Heterogeneous vascular patterns in renal cell carcinomas.

The present study proposes a classification of renal cancer tumor blood vessels according to their morphology and maturation grade. We identified four vascular patterns: reticular, diffuse, fasciculated and trabecular. The reticular pattern was present in 63% of cases, being characterized by the predominance of mature CD34+/SMAct+ tumor vessels, highly interconnected. For this pattern, 74% of cases had vascular invasion, and a significant correlation was observed between tumor grade and immature state of tumor vessels (p = 0.022). The diffuse pattern was observed in 23% of cases and was characterized by non-interconnected vessels predominantly of mature CD34+/SMAct+ type and vascular invasion in 64% of cases. Only 8% of cases, had a fasciculate model of vessels distribution, all of them being of mature type, located in the connective axis of papillary renal tumors. For this pattern vascular invasion was found in 50% of cases. In 6% of cases a trabecular pattern was observed and the lowest rate of vascular invasion was registered. We defined here four distinct vascular patterns in renal cell carcinomas showing a strong impact on vascular invasion. A complete morphological and molecular characterization of tumor vessels would be beneficial in elucidating the mechanisms that underlie the ineffectiveness of antiangiogenic/antitumor therapies.

[Immunohistochemical study of the role of mast cells and macrophages in the process of angiogenesis in the atherosclerotic plaques in patients with metabolic syndrome].

OBJECTIVE: to analyze the histotopographic distribution of neogenic vessels, the degree of mast cell degranulation, the expression of markers of macrophages in different types of plaques in different stages and in different arterial vessels in patients with atherosclerosis and in those with metabolic syndrome associated with atherosclerosis and to establish the role of mast cells and macrophages in the development of stages of atherosclerosis along with their diagnostic and prognostic value. MATERIAL AND METHODS: Fragments of the thoracic and abdominal aorta, middle cerebral, carotid, renal, and iliac, and vertebral arteries from 34 persons who had died from atherosclerosis (n=17) and atherosclerotic complications due to metabolic syndrome (n=17) were examined. The investigators employed standard techniques, such as hematoxylin-eosin or orcein staining; silver impregnation. They used immunohistochemical staining with anti-mast cell tryptase (anti-MCT) for the determination of mast cells, the specific markers CD68 for macrophages, and CD105 (endoglin) for neogenic vessels. RESULTS: The immunohistochemical technique is effective in identifying mast cells, macrophages, and neogenic vessels in atherosclerotic plaques. They were found in many types of atherosclerotic plaques, advantium, and subendothelial layers in the immediate vicinity of the plaques. There was a statistical correlation between the types of plaques and clinical data, which is of importance in elucidating the specific features of the pathogenesis of atherosclerosis in patients with metabolic syndrome. CONCLUSION: CD105 is a sensitive marker for neogenic endothelial cells, an effective indicator of microvascular activation and proliferation in the atherosclerotic plaques. Neovascularization in the plaques frequently begins in the intima, progresses, and gives rise to their further destabilization. Anti-MCT staining used to reveal mast cells and CD68 for macrophages can elucidate the important patterns of development of atherosclerosis and its complications in patients with metabolic disturbances.

Circular anastomotic experimental fibrin sealant protection in deep colorectal anastomosis in pigs in a randomized 9-day survival study.

PURPOSE: The reported rate of clinically apparent anastomotic leakage (AL) in a low anterior resection of the rectum (LAR) (≤7 cm from the anal verge) using a circular double-stapled anastomosis (CDSA) without defunctioning stoma is up to 37.5 %. Since AL may result in life-threatening peritonitis, sepsis, and multiple organ failure, LAR and CDSA are regularly combined with defunctioning stoma. Accordingly, we now evaluated whether LAR and CDSA without defunctioning stoma but with extraluminal anastomotic application of an experimental fibrin sealant reduce the AL rate. This might prevent humans from defunctioning stoma increasing quality of life and decreasing surgical costs. METHODS: Forty 8-week-old pigs underwent LAR and CDSA in an end-to-end technique (descendo-rectostomy). Animals were randomized into a therapy and control group (gr.). The therapy gr. (n = 20) received an additional extraluminal circular application of an experimental fibrin sealant to the anastomosis. The objective was to assess the incidence of clinically apparent and non-clinically apparent leakage through the ninth postoperative day. Double-contrast barium CT radiographs of the colorectal region were performed on the ninth postoperative day or earlier, in case there were clinical signs of AL. All remaining animals were sacrificed on the ninth postoperative day and the anastomotic region was histopathologically analyzed. In case of earlier diagnosed AL, animals were sacrificed immediately. Blood samples were taken for complete blood count, chemistry, and coagulation profile prior to surgery and on the first, third, fifth, seventh, and ninth postoperative day. RESULTS: A circular extraluminal anastomotic application of an experimental fibrin protection decreased the rate of clinically and non-clinically apparent AL from 20 % (n = 4) in the control group to 5 % (n = 1) in the treatment group. Ulcerations were also observed in both gr. (control gr.-5 animals, therapy gr. -3 animals). All animals with AL showed necrosis surrounding the hole at the anastomoses. Three additional animals had a full wall defect at the anastomotic region that was blocked by the experimental fibrin sealant. The fibrin sealant was present at necropsy in all treated animals. CONCLUSION: Circular anastomotic protection with the experimental fibrin sealant blocked anastomotic full wall defects, preventing peritonitis and significantly reducing the AL rate from 25 to 5 %.

Mast cell phenotype in benign and malignant tumors of the prostate.

The molecular phenotypic heterogeneity of mast cells (MCs) makes them attractive as potential therapeutic targets in anti-cancer adjuvant therapy. Mast cell aggregations observed in tumors suggested their involvement in tumor pathogenesis. Despite several studies using mast cell tryptase, MCs' involvement in the progression of prostate tumors has not been demonstrated. The aim of our study was to identify and quantify the phenotypic heterogeneity of MCs in prostate lesions. Our study included 7 cases of normal prostate, 25 cases of benign epithelial hyperplasia and 64 cases of prostate carcinoma. MCs were immunohistochemically assessed using three markers: tryptase, chymase and CD117. Two immunophenotypes of MCs were identified in benign lesions: tryptase+/CD117+/chymase- and tryptase-/chymase+/CD117+, located in peritumoral areas. Intratumoral MC phenotype of malignant lesions was characterized by tryptase+/chymase+/CD117+, while in the peritumoral areas three different MCs phenotypes were identified: tryptase+/chymase+/CD117-, tryptase+/CD117+/chymase- and chymase+/CD117+/tryptase-. Our results suggest the correlation of chymase positive MCs of the peritumoral areas and CD117 positive MCs of the intratumoral areas with tumor grade.

Myxoid/round cell conjunctival liposarcoma. A case report.

PURPOSE: To present a rare case of conjunctival myxoid liposarcoma, subtype round cells, that had a seven years follow up. CLINICAL OBSERVATION: A 61-year-old female patient presents with a palpable, non-painful tumor, on the superior temporal bulbar conjunctiva of the right eye. The initial examination detects a fleshy tumor, orange in color, under the superior temporal bulbar conjunctiva, as well as two oval-shaped hyperpigmented conjunctival lesions, near the limbus at 10 o'clock, causing moderate blepharoptosis. Vision was normal, there was no diplopia, proptosis, afferent pupillary defect or lymphadenopathy; there was no orbital involvement in MRI. An isolated 15/15 mm tumor, with no connections with the eye socket, was excised. Histopathology revealed a poorly differentiated myxoid liposarcoma. Five recurrences occurred, of which four were treated by local excision and the last required exenteration. Repeat histopathology detects lipoblasts, small round cells, with immunohistochemistry positive for CD34, S100 and vimentin. The last two rapidly evolving and large recurrences, as well as pulmonary metastasis and finally death of the patient, underlined the aggressive character of round cell conjunctival liposarcoma. CONCLUSIONS: Conjunctival myxoid liposarcoma is characterized by numerous local recurrences, but the speed of the succession and volume of the recurrences may suggest a change in the underlying histopathological aspect, that is definitory for the therapeutical and prognostic approach of the case.

From basic lesions to a pathological staging of pulmonary fibrosis.

Idiopathic pulmonary fibrosis is a severe disease, with unpredictable evolution that frequently leads to respiratory failure and death, despite some progresses made in the field of therapy. Basically, the bad prognosis and failure of therapy are the consequence of the lack of data about the molecular events that have as result the extensive fibrosis. Although the basic lesions were defined many years ago, the pathological classification of pulmonary fibrosis is controversial. In the present work, we analyzed the prognostic impact of basic microscopic lesions on a possible new classification that could be related to the patient outcome. For this purpose, we have investigated 20 cases with idiopathic pulmonary fibrosis and samples of lung parenchyma were obtained by video assisted thoracoscopy. The specimens were processed by usual histological technique and sections were stained with Hematoxylin-Eosin, Masson's trichrome and Gordon-Sweet silver staining. There were evaluated the lung architecture, the chronic inflammatory infiltrate, macrophages and fibrosis. The distribution and severity of each parameter was converted into points and finally graded from I to IV, with corresponding score from 1 to 12. We found four cases with degree II, 12 with degree III, and four with degree IV. Our results support the hypothesis that the evaluation of basic lesions could be the basis for a more objective classification and staging of lung fibrosis and, possibly, a better prognostic method and, eventually, a predictor for the response to targeted therapy.

Behavior of the P1.HTR mastocytoma cell line implanted in the chorioallantoic membrane of chick embryos.

The P1.HTR cell line includes highly transfectable cells derived from P815 mastocytoma cells originating from mouse breast tissue. Despite its widespread use in immunogenic studies, no data are available about the behavior of P1.HTR cells in the chick embryo chorioallantoic membrane model. The objective of the present investigation was to study the effects of P1.HTR cells implanted on the chorioallantoic membrane of chick embryos. We inoculated P1.HTR cells into the previously prepared chick embryo chorioallantoic membrane and observed the early and late effects of these cells by stereomicroscopy, histochemistry and immunohistochemistry. A highly angiotropic and angiogenic effect occurred early after inoculation and a tumorigenic potential with the development of mastocytoma keeping well mast cells immunophenotype was detected later during the development. The P1.HTR mastocytoma cell line is a good tool for the development of the chick embryo chorioallantoic membrane mastocytoma model and also for other studies concerning the involvement of blood vessels. The chick embryo chorioallantoic membrane model of mastocytoma retains the mast cell immunophenotype under experimental conditions and could be used as an experimental tool for in vivo preliminary testing of antitumor and antivascular drugs.

Behavior of the P1.HTR mastocytoma cell line implanted in the chorioallantoic membrane of chick embryos

The P1.HTR cell line includes highly transfectable cells derived from P815 mastocytoma cells originating from mouse breast tissue. Despite its widespread use in immunogenic studies, no data are available about the behavior of P1.HTR cells in the chick embryo chorioallantoic membrane model. The objective of the present investigation was to study the effects of P1.HTR cells implanted on the chorioallantoic membrane of chick embryos. We inoculated P1.HTR cells into the previously prepared chick embryo chorioallantoic membrane and observed the early and late effects of these cells by stereomicroscopy, histochemistry and immunohistochemistry. A highly angiotropic and angiogenic effect occurred early after inoculation and a tumorigenic potential with the development of mastocytoma keeping well mast cells immunophenotype was detected later during the development. The P1.HTR mastocytoma cell line is a good tool for the development of the chick embryo chorioallantoic membrane mastocytoma model and also for other studies concerning the involvement of blood vessels. The chick embryo chorioallantoic membrane model of mastocytoma retains the mast cell immunophenotype under experimental conditions and could be used as an experimental tool for in vivo preliminary testing of antitumor and antivascular drugs.

The characterization of PDGFR-alpha and PDGFR-beta expression in malignant non-Hodgkin lymphoma.

PDGF receptors play an important role in tumor progression as being part of a group of receptors that are expressed along the membrane of tumor cells. The aim of this study was to evaluate the PDGF receptor expression in follicular and diffuse forms of non-Hodgkin malignant lymphoma. We evaluated 38 biopsy fragments from patients diagnosed with malignant non-Hodgkin lymphoma. We noticed the distribution of PDGFR-alpha and -beta in tumor cells and the immunoreactivity was quantified as the percentage of positive tumor cells. We noticed the presence of score 3, more than 30% of tumor cells positive, for PDGFR-alpha and PDGFR-beta in 50% and 75% cases of follicular non-Hodgkin lymphoma. For diffuse malignant non-Hodgkin lymphomas, score 3 was noted in 23.52% of cases for PDGFR-alpha and 35.29% of cases for PDGFR-beta. This may represent an important therapeutic target in patients who do not respond to conventional therapy, but further research is needed for a careful evaluation of benefits and side effects of PDGFR inhibitors.

A short story of Victor Hensen and a cell of the internal ear.

Cells of Hensen of the internal ear are known from more than 100 years and investigations on their function(s) are still waiting for an answer. They were first described by the German scientist from Kiel, Christian Andreas Victor Hensen. This short historical review gives some details about the life, scientific activity and perspectives opened by the work of Victor Hensen.

Prox 1, VEGF-C and VEGFR3 expression during cervical neoplasia progression as evidence of an early lymphangiogenic switch.

Prox1 is a key regulator of lymphatic endothelial cell commitment during embryonic development. No correlations between Prox1 and VEGF-C/VEGFR3 expression in cervical cancer has been done until now. The aim of the present study was to evaluate the peculiarities of Prox1, VEGF-C and VEGFR3 expression during uterine cervix neoplasia progression. Material and methods. One hundred and four specimens taken from women with macroscopically detectable lesions were classified by histopathology and analyzed by immunohistochemistry for Prox1, VEGFR3 and VEGF-C expression. Results. The presence of Prox1 nuclear expression was detected starting from CIN2 and CIN3 lesions to microinvasive carcinoma, in the nuclei of lymphatic and venous endothelial cells and scattered stromal cells. All Prox1 positive lymphatic vessels were positive for VEGFR3. A significant correlation was found between expression of VEGF-C in tumor cells and nuclear density of Prox1 positive lymphatic cells (p=0.044). Conclusion. The commitment of Prox1 positive cells through a lymphatic lineage is an early event for cervical neoplastic progression, being present starting with intraepithelial cervical lesions, and is strongly associated with VEGFR3 and VEGF-C expression. These findings suggest an early active lymphangiogenesis during cervical neoplasia progression and explain, in part, the early presence of lymph node metastasis in cervical cancer. By the detection of Prox1 expression in lymphatic and venous endothelial cells, and also in stromal cells, it has been suggested that there are at least three different mechanism of lymph vessel development during cervical neoplasia progression.

The immunohistochemical expression of endocrine gland-derived-VEGF (EG-VEGF) as a prognostic marker in ovarian cancer.

Ovarian cancer-related angiogenesis is a complex process orchestrated by many positive and negative regulators. Many growth factors are involved in the development of the tumor-associated vasculature, and from these, endocrine gland-derived vascular endothelial growth factor (EG-VEGF) seems to play a crucial role. EG-VEGF is the first organ-specific angiogenic factor and its effects are restricted to the endothelial cells of the endocrine glands. Although EG-VEGF was detected in both normal and neoplastic ovaries, its clinical significance remains controversial. In the present study, we analyzed 30 patients with epithelial ovarian cancer, and the immunohistochemical expression of EG-VEGF was compared with the conventional clinico-pathological parameters of prognosis. Neoplastic cells of the ovarian carcinoma expressed EG-VEGF in 73.33% of the cases, as a cytoplasmic granular product of reaction. We found a strong correlation between the expression of EG-VEGF at protein level and tumor stage, grade, and microscopic type. The expression of EG-VEGF was found in patients with stage III and IV, but not in stage II. The majority of serous adenocarcinoma, half of the cases with clear cell carcinoma and two cases with endometrioid carcinoma showed definite expression in tumor cells. No positive reaction was found in the cases with mucinous carcinoma. Our results showed that EG-VEGF expression is an indicator not only of the advanced stage, but also of ovarian cancer progression. Based on these data, we concluded that EG-VEGF expression in tumor cells of the epithelial ovarian cancer is a good marker of unfavorable prognosis and could be an attractive therapeutic target in patients with advanced-stage tumors, refractory conventional chemotherapy.

Clinical, morphological and immunohistochemical characterization of a recurrent B1 type thymoma.

Type B1 thymoma is widely accepted as a tumor with a non-aggressive behavior even in advanced stage. Most of these tumors are classified as Masaoka stage I or II. They rarely relapse or metastasize and the surgical treatment is considered curative. We have investigated a case of thymoma type B1, which relapsed 13 months after the primary tumor was excised. The patient was diagnosed with a local tumor recurrence after investigations due to the worsening of clinical symptoms of myasthenia gravis (MG). The therapy management of such cases is debatable and protocols not yet approved. For this reason, we have analyzed different clinical, morphological and immunohistochemical characteristics that may be considered as prognostic factors for a more aggressive behavior of such tumors. We have identified some morphologic characteristics rarely seen in this type of thymoma but none considered of prognostic value. In addition, we investigated some possible immunohistochemical markers that are generally associated with a more aggressive clinical outcome in different malignant tumors and thymic epithelial tumors. Among these markers, only p53 was positive and may be useful to predict a more aggressive evolution. In summary, probably the more appropriate approach of the patient is the clinical follow-up together with treatment of the clinical symptoms of myasthenia gravis.

Microvascular density in non-Hodgkin B-cell lymphomas measured using digital morphometry.

INTRODUCTION: The growth of solid tumors requires the development of microvessels, therefore tumor expansion depends on angiogenesis. Microvessels provide nutrients and oxygen and remove catabolytic substances, while endothelial cells produce growth factors for tumor cells in a paracrine fashion. The microvascular component of a tumor also plays a role in the metastatic capacity of the tumor, enabling the tumor cells to spread to distant locations by providing a large endothelial surface. AIM: The purpose of this study was to review the literature about angiogenesis regarding malignant lymphomas and to perform basic measurements by means of digital morphometric methods in large B-cell lymphomas and follicular lymphomas. MATERIALS AND METHODS: After thorough analyzing currently available assessment methods, we performed angiogenesis assessment on 19 randomly selected cases, from paraffin-embedded specimens using digital morphometry. We used immunohistochemistry and the CD34 antigen to mark microvessels. We measured average vascular diameter and a previously successfully applied digital morphometric method to quantify the extent of endothelial area. RESULTS: According to literature data, our knowledge and understanding of angiogenesis grew rapidly from early studies such as Folkman's classic paper. Many studies showed that angiogenesis plays a key role in the biology of tumors and therefore the study of angiogenesis might open new therapeutic possibilities. There have been many studies of angiogenesis in malignant lymphomas, however not as many articles as in other tumor types. Our morphometric studies showed there are statistically significant differences between diffuse large cell lymphoma (DLBCL) and follicular lymphoma (FL) regarding average vascular diameter and that high grade lymphomas tend to have a greater CD34+ endothelial area.

Morphological study of bronchial mucosa in the chronic obstructive pulmonary disease under the influence of therapeutic algorithm.

OBJECTIVES: Immunohistochemical evaluation of the effectiveness of bronchodilator treatment in patients with chronic obstructive pulmonary disease (COPD). MATERIALS AND METHODS: There have been examined bronchial mucosa biopsies taken endoscopically from 18 patients with obstructive pulmonary disease. The biopsies were fixed in 4% buffered formalin for 24-48 hours and paraffin inclusion was made using the standard technique. For each biopsy, there were performed 10 serial sections with a thickness of 5 µm. The sections were stained using morphological, histochemical and immunohistochemical methods. At three of the cases, the paraffin blocks were reconverted for the electron microscopy study, in order to assess subcellular details, with special reference to "target" cells involved in local immune response. Morphohistochemical and immunohistochemical analysis was effectuated on biopsies removed before and after the treatment with bronchodilators. RESULTS: The analysis of the biopsies removed before treatment revealed the following aspects: degenerative alterations of the surface epithelium, loss of ciliary differentiation, absence of caliciform cells, hyperexfoliation, formation of pseudopapillary structures, degenerative lesions of the glands, mucoid and oncocytary metaplasia, stasis in the dilated blood vessels, partly hyalinized wall, multiforme chronic inflammatory infiltrate, myofibroblasts in the depth of lamina propria; argyrophilic basement membrane, fragmentation and lysis of elastic fibers, degranulated mast cells associated with inflammatory infiltrate, with electron-dense typical granules, inflammatory infiltrate with CD20 positive B-lymphocytes, arranged perivasculary and in the vicinity of the basement membrane; rare positive CD4 T-lymphocytes; reduced number of plasma cells. After treatment we found the following aspects: partial or complete regeneration of the covering epithelium, with the presence of cilia cells and occasionally of caliciform cells; remaining myofibroblastic reaction in the lamina propria; increased number of mast cells with minimal or no degranulation; immature, lamelled mast cells. CONCLUSIONS: The application of management principles in group therapy study was done by the study which aims to demonstrate the beneficial role in COPD therapy of combining a ß2-agonist with an anticholinergic, obtaining in this way an additional bronchodilator effect, compared with the one obtained by administrating bronchodilators of type ß2 agonists. Deepening the molecular and cellular mechanisms of COPD can lead to more effective methods for early detection of disease, pharmacotherapy targeted and effective conduct exacerbations.

Heterogeneity of malignant non-Hodgkin lymphoma-associated blood vessels.

The present study described for the first time a high heterogeneity of blood vessels in non-Hodgkin lymphomas (nHL). The tumor blood vessels were highlighted with CD105÷smooth muscle actin (SMA) and CD34/SMA double immunostaining. For both follicular and diffuse types of lymphomas, more than 85% of CD34/SMA positive vessels were of immature and intermediate type. A percent of 96.54 from CD105/SMA assessed blood vessels were of activated and mature activated types with high expression of CD105 on endothelial cells of newly formed blood vessels. Our results suggest that these types of vessels are potential therapeutic targets for antivascular therapy.

Correlation between histopathological form and the degree of neuroendocrine differentiations in prostate cancer.

Prostate cancer (PCa) is the most frequent neoplasic condition in males, but only 64-65% of the cases are sensitive to hormone therapy. The aim of this study was to investigate the neuroendocrine component of the prostatic carcinoma, in relation to the histopathological form and the degree of differentiation. Biopsies were obtained through transurethral resection, from 82 patients with prostate cancer. In order to assess the histopathological form and the Gleason score, one section from each case was stained with Hematoxylin-Eosin. Additional sections were stained with chromogranin A. We considered neuroendocrine cell hyperplasia to have a higher value than that observed in benign prostatic hyperplasia (BPH) and normal prostate (over three neuroendocrine cells÷gland). The quantification of neuroendocrine differentiation (NED) has been significant; the reaction was considered to be weak (2-10% neuroendocrine cells), moderate (10-20%) and intense (over 50%). Cells positive for chromogranin A have been identified in all the cases, but a larger number than that registered in normal tissue has been noted in 59 patients (71.95%). In most of the cases, the neuroendocrine cells have been distributed in small groups among the neoplasic cells, and rarely isolated. In two cases of small cell carcinoma most of the tumoral cells have been positive for chromogranin A. In conclusion, the study of neuroendocrine differentiation in patients with prostatic carcinoma revealed hyperplasia of positive chromogranin A cells, in 71.95% of cases. Neuroendocrine prostatic differentiation is correlated with the advanced stage of evolution and possibly with the resistance to hormonal treatment.

Atypical variant of lichen planus mimicking normal skin histology.

We present the case of a 69-year-old Caucasian and non-smoker patient with erythematous, itchy, violaceous lesions on the ankles, wrists, lower legs, forearms and trunk developed within 15 months. Her condition was diagnosed as prurigo and she was treated for a long period of time with antihistaminic drugs, with no resolution of lesions. In October 2008, she presented to a Private Practice of Dermatology in Timisoara. The dermatologist noticed the development of violaceous lesions on her trunk. The patient had similar lesions on the ankles, wrists, lower legs, and forearms for the last eight months. At physical examination many hyperpigmented, 1 to 6 cm, thin plaques were present on the flanks, shoulders, and infra-mammary area. There was no vaginal involvement. This eruption had a good response to topical glucocorticoids, but recurred multiple times after discontinuation of treatment. A biopsy specimen showed some histopathological features of lichen planus.

Conventional examination versus immunohistochemistry in the prediction of hormone profile of pituitary adenomas. An analysis on 142 cases.

The large majority of neoplasms located in the sella turcica are benign pituitary adenomas derived from cells of the adenohypophysis. Pituitary adenomas represent the third most common primary intracranial tumor in neurosurgical practice, outnumbered only by gliomas and meningiomas. Their biology of pituitary adenomas is complex and they can cause a variety of endocrine syndromes and disorders, based on hormone profile secreted by proliferating cells. The aim of this study was to evaluate the routine conventional methods and highly specific immunohistochemical methods in order to accurately predict the type of hormone secretion. Our study was investigated 142 cases admitted with pituitary adenomas and treated by open surgery. Sections from each case were stained with routine Hematoxylin and Eosin method for histopathologic evaluation. Immunohistochemistry was performed on additional slides in order to detect specific pituitary adenomas. Based on the immunohistochemical profile of pituitary adenomas we found following results: 37 growth hormone (GH)-secreting adenomas, 23 prolactin (PRL)-secreting adenomas, 15 mixed GH-cell/PRL-cell adenomas, three mammosomatotroph cell adenomas, two adrenocorticotrophic hormone (ACTH)-secreting adenomas, one thyrotrophic cell adenoma, 18 gonadotroph adenomas, 30 null cell adenomas and 13 plurihormonal adenomas. No correlation was found between conventional features and the immunohistochemical profile. The immunohistochemical profile of functional pituitary adenomas is mandatory for a correct diagnosis. We revealed that staining characteristics of the tumor cells, such as acidophilic, basophilic or chromophobe are nowadays outdated as main principle of classification, because they not identify specific adenoma types.

MCF7-MSC co-culture assay: approach to assess the co-operation between MCF-7s and MSCs in tumor-induced angiogenesis.

The multipotent stromal cells (MSCs) exhibit a broad differentiation potential. MSCs might participate in neovascularization through their ability to migrate and generate capillary-like structures. These processes were shown to be modulated by tumor angiogenic factors, such as Vascular Endothelial Growth Factor (VEGF). The aim of our study was to define the way the MCF-7 cell line (MCF-7s) influenced the MSCs' recruitment for the tumor-induced angiogenesis, and to assess the role of VEGF in this process. We tested the chemotactic potential of plasma or VEGF, but also of MCF-7s or their conditioned medium (CM) in the MSCs' transmigration. We compared the migratory potential of MSCs, MSCEs (MSCs cultured in endothelial cell growth medium) and HUVECs. Recombinant VEGF has been shown to chemoattract MSCs, although to a lesser degree than plasma or serum containing medium alone. Moreover, it changed the MSCs' morphology, stimulating the appearance of longer and thinner prolongations as compared to plasma. MCF-7s or their CM both directly induced migration of MSCs. Surprisingly, CM augmented with MCF-7s attracted less cells than the control medium itself, but CM augmented or not with MCF-7s changed the morphology of MSCs in a manner similar to VEGF. The migratory behavior of the MSCEs was comparable to that of HUVECs, while their morphology could be considered intermediate between MSCs and HUVECs, as they developed shorter prolongations than MSCs, but much longer than HUVECs in the corresponding wells. In conclusion, both tumor cells and VEGF alter the migration behavior of MSCs in a transmigration model, indicating a role of tumor cell-derived VEGF to modulate the recruitment of MSCs into sites of angiogenesis.