Intracellular retention and degradation of tissue-nonspecific alkaline phosphatase with a Gly317-->Asp substitution associated with lethal hypophosphatasia

One point mutation which converts glycine-317 to aspartate of tissue-nonspecific alkaline phosphatase (TNSALP) was reported to be associated with lethal hypophosphatasia (Greenberg, C. R., et al. Genomics 17, 215-217, 1993). In order to define the molecular defect of TNSALP underlying the pathogenesis of hypophosphatasia, we have examined the biosynthesis of TNSALP with a Gly317-->Asp substitution. When expressed in COS-1 cells, the mutant did not exhibit alkaline phosphatase activity at all, indicating that the replacement of glycine-317 with aspartate abolishes the catalytic activity of TNSALP. Pulse-chase experiments showed that the newly synthesized mutant failed to acquire Endo H-resistance and to reach the cell surface. Interestingly, this TNSALP mutant was found to form a disulfide-bonded high-molecular-mass aggregate and was rapidly degraded within the cell, though the mutant protein was modified by glycosylphosphatidylinositol (GPI). Lactacystin, an inhibitor of the proteasome, obstructed the degradation of the mutant protein, suggesting the involvement of proteasome as a part of quality control of TNSALP.     

Design, synthesis and evaluation of transition-state analogue inhibitors of Escherichia coli gamma-glutamylcysteine synthetase

Phosphinic acid-, sulfoximine- and sulfone-based transition-state analogues were synthesized and evaluated as inhibitors of Escherichia coli gamma-glutamylcysteine synthetase. These compounds have a carboxyl function at the beta-carbon to the tetrahedral central hetero atom so as to mimic the carboxyl group of the attacking cysteine in the transition state. The phosphinic acid- and the sulfoximine-based compounds were found to be potent ATP-dependent inactivators, both showing a slow-binding kinetics with overall affinities and second-order inactivation rates of one to two orders of magnitude greater than those of L-buthionine (SR)-sulfoximine (L-BSO). The sulfone was a simple reversible inhibitor without causing ATP-dependent enzyme inactivation, but its affinity toward the enzyme was still five times greater than that of L-BSO, indicating that the beta-carboxyl function plays a key role in the recognition of the inhibitors by the enzyme. The sulfoximine with (S)-beta-carbon to the sulfur was synthesized stereoselectively, and the two diastereomers with respect to the chiral sulfur atom were separated as a cyclic sulfoximine derivative. The sulfoximine with R-configuration around the sulfur served as an extremely powerful ATP-dependent inactivator with an overall inhibition constant of 39 nM and an inactivation rate of 6750 M-1 s-1, which correspond to 1260-fold higher affinity and almost 1400-fold greater inactivation rate as compared with L-BSO. The sulfoximine with (S)-sulfur was a simple reversible inhibitor with an inhibition potency comparable to that of the sulfone. The synthesis and inhibition profile of the N-phosphoryl sulfoximine is also described.     

The responses of Verreaux''s sifakas to anti-predator alarm calls given by sympatric ring-tailed lemurs

Erectile dysfunction has an incidence of 2-9% and it is seen often in general practice. But the more recent treatment methods such as intracavernous drug injections and vacuum constriction devices are not known by general practitioners and normally used drug treatment has no efficacy. The management of impotence of the general practitioner should concentrate upon internistic conditions like diabetes, hypertension, hypercholesteremia and different drugs causing impotence. The symptomatic treatment of erectile dysfunction has to be done by a specialist, who is able to offer all therapeutic options.     

Development of a deep-hole, laser-guide boring tool: the boring of workpieces with a thin wall and an inclined prebored hole

A laser-guided BTA tool has been developed to prevent axial hole deviations. Four kinds of experiments were conducted to examine the performance of this tool in detail and to determine its practical application using a duralumin (A2017-T4) workpiece with a prebored 108-mm diameter hole. The first experiment was performed to examine the conditions of the hole deviation in the case of normal deep-hole boring. The second was to examine whether the tool corrects for the stated hole deviation. The third was to examine whether the tool can go straight through to the target without shifting toward a thin wall (t = 0.7 mm) on one side of the workpiece. The fourth was to examine whether the tool can be guided toward the target without affecting a prebored hole, which was inclined to radial direction at 100 μm for a hole length of 100 mm. The experiments were performed with a rotating tool-stationary workpiece system. Rotational speed was 270 rpm and feed ws 0.125 mm/rev. Tool diameter was 110 mm. The results show that the tool can be guided to go straight through toward the target despite the disturbances.     

Spinal meningeal melanocytoma presenting with superficial siderosis of the central nervous system. Case report and review of the literature

1. We recently demonstrated that intrathecal administration of prostaglandin E2 (PGE2) and PGF2alpha induced allodynia through a pathway that includes the glutamate receptor and nitric oxide (NO)-generating systems from pharmacological studies. In order to clarify the involvement of NO in prostaglandin-induced allodynia, we measured NO released from rat spinal cord slices by a chemiluminescence method. 2. PGE2 stimulated NO release from both dorsal and ventral regions all along the spinal cord. PGE2 stimulated the release within 10 min and increased it in a time-dependent manner. 3. The PGE2-induced NO release was observed at 100 nM-10 microM. PGF2alpha stimulated the release at concentrations higher than 1 microM, but PGD2 (up to 10 microM) did not enhance it. 4. 17-Phenyl-omega-trinor PGE2 (EP1 > EP3) and sulprostone (EP1 < EP3) were as potent as PGE2, but PGE1 was less potent, in stimulating NO release. While M&B 28767 (EP3) did not enhance the release, butaprost (EP2) stimulated it at 1 microM. The PGE2-evoked release was blocked by ONO-NT-012, a bifunctional EP1 antagonist/EP3 agonist. 5. The PGE2-evoked release was Ca2+-dependent and blocked by MK-801 (NMDA receptor antagonist) and L-NAME (NO synthase inhibitor). The release was also inhibited by PGD2 and dibutyryl-cyclic AMP. 6. The present study demonstrated that PGE2 stimulates NO release in the rat spinal cord by activation of NMDA receptors through the EP1 receptor, and supports our previous findings that the NO-generating system is involved in the PGE2-induced allodynia.     

A 100-channel optical FDM transmission/distribution at 622 Mb/sover 50 km

A 100-channel optical frequency-division multiplexing (FDM) transmission/distribution experiment at 622 Mb/s is demonstrated for a fiber length of 50 km. The feasibility of a polarization-insensitive waveguide frequency selection switch for 10-GHz intervals and a frequency-shift-keying (FSK) direct-detection scheme employing a Mach-Zehnder filter is verified. The demodulation circuit employs a Mach-Zehnder filter and a balanced receiver, which utilizes optical power more efficiently than the Fabry-Perot filter. No receiver sensitivity degradation is observed due to interchannel crosstalk of the 128-channel tunable waveguide frequency selection switch (FS-SW) or fiber four-wave mixing for transmissions over a 50-km-long nondispersion-shifted (NDS) fiber and a 26-km-long dispersion-shifted (DS) fiber     

Postprostatectomy dysuria: restored voiding function by perineal hot water spray in an old male with a contractile bladder]

This study reports the first case of a subtrochanteric stress fracture of the femur after total knee arthroplasty. A 61-year-old obese woman was treated by right total knee arthroplasty for osteoarthrosis. Four months after the surgery and 1 month after full weight bearing, she complained of tenderness in the right groin and proximal thigh. There was no history of trauma. Radiographs of the right hip and of the right proximal femur were interpreted as unremarkable, and nonsteroid anti-inflammatory drugs were administered. Three months later, she had a sudden onset of increased pain with instability and giving way of the right leg. Radiographs revealed a subtrochanteric transverse fracture of the right femur. Histologic workup of the bone and tissue specimen taken at open reduction, as well as the laboratory data were consistent with the radiologic and clinical diagnosis of a stress fracture. It is most likely that a decrease in the tension band effect of the iliotibial tract in combination with coxa vara and changes in static and dynamic forces of the femur and an increased level of activity after a period of relative inactivity secondary to the knee arthroplasty are responsible for this very uncommon fracture type.     

Multichannel frequency conversion experiment using fibre four-wave mixing

Multichannel frequency conversion is demonstrated using four-wave mixing. A polarisation independent configuration is employed, by which polarisation control for each signal is not necessary. By setting the pump light frequency at the zero-dispersion wavelength of the fibre, three FSK modulated signals with a frequency spacing of 70 GHz are simultaneously converted with an equal efficiency of -27.5 dB. Bit error rate measurements confirm the feasibility of this conversion scheme.<>     

Induction of Ets-1 in endothelial cells during reendothelialization after denuding injury

The requirement of Y-chromosome activity for the differentiation of somatic cells and germ cells was studied in the fetal gonads of X/XSxra mouse embryos where the activity of the Sxra fragment of the Y chromosome is influenced by the inactivation and reactivation of the X chromosome. In the interstitial somatic cells, random inactivation of the X and the XSxra chromosomes took place which was revealed by the mosaic expression of an X-linked lacZ transgene. The Sertoli cells, however, displayed a preferentially active XSxra chromosome and the presence of Sxra-active Sertoli cells was associated with the morphogenesis of testicular tubules in the sex-reversed gonads. The activity of the Y-chromosome fragment is therefore necessary for the differentiation of the Sertoli cells which may direct the development of the testis. The expression pattern of the X-linked transgene in X/XSxra germ cells suggests that both the X and the XSxra chromosomes are active. This finding suggests that the presence of Sxra has no impact on the reactivation of the X chromosome in the germ cells and that the X chromosome can be reactivated even though the germ cells are found in the testicular environment. Our results are consistent with the concept that the activity of genes on the XSxra fragment is essential for the differentiation of Sertoli cells and the morphogenesis of the testis, but not for premeiotic differentiation of germ cells in sex-reversed mice.