Successful Protein Extraction from Over-Fixed and Long-Term Stored Formalin-Fixed Tissues

One of the major breakthroughs in molecular pathology during the last decade was the successful extraction of full-length proteins from formalin-fixed and paraffin-embedded (FFPE) clinical tissues. However, only limited data are available for the protein extraction efficiency of over-fixed tissues and FFPE blocks that had been stored for more than 15 years in pathology archives. In this study we evaluated the protein extraction efficiency of FFPE tissues which had been formalin-fixed for up to 144 hours and tissue blocks that were stored for 20 years, comparing an established and a new commercial buffer system. Although there is a decrease in protein yield with increasing fixation time, the new buffer system allows a protein recovery of 66% from 144 hours fixed tissues compared to tissues that were fixed for 6 hours. Using the established extraction procedure, less than 50% protein recovery was seen. Similarly, the protein extraction efficiency decreases with longer storage times of the paraffin blocks. Comparing the two buffer systems, we found that 50% more proteins can be extracted from FFPE blocks that were stored for 20 years when the new buffer system is used. Taken together, our data show that the new buffer system is superior compared to the established one. Because tissue fixation times vary in the routine clinical setting and pathology archives contain billions of FFPE tissues blocks, our data are highly relevant for research, diagnosis, and treatment of disease

Slug enhances invasion ability of pancreatic cancer cells through upregulation of matrix metalloproteinase-9 and actin cytoskeleton remodeling

Slug, a member of the Snail family of transcription factors, has a crucial role in the regulation of epithelial-mesenchymal transition (EMT) by suppressing several epithelial markers and adhesion molecules, including E-cadherin. A recent study demonstrated that no relationship exists between Slug and E-cadherin in pancreatic cancer. Another study showed that in malignant mesothelioma effusions Slug was associated with matrix metalloproteinase (MMP) expression, but that there was no association with E-cadherin. F-ascin is an actin-bundling protein involved in filopodia assembly and cancer invasion and metastasis of multiple epithelial cancer types. In this study, we investigated Slug, E-cadherin, and MMP-9 expression using immunohistochemistry in 60 patients with pancreatic cancer and their correlation with carcinoma invasion and metastasis. Additionally, we observed the effects of Slug on invasion and metastasis in the pancreatic cancer cell line PANC-1. Alterations in Slug, MMP-9, and E-cadherin were determined by RT-PCR, western blot, and immunohistochemistry. Alterations in MMP-9 and F-actin cytoskeleton were determined by immunofluorescence staining, flow cytometry (FCM), or gelatin zymography. Slug, E-cadherin, and MMP-9 expression in pancreatic cancer was significantly associated with lymph node metastases and we found a significant correlation between Slug and MMP-9 expression; however, no significant correlation was observed between Slug and E-cadherin expression. Slug transfection significantly increased invasion and metastasis in PANC-1 cells and orthotopic tumor of mouse in vivo, and significantly upregulated and activated MMP-9; however, there was no effect on E-cadherin expression. Slug promoted the formation of lamelliopodia or filopodia in PANC-1 cells. The intracellular F-actin and MMP-9 was increased and relocated to the front of the extending pseudopodia from the perinuclear pool in Slug-transfected PANC-1 cells. These results suggest that Slug promotes migration and invasion of PANC-1 cells, which may correlate with the reorganization of MMP-9 and remodeling of the F-actin cytoskeleton, but not with E-cadherin expression

Glucagonoma as a rare case of neuroendocrine tumor of the pancreas: a case report

Introduction: Glucagonoma is a rare neuroendocrine tumor with characteristic features such as the presence of the glucagon-producing tumor, diabetes, and necrolytic migratory erythema. Case presentation: the 60-year-old patient was admitted to hospital with periodic pain in the right and middle upper abdominal appearing after eating. Laboratory tests presented a high glucose level and anemia. Tumor of the body and tail of the pancreas passing the organ pouch has been found during the surgery. Diagnosis of glucagonoma was confirmed in histopathological examination in immunohistochemical stainings: a positive reaction was observed with chromogranin, synaptophysin and CEA. The proliferative activity of Ki-67 was less than 1%. Staining for glucagon also was positive so confirmed the presence of neuroendocrine tumor - glucagonoma. Conclusions: Glucagonoma sometimes may occur without characteristic features which may cause delayed diagnosis. Early diagnosis of glucagonoma is important because it increases the chances of successful recovery

Physiotherapy in women after breast cancer treatment – review

Introduction: Breast cancer is the most common malignant tumor in women in the Polish region. Surgery is a basic method of breast cancer treatment. Surgery often carries a lot of unwanted changes as follows: limitation of mobility in the shoulder joint on the operated side, secondary lymphoedema, post mastectomy pain syndrome (PMPS), reduction of muscle strength or disorders in body posture. Therefore, the implementation of physiotherapeutic activities that are designed to prevent and eliminate postoperative complications seems very important. The main aim of this work was to present physiotherapeutic management in women after mastectomy based on the analysis of available literature. The physiotherapeutic process can be divided into three periods: early hospital, early ambulatory and the late ambulatory period. In the first period, active slow exercises, self-support of the upper limb on the operated side and breathing exercises on the thoracic track are used to prevent circulatory disorders, pulmonary complications, and edema. The early ambulatory period includes corrective exercises, general improvement exercises, stretching and learning of automatic massage of the upper limb of the operated side. The last period should be enriched by recreational methods of physical activity such as swimming, cycling or Nordic walking to maintain physical fitness, proper mobility of the shoulder girdle and improve the patient's psychophysical state. It is very important the patient regular continues the rehabilitation program after curing of breast cancer as well. In the case of secondary lymphoedema of the upper limb, comprehensive rehabilitation physiotherapy is used, consisting of manual lymphatic drainage, healing exercises, compression therapy, and skin care. Conclusions: Physiotherapy in women after breast cancer surgery is a complex and long-term process. Physiotherapeutic methods are effective in treating complications after surgery of breast cancer surgery. It is necessary to constantly update the physio-therapy knowledge in women after breast cancer surgery