Isolation and characterization of a novel 5-oxoprolinase (without ATP-hydrolyzing) from Alcaligenes faecalis N-38A

A screening test was undertaken to isolate a microorganism that produced 5-oxoprolinase (without ATP-hydrolyzing). The 5-oxoprolinase (without ATP-hydrolyzing) activity (decyclization activity toward L-pyroglutamate) was found in a cell-free extract of Alcaligenes faecalis N-38A, newly isolated from a soil sample. The enzyme was purified as a homogeneous preparation. The molecular weight of the enzyme was estimated to be 47,000. The decyclization activity was specific for L-pyroglutamate, and independent of ATP and metal ions. The reaction was a reversible one, i.e., cyclization reaction of L-glutamate to yield pyroglutamate was identified.     

Leukoregulin induction of protein expression in human orbital fibroblasts: evidence for anatomical site-restricted cytokine-target cell interactions

The molecular basis for the profound inflammatory response and the accumulation of hyaluronan in orbital connective tissues seen in thyroid-associated ophthalmopathy is unknown. Moreover, the link between the orbital manifestations of Graves' disease and those in the pretibial skin, localized dermopathy, has yet to be established. We have reported recently that leukoregulin, an activated T lymphocyte-derived cytokine, dramatically induces hyaluronan synthesis and prostaglandin-endoperoxide H synthase 2 in human orbital fibroblasts in culture. In the current studies, utilizing giant two-dimensional gel electrophoresis, we find that orbital fibroblasts express constitutively a protein profile that distinguishes them from skin fibroblasts derived from the abdominal wall and from the pretibium. We further demonstrate that leukoregulin, when present in culture medium for 16 hr, up-regulates a set of orbital fibroblast proteins not present in untreated cultures or in fibroblasts from the abdominal wall. However, some of the same protein inductions are present in the pretibial fibroblasts. These leukoregulin-induced changes in protein expression are completely blocked by dexamethasone (10 nM). Our findings are the first to identify proteins that appear to be expressed and differentially regulated in an anatomical site-restricted manner in orbital and pretibial fibroblasts and seem to establish a molecular link between fibroblasts from the orbit and those in pretibial skin.     

Clonal composition of malignant fibrous histiocytoma: analysis by PCR-based assay of the human androgen receptor gene (HUMARA)

Poly(A)+ mRNA was isolated from leaves of potato plants (Solanum tuberosum L. cv. Desiree) according to standard protocols. This poly(A)+ mRNA was injected via glass microcapillaries into oocytes that were surgically removed from the African clawed toad Xenopus laevis. As a control, oocytes were either injected with H2O or remained untreated. Three days after injection the oocytes were analyzed by two electrode voltage clamping. Current voltage analysis revealed that a K+ channel from potato was functionally expressed in injected oocytes. The identity of this K+ channel was confirmed by its substrate specificity and a shift in the reversal potential. In particular, when the outside K+ concentration was increased the reversal potential of poly(A)+ injected oocytes shifted to more positive values. Furthermore, K+ outward currents declined when the outside K+ concentration was raised from 0.1 to 100 mM. Inward currents increased with an elevation of the K+ concentration. Several pharmaceuticals were tested for their potential to block this K+ channel. As a result, the channel was completely blocked by BaCl2. A three state reaction kinetic model was used to simulate the currents through the K+ transport protein as function of the extracellular K+ concentration. In particular, the simulation revealed current voltage relations that exactly matched the measured ones. Saturation of current voltage curves emerged from the simulation as a consequence of high extracellular potassium concentration.     

Klippel-Trenaunay-Weber syndrome with adrenal pseudocyst: characterization by blood pool and adrenocortical iodocholesterol scintigraphy

A 44-year-old woman with Klippel-Trenaunay-Weber syndrome (KTWS) was evaluated for an adrenal mass incidentally discovered on CT. An NP-59 scintigraph showed normal symmetric adrenal tracer uptake, and a Tc-99m labeled RBC scan showed no evidence for an adrenal hemangioma but did show characteristic dilated and tortuous vasculature at multiple other sites. The mass was removed and found to be an adrenal pseudocyst. The pseudocyst was believed to have arisen from a pre-existing, thrombosed and organized adrenal hemangioma. This case demonstrates the previously reported utility of RBC scintigraphy in the depiction of the vascular abnormalities in the diagnosis and treatment of patients with Klippel-Trenaunay-Weber syndrome, as well as the utility of NP-59 scintigraphy in evaluating patients with adrenal masses.     

Long-term results of total penile reconstruction with a prefabricated lateral arm free flap

Trifluoroethanol (TFE) is often used to increase the helicity of peptides to make them usable as models of helices in proteins. We have measured helix propensities for all 20 amino acids in water and two concentrations of trifluoroethanol, 15 and 40% (v/v) using, as a model system, a peptide derived from the sequence of the alpha-helix of ribonuclease T1. There are three main conclusions from our studies. (1) TFE alters electrostatic interactions in the ribonuclease T1 helical peptide such that the dependence of the helical content on pH is lost in 40% TFE. (2) Helix propensities measured in 15% TFE correlate well with propensities measured in water, however, the correlation with propensities measured in 40% TFE is significantly worse. (3) Propensities measured in alanine-based peptides and the ribonuclease T1 peptide in TFE show very poor agreement, revealing that TFE greatly increases the effect of sequence context.     

Phase I trial of weekly scheduling and pharmacokinetics of titanocene dichloride in patients with advanced cancer

PURPOSE: To determine the maximum-tolerated dose (MTD) and the dose-limiting toxicities (DLTs) of a weekly schedule of titanocene dichloride (TD) and to define the pharmacokinetics of titanium in plasma and urine. PATIENTS AND METHODS: Twenty patients with a median age of 58 years received 83 courses of TD. TD was given as 1-hour infusion at escalating doses from 70 to 185 mg/m2/wk. Pharmacokinetic analysis was performed in eight patients for total plasma titanium (TPTi) and in three patients for ultrafiltrable titanium (UFTi). RESULTS: At the fifth dose level (185 mg/m2/wk), a variety of DLTs were seen in five patients: fatigue in three, bilirubinemia in one, and hypokalemia in two. A further six patients were treated at 140 mg/m2; only one had dose-limiting creatinine elevation and this dose was therefore defined as the MTD. No myelosuppression or alopecia were observed. One patient with adenocarcinoma of unknown primary had a minor response. Pharmacokinetic analysis showed that TPTi maximum concentration (Cmax) values were linear with dose and elimination of TPTi was triphasic with a long terminal half-life (t1/2; median, 165 hours; range, 89 to 592). Between 7% and 24.3% of the total of administered titanium was eliminated in urine over the first 24 hours. In contrast, UFTi elimination was described by a one-compartment model with a t1/2 of 0.41 hours; peak levels of UFTi were 5.2% +/- 2.5% those of TPTi. CONCLUSION: The MTD of TD given on a weekly schedule is 140 mg/m2, with cumulative, but reversible creatinine and bilirubin elevation being the DLTs.     

Temperature monitoring

Central temperature is usually tightly regulated in human beings. Anesthesia alters the normal thermoregulatory controls of the body. Intraoperatively, mild degrees of hypothermia may provide some cerebral protection. However, the risk of organ dysfunction and shivering require that the anesthesiologist be prepared to treat severe hypothermia. Appropriate measures such as warning the operating room and using forced air blankets can prevent both intraoperative hypothermia and postoperative shivering. The use of temperature measurement is not limited to the operative and immediate recovery periods. Anesthesiologists practicing in intensive care units and in pain clinics use temperature monitoring as a diagnostic tool in a variety of situations.     

A note on circle packing

The problem of packing circles into a domain of prescribed topology is considered. The circles need not have equal radii. The Collins-Stephenson algorithm computes such a circle packing. This algorithm is parallelized in two different ways and its performance is reported for a triangular, planar domain test case. The implementation uses the highly parallel graphics processing unit (GPU) on commodity hardware. The speedups so achieved are discussed based on a number of experiments.     

Determination of porous glass surface free energy components from contact angles

The contact angles of water drops and diiodomethane drops on pellets made of controlled porosity glasses have been measured. The surface of the glasses was modified by thermal treatment at 873 K which led to an increase in the surface concentration of boron atoms. Glass modified with Carbowax 20M (polyethylene glycol) and fully hydroxylated glass have also been studied. Using the measured contact angles and modified Young equation, the dispersion and nondispersion components of the glass surface free energy have been calculated. The values show that with increasing heating time (increasing surface density of boron atoms) an increase in the surface polarity takes place. However, the polarity of the same samples decreases after treatment with Carbowax, increasingly so with higher boron atom concentrations.