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Biomedical subjects

Z Y Chen

Publications and source records attributed to Z Y Chen.

At least 19 recordsLinked to original sources
PubMed1997

Essential role of POU-domain factor Brn-3c in auditory and vestibular hair cell development.

The Brn-3 subfamily of POU-domain transcription factor genes consists of three highly homologous members-Brn-3a, Brn-3b, and Brn-3c-that are expressed in sensory neurons and in a small number of brainstem nuclei. This paper describes the role of Brn-3c in auditory and vestibular system development. In the inner ear, the Brn-3c protein is found only in auditory and vestibular hair cells, and the Brn-3a and Brn-3b proteins are found only in subsets of spiral and vestibular ganglion neurons. Mice carrying a targeted deletion of the Brn-3c gene are deaf and have impaired balance. These defects reflect a complete loss of auditory and vestibular hair cells during the late embryonic and early postnatal period and a secondary loss of spiral and vestibular ganglion neurons. Together with earlier work demonstrating a loss of trigeminal ganglion neurons and retinal ganglion cells in mice carrying targeted disruptions in the Brn-3a and Brn-3b genes, respectively, the Brn-3c phenotype reported here demonstrates that each of the Brn-3 genes plays distinctive roles in the somatosensory, visual, and auditory/vestibular systems.

Animals↗
PubMed1997

The genomic structure of the gene defective in Usher syndrome type Ib (MYO7A).

Usher syndrome type Ib is a recessive autosomal disorder manifested by congenital deafness, vestibular dysfunction, and progressive retinal degeneration. Mutations in the human myosin VIIa gene (MYO7A) have been reported to cause Usher type Ib. Here we report the genomic organization of MYO7A. An STS content map was determined to discover the YAC clones that would cover the critical region for Usher syndrome type Ib. Three of the YACs (802A5, 966D6, and 965F10) were subcloned into cosmids and used to assemble a preliminary cosmid contig of the critical region. Part of the gene encoding human myosin VIIa was found in the preliminary cosmid contig. A cosmid, P1, PAC, and long PCR contig that contained the entire MYO7A gene was assembled. Primers were designed from the composite cDNA sequence and used to detect intron-exon junctions by directly sequencing cosmid, P1, PAC, and genomic PCR DNA. Alternatively spliced products were transcribed from the MYO7A gene: the largest transcript (7.4 kb) contains 49 exons. The MYO7A gene is relatively large, spanning approximately 120 kb of genomic DNA on chromosome 11q13.

Abnormalities, Multiple↗
PubMed1997

Inhibitory effects of jasmine green tea epicatechin isomers on free radical-induced lysis of red blood cells.

Jasmine green tea is an excellent source of natural polyphenol antioxidants including mainly (-) epicatechin (EC), (-) epicatechin gallate (ECG), (-) epigallocatechin (EGC) and (-) epigallocatechin gallate (EGCG). The present study was to test our hypothesis that ingestion of jasmine tea would protect red blood cell (RBC) membrane from free radical-induced oxidation if jasmine tea epicatechin isomers could be absorbed and circulated in blood. When incubated with RBC suspension, all four epicatechin isomers purified from jasmine tea exhibited a strong protection for RBC membrane to hemolysis induced by 2,2'-azo-bis(2-amidinopropane) dihydrochloride (AAPH), an azo free radical initiator. The inhibitory effect was dose-dependent at the concentrations of 2.5 microM to 40 microM. The fatty acid analysis revealed that all four epicatechin isomers significantly prevented loss of arachidonic acid (20:4n-6) and docosahexaenoic acid (22:6n-3) in RBC incubated under the same conditions. Although the in vitro antioxidative activity of EGCG and ECG was more effective than EGC and EC, the latter two isomers were more important in vivo in scavenging free radicals. This was because only EGC and EC instead of EGCG and ECG were circulating in blood stream after a gavage-dose of 100 mg jasmine tea GTP mixture. In fact, ingestion of jasmine tea GTP extracts was associated with a significant decrease in susceptibility of RBC to hemolysis in rats.

Amidines↗
PubMed1997

Cloning and overexpression of two cDNAs encoding the low-CO2-inducible chloroplast envelope protein LIP-36 from Chlamydomonas reinhardtii.

Chlamydomonas reinhardtii, a unicellular green alga, grows photoautotrophically at very low concentrations of inorganic carbon due to the presence of an inducible CO2-concentrating mechanism. During the induction of the CO2-concentrating mechanism at low-CO2 growth conditions, at least five polypeptides that are either absent or present in low amounts in cells grown on high-CO2 concentrations are induced. One of these induced polypeptides with a molecular mass of 36 kD, LIP-36, has been localized to the chloroplast envelope. The protein was purified and the partial internal amino acid sequences were obtained through lys-C digestion. Two cDNAs encoding LIP-36 have been cloned using degenerate primers based on the amino acid sequences. The two genes encoding LIP-36 are highly homologous in the coding region but are completely different in the 5'-end and 3'-end untranslated regions. The deduced protein sequences show strong homology to the mitochondrial carrier protein superfamily, suggesting that LIP-36 is a chloroplast carrier protein. The regulation of the expression of these two genes at high- and low-CO2 growth conditions is also different. Both genes were highly expressed under low-CO2 growth conditions, with the steady-state level of LIP-36 G1 mRNA more abundant. However, neither gene was expressed at high-CO2 growth conditions. The gene products of both clones expressed in Escherichia coli were recognized by an antibody raised against LIP-36, confirming that the two cDNAs indeed encode the C. reinhardtii chloroplast envelope carrier protein LIP-36.

Amino Acid Sequence↗
PubMed1997

Depletion of linoleate induced by weight cycling is independent of extent of calorie restriction.

Recent epidemiological studies have suggested that weight cycling induced by repeated dieting over time may increase the risk of cardiovascular disease. It is speculated that the increased mortality from coronary heart disease for people with a history of excessive weight cycling could be attributed to change in lipid metabolism. Previous studies have demonstrated that repeated cycling of 100% food restriction followed by ad libitum refeeding caused a depletion of linoleate and alpha-linolenate in rats. The objective of the present study was to test the hypothesis that the weight cycling-induced reduction in linoleate and alpha-linolenate is independent of extent of calorie restriction. Two consecutive weight cycles in three experiments were induced by 100% calorie restriction, 60% calorie restriction, and 36% calorie restriction, respectively, followed by ad libitum refeeding. As the consequence of the two weight cycles, linoleate and linolenate were decreased, whereas myristate, palmitate, and palmitoleate were proportionally increased in carcass and adipose tissue lipids. The results of all three experiments showed a preferential depletion of linoleate and alpha-linolenate without changes in final body weight, total body fat, and adipose tissue pads in the weight-cycled rats. In addition, the triacylglycerol species profile in the adipose tissue of weight-cycled rats was significantly remodeled, with a proportional depletion of linoleate-enriched triacylglycerol species (LLL, LLO, and LLP, where L, O, and P are linoleic, oleic, and palmitic acid, respectively) and a proportional accumulation of palmitate-enriched triacylglycerol species (OPPo, PPPo, and PPP, where Po is palmitoleic acid). We conclude that weight cycling changes the ratio of polyunsaturated fatty acids to saturated fatty acids and remodels the adipose tissue triacylglycerol species profile in rats.

Adipose Tissue↗
PubMed1996

Mapping of unconventional myosins in mouse and human.

Myosins are molecular motors that move along filamentous actin. Seven classes of myosin are expressed in vertebrates: conventional myosin, or myosin-II, as well as the 6 unconventional myosin classes-I, -V, -VI, -VII, -IX, and -X. We have mapped in mouse 22 probes encompassing all known unconventional myosins and, as a result, have identified 16 potential unconventional myosin genes. These genes include 7 myosins-I, 2 myosins-V, 1 myosin-VI, 3 myosins-VII, 2 myosins-IX, and 1 myosin-X. The map location of 5 of these genes was identified in human chromosomes by fluorescence in situ hybridization.

Animals↗
PubMed1996

Molecular cloning and domain structure of human myosin-VIIa, the gene product defective in Usher syndrome 1B.

Myosin-VIIa is an unconventional myosin with relatively restricted expression. Cloned first from an intestinal epithelium cell line, it occurs most notably in the testis, in the receptor cells of the inner ear, and in the pigment epithelium of the retina. Defects in myosin-VIIa cause the shaker-1 phenotype in mice and Usher syndrome 1B in human, which are characterized by deafness, lack of vestibular function, and (in human) progressive retinal degeneration. Because the described cDNAs encode less than half of the protein predicted from immunoblots, we have cloned cDNAs encoding the rest of human myosin-VIIa. Two transcripts were found, one encoding the predicted 250-kDa protein and another encoding a shorter form. Both transcripts were found in highest abundance in testis, although the shorter transcript was much less abundant. Both could be detected in lymphocytes by RT-PCR. The myosin tail encoded by the long transcript includes a long repeat of approximately 460 amino acids. Each repeat contains a novel "MyTH4" domain similar to domains in three other myosins, and a domain similar to the membrane-associated portion of talin and other members of the band-4.1 family.

Alternative Splicing↗
PubMed1996

New approach for inhibiting Rev function and HIV-1 production using the influenza virus NS1 protein.

The Rev protein of HIV-1, which facilitates the nuclear export of HIV-1 pre-mRNAs, has been a target for antiviral therapy. Here we describe a new strategy for inhibiting Rev function and HIV-1 replication. In contrast to previous approaches, we use a wild-type rather than a mutant Rev protein and covalently link this Rev sequence to the NS1 protein of influenza A virus, a protein that inhibits the nuclear export of mRNAs. The NS1 protein contains an RNA-binding domain mutation (RM), so that the only functional RNA-binding domain in the chimeric protein (NS1RM-Rev) is in the Rev protein sequence. In the presence of the NS1RM-Rev chimeric protein, HIV-1 pre-mRNAs were retained in, rather than exported from, the nucleus. In addition, this chimeric protein effectively inhibited Rev function in trans in transfection experiments and effectively inhibited the production of HIV-1 in tissue culture cells transfected with an infectious molecular clone of HIV-1 DNA. The inhibitory activities of the NS1RM-Rev chimera were at least equivalent to those of the Rev M10 mutant protein, which has been considered to be the prototype trans inhibitor of Rev function and is currently in phase I clinical trials for the treatment of AIDS patients. We discuss (i) the potential for increasing the inhibitory activity of NS1-Rev chimeras against HIV-1 and (ii) the need for additional studies to evaluate these chimeras for the treatment of AIDS.

Antiviral Agents↗
PubMed1996

Antioxidant activity of natural flavonoids is governed by number and location of their aromatic hydroxyl groups.

We studied antioxidant activity of five natural flavonoids in canola oil, an emulsion, and rat red blood cell membrane. In the canola oil heated at 105 degrees C, myricetin most effectively attenuated lipid oxidation, followed by quercetin, morin and kaempferol in a decreasing order. When the emulsion containing canola oil was incubated at 54 degrees C, quercetin was most protective against lipid oxidation, followed by kaempferol, myricetin and morin in a decreasing rank. Apigenin demonstrated no antioxidant activity in either the canola oil or the emulsion. Of the flavonoids tested, quercetin was the best in quenching free radical chain reaction in rat red blood cell membrane. The antioxidative characteristics of these flavonoids was determined by multiple factors including the system used, the hydrophobicity, and the total number and the location of aromatic hydroxyl groups. We conclude that myricetin is an effective antioxidant in canola oil, while quercetin is an effective protector against lipid oxidation in the cell membrane.

Animals↗
PubMed1996

Isolation of cDNA clones of genes induced upon transfer of Chlamydomonas reinhardtii cells to low CO2.

Unicellular algae grow well under limiting CO2 conditions, aided by a carbon concentrating mechanism (CCM). In C. reinhardtii, this mechanism is inducible and is present only in cells grown under low CO2 conditions. We constructed a cDNA library from cells adapting to low CO2, and screened the library for cDNAs specific to low CO2-adapting cells. Six classes of low CO2-inducible clones were identified. One class of clone, reported here, represents a novel gene associated with adaptation of cells to air. A second class of clones corresponds to the air-inducible periplasmic carbonic anhydrase I (CAH1). These clones represent genes that respond to the level of CO2 in the environment.

Animals↗
PubMed1996

Trans, n-3, and n-6 fatty acids in Canadian human milk.

The presence of trans fatty acids in human milk may be a concern because of their possible adverse nutritional and physiological effects on the recipient infant. The mother's diet is the source of human milk trans fatty acids, and since these fatty acids are prevalent in many common foods of the Canadian diet, the trans fatty acid content and the fatty acid composition of Canadian human milk were measured by gas-liquid chromatography coupled with silver nitrate-thin layer chromatography. In samples obtained from 198 lactating mothers across Canada, the average percentage of total trans (sum of t18:1, t18:2, and t18:3) was 7.2% of breast milk fatty acids with a range of 0.1-17.2%. Analysis of t18:1 isomer distribution indicated that partially hydrogenated vegetable oils are the major source of these trans fatty acids in human milk, whereas contribution from dairy products appeared to be relatively minor. Linoleic and alpha-linolenic acid levels were inversely related to the total trans fatty acids, indicating that the elevation of trans fatty acids in Canadian human milk is at the expense of n-3 and n-6 essential fatty acids. Levels of arachidonic and docosahexaenoic acids did not correlate with their parent fatty acids, indicating that it might be difficult to elevate the levels of n-6 and n-3 c20-22 polyunsaturated fatty acids in breast milk by increasing levels of linoleic and alpha-linolenic acids in the mother's diet.

Canada↗
PubMed1996

Characterization and mapping of the mouse NDP (Norrie disease) locus (Ndp).

Norrie disease is a severe X-linked recessive neurological disorder characterized by congenital blindness with progressive loss of hearing. Over half of Norrie patients also manifest different degrees of mental retardation. The gene for Norrie disease (NDP) has recently been cloned and characterized. With the human NDP cDNA, mouse genomic phage libraries were screened for the homolog of the gene. Comparison between mouse and human genomic DNA blots hybridized with the NDP cDNA, as well as analysis of phage clones, shows that the mouse NDP gene is 29 kb in size (28 kb for the human gene). The organization in the two species is very similar. Both have three exons with similar-sized introns and identical exon-intron boundaries between exon 2 and 3. The mouse open reading frame is 393 bp and, like the human coding sequence, is encoded in exons 2 and 3. The absence of six nucleotides in the second mouse exon results in the encoded protein being two amino acids smaller than its human counterpart. The overall homology between the human and mouse NDP protein is 95% and is particularly high (99%) in exon 3, consistent with the apparent functional importance of this region. Analysis of transcription initiation sites suggests the presence of multiple start sites associated with expression of the mouse NDP gene. Pedigree analysis of an interspecific mouse backcross localizes the mouse NDP gene close to Maoa in the conserved segment, which runs from CYBB to PFC in both human and mouse.

Abnormalities, Multiple↗
PubMed1996

Accumulation of polyunsaturates is decreased by weight-cycling: whole-body analysis in young, growing rats.

Whole-body fatty acid analysis in rats has previously shown that 50-70% of dietary linoleate and alpha-linolenate is beta-oxidized to CO2 and that this value increases with refeeding after a single episode of fasting. Our hypothesis was that repeated fasting-refeeding or weight-cycling would increase the beta-oxidation of linoleate and alpha-linolenate thereby depleting their whole-body levels. In rats consuming 3% energy as linoleate and 0.15% energy as alpha-linolenate during a 16 d balance period, 19% of the linoleate consumed accumulated in weight-cycled rats compared with 34% in the free-fed controls (P < 0.01). Similarly, 11% of the alpha-linolenate consumed accumulated in the weight-cycled rats compared with 22% in the controls (P < 0.01). Arachidonate and docosahexaenoate also accumulated to lower extents in the weight-cycled rats than in the controls. In contrast, whole-body accumulation of palmitate, stearate and oleate was not different between the weight-cycled group and the controls when measured as a proportion of intake or relative to weight gain. Thus, whole-body depletion of linoleate and alpha-linolenate did not occur per se but the partitioning of linoleate and alpha-linolenate was significantly altered by weight-cycling resulting in lower whole-body accumulation and higher apparent oxidation of all polyunsaturates especially linoleate and alpha-linolenate.

Animals↗
PubMed1996

A low-CO2-inducible gene encoding an alanine: alpha-ketoglutarate aminotransferase in Chlamydomonas reinhardtii.

At low-CO2 (air) conditions, the unicellular green alga Chlamydomonas reinhardtii acquires the ability to raise its internal inorganic carbon concentration. To study this adaptation to low CO2, cDNA clones induced under low-CO2 growth conditions were selected through differential screening. One full-length clone is 2552 bp, with an open reading frame encoding 521 amino acids. The deduced amino acid sequence shows about 50% identity with alanine: alpha-ketogutarate aminotransferase (Ala AT, EC 2.6.1.2) from plants and animals, and the mRNA of this clone increased 4- to 5-fold 4 h after cells were switched from high-CO2 to low-CO2 growth conditions. The expression of the enzyme and its activity also increased accordingly at low-CO2 growth conditions. To study the physiological role of Ala AT, a pyridoxal phosphate inhibitor, aminooxyacetic acid, was added at 40 microM to the growth medium when cells were beginning to adapt to low CO2. This caused a 30% decrease in the maximum photosynthetic rate in air-adapting cells 8 h later. The addition of the inhibitor also caused the cells to excrete glycolate, a photorespiratory intermediate, but did not change the apparent affinity of the cell for external CO2. These physiological studies are consistent with the assumption that Ala AT is involved in the adaptation to low-CO2 conditions.

Adaptation, Biological↗
PubMed1996

Myosin VIIA mutation screening in 189 Usher syndrome type 1 patients.

Usher syndrome type 1b (USH1B) is an autosomal recessive disorder characterized by congenital profound hearing loss, vestibular abnormalities, and retinitis pigmentosa. The disorder has recently been shown to be caused by mutations in the myosin VIIa gene (MYO7A) located on 11q14. In the current study, a panel of 189 genetically independent Usher I cases were screened for the presence of mutations in the N-terminal coding portion of the motor domain of MYO7A by heteroduplex analysis of 14 exons. Twenty-three mutations were found segregating with the disease in 20 families. Of the 23 mutations, 13 were unique, and 2 of the 13 unique mutations (Arg212His and Arg212Cys) accounted for the greatest percentage of observed mutant alleles (8/23, 31%). Six of the 13 mutations caused premature stop codons, 6 caused changes in the amino acid sequence of the myosin VIIa protein, and 1 resulted in a splicing defect. Three patients were homozygotes or compound heterozygotes for mutant alleles; these three cases were Tyr333Stop/Tyr333Stop, Arg212His-Arg302His/Arg212His-Arg302His, and IVS13nt-8c-->g/Glu450Gln. All the other USH1B mutations observed were simple heterozygotes, and it is presumed that the mutation on the other allele is present in the unscreened regions of the gene. None of the mutations reported here were observed in 96 unrelated control samples, although several polymorphisms were detected. These results add three patients to single case reported previously where mutations have been found in both alleles and raises the total number of unique mutations in MYO7A to 16.

Alleles↗
PubMed1995

Fasting/refeeding has little lasting effect on long-chain fatty acids accumulating in the developing rat conceptus.

The objective of this study was to delineate the influence of maternal fasting-refeeding on accumulation of long-chain fatty acids by the developing conceptus in chow-fed pregnant rats. At term, the rat conceptus normally accumulates < 5% of total maternal whole body n - 6 polyunsaturates and < 15% of whole body n - 3 polyunsaturates. In contrast to the ad libitum-fed controls, the conceptus of fasted-refed rats was the main site of accumulation of n - 6 and n - 3 polyunsaturates since the maternal carcass (excluding liver) actually had a lower content of polyunsaturates at the end of the fasting-refeeding period than at the start. The rise in the content of triacylglycerols in the conceptus during fasting was specifically attributable to increased arachidonate, docosatetraenoate and docosahexaenoate; all changes in conceptus triacylglycerols were back to ad libitum control values by the end of the refeeding period (term). Conversely, although conceptus total phospholipids did not change significantly during fasting-refeeding, there was a net increase in the phospholipid content of both palmitate and oleate in conceptus after refeeding. We conclude that the developing rat conceptus is largely resistant to the marked changes in maternal fatty acid intake and retention that occur during fasting and refeeding.

Animals↗
PubMed1995

A bidirectional YAC walk from the Norrie disease (NDP) locus.

The region of Xp between DXS7 and the centromere contains the gene for Norrie disease in addition to the genes for several other ophthalmic disorders. A 650-kb YAC containing the loci MAOA, MAOB, and NDP has been used as the starting point for a bidirectional chromosomal walk. A contig of 16 YACs covering between 2 and 3 Mb has been developed in which the following markers/genes are located (in physical order): Xpter--DXS1201 (256ze5)--DXS6668--DXS228--DXS77--MAOA--++ +MAOB--FR12 (pseudogene)--NDP--DXS6670--RRM2P3--DXS6671--DXS742 --Xcen. Seven new STSs are described both for end clones and for internal Alu PCR products from the contig. The contig contains the breakpoint of the t75-2ma-1b (t75) translocation, close to the 5' end of the MAOB gene.

Base Sequence↗