Microsurgical Treatment for Intramedullary Spinal Cord Ependymomas: Radical Surgical Resection and Outcome.

May 2011 Azzazi, Alaa MD; Sakr, Sameh MD; Sedik, Mohamed MD Neurosurgery Quarterly Methods The study included 17 patients with spinal cord ependymomas; 7 men and 10 women with mean age of 37.211.7 years. Patients presented with pain, sensory disturbances, and motor dysfunction with a mean duration of symptoms of 12.48.4 months. McCormick clinical grading defined 3 grade I patients, 7 grade II patients, 5 grade III patients, and 2 grade IV patients. Preoperative magnetic resonance imagining showed well-demarcated intramedullary lesions with a median length of 2 vertebral bodies. Eight patients had cervical lesions, 2 patients had cervicothoracic and 2 patients had thoracic lesions, and 5 patients had conus medullaris lesions. All patients had surgical resection using the microsurgical technique through posterior approach and the excised specimens were sent for pathological examination. Postoperative magnetic resonance imagining evaluating residual tumors and clinical grading were performed. Gross total resection could be achieved in 10 patients (58.8%), subtotal in 5 patients (29.4%), and biopsy in 2 patients (11.8%). Clinical grading determined immediately and 3 months postoperative were significantly superior to preoperative grading. Histopathological examination of excised specimens showed low-grade ependymoma. Four patients underwent radiotherapy after surgery. No change of clinical grading was reported in 5 patients (3 were grade I and 2 were grade IV) and 7 patients, 6 grade II and 1 grade III showed improvement to grade I. However, 5 patients, 4 grade III and 1 grade II showed immediate postoperative deterioration to grades IV and III, respectively, but at 3 months after surgery, 4 patients were improved, 3 to grade III and 1 to grade II. At the end of the follow up, 10 patients (58.8%) had stabilized and 7 patients (41.2%) had improved neurological status with no permanent deterioration and no recurrence for totally resected lesions was detected and no disease or surgery-related mortality was reported. Total-subtotal resection of intramedullary ependymomas using the microsurgical technique is a feasible and safe management policy without causing permanent neurological deficits and transient immediate postoperative neurological deterioration showed improvement within 3 months. The site and the degree of infiltration of the lesion manifested as preoperative neurological status are important determinants for the outcome.

Suspension fluorescence in situ hybridization (S-FISH) combined with automatic detection and laser microdissection for STR profiling of male cells in male/female mixtures. 25 March 2009 Mado Vandewoestyne & David Van Hoofstat Int J Legal Med Methods Sample preparation, Fluorescence in situ hybridization, fluorescence scanning, segmentation and masking, laser pressure catapulting, DNA extraction, amplification and detection. Fresh saliva was collected from healthy male and female volunteers. Five hundred microliters of phosphate-buffered saline was added to 1 ml of each saliva sample. The cell suspension was then centrifuged at 800g for 7 min and the supernatant was discarded. The cell pellet was washed once with 1 ml of PBS. After cell counting, mixtures were prepared by combining male and female buccal cells in 1:10 and 1:50 ratios. A cell pellet from the saliva of a female volunteer aftercataglottis with a male volunteer was prepared in the same way. Additionally, the skin of a female volunteer was swabbed with wet sterile cotton swabs 10 min, 1 h, and 8 h, respectively, after it was licked by a male volunteer. The swabs taken after 10 min and 8 h were immediately agitated in 1 ml of PBS to release the cellular material. The swabs taken after 1 h were dried for 24, 48, and 72 h, respectively, before they were agitated in 1 ml of PBS. From all samples, a cell pellet was prepared as described above.

cDNA microarray analysis of bovine embryo gene expression profiles during the preimplantation period. 24 November 2005 Koichi Ushizawa et al. Reproductive Biology and Endocrinology Methods Animals and sample collection, Sample RNA preparation RNA extraction, T7-based linear amplification, cDNA microarray, cDNA microarray hybridization, Data normalization of the cDNA microarray, Cluster analysis of the cDNA microarray data, and Real-time RT-PCR analysis. Bovine embryos on days 7, 14 and 21, extra-embryonic membranes on day 28 and fetuses on days 28 were collected to represent early embryo, elongating embryo, pre-implantation embryo, post-implantation extra-embryonic membrane and fetus, respectively. Gene expression at these different time points was analyzed using our cDNA microarray. Two clustering algorithms such ask-means and hierarchical clustering methods identified the expression patterns of differentially expressed genes across pre-implantation period. Novel candidate genes were confirmed by real-time RT-PCR. In total, 1,773 individual genes were analyzed by complete k-means clustering. Comparison of day 7 and day 14 revealed most genes increased during this period, and a small number of genes exhibiting altered expression decreased as gestation progressed. Clustering analysis demonstrated that trophoblast-cell-specific molecules such as placental lactogens, prolactinrelated proteins (PRPs), interferon-tau, and adhesion molecules apparently all play pivotal roles in the preparation needed for implantation, since their expression was remarkably enhanced during the pre-implantation period. The hierarchical clustering analysis and RT-PCR data revealed new functional roles for certain known genes as well as novel candidate genes related to already established trophoblast-specific genes such as PLs and PRPs. A large number of genes in extra-embryonic membrane increased up to implantation and these profiles provide information fundamental to an understanding of extra-embryonic membrane differentiation and development. Genes in significant expression suggest novel molecules in trophoblast differentiation.

The implication of second-trimester amniotic fluid TNF-alpha, cytochrome C and cell death nucleosomes in the prediction of preterm labor and/or premature rupture of membranes. 28 April 2011 Puchner K., et al. PubMed This case-control study, comprising 360 women undergoing genetic amniocentesis and out of whom 38 delivered preterm and 18 out of the latter after PROM, the above apoptotic molecules were determined by ELISA. The 38 cases with PTL and 18 cases with PROM were matched for age with 38 and 18 respective controls delivering at term, and the levels of apoptotic molecules were compared. Cell death nucleosome levels were found to be significantly associated with preterm delivery. Specifically, for every unit increase in nucleosomes, women were on average 0.2% more likely to deliver preterm. In contrast, such an association was not found concerning the other two apoptotic molecules TNF-a and Cytochrome C. Second-trimester amniotic fluid cell death nucleosomes' levels are significantly associated with preterm delivery and could possibly serve as predicting markers.

Genetic Mapping and Physical Cloning of UVB Susceptibility Region in Mice. 11 May 2005 Mary E Handel-Fernandez, Iwao Kurimoto, J Wayne Streilein and Vladimir Vincek Journal of Investigative Dermatology Methods Induction and assay of contact hypersensitivity, UVB radiation, Isolation of P1 clones, Restriction fragment mapping, RNA isolation and reverse transcriptasePCR, Subcloning and sequencing. Utilizing the differential response to UVB-induced contact hypersensitivity in various mouse congenic strains we genetically and physically isolated the region containing the UVB susceptibility locus. The region is located on chromosome 17 within the class III region of the mouse H-2 complex. The region has its human homolog within the MHC complex on the human chromosome 6 and was extensively studied over the last several years as many diseases of immunologic and dermatologic relevance as well as of non-immunologic background have been associated with this region. A large part of the human region has been completely sequenced and searched for possible transcripts. The part proximal to the human Bat2 has not yet been sequenced but was extensively searched for genes by various investigators in human and mouse. Therefore, it is likely that most of the genes residing between the Bat5 and H-2D are identified and that very few, if any, are still hiding. The genetic data clearly locate the UVB susceptibility locus to the Bat5H-2Dregion. Of the 14 genes already mapped to the homologous human region, we found that all but two of them are present in the same location in the mouse. The genes that were not found in the mouse, IC7 and NB6, have not been well characterized in humans. Only a partial human sequence is available for the NB6gene and was shown to contain an Alu repeat. This type of repetitive DNA sequence represents 10% of human and primate genomic DNA but is not found in the murine genome. Alu repeats are rarely present in the coding regions of mRNA; therefore, there is reason to doubt whether the NB6gene codes for a functional protein. Of the IC7 gene, only a partial sequence has been reported, and this sequence has no homology to any known gene. The failure to find mouse counterparts of these genes by us and by the investigators that originally described them in humans suggests that the mouse sequences have significantly diverged from the human sequence. If so, this would make these genes unlikely candidates for the UVB susceptibility gene. Of the 12 genes that we found residing in the human Bat5H-2D interval, all of them are expressed in the mouse skin and a keratinocyte cell line. Supposedly that the UVB susceptibility gene will be somehow involved in immunologic processes in the skin, it is unlikely, although not impossible that Bat1,Bat2, Bat3, Bat4, and G1 are candidates for UVB-S gene. Bat1 gene encodes a ubiquitously expressed putative nuclear RNA helicase. The G1 gene is a calcium-binding adapter molecule with unknown function. Bat2 and Bat3 are proline-rich proteins with ubiquitin-like domains that are expressed in most of the tested organs and particularly high in testis. The function of Bat4 is unknown, but similar to Bat2 and Bat3, it is highly expressed in the testis. Others have stated that this expression pattern may suggest a role in cell proliferation or differentiation.