Antigen Labeling Research Articles

Apoptosis and cell proliferation correlated with tumour grade in peritoneal fluids of patients with serous ovarian cancer

Apoptosis and cell proliferation in peritoneal fluids of patients with ovarian serous adenocarcinoma have not been well described in cytology. To investigate the contribution of cell death to the growth of this tumour we analysed both apoptosis and cell proliferation in peritoneal fluids of patients with ovarian serous adenocarcinoma. We studied 40 tumours from 40 patients with ovarian serous adenocarcinoma. Twelve tumours were high grade, 13 were moderately differentiated and 15 were poorly differentiated. The detection of DNA fragments in situ using the terminal deoxyribonucleotidy transferase (TDT)-mediated dUTP-digoxigenin nick-end labelling (TUNEL) assay was applied to investigate active cell death (apoptosis), and the MIB-1 antigen was used to investigate cell proliferation. The TUNEL indices were 0.29 ± 0.05, 0.79 ± 0.10 and 2.1 ± 0.90 in Grade I, Grade II and Grade III ovary carcinomas, respectively. The MIB-1 antigen labelling indices were 6.5 ± 0.09, 12.9 ± 3 and 25.8 ± 6.2, respectively, in the same order of tumour differentiation. The differences in both TUNEL and MIB-1 labelling indices were statistically significant between Grade I, Grade II and Grade III carcinomas and there was a positive correlation between the two indices (P < 0.001). Apoptosis and cell proliferation increased as the grade of tumour increased in ovarian serous adenocarcinoma, suggesting a rapid turnover of the tumour cells in tumours of higher grade, and may play an important role in the growth and the extension of such cancer cells in the peritoneal cavity.

Arginase-1

The distinction of hepatocellular carcinoma (HCC) from metastatic tumor in the liver often presents a diagnostic challenge that carries significant impact on prognostication and therapy. The number of diagnostically useful immunohistochemical markers of hepatocytes is limited to hepatocyte paraffin antigen (HepPar-1), polyclonal carcinoembryonic antigen, and CD10, with alpha-fetoprotein and glypican-3 labeling HCCs. Arginase-1 (Arg-1) is a binuclear manganese metalloenzyme that catalyzes the hydrolysis of arginine to ornithine and urea. We used immunohistochemistry to compare the sensitivity of Arg-1 to that of HepPar-1 in 151 HCCs. We found that the overall sensitivities of Arg-1 and HepPar-1 are 96.0% and 84.1%, respectively. The sensitivities of Arg-1 in well, moderately, and poorly differentiated HCCs are 100%, 96.2%, and 85.7%, respectively, whereas, in comparison, HepPar-1 demonstrated sensitivities of 100%, 83.0%, and 46.4% for well, moderately, and poorly differentiated tumors, respectively. There were no HCCs in our study that were reactive for HepPar-1 but nonreactive for Arg-1. We also examined Arg-1 expression in nonhepatocellular tumors, including many that are potential mimics of HCC (renal cell carcinomas, neuroendocrine tumors, melanomas, gastric adenocarcinomas, and adrenocortical carcinomas) and found that only 2 non-HCC tumors were reactive for Arg-1. Arg-1 represents a sensitive and specific marker of benign and malignant hepatocytes that may ultimately prove to be a useful diagnostic tool in routine surgical pathology practice.

Prevention of Cutaneous Tissue Contracture During Removal of Craniofacial Implant Superstructures for CT and MRI Studies

ABSTRACTObjectivesThe purpose of this study is to analyze the morphological change of cell nuclei and the change of proliferating activity of oral malignancy and epithelial dysplasia between before and after photodynamic therapy in order to predict recurrence.Material and methodsWe experienced 14 cases of oral squamous cell carcinoma, one case of verrucous carcinoma and seven cases of epithelial dysplasia treated by photodynamic therapy (PDT). The mean nuclear area (NA) and coefficient of variation of the nuclear area (NACV) of 100 nuclei per slide were calculated using computer-assisted image analysis in hematoxylin and eosin stained biopsy specimens before and after PDT. Additionally, proliferating cell nuclear antigen (PCNA) immunohistochemistry was carried out in each specimen.ResultsThe mean NA after PDT was significantly lower than that before PDT in the nonrecurrent group. However, there was no significant difference in mean NA before and after PDT in the recurrent group. There were no significance differences in NACV before and after PDT in either the nonrecurrent or recurrent group. Furthermore, the PCNA labelling indices of the specimens after PDT was significantly lower than that before PDT in both the nonrecurrent and the recurrent group.ConclusionsMean nuclear area in the biopsy specimen after photodynamic therapy is likely to be a predictive marker for the recurrence of oral squamous cell carcinoma or epithelial dysplasia subjected to photodynamic therapy, while coefficient of variation of the nuclear area and proliferating cell nuclear antigen labelling indices are less helpful in predicting the recurrence of such lesions.

Ginsenosides promote proliferation of granulosa cells from chicken prehierarchical follicles through PKC activation and up‐regulated cyclin gene expression

The effect of GS (ginsenosides) on proliferation of chicken GCs (granulosa cells) from prehierarchical SYF (small yellow follicles) was evaluated, and involvement of the PKC (protein kinase C) signalling pathway as well as mRNA expression of cyclins and CDK (cyclin-dependent kinase) were investigated. Whole SYF or GCs isolated from SYF were cultured in Medium 199 supplemented with 0.5% FCS (fetal calf serum). After 16 h, the cells were challenged with GS alone or in combination with PKC inhibitor H7 or activator PMA (phorbol 12-myristate 13-acetate) for 24 h in serum-free medium. Results showed that in both whole follicles and pure GCs monolayer culture system, GS (0.1-10 microg/ml) significantly increased the number of GCs in SYF in a dose-dependent manner, and this stimulatory effect was inhibited by H7, but enhanced by PMA. Meanwhile, the PCNA-LI (proliferating cell nuclear antigen labelling index) of GCs displayed similar changes with the cell number. Mechanism of GS action was further evaluated in cultured GCs separated from SYF. Western blot analysis showed that 10 microg/ml GS increased PKC translocation from cytoplasm to the plasma membrane of the GCs to become the active state. This effect was blocked by H7. Furthermore, GS up-regulated the expression of cyclin D1/CDK6 and cyclin E/CDK2 mRNAs in GCs; however, inhibition of PKC with H7 attenuated this stimulatory effect. These results indicated that GS could stimulate proliferation of chicken GCs through activated PKC-involved up-regulation of cyclin D1/CDK6 and cyclin E/CDK2 genes, subsequently promoting development of the chicken prehierarchical follicles.

Metformin suppresses azoxymethane‐induced colorectal aberrant crypt foci by activating AMP‐activated protein kinase

Metformin is widely used for the treatment of diabetes mellitus. Adenosine monophosphate-activated protein kinase (AMPK) is known to be activated by metformin and to inhibit the mammalian target of rapamycin (mTOR) pathway. The mTOR pathway plays an important role in the protein translational machinery and cell proliferation. We examined the effect of metformin on the suppression of colorectal carcinogenesis in chemical carcinogen-induced models. Seven-wk-old BALB/c mice were intraperitoneally (i.p.) injected with azoxymethane (AOM, 10 mg/kg) and then treated with or without metformin (250 mg/kg/d) for 6 wk (for the investigation of aberrant crypt foci [ACF] formation) or 32 wk (for polyp formation). We next investigated colonic epithelial proliferation using bromodeoxyuridine (BrdU) and the proliferating cell nuclear antigen (PCNA) labeling indices. Furthermore, to examine the indirect effect of metformin, the insulin resistance status and the serum lipid levels were assessed. Treatment with metformin significantly reduced ACF formation. The effect of metformin on colon polyp inhibition was relatively modest. No significant difference in body weight or glucose concentration was observed. The BrdU and PCNA indices decreased in mice treated with metformin. A Western blot analysis revealed that the phosphorylated mTOR, S6 kinase, and S6 protein levels in the colonic mucosa decreased significantly in mice treated with metformin. In conclusion, metformin suppresses colonic epithelial proliferation via the inhibition of the mTOR pathway through the activation of AMPK. As metformin is already used daily as an antidiabetic drug, it might be a safe and promising candidate for the chemoprevention of colorectal cancer.

Evaluación de tres métodos para la inactivación de coliformes y Escherichia coli presentes en agua residual doméstica, empleada para riego

&lt;p&gt;&lt;strong&gt; &lt;/strong&gt;&lt;strong&gt;Objective.&lt;/strong&gt; To describe a standardized flow cytometry protocol for the relative and absolute quantification of hematopoietic cell subpopulations from normal bone marrow, and to evaluate the expression of different lineage-specific cell markers with a reactivity associated to cell differentiation to be used as part of the routine quality control in cytometry laboratories. &lt;strong&gt;Materials and methods. &lt;/strong&gt;The immunophenotypical analysis of different cell subpopulations was done with samples from normal bone marrow using a panel of monoclonal and polyclonal antibodies useful in the characterization of acute leukemias with four different fluorescences, by means of a protocol that combines cell labeling of membrane and cytoplasm antigens. Expression analysis was done in terms of mean fluorescence intensity (MFI). Fluorescent beads at a known concentration were added for calculating the absolute count of cells. &lt;strong&gt;Results.&lt;/strong&gt; The antibody panel used allowed the identification and quantification of different normal leukocyte subpopulations of lymphatic and myeloid origin, including CD34+ stem cells and more differentiated cell populations in the granulocytic, monocytic, and erythroid cell lines. We established reference values for cell populations and cell marker expression ranges as part of routine quality control of cytometry laboratories. &lt;strong&gt;Conclusion.&lt;/strong&gt; Immunophenotypic patterns identified as well as absolute and relative reference values for the different normal leukocyte populations from bone marrow can be used by cytometry laboratories as a basis for establishing reference parameters in phenotypic analyses of hematologic neoplasia.&lt;strong&gt; &lt;/strong&gt;&lt;/p&gt; &lt;p&gt;&lt;strong&gt;Key words: &lt;/strong&gt;multiparametric flow cytometry, immunophenotype, hematologic neoplasia, normal bone marrow, reference values, quality control.&lt;/p&gt;&lt;br /&gt;

Mesenchymal stem cells enhance growth and metastasis of colon cancer

Recently, mesenchymal stem cells (MSCs) were reported to migrate to tumor stroma as well as injured tissue. We examined the role of human MSCs in tumor stroma using an orthotopic nude mice model of KM12SM colon cancer. In in vivo experiments, systemically injected MSCs migrated to the stroma of orthotopic colon tumors and metastatic liver tumors. Orthotopic transplantation of KM12SM cells mixed with MSCs resulted in greater tumor weight than did transplantation of KM12SM cells alone. The survival rate was significantly lower in the mixed-cell group, and liver metastasis was seen only in this group. Moreover, tumors resulting from transplantation of mixed cells had a significantly higher proliferating cell nuclear antigen labeling index, significantly greater microvessel area and significantly lower apoptotic index. Splenic injection of KM12SM cells mixed with MSCs, in comparison to splenic injection of KM12SM cells alone, resulted in a significantly greater number of liver metastases. MSCs incorporated into the stroma of primary and metastatic tumors expressed α-smooth muscle actin and platelet-derived growth factor receptor-β as carcinoma-associated fibroblast (CAF) markers. In in vitro experiments, KM12SM cells recruited MSCs, and MSCs stimulated migration and invasion of tumor cells through the release of soluble factors. Collectively, MSCs migrate and differentiate into CAFs in tumor stroma, and they promote growth and metastasis of colon cancer by enhancing angiogenesis, migration and invasion and by inhibiting apoptosis of tumor cells.

Abstract 5111: Metformin suppresses azoxymethane-induced colorectal carcinogenesis via activating AMP-activated protein kinase

Abstract Metformin is widely used in the treatment of diabetes mellitus. Previous epidemiological reports suggest metformin reduce cancer incidence. Adenosine monophosphate-activated protein kinase (AMPK) is known to be activated by metformin, and inhibit the mammalian target of rapamycin (mTOR) pathway. The mTOR pathway plays an important role for protein translational machinery and cell proliferation. We examined the effect of metformin on the suppression of colorectal carcinogenesis in chemical carcinogen-induced models. Seven-week-old BALB/c mice i.p. injected with azoxymethane (AOM, 10 mg/kg) were treated with metformin (250 mg/kg/day) or without, for 6 weeks (for aberrant crypt foci (ACF) formation) or for 32 weeks (for polyp formation). We investigated colonic epithelial proliferation using the bromodeoxyuridine (BrdU) and proliferating cell nuclear antigen (PCNA) labeling indices. Western blot analysis was performed to elucidate the mechanism which affected colorectal carcinogenesis. Furthermore, to examine the indirect effect of metformin, the status of insulin resistance and serum lipid levels were assessed. Treatment with metformin significantly reduced the colon polyp and ACF formation, but there was no significant difference in body weight or glucose concentration. BrdU and PCNA indices decreased in mice treated with metformin. Western blot analysis revealed that phosphorylated mTOR, S6 kinase and S6 protein levels significantly decreased in mice treated with metformin. In conclusion, Metformin suppressed colonic epithelial proliferation via inhibition of the mTOR pathway by activation of AMPK. Metformin is already used daily as an antidiabetic drug, so it may be a safer and promising candidate for the chemoprevention of colorectal cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5111.

Abstract 4255: Significance of mesenchymal stem cells in growth and metastasis of colon cancer

Abstract Background: Recently, mesenchymal stem cells (MSCs) are reported to migrate to tumor sites as well as sites of injury and to incorporate into tumor stroma, but the effects of interactions between MSCs and tumor cells and the mechanisms underlying these effects remain unclear. We examined the role of MSCs in the tumor micro-environment using an orthotopic colon cancer model and a model of liver metastasis from colon cancer. Materials and Methods: Human MSCs labeled with fluorescent dye were injected into tail veins of nude mice bearing KM12SM human colon cancer cells. KM12SM cells alone or KM12SM cells mixed with MSCs were transplanted into the cecal wall or spleen of nude mice. Differentiation of MSCs in primary colon tumors and metastatic liver tumors were also analyzed. Migration assay and invasion assay were performed to examine attraction between KM12SM cells and MSCs in vitro. Results: In in vivo experiments, systemically injected MSCs migrated to the stroma of orthotopic colon tumors and metastatic liver tumors. Orthotopic transplantation of KM12SM cells mixed with MSCs resulted in significantly greater tumor weight than did transplantation of KM12SM cells alone (p &amp;lt; 0.05). The survival rate was significantly lower in the mixed-cell group, and liver metastasis was seen only in this group (p &amp;lt; 0.001). Moreover, tumors resulting from transplantation of mixed cells had a significantly higher proliferating cell nuclear antigen labeling index (p &amp;lt; 0.01), significantly greater microvessel area (p &amp;lt; 0.01), and significantly lower apoptotic index (p &amp;lt; 0.001). Splenic injection of KM12SM cells mixed with MSCs resulted in a significantly greater number of liver metastases than did splenic injection of KM12SM cells alone (p &amp;lt; 0.01). MSCs incorporated into the stroma of primary and metastatic tumors expressed α-smooth muscle actin and platelet-derived growth factor receptor-β as carcinoma-associated fibroblast (CAF) markers. In in vitro experiments, KM12SM cells recruited MSCs, and MSCs stimulated migration and invasion of tumor cells through the release of soluble factors (p &amp;lt; 0.05, respectively). Conclusion: MSCs migrate and differentiate into CAFs in tumor stroma, and they promote growth and metastasis of colon cancer by enhancing angiogenesis and by inhibiting apoptosis of tumor cells. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4255.

Intestinal absorption and histomorphometry of Syrian hamsters ( Mesocricetus auratus) experimentally infected with Lawsonia intracellularis

The objective of this study was to evaluate the intestinal absorption and histomorphometry of hamsters experimentally infected with Lawsonia intracellularis and correlate these parameters with severity of infection based on immunohistochemistry. Sixty hamsters were equally divided into control and inoculated groups which were orally infected with intestinal mucosa homogenate from pigs naturally infected with L. intracellularis. The intestinal absorption of glucose, sodium, potassium and chloride was evaluated in live animals (25 inoculated and 25 control) on day 26 after inoculation. In this procedure, a standard solution was infused into the cranial jejunum and collected at the terminal ileum. The experimental infection was confirmed by gross and histopathological examination and L. intracellularis antigen labeling by immunohistochemistry. Histomorphometry analysis demonstrated positive correlation between intestinal crypt depth and severity of infection based on immunohistochemistry. Infected animals had significantly lower intestinal absorption of glucose, potassium and chloride. These results indicate a lower intestinal absorption as an important mechanism of diarrhea in hamsters experimentally infected with L. intracellularis. Therefore, malabsorption should be considered as the main mechanism involved in the physiopathology of the diarrhea in L. intracellularis infected animals.

Chemoprevention of 1,2-dimethylhydrazine-induced colonic preneoplastic lesions in Fischer rats by 6-methylsulfinylhexyl isothiocyanate, a wasabi derivative

The preventive effects of dietary exposure to a wasabi derivative 6-methylsulfinylhexyl isothiocyanate (6-MSITC) during the initiation and post-initiation phases on the development of 1,2-dimethylhydrazine (DMH)-induced colonic aberrant crypt foci (ACF), and β-catenin-accumulated crypts (BCAC) were investigated in male F344 rats. To induce ACF and BCAC, rats were given four weekly subcutaneous injections of DMH (40 mg/kg body weight). The rats also received diets containing 200 or 400 ppm 6-MSITC during the initiation or post-initiation phases. The experiment was terminated 12 weeks after the start. DMH exposure produced a substantial number of ACF (323.8±69.7/colon) and BCAC (3.80±1.05/cm(2)) at the end of the study. Dietary administration of 6-MSITC at a dose of 400 ppm during the initiation phase caused a significant reduction in the total number of ACF (52% reduction, P<0.0001), larger ACF (4 or more crypt ACF) (58% reduction, P<0.001) and BCAC (76% reduction, P<0.00001). The dietary exposure to 6-MSITC significantly reduced the size (crypt multiplicity) of BCAC during both initiation and post-initiation treatment when compared to group 1 treated with DMH alone. Immunohistochemically, 6-MSITC administration lowered the proliferating cell nuclear antigen labeling index in ACF and BCAC. In addition, protein levels of hepatic cytochrome P-450 isozymes at 24 h after 6-MSITC exposure were significantly suppressed (P<0.01). The results indicated that 6-MSITC exerted chemopreventive effects in the present short-term colon carcinogenesis bioassay, through alterations in cell proliferation activity and drug metabolizing enzyme levels.

Increased expression of Akt2 and activity of PI3K and cell proliferation with the ascending of tumor grade of human gliomas

Objective To study the expression of Akt2 and activation of PI3K in different grades of human gliomas and correlate the Akt2 expression with the proliferation activity of gliomas. Methods Akt2 expression and PI3K activity were examined in 48 different grades of human glioma specimens and six normal brain tissue samples by immunohistochemistry and Western blot analysis. The proliferation activity of tumors was evaluated by Ki-67 nuclear antigen labeling index (Ki-67 LI) using immunostaining. Results In contrast to the normal brain tissues, Akt2 expression and PI3K activity were greatly increased with the ascending of tumor grade and correlated positively with the proliferation activity of gliomas. Conclusion Akt2 may play a critical role in the development of gliomas and present a potential therapeutic target for malignant gliomas.

The Impact of the Intra-Abdominal Space on Liver Regeneration After a Partial Hepatectomy in Rats

Little is known about the relationship between intra-abdominal space and liver regeneration. The present study was experimentally designed to investigate the influence of the "occupied space" or the "loss of occupied space" on a regenerating liver. Experiment 1: Rats were randomly assigned to two groups: SO (space occupied) rats (n = 40); occupancy of intra-abdominal space followed by a two-thirds partial hepatectomy (PH) and control rats (n = 40); A PH alone. The rats in both groups were euthanized at 24, 48, 96, and 168 h after the operation. Computed tomography (CT) images were analyzed to evaluate the regenerating-direction and the shape of the regenerated remnant liver. The liver to body weight ratio and the proliferating cell nuclear antigen (PCNA) labeling index were measured at each time point. Experiment 2: A second laparotomy was performed at 168 h after the PH in both groups; occupier-removal for the SO rats and a sham operation for the control rats. The rats in both groups were euthanized at 24 and 168 h after the second operation. The liver to body weight ratio and PCNA labeling index were measured at each time point. Experiment 1: The remnant liver of the SO rats enlarged toward the dorsal and caudal side because liver regeneration toward the ventral side in the SO rats was inhibited with the occupier in the abdominal space at 96 h, and later, after the PH. CT images showed a statistically significant difference in the shape of the regenerated remnant liver between the control group and the SO group. The liver/body weight ratio was significantly decreased in the SO rats at 96 and 168 h after PH (P < 0.05). There was no significant difference between the groups in the PCNA labeling index. The SO rats showed a significant increase of the PCNA labeling of the inferior right lobe (10.6%) in comparison with the index of the superior right lobe (7.8%), which came in direct contact with the occupier, at 96 h after the operation (P < 0.05). The cell density of superior right lobe of the SO rats group was significantly higher than that of the control group at 168 h after operation (P < 0.05). Experiment 2: There was no statistically significant difference in the liver/body weight ratio at 168 hrs after the second operation between the groups. However, there was a statistically significant increase of the PCNA labeling index 24 h after the second operation in the occupier-removal rats in comparison with the control rats (P < 0.05). The occupied intra-abdominal space was therefore found to suppress liver regeneration after a partial hepatectomy, while the removal of such an occupied space stimulated the regeneration of the liver.

Increased Neuronal Proliferation in the Dentate Gyrus of Aged Rats Following Neural Stem Cell Implantation

It is now well accepted that the brain is able to generate newborn neurons from a population of resident multipotential neural stem cells (NSCs) located in two discrete regions of the brain. The capacity for neurogenesis appears to diminish over the lifespan of an organism. Methods to potentiate the proliferation of new neuronal or glial cells within the central nervous system from resident NSCs could have therapeutic potential following an insult, such as stroke, or to replace lost cells as a result of a neurodegenerative disease. We implanted cells from a human NSC cell line, CTX0E03, originally derived from fetal cortical tissue directly into the ventricles of aged rats. CTX0E03 cells have angiogenic properties via secretion of growth factors, so we investigated if the implanted cells would stimulate proliferation of NSCs within the subgranular zone (SGZ) of the dentate gyrus. Bromodeoxyuridine staining demonstrated significantly increased proliferation in the SGZ. Absence of double labeling for human nuclear antigen suggested that the increased proliferation was from endogenous neural progenitor cells. The acute treatment also led to an increased number of immature neurons as demonstrated by immunohistochemical staining for the immature neuronal marker doublecortin. The data suggest that implants of exogenous NSCs may promote regeneration in aging organisms through stimulation of endogenous neurogenesis.

Correlation of microvascular density and clinicopathological factors in different subtypes of renal cell carcinoma

The aim of this study was to evaluate the expressions of CD34, CD31 and microvessel density (MVD) in different subtypes of renal cell carcinoma (RCC) as well as the relationship between MVD and clinicopathological factors. Expressions of CD31 and CD34 were detected in 149 patients with RCC using SP immunohistochemical staining. The MVD was studied by Weidner's method. The expressions of CD31 and CD34 in the clear cell renal cell carcinoma (CCRCC) (98.35 +/- 55.05, 128.04 +/- 46.44) were significantly higher than those in chromophobe renal cell carcinoma (ChRCC) (30.70 +/- 17.72, 48.55 +/- 14.09) and papillary renal cell carcinoma (PRCC) (21.60 +/- 9.38, 38.12 +/- 10.98) (P < 0.01). The MVD value marked by CD31 (30.70 +/- 17.72, 21.60 +/- 9.38) was much lower than that marked by CD34 (48.55 +/- 14.09, 38.12 +/- 10.98) between ChRCC and PRCC (P < 0.01). Smaller and immatured microvessels and even single endothelial cells could be clearly seen. The MVD values marked by CD31 and CD34 were negatively correlated with the pathological grades (r(CD34) = -0.618, P < 0.01; r(CD31) = -0.442, P < 0.01) and clinical stages (r(CD34) = -0.283, P < 0.05; r(CD31) = -0.256, P < 0.05) in CCRCC. But no association was found in non-CCRCC (P > 0.05). MVD is significantly correlated with different types of endothelial labeling. The microvascular endothelial cells could be shown clearly by its related antigen labeling such as CD34 and CD31. CD34 is more sensitive than CD31. The MVD of CCRCC is significantly higher than that in non-CCRCC. The expressions of CD31 and CD34 are not correlated with tumor grade and stage in ChRCC and PRCC, while there is a negative correlation in CCRCC.

Clinicopathologic and immunohistochemical study of choroid plexus tumors: single-institution experience in Mexican population

In recent years, few studies have specifically focused on only histological features in choroid plexus tumors. We retrospectively reviewed the clinicopathologic and histological features in 37 patients with choroid plexus tumors and correlated these with glial fibrillary acidic protein (GFAP) expression and proliferation cell nuclear antigen (PCNA), p53, p21, and Rb labeling indexes, with special attention to tumor recurrence/regrowth. The study included 24 choroid plexus papillomas (CPPs), 4 atypical choroid plexus papillomas (ACPPs), and 9 choroid plexus carcinomas (CPCs). Patient age ranged from 15 to 70 years (mean 44 years). Most of the choroid plexus tumors were located in the IV ventricle. Recurrence was observed in 21 (52%) cases, 14 of which were CPP and 7 of which were CPC (P = 0.032). Histologic findings included major necrosis, fibrosis and psammoma bodies, amyloid deposits, inflammation, and thick vessels in recurrent tumors. The PCNA labeling index was 52.04 + or - 13.92 in CPPs, 76.50 + or - 17 in ACPPs, and 95.22 + or - 21.34 in CPCs (P = 0.009), and 67.43 + or - 28 in recurrent tumors. Similar values were found for p53, p21, and Rb. Furthermore, we observed that these presented more histological changes, adding, than nonrecurrent tumors, as well as a higher proliferation index of cell-cycle markers, and these were dependent predictor factors of survival. Recurrent tumors showed a different biological behavior than nonrecurrent tumors, but histological observations showed no mitotic features in order to consider them as grade II.

Chemopreventive effects of mate against mouse mammary and colon carcinogenesis.

The present study evaluated the chemopreventive potential of mate tea-like intake on mammary and colon carcinogenesis initiated by 7,12-dimethylbenz(a)antracene (DMBA) and 1,2-dimethylhydrazine (DMH) in female Swiss mice. After the initiation period, the animals received basal diet and organic mate tea-like, conventional mate tea-like, or green tea (positive control) at 2.0% as the drinking fluid during 15 weeks. At week 20, colon and mammary gland were analyzed for preneoplastic and neoplastic lesions development. Colon and mammary gland complexes were processed for cell proliferation analysis, estimated by proliferating cell nuclear antigen labeling index (PCNA-LI%). Specially, organic mate tea-like reduced the values of PCNA-LI% in colonic crypts (p < .003) and in mammary glands (p < .05) in DMBA/DMH-initiated groups. A lower incidence of aberrant crypt foci (ACF) in colon (p = .03) and of hyperplastic and neoplastic lesions in mammary gland (p < .05 and p < .02, respectively) was observed in DMBA/DMH-initiated groups treated with organic mate tea-like. These results suggest that post-initiation treatment with organic mate tea-like inhibited the development of colon and mammary carcinogenesis in a two-step medium-term mouse carcinogenesis model.

Clear cell adenocarcinoma of the uterine cervix in a young pregnant woman: a case report with immunohistochemical study

The patient was a 30-year-old-pregnant woman (para 1, gradiva 1) without in utero exposure to diethylstilbestrol. A papillary and flat lesion was found by colposcopy in the uterine cervix in the routine examination at 10 pregnancy weeks. Biopsies showed papillary and tubular proliferation of malignant cells with clear glycogen-rich cytoplasm and malignant hobnail cells and were diagnosed as clear cell adenocarcinoma. The patient was treated by radical hysterectomy. The lesion of the cervix (10 × 15 × 5 mm) was clear cell adenocarcinoma with mild invasion into shallow myometrium (pTIb). An immunohistochemical study showed that the tumor cells were positive for pancytokeratins (AE1/3, CAM5.2, KL-1, and polyclonal wide), cytokeratin (CK) 7, CK8, CK18, CK19, epithelial membrane antigen, CA 125, p53 protein, and Ki-67 antigen (labeling = 95%). The tumor cells were negative for high-molecular weight CK (34βE12), CK5/6, CK14, CK20, vimentin, AFP, CEA, desmin, S100 protein, α-smooth muscle antigen, CD34, estrogen receptor, progesterone receptor, chromogranin, synaptophysin, neuron-specific enolase, and CD56. The patient is now of free of tumor. In the English literature, the present case is the second case of cervical clear cell adenocarcinoma during pregnancy, and the first case of cervical clear cell carcinoma investigating a wide range of immunoreactive antigens.

Expression of Nestin, Vimentin, and NCAM by Renal Interstitial Cells after Ischemic Tubular Injury

This work explores the distribution of various markers expressed by interstitial cells in rat kidneys after ischemic injury (35 minutes) during regeneration of S3 tubules of outer stripe of outer medulla (OSOM). Groups of experimental animals (n = 4) were sacrificed every two hours during the first 24 hours post-ischemia as well as 2, 3, 7, 14 days post-ischemia. The occurrence of lineage markers was analyzed on kidney sections by immunohistochemistry and morphometry during the process of tubular regeneration. In postischemic kidneys, interstitial cell proliferation, assessed by 5-bromo-2′-deoxyuridine (BrdU) and Proliferating Cell Nuclear Antigen (PCNA) labeling, was prominent in outer medulla and reach a maximum between 24 and 72 hours after reperfusion. This population was characterized by the coexpression of vimentin and nestin. The density of -Neural Cell Adhesion Molecule (NCAM) positive interstitial cells increased transiently (18–72 hours) in the vicinity of altered tubules. We have also localized a small population of α-Smooth Muscle Actin (SMA)-positive cells confined to chronically altered areas and characterized by a small proliferative index. In conclusion, we observed in the postischemic kidney a marked proliferation of interstitial cells that underwent transient phenotypical modifications. These interstitial cells could be implicated in processes leading to renal fibrosis.

A reproducible technique for specific labeling of antigens using preformed fluorescent molecular IgG‐F(ab′)2 complexes from primary antibodies of the same species

Immunolabeling two different antigens using the indirect approach with antibodies from the same species is not possible as secondary antibodies can bind to either primary target antibodies. In this study, we describe how preformed complexes of primary and secondary labeled antibodies can be used in such circumstances. In this situation, the first antigen is labeled using the conventional indirect method followed by incubation with the preformed primary-secondary antibody complex against the second antigen. To prevent unbound secondary antibody from binding the indirectly-labeled antibodies, resulting in a false positive, we quenched excess secondary antibody with nonimmune murine serum from the species of the primary antibody. Before the formation of the preformed complex, the optimum dilution of both primary and secondary antibodies was determined. Once these concentrations were established, the concentration of nonimmune murine serum required to quench excess unbound secondary was determined. This step was accomplished by first incubating the sample with an antibody against an antigen known to be localized away from the antigen of interest, followed by the preformed complex. If specific staining was seen, other than that expected from the preformed complex, then the concentration of the serum was deemed insufficient for quenching, and increased accordingly. We demonstrate that this approach is successful in determining the optimum conditions for the preformation of ascites and purified monoclonal primary IgG with fluorescently conjugated F(ab')(2). Double immunolabelling of two focal adhesion antigens and two cytoskeletal proteins, with two murine primary antibodies, are presented as examples of the methodology.