cDNA cloning and expression of vitamin-K Epoxide Reductase (VKOR) gene from orange spotted grouper Epinephelus coioides

The cDNA sequence encoding Vitamin-K Epoxide Reductase (VKOR) was constructed from hypothalamus and pituitary cDNA libraries of Orange spotted Grouper Epinephelus coioides. VKOR gene contains 610 nucleotides including the open reading frame (ORE) of length 399 bp. ORE was started with ATG codon at the position of 199 and terminated with a stop codon TAA of 585. ORE sequence had 128 amino acids and the predicted protein molecular weight is approximately 14.08 kDa (GenBank: FJ939281.1). Expression of VKOR gene in different tissues was further examined using one-step RT-PCR. RT PCR result revealed that, VKOR mRNA expression was detected only in liver, kidney, hypothalamus and pituitary. VKOR gene sequence of E. coioides aligned with related sequences by multiple sequence alignment by clustal X and phylogenetic tree was generated by Neighbor Joining method using Genious v 5.4. E. coioides VKOR had the highest identity and homology with the puffer fish, Tetraodon nigroviridis, Takifugu rubripes and Zebra fish Danio rerio. Based on the VKOR expression in the kidney, liver hypothalamus and pituitary of the seven year's old (sex reversal stage) E. coioides, it may be involved in sex reverse.The cDNA sequence encoding Vitamin-K Epoxide Reductase (VKOR) was constructed from hypothalamus and pituitary cDNA libraries of Orange spotted Grouper Epinephelus coioides. VKOR gene contains 610 nucleotides including the open reading frame (ORE) of length 399 bp. ORE was started with ATG codon at the position of 199 and terminated with a stop codon TAA of 585. ORE sequence had 128 amino acids and the predicted protein molecular weight is approximately 14.08 kDa (GenBank: FJ939281.1). Expression of VKOR gene in different tissues was further examined using one-step RT-PCR. RT PCR result revealed that, VKOR mRNA expression was detected only in liver, kidney, hypothalamus and pituitary. VKOR gene sequence of E. coioides aligned with related sequences by multiple sequence alignment by clustal X and phylogenetic tree was generated by Neighbor Joining method using Genious v 5.4. E. coioides VKOR had the highest identity and homology with the puffer fish, Tetraodon nigroviridis, Takifugu rubripes and Zebra fish Danio rerio. Based on the VKOR expression in the kidney, liver hypothalamus and pituitary of the seven year's old (sex reversal stage) E. coioides, it may be involved in sex reverse

Pertinent issues related to laparoscopic radical prostatectomy

PURPOSE: We describe the critical steps of the laparoscopic radical prostatectomy (LRP) technique and discuss how they impact upon the pertinent issues regarding prostate cancer surgery: blood loss, potency and continence. RESULTS: A major advantage of LRP is the reduced operative blood loss. The precise placement of the dorsal vein complex stitch associated with the tamponading effect of the CO2 pneumoperitoneum significantly decrease venous bleeding, which is the main source of blood loss during radical prostatectomy. At the Cleveland Clinic, the average blood loss of our first 100 patients was 322.5 ml, resulting in low transfusion rates. The continuous venous bleeding narrowed pelvic surgical field and poor visibility can adversely impact on nerve preservation during open radical prostatectomy. Laparoscopy, with its enhanced and magnified vision in a relatively bloodless field allows for excellent identification and handling of the neurovascular bundles. During open retropubic radical prostatectomy, the pubic bone may impair visibility and access to the urethral stump, and the surgeon must tie the knots relying on tactile sensation alone. Consequently, open prostatectomy is associated with a prolonged catheterization period of 2 - 3 weeks. Comparatively, during laparoscopic radical prostatectomy all sutures are meticulously placed and each is tied under complete visual control, resulting in a precise mucosa-to-mucosa approximation. CONCLUSION: The laparoscopic approach may represent a reliable less invasive alternative to the conventional open approach. Despite the encouraging preliminary anatomical and functional outcomes, prospective randomized comparative trials are required to critically evaluate the role of laparoscopy for this sophisticated and delicate operation

Laparoscopic radical prostatectomy: omitting a pelvic drain

PURPOSE: Our goal was to assess outcomes of a selective drain placement strategy during laparoscopic radical prostatectomy (LRP) with a running urethrovesical anastomosis (RUVA) using cystographic imaging in all patients. Materials and Methods: A retrospective chart review was performed for all patients undergoing LRP between January 2003 and December 2004. The anastomosis was performed using a modified van Velthoven technique. A drain was placed at the discretion of the senior surgeon when a urinary leak was demonstrated with bladder irrigation, clinical suspicion for a urinary leak was high, or a complex bladder neck reconstruction was performed. Routine postoperative cystograms were obtained. RESULTS: 208 patients underwent LRP with a RUVA. Data including cystogram was available for 206 patients. The overall rate of cystographic urine leak was 5.8%. A drain was placed in 51 patients. Of these, 8 (15.6%) had a postoperative leak on cystogram. Of the 157 undrained patients, urine leak was radiographically visible in 4 (2.5%). The higher leak rate in the drained vs. undrained cohort was statistically significant (p = 0.002). Twenty-four patients underwent pelvic lymph node dissection (8 drained, 16 undrained). Three undrained patients developed lymphoceles, which presented clinically on average 3 weeks postoperatively. There were no urinomas or hematomas in either group. CONCLUSIONS: Routine placement of a pelvic drain after LRP with a RUVA is not necessary, unless the anastomotic integrity is suboptimal intraoperatively. Experienced clinical judgment is essential and accurate in identifying patients at risk for postoperative leakage. When suspicion is low, omitting a drain does not increase morbidity