Marking to weaning production aspects of lambs provided with NSAID analgesia compared with lambs receiving no analgesia at the time of elastrator ring marking.

The provision of analgesia at the time of marking has been adopted by the Australian sheep industry, but data on production benefits are lacking. In the current study, alternate lambs were provided with either meloxicam (non-steroidal anti-inflammatory drug [NSAID], n = 781) or no analgesia (NONE, n = 822) at the time of ring castration and tail docking. Six distinct management groups of lambs were studied. Lambs were weighed immediately before marking and then again at weaning. There was no significant effect of treatment on average daily gain between marking and weaning in cross-bred lambs. In Merino lambs, average daily gain was 5 g/day lower (P < 0.005) in lambs receiving NSAID, but this may not be biologically significant. Lamb losses were significantly (P < 0.05) lower in the NSAID group (1.1%) than in the NONE group (2.7%). This observation is worth validating in larger studies, particularly considering that lamb mortality is a significant cost to production and welfare concern.

Infrared thermal imaging as a method to evaluate heat loss in newborn lambs.

Thermal imaging technology has been identified as a potential method for non-invasive study of thermogenesis in the neonatal lamb. In comparison to measurement of the core body temperature, infrared thermography may observe thermal loss and thermogenesis linked to subcutaneous brown fat depots. This study aimed to identify a suitable method to measure heat loss in the neonatal lamb under a cold challenge. During late pregnancy (day 125), ewes were subjected to either shearing (n=15) or mock handling (sham-shorn for 2min mimicking the shearing movements) (n=15). Previous studies have shown an increase in brown adipose tissue deposition in lambs born to ewes shorn during pregnancy and we hypothesized that the shearing treatment would impact thermoregulatory capacities in newborn lambs. Lambs born to control ewes (n=14; CONTROL) and shorn ewes (n=13; SHORN) were subjected to a cold challenge of 1h duration at 4h after birth. During the cold challenge, thermography images were taken every 10min, from above, at a fixed distance from the dorsal midline. On each image, four fixed-size areas were identified (shoulder, mid loin, hips and rump) and the average and maximum temperatures of each recorded. In all lambs, body surface temperature decreased over time. Overall the SHORN lambs appeared to maintain body surface temperature better than CONTROL lambs, while CONTROL lambs appeared to have higher core temperature. At 30min post cold challenge SHORN lambs tended to have higher body surface temperatures than lambs (P=0.0474). Both average and maximum temperatures were highest at the hips. Average temperature was lowest at the shoulder (P<0.05), while maximum temperatures were lowest at both shoulder and rump (P<0.005). These results indicate that lambs born to shorn ewes maintained their radiated body surface temperature better than CONTROL lambs. In conjunction with core temperature changes under cold challenge, this insight will allow us to understand whether increased body surface temperature contributes to increased overall heat loss or whether increased body surface temperature is indeed a mechanism contributing to maintenance of core body temperature under cold challenge conditions. This study has confirmed the utility of infrared thermography images to capture and identify different levels of thermoregulatory capacity in newborn lambs.

Detection of tubule boundaries based on circular shortest path and polar-transformation of arbitrary shapes.

In studies of germ cell transplantation, counting cells and measuring tubule diameters from different populations using labelled antibodies are important measurement processes. However, it is slow and sanity grinding to do these tasks manually. This paper proposes a way to accelerate these processes using a new image analysis framework based on several novel algorithms: centre points detection of tubules, tubule shape classification, skeleton-based polar-transformation, boundary weighting of polar-transformed image, and circular shortest path smoothing. The framework has been tested on a dataset consisting of 27 images which contain a total of 989 tubules. Experiments show that the detection results of our algorithm are very close to the results obtained manually and the novel approach can achieve a better performance than two existing methods.

A shorter interval between irradiation of recipient testis and germ cell transplantation is detrimental to recovery of fertility in rams.

The objective of the current study was to identify an optimal time period for donor cell transplantation after irradiation in sheep. The testes of recipient rams were treated with a single dose of 15 Gray (Gy) irradiation followed by germ cell transplantation either 3 or 6 weeks later. Transplantation of donor cells at 6 weeks after irradiation resulted in production of donor sperm by all five recipient rams compared with 4 of 11 rams transplanted at 3 weeks. Rams transplanted 3 weeks post-irradiation appeared to show reduced libido and fertility. Two rams produced sperm with low motility (< 20%) and two other rams were azoospermic. More than 1 year after cell transfer, there were heavy infiltrates of CD45-positive cells and more fibrous tissue in 9 of 14 recipient testes (seven rams) that received cells 3 weeks after irradiation. Taken together, these results suggest that the interval between irradiation of recipients and germ cell transplantation affects the success rate of the procedure, with a 6-week interval preferable. The elevated inflammatory/immune reaction may be responsible, at least in part, for the reduced fertility and low libido observed in the rams that received cells 3 weeks post-irradiation.

Expression of hepatocyte low-density lipoprotein receptor-related protein is post-transcriptionally regulated by extracellular matrix.

The low-density lipoprotein receptor-related protein (LRP) is a multifunctional member of the low-density lipoprotein receptor family that has been implicated in a variety of physiologic and pathologic processes. However, little is known about LRP regulation at the molecular level, and the factors that might mediate LRP have not yet been characterized. This is particularly true of hepatocytes, an important site of LRP expression. Hepatocyte gene expression is known to be dependent on extracellular matrix composition, although the effect of extracellular matrix on lipoprotein receptor expression has not yet been investigated. Also, the mechanisms by which the extracellular matrix affects hepatocyte gene expression are not well understood. In this study, we show that hepatocyte LRP expression decreases rapidly at the mRNA, protein, and functional levels on collagen type I, but remains high on an Engelbreth-Holm-Swarm sarcoma matrix-preparation, Matrigel. LRP function was assessed with ligand binding studies and a novel cytotoxicity assay, using Pseudomonas exotoxin A. Investigation of the mechanism of LRP down-regulation revealed a two-fold longer LRP mRNA half-life in hepatocytes cultured on Matrigel relative to collagen. Taken together, these studies reveal that LRP expression in primary hepatocytes is dependent on the extracellular matrix, and that matrix-dependent differences in hepatocyte LRP mRNA expression are primarily due to changes in mRNA stability, indicating for the first time that the expression of LRP is subject to post-transcriptional regulation.

The porcine skeletal muscle ryanodine receptor gene structure coding region 1 to 10614 harbouring 71 exons.

The skeletal muscle ryanodine receptor (RYR1) belongs to a family of calcium release channels that are expressed in different tissues. The RYR1 gene is one of the largest genes characterized, so far, containing a 15253 nucleotide ORF in swine. To study the genomic organization of the porcine skeletal muscle ryanodine receptor gene we have isolated seven genomic fragments spanning 72.7 kb of chromosomal DNA of chromosome 6q12. This region harbours exons 1 to 71 coding for 3538 amino acids (69.6%) of the ryanodine receptor 1.

[Structure and expression of the porcine skeletal muscle ryanodine receptor gene]. / Struktur und Expression des porcinen Skelettmuskel-Ryanodin-Rezeptor-Gens.

The ryanodine receptors (RYR) are a family of intracellular Ca2+ release channels that were first identified in the terminal cistenae of the sarcoplasmic reticulum of the skeletal and cardiac muscle. Mutations within the skeletal muscle isoform were shown to cause malignant hyperthermia in swine and man. We have analysed the genomic structure of the porcine skeletal muscle ryanodine receptor and its expression using chimeric reporter gene constructs consisting of the RYR1 gene promoter and the chloramphenicol acetyltransferase gene after transfection in muscle and non-muscle cells.

Regulation of tissue-specific expression of the skeletal muscle ryanodine receptor gene.

The ryanodine receptors (RYR) are a family of calcium release channels that are expressed in a variety of tissues. Three genes, i. e. ryr1, ryr2, and ryr3, have been identified coding for a skeletal muscle, cardiac muscle, and brain isoform, respectively. Although, the skeletal muscle isoform (RYR1) was shown to be expressed predominantly in skeletal muscle, expression was also detected in the esophagus and brain. To analyze the transcriptional regulation of the RYR1 gene, we have constructed chimeric genes composed of the upstream region of the RYR1 gene and the bacterial chloramphenicol acetyltransferase (CAT) gene and transiently transfected them into primary cultured porcine myoblasts, myotubes, and fibroblasts. A 443-base pair region upstream from the transcription start site was sufficient to direct CAT activity without tissue specificity. Deletion of a 61-base pair fragment from the 5'-end of the promoter resulted in a marked reduction of CAT activity in all three tissue types. A similar reduction of expression was observed when using a construct with the first intron in antisense orientation upstream from the promoter. In contrast, the first intron in sense orientation enhanced expression only in myotubes, while expression was repressed in fibroblasts and myoblasts. Gel retardation analyses showed DNA binding activity in nuclear extracts for two upstream DNA sequence elements. Our data suggest that (i) RYR1 gene expression is regulated by at least two novel transcription factors (designated RYREF-1 and RYREF-2), and (ii) tissue specificity results from a transcriptional repression in nonmuscle cells mediated by the first intron.