Effects of Particle Size of Bulk Drug and food on the Bioavailability of U-78875 in Dogs

The effects of particle size and food on the absolute bioavailability of U-78875 in dogs after oral administration of either a suspension or tablet dosage form were investigated. A reduction of particle size caused a significant increase in bioavailability along with an increase in dissolution rate. Additionally, both suspension and tablet dosage forms administered after food caused an increase in bioavailability. Thus, to accelerate drug dissolution, a reduction of U-78875 particle size from the unmilled state is important for the optimization of formulation compositions. To increase the bioavailability of U-78875, postprandial dosing should be considered.     

Mutational analysis of structure--activity relationships in human tumor necrosis factor-alpha

To determine the region of human tumor necrosis factor-alpha(TNF-), essential for cytotoxic activity against mouse L-M cells,single amino-acid-substituted TNF- mutant proteins (muteins)were produced in Escherichia coli by protein engineering techniques.An expression plasmid for TNF- was mutagenized by passage throughan E.coli mutD5 mutator strain and by oligonucleotide-directedmutagenesis. Approximately 100 single amino-acid-substitutedTNF- muteins were produced and assayed for cytotoxic activity.The cytotoxic activities of purified TNF- muteins, e.g. TNF-31T,-32Y, -82D, -85H, -115L, -141Y, -144K and -146E, were < 1%of that of parent TNF-. These results indicate that the integrityof at least four distinct regions of the TNF- molecule is requiredfor full biological activity. These regions are designated asfollows: region I, from position 30 to 32; region II, from position82 to 89; region III, from position 115 to 117; region FV, fromposition 141 to 146. In addition, TNF-141Y could not completelycompete with parent TNF- for binding to the receptor. This demonstratesthat region IV, and at least aspartk acid at position 141, mustbe involved in the TNF receptor binding site.     

Defective bone remodelling in osteoprotegerin-deficient mice

Previous studies have reported enhanced osteoclastogenesis, increased bone resorption and osteoporosis in osteoprotegerin (OPG)-deficient mice. In the present study, we show that the tibial epiphyses contain abundant, thin trabeculae lined with numerous osteoclasts and cuboidal osteoblasts. The increase in osteoblasts and osteoclasts was associated with a dramatic increase in calcein labelling of the mineralization fronts and replacement of much of the intertrabecular marrow with numerous alkaline phosphatase-positive preosteoblasts. Furthermore, the discrete, linear cement lines seen in wild-type mice were replaced by a randomly oriented meshwork of cement lines that were stained intensely for tartrate-resistant acid phosphatase and osteopontin in the OPG-/- mice. These indices of accelerated bone remodelling in mutant bone were associated with irregular trabecular surfaces, a disorganized collagen matrix interspersed with amorphous ground substance and numerous fissures between old and new bone. In total, these observations indicate that enhanced osteoclastic activity in OPG-/- epiphyses led to a coupled increase in osteoblast differentiation and activity and an increase in bone remodelling. The high bone turnover, disorganized matrix and impaired attachment of new to old bone in the cement lines in OPG-/- mice appear to cause bone fragility.     

ZnSe/ZnMgSSe blue laser diode

The continuous wave operation of an ZnSe/ZnMgSSe laser diode was achieved for the first time at 77 K. Blue stimulated emission was observed at a wavelength of 447 nm and the threshold current density was 225 A/cm/sup 2/.<>     

Contribution of high p34cdc2 kinase activity to premature chromosome condensation of injected somatic cell nuclei in rat oocytes

The present study was undertaken to clarify the relationship between the p34cdc2 kinase activity of in vitro-aged or enucleated rat oocytes and the premature chromosome condensation (PCC) of microinjected cumulus cell nuclei. Wistar rat oocytes were placed in vitro up to 120 min after the animal was killed. The p34cdc2 kinase activity of the oocytes decreased in a time-dependent manner. The incidence of PCC was higher when nuclear injection into intact oocytes was completed in 15-45 min rather than 46-120 min. When rat oocytes were enucleated for subsequent nuclear injection, the p34cdc2 kinase activity transiently increased soon after enucleation but drastically decreased after 30 min. Removal of the cytoplasm instead of the meta-phase-plate did not affect the p34cdc2 kinase activity even after 60 min. PCC occurred in intact and cytoplasm-removed oocytes but not in enucleated oocytes. In contrast, oocytes from BDF1 mice exhibited a p34cdc2 kinase level twice that of rat oocytes and supported PCC despite enucleation. The p34cdc2 kinase level of intact rat oocytes was reduced to the equivalent level of aged (120 min) or enucleated (+60 min) oocytes by a 45 min treatment with roscovitine, an inhibitor of p34cdc2 kinase. None of the roscovitine-treated oocytes supported PCC while half of the control oocytes did. When rat oocytes were treated with MG132, a proteasome inhibitor, delayed inactivation of the p34cdc2 kinase was observed in the MG132-treated oocytes. A significantly higher proportion of the MG132-treated oocytes supported PCC when compared with the control oocytes. Moreover, a higher proportion of MG132-treated and enucleated oocytes carried two pseudo-pronuclei after cumulus cell injection and developed to the two-cell stage when compared with the enucleated oocytes at the telophase-II stage. These results suggest that the decreased level of p34cdc2 kinase activity in aged or enucleated rat oocytes is responsible for their inability to support PCC of microinjected donor cell nuclei and that inhibition of p34cdc2 kinase inactivation by chemicals such as MG132 is in part effective for rat oocytes to promote PCC and further development.     

Epidemiological studies of cerebro-and cardiovascular diseases in the northeast of Japan Part I. Pathological approach to the study in Akita Prefecture

To define the etiologic factors of Japanese cerebrovascular diseases, a pathological research was conducted on autopsy cases of stroke in Akita Prefecture. In most cases of cerebral hemorrhages, especially in the middle age group (30-59 of age), arteriosclerotic deviations in basal berebral arteries of the circle of Willis and intracerebral small arteries are not found. The authors conclude that cerebral hemorrhage may occur without relationship to disturbances of lipid metabolism, biochemically, and to atherosclerosis, pathologically. Concerning cerebral infarction, especially in the old generation (over 60 years of age), severe arteriosclerotic deviations were recognized both in basal cerebral and intracerebral arteries. These changes were highly influenced by the grade and duration of hypertension, and rarely influenced by hypercholesterolemia. According to the above-mentioned facts, the results obtained from out epidemiological survey were confirmed by the pathological studies.     

Angiotensin I-converting enzyme inhibitory activities of Chinese fermented soypaste and estimation of the inhibitory substances

Angiotensin I-converting enzyme (ACE) inhibitory activities of aqueous extracts of Chinese commercial soypaste were investigated in this work. Six samples from northern China showed potent ACE inhibitory activities with IC₅₀ values less than 40.0 μg/mL. ACE inhibitors in the sample with the strongest activity were purified by ultrafiltration, solid-phase extraction and gradient RP-HPLC. According to spectroscopic methods, a compound (M328.1) was separated as C₁₅H₂₁NO₇. It was supposed to be a conjugate of phenylalanine and glucose generated by Maillard reaction during soypaste production, providing support on the contribution of Maillard reaction products to the ACE inhibitory activity of the sample. Results further indicated that the total ACE inhibition by the sample occurred from the combined function of various bioactive substances, suggesting that Chinese soypaste could be a good source of ACE inhibitors for exploring functional foods or ingredients with antihypertensive effect.     

Safety and structural analysis of polymers produced in manufacturing process of alpha-lipoic acid

Alpha-Lipoic acid has recently been permitted for use in foodstuffs and is contained in tablets and capsules. Although alpha-lipoic acid is synthesized from adipic acid, the safety of polymers produced during the purification and drying processes has been an issue of concern. Hence, we examined the safety profiles of thermally denatured polymer (LAP-A) and ethanol-denatured polymer (LAP-B) produced in the manufacturing process of alpha-lipoic acid. Furthermore, we conducted structural analysis of these polymers by 1H-NMR and FAB-MS spectroscopy. In a consecutive ingestion test, male and female mice ingested diet containing 0.1 and 0.2% LAP-A and -B for 4 weeks. Blood uric acid, potassium and lactate dehydrogenase (LDH) tended to increase without dose-dependency. Relative liver weights were also increased. However, male dogs that were orally administered LAP-B (500 mg/kg) once did not show any abnormalities in blood parameters or general condition. These findings indicate that alpha-lipoic acid polymers are not acutely toxic; however, chronic ingestion of these polymers may affect liver and kidney functions.     

Characterization of Aspergillus oryzae glycoside hydrolase family 43 β-xylosidase expressed in Escherichia coli

This is the first report of glycoside hydrolase family 43 β-xylosidase from Aspergillus oryzae. To characterize this enzyme, the recombinant enzyme was expressed in Escherichia coli. Unlike known β-xylosidases from fungal origins, the enzyme did not show substrate ambiguity and was stable at alkaline pH.