Identification of three novel missense PKU mutations among Chinese.

Three novel missense mutations have been identified in the phenylalanine hydroxylase (PAH) genes of Chinese individuals afflicted with various degrees of phenylketonuria (PKU). A T-to-C transition was observed in exon 5 of the gene, resulting in the substitution of Phe161 by Ser161. Two substitutions, G-to-T and T-to-G, were observed in exon 7, resulting in the substitution of Gly247 by Val247 and Leu255 by Val255, respectively. Expression analysis demonstrated that these mutant proteins produced between 0 and 15% of normal PAH enzyme activity. Population screening of a Chinese sample population indicates that these mutations are quite rare, together accounting for only about 4% of all PKU alleles among the Chinese. The P161S and G247V mutations were each present on a single PAH RFLP haplotype 4 chromosome in patients form Northern China, while the L255V mutation was present on chromosomes of both haplotypes 18 and 21 in patients from Southern China. These results suggest that the remaining 30% of uncharacterized PKU alleles in the Chinese population may bear a large number of relatively rare PAH mutations.     

In vitro effects of fetal bovine serum and glucose on multiplication of Cryptobia salmositica

Cryptobia salmositica multiplied rapidly at 10 C in a minimum essential medium (containing 1.0 mg glucose/ml, Hanks' salts and L-glutamine) supplemented with heat-inactivated fetal bovine serum (FBS) and HEPES buffer (25 mM). The multiplication rate of C. salmositica was related to the amount of FBS; the peak number (approximately 9 x 10(6) parasites/ml) was attained in about 6 wk when the medium contained 25% final concentration of FBS. Glucose utilization was related to the number of parasites; the maximum utilization was reached before peak numbers. Formation of rosette colonies was correlated with multiplication rate and numbers of parasites in cultures. Degenerate round forms found in old cultures probably were caused by the accumulation of metabolic wastes in the medium.     

A scanning electron microscope study on the tegument of Proalarioides kobayashii Park, 1940 (Trematoda)

A SEM study was performed on the surface of adult P. kobayashii Park, 1940, recovered from the snake, Elaphe rufodorsata. The anterior part of the worms was cup-shape and equipped with oral, ventral suckers, pseudosuckers, and tribocytic organ, and the posterior one was finger-like and round-ended. The tegument of the anterior body was covered with 3-4 pointed small spines on the mid-ventral surface and 1-2 pointed ones on the lateral surface. Sensory papillae such as type II, dome-shape ones, and papillae with an opening were distributed over the ventral surface of the anterior portion. The round tribocytic organ was bearing small stout spines laterally, whereas the surface which comes in contact with the host tissues consisted of numerous long fibrillar fibers. The lip of the oral sucker contained type II papillae. Lateral margin of the anterior body revealed type III papillae.     

Lysophosphatidylserine enhances the transfer of 22:6n3 to lysophosphatidic acid in rat brain microsomes.

Although the acyl groups of phosphatidylserine in brain are uniquely enriched in docosahexaenoic acid (22:6n3), the mechanism for this enrichment is not well understood. When rat brain homogenates and microsomes were incubated in the presence of lysophosphatidylserine (LPS) together with [14C]22:6n3 and cofactors for activation to its acylCoA, very little radioactivity was incorporated into phosphatidylserine (PS). On the other hand, [14C]20:4n6 was more actively incorporated into PS. Addition of LPS (1-10 uM), however, resulted in a 2-5 fold enhancement of the transfer of labeled 22:6n3 and 20:4n6 to phosphatidic acid (PA). Kinetic analysis indicated the ability of LPS to lower the Km and increase the Vmax of the lysophosphatidic acid (LPA) acyltransferase reaction. Among other lysophospholipids tested, lysophosphatidylserine was most effective in enhancing PA biosynthesis. Since PA is an important intermediate for de novo biosynthesis of phospholipids, these results reveal a novel mechanism for promoting synthesis of PA enriched in polyunsaturated fatty acids in brain.     

Conformational epitope on gp120 important in CD4 binding and human immunodeficiency virus type 1 neutralization identified by a human monoclonal antibody.

A human monoclonal antibody designated 15e is reactive with the envelope glycoprotein (gp120) of multiple isolates of human immunodeficiency virus type 1 (HIV-1). Antibody 15e also neutralizes HIV-1 with broad specificity and blocks gp120 binding to CD4. Characterization of the 15e epitope shows that it is conformation dependent and is distinct from previously recognized functional domains of gp120, suggesting that this epitope represents a novel site important for HIV-1 neutralization and CD4 binding. These findings have implications for the development of a vaccine for AIDS.     

Isolation and Characterization of a UDPGlucose: Flavonol O-Glucosyltransferase from Illuminated Red Cabbage (Brassica oleracea cv Red Danish) Seedlings

A UDPGlc:flavonol O³-glucosyltransferase (EC 2.4.1.91) that catalyzes the formation of quercetin and kaempferol O³-glucosides has been purified about 1450-fold from illuminated red cabbage (Brassica oleracea cv Red Danish) seedlings with a 3.3% yield. Purification of the enzyme was achieved by (NH₄)₂SO₄-precipitation, gel-filtration, ion-exchange chromatography on DEAE-Bio-Gel and Q-Sepharose, chromatofocusing, and electrophoresis in nondenaturing polyacrylamide (10%) gels. The enzyme preparation had a pH optimum between 5.8 and 6.2, isoelectric point in the pH range 4.25 to 4.55, a M_r of 59,000, and it was composed of two similar subunits of M_r 29,500. The glucosyltransferase reached half substrate saturation at 180 micromolar (UDPGlc) and 7 micromolar (quercetin) concentrations. Kaempferol, which was glucosylated at a relative rate of 87%, had a lesser affinity for the enzyme (K_m~12 micromolar). Flavanones, flavanols, flavones, dihydroflavonols, and anthocyanidins were not readily utilized as substrates, suggesting that the enzyme is specific for flavonol glucoside biosynthesis.     

Abscisic Acid induces anaerobiosis tolerance in corn

Flooding is a frequently occurring environmental stress that can severely affect plant growth. This study shows that treatment of corn (Zea mays L.) seedlings with abscisic acid (ABA) increases their tolerance to anoxia 10-fold over untreated seedlings and twofold over seedlings treated with water. Corn seedlings stressed anoxically for 1 day showed only 8% survival when planted in vermiculite. Pretreatment of root tips with 100 micromolar ABA or water for 24 hours before the 1 day anoxic stress increased the anoxic survivability of seedlings to 87% and 47%, respectively. Cycloheximide (5 milligrams per liter), added together with ABA, reduced the seedling survival rate, indicating that the induction of anoxic tolerance in corn by ABA was partly a result of the synthesis of new proteins. ABA treatment induced a threefold increase in alcohol dehydrogenase enzyme activity in corn roots. However, after 24 h of anoxia, alcohol dehydrogenase enzyme activity between the ABA-pretreated and non-pretreated corn roots was not significantly different. The results indicated that ABA played an important role in inducing anoxic tolerance in corn and that the induced tolerance was probably mediated by an increase in alcohol dehydrogenase enzyme activity before the anoxic stress.     

Field uniformity of the Japonica rice region of Taiwan as estimated by relative genetic contribution

Summary Despite the concerns for genetic vulnerability that were raised in the 1970s, the field uniformity of the Japonica rice (Oryza sativa L.) region in Taiwan has increased since 1980 with over 82% of the cultivated areas being covered by as few as three varieties and over half of this hectarage by a single variety. Japanese plant introductions are the major ancestral contributors of genetic constituents for varieties released in Taiwan. The main constitution of the genetic base present in the field has changed little since 1971. Six common ancestors comprised 60%, 55%, 78%, and 77% of the genetic constituents present in the field in 1971, 1976, 1981, and 1986, respectively. These estimates revealed that at least 55% of the genes utilized in the last 15 years came from the same sources. Recent efforts in introducing new germ plasm sources to variety development should continue to alleviate the possible crop loss due to continuous monoculture.Research supported by National Science Council (NSC 78-0211-B005-14)     

The effects of knee motion and external loading on the length of the anterior cruciate ligament (ACL): a kinematic study

A six-degrees-of-freedom mechanical linkage device was designed and used to study the unconstrained motion of ten intact human cadaver knees. The knees were subjected to externally applied varus and valgus (V-V) moments up to 14 N-m as well as anterior and posterior (A-P) loads up to 100 N. Tests were done at four knee flexion angles; 0, 30, 45, and 90 deg. Significant coupled axial tibial rotation was found, up to 21.0 deg for V-V loading (at 90 deg of flexion) and 14.2 deg for A-P loading (at 45 deg of flexion). Subsequently, the knees were dissected and the locations of the insertion sites to the femur and tibia for the anteromedial (AM), posterolateral (PL), and intermediate (IM) portions of the ACL were identified. The distances between the insertion sites for all external loading conditions were calculated. In the case when the external load was zero, the AM portion of the ACL lengthened with knee flexion, while the PL portion shortened and the intermediate (IM) portion did not change in length. With the application of 14 N-m valgus moment, the PL and IM portions of the ACL lengthened significantly more than the AM portion (p less than 0.001). With the application of 100 N anterior load, the AM portion lengthened slightly less than the PL portion, which lengthened slightly less than the IM portion (p less than 0.005). In general, the amount of lengthening of the three portions of the ACL during valgus and anterior loading was observed to increase with knee flexion angle (p less than 0.001).     

Identification of the posttranslational modifications of bovine lens alpha B-crystallins by mass spectrometry.

A combination of mass spectrometric techniques has been used to investigate the amino acid sequence and post-translational modifications of alpha B-crystallin isolated from bovine lenses by gel filtration chromatography and reversed-phase high performance liquid chromatography. Chromatographic fractions were analyzed by electrospray ionization mass spectrometry to determine the homogeneity and molecular weights of proteins in the fractions. The alpha B-crystallin primary gene product, its mono- and diphosphorylated forms, its N- and C-terminal truncated forms, as well as other lens proteins unrelated to the alpha B-crystallins were identified by their molecular weights. Detailed information about the sites of phosphorylation, as well as evidence supporting reassignment of Asn to Asp at position 80, was obtained by analyzing proteolytic digests of these proteins by fast atom bombardment mass spectrometry. Results of this investigation indicate that alpha B-crystallin is phosphorylated in vivo at Ser 45, Ser 59, and either Ser 19 or 21. From the specificity of phosphorylation of alpha-crystallins, it appears that there may be two different kinases responsible for their phosphorylation.