Linking albinism and immunity: the secrets of secretory lysosomes

Lysosomes are membrane-bound organelles that are found in all mammalian cells and contain hydrolases and lipases required for protein and membrane degradation. In many cells of the immune system, lysosomes also contain secretory proteins that can be released by regulated exocytosis in response to an external stimulus, providing different cell types with a wide range of effector functions. Melanosomes also use a lysosome-related organelle to secrete melanin for pigmentation. Links between albinism and immunity in patients have uncovered a number of key proteins required for lysosomal secretion and have revealed a versatile secretory mechanism that can be fine-tuned by distinct interactions in different cell types.     

Visual discrimination in sharks without optic tectum

After complete removal of the optic tectum, nurse sharks can learn to discriminate black versus white and horizontal versus vertical stripes. This finding is contrary to the traditional belief of exclusive tectal control over visuomotor behavior in lower vertebrates and suggests a role for the telencephalon in the vision of these primitive animals.     

Methods for Characterization of Residual Starch in Distiller's Dried Grains with Solubles (DDGS)

The objective of this study was to establish methods for determining the content and components of residual starch in distiller's dried grains with solubles (DDGS), a coproduct from dry‐grind corn ethanol production. Four DDGS prepared in our laboratory and one DDGS obtained from a commercial ethanol manufacturer were used for the study. Quantitative analysis of total residual sugar (TRS) in DDGS was performed by determining d ‐glucose produced by enzymatic hydrolysis of oligosaccharides and residual starch remaining in hexane‐defatted DDGS after being dispersed in 90% DMSO. The TRS consisted of free glucose, oligosaccharides, and residual starch. The commercial manufacturer's DDGS contained more TRS (15.8%, w/w db) than the laboratory‐processed DDGS (2.4–2.9%, w/w db). The content of residual starch remaining in the commercial DDGS (5.5% w/w db) was also larger than the laboratory‐processed DDGS (1.9–2.5% w/w db). Analyses of molecular weight distribution showed that the residual starch in DDGS consisted of short‐chain amylose and amylopectin, respectively, as the major and minor components. The short‐chain amylose molecules constituted 86.5–94.1% of the residual starch. The major population of the short‐chain amyloses had an average degree of polymerization (DP) of 85, closely resembling the length of enzyme‐resistant fragments of amylose‐lipid complexes.     

Immunomagnetic isolation of canine circulating endothelial and endothelial progenitor cells

Background: Increased concentrations of circulating endothelial cells (CECs) are thought to be a biomarker of vascular injury in human patients with cardiovascular disease, neoplasia, vasculitis, sickle cell anemia, shock, and sepsis. Immunomagnetic isolation is a technique currently used to enumerate human CECs and can detect low numbers of cells. Objectives: The purpose of this study was to determine whether a standard protocol for immunomagnetic isolation could be used to obtain and enumerate CECs and a subpopulation of endothelial progenitor cells (EPCs) from canine whole blood. Methods: Cultured canine aortic endothelial cells were stained immunohistochemically with von Willebrand factor to verify morphology and number. Using magnetic beads conjugated with anti‐CD146, CECs/EPCs were isolated in culture and in canine whole blood. CD146‐positive cells were stained with fluorescein‐conjugated Ulex europaeus agglutinin 1 (UEA‐1) to confirm endothelial origin and cells were counted manually using a fluorescent microscope. The method was then applied to EDTA‐anticoagulated whole blood samples from 10 healthy client‐owned dogs. Results: The anti‐CD146–coated magnetic beads (>5/cell) bound the cultured canine aortic endothelial cells. Only rare UEA‐1–positive cells were obtained from whole blood, while >85–90% of cultured canine aortic endothelial cells were UEA‐1 positive. The percentage recovery of cultured canine aortic endothelial cells was >86%. CECs in canine whole blood had >8 beads attached to the surface and were 10–40 μm in size. Using immunomagnetic isolation, 43.4 ± 15.6 CECs/mL (range 24–70/mL) were isolated from canine whole blood samples. Conclusions: Immunomagnetic isolation is an acceptable method for enumerating canine CECs/EPCs in whole blood. Further studies are warranted to evaluate the clinical significance of CEC/EPC concentration in different canine diseases.     

The influence of age and Rhodococcus equi infection on CD1 expression by equine antigen presenting cells

There is a distinct age-associated susceptibility of horses to Rhodococcus equi infection. Initial infection is thought to occur in the neonatal and perinatal period, and only foals less than 6 months of age are typically affected. R. equi is closely related and structurally similar to Mycobacterium tuberculosis, and causes similar pathologic lesions. Protective immune responses to M. tuberculosis involve classical major histocompatibility complex (MHC)-restricted T cells that recognize peptide antigen, as well as MHC-independent T cells that recognize mycobacterial lipid antigen presented by CD1 molecules. Given the structural similarity between these two pathogens and our previous observations regarding R. equi-specific, MHC-unrestricted cytotoxic T lymphocytes (CTL), we developed 3 related hypotheses: (1) CD1 molecules are expressed on equine antigen presenting cells (APC), (2) CD1 expression on APC is less in foals compared to adults and (3) infection with live virulent R. equi induces up-regulation of CD1 on both adult and perinatal APC. CD1 expression was examined by flow cytometric analysis using a panel of monoclonal CD1 antibodies with different species and isoform specificities.

Results

Three CD1 antibodies specific for CD1b showed consistent cross reactivity with both foal and adult monocyte-derived macrophages (MDM). CD1b and MHC class II expression were significantly higher on adult MDM compared with foals. R. equi infected MDM showed significantly lower expression of CD1b, suggesting that infection with this bacterium induces down-regulation of CD1b on the cell surface. Histograms from dual antibody staining of peripheral blood mononuclear cells also revealed that 45–71% of the monocyte population stained positive for CD1b, and that the majority of these also co-expressed MHC II molecules, indicating that they were APC. The anti-CD1 antibodies showed no binding or minimal binding to bronchoalveolar lavage (BAL)-derived macrophages.

Conclusion

The CD1b isoform is evolutionarily conserved, and is present on equine MDM, as well as on circulating blood monocytes. The unique susceptibility of foals to R. equi infection may be due in part to lower expression of CD1 and MHC class II, as observed in this study. The data also suggests that infection with R. equi induces down-regulation of CD1b on equine MDM. This may represent a novel mechanism by R. equi to avoid detection and killing of infected cells by the immune system, similar to that observed when human APC are infected with M. tuberculosis.     

动物遗传资源:我们为什么要保护以及怎样去保护

一多样性起源 当今我们所提到的动物遗传资源其历史起源于1～1．2万年前新石器时代的农业革命时期,世界上的多个地区分别兴起了农业革命,除了澳洲以外,这场革命遍布了其他所有的有人类居住的大陆。     

Use of a wireless system to measure invasive arterial blood pressure in ponies – preliminary study

Objective  To evaluate the feasibility and functionality of intra-carotid wireless device implantation in ponies, and to investigate its short-term complications.
Study design  Prospective preliminary study.
Animals  Five mixed breed, adult, intact male ponies weighing 104 ± 28.8 kg (mean ± SD) underwent surgery. Arterial blood pressure data were continuously collected from four animals.
Methods  General anesthesia was induced on two consecutive days. On the first day, an intra-arterial wireless device was implanted in the right carotid artery. On the next day, a transcutaneous intra-arterial catheter was placed in the left facial artery. Data from both sources were collected. Post-mortem examination was performed.
Results  Surgical time was 27.1 ± 11.85 minutes. All catheters remained in place with some extra vascular migration. Complications included mild seroma and hematoma.
Conclusion  The wireless system allowed continuous monitoring in ponies throughout anesthesia and at rest and may allow for the recording of arterial blood pressure and heart rate when it would be difficult to achieve with a conventional system (e.g. during recovery from anesthesia).
Clinical relevance  The wireless invasive blood pressure monitor may allow continuous measurements when only intermittent measurements would be feasible with a wired system.     

Fructan and free fructose content of common Australian vegetables and fruit

Fructans are not digested in the small intestines of humans. While many health benefits have been attributed to these carbohydrates, they can cause gastrointestinal symptoms in some individuals. We measured the total fructans in 60 vegetables and 43 fruits using the Megazyme fructan assay. Vegetables with the highest quantity of fructans included garlic, artichoke, shallots, leek bulb, and onions (range, 1.2-17.4 g/100 g fw). Fruits with low, but detectable, fructans included longon, white peach, persimmon, and melon (range, 0.21-0.46 g/100 g fw). The fructan assay was modified to provide an estimate of the average chain length (degree of polymerization) for high fructan vegetables. d-Fructose can also be malabsorbed in the small intestine of humans, so the d-fructose content in some foods was measured to supplement the current food tables. Research in this area will be facilitated through the availability of more comprehensive food composition data.