Prognostic values of detecting MSI phenotypes in colorectal carcinoma by immunohistochemical method compared to molecular investigation.

BACKGROUND: The identification essentially of hMSH2 and/or hMLH1 alterations has clinical implications for recognition and prognosis of MSI phenotypes cases. In this study, we tried to identify instability by immunohistochemical expression pattern analysis, compared the results with molecular investigation and shown their usefulness as predictive factors for determination of Microsatellite Instability in patients with colorectal carcinomas in routinely. METHODS: Forty seven colorectal cancers and their adjacent colonic mucosa were selected retrospectively for this study. We first studied the potential value of molecular investigation to identify microsatellite instability in which a NCI panel (or Bethesda panel) of five microsatellite was analyzed (Bat-25, Bat-26, D2S123, D5S346 and D17S250). Secondary, we evaluated the immunohistochemical assessment of hMLH1, hMSH2, hMSH6 and PMS2 proteins in tumor and adjacent normal colorectal mucosa tissues. RESULTS: Fourteen cases were scored as MSI and the remaining MSS. Moreover, we found loss of expression for hMLH1, hMSH2, hMSH6 and PMS2 respectively in 9, 10, 6 and 9 of cases. The MSI patients were less than 45 years old, have right localization and mucinous histological type. We found an association between MSH2, age (P=0.03) and staging (P=0.02). MLH1 is associated only with age (P=0.02) while MSH6 with tumor grade (P=0.01). CONCLUSIONS: We found an association between MSI molecular investigation and MMR immunohistochemical expression which may allow one to specifically identify MSI phenotype of patients with colorectal carcinomas. Furthermore, immunohistochemical analysis of MMR protein can be used in routinely for detection of microsatellite instability without occurs to molecular investigation.

Uncommon etiology of knee pain: Lipoma arborescens.

 Lipoma arborescens is a rare intra-articular benign tumor. It affects mostly the knee and the diagnosis is usually difficult due to resembling symptomatology of osteoarthritis. CASES PRESENTATION: We report herein 3 new cases of lipoma arborescens of the knee in which a patient has synovitis associated. In 2 cases, the discovery was intraoperatively. We also report a tumor relapse 2 years after surgery.

Light-chain deposition disease of the kidney: a case report.

A 41-year-old man was admitted for evaluation of nephrotic syndrome associated with microhematuria, hypertension, and moderate renal failure. In serum and urine samples, monoclonal IgG-lambda was detected. Bone marrow examination showed normal representation of all cell lines with normal range of plasma cells. Renal biopsy demonstrated diabetes-like nodular glomerulosclerosis. Immunofluorescence failed to demonstrate the presence of kappa or lambda light chains in the kidney. Electron microcopy showed granular electron-dense deposits along the glomerular basement membranes and in the mesangial nodules. The patient was diagnosed as having light-chain deposition disease (LCDD) without evidence of plasma cell dyscrasia. This report was designed to stress the significant challenges that remain in the diagnosis of LCDD-related glomerulopathy. The salient morphological features that help in making an accurate diagnosis are discussed.

Microscopy and microanalysis study of the indium (In) behavior in the intestinal mucosa, the liver, the kidney and the testicle.

Several studies have demonstrated that In used in medicine has several impacts on organs like spleen and lungs after its systemic administration. In the present study, ultrastructural and microanalytical methods were used to investigate the impact of the presence of this element in the intestinal mucosa, the liver, the kidney and the testicle after its administration in two ways. After intraperitoneal administration, In was selectively concentrated in the lysosomes of hepatocytes, of tubular proximal convoluted cells and of Sertoli and Leydig cells. After intragastric administration, ultrastructural study showed that this element was concentrated in the lysosomes of duodenal enterocytes. Microanalytical methods showed that In was precipitated in those organelles in the form of insoluble phosphate salts. Similarly to other studies, it seemed that since In is a foreign element for the organism, it was precipitated in lysosomes, very probably due to the activity of an intralysosomal enzyme the acid phosphatase, to avoid its invasion to organism via the blood. This mechanism of precipitation of the mineral elements is of great interest in the process of defensive reaction of the organism against intoxication by foreign elements.

Clinicopathological characteristics of obesity-associated focal segmental glomerulosclerosis.

Obesity-related glomerulopathy (ORG) is a secondary form of focal segmental glomerulosclerosis (FSGS) occurring in obese patients with a body-mass index higher than 30 kg/m(2). It is typically manifested by nephrotic-range proteinuria without full nephrotic syndrome, and progressive renal insufficiency. Characteristic morphologic features include the consistent presence of glomerulomegaly, predominance of perihilar variant of FSGS, and the relatively mild fusion of visceral epithelial cell foot processes. The concept of podocyte depletion as a driver of the glomerular scarring in obesity-associated FSGS is well documented. The underlying mechanisms are likely to be related in part to the oxidative stress and the impairment of the integrity of the slit diaphragm and cell adhesion resulting mainly from angiotensin II and transforming growth factor-ß. These proapoptotic cytokines are upregulated in obesity in response to insulin resistance, compensatory hyperinsulinemia and glomerular hyperfiltration-hypertension mediated mechanical stress. This review is designed to discuss the clinicopathologic features of obesity-associated FSGS, with a focus on the podocyte injury, which is involved in the onset and progression of the glomerulosclerotic process. Ultrastructural glomerular lesions are documented.

Membranoproliferative glomerulonephritis with isolated C3 deposits: case report and literature review.

Membranoproliferative glomerulonephritis with isolated C3 deposits (MPGNC3) is an uncommon condition characterized by overt glomerular C3 deposits in the absence of immunoglobulins and intramembranous dense deposits. Here the authors describe the clinical and morphological features of primary MPGNC3 in a 13-year-old boy and critically review the previously published cases. The patient presented with nephrotic syndrome and microscopic hematuria. Blood tests revealed very low circulating C3 levels. The renal biopsy exhibited subendothelial, subepithelial, and mesangial deposits, with C3 but not immunoglobulins seen on immunofluorescence. This case and the review of the literature indicate that the serum complement profile with decreased levels of C3 and normal levels of classical pathway components together with glomerular deposits containing exclusively complement C3 is highly suggestive of alternative pathway activation. The diagnosis of acquired and/or genetic complement abnormalities in some cases supports that complement dysregulation is implicated in the pathogenesis of MPGNC3. Such data show great promise to provide new therapy strategies based on modulation of the complement system activity.

[Kindler syndrome: clinical and ultra-structural particularities, a propos of three cases]. / Le syndrome de Kindler, particularités cliniques et ultrastucturales à travers trois nouvelles observations familiales.

INTRODUCTION: Kindler's syndrome is a rare type of genetic skin condition belonging to the class of bullous poikilodermia. We report three new sibling cases of this rare syndrome. CASES REPORTS: The condition was seen in three sisters aged 12, 16 and 20 years, born of a first-degree consanguineous marriage with no family history of Kindler's syndrome. The three patients presented spontaneously regressive bullous eruptions, poikilodermia of gradual onset, major cutaneous atrophy on the back of the hands and the feet, photosensitivity and gingival hypertrophy. Electron microscopy examination of poikilodermic skin showed normal anchoring filaments and intraepidermal cleavage. DISCUSSION: Diagnosis of Kindler's syndrome is based upon clinical evidence. Kidler's syndrome is a well defined clinical entity. Ultra-structural studies show intraepidermal, junctional, and dermal cleavage. This syndrome must be differentiated from congenital epidermolysis bullosa, Weary's syndrome, and other bullous hereditary poikilodermas.

Value of electron microscopy in the diagnosis of glomerular diseases.

To evaluate the contribution of electron microscopy to the final diagnosis of glomerulopathies, the authors established a prospective study during the first semester of 2006. A total of 52 kidney biopsies were performed with 3 samples for light microscopy, immunofluorescence, and electron microscopy. Among these renal biopsies, only 20 were examined with electron microscopy because the diagnosis made on the basis of conventional methods had remained unclear or doubtful. In 18 cases, electron microscopy was undertaken for the investigation of primary kidney disease. The 2 remaining cases were transplant biopsies. In this series of 20 patients, there were 3 children with an average age of 9 years and 17 adults with an average age of 35.5 years. Fifteen patients (75%) were nephrotic. The study revealed that electron microscopy was essential for diagnosis in 8 cases (40%) and was helpful in 12 cases (60%). In conclusion, the results showed that the ultrastructural study provides essential or helpful information in many cases of glomerular diseases, and therefore electron microscopy should be considered an important tool of diagnostic renal pathology. As was recommended, it is important to reserve renal tissue for ultrastructural study unless electron microscopy can be routinely used in all biopsies. Thus, this technique could be performed wherever a renal biopsy has to be ultrastructurally evaluated.

Msx and dlx homeogene expression in epithelial odontogenic tumors.

Epithelial odontogenic tumors are rare jaw pathologies that raise clinical diagnosis and prognosis dilemmas notably between ameloblastomas and clear cell odontogenic carcinomas (CCOCs). In line with previous studies, the molecular determinants of tooth development-amelogenin, Msx1, Msx2, Dlx2, Dlx3, Bmp2, and Bmp4-were analyzed by RT-PCR, ISH, and immunolabeling in 12 recurrent ameloblastomas and in one case of CCOC. Although Msx1 expression imitates normal cell differentiation in these tumors, other genes showed a distinct pattern depending on the type of tumor and the tissue involved. In benign ameloblastomas, ISH localized Dlx3 transcripts and inconstantly detected Msx2 transcripts in epithelial cells. In the CCOC, ISH established a lack of both Dlx3 and Msx2 transcripts but allowed identification of the antisense transcript of Msx1, which imitates the same scheme of distribution between mesenchyme and epithelium as in the cup stage of tooth development. Furthermore, while exploring the expression pattern of signal molecules by RT-PCR, Bmp2 was shown to be completely inactivated in the CCOC and irregularly noticeable in ameloblastomas. Bmp4 was always expressed in all the tumors. Based on the established roles of Msx and Dlx transcription factors in dental cell fates, these data suggest that their altered expression is a proposed trail to explain the genesis and/or the progression of odontogenic tumors.

An Immunoscore System Based On CD3+ And CD8+ Infiltrating Lymphocytes Densities To Predict The Outcome Of Patients With Colorectal Adenocarcinoma.

PURPOSE: The aim of this study was to evaluate the Immunoscore (IS) methodology as a prognostic marker of colorectal adenocarcinoma in Tunisian population. Tumor blocks were retrospectively collected from 106 patients with sporadic colorectal cancer. METHODS: Immunohistochemical staining and images analysis software were used to quantify the density of CD3+ and CD8+ tumor-infiltrating lymphocytes in the center of the tumor and invasive margin. RESULTS: The density of CD3+ and CD8+ was significantly associated with 5-year overall survival (P=0.001 and P=0.00098, respectively) and 5-year disease-free survival (P=0.0006 and P=0.0056, respectively). The earlier stage and the absence of vascular emboli showed a significant association with IS analysis. Cox multivariate regression analysis revealed that Immunoscore (from I0 to I4) was more significantly correlated with overall survival (P=0.00011) and disease-free survival (P=0.0008) than Tumor-Node-Metastasis (TNM) staging (P=0.057 and P=0.039, respectively). Patients with low IS were associated with inferior disease-free survival and overall survival, contrary to patients with high IS. CONCLUSION: This is the first study which evaluated the prognostic value of IS methodology in colorectal cancer in African and Arabic population. The IS methodology carries out in this study allows to estimate the risk of relapse in patients with colorectal cancer. Overall, our results support the implementation of the consensus Immunoscore as a new component for the classification of cancer, designated TNM-Immune.

Effects of the hydro-ethanolic extract of Marrubium vulgare in female rats.

OBJECTIVE: To evaluate the effects of ethanol-water (80:20) extract of Marrubium vulgare (M. vulgare) on the hematological parameters, macroscopic and histological aspects of the uterus and fetus in non-pregnant and pregnant rats. METHODS: Female rats were divided into 4 equal groups (n = 9), group N (normal rats) and group G (pregnant rats) considered as control groups, group NE (normal rats treated with the ethanol-water (80:20, v/v) extract of M. vulgare) and group GE (pregnant rats treated with the extract). The ethanol-water (80:20) plant extract was administered in a single daily dose 1 g/kg at the morning, during 19 d. On the 19 day of the experiment, animals were sacrificed, the uterus and fetuses were removed for the morphological and histological studies and the blood was collected in EDTA tubes for the measurement of hematological parameters with the use of an automate 'HORIBA ABX Micros 60 Hematology Analyzer'. RESULTS: Our results showed, in group NE and GE, a significant decrease on hematological parameters: red blood cells (NE: 18.6%; GE: 38.4%), hematocrit (NE: 13.8%; GE: 20.4%), hemoglobin (NE: 12.1%; GE: 8.3%) and mean corpuscular volume (NE: 6.4%; GE: 2%) with P more less a 0.05. Indeed, the extract of M. vulgare caused a significant decrease on the mean implantations of fetuses (82.5%, P < 0.001) and their size (47.2%, P < 0.01). As for the macroscopic and histological appearance of uterus, our data showed no change in normal treated rats. In contrast, the treated pregnant rats showed a severe histological change characterized by the existence of location of stopped gestation. Furthermore, it was also found in the uterus of these rat lyses placental and embryo tissue. CONCLUSIONS: All these results support the hypothesis of an abortifacient effect of M. vulgare.

Chromosomal evaluation in a group of Tunisian patients with non-obstructive azoospermia and severe oligozoospermia attending a Tunisian cytogenetic department.

Male infertility is the cause in half of all childless partnerships. Numerous factors contribute to male infertility, including chromosomal aberrations and gene defects. Few data exist regarding the association of these chromosomal aberrations with male infertility in Arab and North African populations. We therefore aimed to evaluate the frequency of chromosomal aberrations in a sample of 476 infertile men with non-obstructive azoospermia (n=328) or severe oligozoospermia (n=148) referred for routine cytogenetic analysis to the department of cytogenetics of the Pasteur Institute of Tunis. The overall incidence of chromosomal abnormalities was about 10.9%. Out of the 52 patients with abnormal cytogenetic findings, sex chromosome abnormalities were observed in 42 (80.7%) including Klinefelter syndrome in 37 (71%). Structural chromosome abnormalities involving autosomes (19.2%) and sex chromosomes were detected in 11 infertile men. Abnormal findings were more prevalent in the azoospermia group (14.02%) than in the severe oligozoospermia group (4.05%). The high frequency of chromosomal alterations in our series highlights the need for efficient genetic testing in infertile men, as results may help to determine the prognosis, as well as the choice of an assisted reproduction technique. Moreover, a genetic investigation could minimize the risk of transmitting genetic abnormalities to future generations.

Comparison of the intracellular behavior of gold (Au) and indium (In) in testicle after their parenteral administration.

The subcellular behavior of several mineral elements was studied using modern techniques of observation like transmission electron microscopy and analysis like electron probe microanalysis and secondary ion mass spectrometry. In the present ultrastructural and analytical investigations, we undertake to compare the intracellular behavior of a heavy metal, gold, and a III-A group element, indium, on rat testicular tissues after their parenteral administrations. Our ultrastructural results showed that while gold was found only in the lysosomes of Leydig cells under electron dense needles, indium was observed as electron-dense deposits in the lysosomes of both Leydig and Sertoli cells. No ultrastructural modifications were observed in the testicular tissues of the control rats. The microanalytical study showed that gold was concentrated in lysosomes with sulfur as a sulfate crystalline structure whereas indium was concentrated in the same organelle as insoluble phosphate salt. These results demonstrated that testicular Leydig and Sertoli cells have the ability to selectively concentrate indium but gold was concentrated only in the first kind of cells. The mechanism implicated in this concentration phenomenon is a biochemical one involving intralysosomal hydrolytic enzymes, the acid phosphatase and the arylsulfatase. This mechanism occurs in order to protect the organism and to avoid the presence of toxic metals under soluble and free form.

Role of parietal and principal gastric mucosa cells in the phenomenon of concentration of aluminum and indium.

The subcellular behavior of aluminum and indium, used in medical and industrial fields, was studied in the gastric mucosa and the liver after their intragastric administration to rats, using, two of the most sensitive methods of observation and microanalysis, the transmission electron microscopy, and the secondary ion mass spectrometry. The ultrastructural study showed the presence of electron dense deposits, in the lysosomes of parietal and principal gastric mucosa cells but no loaded lysosomes were observed in the different studied hepatic territories. The microanalytical study allowed the identification of the chemical species present in those deposits as aluminum or indium isotopes and the cartography of their distribution. No modification was observed in control rats tissues. In comparison to previous studies describing the mechanism of aluminum concentration in the gastric mucosa and showing that this element was concentrated in the lysosomes of fundic and antral human gastric mucosa, our study provided additional informations about the types of cells involved in the phenomenon of concentration of aluminum and indium, which are the parietal and the principal cells of the gastric mucosa. Our study demonstrated that these cells have the ability to concentrate selectively aluminum and indium in their lysosomes, as a defensive reaction against intoxication by foreign elements.

[Primary small intestinal lymphoma: epidemiological, histological and therapeutic transition in Tunisia]. / Transition épidémiologique, histologique et thérapeutique des lymphomes primitifs du grêle (LPG) en Tunisie.

INTRODUCTION: Primary small intestinal lymphoma (PSIL) is the second Non-Hodgkin lymphoma (NHL) of the digestive tract (after gastric NHL). PURPOSE: To evaluate during the past 28 years the epidemiological, anatomoclinical and therapeutic changes of PSIL in Tunisia through an acquired experience of more than a quarter of a century. METHODS: Our retrospective study included patients with histologically confirmed small intestinal lymphoma from 1981 to 2008 in Tunisia at Salah Azaiz Institute. The cohort of 210 patients was divided into two groups: A group from 1981 to 1992 (152 patients) and B group from 1993 to 2008 (58 patients). We analysed the epidemiological, anatomoclinical, histological, and therapeutic characteristics. RESULTS: We observed a significant decrease in the annual incidence of PSIL but also a significant transition of diffuse immunoproliferative small intestinal disease (IPSID) also known as "Mediterranean" PSIL, which were progressively replaced by "Western" lymphomas. Laparotomy with or without a debulking surgery, largely performed in group A, has disappeared at the cost of a primary chemotherapy (p < 0.001). Five-year actuarial global and relapse free survivals were respectively 60.5 and 57.3%. CONCLUSION: PSIL in Tunisia were subjected to a triple transition: epidemiological, histological and therapeutic.

Biosorption characteristics of phosphates from aqueous solution onto Phoenix dactylifera L. date palm fibers.

Phosphates are very important basic materials in agricultural and other industrial applications. The removal of phosphates from surface waters is generally necessary to avoid problems, such as eutrophication, particularly near urban areas. This paper is focused on the sorption of PO4(3-) ions from aqueous solution onto date palm fibers as a raw, natural and abundantly materials. A series of batch tests were conducted and the influence of contact time, initial phosphate concentration, pH of the solution and adsorbent dosage on PO4(3-) specie removal was investigated. FT-IR spectroscopy, scanning electron microscopy (SEM)/energy dispersive spectroscopy (EDS), transmission electron microscopy (TEM) analysis of the date palm fibers before and after phosphates biosorption and desorption studies were investigated to confirm the mechanism of the retention of phosphates. Results indicate that PO4(3-) uptake increased with increased initial phosphate concentration and decreased with increased pH values. The results showed that the highest phosphates adsorption capacity (4.35 mg/g) was found at pH 6.8, for an adsorbent dosage of 6g/L, initial phosphate concentration of 50mg/L, under a constant temperature of 18 degrees C+/-02, and the equilibrium state was reached within 120 min of exposure time. The relatively low cost and high capabilities of date palm fibers make them potentially attractive adsorbents for the removal of phosphate from aqueous solution.