Indoxyl Sulfate Promotes Arterial Thrombosis in Rat Model via Increased Levels of Complex TF/VII, PAI-1, Platelet Activation as Well as Decreased Contents of SIRT1 and SIRT3

Patients suffering from chronic kidney disease (CKD) are at a 20-fold higher risk of dying due to cardiovascular diseases (CVDs), primarily thrombosis following vascular injury. CKD is connected with retention of uremic toxins, especially indoxyl sulfate (IS), which are currently considered as a non-classical CKD-specific risk factor for CVDs. The present study aimed to examine the effect of chronic exposure to IS on the hemostatic system and arterial thrombosis in a model without greater interferences from the uremic milieu consisting of additional uremic toxins. Forty-eight male Wistar Crl:WI (cmdb) rats were divided into three groups: one control group and two experimental groups, which were exposed to 100 or 200 mg/kg of b.w./day of IS in drinking water for a period of 28 days. The control group received water without IS. At the end of the experiment, the induction of arterial thrombosis was performed. We investigated the impact of IS on thrombosis incidence, kinetics and strength of clot formation, platelet activity, aortic contents of sirtuin (SIRT) 1 and sirtuin 3 (SIRT3), hemostatic system, cardiorespiratory parameters, biochemistry of plasma and urine as well as histology of the thrombus, kidney, and liver. Obtained data revealed that chronic exposure to IS promotes arterial thrombosis via increased levels of complex tissue factor/factor VII, plasminogen activator inhibitor-1 (PAI-1), platelet activation, as well as decreased aortic levels of SIRT1 and SIRT3. Therefore, we hypothesize that IS enhances primary hemostasis leading to augmented formation of platelet plug with increased amounts of fibrin and affects secondary hemostasis through the influence on plasma coagulation and fibrinolysis factors, which results in the increased kinetics and strength of clot formation. The findings described may contribute to a better understanding of the mechanisms leading to increased thrombotic events in patients with CKD with elevated levels of IS

Targeted Deletion of the Metastasis-Associated Phosphatase Ptp4a3 (PRL-3) Suppresses Murine Colon Cancer

Ptp4a3 (commonly known as PRL-3) is an enigmatic member of the Ptp4a family of prenylated protein tyrosine phosphatases that are highly expressed in many human cancers. Despite strong correlations with tumor metastasis and poor patient prognosis, there is very limited understanding of this gene family's role in malignancy. Therefore, we created a gene-targeted murine knockout model for Ptp4a3, the most widely studied Ptp4a family member. Mice deficient for Ptp4a3 were grossly normal. Fewer homozygous-null males were observed at weaning, however, and they maintained a decreased body mass. Although Ptp4a3 is normally associated with late-stage cancer and metastasis, we observed increased Ptp4a3 expression in the colon of wildtype mice immediately following treatment with the carcinogen azoxymethane. To investigate the role of Ptp4a3 in malignancy, we used the most commonly studied murine colitis-associated colon cancer model. Wildtype mice treated with azoxymethane and dextran sodium sulfate developed approximately 7-10 tumors per mouse in the distal colon. The resulting tumor tissue had 4-fold more Ptp4a3 mRNA relative to normal colon epithelium and increased PTP4A3 protein. Ptp4a3-null mice developed 50% fewer colon tumors than wildtype mice after exposure to azoxymethane and dextran sodium sulfate. Tumors from the Ptp4a3-null mice had elevated levels of both IGF1Rβ and c-MYC compared to tumors replete with Ptp4a3, suggesting an enhanced cell signaling pathway engagement in the absence of the phosphatase. These results provide the first definitive evidence implicating Ptp4a3 in colon tumorigenesis and highlight the potential value of the phosphatase as a therapeutic target for early stage malignant disease. © 2013 Zimmerman et al

TGF-beta receptor 2 downregulation in tumour-associated stroma worsens prognosis and high-grade tumours show more tumour-associated macrophages and lower TGF-beta1 expression in colon carcinoma: a retrospective study

<p>Abstract</p> <p>Background</p> <p>Histological phenotype and clinical behaviour of malignant tumours are not only dependent on alterations in the epithelial cell compartment, but are affected by their interaction with inflammatory cells and tumour-associated stroma. Studies in animal models have shown influence of tumour-associated macrophages (TAM) on histological grade of differentiation in colon carcinoma. Disruption of transforming growth factor beta (TGF-beta) signalling in tumour cells is related to more aggressive clinical behaviour. Expression data of components of this pathway in tumour-associated stroma is limited.</p> <p>Methods</p> <p>Tissue micro arrays of 310 colon carcinomas from curatively resected patients in UICC stage II and III were established. In a first step we quantified amount of CD68 positive TAMs and expression of components of TGF-beta signalling (TGF-beta1, TGF-beta receptors type 1 and 2, Smad 3 and 4) in tumour and associated stroma. Further we analyzed correlation to histological and clinical parameters (histological grade of differentiation (low-grade (i.e. grade 1 and 2) vs. high-grade (i.e. grade 3 and 4)), lymph node metastasis, distant metastasis, 5 year cancer related survival) using Chi-square or Fisher's exact test, when appropriate, to compare frequencies, Kaplan-Meier method to calculate 5-year rates of distant metastases and cancer-related survival and log rank test to compare the rates of distant metastases and survival. To identify independent prognostic factors Cox regression analysis including lymph node status and grading was performed.</p> <p>Results</p> <p>High-grade tumours and those with lymph node metastases showed higher rates of TAMs and lower expression of TGF-beta1. Loss of nuclear Smad4 expression in tumor was associated with presence of lymph node metastasis, but no influence on prognosis could be demonstrated. Decrease of both TGF-beta receptors in tumour-associated stroma was associated with increased lymph node metastasis and shorter survival. Stromal TGF-beta receptor 2 expression was an independent prognostic factor for cancer related survival.</p> <p>Conclusion</p> <p>Histological phenotype and clinical behaviour of colon cancer is not only influenced by mutational incidents in tumour cells but also affected by interaction of tumour tissue with inflammatory cells like macrophages and associated stroma and TGF-beta signalling is one important part of this crosstalk. Further studies are needed to elucidate the exact mechanisms.</p

Tumor budding as a new histological parameter in the metastasis of colorectal cancer

Introduction:The presence of tumor budding, i.e.,single cancer cells or a nest of poorly differentiated cells at the front of tumor invasion appears to be a new histopathological indicator of increased aggressiveness of colorectal carcinoma. Purpose: The aim of this work was a retrospective evaluation of the invasion front (tumorbudding, vascular invasion,and lymphocytic infiltration) in postoperative biopsies of patients with colorectal carcinoma and analysis of the 5-year survival. Materialsand methods:The study was based on the material received after surgical treatment of 164 patients with colon cancer. Tissue was obtained directly following tumor resection, fixed in 10% formaldehyde and embedded in paraffin blocks using a routine method by melting with paraffin at a temperature of 56º C. These samples were then routinely stained with haematoxylin and eosin and underwent a histopathological evaluation, with particular attention being paid to the invasion front of the tumor. The immunohistochemical expression of cytokeratin 20 was also evaluated using anti-human CK20 monoclonal antibody (clone Ks.20.8, Dako, Poland). Results: Tumor budding was found in 124 out of 164 patients. Statistical analysis showed a correlation between the presence of tumor budding TB and depth of invasion (pT), lymph node metastasis, distant metastasis, lymphocytic infiltration,and vascular invasion. The cumulative five-year survival correlated with the lack of tumor budding and vascular invasion, as well as a decrease in lymphocyticinfiltration

Spigelian hernia: a case report

Spigelian hernias constitute a minute fraction of all abdominal hernias. In this monography, we present a case report of this relatively seldom seen phenomenon which some general surgeons never get to see during their medical career

Whipple’s disease as a systemic infectious disease – a case presentation

Introduction: Whipple’s disease is a chronic systemic infectious disorder with Tropheryma whipplei as an etiologic agent, occurring rarely and affecting numerous organs and systems. The variety of symptoms and a non-typical course make it difficult to establish a proper diagnosis. Purpose: In this study, etiopathogenesis, diagnostics and treatment of Whipple’s disease were presented based on the case report of 60-year-old man diagnosed with Whipple’s disease. Case presentation: Persistent diarrhoea with weight loss, lymphadenopathy in the abdominal cavity and moderate microcytic anemia predominated in the clinical picture. Diagnosis was put based on the clinical picture and macroscopic assessment of the small intestine and the presence of macrophages filled with a PAS-positive substance in the lamina propria. To deepen diagnostics, samples collected were assessed showing macrophages with the damaged mucosa, containing numerous elongated micro-organisms whose ultrastructure corresponded to Tropheryma whipplei. The patient’s clinical conditions improved after antibiotic therapy. Conclusions: It is vital to remember about Whipple’s disease in patients with chronic diseases due to a non-specific clinical picture and difficulties in establishing a proper diagnosis. When the disease is diagnosed unequivocally, proper and effective antibiotic therapy should be instituted immediately

Vaginal biocoenosis examining comparing to exfoliative cervical cytology

Introduction: At present, the gynaecologists have been increasingly frequently switching from vaginal biocoenosis assessment towards cervical cytology results to obtain information on the type of infection. Exfoliative cervical cytology is a screening test for dysplastic intraepithelial lesions and ectocervical cancers. One should emphasize however that one of the four parts of the new Bethesda classification specifies such inflammatory lesions as: Trichomonas vaginalis, Candida, Actinomyces, Chlamydia, cellular changes consistent with HSV infection and changes of bacterial flora. The gynaecologists however may perform vaginal biocoenosis assessment individually and diagnose its abnormalities in a relatively short timeframe.Conclusions: Lack of 100% correlation between the vaginal biocoenosis test and cytological result according to the Bethesda system means that assessment of vaginal microflora in phase-contrast microscopy should not be abandoned. Purpose: To analyse the association between lesions revealed during vaginal biocoenosis assessment in correlation to lesions described in the studies dedicated to cytological assessment of ectocervical smear. Material and methods: The study group included 1991 female patients scheduled for the follow-up cytological screening in a gynaecological office. Patients underwent gynaecological examination covering external areas, colposcopy, vaginal pH measurement, sampling for vaginal biocoenosis assessment purposes and cytological sampling. Results: It was demonstrated that diagnostic conformity for Candida sp accounted for only 17.2%, changes of bacterial flora for only 4% and – in the case of Trichomonas vaginalis - for only 3.9%. According to observations, bacterial infections and candidiases have been more frequently diagnosed during vaginal biocoenosis examining comparing to cytological screening, whereas infections with Trichomonas vaginalis have been more frequently diagnosed in cytological screening. Conclusions: Lack of 100% correlation between the vaginal biocoenosis test and cytological result according to the Bethesda system means that assessment of vaginal microflora in phase-contrast microscopy should not be abandoned