Expression and characterization of an extremely thermostable ß-glycosidase (mannosidase) from the hyperthermophilic archaeon Pyrococcus furiosus DSM3638.

Genomic analysis of the hyperthermophilic archaeon Pyrococcus furiosus revealed the presence of an open reading frame (ORF PF0356) similar to the enzymes in glycoside hydrolase family 1. This ß-glycosidase, designated PFTG (P. furiosus thermostable glycosidase), was cloned and expressed in Escherichia coli. The expressed enzyme was purified by heat treatment and Ni-NTA affinity chromatography. The gene was composed of 1,452 bp encoding 483 amino acids for a protein with a predicted molecular mass of 56,326 Da. The temperature and pH optima were 100°C and 5.0 in sodium citrate buffer, respectively. The substrate specificity of PFTG suggests that it possesses characteristics of both ß-galactosidase and ß-mannosidase activities. However, through kinetic studies by ITC (Isothermal Titration Colorimetry) which is very sensitive method for enzyme kinetics, PF0356 enzyme revealed the highest catalytic efficiency toward p-nitrophenyl-ß-d-mannopyranoside (3.02 k(cat)/K(m)) and mannobiose (4.32 k(cat)/K(m)). The enzyme showed transglycosylation and transgalactosylation activities toward cellobiose, lactose and mannooligosaccharides that could produce GOS (galactooligosaccharides) and MOS (maltooligosaccharides). This novel hyperthermostable ß-glycosidase may be useful for food and pharmaceutical applications.

Perineal lipoma in association with scrotal anomalies in children.

OBJECTIVE: To analyse the characteristics of the perineal lipoma and speculate on its development, as an associated perineal lipoma is implicated in the occurrence of scrotal anomalies in children. PATIENTS AND METHODS: Five children with a perineal lipoma in association with various scrotal anomalies were included in the study. We analysed the types of scrotal anomalies and their anatomical relationship with the perineal lipomas. RESULTS: One child had a supra-inguinal ectopic scrotum, and the others had penoscrotal transposition, either symmetrical or asymmetrical; they included two who had an accessory scrotum combined with a perineal lipoma. The perineal lipoma was on the ipsilateral side of the upward-positioned hemiscrotum in the ectopic scrotum or the asymmetrical penoscrotal transposition, or it was on the mid-perineum in the symmetrical penoscrotal transposition. CONCLUSIONS: The perineal lipoma can be found combined with various scrotal anomalies, including ectopic scrotum as well as an accessory scrotum or penoscrotal transposition; this is the first report describing a perineal lipoma associated with an ectopic scrotum. We speculate that the development of the perineal lipoma is closely related to scrotal anomalies, which result from early division and/or abnormal migration of the labioscrotal swelling. The concomitant perineal lipoma might be formed during the process, and might affect abnormal scrotal development.

Enuresis alarm treatment as a second line to pharmacotherapy in children with monosymptomatic nocturnal enuresis.

PURPOSE: We evaluate the effectiveness of enuresis alarm as second line therapy for partial or nonresponders to pharmacotherapy. MATERIALS AND METHODS: We recommended enuresis alarm treatment for 67 partial or nonresponders to pharmacotherapy for monosymptomatic nocturnal enuresis. Of these patients 28 were evaluable by inclusion and exclusion criteria. We analyzed the effectiveness of enuresis alarm therapy using the response criteria defined by the reduction rate of wet nights, defined as complete response (greater than 90%), partial response (50% to 90%) and no response (less than 50%). We defined initial and lasting cure when patients showed persistent full response for 4 weeks and 6 months after cessation of all treatments, respectively. RESULTS: After pharmacotherapy partial responders showed a mean response of 81.2%. On the other hand, nonresponders exhibited a mean response of 26.4% with more than 20 wet nights in 4 weeks. After using second line enuresis alarm treatment 90.5% (19 of 21) of partial responders became full responders and 71.4% (15 of 21), 61.9% (13 of 21) of partial responders showed initial and lasting cure, respectively. In addition, 71.4% (5 of 7) of nonresponders became full responders and 57.1% (4 of 7) exhibited initial and lasting cure. CONCLUSIONS: Enuresis alarm as second line therapy for monosymptomatic nocturnal enuresis is effective for lasting cure as well as high initial full response rate in partial and nonresponders to pharmacotherapy. Therefore, enuresis alarm is a reasonable second line therapeutic option for partial or nonresponders to pharmacotherapy.

Serial followup of the contralateral renal size in children with multicystic dysplastic kidney.

PURPOSE: Multicystic dysplastic kidneys have negligible renal function and the contralateral kidney (solitary kidney) frequently exhibits abnormalities that may affect growth. We previously showed that nomograms related to renal size constructed from digitalized ultrasonographic measurements of renal parenchymal area are a sensitive measure of renal growth and correlate with functional mass. We assess the age-dependent characteristics of compensatory renal growth in infants and children with multicystic dysplastic kidneys by construction of a growth curve for the contralateral kidney, assess these characteristics in comparison to normal renal growth of right and left kidneys, analyze the extent of compensatory renal growth and evaluate abnormal growth in solitary kidneys in infants and children. MATERIALS AND METHODS: From 1988 to 2000 we reviewed 152 serial sonograms from 48 patients with a diagnosis of multicystic dysplastic kidneys. We also reviewed 209 renal sonograms in patients whose studies, done for other purposes, showed normal bilateral kidneys. Using computer planimetry, parenchymal area and pelvicaliceal area were determined after digitalization of ultrasound images. Parenchymal area was calculated by parenchymal area minus pelvicaliceal area and expressed as a mean of 3 measurements. A parenchymal area growth curve was generated for the contralateral kidney in the multicystic dysplastic kidney group from birth to 216 months, and for right and left normal kidneys from birth to 338 months. Data were plotted as mean parenchymal area +/- 2 SD on a nomogram generated by linear regression. Differences in parenchymal area between normal right and left kidneys, between normal kidneys and the contralateral to multicystic dysplastic kidney were analyzed by unpaired Student t test. RESULTS: Of the 48 patients with multicystic dysplastic kidneys 36 had contralateral normal kidneys and 12 (25%) had a contralateral abnormality. Of the 12 cases 4 and an additional 5 without an identified abnormality (9 of 48) or 18.7% had solitary kidneys 2 SD below the normal growth curve for total parenchymal area, indicating a smaller than expected increase in compensatory renal growth. Conversely, 8 of 12 including 1 with grade V reflux into a solitary kidney exhibited normative compensatory renal growth. Left normal kidneys demonstrated a small but statistically significantly larger parenchymal area throughout growth. Solitary kidneys did not demonstrate growth differences associated with side. Solitary kidneys showed accelerated growth from 0 to 22 months while normal kidneys showed accelerated growth from 0 to 15 months. CONCLUSIONS: Nomograms constructed from ultrasonographic measurements of renal parenchymal area may be useful for assessing abnormal renal growth in solitary kidneys. Patients with solitary kidneys identified by conventional ultrasonographic measurement as normal may not exhibit expected growth. Clinical decision making may be improved by identification of solitary kidneys at risk for poor growth.