Equilibrative nucleoside transporter 3 deficiency perturbs lysosome function and macrophage homeostasis

Lysosomal storage diseases (LSDs) are a group of heterogeneous disorders caused by defects in lysosomal enzymes or transporters, resulting in accumulation of undegraded macromolecules or metabolites. Macrophage numbers are expanded in several LSDs, leading to histiocytosis of unknown pathophysiology. Here, we found that mice lacking the equilibrative nucleoside transporter 3 (ENT3) developed a spontaneous and progressive macrophage-dominated histiocytosis. In the absence of ENT3, defective apoptotic cell clearance led to lysosomal nucleoside buildup, elevated intralysosomal pH, and altered macrophage function. The macrophage accumulation was partly due to increased macrophage colony-stimulating factor and receptor expression and signaling secondary to the lysosomal defects. These studies suggest a cellular and molecular basis for the development of histiocytosis in several human syndromes associated with ENT3 mutations and potentially other LSDs.     

Cardiopulmonary bypass in the cat

OBJECTIVE: To assess the physiologic response to, and acute survival of, cats undergoing cardiopulmonary bypass (CPB) and to evaluate the efficacy of a commercial human pediatric oxygenator system on cats weighing less than 6 kg. STUDY DESIGN: Experimental study. ANIMALS: Six intact male cats METHODS: Cats were placed on cardiopulmonary bypass by cannulating the cranial and caudal vena cavae and the carotid artery. The pediatric CPB circuit was primed with 150 mL of a balanced crystalloid solution. Venous drainage was enhanced by a controlled, vacuum-assist system. A cross-clamp was placed on the ascending aorta and cardiac arrest was induced by antegrade infusion of a cold cardioplegia solution. After 45 minutes of arrest time, the cross-clamp was removed and the cats were weaned off bypass and decannulated. No blood products were administered. Heart rate, mean arterial pressure (MAP), central venous pressure, arterial blood gas, hematocrit (HCT), total plasma protein concentration (TP), serum electrolyte concentrations, and activated clotting time (ACT) were measured at baseline period (BL), during CPB, 60 minutes after CPB (CPB 60) and 90 minutes after CPB (CPB 90). A complete blood count (CBC), blood chemistry profile, and urinalysis were performed at BL, during CPB, and CPB 90. Cats were euthanatized after CPB 90. RESULTS: Cardiopulmonary bypass resulted in a significant (P <.05) decrease in mean HCT (18.0%) and TP (2.3 gm/dL) at CPB 90 when compared to BL (30.5% and 6.0 gm/dL, respectively). The MAP at CPB 90 (54 mm Hg) was decreased from BL (94 mm Hg). The ACT increased from a mean of 124 seconds to > 400 seconds with heparinization and was reversed to 300 seconds with protamine. Mean platelet counts decreased from BL (369,000 /microL) to CPB 90 (94,500 /microL). Mean white blood cell counts decreased from 13,200 /microL at BL to 2,200 /microL at CPB 90. Upon reperfusion, 1 cat fibrillated but was successfully defibrillated. CONCLUSIONS: Cardiopulmonary bypass was performed successfully in 6 cats weighing less than 6 kg. Acute survival to 90 minutes after CPB was achieved in all 6 cats CLINICAL RELEVANCE: The ability to perform CPB in the cat may allow intracardiac repair of various heart defects in this species.     

COMPARISON BETWEEN COMPUTED TOMOGRAPHY,FLUOROSCOPY, AND ULTRASONOGRAPHY FOR GUIDING PERCUTANEOUS INJECTION OF THE CANINE INTERVERTEBRAL DISC

A minimally invasive method for delivering injectable therapeutic agents would be desirable for the treatment of intervertebral disc disease in dogs. The purpose of this study was to compare computed tomography (CT), ultrasonography (US), and fluoroscopy modalities for guiding percutaneous injection into canine intervertebral discs. Intervertebral discs of 14 dog cadavers were injected with a gelified ethanol therapeutic agent. Successful injectate placement and injectate leakage were determined based on necropsy inspection of discs. Injection into the nucleus pulposus was successful in 55 of 78 (71%) of all injected discs. Injections guided using CT and fluoroscopy were significantly more successful than US‐guided injections. Odds of successful injection without leakage were greater for CT vs. US (P = 0.0026) but there was no significant difference between CT and fluoroscopy (P = 0.0620). Injection success rates did not differ among vertebral sites or dog cadavers of varying weights. Forty‐nine (63%) of injection sites had injectate leakage outside the disc and 10 of these involved structures within the vertebral canal. The highest rate of injection success with the least amount of leakage was achieved with CT guidance. Findings indicated that CT, fluoroscopy, and US are feasible modalities for guiding percutaneous injection of a gelified ethanol therapeutic agent into the canine intervertebral disc, with moderate to high success rates for different regions of the spine. However, a moderately high rate of injectate leakage occurred outside of the disc and this should be taken into consideration for future safety and efficacy studies.     

Mortality and fish welfare

Mortality has received insufficient attention as a fish welfare topic. Here, we aim to prompt fish farming stakeholders to discuss fish mortalities in relation to welfare. Mortality in farmed fish populations is due to a variety of biotic and abiotic causes, although it is often difficult to differentiate between underlying and immediate causes of mortality. Most mortality appears to occur during episodes associated with disease outbreaks and critical periods (in development or production). Most causes of mortality can be assumed to be associated with suffering prior to death. As mortality rates in farmed fish populations are suspected to rank amongst the highest in commonly farmed vertebrate species, mortality should be a principal fish welfare issue. Long-term mortality rates can be used as a retrospective welfare performance indicator and short-term mortality rates as an operational welfare indicator. Scrutiny of mortality records and determining causes of death will enable action to be taken to avoid further preventable mortality. The welfare performance of fish farms should only be judged on levels of predictable and preventable mortality. Fish farmers will already be monitoring mortality due to commercial and legal requirements. As profitability in fish farming is directly linked to survival, confronting mortality should ultimately benefit both fish and farmers.     

Zinc fertilization of lentil for grain yield and grain zinc concentration in ten Saskatchewan soils

Fertilization of grain legumes with zinc (Zn) can affect both marketable yield and Zn content of the grain, which is important in addressing human nutritional deficiencies in certain regions of the world. A pot experiment was conducted to determine the response of three different market classes of lentil to Zn fertilization using ten surface soils from Saskatchewan (Canada). The distribution of Zn among labile and stable fractions chemically separated from the soil was also determined in the ten prairie soils and related to the lentil responses observed. The three market classes of lentils (large and small green, small red) were grown without Zn (control), and with 2.5 and 5 kg Zn ha^?1 added as zinc sulfate to each soil prior to planting. Zinc fertilizer application significantly influenced grain yield and was soil dependent. A significant increase in grain yield over the control was observed from application of Zn on some low organic matter, high pH Brown Chernozem soils whereas a decrease in grain yield over control was observed in other soils such as a Black Chernozem of high organic matter content and low (<7) pH. Lack of positive yield response to addition of Zn were related to measured high diethylene triamine pentaacetic acid (DTPA) extractable and plant root simulator (PRS) resin membrane probe Zn, and large amounts of native Zn in exchangeable and iron/manganese (Fe/Mn) oxide bound fractions. Application of Zn fertilizer generally increased the grain concentration of Zn. For example, an increase of ～20% in Zn concentration over control was observed when 5 kg Zn ha^?1 was added to a loamy textured low organic matter Brown Chernozem soil. Overall, small green lentil was more consistent in producing a positive response to Zn fertilizer application on soils with low plant available Zn compared to large green lentil and small red lentil.     

Fetal and adult hematopoietic stem cells give rise to distinct T cell lineages in humans

Although the mammalian immune system is generally thought to develop in a linear fashion, findings in avian and murine species argue instead for the developmentally ordered appearance (or "layering") of distinct hematopoietic stem cells (HSCs) that give rise to distinct lymphocyte lineages at different stages of development. Here we provide evidence of an analogous layered immune system in humans. Our results suggest that fetal and adult T cells are distinct populations that arise from different populations of HSCs that are present at different stages of development. We also provide evidence that the fetal T cell lineage is biased toward immune tolerance. These observations offer a mechanistic explanation for the tolerogenic properties of the developing fetus and for variable degrees of immune responsiveness at birth.     

Digestibility of protein and starch from sorghum (Sorghum bicolor) is linked to biochemical and structural features of grain endosperm

Although a principal source of energy and protein for millions of the world's poorest people, the nutritional value of sorghum (Sorghum bicolor L. Moench) is diminished because of low digestibility of grain protein and starch. To address this problem, we analyzed the properties of two sorghum lines that have a common pedigree but differ in digestibility. Consistent with results based on a ruminal fluid assay, the protein and starch of one line (KS48) was more thoroughly digested than that of the other (KS51) using in vitro assays based on pepsin and α-amylase. The indigestibility of KS51 relative to KS48 was shown to be due to (i) a greater abundance of disulfide-bonded proteins; (ii) presence in KS51 of non-waxy starch and the accompanying granule-bound starch synthase; and (iii) the differing nature of the protein matrix and its interaction with starch. The current findings suggest that each of these factors should be considered in efforts to enhance the nutritional value of sorghum grain.