Pregnancy-specific protein B (PSPB), progesterone and some biochemical attributes concentrations in the fetal fluids and serum and its relationship with fetal and placental characteristics of Iraqi riverine buffalo (Bubalus bubalis).

This study was carried out to demonstrate the pregnancy-specific protein B (PSPB), progesterone and some biochemical parameters concentrations in amniotic fluid, allantoic fluid and fetal serum collected from slaughtered Iraqi riverine pregnant buffaloes at three different months of gestation (6th, 7th and 8th). Ten out of 22 adult buffaloes of 4.6 ± 0.97 years old were used in this study. The buffaloes were mated naturally by monitoring the estrus cycles via appearance of vaginal fluids and mounting by bulls. Pregnancy was checked for these buffaloes by non-returning to estrus for three estrus cycles and assured by rectal palpation on day 61 post-mating (PM). Buffaloes were slaughtered at three different periods of gestation (three at 6th month, four at 7th month and three at 8th month of gestation) to verify the progesterone and PSPB as well as some blood attributes levels (glucose, cholesterol, total protein, albumin, globulins and albumin: globulins ratio) in amniotic fluid (AF), allantoic fluid (LF) and fetal serum (FS). Progesterone was higher (P<0.01) in LF at the 8th month of gestation and lower in FS during the 7th and 8th months of pregnancy. PSPB concentrations were greater in FS (6th and 8th months in particular) than in both AF and LF. The overall mean of cholesterol concentration was higher in FS (P<0.05) followed by AF and LF that had the lowest concentration. The FS exhibited higher total protein during the three gestation periods. Most of fetal and placental measurements increased as the pregnancy advanced. In conclusion, these results described, for the first time, the PSPB and progesterone concentrations and blood characteristics in fetal fluids and serum in water riverine buffaloes during different stages of pregnancy. Progesterone concentrations were greater in allantoic fluid than in other fluids. In contrast, PSPB and other blood attributes were higher in fetal serum than other fluids of Iraqi riverine buffaloes. These findings reflect the changes in hormones, proteins and other metabolites during different gestation periods.

Early pregnancy detection of Iraqi riverine buffalo (Bubalus bubalis) using the BioPRYN enzyme-linked immunosorbent assay for PSPB and the progesterone assay.

This study was undertaken to detect pregnancy in Iraqi riverine buffalo (Bubalus bubalis) using three different methods (rectal palpation, plasma progesterone concentration and detection of the presence of pregnancy-specific protein B (PSPB) with the BioPRYN(®) enzyme-linked immunosorbent assay (ELISA) test. The aim of the study was to identify the most sensitive, early and accurate method for detecting pregnancy. Twenty-two female riverine buffalo that were 6.0 ± 0.93 years old were used. Four blood samples per buffalo were taken via jugular venipuncture at days 22-24, 32-34, 42-44 and 58-61 post-mating (PM) to measure the progesterone concentration (ng/ml) and to detect the presence of plasma PSPB. The rectal palpation method was employed to evaluate all buffalo on days 42-44 and 58-61 PM. The BioPRYN(®) test differed (p<0.01) from the other tests with earlier accuracy for detecting pregnant and non-pregnant buffalo. Eighty-eight percent of pregnant and 76.9% of non-pregnant buffalo were distinguished early (days 22-24 PM) using BioPRYN(®) and plasma PSPB-ELISA level (2.09 ± 0.12 ng/ml) in relation to 66.7% and 53.9% detected using the progesterone assay at similar days (4.30 ± 0.40 ng/ml). In conclusion, these results described, for the first time, the early and accurate pregnancy detection of water riverine buffalo using BioPRYN(®) technology and provided the plasma levels of PSPB using an ELISA test. These findings will improve the reproductive and productive efficiency of Iraqi riverine buffalo by adapting the recent management and reproductive strategies in Iraq and in the world.

Trans-splicing in chloroplasts: the rps 12 loci of Nicotiana tabacum.

The rps12 gene in tobacco chloroplasts consists of three exons that code for a polypeptide with homology to Escherichia coli ribosomal protein S12. C-terminal exons 2 and 3 of rps12 are located in the inverted repeat regions of the tobacco chloroplast genome. Exon 1 of rps12 is 29 kilobase pairs downstream of the nearest copy of exons 2 and 3 and 69 kilobase pairs away from the distal copy of exons 2 and 3. RNA gel blot hybridization analysis and primer extension sequencing of cDNA to rps12 encoding RNAs indicate that exon 1 and exons 2 and 3 are encoded on separate transcripts. Exon 1 and exons 2 and 3 are covalently ligated in the correct reading frame in rps12 mRNA. These results indicate that a bimolecular (trans-) splicing event occurs during the formation of mature rps12 mRNA.

Nucleotide sequence of a cDNA clone carrying the glycoprotein gene of infectious hematopoietic necrosis virus, a fish rhabdovirus.

The nucleotide sequence of the mRNA encoding the glycoprotein of infectious hematopoietic necrosis virus was determined from a cDNA clone containing the entire coding region. The G-protein cDNA is 1,609 nucleotides long (excluding the polyadenylic acid) and encodes a protein of 508 amino acids. The predicted amino acid sequence was compared with that of the glycoprotein of the Indiana and New Jersey serotypes of vesicular stomatitis virus and with the glycoprotein of rabies virus, using a computer program which determined optimal alignment. An amino acid identity of approximately 20% was found between infectious hematopoietic necrosis virus and the two vesicular stomatitis virus serotypes and between infectious hematopoietic necrosis virus and rabies virus. The positions and sizes of the signal sequence and transmembrane domain and the possible glycosylation sites were determined.

Location, nucleotide sequence and expression of the proton-translocating subunit gene of theE. gracilis chloroplast ATP synthase.

A 1700 base pairHindIII restriction fragment from theE. gracilis chloroplast chromosome has been shown to contain the gene for the proton-translocating subunit of the ATP synthase (atpH). The gene was mapped by heterologous hybridization using internal sequences of the gene from wheat and spinach chloroplast DNA. Each chloroplast chromosome contains a single copy of atpH, which is located close to the gene for the alpha subunit of the ATP synthase (atpA). The nucleotide sequence of the gene is uninterrupted by introns. The predicted sequence of 77 amino acids is 70% homologous to that of the wheat and spinach polypeptides. The gene is expressedin vivo as shown by hybridization of atpH probes to cellulose nitrate filter blots of total chloroplast RNA. 5'-proximal to the atpH locus on the clonedHindIII fragment are 38 codons from the carboxy terminus of an open reading frame which may be another ATP synthase subunit. Transcription of the ORF and the atpH gene may be as part of a large polycistronic mRNA.

Location of the single gene for elongation factor Tu on the Euglena gracilis chloroplast chromosome.

The structural gene for elongation factor Tu (EF-Tu) has been mapped by heterologous hybridization to a 2900 base pair sequence of Euglena gracilis Klebs Strain Z Pringsheim chloroplast DNA within the EcoRI fragment, Eco N. The hybridization probes were obtained from a HhaI restriction fragment containing internal sequences to Escherichia coli EF-Tu, located in the tufA gene locus, and from an EcoRI restriction fragment of chloroplast DNA from the eukaryotic algae Chlamydomonas reinhardii, containing the 3' end of the chloroplast EF-Tu gene. This is the second identified protein gene locus to be mapped on the E. gracilis chloroplast genome, and the first using prokaryotic DNA as a probe.

Role of two siderophores in Ustilago sphaerogena. Regulation of biosynthesis and uptake mechanisms.

Under iron-deficient conditions the smut fungus Ustilago sphaerogena produces two kinds of siderophores, ferrichrome and ferrichrome A. Regulation of ligand biosyntheses and uptake mechanisms of the iron chelates were studied to determine the role of each chelate in U. sphaerogena. The biosynthesis of each ligand was differentially regulated. Ferrichrome A, the more effective chelate, was preferentially synthesized under more extreme conditions of iron stress, but completely repressed when the cell was supplied with sufficient iron. In contrast, biosynthesis of ferrichrome was strongly but not completely repressed by iron. The mechanism of repression was examined using a newly developed in vivo synthesis assay. Chromium and gallium-containing siderophore analogs had no effect on siderophore ligand biosynthesis. Iron, added as siderophores, resulted in increased oxygen uptake and amino acid transport, which was soon followed by decreased ligand biosynthesis, suggesting that regulation may be indirect and related to oxidative metabolism. Uptake experiments were used to rule out a ligand-exchange mechanism for ferrichrome A-iron transport. The data suggest that ferrichrome A-iron is taken up at a specific site that results in a rapid distribution of iron inside the cell.