Correlations Between Intratumoral Interstitial Fibrillary Network and Vascular Network in Gleason Patterns of Prostate Adenocarcinoma.

PURPOSE: The study aims to assess the correlation between stromal fibrillary component (SFC) and vascular density (VD) in Gleason architectural patterns of prostate carcinoma. MATERIALS AND METHODS: 680 digital images of prostate adenocarcinoma labeled following both Gleason and Srigley systems were acquired with X20 objective from serial sections, one stained using Gömöri technique for SFC and one immunomarked with anti-CD34 antibody for vessels. The SFC amount and VD were determined and compared. Gleason patterns were divided in: "Solid" group (Gleason 3a, 3b, 4b, 5b) and "Necrotizing" group (Gleason 3c 4a and 5a). For each parameter were assessed: the lowest value (VMIN), the highest value (VMAX), the half range value (HRV), mean value (AV), standard deviation (STDEV), mean value + standard deviation (AV+ STDEV ) and mean value + standard deviation (AV+ STDEV). The Pearson product-moment correlation coefficient and the χ2 test were used. RESULTS: The relationship between SFC and VD values had an inverse, descending correlation in Gleason 2 pattern and a direct, ascending correlation in Gleason 4 and 5 patterns. In Gleason 3 pattern, although the trend line had a direct ascending trend, it was not validated by the Pearson's and χ2 tests. However, SFC and VD values had a direct, ascending correlation for all determinations (p<0.05), but also for "Solid" (p<0.05) and "Necrotizing" (p<0.05) groups. CONCLUSIONS: The assessment of the relationship between the two main components of the intratumoral stroma in prostate carcinoma showed that they are evolving in a parallel manner. There is still need for studies on larger groups in order to decipher and more clearly define the way the stromal microenvironment is remodeling according to the malignant cell population degree of differentiation.

Correlations between intratumoral vascular network and tumoral architecture in prostatic adenocarcinoma.

The authors made a preliminary assessment of possible correlations between the intratumoral vascular density (IVD) and the architectural tumoral patterns described by Gleason. The studied material consisted of samples obtained by transurethral resection from 34 patients diagnosed with prostatic adenocarcinoma. Ten fields, five for dominant and five for secondary identified patterns of each case, with no necrosis were selected randomly from CD34 immunomarked sections using ×20 objective. IVD increased with Gleason pattern both for the entire group, but also for "solid" phenotype group of subtypes up to pattern 4, respectively subtype 4B. In "necrotizing" phenotype group of subtypes, IVD had a decreasing trend from the better-differentiated subtypes to the poorest one. These preliminary data showed that the intratumoral vascular network reacts differently to the loss of tumoral differentiation in the two groups of Gleason subtypes suggesting the existence of two different populations of malignant cells.

Endometriosis - clinical approach based on histological findings.

Endometriosis is a benign disease defined by the presence of endometrial glands and stroma outside of the uterus and is associated with both pelvic pain and infertility. The most common sites of endometriosis, in decreasing order of frequency, are the ovaries, anterior and posterior cul-de-sac, posterior broad ligaments, uterosacral ligaments, uterus, fallopian tubes, sigmoid colon, appendix, and round ligaments. The main treatment is surgical, but often-microscopic islands of endometrial tissue remain, which proliferate and are responsible for relapses. We tested the efficacy of two drugs (Medroxyprogesterone and Triptorelinum), administered for six months to prevent recurrence after surgery. Treatment with Medroxyprogesterone was 100% effective in terms of relapse, while Triptorelinum could not prevent recurrence of endometriosis.

Role of innate immune receptors TLR2 and TLR4 as mediators of the inflammatory reaction in human visceral adipose tissue.

White adipose tissue from different locations is characterized by significant differences in the structure of adipocyte "secretoma". Fat accumulation in the central-visceral depots is usually associated with a chronic inflammatory state, which is complicated by the metabolic syndrome. Recently, the adipose tissue was emerged to have an essential role in the innate immunity, adipocytes being considered effector cells due to the presence of the Toll-like receptors (TLRs). In this study, we compared the expression of TNF-α, TLR2 and TLR4 in peripheral-subcutaneous and central-peritoneal adipose depots in three different conditions - lean, obese and obese diabetic - using immunohistochemistry. Our results suggest a correlation between the incidence of the stromal vascular cells and adipocytes TNF-α and TLR4 in the visceral depots in strong correlation with adipose tissue expansion. TLR2 positive cells were seen in the peripheral depots from all groups without any association with fat accumulation. These results focus on the existence of a new pathogenic pathway, the activation of TLR4, for the involvement of visceral adipose tissue in the activation and maintenance of the inflammatory cascade in obesity.

Morphometric findings in avascular necrosis of the femoral head.

Avascular necrosis of the femoral head is an illness with a controversial etiology, the trigger event being the suppression of blood flow to the femoral head. The disease affects mostly young adults within their third and fifth decade, the majority of the patients being men. The main risk factors are trauma, chronic alcohol consumption, smoking, corticotherapy. The main goal of our study is to describe the morphometric changes found in the bone tissue of patients diagnosed with avascular necrosis of the femoral head, with different risk factors, by comparing the area of bone trabeculae inside the area of necrosis with that from the adjacent viable tissue. The morphometric study used biological material from 16 patients with ages between 29 and 57 years, who underwent surgery for avascular necrosis of the femoral head. They were admitted in the Orthopedics Department at the Emergency County Hospital in Craiova between 2010 and 2011 and were split into four groups. Group I presented trauma as the main risk factor, Group II had corticotherapy as the defining risk factor, Group III presented chronic alcohol consumption and Group IV was represented by the patients who smoked and exhibited chronic alcohol consumption. There was not a significant statistical difference between the areas of bone trabeculae of the four groups when we compared viable bone tissue to the necrotized one. Knowing the risk factors of the avascular necrosis of the femoral head is critical to the management of the disease, because diagnosing it in an early stage is a necessity for obtaining a good result for conservative treatment.

Histological and immunohistochemical changes of the myocardium in dilated cardiomyopathy.

Dilated cardiomyopathy is a major cause of heart failure and a major cause of morbidity and mortality. It is a multifactorial disease that includes both hereditary and acquired forms. It is estimated that around 20-35% of patients with dilated cardiomyopathy have hereditary forms. It is the third most common cause of heart failure and the most common cause of heart transplant. Dilated cardiomyopathy can be a secondary condition of many diseases such as coronary heart disease, diabetes, pheochromocytoma, infections, malnutrition, ingestion of toxic substances (alcohol, cocaine), ingestion of chemotherapeutic drugs, autoimmune diseases. In our study, we aimed to describe the changes of myocardial cells and interstitial connective tissue in patients clinically diagnosed with alcoholic dilated cardiomyopathy. The material studied consisted of heart fragments sampled from the left ventricle (LV) during necropsy from a total of 28 patients, aged between 58 and 73 years, with a clinical and laboratory diagnosis of dilated cardiomyopathy, hospitalized in the Cardiology Center of the Emergency County Hospital of Craiova in 2009 and 2010. In dilated cardiomyopathy, myocardial muscle fibers appeared slightly elongated or wavy, with hypochromatic, heterogeneous, vacuolar sarcoplasm, by a decrease of myofibril numbers. Lipofuscin granules were frequently seen in the sarcoplasm. Nuclear changes were consistent with sarcoplasmic alterations. Changes of the interstitial connective tissue were sometimes extensive and sometimes barely noticeable. The most common alteration of this structure was the onset and development of a mainly perivascular collagen fibrillogenetic process.

Chronic gastritis with intestinal metaplasia: clinico-statistical, histological and immunohistochemical study.

Chronic gastritis has a high incidence in adults, causing progressive destruction of glandular structures, favoring the development of gastric atrophy. The association of chronic gastritis with intestinal type metaplasia of gastric mucosa has a poor outcome as intestinal metaplasia is regarded as a precancerous lesion. Metaplasia is common in patients with Helicobacter pylori infection and also heavy smokers. The aim of our study was to evaluate the relationship between chronic gastritis and intestinal metaplasia. The study was conducted on a total of 1218 patients, aged between 5 and 90 years, who presented for dyspeptic disorders in the period 2007-2010 and were examined clinically and endoscopically. During the gastroscopic examination, fragments of gastric mucosa were collected for the histopathological study and for highlighting the H. pylori infection. For the histopathological study, the Hematoxylin-Eosin and PAS-Alcian Blue stains were performed, while for the immunohistochemical study the anti-TAG72 and anti-PCNA antibodies were used. A diagnosis of gastritis was established in 615 patients, representing approximately 50.5% of all cases. Most cases with gastritis were found in people of middle age. Gastritis was present in almost all age groups, from teenagers to the elders. Of the 615 cases of gastritis, urease test was positive in 353 patients, representing approximately 57.40% of all patients with gastritis. Histopathological examination identified the presence of intestinal metaplasia in 61.60% of patients with chronic gastritis, mostly complete metaplasia. PCNA immunohistochemistry revealed that cell proliferation processes are intensified in intestinal metaplasia. This study highlights the importance of chronic gastritis, intestinal metaplasia, and H. pylori infection in the etiopathogeny of gastric cancer.

Clinical and morphological correlations in acute ischemic stroke.

We studied the clinical and histopathological changes in twenty-seven cases of acute ischemic stroke, aged between 65 and 75 years. All deaths occurred within 30 days after stroke. The aim of our study was to establish the clinical and histological correlations in acute ischemic stroke to detect prognostic factors. Brain lesions after acute stroke were observed in all regions. Our study describes the heterogeneity of brain injury after acute ischemic stroke with the participation of all brain components and the chronology in which these lesions develop and evolve. By histological and immunohistochemical studies, we identified neuronal, glial and vascular damage. The neurons had undergone in the area of lesion a process of necrosis, ballooning or condensation process. In the ischemic penumbra, we observed the presence of red neurons. Vascular lesions were represented by the discontinuity of capillaries, always associated with a marked perivascular edema. The following clinical and morphological correlations were established: liquefactive necrosis, astrocyte gliosis, phagocytosis phenomena are the more intense the later the death of the patient; apoptosis phenomena are the more intense the faster the death of the patient; the entire cerebral microcirculation presented microscopic modifications following the ischemic strokes, regardless of the time since the lesion occurred and the histological examination was made; the major neurological complications of the ischemic stroke - the hemorrhagic transformation phenomena, cerebral edema, were microscopically objectified, regardless of the time since the lesion occurred and the histological examination was made.

Correlations between intratumoral interstitial fibrillary network and tumoral architecture in prostatic adenocarcinoma.

The authors made a preliminary assessment of possible correlations between the amount of intratumoral stromal fibrillary components (ISFC) and the architectural tumoral patterns described by Gleason. The studied material consisted of samples obtained by transurethral resection from 34 patients diagnosed with prostatic adenocarcinoma. Ten fields, five for dominant and five for secondary identified patterns of each case, with no necrosis were selected randomly from Gömöri stained sections using ×20 objective. ISFC-ratio increased with Gleason pattern both for the entire group but also for "Necrotizing" phenotype patterns and "Solid" phenotype patterns, excepting the subtype "4A" where the stromal compartment was reduced by the expansion of tumoral ducts enlarged by growing tumoral intraductal cribriform masses. These preliminary data showed that stromal microenvironment try to adapt to the loss of tumoral differentiation by increasing the amount of fibrillary components of intratumoral stromal compartment.

Histopathological and immunohistochemical aspects in chronic suppurative maxillary rhinosinusitis.

Chronic rhinosinusitis is a multifactorial disease with pathophysiological mechanisms, which remain unclear, and with a high prevalence worldwide. They generate social problems due to the high number of days of leave and relatively elevated medical expenses. The histopathological and immunohistochemical study that we conducted revealed many lesional aspects of the epithelium of the sinus mucosa, which ranged from hypertrophy, hyperplasia and metaplasia, to erosion and discontinuities. In the chorion of the sinus mucosa there was an inflammatory infiltrate composed mainly of lymphocytes, plasma cells and macrophages, and also a highly developed vascular network. Among immune cells, T-cells appeared to be more numerous than B-lymphocytes and macrophages. We believe that microscopic changes are due mainly to microscopic organisms that make up the biofilm of the sinus cavity, whose virulence has been more or less influenced by exogenous or endogenous factors.

Study of cellular changes induced by moderate cerebral ischemia achieved through internal carotid artery ligation.

Reduced cerebral blood flow beyond the compensatory mechanisms leads to cerebral hypoxia. Hypoxia causes various lesions of neurons, glial cells and cerebral blood vessels, depending on its duration and intensity. In our study, we reduced cerebral blood flow in the experience animal on average by 30%, by right internal carotid artery ligation. Fifteen days after the onset of hypoxia, by histology and immunohistochemical studies, we identified neuronal, glial and vascular damage. Lesions of nerve and glial cells ranged from changes of cytoplasmic tinting with the development of "red neurons", to neuronal and glial cytolysis with areas of focal necrosis. Vascular lesions were represented by the collapse, fragmentation and discontinuity of capillaries, always associated with a marked perivascular edema.

Pericytes and myofibroblasts reaction in experimental thermal third degree skin burns.

Pericytes and myofibroblasts are two types of connective cells that appear in large quantities in reparative processes. In order to evaluate their response and any relationships between them, we have inflicted third degree skin burns to seven groups of five common Wistar rats each from which we sampled granulation tissue at three days interval for histological and immunohistochemical studies. Alpha-SMA and CD34 antibodies revealed that granulation tissue myofibroblasts do not express CD34 and do not arise from pericytes. The parallel arrangement of myofibroblasts in relation to the burned wound surface suggests that their main function is mechanical traction on granulation tissue and wound edges.

The importance of tumor proliferation markers in assessing lesions of the palatine tonsil.

Oropharyngeal cancer, and especially squamous cell carcinoma, is one of the most common cancers worldwide, and its incidence is increasing, with the palatine tonsil being one of the main locations. The etiopathogenic factors, together with its location as well as the available immunohistochemical methods, make this type of cancer an accessible one in terms of diagnosis. However, it is usually diagnosed in late stages. Therefore, we tried to elucidate the causes of treatment failures and development of local recurrence. For this, we reassessed the proliferative pattern of tonsil lesions using the anti-p53, anti-PCNA and anti-Ki67 antibodies on 73 tonsil fragments collected after curative surgery on adults aged between 28 and 86 years. Following the reevaluation of the histopathological examination using markers for cell proliferation, the diagnosis was modified in 16 cases, representing about 22% of the cases take into study. By using immunohistochemical markers in the histopathological examination the diagnosis is improved, leading to a more appropriate therapeutical approach.

Angiogenesis assessment in experimental third degree skin burns: a histological and immunohistochemical study.

In the past 30 years, after the discovery of vascular proliferation factors, angiogenesis is one of the most intensively studied fields. Restoring the vascular network after burn injury is essential for healing, as it brings oxygen and nutrients to injured tissues, improves the contribution of inflammatory cells and prepares the damaged area for repair and tissue regeneration. To study the process of angiogenesis we used seven groups of five animals, each of adult Wistar rats, which were inflicted with third degree skin burns. From each group of animals, we sampled at successive intervals of three days the entire burnt wound with a ring of surrounding normal skin. Sampled skin fragments were processed for paraffin inclusion, sectioned with a microtome and stained with Hematoxylin-Eosin or Masson trichrome. The samples were also analyzed using single chromogenic immunohistochemistry or double immunofluorescence for the presence of CD34 and alpha smooth muscle actin (α-SMA). Angiogenesis process started at about three days after the burn infliction, with the appearance of tubular structures lined by CD34-positive cells. Subsequently, these cells showed intense proliferative activity that generated a network that included progressive neovascularization around the wound surface. Maximum vascular proliferation occurred at 9-15 days after injury, when the number of capillaries reached 229/mm², and the total area of capillary angiogenesis at 100.27 µm² (about 10% of the section area). Subsequently, the process of angiogenesis was gradually reduced, but remained at moderate levels after wound healing. During the process of angiogenesis, there was a very close relationship between CD34-positive cells and pericytes (as α-SMA-positive).

Histological and ultrastructural analysis of the seminiferous tubule wall in ageing testis.

The authors evaluated possible morphological changes of basement membrane (BM) and lamina propria (LP) of seminiferous tubule wall (ST) related to ageing. Surgical samples of testicular tissue from 28 cases with orchiectomy for prostate adenocarcinoma were processed for light microscopy and transmission electron microscopy (TEM) examination. Seven age groups (AgGr) between 50 and 80 years were designed. Tissue samples were immunomarked for collagen IV and smooth muscle actin. Images were acquired and measured with a specialized software. Thirty ST were randomly selected, with ×40-objective, for each case. Five random determinations for each ST and each parameter were performed. Mean values/tubule, case and AgGr were calculated for each parameter. Regression line (RL), slope and significance test for slope were determined for each parameter correlation with ageing. BM mean value was around 0.5 µm, with narrow limits of ranging in AgGr but more extended individual limits. RL showed discrete decreasing trend with ageing but without an obvious statistical correlation. LP mean value was around 6 µm, also with narrow limits of ranging in AgGr and more extended individual limits. RL decreased discretely with ageing but without an obvious statistical correlation. TEM showed more prominent BM material and more collagen fibers and less fibroblasts in LP of older AgGr and higher fibroblasts density in LP of younger AgGr. Our results showed that BM thickness is apparently decreasing with ageing whereas LP presents extremely variable degenerative changes, with a "mosaic", focal distribution and no tendency to advance with ageing.

Preliminary study of bipolar hip prosthesis - influence of acetabular bone interactions on bone morphology.

Periprosthetic bone changes following hip arthroplasty are yet to be completely described. The material consisted of imagistic records (X-ray films, CT and MRI scans) and of acetabular bone tissue sampled from 14 cases with femoral head prosthesis and revision of the prosthesis fixed and decalcified in Duboscq-Brazil solution and stained with Hematoxylin and Eosin, trichrome van Gieson and trichrome Masson. Acetabular bone is home of a great variety of morphological changes that can be divided in degenerative and regenerative changes seen in both compact and trabecular components but only inside the maximal pressure area of the acetabular roof. Our preliminary morphological study revealed the existence of an adaptation effort to the mechanical stress materialized through a dynamic process of bone remodeling in the maximal pressure area.

Correlations between intralobular interstitial morphological changes and epithelial changes in ageing testis.

The study focuses on the possible influences of intra (I) lobular (L) stromal compounds [intertubular spaces and seminiferous (S) tubule (T) wall (W)] morphologic changes on S epithelium (E) during ageing process. The material consisted of surgical samples of testicular tissue from 192 patients with orchidectomy for prostate carcinoma. Seven age groups were designed, from 50 to 80 years. Tissue samples were fixed in neutral buffered formalin, embedded in paraffin stained with HE, Goldner and Gömöri and immunomarked (in a subgroup of 28 cases) for smooth muscle actin, collagen IV, and CD34. SE had an uneven involution, both individually and inter-individually, but with normal spermatogenesis in many of ST. E degenerative changes were seen mainly in L periphery. Different stages of maturation arresting were more frequent in older patients. IL septae had changes with extremely variable intensity, dispersed mainly in L periphery, without significant spread and without extensive trend with ageing. Leydig cells showed focal hyperplasia without extensive trend related with ageing. STW presented strictly in the internal layer of lamina propria (apposed to basement membrane of ES) a focal sclerosis, with variable extension concerning its presence, thickness and T circumference (T without sclerosis, with focal sclerosis and with fibro-hyaline "collar" - FHyC) but not related with ageing. IL arteriolae showed focal areas of degeneration with a wide individual and inter-individual range of intensity and extension, but not related with age. Capillary network (CN), with both its peri-T and intramural segments, was present in all age groups, with no quantitative endothelial changes and decreasing only in very old cases. FHyC was often associated with E atrophy. STW focal sclerosis could explain focal degeneration of SE in senescence, although CN undergoes no significant changes.

Study of vascular microdensity in areas of cerebral ischemia on experimental model.

Stroke is the third leading cause of death and the first cause of disability with an increasing incidence, especially of the ischemic type. There is no effective curative treatment for stroke and therefore the therapy for this disease currently relies on identifying patients at risk and instituting preventive measures. Since ischemic stroke in middle cerebral artery territory (and with preferential localization in the striated nuclei) is the main type of stroke in humans, animal models obtained by surgical ligation of the middle cerebral artery (MCAO) are valuable tools for the fundamental study of this disease. In this study, we investigated the morphological and immunohistochemical remodeling of the tissue after stroke and in particular the vascular component in a MCAO murine model. The analysis included the sequential sacrificing of animals at 1, 14, 28 and 60 days post-stroke and brain processing for paraffin embedding and sectioning. Our results show a gradual revascularization of the lesion as we move away from the time of the surgical intervention. This effect is accompanied by the development of an increasingly dense glial scar at the periphery of the lesion. The perilesional area itself, the penumbra, is characterized by minimal histological changes (such as eosinophilic neurons), but also by an increased expression of activated caspase 3 as a sign of apoptotic neurons and glial cells. Our study confirms the potential of the organism in its attempt to revascularize the injured area, and raises questions on the role of the glial scar in limiting the process of neovascularization.

Morphological experimental study of bone stress at the interface acetabular bone/prosthetic cup in the bipolar hip prosthesis.

By calculating the tension and distortion of the elements composing the bipolar prosthesis under extreme conditions encountered in real life using a special post-processing program, we established the variation curves of the contact pressure at the hip bone-cup, armor-cup and cup-femoral head interface. By comparing the data obtained from all the examined cases, important conclusions were drawn regarding the influence of tension and pressure distribution on the structural integrity and biomechanics of the prosthesis, as well as the acetabular wear and tear, in order to assess its reliability. The experimentally determined tension and distortion status at the acetabular bone-metal armour interface, lead to the wear and tear phenomenon, which can be explained by three mechanisms and theories incompletely reflecting the overall process. The histopathologic study of the acetabular bone tissue using FEM (finite elements method) on surgically removed specimens will probably lead to the identification of a series of factors that could reduce the rate of the wear and tear process.