Association and haplotype analysis of purH gene with inosine monophosphate content in chickens

The current study was designed to investigate the effects of the purH gene on chicken muscle inosine monophosphate (IMP) content. Muscle IMP content was measured in five chicken breeds. Single nucleotide polymorphisms (SNPs) were detected by PCR-SSCP and DNA sequencing. Two SNPs were detected, A/T substitution at position 8023 in exon 9, and T/C substitution at position 17446 in exon 16. The results indicated that only T17446C polymorphism was associated with IMP content. The haplotype effect was higher than the single genotype effect. We tentatively conclude that purH gene is a candidate locus or linked to a major gene that affects muscle IMP content. Haplotypes are superior to single genotypes as potential molecular markers for meat quality traits in chicken.     

Association Between PCR-SSCP of Bone Morphogenetic Protein 15 Gene and Prolificacy in Jining Grey Goats

The Jining Grey is a prolific local goat breed in P.R. China. Bone morphogenetic protein 15 (BMP15) gene that controls high fecundity of Inverdale, Hanna, Lacaune, Belclare, Cambridge, and Small Tailed Han ewes was studied as a candidate gene for the prolificacy of Jining Grey goats. According to the sequence of ovine BMP15 gene, six pairs of primers were designed to detect single nucleotide polymorphisms in exon 1 and exon 2 of the BMP15 gene in both high fecundity breed (Jining Grey goats) and low fecundity breeds (Boer, Liaoning Cashmere, and Inner Mongolia Cashmere goats) by single strand conformation polymorphism (SSCP). Two pairs of primers (F1/R1 and F2/R2) were used to amplify the exon 1. Four pairs of primers (F3/R3, F4/R4, F5/R5, and F6/R6) were used to amplify the exon 2. Only the products amplified by primer F5/R5 displayed polymorphism. Results indicated that two genotypes (AA and AB) were detected in prolific Jining Grey goats and only one genotype (AA) was detected in low fecundity goat breeds. In Jining Grey goats frequencies of genotypes AA and AB were 0.10 and 0.90, respectively. Sequencing revealed two point mutations (G963A and G1050C) of BMP15 gene in the AB genotype in comparison to the AA genotype. In Jining Grey goats the heterozygous AB does had 1.13 (p < 0.01) kids more than the homozygous AA does. These results preliminarily showed that the BMP15 gene is either a major gene that influences the prolificacy of Jining Grey goats or a molecular genetic marker in close linkage with such a gene.     

Association of the Leptin Gene with Carcass Characteristics in Nellore Cattle

Advances in DNA technology have created biotechnological tools that can be used in animal selection and new strategies for increasing herd productivity and quality. The objective of the present work was to associate the genotypes of leptin gene exon 2 polymorphisms with productive traits in Nellore cattle. Blood was collected from Nellore males and PCR-RFLP reactions were performed with the restriction enzymes ClaI and Kpn2I. The gene frequencies resulting from digestion by ClaI were 0.60 and 0.40 for allele A and T, respectively; the genotypic frequencies were AA = 0.20 and AT = 0.80. The gene frequencies from digestion by Kpn2I were 0.81 for allele C and 0.194 for allele T; the genotypic frequencies were CC = 0.62 and CT = 0.38. The populations in both cases were not in Hardy-Weinberg equilibrium (p > 0.05), and the TT genotype was not found. Significant associations were noted between leptin gene exon 2 polymorphisms and five productive traits in Nellore cattle: carcass fat distribution, the intensity of red muscle coloration, pH, marbling, and post-slaughter fat thickness.     

Association Between Sexual Precocity and Alleles of KISS-1 and GPR54 Genes in Goats

KISS-1 and GPR54 were regarded as key regulators for the puberty onset and fundamental gatekeepers of sexual maturation in mammals. To explore the possible association between variations in KISS-1 and GPR54 with sexual precocity, mutation screening of exon 1 of KISS-1 and exon 1, exon 3, and partial exon 5 of GPR54 was performed in a sexual precocious breed (Jining Grey goats) and sexual late-maturing breeds (Inner Mongolia Cashmere, Angora, and Boer goats) by PCR-SSCP. The results showed that five novel mutations were identified in exon 1 and partial exon 5 of GPR54 including C96 T, T173C, G176A, G825A, and C981 T. The Jining Grey goats with genotype BB or AB had 1.07 (P < 0.05) or 0.40 (P < 0.05) kids more than those with AA. The Jining Grey goats with genotype DD or CD had 1.80 (P < 0.05) or 0.55 (P < 0.05) kids more than CC, respectively. The present study preliminarily showed an association between alleles B and D of GPR54 with high litter size and sexual precocity in Jining Grey goats.     

WDNM1 is Associated with Differentiation and Apoptosis of Mammary Epithelial Cells

In this study, we show that expression of the Westmead DMBA8 nonmetastatic cDNA 1 (WDNM1) gene was increased upon SFM and/or TNFα treatment, with a corresponding increase in apoptotic cells, and gradually decreased following re-stimulation with serum in HC11 mammary epithelial cells. TNFα induced WDNM1 expression showed the NFκB-dependent mechanism since it's expression was abrogated in IκBαM (super-repressor of NFκB)-transfected cells, but not those transfected with control vector. Furthermore, overexpression of WDNM1 suppressed growth and differentiation, and accelerated apoptosis of HC11 cells. Thus, our results demonstrate that WDNM1 gene expression, regulated by the TNFα-NFκB signal pathway, is associated with HC11 cell apoptosis.     

A New Tandem Repeat in Bovine Fibrinogen Aα Gene

In this study, we describe intraspecies variation in the αC connector region of the bovine fibrinogen Aα gene. Sequencing and genotyping of six bovine breeds revealed 7 to 10 tandem repeats in the αC connector region. In addition, we observed length differences between B. indicus and B. taurus, with the B. indicus having longer fibrinogen αC connectors (10-repeat alleles) than B. taurus (7- and 9-repeats). The difference in tandem repeats may be related to the function of blood coagulation system.     

A Facile Synthetic Method for 3′-α-Fluoro- 2′,3′-dideoxyadenosine

Abstract

A facile method for the synthesis of 3′-α-fluoro-2′,3′-dideoxyadenosine (5) has been developed using a novel rearrangement of 3′-β-bromine to the 2′-β position during 3′-α fluorination.     

Recent Advances in the Synthesis of Conformationally Locked Nucleosides and Their Success in Probing the Critical Question of Conformational Preferences by Their Biological Targets

Abstract

The present work describes some recent approaches to the syntheses of three classes of locked-North nucleosides: β -D-ribo-, β-D-deoxyribo-, and β-D-dideoxyribonucleosides. The method developed for the latter class permitted access to a novel bicyclo[3.1.0]hexene-type nucleosides structurally similar to D4T and carbovir. A structural analysis and biological activities are discussed.     

A New Synthesis of 9-(2-Deoxy-2-fluoro-β-D-arabinofuranosyl)guanine (araF-G)

Abstract

Interesting and very promising antisense properties of 2′-deoxy-2′-fluoroarabinonucleic acids ((a) Wilds, C.J.; Damha, M.J. 2′-Deoxy-2′-fluoroarabinonucleosides and oligonucleotides (2′F-ANA): synthesis and physicochemical studies. Nucl. Acids Res. 2000, 28, 3625–3635; (b) Viazovkina, E.; Mangos, M.; Elzagheid, M.I.; Damha, M.J. Current Protocols in Nucleic Acid Chemistry 2002, 4.15.1–4.15.21) (2′F-ANA) has encouraged our research group to optimize the synthetic procedures for 2′-deoxy-2′-fluoro-β-D-arabinonucleosides (araF-N). The synthesis of araF-U, araF-T, araF-A and araF-C is straightforward, (Tann, C.H.; Brodfuehrer, P.R.; Brundidge, S.P.; Sapino, C., Jr. Howell H.G. Fluorocarbohydrates in synthesis. An efficient synthesis of 1-(2-deoxy-2-fluoro-β-D-arabinofuranosyl)-5-iodouracil (β-FIAU) and 1-(2-deoxy-2-fluoro-β-D-arabinofuranosyl)thymine (β-FMAU). J. Org. Chem. 1985, 50, 3644–3647; Howell, H.G.; Brodfuehrer, P.R.; Brundidge, S.P.; Benigni, D.A.; Sapino, C., Jr. Antiviral nucleosides. A stereospecific, total synthesis of 2′-fluoro-2′-deoxy-β-D-arabinofuranosyl nucleosides. J. Org. Chem. 1988, 53, 85–88; Maruyama, T.; Takamatsu, S.; Kozai, S.; Satoh, Y.; Izana, K. Synthesis of 9-(2-deoxy-2-fluoro-β-D-arabinofuranosyl)adenine bearing a selectively removable protecting group. Chem. Pharm. Bull. 1999, 47, 966–970) however, the synthesis of the guanine analogue is more complicated and affords poor to moderate yields of araF-G (4) ((a) Elzagheid, M.I.; Viazovkina, E.; Masad, M.J. Synthesis of protected 2′-deoxy-2′-fluoro-β-D-arabinonucleosides. Synthesis of 2′-fluoroarabino nucleoside phosphoramidites and their use in the synthesis of 2′F-ANA. Current Protocols in Nucleic Acid Chemistry 2002, 1.7.1–1.7.19; (b) Tennila, T.; Azhayeva, E.; Vepsalainen, J.; Laatikainen, R.; Azhayev, A.; Mikhailopulo, I. Oligonucleotides containing 9-(2-deoxy-2-fluoro-β-D-arabinofuranosyl)-adenine and -guanine: synthesis, hybridization and antisense properties. Nucleosides, Nucleotides and Nucl. Acids 2000, 19, 1861–1884). Here we describe an efficient synthesis of araF-G (4) that involves coupling of 2-deoxy-2-fluoro-3,5-di-O-benzoyl-α-D- arabinofuranosyl bromide (1) with 2-chlorohypoxanthine (2) to afford 2-chloro-β-araF-I (3) in 52% yield. Nucleoside (3) was transformed into araF-G (4) by treatment with methanolic ammonia (150°C, 6 h) in 67% yield.     

IDENTIFICATION OF CAROTENOIDS PRODUCED FROM CHEESE WHEY BY BLAKESLEA TRISPORA IN SUBMERGED FERMENTATION

The identification of carotenoids in B. trispora during pigment production from deproteinized hydrolyzed whey supplemented with plant oils was studied. The carotenoid content in Blakeslea trispora were β-carotene, γ-carotene, and lycopene. The composition of carotenoids depends of the amount of oils added to the cheese whey. At the maximum concentration of carotenoids, the proportions of β-carotene, γ-carotene, and lycopene (as percent of total carotenoids) was 60.1%, 32.5%, and 7.4%, respectively.     

Comparative Messenger RNA Expression of FSHβ, LHβ, FSHR,LHR, and ERβ in High and Low Prolific Goat Breeds

Follicle-stimulating hormone (FSH) and luteinizing hormone (LH) have a central role in follicle growth and maturation, but no clear differences between breeds with different ovulation rates have been found. Therefore, this study investigated mRNA expression of FSHβ, LHβ, FSH receptor (FSHR), LH receptor (LHR), and estrogen receptor-β (ERβ) genes in prolific Lezhi black (LB) goats and nonprolific Tibetan (TB) goats by real-time PCR. Follicles and pituitaries were recovered from goats at 12–24 h after onset of estrus. Real-time PCR analysis revealed that the expression levels of FSHβ and LHβ mRNA were significantly higher (p < 0.01) in pituitary of LB than in TB does, but the expression levels of FSHR and LHR mRNA in follicle of TB were greater (p < 0.05). Expression level of follicular ERβ was not different between the two breeds. Data provide evidence that the greater ovulation rate in the LB goat as compared to the TB breed is associated with a greater gonadotropin expression during follicular phase.     

A PROCEDURE FOR THE PURIFICATION OF BETA-LACTOGLOBULIN FROM BOVINE MILK USING GEL FILTRATION CHROMATOGRAPHY AT LOW pH

A simple and rapid procedure for the purification of beta-lactoglobulin (β-LG) from bovine milk is described. The procedure exploits the major difference in molecular mass of β-LG and other whey components and the existence of the former in monomeric form at acidic pH. Gel filtration of whey was carried out using a Bio-Gel P10 column at pH 3.0. Residual caseins and other milk proteins were excluded from the gel and β-LG and alpha-lactalbumin (α-LA) emerged as two fully resolved peaks. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) suggested that β-LG was purified to apparent homogeneity, while absorption, fluorescence, and circular dichroism spectroscopy indicated the native-like conformation of the protein. Western blot analysis revealed that the antibodies raised against the purified β-LG in rabbits also readily react with the commercial bovine protein. This procedure requires only 4–5 hr for the purification of about 10 mg of β-LG from a single run while using a small column (2.3 cm × 83 cm) of Bio-Gel P10 and has the potential for scaling up.     

Oligonucleotides Containing Disaccharide Nucleosides: Synthesis,Physicochemical, and Substrate Properties

Abstract

The efficient synthesis of oligonucleotides containing 2′-O-β-D-ribofuranosyl (and β-D-ribopyranosyl)nucleosides, 2′-O-α-D-arabinofuranosyl (and α-L-arabinofuranosyl)nucleosides, 2′-O-β-D-erythrofuranosylnucleosides, and 2′-O-(5′-amino-5-deoxy-β-D-ribofuranosyl)nucleosides have been developed.     

A New Type of Acyclic,Achiral Nucleoside Analogue. How Does It Simulate Nucleosides?

Abstract

The new monomer 1 seems to be an excellent mimic of nucleosides with different sugar conformations (north, south, and envelope), because of the relatively free rotation around γ, δ, and χ. The rotation around χ is primarily controlled by the repulsion between H6 and the two hydrogen atoms on C4′ and not pi conjugation between the double bond and the nucleobase. A viable synthesis of the guanine monomer 8 is described.     

INHIBITION OF SOME HEPATIC GLYCOSIDASES BY THE DISECO NUCLEOSIDE, 4-AMINO-3-(D-GLUCOPENTITOL-1-YL)- 5-MERCAPTO-1,2,4-TRIAZOLE AND ITS 3-METHYL ANALOG

ABSTRACT

The in vivo and in vitro effects of 4-amino-3-(D-glucopentitol-l-yl)-5-mercapto-1,2,4-triazole and its 3-methyl analogue on α- and β-glucosidases, β-glucuronidase as well as α-amylase have been investigated. α-Glucosidase is the enzyme that is markedly affected in vivo and in vitro in a dose-dependent manner. The compounds showed a reversible inhibition of a competitive type for α-glucosidase. Moreover, they exert a relatively potent inhibition on α-glucosidase with a K_i magnitude of 3.6×10^?4, 9.5×10^{? 5} M.     

Synthesis and Biological Evaluation of Some 5-Nitro- and 5-Amino Derivatives of 2′-Deoxycytidine, 2′,3′-Dideoxyuridine,and 2′,3′-Dideoxycytidine

Abstract

In this article, we describe the synthesis of 5-nitro-1-(2-deoxy-α-D-erythro-pentofuranosyl)cytosine (4α), 5-nitro-1-(2-deoxy-β-D-erythro-pentofuranosyl)cytosine (4β), 5-amino-1-(2-deoxy-α-D-erythro-pentofuranosyl)cytosine (5α), 5-nitro-1- (2-deoxy-β-D-erythro-pentofuranosyl)cytosine (5β), 5-nitro-1-(2,3-dideoxy-β- D-ribofuranosyl)uracil (6β), 5-amino-1-(2,3-dideoxy-α,β-D-ribofuranosyl)uracil (7), 5-nitro-1-(2,3-dideoxy-α,β-D-ribofuranosyl)cytosine (8) and 5-amino-1-(2,3-dideoxy-β-D-ribofuranosyl)cytosine (9β). The prepared compounds were tested for their activity against HIV and HBV viruses, but they did not show significant activity.