Detection of alternatively spliced EMR2 mRNAs in colorectal tumor cell lines but rare expression of the molecule in colorectal adenocarcinomas

EMR2 and CD97, members of the epidermal growth factor (EGF)-TM7 family, show a very high homology. CD97, whose expression is closely related to clinical tumor stage in colorectal carcinomas, potentially functions as an adhesion molecule. Nothing is known about the presence of EMR2 in these tumors. We systematically examined the expression of EMR2 in colorectal carcinoma cell lines and adenocarcinomas. Of 18 cell lines, 10 were only slightly positive for EMR2 according to flow cytometry. Various EMR2 splice variants, including a new isoform, have been detected at the mRNA level. EMR2 expression did not correlate with in vitro migration or invasion capacity of the cell lines. Normal colorectal epithelial cells were EMR2 negative. In contrast to CD97, which is found in most colorectal adenocarcinomas, only 8 of 81 of these tumors expressed EMR2. No correlation was found between EMR2 expression and clinicopathological parameters of the tumors. In summary, a significant but low number of colorectal carcinomas are positive for EMR2, indicating different roles for this molecule and CD97 in these tumors.     

Towards biomimetic scaffolds: anhydrous scaffold fabrication from biodegradable amine-reactive diblock copolymers

The development of biomimetic materials and their processing into three-dimensional cell carrying scaffolds is one promising tissue engineering strategy to improve cell adhesion, growth and differentiation on polymeric constructs developing mature and viable tissue. This study was concerned with the fabrication of scaffolds made from amine-reactive diblock copolymers, N-succinimidyl tartrate monoamine poly(ethylene glycol)-block-poly(D,L-lactic acid), which are able to suppress unspecific protein adsorption and to covalently bind proteins or peptides. An appropriate technique for their processing had to be both anhydrous, to avoid hydrolysis of the active ester, and suitable for the generation of interconnected porous structures. Attempts to fabricate scaffolds utilizing hard paraffin microparticles as hexane-extractable porogens failed. Consequently, a technique was developed involving lipid microparticles, which served as biocompatible porogens on which the scaffold forming polymer was precipitated in the porogen extraction media (n-hexane). Porogen melting during the extraction and polymer precipitation step led to an interconnected network of pores. Suitable lipid mixtures and their melting points, extraction conditions (temperature and time) and a low-toxic polymer solvent system were determined for their use in processing diblock copolymers of different molecular weights (22 and 42 kDa) into highly porous off-the-shelf cell carriers ready for easy surface modification towards biomimetic scaffolds. Insulin was employed to demonstrate the principal of instant protein coupling to a prefabricated scaffold.     

Plasmodium falciparum histidine-rich protein II binds to actin,phosphatidylinositol 4,5-bisphosphate and erythrocyte ghosts in a pH-dependent manner and undergoes coil-to-helix transitions in anionic micelles

The recombinant histidine-rich protein II (HRPII) from Plasmodium falciparum was shown to bind actin and phosphatidylinositol 4,5-bisphosphate (PIP(2)) in vitro in a pH-dependent manner, very similar to hisactophilin, an actin-binding protein from ameba. Binding of HRPII to actin and PIP(2) occurred at pH 6.0 and 6.5, but not above pH 7.0. Circular dichroism (CD) spectroscopy confirmed that HRPII interacts with actin at pH below 7.0, as judged by the changes induced in the secondary structure of the HRPII/actin mixture. Further CD analysis demonstrated that HRPII adopts a predominantly alpha-helical conformation with anionic micelles of PIP(2) and SDS, but not with neutral micelles of phosphatidylcholine (PC), a feature that is common to many actin-binding proteins involved in cytoskeleton remodeling. Similarly to hisactophilin, a GFP-HRPII fusion protein shuttled from the cytoplasm to the nucleus of HeLa cells as the cellular pH was lowered from 8.0 to 6.0. HeLa cells transfected with the HRPII gene showed increased levels of histidine-rich proteins (HRPs) in the soluble cell fraction at pH 8.0. At pH 6.0, however, HRPs were detected mainly in the insoluble cell fraction. Interestingly, we found that HRPII binds to human erythrocyte membranes at pH 6.0 and 6.5 but not at pH above 7.0. Our results point to remarkable similarities between HRPII, hisactophilin, and actin-binding proteins. Possible roles of the HRPII during Plasmodium infection are discussed in the light of these findings.     

Clinical and endocrine follow-up of patients after testicular sperm extraction

OBJECTIVE: To evaluate the risk of testicular damage from testicular biopsies that are carried out for testicular sperm extraction (TESE) in infertile men. DESIGN: Prospective controlled clinical study. SETTING: Academic hospital. PATIENT(S): Forty infertile males with azoospermia.Examination of the clinical, endocrine, biochemical, and sonographic data in average after 18 months after TESE was performed. MAIN OUTCOME MEASURE(S): Measurements before and after TESE: hormone values, testicular size, morphologic characteristics, and power Doppler after scrotal sonography. RESULT(S): Comparison of preoperative and postoperative values of basal testosterone, FSH, LH, and estradiol levels did not reveal any differences. Twelve of 26 patients had subnormal testosterone values before TESE; 14 of 39 patients had subnormal levels afterward. Postoperative sonographic measurements showed no significant difference of the testicular volume as compared with the preoperative values. Results of power Doppler sonography revealed pathological conditions (n = 5) in patients with former iliacal or testicular operations. CONCLUSION(S): Endocrine testicular function and testicular size were not impaired after testicular biopsy when compared with preoperative data. However, patients with nonobstructive azoospermia seem to be at risk for androgen deficiency due to primary testicular failure after repeated testicular biopsies.     

Treatment of a case of primary retroperitoneal mucinous cystadenocarcinoma: is adjuvant hysterectomy and bilateral salpingo-oophorectomy justified?

OBJECTIVE: We present a case of primary retroperitoneal mucinous cystadenocarcinoma in a 38-year-old woman. STUDY DESIGN: The tumor was resected with a segment of adjacent descending colon. Five years after the operation, the patient is well, without evidence of recurring disease, based on clinical investigation and modern imaging techniques. RESULTS: In the light of the literature, it appears most likely that this rare tumor is caused by coelomic metaplasia. On the basis of the histopathologic findings in our case and the reports from the literature, we recommend radical tumor excision en bloc with all infiltrated adjacent structures. CONCLUSION: Added removal of unaffected uterus and adnexes makes young women infertile and climacteric and is not yet validated by long-term results.