The significance of immune microenvironment in patients with endometriosis.

Endometriosis represents an estrogen-dependent disease of the female reproductive system and intra- and extraperitoneal regions, with chronic feature. Currently, immune cells, such as macrophages and lymphocytes, are considered to play a pivotal role in angiogenesis and invasion of endometriotic cells through matrix remodeling. Additionally, various studies have revealed the role of E-cadherin, ß-catenin, along with steroid hormone receptors in endometriosis development. In this context, our study aimed to analyze the relationship between the cellular immune profile and E-cadherin, ß-catenin, estrogen receptor alpha (ERα), and progesterone receptor (PR) immunoexpression in endometriosis tissues, along with an analysis of the possible association between serological parameters and immunohistochemical (IHC) markers. The study included 53 patients diagnosed with ovarian or cutaneous abdominal wall endometriosis, which have been investigated by routine histology, immunohistochemistry, and serum analysis. The IHC exam showed an increased density of cluster of differentiation (CD)4+ T-cells, CD8+ T-cells, and CD68+ macrophages, along with variable increased expressions of E-cadherin, ß-catenin, ERα, and PR. Statistical analysis revealed an intense positive correlation between CD68 and PR expression (p<0.05), without any other statistically significant correlations between IHC markers or between IHC and serological markers. Our study supports that endometriosis is an immune-dependent disease characterized by an abnormal morphological profile of T-cells and macrophages in endometriotic implants. Our study provides additional data useful in the understanding the immune milieu of endometriosis in the context of its complex pathogenic molecular mechanism. Further research is needed to develop new immunological therapeutic approaches, like immune checkpoint inhibitors administration or T-cell-targeted immunotherapy in these patients.

Symptomatic pericardial cysts and dilemmas in their diagnosis.

Pericardial cysts (PCs) or pleuropericardial cysts are rare congenital mediastinal lesions with an approximate incidence of one in 100 000 persons. Usually, they are asymptomatic, being incidentally discovered during a routine chest imaging examination or an autopsy exam. The study involved a retrospective evaluation of clinicopathological findings in a 6-year series of PCs, treated in the Clinic of Pulmonary Diseases, Iasi, Romania. A group of five cases of PCs, four females and one male, were evaluated. All patients displayed different symptoms, such as dyspnea, chest pain, chronic cough, fatigue, palpitation, and epigastric pain. The cystic lesions were located in the right and left cardiophrenic angle, in four cases, and in the central mediastinum in a single case. The lesions had a fluid content and a maximum diameter that ranged between 35 and 95 mm. The microscopic examination of the surgical resection tissues revealed a thin connective tissue wall without any associated smooth muscle cells. The loose connective tissue band was lined by a layer of mesothelial cells with no cellular atypia, which displayed discrete papillary projections, in one case. Although PCs are rare incidental findings, they should be considered in differential diagnoses of mediastinal cysts, especially as they are associated with non-specific symptoms. Furthermore, considering the possibility of development of severe complications, PCs should be thoroughly explored for suitable patients' management.

The clinicopathological challenges of symptomatic and incidental pulmonary hamartomas diagnosis.

Pulmonary hamartomas (PHs) are the most common benign lung tumors. Usually, they are asymptomatic and incidentally discovered during assessment for other diseases or during the autopsy exam. In this context, we have performed a retrospective analysis of surgical resections in a 5-year series of patients diagnosed with PHs in the Clinic of Pulmonary Diseases, Iasi, Romania, aiming to evaluate their clinicopathological features. A total of 27 patients with PH (40.74% males and 59.26% females) were evaluated. 33.33% of patients were asymptomatic, while the others exhibited variable symptoms, such as chronic cough, dyspnea, chest pain or weight loss. In most cases, PHs presented as solitary nodules, predominantly disposed in the right upper lobe (40.74% of cases), followed by the right lower lobe (33.34%), and left lower lobe (18.51%). The microscopic examination revealed a mixture of mature mesenchymal tissue, such as hyaline cartilage, adipose tissue, fibromyxoid tissue, and smooth muscle bundles, in variable proportions, associated with clefts of entrapped benign epithelium. A dominant adipose tissue component was observed in one case. PH was associated with a history of extrapulmonary cancer diagnosis, in one patient. Although considered benign lung tumors, PHs diagnosis and therapy may be challenging. Having in mind the possibility of recurrence or their occurrence as a part of specific syndromes, PHs should be thoroughly investigated for an appropriate patients' management. Their complex significance and the correlation with other types of lesions, including malignancies, may be further studied, by more extensive studies of surgical and necroptic cases.

Spongiotic reaction patterns in autoimmune bullous dermatoses (Review).

Spongiosis or a spongiotic reaction pattern is the histological hallmark of intercellular epidermal edema, viewed as clear spaces within the epidermis. Although considered a histopathological term, spongiosis has clinical correlations, with the variable degrees of spongiotic reaction leading to different dermatological findings. This review aimed to highlight the spongiotic reactive patterns found in different autoimmune bullous dermatoses, considering the paucity of publications in this domain. The pathogenesis of spongiosis assumes the passage of extravasated edema fluid from the dermis into the epidermis, frequently accompanied by dermal inflammatory cells, and classification of the spongiotic reaction patterns, as well as their associated spongiotic dermatitis, take into consideration the type and distribution of these inflammatory cells. It is mandatory to consider different reactive processes, specific for other skin disorders, which act as simulants of different spongiotic patterns for the diagnosis. Considering the possible transient occurrence, the heterogeneity and non-specificity of the histopathological features of these diseases, the diagnosis is very complex, requiring clinicopathological correlations and additional analyses. A deep insight into spongiosis pathogeny may open the perspectives of a classification refinement of autoimmune bullous dermatoses.

Thymolipoma - the frontier between hamartoma and neoplasia?

Thymolipoma is an uncommon benign thymus lesion, with a partially deciphered etiopathogeny, being most frequently diagnosed in young patients, regardless of gender. Incidentally diagnosed in asymptomatic patients, larger thymolipomas lead to symptoms related to neighboring mediastinal structures compression, with an intensity which is correlated with the mass size. Our review presents the main epidemiological, pathogenic, clinicopathological and morphological characteristics of this rare pathology. Sometimes, thymolipomas may be associated with paraneoplastic syndromes, which are alleviated by the mass complete surgical resection. Imagistics may orientate the diagnosis, which is certified by the microscopic examination of the resection specimens. Extensive thymectomy remains the current therapeutic option and new tools have been developed to increase the accuracy of the surgical procedure to avoid incidental lesions of the important elements of the anterior mediastinum. Although rare, thymolipomas should be considered in the differential diagnosis of mediastinal masses and of paraneoplastic syndromes.

Perivascular adipose tissue in cardiovascular diseases-an update.

The perivascular adipose tissue (PVAT) has been recently recognized as an important factor in vascular biology, with implications in the pathogenesis of cardiovascular diseases. The cell types and the precursor cells of PVAT appear to be different according to their location, with the component cell type including white, brown, and beige adipocytes. PVAT releases a panel of adipokines and cytokines that maintain vascular homeostasis, but it also has the ability of intervention in the pathogenesis of the atherosclerotic plaques development and in the vascular tone modulation. In this review, we summarize the current knowledge and discuss the role of PVAT as a major contributing factor in the pathogenesis of ischemic coronary disease, hypertension, obesity, and diabetes mellitus. The new perspective of PVAT as an endocrine organ, along with the recent knowledge of the mechanisms involved in dysfunctional PVAT intervention in local vascular homeostasis perturbations, nowadays represent a new area of research in cardiovascular pathology, aiming to discover new therapeutic methods.

The histopathological landscape of the major psoriasiform dermatoses.

Psoriasiform dermatoses represent a wide spectrum of inflammatory conditions, with several major forms represented by psoriasis, as the prototype of this category, followed by pustular psoriasis, Reiter's syndrome, pityriasis rubra pilaris, lichen simplex chronicus and large-plaques parapsoriasis. They create a diagnostic challenge, both clinical and histopathological, because of their complexity and frequent overlapping of the microscopical features. The characteristic histopathological features of psoriasiform reaction comprise extensive hyperkeratosis, with horizontally confluent but vertically intermittent parakeratosis, which alternate with orthokeratosis, thin granular layer, with relative frequent mitoses, uniform elongated and fused rete ridges, edematous superficial papillary dermis, with dilated capillaries, perivascular lymphocytic infiltrate, Munro's microabscesses, and spongiform pustules of Kogoj. Our paper aims to review the histopathology of major form of psoriasiform dermatoses and to emphasize the characteristic microscopical differences between them, for a better approach of the diagnosis as an important key for clinical and therapeutical management. Using the clinicopathological correlations, a thoroughly evaluation of the microscopical features and compartments distribution or special stainings and techniques, the range of differential diagnosis can be decreased and a more accurate diagnostic can be usually achieved. The insights into the pathogenic mechanisms can lead to new therapeutic opportunities targeted to the specific type of inflammatory lesion.

Ovarian hydatid cyst - systematic review of clinicopathological and immunohistochemical characteristics of an unusual entity.

Primary ovarian hydatid disease (HD) is a rare entity, produced by the larval stage of Echinococcus granulosus. HD commonly involves liver, lung, abdomen cavity, spleen and is unusually identified in pelvic organs. Based on our knowledge, the paper reviews 27 literature reports of ovarian HD, diagnosed during the last 20 years, providing a valuable database. Patients' ages ranged between 12-76 years, the gross appearance was that of 40-330 mm diameter hydatid cysts (HCs), 66.66% of them being primary. According to these reports, ovarian HD has non-specific clinical manifestations, such as abdominal or pelvic pain, nausea, dysmenorrhea or amenorrhea. The diagnosis may be achieved by abdominal ultrasound, computed tomography (CT) or magnetic resonance imaging (MRI), serological exams, such as eosinophilia (in 10-30% of patients) or indirect hemagglutination and immunoglobulin (IgG) antibodies detection. Ovarian HC microscopic pattern is characterized by three layers: pericyst or adventitia (host origin), germinal layer (endocyst), and laminated membrane (ectocyst). The immunoreaction triggered by parasites is initially rich in macrophages and neutrophils, followed by eosinophils and lymphocytes, with numerous cluster of differentiation 8 (CD8)-positive T-cells in active lesions and progressive forms. Concomitant ovarian diseases are relatively rare, being represented by borderline tumors (n=2 cases), mucinous cystadenoma (n=1 case), hemorrhagic cyst (n=1 case), and serous adenocarcinoma (n=1 case). In conclusion, the ovarian location of HD should be considered in any differential diagnosis of a cystic lesion, while it does not exclude synchronous ovarian tumors. These cases reinforce the necessity of better measures of prophylaxis and screening of HD in endemic areas.

The endometrial regeneration frontiers: from mechanisms to applications in regenerative medicine.

The endometrium is a unique and remarkable tissue characterized by a constant regeneration activity and this has been speculative for scientists, regarding its mechanism, regulatory factors, and their significance for fertility and endometrial pathology. Relatively recent scientific progresses due to genomics, proteomics, and transcriptomics have changed the knowledge in respect with endometrial regeneration and uterine-derived diseases. Our review is designed to highlight the recent progresses in understanding the endometrial physiology and its alterations involvement in uterine-derived diseases, addressing the current paradigm regarding endometrial regeneration, based on endometrial regenerative cells. In an attempt to explain the complex process of endometrial regeneration, different mechanisms have been proposed during time, from proliferation of basal glandular cells, to mesenchymal-epithelial transition, and lately to differentiation of stromal cells, based on endometrial regenerative cells or stem cells. Their unlimited potential of reconstruction of any type of tissue has been demonstrated and is currently in different trial stages in cell-based therapies of regenerative medicine, opening promising perspectives in severe or lethal diseases, by exploitation of stem cells. Currently, beside uterine acquired diseases and infertility, endometrial stem cells have been tested in a large spectrum of clinical applications. The great potential of endometrial cells for cell therapies arise from their accessibility, completely xeno-free derivation, allogenic use, possibility of large-scale therapeutic doses production, safety, reproducibility, and chance to overcome the drawbacks associated with autologous therapies. In order to overcome hostile environment of an injured tissue, the association of endometrial stem cells with other cells or added medium opens the perspective of specific combination available as standardized therapeutic means in the next future.

Adipocytes spectrum - From homeostasia to obesity and its associated pathology.

Firstly identified by anatomists, the fat tissue is nowadays an area of intense research due to increased global prevalence of obesity and its associated diseases. Histologically, there are four types of fat tissue cells which are currently recognized (white, brown, beige, and perivascular adipocytes). Therefore, in this study we are reviewing the most recent data regarding the origin, structure, and molecular mechanisms involved in the development of adipocytes. White adipocytes can store triglycerides as a consequence of lipogenesis, under the regulation of growth hormone or leptin and adiponectin, and release fatty acids resulted from lipolysis, under the regulation of the sympathetic nervous system, glucocorticoids, TNF-α, insulin, and natriuretic peptides. Brown adipocytes possess a mitochondrial transmembrane protein thermogenin or UCP1 which allows heat generation. Recently, thermogenic, UCP positive adipocytes have been identified in the subcutaneous white adipose tissue and have been named beige adipocytes. The nature of these cells is still controversial, as current theories are suggesting their origin either by transdifferentiation of white adipocytes, or by differentiation from an own precursor cell. Perivascular adipocytes surround most of the arteries, exhibiting a supportive role and being involved in the maintenance of intravascular temperature. Thoracic perivascular adipocytes resemble brown adipocytes, while abdominal ones are more similar to white adipocytes and, consequently, are involved in obesity-induced inflammatory reactions. The factors involved in the regulation of adipose stem cells differentiation may represent potential pathways to inhibit or to divert adipogenesis. Several molecules, such as pro-adipogenic factors (FGF21, BMP7, BMP8b, and Cox-2), cell surface proteins or receptors (Asc-1, PAT2, P2RX5), and hypothalamic receptors (MC4R) have been identified as the most promising targets for the development of future therapies. Further investigations are necessary to complete the knowledge about adipose tissue and the development of a new generation of therapeutic tools based on molecular targets.

Pointing at Ito cell, from structure to function (⃛ or Cinderella story in liver histology).

For more than 130 years, the hepatic stellate cells (also known as Ito cells) have been the object of numerous studies that aimed at their characterization both in normal and postlesional status, where they play a key role in fibrosis progression specific for chronic hepatic pathology. Converged studies on their histophysiology have revealed other functions, namely the involvement in liver embryological development and regeneration, metabolisms regulation, modulation of local inflammatory and immune reactions. Ito cells plasticity is surprising, as they are able to provide the connection between the complex sinusoidal and parenchymal microenvironment, influencing by autocrine and paracrine mechanisms the extracellular matrix content in tight correlation to growth and repair necessities. Last but not least, Ito cells take an active part in systemic homeostasis maintenance by their capacity to store and mobilize vitamin A, respectively. The evolving interest in their research will be undoubtedly followed by a better knowledge of the physiopathological sequences responsible for liver diseases, as new targets for the development of new therapeutic approaches directed toward improvement of prognosis and patients' quality of life.

Remodeling of basement membrane in patients with asthma.

The "bronchial remodeling" specific for the asthmatic disease consists in irreversible changes of the bronchial wall, including glandular and smooth muscle fibers hyperplasia and÷or hypertrophy, goblet cells hyperplasia, and thickening of basement membrane (BM). We aimed to analyze the BM thickness in asthma patients, in order to validate the relationship between its changes and the disease severity defined in agreement with the Global Initiative for Asthma (GINA) criteria. The study group has been formed of 38 patients with different degrees of severity of asthma established by spirometry using Forced Expiratory Volume in one second (FEV1), and two patients without asthma symptoms as controls. The specimens harvested by fibrobronchoscopy have been processed by paraffin embedding followed by Hematoxylin-Eosin (HE) and Periodic Acid-Schiff (PAS) staining. For each case, the BM measurement has been realized by a "point-by-point" method. Statistical analysis has been performed using SPSS 17 software, by applying non-parametric correlation tests. The quantitative assessment revealed a progressive increase in BM thickness during the course of the disease, from a mean value of 11.2 µm in stage 1 to that of 15.6 µm in stage 4. Even if this process has been noticed starting with the first stage of asthma, the differences in the BM size were statistically significant only for stages 1 and 3 (p=0.047), stages 1 and 4 (p=0.000), stages 2 and 3 (p=0.000), and stages 3 and 4 (p=0.000). Spearman's test has shown an opposite correlation between the BM thickness and asthma severity defined by FEV1 values (r=-0.86, p<<0.01, 95% CI). Our study demonstrates that the collagen deposition at the epithelium-connective interface is initiated in early stages of asthma and continues in a progressive modality, the BM thickening being correlated with the disease severity. Thus, we support the concept of biological consequences of BM thickening in asthma pathogenesis, a mechanism still incompletely deciphered.

Macrophage profile in primary versus secondary liver tumors.

Macrophages are important components of the tumor-associated infiltrate and are qualified as one of the major players of the cancer-related inflammation. It was shown that tumor cells can either stimulate or mediate apoptosis of tumor-associated macrophages (TAMs). To date, there is no general agreement regarding the influence of TAMs and their numbers on the progression of hepatocellular carcinoma (HCC) and hepatic metastases (HM). To analyze the presence of TAMs and compare their numbers in intratumoral (IT) and peritumoral (PT) areas with the clinical outcome of HCC and HM patients. Biopsies from 35 HCC and 39 HM cases were analyzed. Clinical and follow-up data was enrolled for each case; the colorectal cancer was the origin of 26 HM patients. TAMs were identified by immunohistochemistry using anti-CD68 monoclonal antibody. The quantitative assessment was performed by determining the mean number of CD68-positive cells in IT and PT areas in HCC and HM. Two threshold methods were applied: threshold 1 (T1) was calculated with the use of (-log) Cox method; threshold 2 (T2) was considered as 1/3 TAMs number of group's mean. For statistical analyses Mann-Whitney U-test, Spearman's correlation, Cox proportional hazard and Kaplan-Meier tests were applied. To date, 36.12% HCC and 27.78% HM patients were alive, median survival was 5 and 17 months for HCC and HM, respectively (P = 0.05). We found significant two-fold decrease of TAMs numbers between IT vs. PT territories in both HCC and HM. A positive correlation between numbers of PT and IT TAMs was observed in HM group (rs = 0.48, P < 0.05) but not in HCC. The number of TAMs was not associated with any studied clinical factor. Univariate Cox regression analysis showed that tumor stage ≤ II (P = 0.01) and increased number of PT TAMs (P = 0.06, only when T2 value was applied) were associated with favorable prognosis in HCC (HR = 2.614 and 2.457, respectively). Univariate and multivariate Cox analyses in HM revealed favorable prognosis for histological grade ≤ G2 and one lobe tumors (P = 0.021 and 0.045; HR = 0.395 and 0.438, respectively). Survival analysis retained the impact of increased TAMs numbers in peritumoral areas (P = 0.03), tumor stages in HCC (P = 0.007), lobes' number (P = 0.007) and histological grade (P = 0.005) on HM patients' outcome. In HCC and HM the low number of TAMs in intratumoral areas was related to the tumor cell microenvironment. The increased peritumoral TAMs number in primary liver tumors was associated with better prognosis.

A morphometric approach in breast cytology--geometrical descriptors in the differentiation between benign and malignant lesions.

This study focuses on the analysis of geometric descriptors that can be applied in breast cytology, and their correlation with the qualitative features, with the aim to underline the differences between the benign and malignant cell profile. The morphometric investigation was performed on smears obtained by fine needle aspiration, 10 cases (group 1) diagnosed as benign and 10 cases (group 2) as malignant. For group 2, the malignancy was histopathologically confirmed on the surgical resection specimen. The sequence of automated operation, previously reported by us, permitted the extraction of the following geometrical descriptors: cytoplasmic area, nuclear area, nucleo-cytoplasmic ratio, equivalent diameter and form factor. We analyzed the differences between the benign and malignant morphometric features, and the correlation between the malignant morphometric features and cytological, respectively histological grading. Statistically significant difference in cytoplasmic areas, nuclear areas, value of nucleo-cytoplasmic ratio and equivalent diameter was noted between group I and II. For the form factor, we did not register statistically significant differences. For group 2, the correlation between the morphometric features and cytological grading revealed that the nuclear area is the most valuable descriptor, due to the significant differences between the three successive grades of cytological severity, followed by the cytoplasmic area and equivalent diameter, their numerical values presenting significant differences between cytological grade 1 and 3, and 2 and 3, respectively. The statistical analysis between the morphometric features and histological grading showed that nuclear area and equivalent diameter are the most viable indicators, due to the significant differences present between the three successive grades of pathologic severity, followed by cytoplasmic area (significant differences only for grade 2 versus 3) and for nucleo-cytoplasmic ratio (significant differences only for grade 1 versus 2). The form factor does not provide information that could be correlated with the cytological or histological grading. The defined morphometric features enable the characterization of benign and malignant cells and provide objective criteria that could support a differentiation of benign from the malignant pathology in the cytological diagnosis.

High variability in MMP2/TIMP2 and MMP9/TIMP1 expression in secondary liver tumors.

The assessment of MMPs/TIMPs expression in various primary tumors has potential prognostic values. Considering the paucity of studies in secondary liver tumors, our aim was to study the expression of MMP2, MMP9, TIMP1, and TIMP2 in secondary hepatic cancer, focusing on their variability in the malignant cells. The study group included 25 cases of liver metastases of colorectal cancer, diagnosed and surgically treated at "Sf. Spiridon" University Hospital of Iassy, Romania. Immunohistochemistry for MMP2, MMP9, TIMP1, and TIMP2 has been performed, followed by the semi-quantitative assessment of the markers using a scoring system based on the positive tumoral cells percentage and the staining intensity. The expression of investigated markers revealed an increased staining variability. The scores showed that MMP2/TIMP2 and MMP9/TIMP1 immunoreactivity was extremely heterogeneous within the analyzed group, with a dominant weak expression for MMP2 and MMP9, in contrast to strong TIMP2 and TIMP1 expression. Ten different patterns of expression of the investigated markers have been identified. No major differences between the expression of MMP2 (14 positive cases) and MMP9 (16 positive cases) could be detected. Our results sustain the inverse correlation between MMP and the correspondent TIMP expression, supporting the hypothesis that MMPs/TIMPs balance has mainly an inhibitory effect on invasiveness. Our study demonstrated that tumoral cells are adapted to MMPs:TIMPs production in the liver microenvironment. The lack of significant differences between MMP2 and MMP9 expression shows that the activity of both MMPs is independent, without reciprocal influences.

CLDN3 expression and significance - breast carcinoma versus ovarian carcinoma.

Claudins (CLDNs) are transmembrane proteins, as normal constituents of the architecture of tight junctions. Recent studies support their involvement in carcinogenesis, as changes in CLDNs structure result in alterations in tight junctions' structure and function, facilitating malignant transformation. We aimed CLDN3 investigation in both breast and ovarian carcinoma, targeting the identification of its expression differences. The immunohistochemical assessment was performed on 20 cases of breast carcinomas (Group 1) and 19 cases of epithelial ovarian carcinomas (Group 2). Firstly, the specific panel for the molecular classification was applied for specimens of the first group. Then, all the specimens were immunostained for CLDN3 and a semi-quantitative evaluation was made, based on the percentage of positive cells and the intensity of staining. In Group 1, in the ER positive category, CLDN3 was overexpressed in five cases (four cases of luminal A and one case of luminal B subtype, respectively), negative in three cases (luminal A subtype) and weakly expressed in a single case (luminal A subtype); in ER negative category, CLDN3 expression was strong in four cases (one case of Her2/neu subtype and three cases of basal-like subtype), negative in two cases (normal breast-like subtype) and weak in five cases (one case of Her2/neu subtype, one triple-negative subtype, and three basal-like subtype). In Group 2, CLDN3 was overexpressed in 15 cases, histopathologically diagnosed as serous (10 cases), mucinous (two cases), endometrioid (two cases), and mixed carcinomas (one case); a weak expression was noticed in a single case, of the serous subtype; CLDN3 was undetectable in three cases (one serous, one clear cell, and one endometrioid type). Our comparative analysis of CLDN3 profile in breast and ovarian cancer clearly indicates organ specificity.

Relationship between Kuppfer cells, inflammation, and fibrosis in chronic hepatitis B and C.

Kupffer cells are liver parenchymal components of the immune system able of intervention in pathogenic mechanisms involved in viral hepatitis and tumoral lesions. Our study aimed to evaluate the Kupfferian hyperplasia in chronic hepatitis B and C, and to correlate with the severity of liver lesions, focusing on the modality in which Kupffer cells modulate the necroinflammatory events and fibrosis specific for chronic hepatitis morphologic substrate. We investigated 33 cases with chronic hepatitis B and 38 cases with chronic hepatitis C, diagnosed according to Ishak score (in chronic hepatitis B and C) and METAVIR score (in chronic hepatitis C). Kuppfer cells were immunohistochemical labeled, by using an anti-CD68 antibody and heat-induced epitope retrieval (HIER) technique. The sinusoidal reaction expressed by CD68 (+) cells hyperplasia progressively increases along with the intensity of necroinflammatory activity and with the amplitude of fibrosis lesions. Statistically significant differences between Kupffer cells number and the degree of necroinflammatory activitiy and fibrosis, respectively (Ishak score) have been identified in both types of chronic hepatitis. However, no significant differences have been registered when comparing the Kupffer cells number corresponding to each degree of necroinflammatory activity and of fibrosis, in chronic hepatitis B versus C, respectively. Our results demonstrate the relationship between Kupffer cells and the severity degree of the disease, without differences between chronic hepatitis B and C. Consequently, we may appreciate that the chronic hepatitis C specific lesions progression, different from that of chronic hepatitis B, is influenced by a different behavior of Kupffer cells, and not by their effective

Relationship between immunohistochemical assessment of bronchial mucosa microvascularization and clinical stage in asthma.

Although hardly ever used in current practice, fibrobronchoscopy may provide interesting histopathological-clinical correlations in patients diagnosed with different stages of evolutive asthma. The aim of the study was to evaluate the correlation between semi-quantitative microvascularization features and the asthma severity assessed according to the GINA classification 2006. Our study group consisted in 21 patients diagnosed with asthma of different stages of severity and two-control patients investigated by fibrobronchoscopy with associated biopsy. The tissue fragments underwent standard processing procedures for the immunohistochemical exam, using CD34 as microvascularization marker. The semi-quantitative analysis was based on the "hot spot" method and on a score system that corresponds to the microvessels density. The statistical analysis of the correspondence between CD34 score and clinical parameters was performed using the SPSS 17 software, applying non-parametric correlation tests. The CD34 evaluation showed an increase in blood vessels count in all asthmatic patients in comparison to the control group and a close correlation with the asthma severity, reflected by the FEV1 values. The statistical analysis showed an inverse correlation between FEV1 [%] values and CD34 expression (r=-0.93, p<<0.01). Our data concur to other research reports, supporting the hypothesis that angiogenesis initially facilitates the edema development and later on appears to be involved in the bronchial wall thickening, as a component of the chronic inflammatory response, with concomitant distensibility reduction. The bronchial mucosa microvascularization evaluation opens new perspectives for advanced therapies, with beneficial effects for asthmatic patients' life quality.

Operational scores in the diagnosis of chronic hepatitis. A semi-quantitative assessment.

Starting from the quantification of the specific lesions for chronic hepatitis B and C, our study focused on (i) the correspondence between the necroinflammatory activity and the fibrosis stage ascertained through the Ishak scoring system, (ii) the classification overlaps and differences of Ishak vs. METAVIR score. The study group consisted of 202 cases with chronic hepatitis B and 751 cases with chronic hepatitis C, diagnosed based on liver biopsies. The fragments of hepatic tissue were routinely processed and stained with Hematoxylin-Eosin, trichrome Szekely, Gordon-Sweet silver impregnation, and Periodic Acid-Schiff. A semiquantitative evaluation was performed using the Ishak (for hepatitis B and C) and the METAVIR (for hepatitis C) scoring systems. Our results revealed that the comparison between hepatitis B and C, based on the necroinflammatory activity and fibrosis, is able to offer through the numeric values of the Ishak scoring system accurate proofs, which support the aggressivity of hepatitis C, because it develops fibrosis more quickly, even on the background of mild necroinflammatory activity. Also, our data showed that the necroinflammatory activity and the fibrosis are not processes which progress in a consistent pattern. The application of the METAVIR scoring system for the cases with chronic hepatitis C confirmed that there is not a direct correlation between necroinflammation and fibrosis. The Ishak scoring system provides through the wide range of numeric values attributed for the evaluation of necroinflammatory activity and fibrosis far more precise criteria for the appraisal of the degree of damage to the hepatic parenchyma at the time of the diagnosis. Supplementary, the METAVIR scoring system allows for the hepatitis C an assessment of the entire histologic activity, including the interface hepatitis and the associated lobular necrosis components. The scoring systems have unavoidably strengths and weaknesses, but the choice of a specific one must reflect the consensus between the pathologists and the clinicians, relying on their experience.

Immunohistochemical assessment of p16, COX-2 and EGFR in HPV-positive cervical squamous intraepithelial lesions.

The protein capsid L1 of the human papilloma virus (HPV) - a key factor in the cervical carcinogenesis - is considered, together with p16, EGFR and COX-2, a characteristic marker for the evaluation of the malignancy progression and prognostic, in terms of tumoral aggressiveness. The purpose of the present study was to make a comparative assessment between the immunohistochemical pattern of p16, EGFR and COX-2 and immunochemical expression of L1 HPV capsid protein, in low grade and high-grade cervical squamous intraepithelial lesions, in order to determine the relationship of these tumoral markers with the infection status of HPV, and their practical applicability in patients diagnosis and follow-up. The study group included 50 women with cytological and histopathological confirmed LSIL (low grade SIL) and HSIL (high-grade SIL). The immunoexpression of L1 HPV protein was assessed on conventional cervico-vaginal smears and EGFR, COX-2 and p16 were immunohistochemically evaluated on the corresponding cervical biopsies. From all cervical smears, the HPV L1 capsid protein was expressed in 52% of LSIL and 23% of HSIL. From all cervical biopsies, p16 was positive in 64% of LSIL, 82% of CIN2 and 100% of CIN3, EGFR was overexpressed in 67% of HSIL (56% CIN2 and 43% CIN3) and 32% LSIL. For COX-2, the Allred score was higher in HSIL when compared to LSIL. Our data revealed 33 cases belonging to both LSIL and HSIL categories with the same Allred score. Immunochemical detection of L1 capsid protein, on cervico-vaginal smears, indicates an immune status induced by the HPV infection and may offer prognosis information, mainly in LSIL lesions. The assessment of p16, EGFR, and COX-2 allows to an integrative approach for the progression of squamous intraepithelial lesion, associated or not with the HPV infection.