Structural basis for tail-anchored membrane protein biogenesis by the Get3-receptor complex

Tail-anchored (TA) proteins are involved in cellular processes including trafficking, degradation, and apoptosis. They contain a C-terminal membrane anchor and are posttranslationally delivered to the endoplasmic reticulum (ER) membrane by the Get3 adenosine triphosphatase interacting with the hetero-oligomeric Get1/2 receptor. We have determined crystal structures of Get3 in complex with the cytosolic domains of Get1 and Get2 in different functional states at 3.0, 3.2, and 4.6 angstrom resolution. The structural data, together with biochemical experiments, show that Get1 and Get2 use adjacent, partially overlapping binding sites and that both can bind simultaneously to Get3. Docking to the Get1/2 complex allows for conformational changes in Get3 that are required for TA protein insertion. These data suggest a molecular mechanism for nucleotide-regulated delivery of TA proteins.     

Lectin‐Binding Pattern in Ovarian Structures of Rats with Experimental Polycystic Ovaries

Numerous experimental models in different species have been developed for the study of polycystic ovarian syndrome. In this study, we used a model of induction of polycystic ovaries (PO) in rats by exposure to constant light to study the distribution and variations of glycosylated residues present in the different ovarian structures. Seven biotinylated lectins were used (Con‐A, WGA, DBA, SBA, PNA, RCA and UEA‐I) on tissue sections, and detection was performed using the streptavidin/peroxidase method. In tissue sections was observed an increase in affinity for Con‐A in the granulosa and theca interna of growing follicles and cysts in animals with PO in relation to the control group. Follicular cysts showed higher affinity for WGA and RCA‐I which growing follicles in the same group, and there was a decrease in affinity for PNA in the cysts in relation to the growth of follicles in both groups. Atretic follicles in both groups showed greater labelling with lectins PNA, SBA and RCA‐I in relation to healthy follicles. It could also be noted that the zona pellucida of cystic follicles lost the affinity for the lectin Con‐A. There was no staining on follicles in any category with the lectins DBA and UEA‐I, although it was staining in the corpus luteum (control group) and in the mesothelium and interstitial glands of both groups with DBA. These observations probably reflect changes in the glycosaminoglycans present in the different ovarian compartments or in the glycosylation of cellular components essential for proper follicular dynamics.     

Liquorice and canine Addison's disease

Abstract

Extract

Hypoadrenocorticism, or Addison's disease, is an uncommon en-docrinopathy of dogs (Feldman and Nelson 2004 Feldman, EC and Nelson, RW. 2004. “Hypoadrenocorticism (Addison's disease)”. In Canine and Feline Endocrinology and Reproduction, 3rd, 394–439. Philadelphia, , USA: WB Saunders.  [Google Scholar]). Long-term management of affected dogs in New Zealand involves treatment with oral fludrocortisone acetate (Florinef tablets; Bristol-Myers Squibb (NZ) Ltd, Auckland, NZ), which is a relatively expensive synthetic adrenocortical steroid. Table salt may be added to the food, in an attempt to reduce the amount of fludrocortisone required. Despite fludrocortisone therapy, many Addisonian dogs remain hyponatraemic and hyperkalaemic and the required dose of fludrocortisone may need to be increased during the first year of treatment. Increasing the dose of fludrocortisone is not only expensive, but may also have undesirable effects through its potent glucocorticoid action.     

Mycobacterium avium subsp. paratuberculosis infection in wildlife on three deer farms with a history of Johne's disease

Abstract

AIM: To determine the prevalence of Mycobacterium avium subsp. paratuberculosis (Map) infection in wildlife, in pastoral landscapes with a recent history of clinical Johne's disease in livestock.

METHODS: A total of 449 wild mammals and birds from three farms in the South Island of New Zealand with recent histories of clinical Johne's disease in their deer herds were trapped and examined for gross pathological changes in the gastrointestinal tract. Additionally, individual mesenteric lymph nodes from 380 mammals, and segments of gastrointestinal tract from 32 birds were excised, homogenised and cultured for viable Map bacilli. The prevalence of Map infection was then calculated for the various species. Faecal samples from those mammals which had culture-positive tissues were further cultured for the presence of Map.

RESULTS: Gross pathological changes were identified in the gastrointestinal tract of four brushtail possums, one cat, six ferrets, 12 hares, six hedgehogs, three rabbits, one stoat, and one paradise shelduck. Infection with Map in the gastrointestinal tract was confirmed in only three of these cases, one each of brushtail possums, hares and hedgehogs. In contrast, Map infection in the absence of gross pathological changes was frequently recorded in enteric tract tissues of mammals and birds. Among mammals, Map infection was recorded in 18/73 (25%) brushtail possums, 4/23 (17%) cats, 15/42 (36%) hedgehogs and 29/113 (26%) rabbits. Among birds, intestinal tract tissue Map infection was recorded in 3/17 (18%) paradise shelducks. Among 64 of the 74 mammals which had Map culture-positive tissues, 38% (n=5) of hedgehogs and 11% (n=3) of rabbits also had culture-positive faecal samples.

CONCLUSIONS: This study is the first to identify that Map infection can be prevalent in wildlife in New Zealand. There was a high prevalence of Map infection among both scavenging and grazing wild animals. Both mammals and birds are capable of harbouring viable Map organisms in their gastrointestinal tract; further, viable Map was excreted into the environment via faeces by hedgehogs and rabbits.

CLINICAL RELEVANCE: Previous studies overseas have postulated a role of wildlife as reservoirs of Map infection and possible vectors of Johne's disease to livestock. Here, brushtail possums, hedgehogs and rabbits and in particular were identified as potential wildlife hosts for Map infection in NewZealand. This suggests that several wildlife species could contribute to the persistence of Map infection within a wildlife/livestock complex, and potentially, perhaps more importantly, to the spread of infection between farms.     

The equivalence of the Calcium‐Acetate‐Lactate and Double‐Lactate extraction methods to assess soil phosphorus fertility

A large variety of extraction methods are used worldwide for the estimation of “plant‐available P” in soils. In Germany, the standard extractants are Calcium‐Acetate‐Lactate (CAL) and Double‐Lactate (DL). Until now there is no validated transformation procedure available and studies on the comparability of both methods have reported conflicting evidence. The uncertainty about the equivalence of CAL‐P and DL‐P hinders a direct comparison of the P fertility status and P fertilizer recommendations across Germany. Based on 136 datasets for soil samples from an interlaboratory comparison program and three P fertilization field trial sites, for which plant‐available P had been determined by both the CAL and DL method, we assessed the comparability of both extraction methods and derived simple and multiple regression equations to transform DL‐P into CAL‐P values. On average, DL extracted 30% more P than CAL. However, this strongly depended on soil pH and carbonate content. A simple linear regression model explained 70% of the variance. However, if simple linear regression models were fitted to pH‐specific samples (pH range 4.5 to 7.0) the R² increased to 0.96. Based on an independent validation dataset (n = 48) we demonstrated that such pH‐specific models were more accurate than models that did not consider pH when transforming DL‐P to CAL‐P values. Multiple regression results showed that out of soil pH, C_org, N_t, and C : N ratio, only soil pH improved the model. The transformation equations in this study provide a step towards an improved comparability of P fertility status assessments of soils across Germany.     

Free Oestrone in Adipose Tissue and its Relation to Androstenone and Skatole in Entire Male Pigs

Relationship between free oestrone and boar taint compounds in adipose tissue were studied in two groups of entire male pigs of different breeds. Group A consisted of 33 entire crossbred male pigs (dam Yorkshire and sire backcross Yorkshire x Wild Boar, generation seven). Group B consisted of 194 entire male pigs of crossbreeds between Swedish Hampshire (H) and Finnish Landrace (L), LH x H, H x LH, LH x LH (dam x sire). The measurements of free oestrone in adipose tissue were performed with a new method developed and validated in our laboratories. The standard curve was linear for concentrations of free oestrone ranging from 0.13 to 5.10 ng/g. The method exhibited parallelism of results between serial dilutions and a mean recovery of 97 +/- 13.7%. Intra-assay variations for samples with concentrations of free oestrone from 0.67 to 2.08 ng/g were from 9.23 to 11.94%. Inter-assay variations for the samples with concentrations of free oestrone from 0.89 to 2.96 ng/g were from 3.78 to 10.11%. The levels of free oestrone in fat from group A were well correlated with fat levels of androstenone (r = 0.66; p < 0.001) and levels of oestrone sulphate in peripheral plasma collected at the same time as the fat (r = 0.74, p < 0.001). The levels of free oestrone in fat from group B were significantly correlated to fat levels of androstenone (r = 0.68, p < 0.001) and skatole (r = 0.29, p < 0.001). In group B, age-related differences in fat levels of free oestrone, androstenone and skatole were studied. Free oestrone and skatole levels increased simultaneously at the age of 22 week (p < 0.05 for both), and androstenone levels increased at the age of 26 week (p < 0.05). It was suggested that the levels of free oestrone in adipose tissue might be used for the evaluation of hormonal status of male pigs as an alternative to plasma levels of testicular hormones. The levels of free oestrone might be involved in the regulation of skatole levels in fat as indicated by both the simultaneous increases in skatole and free oestrone levels in fat and positive correlation between skatole and free oestrone.     

Effect of hCG Stimulation on Plasma Androstenone Concentrations and Cytochrome b5 Levels in Testicular Tissue

The effect of Human chorionic gonadotropin (hCG) stimulation on the concentrations of free and conjugated androstenone in plasma was studied in 34 crossbred entire male pigs (Landrace x Swedish Yorkshire). Seventeen pigs were treated with hCG 4 days prior to slaughter and the remaining pigs were treated with sterile saline and served as controls. Blood samples were taken prior to hCG or saline injection and on the day before slaughter and analysed for concentrations of free and conjugated androstenone. Testicular tissue samples were taken at slaughter and analysed for the levels of cytochrome b5 (cyb5) protein. Here we have demonstrated for the first time that hCG stimulation causes an increase in the plasma levels of both free and conjugated androstenone. Not all animals responded in the same way to hCG treatment regarding levels of free and conjugated androstenone demonstrating that individual animals can have differences in their capacity to produce free and conjugated androstenone. We suggest that hCG treatment is a good way to determine the potential for androstenone conjugation when androstenone synthesis in the testis is high. The levels of cyb5 protein in the testis were slightly related (r=0.41, p < 0.10) to free androstenone levels in the pigs after hCG administration, although levels of cyb5 protein were not affected by hCG treatment.     

Ultrasonographic Characteristics of Ovulatory Follicles and Associated Endocrine Changes in Cyclic Ewes Treated with Medroxyprogesterone Acetate (MAP)-releasing Intravaginal Sponges and Equine Chorionic Gonadotropin (eCG)

The aim of this study was to assess the ultrasonographic characteristics of ovulatory follicles in cyclic Western White Face ewes (December) that had received intravaginal sponges containing medroxyprogesterone acetate (MAP; 60 mg) for 12 days, with or without an injection of 500 IU of equine chorionic gonadotropin (eCG) at sponge removal. We hypothesized that quantitative echotextural attributes of the follicles in ewes treated only with MAP would differ from those in MAP/eCG-treated ewes, reflecting the increased antral follicular growth and secretory function under eCG influence. Digital images of ovulatory follicles obtained at 0 and 24 h after MAP sponge removal and at 24 h before ovulation in the eCG-treated (five ewes, 13 follicles) and control (six ewes, 9 follicles) animals, were subjected to computerized analyses. The mean diameter of ovulatory follicles increased (p < 0.001) 24 h after eCG treatment. The mean pixel intensity and heterogeneity of the follicular antrum (p < 0.001), as well as mean pixel intensity of the follicular wall and perifollicular ovarian stroma (p < 0.05), were greater in eCG-treated animals compared with control ewes 24 h after sponge removal and at 24 h before ovulation. Mean serum concentrations of oestradiol-17beta tended to increase (p = 0.06) 24 h after eCG treatment and the eCG-treated ewes exceeded (p < 0.05) control animals in progesterone concentrations from days 9-15 after ovulation. Our results support the hypothesis that large antral follicles in eCG-treated ewes exhibit distinctive echotextural characteristics. Follicular image attributes in eCG-treated ewes appear to be indicative of the changes in follicular morphology and secretory activity caused by the administration of the exogenous gonadotropin, which has both FSH- and LH-like activities.     

Socioeconomic research in agroforestry: a decade in review

Agroforestry research in academia and government and non-governmental agencies began by focusing on biophysical aspects. As this science represents a series of practices implemented by individual farmers, it requires in-depth social and economic analyses to assess economic feasibility of agroforestry systems, factors influencing the adoption of agroforestry, monitor the relevance and effectiveness of investigations, and guide future research efforts. This study presents literature dealing with socioeconomic issues in agroforestry research from interdisciplinary peer-reviewed journals, book chapters, and gray literature from 1992 to 2002. Over 500 publications were analyzed to determine relative dominance of geographical regions, types of analyses, agroforestry technologies, and socioeconomic issues. Trends were examined over time and by region. Overall, the body of literature exhibits an increasing trend toward regional and analytical diversity over time. Some deficiencies of regional and thematic nature were observed, which included underrepresented temperate regions and technologies such as riparian buffers, as well as only a small number of papers dealing with gender and property rights. These suggest opportunities for future investigations.