Progesterone receptors in female patients (89.8%; 53/59) as compared to male meningioma patients (60.9%; 25/41).Conclusion: We observed a decreased expression of progesterone receptor in higher grades of meningioma in this study. It is an effort to explore conservative treatment options for inoperable lesions, as anti-progesterone therapy may hold a promise as a new treatment option in the near future.

Investigations of Pathological Immunohistochemical and Immunocytochemical Findings in Natural Infection with Mycoplasma gallisepticum in Laying Hens

Background: Mycoplasmosis is an infectious disease caused by Mycoplasma gallisepticum (MG), usually seen in the respiratory system of chickens, chick and turkeys, that causing great economic loss. The disease is characterized by respiratory system lesions such as sinusitis, tracheitis, airsacculitis, pneumonia and other symptoms such as loss of yield, arthritis, tenosynovitis. In this study, it was aimed to investigate diagnose of the disease by pathologic and molecular techniques in hens that naturally infected with MG as well as the usability of immunocytochemical (ICC) method in diagnose of the disease.Materials, Methods & Results: For this purpose, 98 hens were collected from 10 different coops that serologically positive. After necropsy, routine pathological procedures were performed to samples taken from nose, sinus, larynx, trachea, lung and air sacs. Scraping samples taken from lungs and tracheas were evaluated by ICC. Immunohistochemical (IHC) staining was performed to samples taken from nose, sinus, larynx, trachea, lung and air sacs. Indirect immunoperoxidase method was applied in the both IHC and ICC staining. Rabbit polyclonal anti MG antibody was used as primer antibody in the IHC and ICC staining. Additionally, culture and PCR techniques were applied to tracheas of all hens for MG. The GPO3 and MGSO genes were made for PCR analysis.In the tracheal examinations, 23 cases were positive for PCR, 17 cases ICC positive, 16 cases IHC positive and 10 culture samples found positive. All of culture positive cases were also positive for other three methods. When findings in all organs were evaluated, in 37 cases were detected positive by IHC (38%) and 23 cases were positive by ICC (23.5%). In the IHC positive cases, the first order was trachea in 16 cases followed by in 11 cases in sinus, in 8 cases in lung, in 6 cases air sac and 4 cases in nose, respectively. In 8 cases, IHC positivity was found in at least two organs. IHC positivity was detected in the nose, sinus and tracheal epithelia as well as in the macrophages within subepithelial lymphoid infiltration, vascular walls and endothelium. As the disease became chronic, it was found that the agents were seen more in the lymphoid tissue than the epithelium. In ICC staining positivity was found in 17 cases in the trachea and 11 cases in the lung. There were only 5 cases positive by ICC in both organs.Discussion: Clinical and pathological findings as well as serological, microbiological, molecular techniques and immunohistochemical methods are to be important methods in the diagnosis of the disease. While the culture results are shown as the gold standard in diagnosis of the disease, it is possible to obtain the results in the earliest 7-10 days in cultures and at least 20 days must be passed in order to say a cultural negative. In addition, in the field studies, it mentioned the use of vaccines, antibiotics and protective drugs affected the results of microbiology and serology; the importance of using techniques such as IHC and PCR for the diagnosis of the causative agents. The results of the present study indicate that the most important organ in the diagnosis of the disease is the trachea, and the most effective method is PCR followed by IHC and ICC methods. It was concluded that the results of ICC staining close to IHC staining, and ICC could be used for diagnostic purposes in positive reactions obtained from the tracheas or the other organs.

Arteriovenous forms of congenital vascular malformations of extremities: pathogenetic ground for modern approaches in treatment

Purpose of the study. Improvement of the results of treatment of patients with arteriovenous forms (AVF) of congenital vascular malformations (CVM) of the extremities based on the development of the classification scheme of CVM and the study of proliferative activity of the pathology.Materials and methods. The clinical data of 155 patients with AVF of CVM of extremities were analyzed in terms of observation from 1 month to 10 years. Patients of sex of men there were 65 (42%), women – 90 (58%), their correlation – 1 : 1,4; the average age was about 25,1 ± 10,4, children's age group – 53 (34%). The division into clinical-anatomical forms (11 groups) was carried out on the basis of the «working» classification scheme «VASC + T». Expression proliferation markers VEGF and KI-67 woos study were performed by immunoperoxidase method with additional hematoxylin staining.Results and discussion. Pathomorphological (66; 45,8%) and immunohistochemical studies (10; 7%) revealed the presence of proliferative activity of angiomatous tissues and degenerative changes in the walls of the vessels both due to developmental defects and disorders of hemodynamics, while the source of progression of the AVF of CVM there are vessels of the microvasculature.Conclusions. The application of the CVM classification scheme and the step-by-step complex treatment approach, depending on the clinical and anatomical AVF of CVM, led to the absence of major postoperative complications. This tactic made it possible to achieve satisfactory long-term results in 136 (94,4%) cases. Keywords: congenital vascular malformation, arteriovenous forms, proliferative activity, classification, complex treatment.

The Autophagy Creates Balance Mechanism between Apoptosis and Necroptosis: A Scorpion (Mesobuthus Nigrocinctus) Envenomation-Anti-Venom Administration Modelling in Mice

In the study, we aimed to show the role of autophagy which acting as a seesaw between apoptosis and necroptosis increasing and decreasing changes in certain vital organs under effects of envenomation by Mesobuthus nigrocinctus and anti-venom administration in mice. In group design, we previously classified totally 42 Albino mice into two main categories including 1st, 3th and 6th hours of envenomation and anti-venom administrated animals (at dosages of 20LD50, 30LD50 and 40LD50) before standard envenomation procudure. These were sub-divided into six groups including envenomed and anti-venom administrated mice (n = 6 at each one) were studied as well as not being evenomed and not anti-venom-administrated mice (n = 6) were used as control group (n = 6). At the end of 24 h after envenomation, all animals were sacrificed. Liver, kidneys, spleen, heart and lungs tissues were collected from all groups. After routine histopathological examination, expressions of mTOR as an autophagy activator, expressions of RIPK3 as a necroptosis activator, and caspase-3, caspase-9 as the markers of apoptotic cell death signals were evaluated by immunoperoxidase method in addition to DNA in-situ fragmentations by TUNEL method in aforementioned tissues. In envenomed groups, caspases and TUNEL reactions were low after envenomation in contrast to high RIPK3 expressions. mTOR expressions were remained at stable. In anti-venom administration groups, mTOR and RIPK3 expressions were more decreased in contrast to higher caspases exspressions and TUNEL expressions. In conclusion, we think that the scorpion envenomation drift solely the cells to necroptosis. And, under presence of anti-venom supply, the decreased mTOR expressions triggered autophagy and dependently that apoptosis was developed instead of necroptosis. We believed that this venom-antivenom administration study is a usefull role-model for better understanding of switch effect of autophagy between apoptosis and necroptosis.

Features of the nuclear steroid receptors expression in endometrium of women with failed in vitro attempts in history

Structural and functional maturity of the endometrium is formed in a condition of dynamic fluctuations of ovarian steroid hormones – estrogens and progesterone during the menstrual cycle. The objective: of the research was to determine the reactivity of estrogene core receptors-б and progesterone core receptors in endometrial stroma and glands. Patients and methods. Endometrial samples were obtained during hysteroscopy in 42 patients with infertility on day LH + 6 – LH + 11 of the menstrual cycle. The control group (CG) consisted of 18 healthy women who had at least one physiological pregnancy that resulted in the birth of a healthy child. Receptivity of the endometrium to the hormones was studied by immunoperoxidase method using mouse MAbs for estrogen receptors-a (ER-a) (clone ID5, «DAKO», Denmark) and progesteron receptor (RP) (clone A6, «DAKO», Denmark). The color intensity was assessed by tree-point system. The expression levelі of ER-a and RP were set by calculating semiquantitative index: IRS = SI ґ PP, where IRS – immunoreactivity index; SI – optical intensity staining; PP – the percentage of positively stained nuclei. Results. Normal ER-a and RP immunoreactivity was observed in 8 (19.05%) women with failed in vitro fertilization attempts in history, total reduction of ER-a and RP immunoreactivity (medium and severe) in glands and stroma was observed in 6 (14.29%) of infertile patients. Normal levels of ER-a and RP immunoreactivity were observed among KG women. Noteworthy is the presence of comorbidity. So in 4 of the six cases of endometrial polyp appeared on the background of chronic endometritis. In 4 women diagnosed with simple hyperplasia without atypia the defective rejection of the endometrium from the previous cycle and the incompatibility of endometrium structure to the day of cycle were found. In the group of infertile women high reactivity of receptors was observed only in 8 (19.05%) biopsies. Deviation of the morphological structure of the endometrium in this subgroup was observed significantly more often than in KG: incompatibility of morphological endometrial structure to the day of menstrual cycle was found in 12.50% of cases, simple hyperplasia without atypia – of 55.00%, endometrial polyp – 12.50%, chronic endometritis – 25.00%. Conclusions. The change of immunoreactivity of estrogen receptor-б and progesterone plays an important role in the formation of implantation failure in women with unsuccessful in vitro fertilization attempts. The obtained data confirm the hypothesis of multiple factors of development regulation of the endometrium in natural cycles and in IVF cycles. High frequency of structural and functional inferiority of the endometrium is the indication for in-depth study. Correction of the endometrial condition need to be done before pregnancy. Key words: infertility, failed in vitro fertilization attemp, immunoreactivity, receptors estrogen-a, progesterone receptors.

HISTOLOGICAL CHARACTERISTICS OF THE QUILTY EFFECT

Aim. Identifi cation of possible histological differences of the Quilty effect in acute rejection and its absence as well as studying the proliferation of blood and lymph vessels in the area of Quilty damage.Materials and methods. 883 endomyocardial biopsy materials from 352 patients were studied. Histological sections were stained with hematoxylin and eosin, Masson method; the endothelium of lymphatic vessels was stained with immunoperoxidase method using a marker D2-40.Results. The Quilty effect was observed both in acute rejection and inits absence. In the majority of cases the Quilty effect was of type «A» and it was combined with acute rejection. The Quilty effect of type «B» has been mostly in the G2R. Acute rejection is characterized by diffuse form of endocardium lymphoid infi ltration. Follicular form resembles lymphoid organ tissue. Different types and forms of the Quilty effect may be combined in the same biopsy. Proliferation of blood vessels presents in the area of the Quilty effect. Lymphatic vessels are missing in endocardium and in the area of the Quilty effect. They could be found only in the myocardium of endomiocardial biopsy.Conclusion. Diffuse lymphoid infi ltration of the endocardium is a characteristic feature of acute rejection of the transplanted heart. Follicular form of the Quilty effect is similar to lymphoid tissue whose role requires further study using immunohistochemical methods.