scholarly journals RT-QuIC detection of CWD prion seeding activity in white-tailed deer muscle tissues

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Manci Li ◽  
Marc D. Schwabenlander ◽  
Gage R. Rowden ◽  
Jeremy M. Schefers ◽  
Christopher S. Jennelle ◽  
...  
Keyword(s):  
North America ◽  
Skeletal Muscles ◽  
Economic Losses ◽  
Food Chains ◽  
Wasting Disease ◽  
Additional Protocol ◽  
Muscle Tissues ◽  
Highly Sensitive

AbstractChronic wasting disease (CWD) is a prion disease circulating in wild and farmed cervid populations throughout North America (United States and Canada), Europe (Finland, Norway, Sweden), and South Korea. CWD is a long-term threat to all cervid populations and to cervid hunting heritage, with the potential to cause substantial economic losses across multiple sectors. In North America, hunting and farming industries focused on the processing and consumption of white-tailed deer (WTD) venison are particularly vulnerable to CWD prion contamination, as millions of WTD are consumed annually. Real-time quaking-induced conversion (RT-QuIC) is a highly sensitive assay amplifying misfolded CWD prions in vitro and has facilitated CWD prion detection in a variety of tissues and excreta. To date, no study has comprehensively examined CWD prion content across bulk skeletal muscle tissues harvested from individual CWD infected WTD. Here, we use RT-QuIC to characterize prion-seeding activity in a variety of skeletal muscles from both wild and farmed CWD-positive WTD. We successfully detected CWD prions in muscles commonly used for consumption (e.g., backstrap, tenderloin, etc.) as well as within tongue and neck samples of WTD. Our results suggest that CWD prions are distributed across the skeletal muscles of infected WTD. We posit that RT-QuIC will be a useful tool for monitoring CWD prions in venison and that the method (with additional protocol optimization and high-throughput functionality) could be used to reduce and/or prevent CWD prions from entering animal and human food chains.

scholarly journals RT-QuIC detection of CWD prion seeding activity in white-tailed deer muscle tissues

2021 ◽  
Author(s):  
Manci Li ◽  
Marc D. Schwabenlander ◽  
Gage R. Rowden ◽  
Jeremy M. Schefers ◽  
Michelle Carstensen ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
North America ◽  
Skeletal Muscles ◽  
Economic Losses ◽  
Food Chains ◽  
Sensitive Assay ◽  
Wasting Disease ◽  
Muscle Tissues ◽  
Highly Sensitive

Chronic wasting disease (CWD) is a prion disease circulating in wild and farmed cervid populations throughout North America (United States and Canada), Europe (Finland, Norway, Sweden), and South Korea. CWD is an immediate threat to cervid heritage and the disease is causing substantial economic losses across multiple sectors. In North America, hunting and farming industries focused on the processing and consumption of white-tailed deer (WTD) venison are particularly vulnerable to CWD, as millions of WTD are consumed annually. Real-time quaking-induced conversion (RT-QuIC) is a highly sensitive assay amplifying misfolded CWD prions in vitro and has facilitated CWD prion detection in a variety of tissues and excreta. To date, no study has comprehensively examined CWD prion content across bulk skeletal muscle tissues harvested from individual CWD infected WTD. Here, we use RT-QuIC to quantify prion-seeding activity in a variety of skeletal muscles from both wild and farmed CWD-positive WTD. We successfully detected CWD prions in muscles commonly used for consumption (e.g., backstrap, tenderloin, etc.) as well as within tongue and neck samples of WTD. Our results help to establish the utility of RT-QuIC for monitoring CWD prions in venison and suggest that the method is useful for preventing CWD prions from entering animal and human food chains. Moreover, our work indicates that CWD prions are more widely distributed across skeletal muscles of infected WTD than previously reported.

scholarly journals Human prothymocytes. Membrane properties, differentiation patterns, glucocorticoid sensitivity, and ultrastructural features.

1980 ◽  
Vol 152 (4) ◽  
pp. 796-807 ◽  
Author(s):  
U Galili ◽  
A Polliack ◽  
E Okon ◽  
R Leizerovitz ◽  
H Gamliel ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Mitotic Activity ◽  
Membrane Properties ◽  
Nuclear Chromatin ◽  
Peanut Agglutinin ◽  
Chromatin Pattern ◽  
Highly Sensitive ◽  
Thymic Cell

Thymic precursor cells (prothymocytes) comprise a large proportion of the fetal thymic cell population, but are less frequently encountered in the postnatal thymus, where they compose < 1% of the entire population. In the present study we attempted to characterize a number of properties of the prothymocytes obtained from human fetal thymic tissues after depletion of the E-rosette thymocyes on a Ficoll-Hypaque gradient. The prothymocytes are larger than the thymocytes and show a different nuclear chromatin pattern. This subset of cells lacks the E-rosetting and natural-attachment capacities and, unlike thymocytes, does not bind the lectin peanut agglutinin. Human prothymocytes are highly sensitive to the in vitro cytolytic effect of hydrocortisone, whereas the thymocytes are resistant. Long-term in vitro culture of prothymocytes resulted in the expression of thymocyte characteristics together with a burst of mitotic activity. Results of this study indicate that the rate of the prothymocyte proliferation is regulated by the small thymocytes present in the same suspension.

scholarly journals Antibacterial activity of tannins towards Pseudomonas syringae pv. tomato, and their potential as biostimulants on tomato plants

2021 ◽  
Vol 60 (1) ◽  
pp. 23-36
Author(s):  
Paolo CANZONIERE ◽  
Sara FRANCESCONI ◽  
Samuele GIOVANDO ◽  
Giorgio BALESTRA
Keyword(s):  
Antibacterial Activity ◽  
Water Treatment ◽  
Pseudomonas Syringae ◽  
Crop Protection ◽  
Dry Weight ◽  
Economic Losses ◽  
Copper Hydroxide ◽  
Copper Treatment

Pseudomonas syringae pv. tomato (Pst), the causal agent of bacterial speck of tomato, is a significant cause of economic losses in tomato crops. This disease is mainly controlled with preventive use of cupric salt formulations. Antibacterial activity of the tannins U1, U2, U3 and U4, applied alone at 1% w/v concentration or in combination with half (0.045% w/v) of standard of copper hydroxide treatments, was assayed for effects on Pst. In vitro, the four tannins completely inhibited Pst colony formation after 24 h, but U2 (quebracho tannins) + ½ Cu(OH)2 allowed Pst growth after 48 h of incubation, indicating that, since U2 is composed of high molecular condensed tannins it is likely that their structures have chelated the copper hydroxide much more then hydrolysable ones, thus inactivating copper hydroxide and tannins. In fact, this activity of the tannins was equivalent to that for 0.045% w/v of copper hydroxide. Effects of tannins on tomato plant growth were also assessed. On seedlings, long-term U1 treatments increased dry weight of shoots compared to copper hydroxide, but not to water treatment. The U4 treatment increased the NBI values compared to copper treatment but did not show significant differences compared to the water treatment. Inhibitory activity of tannin treatments reduced disease by 37–62%, and 60% after copper treatment, while disease severity was reduced by 33–54% after treating plants with tannins and 36% after copper treatment. On mature plants treated once, the disease reduction was 27–39% after tannin treatments and 44% after copper treatment, while severity was reduced by 50–60% from tannin treatments, and 47% by copper. In seedlings and mature plants, these reductions were similar (P > 0.05) for the tannins and copper treatments. This study indicates a novel crop protection strategy using natural products as alternatives to xenobiotic compounds.

scholarly journals Innate Immune Responses to Wildtype and Attenuated Sheeppox Virus Mediated Through RIG-1 Sensing in PBMC In-Vitro

2021 ◽  
Vol 12 ◽  
Author(s):  
Tesfaye Rufael Chibssa ◽  
Richard Thiga Kangethe ◽  
Francisco J. Berguido ◽  
Tirumala Bharani K. Settypalli ◽  
Yang Liu ◽  
...  
Keyword(s):  
Immune Responses ◽  
Small Ruminants ◽  
Significant Negative Correlation ◽  
Individual Variability ◽  
Innate Immune ◽  
Economic Losses ◽  
Viral Dsrna ◽  
Sheeppox Virus

Sheeppox (SPP) is a highly contagious disease of small ruminants caused by sheeppox virus (SPPV) and predominantly occurs in Asia and Africa with significant economic losses. SPPV is genetically and immunologically closely related to goatpox virus (GTPV) and lumpy skin disease virus (LSDV), which infect goats and cattle respectively. SPPV live attenuated vaccines (LAVs) are used for vaccination against SPP and goatpox (GTP). Mechanisms related to innate immunity elicited by SPPV are unknown. Although adaptive immunity is responsible for long-term immunity, it is the innate responses that prevent viral invasion and replication before LAVs generate specific long-term protection. We analyzed the relative expression of thirteen selected genes that included pattern recognition receptors (PRRs), Nuclear factor-κβ p65 (NF-κβ), and cytokines to understand better the interaction between SPPV and its host. The transcripts of targeted genes in sheep PBMC incubated with either wild type (WT) or LAV SPPV were analyzed using quantitative PCR. Among PRRs, we observed a significantly higher expression of RIG-1 in PBMC incubated with both WT and LAV, with the former producing the highest expression level. However, there was high inter-individual variability in cytokine transcripts levels among different donors, with the expression of TNFα, IL-15, and IL-10 all significantly higher in both PBMC infected with either WT or LAV compared to control PBMC. Correlation studies revealed a strong significant correlation between RIG-1 and IL-10, between TLR4, TNFα, and NF-κβ, between IL-18 and IL-15, and between NF-κβ and IL-10. There was also a significant negative correlation between RIG-1 and IFNγ, between TLR3 and IL-1 β, and between TLR4 and IL-15 (P< 0.05). This study identified RIG-1 as an important PRR in the signaling pathway of innate immune activation during SPPV infection, possibly through intermediate viral dsRNA. The role of immunomodulatory molecules produced by SPPV capable of inhibiting downstream signaling activation following RIG-1 upregulation is discussed. These findings advance our knowledge of the induction of immune responses by SPPV and will help develop safer and more potent vaccines against SPP and GTP.

scholarly journals Effects of in vitro purging with 4-hydroperoxycyclophosphamide on the hematopoietic and microenvironmental elements of human bone marrow

Blood ◽  
1985 ◽  
Vol 65 (3) ◽  
pp. 655-662
Author(s):  
S Siena ◽  
H Castro-Malaspina ◽  
SC Gulati ◽  
L Lu ◽  
MO Colvin ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Pluripotent Stem Cell ◽  
Cell Concentration ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Hematopoietic Progenitors ◽  
Hematopoietic Stem ◽  
Highly Sensitive

We describe the effects of 4-hydroperoxycyclophosphamide (4-HC) on the hematopoietic and stromal elements of human bone marrow. Marrow cells were exposed to 4-HC and then assayed for mixed (CFU-Mix), erythroid (BFU-E), granulomonocytic (CFU-GM), and marrow fibroblast (CFU-F) colony-forming cells and studied in the long-term marrow culture (LTMC) system. The inhibition of colony formation by 4-HC was dose and cell- concentration dependent. The cell most sensitive to 4-HC was CFU-Mix (ID50 31 mumol/L) followed by BFU-E (ID50 41 mumol/L), CFU-GM (ID50 89 mumol/L), and CFU-F (ID50 235 mumol/L). In LTMC, a dose-related inhibition of CFU-GM production was noted. Marrows treated with 300 mumol/L 4-HC were completely depleted of CFU-GM but were able to generate these progenitors in LTMC. Marrow stromal progenitors giving rise to stromal layers in LTMC, although less sensitive to 4-HC cytotoxicity, were damaged by 4-HC also in a dose-related manner. Marrows treated with 4-HC up to 300 mumol/L, gave rise to stromal layers composed of fibroblasts, endothelial cells, adipocytes, and macrophages. Cocultivation experiments with freshly isolated autologous hematopoietic cells showed that stromal layers derived from 4-HC- treated marrows were capable of sustaining the long-term production of CFU-GM as well as controls. In conclusion: (1) Hematopoietic progenitors cells, CFU-Mix, BFU-E, and CFU-GM, are highly sensitive to 4-HC, whereas marrow stromal progenitor cells are relatively resistant. (2) Marrows treated with 300 mumol/L 4-HC that are depleted of CFU-Mix, BFU-E, and CFU-GM can generate CFU-GM in LTMC, suggesting that most primitive hematopoietic stem cells (not represented by CFU-Mix) are spared by 4-HC up to this dose. (3) Consequently, the above colony assays are not suitable tools for predicting pluripotent stem cell survival after 4-HC treatment in vitro.

scholarly journals Ocean planning for species on the move provides substantial benefits and requires few trade-offs

2020 ◽  
Vol 6 (50) ◽  
pp. eabb8428
Author(s):  
M. L. Pinsky ◽  
L. A. Rogers ◽  
J. W. Morley ◽  
T. L. Frölicher
Keyword(s):  
Climate Change ◽  
North America ◽  
Human Activities ◽  
Opportunity Costs ◽  
Marine Animals ◽  
Trade Offs ◽  
Highly Sensitive ◽  
The Impact ◽  
Additional Area

Societies increasingly use multisector ocean planning as a tool to mitigate conflicts over space in the sea, but such plans can be highly sensitive to species redistribution driven by climate change or other factors. A key uncertainty is whether planning ahead for future species redistributions imposes high opportunity costs and sharp trade-offs against current ocean plans. Here, we use more than 10,000 projections for marine animals around North America to test the impact of climate-driven species redistributions on the ability of ocean plans to meet their goals. We show that planning for redistributions can substantially reduce exposure to risks from climate change with little additional area set aside and with few trade-offs against current ocean plan effectiveness. Networks of management areas are a key strategy. While climate change will severely disrupt many human activities, we find a strong benefit to proactively planning for long-term ocean change.

scholarly journals Plasma and tissue concentrations of α-tocopherol during vitamin E depletion in sheep

1993 ◽  
Vol 69 (1) ◽  
pp. 225-232 ◽  
Author(s):  
J. M. Fry ◽  
G. M. Smith ◽  
M. C Mcgrath ◽  
E. J. Speijers ◽  
J. G. Allen
Keyword(s):  
Adipose Tissue ◽  
Vitamin E ◽  
Heart Muscle ◽  
Skeletal Muscles ◽  
Experimental Conditions ◽  
Deficient Diet ◽  
Tocopherol Concentration ◽  
Muscle Tissues ◽  
The Relationship

To determine the relationship between plasma and tissue α-tocopherol concentrations during vitamin E depletion, weaned lambs were placed on a vitamin E-deficient diet for 0, 1, 2, 4, 8 and 12 weeks. α-Tocopherol was measured in plasma, erythrocytes, liver, adrenal, adipose tissue, three different skeletal muscles and heart muscle. The α-tocopherol concentration in plasma fell at the same rate as the α-tocopherol concentration in skeletal muscles, heart muscle, adrenal and adipose tissue. The α-tocopherol concentration in liver and erythrocytes fell at a faster rate than that of plasma and all muscle tissues. There were significant correlations between α-tocopherol concentration in plasma and α-tocopherol concentrations in all the tissues measured. Different skeletal muscles had significantly different concentrations of α-tocopherol which may relate to their differing susceptibility to nutritional myopathy. The increase in malondialdehyde in oxidatively-stressed muscle tissue and the correlation with α-tocopherol concentration in most muscle tissues indicated that the muscles had reduced antioxidant capacityin vitroas a result of vitamin E depletion. It was concluded that during vitamin E depletion in sheep α-tocopherol concentration in plasma was a good index of vitamin E status under the experimental conditions employed.

scholarly journals Effects of in vitro purging with 4-hydroperoxycyclophosphamide on the hematopoietic and microenvironmental elements of human bone marrow

Blood ◽  
1985 ◽  
Vol 65 (3) ◽  
pp. 655-662 ◽  
Author(s):  
S Siena ◽  
H Castro-Malaspina ◽  
SC Gulati ◽  
L Lu ◽  
MO Colvin ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Pluripotent Stem Cell ◽  
Cell Concentration ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Hematopoietic Progenitors ◽  
Hematopoietic Stem ◽  
Highly Sensitive

Abstract We describe the effects of 4-hydroperoxycyclophosphamide (4-HC) on the hematopoietic and stromal elements of human bone marrow. Marrow cells were exposed to 4-HC and then assayed for mixed (CFU-Mix), erythroid (BFU-E), granulomonocytic (CFU-GM), and marrow fibroblast (CFU-F) colony-forming cells and studied in the long-term marrow culture (LTMC) system. The inhibition of colony formation by 4-HC was dose and cell- concentration dependent. The cell most sensitive to 4-HC was CFU-Mix (ID50 31 mumol/L) followed by BFU-E (ID50 41 mumol/L), CFU-GM (ID50 89 mumol/L), and CFU-F (ID50 235 mumol/L). In LTMC, a dose-related inhibition of CFU-GM production was noted. Marrows treated with 300 mumol/L 4-HC were completely depleted of CFU-GM but were able to generate these progenitors in LTMC. Marrow stromal progenitors giving rise to stromal layers in LTMC, although less sensitive to 4-HC cytotoxicity, were damaged by 4-HC also in a dose-related manner. Marrows treated with 4-HC up to 300 mumol/L, gave rise to stromal layers composed of fibroblasts, endothelial cells, adipocytes, and macrophages. Cocultivation experiments with freshly isolated autologous hematopoietic cells showed that stromal layers derived from 4-HC- treated marrows were capable of sustaining the long-term production of CFU-GM as well as controls. In conclusion: (1) Hematopoietic progenitors cells, CFU-Mix, BFU-E, and CFU-GM, are highly sensitive to 4-HC, whereas marrow stromal progenitor cells are relatively resistant. (2) Marrows treated with 300 mumol/L 4-HC that are depleted of CFU-Mix, BFU-E, and CFU-GM can generate CFU-GM in LTMC, suggesting that most primitive hematopoietic stem cells (not represented by CFU-Mix) are spared by 4-HC up to this dose. (3) Consequently, the above colony assays are not suitable tools for predicting pluripotent stem cell survival after 4-HC treatment in vitro.

Ovarian Dysfunction in Fetally Irradiated Beagles

1977 ◽  
Vol 35 ◽  
pp. 658-659
Author(s):  
T. M. Seed ◽  
M. H. Sanderson ◽  
D. L. Gutzeit ◽  
T. E. Fritz ◽  
D. V. Tolle ◽  
...  
Keyword(s):  
Gamma Rays ◽  
Low Dose ◽  
Long Term Effects ◽  
Ovarian Dysfunction ◽  
Dose Rates ◽  
Highly Sensitive ◽  
Microscopic Evaluation ◽  
Ovarian Pathology ◽  
Normal Offspring

The developing mammalian fetus is thought to be highly sensitive to ionizing radiation. However, dose, dose-rate relationships are not well established, especially the long term effects of protracted, low-dose exposure. A previous report (1) has indicated that bred beagle bitches exposed to daily doses of 5 to 35 R 60Co gamma rays throughout gestation can produce viable, seemingly normal offspring. Puppies irradiated in utero are distinguishable from controls only by their smaller size, dental abnormalities, and, in adulthood, by their inability to bear young.We report here our preliminary microscopic evaluation of ovarian pathology in young pups continuously irradiated throughout gestation at daily (22 h/day) dose rates of either 0.4, 1.0, 2.5, or 5.0 R/day of gamma rays from an attenuated 60Co source. Pups from non-irradiated bitches served as controls. Experimental animals were evaluated clinically and hematologically (control + 5.0 R/day pups) at regular intervals.

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