Protein profiles of enzymatically isolated rumen epithelium in sheep fed a fibrous diet

Background: The rumen wall plays a major role in efficient transfer of digested nutrients in the rumen to peripheral tissues through the portal venous system. Some of these substrates are metabolised in the epithelium during this process. To identify the specific proteins involved in these processes, we used proteomic technologies. Protein extracts were prepared from ventral rumen tissue of six sheep fed a fibrous diet at 1.5× maintenance energy requirements.Using a newly developed method, we were able to enzymatically isolate the epithelial cells from underlying tissue layers, thus allowing cytosol and membrane fractions to be independently analysed using liquid chromatography tandem mass spectrometry(LC MS/MS).Results: Using our procedure we identified 570 epithelial proteins in the Ovis aries sequence database. Subcellular locations were largely cytosolic(n = 221) and extracellular(n = 85). However, a quarter of the proteins identified were assigned to the plasma membrane or organelle membranes, some of which transport nutrients and metabolites. Of these 91 were transmembrane proteins(TMHMM), 27 had an N-terminal signal peptide(signalP) and TMHMM motif,13 had a glycosylphosphatidylinositol(GPI) anchor and signalP sequence, 67 had beta(β) strands or 17 β strands and a transit peptide sequence, indicating the identified proteins were integral or peripheral membrane proteins. Subunits of the 5 protein complexes involved in mitochondrial cellular energy production were well represented. Structural proteins(15%), proteins involved in the metabolism of lipids and proteins(26%) and those with steroid or cytokine action were a feature of the proteome.Conclusion: Our research has developed a procedure to isolate rumen epithelium proteins from the underlying tissue layers so that they may be profiled using proteomic technologies. The approach improves the number of proteins that can be profiled that are specific to the epithelium of the rumen wall. It provides new insights into the proteins of structural and nutritional importance in the rumen epithelium, that carry out nutrient transport and metabolism, cell growth and signalling.     

Low-temperature synthesis of zintl compounds with a single-source molecular precursor

Thermolysis of the heterobimetallic phosphinidene complex [Sb(PCy)3]2- Li6.6HNMe2 (Cy = C6H11) at 303 to 313 kelvin gives Zintl compounds containing (Sb7)3- anions. The complex thus constitutes a stable molecular single-source precursor to Zintl compounds and provides a potential low-temperature route to photoactive alkali metal antimonates. The new chemical reaction involved, which is driven thermodynamically by the formation of P-P bonds, has implications in the low-temperature synthesis of other technologically important materials (such as gallium arsenide).     

IMPROVEMENT IN YIELD AND QUALITY OF KINNOW (CITRUS DELICIOSA X CITRUS NOBILIS) BY POTASSIUM FERTILIZATION

Citrus, especially Kinnow, fruit yield and quality in Pakistan is not competitive with other countries, which could be mainly attributed to a lack of good nutrient management for citrus orchards. Many of the soils under these orchards have been reported as deficient in potassium (K). Therefore, work was initiated for improving citrus fruit yield, size, and quality through K nutrition management. Experiments were conducted in four districts of Punjab including Faisalabad, Toba Tek Singh, Jhang, and Sargodha and four rates of potash, i.e., 0, 50, 75 and 100 kg dipotassium oxide (K₂O) ha^?1 were applied using sulfate of potash (SOP) along with recommended rates of phosphorus (P) and nitrogen (N). Soils of all the selected sites were clay loam or sandy clay loam and deficient in K. The leaf analysis also showed that all the selected orchards were deficient in K. Application of K increased the fruit yield and quality but K at100 kg K₂O ha^?1 was more effective in increasing the fruit weight and size, and peel thickness than other K rates in all the selected orchards. Juice volume and percentage significantly increased when K was applied at 75 kg K₂O ha^?1 at all sites. Nutrient uptake like K, P, and N enhanced with the increase in K application rate, however, the highest calcium (Ca)+ magnesium (Mg) was recorded at 50 kg K₂O ha^?1. The results indicated that all rates of K application improved the fruit yield and quality and reduced fruit dropping, however, 75 kg K₂O ha^?1 rate was more effective as juice volume and percentage, total soluble solid (TSS)/acid ratio and nutrient uptake showed significant improvement.     

MODULATION IN YIELD AND JUICE QUALITY CHARACTERISTICS OF CITRUS FRUIT FROM TREES SUPPLIED WITH ZINC AND POTASSIUM FOLIARLY

Citrus, especially K innow (Citrus deliciosa × Citrus nobilis), fruit yield and quality in Pakistan is not competitive with that of other countries which could be mainly attributed to the lack of good nutrient management for citrus orchards. The yield losses in this fruit crop occur mainly due to heavy fruit dropping. Experiments to overcome these problems were conducted at four different sites one each in Faisalabad, Toba Tek Singh, Jhang and Sargodha districts of Punjab, Pakistan. The soil and leaf chemical analysis showed severe deficiency of Zn and our pervious results have shown that soil amendment with potassium (K) at 75 K₂O kg ha^?1 improved the citrus fruit yield and quality at all selected sites. In the present experiments, effect of foliar application of Zn and K alone or in combination was studied on nutrient uptake, fruit yield, fruit dropping and juice quality. The fruit trees were pretreated with a selected K level of sulfate of potash (SOP) or muriate of potash (MOP), i.e., 75 kg K₂O ha^?1 along with recommended nitrogen (N) and phosphorus (P) doses. Zinc [Zn, 1% zinc sulfate (ZnSO₄) solution], K [1% potassium sulfate (K₂SO₄) solution] and Zn + K (solution containing 0.5% each of ZnSO₄ and K₂SO₄) were sprayed at the onset of spring and flush of leaves or flowers, fruit formation and at color initiation on fruit. Overall, application of Zn, K or Zn + K was effective in improving the nutrient uptake, yield and quality parameters of citrus fruit at all sites. Fruit dropping was also reduced by the foliar spray of Zn, K or Zn + K but the most promising results were recorded with foliar spray containing both Zn and K.     

Zur Reaktion des Roggens in langjähriger Monokultur bei differenzierter Mineral- und Stallmistdüngung

Veränderungen in der Futterqualität von Phalaris arundinacea wurden in Abhängigkeit vom Schnittzeitpunkt im 1. Aufwuchs und variierter Nachwuchsdauer im 2. Aufwuchs untersucht. Daraus ließen sich die optimalen Nutzungszeitspannen von Phalaris arundinacea bestimmen. Überflutungen beeinflussen ebenfalls die Länge der optimalen Nutzungszeitspannen und die Futterqualität. Bei Phalaris arundinacea stieg der Rohfaser‐ und Ligningehalt sowie der K/N—Quotient mit zuneymendem physiologischen Alter an, wogegen der Rohprotein‐ und Kaliumgehalt sank.     

Homogeneous Linewidths in the Optical Spectrum of a Single Gallium Arsenide Quantum Dot

The homogeneous linewidths in the photoluminescence excitation spectrum of a single, naturally formed gallium arsenide (GaAs) quantum dot have been measured with high spatial and spectral resolution. The energies and linewidths of the homogeneous spectrum provide a new perspective on the dephasing dynamics of the exciton in a quantum-confined, solid-state system. The origins of the linewidths are discussed in terms of the dynamics of the exciton in zero dimensions, in particular, in terms of lifetime broadening through the emission or absorption of phonons and photons.     

Early Development and Putative Primordial Germ Cells Characterization in Dogs

Previously, three distinct populations of putative primordial germ cells (PGCs), namely gonocytes, intermediate cells and pre‐spermatogonia, have been described in the human foetal testis. According to our knowledge, these PGCs have not been studied in any other species. The aim of our study was to identify similar PGC populations in canine embryos. First, we develop a protocol for canine embryo isolation. Following our protocol, 15 canine embryos at 21–25 days of pregnancy were isolated by ovaryhysterectomy surgery. Our data indicate that dramatic changes occur in canine embryo development and PGCs specification between 21 to 25 days of gestation. At that moment, only two PGC populations with distinct morphology can be identified by histological analyses. Cell population 1 presented round nuclei with prominent nucleolus and a high nuclear to cytoplasm ratio, showing gonocyte morphology. Cell population 2 was often localized at the periphery of the testicular cords and presented typical features of PGC. Both germ cell populations were positively immunostained with anti‐human OCT‐4 antibody. However, at day 25, all cells of population 1 reacted positively with OCT‐4, whereas in population 2, fewer cells were positive for this marker. These two PGCs populations present morphological features similar to gonocytes and intermediate cells from human foetal testis. It is expected that a population of pre‐spermatogonia would be observed at later stages of canine foetus development. We also showed that anti‐human OCT‐4 antibody can be useful to identify canine PGC in vivo.     

Correction of sickle cell disease in adult mice by interference with fetal hemoglobin silencing

Persistence of human fetal hemoglobin (HbF, α(2)γ(2)) in adults lessens the severity of sickle cell disease (SCD) and the β-thalassemias. Here, we show that the repressor BCL11A is required in vivo for silencing of γ-globin expression in adult animals, yet dispensable for red cell production. BCL11A serves as a barrier to HbF reactivation by known HbF inducing agents. In a proof-of-principle test of BCL11A as a potential therapeutic target, we demonstrate that inactivation of BCL11A in SCD transgenic mice corrects the hematologic and pathologic defects associated with SCD through high-level pancellular HbF induction. Thus, interference with HbF silencing by manipulation of a single target protein is sufficient to reverse SCD.