Purification and some properties of ribonuclease from Xenopus laevis eggs

A 122 kDa RNase from eggs of Xenopus laevis was purified by sequential chromatography on Sephadex G-75, DEAE-cellulose, heparin-Sepharose and TSK gel G3000SW columns, and gave a single 60 kDa band on SDS-polyacrylamide gel electrophoresis under reducing and nonreducing conditions. The RNase composed of two 60 kDa subunits is able to recognize pyrimidine bases specifically. The pH optimum of the RNase was 7.5 in Tris-HCl buffer. The enzyme activity was abolished by treatment at 80 degrees C for 5 min and pH 2 or 12 for 1 h. Since egg lectins with RNase activity obtained from Rana catesbeiana and R. japonica and bovine pancreatic RNase A show about 30% protein homology and these three proteins are 12-14 kDa heat-stable RNases, [K. Titani, K. Takio, M. Kuwada, K. Nitta, F. Sakakibara, H. Kawauchi, G. Takayanagi and S. Hakomori, Biochemistry, 26, 2189 (1987); Y: Kamiya, F. Oyama, R. Oyama, F. Sakakibara, K. Nitta, H. Kawauchi, Y. Takayanagi and K. Titani, J. Biochem. (Tokyo), 108, 139 (1990)], the data suggest that the X. laevis egg RNase is a unique protein compared with RNases from not only amphibians, but also mammals.     

Morphogenesis of degenerative changes predisposing dogs to rupture of the cranial cruciate ligament

The cranial cruciate ligaments (CCL) of 13 dogs with clinical signs of CCL rupture and those of 22 clinically healthy young beagle dogs for laboratory use were examined histopathologically and immunohistopathologically. The most constant changes at an early stage of degenerating ligament tissue in affected dogs were nuclear enlargement and perinuclear halo formation of fibrocytes followed by chondroid metaplasia. These changes were also frequent in apparently healthy young beagles kept under laboratory conditions. PAS and alcian blue positive substance accumulated around activated fibrocytes and within perinuclear halos. S-100 protein was also positive in these cells preceding the morphological change of chondroid metaplasia. Increased mitotic figures and Ki-67 positive cells showed the proliferating nature of these cells at a later stage. Alteration of extracellular matrices from dense collagen fiber type to those of cartilage tissue seemed to predispose dogs to rupture of the CCL along with a degradation in collagen fiber of the primary bundles. Collagen fiber bundles with a parallel fibrillar array never formed in the CCL with degraded primary bundles, whereas activated fibrocytes constantly underwent chondroid metaplasia. The pathogenic mechanism underlying chondroid metaplasia was thought to be nonspecific and attributable to an essential property of activated fibrocytes in the mature tendon tissue.     

Biodegradation of domestic wastewater under the simulated conditions of Thailand

Serious concerns have been raised about the sustainability of conventional mechanized linear wastewater treatment systems in the developing world. Bangkok in Thailand is no exception to these concerns, and merits closer examination. This paper is aimed at unveiling the fundamental facts and characteristics relating to the organic degradation potentials of domestic wastewater in Bangkok. It is supported by evidence from simple laboratory experiments in which existing local conditions were simulated. The results showed that greywater‐like qualities of domestic wastewater do predominate in Bangkok, most probably as a result of three main factors: (1) continued use of leaching septic tanks, (2) the tradition of using water after defecation, instead of toilet paper, and (3) high organic degradation potentials under tropical monsoon conditions. Considering the small organic content, coupled with high organic degradation rates of the freshly collected greywater‐based sewage in Bangkok, low‐cost and decentralized natural treatment systems could be far more suitable for environmental sustainability in such tropical monsoon areas than the implementation of large‐scale sewage works.     

Identification and partial characterization of a glycoprotein species with high affinity for methylmercury in peripheral nervous tissues of man and experimental animals

A small amount of a glycoprotein species (21-kDa glycoprotein) with high affinity for methylmercury (MeHg) was detected in the post-nuclear or post-mitochondrial supernatant fraction of the homogenate of rat sciatic nerve on electrophoresis and autoradiography after binding of Me203Hg to the fraction. The 21-kDa glycoprotein was also found in the subcellular fractions of mouse, hamster, guinea pig, rabbit and human peripheral nervous tissues. Experiments with the cellular fractions of the tissues revealed that the 21-kDa glycoprotein is localized mainly in the myelin fraction, whereas it was not found in the cellular fractions of brain, spinal cord and nonneural tissues, such as kidney and liver. The specific binding activity of the 21-kDa glycoprotein with MeHg was 12-15 fold that of the major myelin protein, Po. It was shown that the interaction of the 21-kDa glycoprotein with MeHg was mediated through sulfhydryl groups in experiments with iodoacetamide and dithiothreitol. The amino acid compositions of the rat and human 21-kDa glycoproteins were similar but very different from that of a typical metallothionein. The N-terminal amino acid sequences of the two components of the rat 21-kDa glycoprotein were identical to those of P0 and PMP-22, respectively. The in vitro binding of MeHg was also observed in the myelin fraction obtained from the sciatic nerves of MeHg-dosed rats.     

The vector potential revisited

The vector potential in electrodynamics is investigated through the decomposition of its form to the following two parts: 1) the so-called transverse part represented by a divergenceless vector; and 2) the longitudinal part represented by an irrotational vector. The decomposition can be done by the Helmholtz theorem in the vector analysis because the conditions which should be required when the Helmholtz theorem is used are satisfied for the almost vector potentials of physically interesting problems. As an example of such interesting problems, the Aharonov-Bohm effect is chosen here. As for the Aharonov-Bohm effect, the vector potential given in the original paper of Aharonov and Bohm has the singularities along the z-axis. It is shown that even for such a singular potential the Helmholtz theorem is held provided that the concept of the distribution is introduced in it. Generally, the transverse part of the vector potential obtained through such a decomposition is determined uniquely by the magnetic field and does not alter by a gauge transformation. On the other hand, the longitudinal part depends on the choice of special gauge. It is shown that the Aharonov-Bohm effect is due to the contribution of the transverse part of the vector potential and therefore should not be influenced by any gauge transformations.     

Kidney-specific expression of a novel mouse organic cation transporter-like protein

OBJECTIVE: To assess the anterior mediastinal mass in recurrent testicular cancer, with relation to thymic hyperplasia after treatment. METHODS: The anterior mediastinal regions were fully evaluated by chest computed tomography (CT) at the initial staging and after treatment in 24 of 44 patients with testicular cancer. RESULTS: One patient with stage IIB tumor had thymic hyperplasia before treatment, and one with stage III had benign thymic hyperplasia after chemotherapy with salvage surgery. Three of 4 patients who had recurrence had an anterior mediastinal mass. One had benign thymic hyperplasia confirmed by histology and 2 had metastatic tumor confirmed by histology and clinical course, in which the mass became so enlarged that it obstructed major vessels. CONCLUSION: Although the relationship of the CT finding to the response to treatment in the anterior mediastinal mass and other metastatic lesions provide some clues helpful in differentiating benign from malignant masses, surgical exploration is recommended for the patient with an indication for salvage surgery.     

Distribution of several gut neuropeptides and their effects on motor activity in muscularis mucosae of guinea-pig proximal colon

The distribution of peptide-containing nerve fibers and the effect of their neuropeptides on motor activity were studied in the muscularis mucosae of the guinea-pig proximal colon. In the immunohistochemical study, it was shown that the tachykinin (TK)-containing nerve fibers densely innervated the muscularis mucosae. In the superfusion study, three kinds of TKs, i.e., neurokinin A (NK-A), neurokinin B (NK-B) or substance P (SP), enhanced the spontaneous activity on the strips of muscularis mucosae with a tetrodotoxin (TTX)-insensitive manner. Their potency was in the rank order of NK-A > SP. This suggests that the muscle has a predominant NK2 receptor. Calcitonin gene-related peptide (CGRP)-immunoreactive fibers were commonly observed in the muscle. CGRP induced a potent inhibition on spontaneous activity and a concentration-dependent inhibition on the NK-A-elicited excitation in the presence of TTX, indicating its direct effect on the receptor in the muscle. On the other had, gastrin releasing peptide (GRP), galanin, neuropeptide Y or somatostatin were more or less immunopositive in nerve fibers, but they had no effect on the motility of the muscle except that GRP sometimes showed a faint increase in spontaneous activity. Neither methionine-enkephalin nor gastrin-17/cholecystokinin was immunoreactive and had any effect on the muscle. These neuropeptides other than TKs and CGRP do not seem to be neuromediators of motor activity of muscularis mucosae. The results suggest the possibility that TK-, especially NK-A- and CGRP-containing neurons, participate in the regulation of motor activity of the muscularis mucosae in the guinea-pig proximal colon.     

Cryoprotective effects of cycloinulohexaose on freezing and freeze-drying of liposomes

Cycloinulohexaose (CF-6) maintained the membrane structure of the liposomes during freeze-thawing, and thus CF-6 was very effective in inhibiting both drug leakage from liposomes and the size change of liposomes during freezing and freeze-drying. As the ratio of CF-6 to lipid increased from 1 to 5, calcein retention in the liposomes increased from 70% up to 85% and a maximum retention was obtained at a ratio of more than 2. The addition of glycerol enhanced the above cryoprotective effect of CF-6 and improved calcein retention up to 90%. Neither detectable leakage of calcein nor particle size change was observed after the storage of liposomes freeze-dried with CF-6 for 6 months at 4 and 25 degrees C.     

Finite element analysis of fracture statistics of ceramics: Effects of grain size and pore size distributions

A novel numerical simulation method based on finite element analysis (FEA), which can evaluate the fracture probability caused by the characteristics of flaw distribution, is considered an effective tool to facilitate and increase the use of ceramics in components and members. In this study, we propose an FEA methodology to predict the scatter of ceramic strength. Specifically, the data on the microstructure distribution (i.e., relative density, size and aspect ratio of pore, and grain size) are taken as the input values and reflected onto the parameters of a continuum damage model via a fracture mechanical model based on the circumferential circular crack emanating from an oval spherical pore. In addition, we numerically create a Weibull distribution based on multiple FEA results of a three‐point bending test. Its validity is confirmed by a quantitative comparison with the actual test results. The results suggest that the proposed FEA methodology can be applied to the analysis of the fracture probability of ceramics.