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T Bukowski

Publications and source records attributed to T Bukowski.

9 recordsLinked to original sources

Precise, highly female-biased sex ratios in a social spider.

It has been recognized for some time that the risk of producing maleless clutches should select for a lower than binomial variance in the sex ratio of organisms with female-biased sex ratios, small clutches and breeding groups containing the clutch of a single female. However, to date, precise sex ratios have only been reported for organisms with haplodiploid sex determination, a system which allows direct control of the sex of individual offspring. In contrast, under heterogametic sex determination chance is expected to play a crucial role in determining the sex composition of any one family, in particular when males are the heterogametic sex. Here, we present evidence of precise or underdispersed primary sex ratios in the Neotropical social spider Anelosimus domingo Levi. We show that this diplodiploid species with male heterogamety has not only beaten the odds of meiosis by producing mostly daughters, but has also attained relative precision in the proportion of sons and daughters produced in any one clutch. The latter finding suggests the existence of mechanisms that allow sorting of the two types of sperm in this spider species.

Animals↗

Development of the methylotrophic yeast Pichia methanolica for the expression of the 65 kilodalton isoform of human glutamate decarboxylase.

We describe a protein expression system in the methylotrophic yeast, Pichia methanolica. Methods for transformation and genetic manipulation of the organism were developed using an ade2 strain and the wild-type ADE2 gene. A vacuolar protease-deficient strain was constructed. Two genes encoding alcohol oxidases were found, yet a single isoform of alcohol oxidase was produced during methanol-fed fermentations. The promoter from this gene was used to drive expression. An integrating plasmid for the cytoplasmic expression of the 65 kDa isoform of human glutamate decarboxylase (human GAD65) was assembled. A strain harboring eight copies of this plasmid expressed enzymatically active human GAD65 at levels approaching 0.5 g/l. Identical amounts were made in Pichia pastoris. The recombinant GAD65 was purified to greater than 90% purity.

Alcohol Oxidoreductases↗

New application of the gastrostomy button for clinical and urodynamic evaluation before vesicostomy closure.

PURPOSE: We report use of the Bard gastrostomy button to occlude vesicostomy and provide access for intermittent catheterization before closure in children with vesicostomy. Evaluation of bladder function in such children usually relies on radiographic and urodynamic studies, which may fail to predict bladder compliance, emptying and continence after closure. MATERIALS AND METHODS: Buttons were placed before vesicostomy closure in 1 boy with the prune-belly syndrome and 2 girls with cloacal anomalies 2.5 to 10 years old. The patients had undergone vesicostomy using the Blocksom technique soon after birth because of urinary infection, and impairment of bladder emptying and renal function. RESULTS: With the button in place bladder emptying, compliance, continence and possible upper tract changes could be evaluated during several weeks. Button coaptation to the vesicostomy was complete with no urine leakage around the device, allowing easy intermittent drainage through its channel. After 4 weeks the buttons did not have any encrustation or lithiasis and patients were free of urinary infection. Vesicostomy was closed in each patient and the period of temporary closure provided by the button was predictive of future bladder behavior. CONCLUSIONS: This new and original application of the gastrostomy button as a temporary vesicostomy closure may be useful to predict the clinical and urodynamic responses of a defunctionalized bladder in patients with vesicostomy who are candidates for urinary undiversion.

Child↗

Is there a best alternative to treating the obstructed upper pole?

PURPOSE: We addressed whether salvage of upper pole renal units in comparison to partial nephrectomy affects differential renal function in patients with a duplicated obstructed upper pole. MATERIALS AND METHODS: We retrospectively reviewed the records of all children who underwent surgery for a unilateral obstructed duplicated kidney at our institution from 1988 to 1995. Patients were evaluated with respect to postoperative complications, reoperation rate and percent change in differential renal function of the obstructed duplicated kidney, as determined by nuclear renography. RESULTS: We identified 46 patients with a unilateral obstructed upper pole of a duplicated kidney who were divided into 2 groups. Group 1 (12 patients) underwent an upper pole salvage procedure, that is ureteropyelostomy or ureteroureterostomy, and group 2 (31 patients) underwent partial nephrectomy. Three reoperations (25%) were performed in group 1 and 1 (4%) was done in group 2. Postoperative symptomatic urinary tract infections were diagnosed in 3 group 1 patients (25%) and in 2 (8%) in group 2. Average change in ipsilateral renal function in the 8 patients who underwent upper pole salvage procedures was 2.25 +/- 2.34% (range -6 to 12). In the 8 patients who underwent upper pole nephrectomy and who also had postoperative renal scans average change in function was -1.25 +/- 4.51% (range -23 to +16). CONCLUSIONS: There was no statistically significant loss of relative renal function in patients treated with partial nephrectomy and no significant gain in relative renal function in those treated with an upper pole salvage procedure. The reoperation rate was higher in the upper pole salvage than in the partial nephrectomy group (25 versus 4%). While not statistically significant, we believe that this rate is clinically important. We think that partial nephrectomy should remain the preferred treatment for most patients with obstructed duplicated kidneys.

Adolescent↗

The use of metrizamide for stabilizing against convection in sedimentation equilibrium.

In meniscus depletion sedimentation equilibrium experiments, the solute concentration gradient in much of the cell is insufficient to stabilize against thermal convection. Experiments with spectrin at 35 degrees C led to sufficient convection that, particularly at low concentrations of solute, no apparent redistribution occurred. At low angular velocity, the density gradient provided by moderate concentrations of NaCl or additional sucrose is inadequate to overcome the convective disturbances. We have found that the use of low concentrations of metrizamide can overcome these problems. The higher molecular weight and lower partial specific volume of metrizamide vis-a-vis sucrose allow a significant and stabilizing density gradient even at angular velocities below 8000 rpm.

Centrifugation, Density Gradient↗

Self-association and active enzyme forms of Naja naja naja and Crotalus atrox phospholipase A2 studied by analytical ultracentrifugation.

The dimerization of phospholipase A2 (PLPA2) from Naja naja naja (Pakistani cobra) and Crotalus atrox (Western Diamondback rattlesnake) has been studied from pH 2.5 to 11 at 20 degrees C in 1 mM CaCl2, 0.21 M ionic strength. For the C. atrox enzyme, it was found necessary to use a combination of sedimentation equilibrium and fluorescence yield data to analyze the association. Sedimentation equilibrium in the analytical ultracentrifuge sufficed for the study of the N. naja PLPA2. In the region of enzymatic activity at pH 8, the dimerization association constants found were k2 = 2.8 X 10(6) L/mol and k2 = 6.9 X 10(4) L/mol for the C. atrox and N. naja enzymes, respectively. Analytical linked functions are presented which describe the data. Because the associations are linked to Ca2+ as well as the hydrogen ion, no attempt was made to interpret the ionization of residues in terms of the molecular structure. Active-enzyme sedimentation velocity experiments have been used to study the relation between enzymatic activity and association for both the C. atrox and N. naja enzymes. The substrate 1,2-dibutyryl-sn-glycero-3-phosphocholine (diC4PC) did not dissociate the C. atrox PLPA2. The substrate 1,2-dihexanoyl-sn-glycero-3-phosphocholine (diC6PC) at 7.5 mM dissociated the C. atrox PLPA2 when monitored either as the active enzyme or as the Sr2+-inhibited enzyme. At low enzyme concentrations, 40 mM diC4PC had no effect on N. naja PLPA2 dimerization. However, the sedimentation coefficients observed at enzyme concentrations above 0.2 mg/mL in active-enzyme sedimentation velocity experiments were larger than the values predicted from the thermodynamic studies. Sedimentation coefficients observed for the N. naja PLPA2 acting on diC6PC were larger than those of the monomeric protein, which was the form layered on this substrate. The dissociation of the C. atrox PLPA2 effected by diC6PC was analyzed by the thermodynamics of association and the kinetic Michaelis constant. The analysis suffices to account for the observed sedimentation coefficient. The sedimentation behavior of the N. naja PLPA2 acting on diC6PC substrate was analyzed in terms of a protein-lipid complex. With this model, 68 +/- 16 phospholipid molecules per protein monomer were determined. It is proposed that this enzyme has two micelle nucleation sites per monomer. These putative sites promote micelle formation of the substrate on the enzyme below the critical micelle concentration of the lipid alone.

Animals↗

Integral.

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Hospital Departments↗