Neutrophil Gelatinase–Associated Lipocalin in Dogs with Naturally Occurring Renal Diseases

Background

Neutrophil gelatinase–associated lipocalin (NGAL) is released from renal tubular cells after injury and serves in humans as a real‐time indicator of active kidney damage, including acute kidney injury (AKI) and chronic kidney disease (CKD). However, NGAL concentrations in dogs with naturally occurring AKI or CKD rarely have been explored in detail.

Hypothesis/Objectives

The goal of this study was to evaluate whether NGAL can serve as a useful biomarker in dogs with naturally occurring renal disease.

Animals

Client‐owned dogs with renal disease (57) and control dogs without any disease (12) were examined.

Methods

Serum NGAL (sNGAL) and urine NGAL (uNGAL) concentrations were measured in each animal by a newly developed ELISA system. Demographic, hematologic, and serum biochemical data were recorded. Survival attributable to AKI and CKD was evaluated at 30 days and 90 days, respectively.

Results

Serum and urine NGAL concentrations in azotemic dogs were significantly higher than in nonazotemic dogs and were highly correlated with serum creatinine concentration (P < .05). Among CKD dogs, death was associated with significantly higher sNGAL and uNGAL concentrations compared with survivors. Receiver‐operating characteristic curve (ROC) analysis showed that sNGAL was better than serum creatinine concentration when predicting clinical outcomes for CKD dogs (P < .05). The best cutoff point for sNGAL was 50.6 ng/mL, which gave a sensitivity and a specificity of 76.9 and 100%, respectively. Furthermore, dogs that had higher concentrations of sNGAL survived for a significantly shorter time.

Conclusion

sNGAL is a useful prognostic marker when evaluating dogs with CKD.     

Gene cloning of porcine adiponectin gene from adipose tissue and construction of its eukaryotic expression vector

To clone adiponectin (ADPN) gene from Shaziling porcine adipocyte and construct its eukaryotic expression vector, total RNA was extracted from subcutaneous fatty tissue. One pair of specific primers was designed by Primer 5.0 software according to the sequence of ADPN gene of porcine available in GenBank. The ADPN gene was amplified by PCR from cDNA and cloned into pMD18‐T vector to construct recombinant clonal vector pMD‐ADPN, sequenced and analysed. A recombinant expression plasmid pPICZaA‐ADPN was constructed by subcloning the cloned ADPN gene into the linearized pPICZaA vector. Then, the plasmid pPICZaA‐ADPN was expressed in Pichia pastoris (GS115) by electrotransformation. Western blot and Bradford analysis were used to determine the target protein induced by methanol. Results showed that the genome size of ADPN was 732 bp and encoded 244 amino acid, the nucleotide sequence of ADPN shared 100% identity with that of porcine available in GenBank. Western blot and Bradford analysis showed that the recombinant ADPN was expressed in GS115 correctly and has certain immune activity. The expression level of ADPN was 28.5 μg/ml. In conclusion, the recombinant ADPN could express in eukaryotic expression vector pPICZaA‐ADPN constructed in this study effectively.     

Apoptosis‐Like Events and In Vitro Fertilization Capacity of Sex‐Sorted Bovine Sperm

This study utilized three staining assays (Annexin V, mitochondrial membrane potential (JC‐1) and TUNEL) for flow cytometric analysis of apoptosis in sex‐sorted sperm from four different bulls (A, B, C and D). Correlations between sperm quality and IVF efficiency were then assessed to determine which assay provided the best prediction of IVF efficiency. The results of the Annexin V assays, as well as measures of viable sperm, early apoptosis, necrotic sperm and mitochondrial membrane potential (?ψm) showed that the sex‐sorted sperm collected from bull A significantly differed from those of the other three bulls (p < 0.05). In addition, the levels of DNA fragmentation in sex‐sorted sperm from bull A were significantly lower than those from bulls B and C (p < 0.05). The percentage of cells reaching the cleavage and blastocyst stages in sex‐sorted sperm from bull A were significantly greater than those from the other bulls (p < 0.05). A significant positive correlation was observed between viable sperm and the percentage of cells at the cleavage or blastocyst stages (p < 0.05). In contrast, a negative correlation was found between early apoptotic sperm and the percentage of cells at the cleavage or blastocyst stages (p < 0.05). In conclusion, these results indicated that the Annexin V assay was the most reliable technique for the prediction of the IVF success of sex‐sorted bovine sperm.     

Bio-amelioration of alkali soils through agroforestry systems in central Indo-Gangetic plains of India

A long-term field study was initiated during 1995 at Central Soil Salinity Research Institute, Regional Research Station, Lucknow(26°47′58′′ N and 80°46′24′′ E) to analyze the effect of agroforestry systems on amelioration of alkali soils. Three agroforestry systems(pastoral, silvipastoral and silvicultural) were compared with the control where no agroforestry system was introduced. Tree-based silvicultural and silvipastoral systems were characterized by tree species Prosopis juliflora and Acacia nilotica along with grass species Leptochloa fusca, Panicum maximum, Trifolium alexandrium and Chloris gayana. Growth of ten-year-old Prosopis juliflora and Acacia nilotica planted in combination with grasses was significantly higher over the silviculture system with the same species. Tree biomass yields of P. juliflora(77.20 t?ha-1) and A. nilotica(63.20 t?ha-1) planted under silvipastoral system were significantly higher than the sole plantation of(64.50 t?ha-1 and 52.75 t?ha-1). Fodder yield under the pastoral system was significantly higher than the silvipastoral system during initial years but it was at par with that of silvipastoral systems after eight years of plantation. The microbial biomass carbon in the soils of silvipastoral systems was significantly higher than in soils under sole plantation of trees and control systems. The Prosopis-based silvipastoral system proved more effective in reducing soil pH, displacing Na+ from the exchange complex, increasing organic carbon and available N, P and K. Improvement in soil physical properties such as bulk density, porosity, soil moisture and infiltration rate was higher in the Prosopis-based silvipastoral system than in the silviculture system or control. On the basis of biomass production and improvement in soil health due to tree + grass systems, silvipastoral agroforestry system could be adopted for sustainable reclamation of highly alkali soils.     

Salix tetradenia Hand.‐Mazz: a new natural plant host of ‘Candidatus phytoplasma’

This article reports Salix tetradenia Hand.‐Mazz as a new host of Candidatus phytoplasma and demonstrates its association with witches' broom disease on S. tetradenia plants. Plants exhibited typical visual symptoms of phytoplasma with virescence, abnormality of flowers and witches' broom, and phytoplasma bodies were observed by transmission electron microscopy. Products of 1.2 kb were amplified by nested PCR using phytoplasma universal primer pairs R16F2n/R16R2, but no amplification products were obtained from symptomless plants. The sequence analysis of three 16S rDNA isolates showed 99.84%, 99.68% and 99.76% identify, respectively, with the homologous gene (nc_005303) of member of ‘Candidatus phytoplasma asteris’ (16SrI) group. Phylogenetic and virtual computer‐simulated restriction fragment length polymorphism analysis of the 16S rRNA, tuf and rp gene sequences confirmed that this phytoplasma clustered in the 16SrI‐B subgroup. These results indicated that the diseased S. tetradenia plants were infected by a phytoplasma of the 16SrI group. This is the first report on the occurrence of phytoplasma disease on S. tetradenia worldwide.     

<Emphasis Type="Italic"> Xenostrobus securis </Emphasis>(Lamarck, 1819) (Mollusca: Bivalvia): first report of an introduced species in Galician waters

The presence of the non-indigenous species, the black-pygmy mussel Xenostrobus securis, is reported here for the first time in an intense shellfish farming area off Galicia (NW Spain). Very high concentrations of this mytilid bivalve have colonized estuarine waters located at the inner part of the Ria de Vigo. The invasive role of X. securis is discussed in the context of the wide ecological tolerance of the species and the recent finding of settlements of this species on numerous colonies of the economically-important blue mussel Mytilus galloprovincialis. The mode of introduction of the black-pygmy mussel is also discussed in relation to human management activities.     

Salmon gonadotropes secrete free form alpha (α)-subunit: quantitative analysis by the sandwich cell immunoblot assay

The sandwich cell immunoblot assay confirmed that salmon LH-cells secreted the free form α-subunit along with the dimeric LH, but lower in amounts than the LH.     

Cloning and seasonal changes in ovarian expression of a TSH receptor in the channel catfish, Ictalurus punctatus

The cDNA encoding the thyroid stimulating hormone (TSH) receptor was isolated from the ventral aorta (thyroid follicles) and in both gonads of the channel catfish. The seasonal changes in ovarian expression of this receptor seem to be correlated to reproduction. TSH, but not LH or FSH, activated the recombinant TSH receptor expressed in COS cells. These results indicate that the isolated cDNA encodes a functional and highly specific TSH receptor. The seasonally regulated expression in the catfish ovary suggests a role for TSH in gametegenesis.     

Optimal dietary protein requirement of grouper Epinephelus coioides juveniles fed isoenergetic diets in floating net cages

An experiment to determine the optimal protein requirement of grouper Epinephelus coioides juveniles was conducted in floating net cages (1.5 m × 1 m × 1.5 m). Six isoenergetic fishmeal–casein‐based experimental diets containing 350–600 g kg^?1 crude protein (CP) were fed to triplicate groups of 20 fish (10.7 ± 0.2 g) for 56 days. Weight gain (WG) and specific growth rate (SGR) increased with increasing dietary protein level from 350 to 450 g kg^?1 and then plateaued above these levels. Feed intake (FI) showed no significant difference among fish fed more than 350 g kg^?1 CP. Lowest feed conversion ratio (FCR) was found for fish fed 500 g kg^?1 CP but this was not significantly different from that of fish fed the 450 and 600 g kg^?1 CP. Lowest protein efficiency ratio (PER) was found for fish fed 550 and 600 g kg^?1 CP. Fish fed the 600 g kg^?1 CP had the highest body protein and moisture contents but the lowest body lipid content. Body ash content was unaffected by protein level for fish fed >400 g kg^?1 CP. Dietary protein level had no significant effect on hepatosomatic index (HSI). Fish fed the 350 g kg^?1 CP had significantly lower condition factor (CF) and viscerosomatic index (VSI). Based on broken‐line regression analysis of SGR the optimal dietary protein requirement for E. coioides juveniles was determined to be close to 480 g kg^?1.     

Deduced primary structures of three types of vitellogenin in mosquitofish (Gambusia affinis), a viviparous fish

Three distinct forms of vitellogenin (Vg), 600 kDa VgA and VgB and 400 kDa Vg, were discovered biochemically in estrogen treated female plasma. By sequencing of the three Vg cDNAs, the VgA and VgB were recognized as complete Vgs having all yolk protein (YP) domains, and the 400 kDa Vg was thought to be phosvitinless (Pvl) Vg lacking phosvitin (Pv) domain.