Involvement of RARRES3 in the regulation of Wnt proteins acylation and signaling activities in human breast cancer cells.

The Wnt/ß-catenin signaling pathway has emerged as a key regulator of complex biological processes, such as embryonic development, cell proliferation, cell fate decision and tumorigenesis. Recent studies have shown that the deregulation of Wnt/ß-catenin signaling is frequently observed and leads to abnormal cell growth in human breast cancer cells. In this study, we identified a novel regulatory mechanism of Wnt/ß-catenin signaling through RARRES3 that targets and modulates the acylation status of Wnt proteins and co-receptor low-density lipoprotein receptor-related protein 6, resulting in the suppression of epithelial-mesenchymal transition and cancer stem cell properties. Mutation of the conserved active site residues of RARRES3 indicates that RARRES3 serves as an acyl protein thioesterase that tethers its target proteins and modulates their acylation status. Furthermore, the functions of p53 in cell proliferation and Wnt/ß-catenin signaling are significantly associated with the induction of RARRES3. Thus our findings provide a new insight into the molecular link between p53, protein acylation and Wnt/ß-catenin signaling whereby RARRES3 plays a pivotal role in modulating the acylation status of signaling proteins.

Resolution of the Acanthopagrus black seabream complex based on mitochondrial and amplified fragment-length polymorphism analyses.

In this study, DNA analyses were employed to verify the identity of six morphologically similar species that occur in the coastal waters of Taiwan: the black seabream complex (Acanthopagrus latus, Acanthopagrus schlegelii, Acanthopagrus sivicolus, Acanthopagrus taiwanensis, Acanthopagrus chinshira and Acanthopagrus pacificus). Amplified fragment-length polymorphism (AFLP) analyses clearly distinguished the same six species that are morphologically diagnosable based on subtle differences in scale counts and anal-fin colouration. In contrast, mitochondrial DNA analyses based on cytochrome b gene sequences did not distinguish individuals of A. schlegelii and A. sivicolus, reflecting either historical introgression or recent speciation and incomplete sorting of their mitochondrial lineages. Phylogenetic relationships among these six north-west Pacific Ocean species of Acanthopagrus analysed using AFLP data were consistent with scale rows above the lateral line (TRac), sperm ultrastructure and geographical distribution. The study provides molecular tools for future research relevant to improved management of these resources, and an increased understanding of the evolutionary history of this radiation.

Two spinefoot colour morphs: mottled spinefoot Siganus fuscescens and white-spotted spinefoot Siganus canaliculatus are synonyms.

Mottled spinefoot Siganus fuscescens and white-spotted spinefoot Siganus canaliculatus are two similar species that differ subtly in colouration and morphology. Three major mtDNA clades were identified for these species, but individuals were clustered by amplified fragment length polymorphism (AFLP) according to geography rather than morphology, suggesting that the colour morphs are interbreeding.

Localization of ubiquitin specific protease 26 at blood-testis barrier and near Sertoli cell-germ cell interface in mouse testes.

Ubiquitin Specific Protease 26 (USP26) is a little studied ubiquitin-specific protease that is expressed specifically in the testis. In humans, some USP26 polymorphisms have been reported to be associated with impaired male fertility. However, how USP26 affects male reproduction remains unclear. We generated an antibody that stained specifically cultured cells expressing an epitope-tagged USP26 and used it to elucidate the biological function of USP26. Immunostaining of mouse testis sections as well as dispersed germ cells showed the presence of USP26 at the blood-testis barrier, near the Sertoli cell-germ cell interface of post-step 7 spermatids, and coating the dorsal surface of sperm head. Further RT-PCR assays detected the expression of Usp26 in germ cells, but not in primary Sertoli cell lines. In addition, USP26 immunoprecipitated from testis lysates exhibited deubiquitinating activities. The localization of USP26 in the testis suggests a possible role in the movement of germ cells along the seminiferous epithelium.

Dystocia in Formosan sambar deer (Cervus unicolor swinhoei) in semi-domesticated herds.

Over a nine-year period (2000 to 2008), 22 instances of dystocia and 50 normal fawnings in Formosan sambar deer (Cervus unicolor swinhoei) hinds from semi-domesticated herds were investigated. The majority of the hinds (17, 77.3 per cent) that suffered dystocia were categorised as having a body condition score (BCS) in the medium range (between 2.5 and 3.5 inclusive), indicating that there was no significant relationship between obesity (BCS >3.5) in hinds and the occurrence of dystocia. In addition, maternal body condition had no effect on the birthweight of the offspring. Among the 22 instances of dystocia, 16 of the fawns (72.7 per cent) did not survive; one of these deaths was anteparturient and the other 15 were parturient. Eighteen (81.8 per cent) of the cases of dystocia involved male fawns, and 15 (93.8 per cent) of the deaths were of male fawns (P=0.044). Nine (40.9 per cent) of the instances of dystocia resulted from maternal excitement, mainly attributable to disturbances caused by human beings. In 12 of the cases that resulted in the death of the fawn, the request for veterinary assistance had been delayed at least one day. There were six cases in which the fawns survived; these were the ones that had received assistance within six hours after the onset of parturition.

Laparoscopic repair of renal artery aneurysm.

PURPOSE: We describe technical considerations of the laparoscopic repair of a renal artery aneurysm. MATERIALS AND METHODS: A 57-year-old woman presented with a 3 cm. aneurysm of the distal left main renal artery at its bifurcation. Using a purely laparoscopic 4-port transperitoneal technique the aneurysm was completely mobilized from its location behind the renal vein. Its 3 feeding vessels were controlled individually with bulldog clamps. The aneurysm sac was bivalved and precisely trimmed to conform with the diameter of the main renal artery. Vascular reconstruction was performed with running freehand laparoscopic suturing and intracorporeal knot tying using 4-zero polypropylene suture. RESULTS: Warm ischemia time was 31 minutes, total operative time was 4.2 hours, blood loss was 100 cc and hospital stay was 2 days. Postoperatively renal scan showed improved perfusion and renal arteriography confirmed adequate repair of the aneurysm. CONCLUSIONS: Laparoscopic repair of the renal artery aneurysm is feasible. To our knowledge we present the initial clinical report of laparoscopic renovascular surgery in the literature.

Retroperitoneoscopic adrenalectomy: lateral approach.

Laparoscopy has become the essential surgical approach to the adrenal gland at many institutions, including ours. At the Cleveland Clinic, laparoscopic adrenalectomy for benign and malignant adrenal neoplasms can be performed by either the transperitoneal or the retroperitoneal approach. Herein, we present our technique of lateral retroperitoneal laparoscopic adrenalectomy.

Airway hyperresponsiveness to bronchoconstrictor challenge after wood smoke exposure in guinea pigs.

Prior airway exposure to wood smoke induces an increase in airway responsiveness to subsequent smoke inhalation in guinea pigs (Life Sci. 63: 1513, 1998; 66: 971, 2000). To further characterize this airway hyperreactivity, we investigated and compared the airway responsiveness to bronchoconstrictor challenge before and 30 min after sham air exposure or wood smoke exposure in anesthetized and artificially ventilated guinea pigs. Various doses of substance P (0.8-6.4 microg/kg), capsaicin (0.2-3.2 microg/kg), prostaglandin F2alpha (30-3000 microg/kg), histamine (1-8 microg/kg), or acetylcholine (5-20 microg/kg) were intravenously injected at 2-min intervals in successively increasing doses to obtain the dose required to provoke a 200% increase in baseline total lung resistance (ED200). Wood smoke exposure significantly lowered the ED200 of substance P, capsaicin, and prostaglandin F2alpha whereas sham air exposure failed to do so. Furthermore, wood smoke exposure did not significantly alter the ED200 of histamine or acetylcholine. Pretreatment with phosphoramidon (2 mg/kg), an inhibitor of the neutral endopeptidase (the major degradation enzyme of substance P), before smoke exposure did not significantly affect the smoke-induced reduction in ED200 of substance P. Sectioning both cervical vagi before smoke exposure did not significantly alter the smoke-induced reduction in ED200 of capsaicin or prostaglandin F2alpha. These results suggest that airway exposure to wood smoke acutely produces airway hyperresponsiveness to substance P, capsaicin, and prostaglandin F2alpha, but not to histamine or acetylcholine. Since the combination of phosphoramidon and wood smoke exposure did not result in an additive potentiation of smoke-induced airway hyperresponsiveness to substance P, it is suggested that an inhibition of the degradation enzyme of substance P may contribute to this increase in airway reactivity. Furthermore, vagally-mediated bronchoconstriction does not play a vital role in enhanced airway responsiveness to capsaicin or prostaglandin F2alpha.

Strategies for transplantation of cadaveric kidneys with congenital fusion anomalies.

PURPOSE: The dire shortage of cadaveric kidneys has led to a gradual expansion of donor criteria in the transplant community. The use of kidneys with anatomical fusion anomalies is uncommon and has not been well defined in the literature. We evaluated the surgical strategies and postoperative outcomes of transplanting cadaveric kidneys with congenital fusion anomalies. MATERIALS AND METHODS: Three cadaveric kidneys with congenital fusion anomalies were procured and transplanted between May 1994 and November 1999. None of the 3 donors had any significant urological history. All fusion anomalies were identified during the organ procurement process. RESULTS: Anomalies included 1 L-shaped cross-fused ectopic and 2 horseshoe kidneys. All 3 kidneys were procured en bloc. One horseshoe kidney with a narrow isthmus was split and the 2 kidneys were transplanted into separate recipients, while the other horseshoe kidney was transplanted en bloc into a single recipient. The L-shaped kidney was transplanted en bloc into 1 patient. All transplants were successful with a serum creatinine of 1.1 to 1.9 mg/dl. CONCLUSIONS: To our knowledge we present the initial case of transplantation of an L-shaped kidney. Cadaveric kidneys with congenital fusion anomalies may be transplanted successfully using various individual technical strategies based on the specific renal anatomy. As such, these kidneys may be used to maximize the increasingly inadequate donor pool.

Radiofrequency ablation of the kidney: acute and chronic histology in porcine model.

OBJECTIVES: To our knowledge, the chronic histopathologic effects of radiofrequency ablation (RFA) of renal parenchyma have not been extensively documented. In this study, we report the light and electron microscopic features of renal RFA in acute and chronic porcine models. METHODS: Eleven animals underwent renal RFA of the lower pole kidney bilaterally. RFA was performed laparoscopically in 6 acute animals and percutaneously in 5 chronic animals. Acute animals were killed immediately following surgery. One chronic animal each was killed on postoperative day 3, 7, 14, 30, and 90. Histopathologic evaluation of all renal tissue specimens was carried out with light and electron microscopy. RESULTS: Acutely, the renal radiolesion appeared as a yellowish white, well-circumscribed, necrotic area on gross examination, with evidence of extensive coagulative necrosis and marked inflammation on microscopic examination. From day 1 through 90, light and electron microscopy revealed evidence of progressive, irreversible cell death and necrosis with diminishing inflammatory response in the glomeruli, tubules, and renal interstitium. RFA lesions underwent gradual spontaneous resorption of the necrotic area with ultimate autoamputation. CONCLUSIONS: RFA results in necrosis of the ablated renal parenchyma.

Robotic remote laparoscopic nephrectomy and adrenalectomy: the initial experience.

PURPOSE: We evaluated the feasibility of performing laparoscopic nephrectomy and adrenalectomy exclusively by using robotic telepresent technology from a remote workstation and compared outcomes with those of conventional laparoscopy in an acute porcine model. MATERIALS AND METHODS: Five pigs underwent bilateral laparoscopic nephrectomy (robotic in 5 and conventional in 4) and adrenalectomy (robotic in 4 and conventional in 3). In the 9 robotic laparoscopic procedures all intraoperative manipulations were completely performed telerobotically from a remote workstation without any conventional laparoscopic assistance on site. Animals were sacrificed acutely. RESULTS: Robotic laparoscopic nephrectomy required significantly longer total operative (85.2 versus 38.5 minutes, p = 0.0009) and actual surgical (73.4 versus 27.5 minutes, p = 0.0002) time than conventional laparoscopy. However, blood loss and adequacy of surgical dissection were comparable in the 2 groups. Robotic laparoscopic adrenalectomy required longer total operative (51 versus 32.3 minutes, p = 0.13) and actual surgical (38.5 versus 18.7 minutes, p = 0.14) time than conventional laparoscopy. The solitary complication in this study was an inferior vena caval tear during robotic right adrenalectomy, which was adequately repaired by sutures telerobotically in a remote manner. CONCLUSIONS: To our knowledge we present the initial experience with remote telerobotic laparoscopic nephrectomy and adrenalectomy. Telepresent laparoscopic surgery is feasible.

Effects of aging on mitochondrial enzyme activity of rat urinary bladder.

OBJECTIVES: Our previous study showed that aged rat bladders became fatigued faster than young bladders following repeated contraction induced by electrostimulation. One factor might be a lower energy-producing capability secondary to a decreased mitochondrial enzyme activity of the aged bladder. This study examined this possibility. MATERIALS AND METHODS: Mitochondria from 3- (n = 11) and 24-month-old (n = 10) Sprague-Dawley rats were isolated. Activities of the following enzymes were assayed: two key enzymes in the citric acid cycle, citrate synthase and malate dehydrogenase, and three enzymes in the respiratory chain reaction, NADH-cytochrome c reductase, succinate-cytochrome c reductase and cytochrome c oxidase. The concentration of phosphocreatine and ATP in the aged rat bladders and a separate group of young bladders (n = 12) was determined using high-performance liquid chromatography. RESULTS: (1) The aged bladders have a significantly lower level of phosphocreatine and ATP content than those of young bladders. (2) The activities of all five enzymes assayed were significantly lower in the aged bladders than in young bladders, especially for citrate synthase, which had only 46.8% of the activity of young bladders. CONCLUSIONS: Aging reduces the mitochondrial enzyme activity of the rat bladder resulting in a lower energy-production capability, which might explain some of the voiding dysfunctions found in the elderly.

Interstitial colocalization of two cervid satellite DNAs involved in the genesis of the Indian muntjac karyotype.

A number of repetitive DNA clones were generated from PCR amplifications of Indian muntjac genomic DNA using primer sequences derived from a white tailed deer satellite II DNA sequence. One clone (Mmv-0.7) was characterized and shown to be a cervid satellite II DNA clone. Multiple colored FISH studies with cervid satellite I (C5) and this satellite II clone (Mmv-0.7) to Chinese muntjac metaphase chromosomes localized both satellite DNAs at the pericentromeric regions of all chromosomes except for chromosome 3 and the Y chromosome, whereas chromosome 3 exhibited pericentromeric satellite II DNA only. Where distinguishable, the pericentromeric satellite II signals appeared terminally oriented with respect to satellite I. Six pairs of Chinese muntjac autosomes had interstitial satellite I sites with four of these autosomal pairs (chromosomes 1, 2 and two other smaller autosomal pairs) also exhibiting interstitial satellite II signals. An interstitial site on the X chromosome was found to have satellite II signals. For the Indian muntjac chromosomes, FISH studies revealed a pericentromeric hybridization for satellites I and II as well as 27 distinct interstitial hybridization sites, each having at least one of the satellite DNAs. These data were used to more precisely define the chromosome fusion-associated breakpoints that presumably led to the formation of the present-day Indian muntjac karyotype. It further hints at the possibility that the Indian muntjac karyotype may have evolved directly from a 2n = 70 ancestral karyotype rather than from an intermediate 2n = 46 Chinese muntjac-like karyotype.

Direct visualization of the genomic distribution and organization of two cervid centromeric satellite DNA families.

Several repetitive DNA fragments were generated from PCR amplifications of caribou DNA using primer sequences derived from the white-tailed deer satellite II DNA clone OvDII. Two fragments, designated Rt-0.5 and Rt-0.7, were sequenced and found to have 96% sequence similarity. These caribou clones also had 85% sequence similarity with OvDII. Multiple-colored fluorescence in situ hybridization (FISH) studies with satellite I and satellite II DNA probes to caribou metaphase chromosomes and extended chromatin fibers provided direct visualization of the genomic organization of these two satellite DNA families, with the following findings: (1) Cervid satellite I DNA is confined to the centromeric regions of the acrocentric autosomes, whereas satellite II DNA is found at the centromeric regions of all chromosomes except for the Y. (2) For most acrocentric chromosomes, the satellite I signal appeared to be medially located at the primary constriction, in contrast to that of satellite II, which appeared to be oriented toward the lateral sides as two separate fluorescent dots. (3) The satellite II clone Rt-0.7 appeared to be enriched in the centromeric region of the caribou X chromosome, a pair of biarmed autosomes, and a number of other acrocentric autosomes. (4) Fiber-FISH demonstrated that the satellite I and satellite II arrays were juxtaposed. On highly extended chromatin fibers, the total length of the hybridization signals for the two satellite DNA arrays often reached 300-400 microm. The length of a given satellite II array usually reached 200 microm, corresponding to 2 x 10(3) kb of DNA in a given centromere.

Laparoscopic and percutaneous radiofrequency ablation of the kidney: acute and chronic porcine study.

OBJECTIVES: The chronic effects of renal radiofrequency ablation are unknown. Herein, we investigate the anatomic and physiologic sequelae of laparoscopic and percutaneous renal radiofrequency ablation in acute and chronic porcine models. METHODS: Our study comprised two phases-an acute phase and a chronic phase. In the acute phase, bilateral laparoscopic renal radiofrequency ablation was performed in 6 animals (12 renal units), which were euthanized immediately after surgery. In the chronic study, bilateral percutaneous renal radiofrequency ablation was performed in 5 animals (10 renal units). One animal each was euthanized at postoperative day 3, 7, 14, 30, and 90. RESULTS: Ultrasound-monitored laparoscopic (n = 12) and percutaneous (n = 10) radiofrequency ablations of the lower pole of the kidney were technically successful in each instance. No intraoperative complications occurred. In the survival experiments, the radiolesions showed gradual spontaneous resorption and ultimate renal autoamputation, while maintaining pelvocalyceal integrity as confirmed by ex vivo retrograde ureteropyelogram. Serum creatinine and hematocrit remained stable in all survival animals. Postoperative complication occurred in 1 chronic animal with nonobstructive small bowel dilation at autopsy. CONCLUSIONS: Laparoscopic and percutaneous renal radiofrequency ablation are technically feasible. The anatomic and physiologic sequelae of renal radiosurgery are favorable. Improved techniques of real-time monitoring of the evolving renal radiolesion are necessary.

Laparoscopic aortorenal bypass.

BACKGROUND AND PURPOSE: Open surgical renovascular repair, although producing excellent results, confers significant operative morbidity. As a result, less morbid procedures such as percutaneous balloon angioplasty and stenting have gained increasing acceptance. Laparoscopic techniques have not previously been applied to renal revascularization. The aim of this study was to demonstrate the technical feasibility and the long-term clinical and pathologic outcomes of laparoscopic aorto-left renal artery bypass in a chronic porcine model. MATERIALS AND METHODS: Eight animals were used in the study. All laparoscopic suturing and knot-tying were performed intracorporeally using free-hand techniques. The following operative steps were employed: (1) aortic dissection and cross-clamping; (2) transection and refashioning of the left renal artery ostium; (3) in-situ renal hypothermia; (4) end-to-side aorto-left renal artery anastomosis; and (5) aortic unclamping. In situ renal hypothermia was achieved laparoscopically by infusion of ice-cold heparinized saline into the renal artery through a balloon catheter. RESULTS: All eight pigs underwent laparoscopic aortorenal bypass successfully. The median surgical time was 110 minutes, and the aortic cross-clamping time was 45.5 minutes. The median anastomotic time was 40 minutes, and the renal warm ischemia time was 9 minutes. The median estimated blood loss was 30 mL. An intraoperative complication of suture breakage leading to anastomotic hemorrhage occurred in one animal; the problem was corrected laparoscopically. Postoperatively, one animal died from pneumonia. The remaining seven animals experienced no postoperative complications and were euthanized, one each at postoperative day 0 and 1 and week 1, 2, 3, 4, and 6. The median preoperative and postoperative (at euthanasia) serum creatinine values (1.15 mg/dL v 1.2 mg/dL; P = 0.39) were similar. However, compared with preoperative peripheral renin activity (0.25 microg/L per hour), the postoperative peripheral renin activity was elevated (0.9 microg/L per hour; P = 0.047). Autopsy revealed a grossly normal left kidney, with Doppler confirmation of flow in the repaired renal artery in all seven animals. Ex vivo angiography confirmed a patent anastomosis. On histopathology examination, the early left renal parenchymal specimens revealed transient, mild acute tubular necrosis that resolved over sequential specimens without significant long-term sequelae. Histologic analysis of the aorto-left renal artery anastomotic site revealed gradual endothelialization with time. CONCLUSIONS: Laparoscopic aortorenal artery revascularization is technically feasible. Our chronic animal model has demonstrated durable success over a 6-week follow-up. This study represents the initial report in the literature.