Simultaneous Interference with HER1/EGFR and RAC1 Signaling Drives Cytostasis and Suppression of Survivin in Human Glioma Cells in Vitro.

We have previously reported that combined inhibition of the epidermal growth factor receptor by erlotinib and of RAC1 by NSC23766 yielded a synergistic antiproliferative effect on established and primary cultured glioblastoma cells. The current study aimed at identifying the molecular mechanism. Staining for annexin V/PI or carboxyfluorescein succinimidyl ester was performed in order to determine the induction of apoptosis, necrosis or cytostasis in established and primary cultured glioblastoma cells. Moreover, expression of Ki-67 was determined by immunofluorescence, and the expression of cell cycle proteins was analysed by Western blot. Our data show that combined treatment with erlotinib and NSC23766 resulted in a reduced number of cell divisions, a significantly decreased Ki-67 expression, increased apoptosis and autophagy when compared to single agent treatments. On the molecular level, concomitant treatment with both agents resulted in a pronounced downregulation of cyclin D1, cyclin-dependent kinases 2, 4 and 6, as well as of survivin when compared to treatments with either agent alone. In conclusion, we demonstrate that combined treatment of human glioma cell lines in vitro with erlotinib and NSC23766 markedly inhibits cell division, induces apoptosis independent of caspase-3 activation and induces autophagy concomitant with suppression of survivin.

Hereditary hyperferritinemia cataract syndrome: clinical, genetic, and laboratory findings in 5 families.

BACKGROUND: The hereditary hyperferritinemia cataract syndrome (HHCS) is an autosomal dominant disorder characterized by high serum ferritin and early onset cataract. Mutations in the iron responsive element (IRE) within the 5' untranslated region of the L-ferritin (FTL) gene lead to constitutive L-ferritin synthesis resulting in hyperferritinemia. Bilateral cataract formation is caused by the intracellular accumulation of ferritin in the lens. PATIENTS: 4 children from unrelated families were referred for further exploration of hyperferritinemia which was detected during the diagnostic work-up of gastroenterological or hematological disorders. 1 patient was primarily referred for the investigation of bilateral cataract.Diagnostics included routine blood analysis, including complete blood count, iron status, liver and kidney parameters, a physical and an ophthalmological examination. Molecular genetic analysis of the FTL IRE was performed in 4 patients by PCR from genomic DNA and subsequent direct sequencing. RESULTS: All index patients presented with isolated hyperferritinemia without iron overload and had a positive family history for early onset cataract. Age at onset and disease severity varied between different families and among family members. Molecular genetic analysis revealed point mutations within the FTL IRE. CONCLUSION: In patients with hyperferritinemia but without any other sign of iron overload or inflammation HHCS should be considered to avoid complex and invasive procedures. Vice versa, in patients with familial inherited cataract the early serum ferritin measurement helps to avoid unnecessary diagnostics.

Parathyroid adenomectomy under local anesthesia with intra-operative monitoring of UcAMP and/or 1-84 PTH.

Because 80% of patients with primary hyperparathyroidism have a single adenoma and because most adenomas are now visualized by ultrasonography, we have attempted to remove these suspected single adenomas under local anesthesia with intra-operative monitoring of urinary cAMP (UcAMP) and 1-84 parathyroid hormone (PTH) serum levels. In the last 2 years, 45 patients (mean age 65 years) with primary hyperparathyroidism underwent surgery with local anesthesia when a single adenoma was strongly suspected by ultrasonography. Patients with equivocal or misleading ultrasonography, e.g., those with associated thyroid or multiglandular pathology and those who were non-cooperative, were excluded from this procedure. UcAMP and 1-84 PTH were determined prior to the incision, at the time of removal of the adenoma, and at regular intervals until 120 minutes after the operation. Results were available 45 min to 60 min after sampling for PTH and 60 min to 80 min for UcAMP. Forty-two adenomas were removed through a 2 cm to 3 cm skin incision in a mean time of 25 minutes, with no adverse effect, no morbidity, and minimal discomfort. The 42 patients were normocalcaemic on follow-up. The monitorings always predicted the success of the operation. In the 3 remaining patients, because the monitorings remained elevated at the end of the procedure, the patients underwent classical bilateral neck dissection under general anesthesia. This new approach can be safely accomplished with short operative time and hospital stay. The absence of general anesthesia is reassuring for the patients who are reluctant to undergo general anesthesia.(ABSTRACT TRUNCATED AT 250 WORDS)

[Excision under local anesthesia of parathyroid adenomas. Criteria of selection and control of effectiveness. 35 cases]. / Exérèse sous anesthésie locale des adénomes parathyroïdiens. Critères de sélection et contrôle d'efficacité. 35 observations.

Primary hyperparathyroidism caused by an adenoma that has been identified and localized by ultrasonography can be treated through a limited approach route, under local anaesthesia, provided the effect of excision is controlled by a perioperative assay of urinary cAMP or, preferably, of plasma parathormone level, and provided the contra-indications of this method are respected. Thirty-three out of 35 patients have been successfully operated upon by this method. In case of failure confirmed by laboratory tests, local anaesthesia was only a prelude to cervicotomy under general anaesthesia.

[Technique and results of lung transplantation in rats]. / Technique et résultats de la transplantation pulmonaire chez le rat.

Lung transplantation has recently emerged as an efficient therapy for a variety of endstage pulmonary diseases. Nevertheless, many problems remain unsolved. A simple and reproducible experimental model allows examination of some of the problems encountered. We report here our experience of lung transplantation in the rat. One hundred and thirty six lung transplantations have been performed. After the initial phase necessary to master the technique, an overall survival rate of 86% was achieved. Graft function, as assessed by chest X-rays and lung isotope scans, was satisfactory up to 4 months after transplantation. Lung transplantation in the rat is a reliable and reproductible experimental model allowing the development of research projects in this field.

[T lymphocyte populations of the intestinal mucosa in celiac disease in children. Immunohistochemical study]. / Populations lymphocytaires T de la muqueuse intestinale au cours de la maladie coeliaque chez l'enfant. Etude immunohistochimique.

In order to study the distribution of lymphocyte subpopulations in a pathologic intestinal mucosa, the authors, instead of using the classic method by counting the number of lymphocytes, present an original method permitting the exploitation of quantified data from labelled surface cells by texture analyser coupled with a computerized system. We investigated 25 children presenting with chronic diarrhea and villous atrophy and 5 control subjects. Fifteen of the 25 children had celiac disease (10 active with total villous atrophy and 5, celiac disease in remission with healing mucosa), 5 cow's milk protein intolerance with total or partial villous atrophy and 5, chronic diarrhea with partial villous atrophy. Immunohistochemical study with monoclonal antibodies was carried out on frozen sections using a three-step immunoperoxidase technique. Compared with the 5 controls, patients with food intolerance (celiac disease and cow's milk protein intolerance) showed a significant increase of T suppressor lymphocytes (p less than 0.01 and p less than 0.05) in the epithelium, whereas there were more T helper lymphocytes in the lamina propria (p less than 0.05 and p less than 0.01). Non-treated celiac disease was distinguished from treated celiac disease by a marked increase in intra-epithelial T cytotoxic-suppressors. These results suggest that T cytotoxic-suppressors may be the mediators of the lesions observed in celiac disease.

[Xenogeneic hyperacute rejection of the lung in rats]. / Rejet hyperaigu xénogénique du poumon chez le rat.

Xenotransplantation, with organs harvested from appropriate donor animals could be a potential solution to the shortage of donor organs in general and of lung grafts in particular. Unilateral orthotopic lung transplantation in the rat has been used to study the lung xenograft rejection in a discordant donor-recipient combination, i.e. guinea-pig to rat. With this microsurgical technique, a high post-operative survival rate has been reached. The good function of the grafts was assessed by serial chest x-ray, lung perfusion scintigraphy until several months after the transplantation. The survival of the xenografts has been then compared to that of allografts and isografts. Lung xenograft rejection in this combination has been very fast: 53 minutes. Macroscopic and histologic data displayed lung infarct with hemorrhagic edema.

Two statistical approaches to nuclei shape and size in a morphometric description of lymph node sections in non-Hodgkin's lymphoma.

An attempt is made to evaluate more clearly the potential contribution of quantitative nuclear profile shape and size measurements to lymph node section histologic description in 70 cases of non-Hodgkin's lymphoma (NHL). The area of nuclear profiles and five nonredundant and size-free shape indices were measured using the THECLA program on a Leitz Texture Analysis System (Leitz-TAS). Two statistical approaches were applied, known respectively as "parametric," (first statistical moments of variable distributions over samples of 200 nuclear profiles) and "nonparametric," which are percentages of nuclear profiles distributed into five "cytological" classes that are defined by shape: round (A), elongated (B), kidney shaped (C), irregular (D) and cleaved (E) nuclear profiles. Both statistical approaches provide proper overall discrimination of the eight histological categories identified with reasonable reliability by pathologists. Above all, the present report discusses the ability of a set of parametric and nonparametric variables to describe NHL cell populations, in an objective and meaningful way, according to nuclei shape. A method of synthesizing multidimensional correlations (CORICO program) is proposed in support of the discussion. Also, the specific descriptive power of each of the variables is described; in particular, it is concluded that there is a close link between the shape and size of the nuclear profiles of the cells.

Prognostic significance of a morphometric study of lymph node biopsies in chronic lymphocytic leukaemia.

In 18 chronic lymphocytic leukaemia (CLL) and 2 diffuse, well-differentiated lymphoma (DWDL) lymph node biopsies, morphometric results analysing nuclear area distribution curves by the skewness coefficient were compared to histological findings, clinical stage and survival. Lymphocytic proliferation was subclassified according to the number and the distribution of large lymphoid cells. All cases with a component of large lymphoid cells had a right deviation of the nuclear area distribution curve and a positive value of the skewness coefficient in contrast to those where lymphocytic proliferation was monomorphous. Good correlation was observed between histological findings, morphometric analysis and survival: most patients with a right deviation of nuclear area distribution curve and positive skewness coefficient are clinically stage C, or died within 2-44 months, in contrast to patients with a symmetric distribution of nuclear area curve who are clinically stage A or B and still alive. The results indicate that in CCL and DWDL the lymph node biopsy with quantitative analysis has prognostic significance.

An application of mathematical morphology to analysis of the size and shape of nuclei in tissue sections of non-Hodgkin's lymphoma.

Using principles from the theory of mathematical morphology, a semiautomatic analysis of the size and shape of cell nuclei on tissue sections was carried out on a Leitz Texture Analysis System (Leitz-TAS). The four parameters proposed here are more discriminatory than conventional shape evaluation by the form factor (FF), which is based on the ratio of perimeter squared to area. The parameters quantified, respectively, nuclear elongation (ND), narrow (R1) and wide (R2) irregularities, and the distribution of R1 and R2 along the nuclear contour (ID). The properties of these parameters were tested nucleus-by-nucleus on 24 nuclear models. The methodology was then illustrated by a study of lymph node nuclei in non-Hodgkin's lymphoma (NHL). Prior to analysis, 45 lymphomas were classified into five categories of nuclear size and shape according to the International Working Formulation (IWF). Two hundred nuclei were measured on each lymph node section. Statistical interpretation was based upon an analysis of the nuclear surface area on sections and upon the mean values of R1, R2, and ND, the standard deviations of R1 and R2, and the percentage of cleaved nuclei detected by ID. The mean value of R2 discriminated best between the two sets of populations with regular and irregular nuclear contours, respectively. Parameters R1, ND, and ID permitted the distinction of certain NHL cases among populations with irregular nuclei. Nuclear invaginations decreased in depth as the nuclear area became greater. The median surface area was well correlated to the IWF, and the skewness coefficient (third statistical moment of the nuclear surface area distribution) was related to the number of nuclear size or shape subpopulations.

Morphometric characterization of nuclei in non-Hodgkin's malignant lymphoma.

In addition to the nuclear area and a form factor, four morphometric parameters of nuclear shape (ID, R1, R2 and ND), obtained by the application of the principles of mathematical morphology, were used to characterize the nuclear contours in non-Hodgkin's malignant lymphomas. The values for each parameter were determined in 58 cases of non-Hodgkin's lymphoma categorized according to the Kiel and National Cancer Institute classifications. Small-cell, mixed and large-cell lymphomas could be distinguished on the basis of the mean nuclear area. The shape parameters R1, R2 and ID were efficient discriminators of the large centrocytic (cleaved-cell) lymphomas. Neither size nor shape factors could distinguish between centroblastic and immunoblastic tumors. The good correlation between the morphometric findings and the histopathologic categories suggest that morphometry may provide a quantitative and objective method for grading lymphomas.

Lymphadenopathy in patients at risk for acquired immunodeficiency syndrome. Histopathology and histochemistry.

Lymph node biopsies of 12 patients at high risk for acquired immunodeficiency syndrome (AIDS) with generalized lymphadenopathy (AIDS-related complex [ARC]) and seven controls with conventional lymph node hyperplasia were examined by light microscopy and immunohistochemical staining of frozen tissue. The immunohistochemical results were quantified by planimetric means. Our findings show that the T helper/T suppressor-cytotoxic (Th/Tsc) ratio in lymph nodes from ARC patients is significantly decreased with respect to controls and that this decrease precedes the change in the Th/Tsc ratio in peripheral blood. Our findings distinguish between lymphadenopathy from patients with ARC and other forms of hyperplasia; the relation to AIDS is discussed.