Molecular analysis of the third component of canine complement (C3) and identification of the mutation responsible for hereditary canine C3 deficiency

Genetically determined deficiency of the third component of complement (C3) in the dog is characterized by a predisposition to recurrent bacterial infections and to type 1 membranoproliferative glomerulonephritis. The current studies were undertaken to characterize the cDNA for wild-type canine C3 and identify the molecular basis for hereditary canine C3 deficiency. Amplification, cloning, and sequence analysis indicated that canine C3 is highly conserved in comparison with human, mouse, and guinea pig C3. Southern blot analysis failed to show any gross deletions or rearrangements of DNA from C3-deficient animals. Northern blot analysis indicated that the livers of these animals contain markedly reduced quantities of a normal length C3 mRNA. The full-length 5.1-kb canine C3 cDNA was amplified in overlapping PCR fragments. Sequence analysis of these fragments has shown a deletion of a cytosine at position 2136 (codon 712), leading to a frameshift that generates a stop codon 11 amino acids downstream. The deletion has been confirmed in genomic DNA, and its inheritance has been demonstrated by allele-specific oligonucleotide hybridization.     

Evaluation of pre- and postprandial growth hormone (GH)-releasing hormone-induced GH response in subjects with persistent body weight normalisation after biliopancreatic diversion

BACKGROUND: Obesity is characterised by growth hormone (GH) abnormalities, including a blunted response to stimulation and a 'paradoxical' increase after meals. The blunted GH release is reversed by a surgical intestinal bypass procedure. However, this does not mean that normal GH dynamics have been restored. The present study assessed whether post-surgical weight reduction in obese patients normalised the modulation of GH release produced by metabolic fuels. SUBJECTS: Ten obese female subjects, aged 23-54 y, were studied before and after biliopancreatic diversion (BPD). All patients, after surgery, had experienced a significant reduction in body weight (mean body mass index (BMI) 25.78 +/- 1.01 kg/m2 vs 44.68 +/- 1.73 kg/m2). Two groups were also studied as controls: Ten normal body weight female subjects and ten patients suffering from anorexia nervosa (AN, mean BMI 17.46 +/- 1.12 kg/m2). MEASUREMENTS: We have studied the GH response to a GH releasing hormone (GHRH) bolus (1 microg/kg i.v., at 13.00 h) before and after a standard meal. RESULTS: In post-BPD subjects, the GH response to GHRH in the fasting state, was clearly augmented in comparison with the pre-BPD values (peak values 18.06 +/- 4.56 vs 3.24 +/- 0.68 microg/L). In post-BPD subjects the postprandial GH response was further augmented in comparison with the fasting test (peak 30.12 +/- 4.99 microg/L, P < 0.05). This pattern was similar to that observed in anorexic patients. CONCLUSION: The surgical procedure restores a normal GH response to GHRH in the fasting state, but the 'paradoxical' GH response after meals remains present, suggesting a persistent GH derangement in such patients, which is not related to body weight per se. The surgical procedure makes obese patients similar to anorexics, in the relationships between metabolic fuels and GH secretion. The persistence of the GH postprandial response to GHRH in post-BPD subjects suggests a role for metabolic fuels in the regulation of somatostatin (SRIF) secretion.     

Restoration of PEX2 peroxisome assembly defects by overexpression of PMP70

OBJECTIVES: To investigate articular cartilage collagen network, thickness of birefringent cartilage zones, and glycosaminoglycan concentration in macroscopically normal looking knee joint cartilage of young beagles subjected to experimental slowly progressive osteoarthritis (OA). METHODS: OA was induced by a tibial 30 degree valgus osteotomy in 15 female beagles at the age of 3 months. Fifteen sisters were controls. Cartilage specimens were collected seven (Group 1) and 18 months (Group 2) postoperatively. Collagen induced optical path difference and cartilage zone thickness measurements were determined from histological sections of articular cartilage with smooth and intact surface by computer assisted quantitative polarised light microscopy. Volume density of cartilage collagen fibrils was determined by image analysis from transmission electron micrographs and content of glycosaminoglycans by quantitative digital densitometry from histological sections. RESULTS: In the superficial zone of the lateral tibial and femoral cartilage, the collagen induced optical path difference (birefringence) decreased by 19 to 71% (p < 0.05) seven months postoperatively. This suggests that severe superficial collagen fibril network deterioration took place, as 18 months postoperatively, macroscopic and microscopic OA was present in many cartilage areas. Thickness of the uncalcified cartilage increased while the superficial zone became thinner in the same sites. In operated dogs, glycosaminoglycan content first increased (Group 1) in the lateral tibial condyle and then decreased (Group 2) (p < 0.05). CONCLUSION: In this OA model, derangement of the superficial zone collagen network was the probable reason for birefringence reduction. This change occurred well before macroscopic OA.     

Mechanical properties and resistance to low temperature degradation of surface nitrided 3Y-TZP

3Y-TZP has been one of the most applied ceramics in the biomedical field, specifically for hip, knee and dental implants, given its high strength, moderate fracture toughness and excellent biocompatibility. However, hydrothermal degradation has meant an important disadvantage, as surface t–m transformation, followed by microcracking, can lead to the premature failure of the implant. In this work, surface nitriding at 1600, 1500 and 1400 °C for 1, 2 and 4 h with N₂ gas and ZrN powder was applied and optimised to avoid hydrothermal degradation. Nitriding at 1600 and 1500 °C produces a harder low-toughness surface, not adequate for structural implants. It is shown that the most favourable condition is nitriding at 1400 °C for 1 h, since the mechanical properties of the original 3Y-TZP are not affected with the advantage of retarding hydrothermal degradation by a factor close to 10.     

The Experience in Designing and Evaluating the High Performance Cluster Netuno

This paper presents the Netuno supercomputer, a large-scale cluster installed at Federal University of Rio de Janeiro in Brazil. A detailed performance evaluation of Netuno is presented, depicting its computational and I/O performance, as well as the results for two real-world applications. Since building a high- performance cluster for running a wide range of applications is a non-trivial task, some lessons learned from assembling and operating this cluster, such as the excellent performance of the OpenMPI library, and the relevance of employing an efficient parallel file system over the traditional NFS system, can be useful knowledge to support the design of new systems. Currently, Netuno is being heavily used to run large scale simulations in the areas of ocean modeling, meteorology, engineering, physics, and geophysics.     

Dynamics of Indigenous Microbial Populations of Butter Head Lettuce Grown in Mulch and on Bare Soil

ABSTRACT:  The native microflora of lettuce cultivated in mulch and on bare soil and its evolution during storage at optimal condition were evaluated. Inner, mid, and outer leaves of the lettuce heads were analyzed separately and the evolution of the microbial populations were fitted to Gompertz and logistic models. The cultivation method (bare soil and mulch) introduced differences in the initial counts, evolution, and tolerance to refrigeration temperatures for some of the microbial populations under study. Most microbial populations from mulch lettuce presented a decline or little growth under refrigerated storage. However, populations from bare soil lettuce presented some growth phase during storage. Lactic acid bacteria from bare soil lettuce presented significant growth after 8 d of storage while LAB from mulch grown lettuce did not. Concurrently with the LAB growth, there was a decline in the coliform counts in bare soil grown lettuce. At the end of storage, the inner and mid leaves of mulch lettuce presented lower counts of psychrotrophic bacteria, LAB, and yeast and molds.