Relationship of tissue dimensions and three captive bolt placements on cadaver heads from mature swine (Sus scrofa domesticus) > 200 kg body weight

Three penetrating captive bolt (PCB) placements were tested on cadaver heads from swine with estimated body weight (BW) >200 kg (sows = 232.9 ± 4.1 kg; boars = 229.3 ± 2.6 kg). The objectives were to determine tissue depth, cross-sectional brain area, visible brain damage (BD), regions of BD, and bolt-brain contact; and determine relationships between external head dimensions and tissue depth at each placement. A Jarvis PAS – Type P 0.25R PCB with a Long Stunning Rod Nosepiece Assembly and 3.5 gr power loads was used at the following placements on heads from 111 sows and 46 boars after storage at 2-4° C for approximately 62 h before treatment: FRONTAL (F) – 3.5 cm superior to the optic orbits at midline, TEMPORAL (T) – at the depression posterior to the lateral canthus of the eye within the plane between the lateral canthus and the base of the ear, or BEHIND EAR (BE) – directly caudal to the pinna of the ear on the same plane as the eyes and targeting the middle of the opposite eye. For sows, the bolt path was in the plane of the brain for 42/42 (100%, 95% CI: 91.6-100.0%) F heads, 39/40 (97.5%, 95% CI: 86.8-99.9%) T heads, and 34/39 (87.5%, 95% CI: 72.6-95.7%) BE heads; for the heads that could reliably be assessed for BD damage was detected in 25/26 (96.2%, 95% CI: 80.4-99.9%) F heads, 24/35 (68.6%, 95% CI: 50.7-83.2%) T heads, and 5/40 (12.5%, 95% CI: 4.2-26.8%) BE heads. For boars, the bolt path was in the plane of the brain for 17/17 (100.0%, 95% CI: 80.5-100.0%) F heads, 18/18 (100.0%, 95% CI: 81.5-100.0%) T heads, and 14/14 (100.0%, 95% CI: 76.8-100.0%) BE heads; damage was detected in 11/12 (91.7%, 95% CI: 61.5-99.8%) F heads, 2/15 (13.3%, 95% CI: 1.7-40.5%) T heads, and 7/14 (50.0%, 95% CI: 23.0-77.0%) BE heads. Tissue depth was reported as mean ± standard error followed by 95% one-sided upper reference limit (URL). For sows, total tissue thickness was different (P < 0.05) between placements (F: 52.7 ± 1.0 mm, URL: 64.1 mm; T: 69.8 ± 1.4 mm, URL: 83.9 mm; BE: 89.3 ± 1.5 mm, URL: 103.4 mm). In boars, total tissue thickness was different (P < 0.05) between placements (F: 41.2 ± 2.1 mm, URL: 56.3 mm; T: 73.2 ± 1.5 mm, URL: 83.4 mm; BE: 90.9 ± 3.5 mm, URL: 113.5 mm). For swine > 200 kg BW, F placement may be more effective than T or BE due to less soft tissue thickness, which may reduce concussive force. The brain was within the plane of bolt travel for 100% of F heads with brain damage for 96.2% and 91.7% of F sow and boar heads, respectively.

Comparison of Salt Exclusion in Muscadine and Interspecific Hybrid Grapes Using a Greenhouse Screening Procedure

Bunch grapes (Euvitis) are classified as moderately salt-tolerant. However, little is known about the salt tolerance of muscadine grapes (Vitis rotundifolia). The objective of this research was to evaluate the salt exclusion capacity of muscadine grapes relative to common bunch grape rootstocks and hybrid winegrapes using a greenhouse screening assay. In two separate experiments, 31 muscadine, six bunch grape rootstocks, and five hybrid winegrape cultivars were irrigated daily with a 25-mm sodium chloride salt solution for a period of 14 d, followed by a destructive harvest to determine sodium (Na) and chloride (Cl) concentrations in root and shoot tissues. Generally, the muscadines studied exhibited a greater range of salt concentration relative to bunch grape rootstocks. Total tissue (shoot and root) salt varied by 250% and 430% across muscadines and by 180% and 190% across bunch grape rootstocks for Na and Cl, respectively. Despite the wider range, muscadine grapes expressed significantly less leaf necrosis than the bunch grape rootstocks. The most effective salt-excluding muscadines, ‘Janebell’, ‘Scuppernong’, ‘Late Fry’, and ‘Eudora’, were not distinguishable from the bunch grape rootstocks [‘Paulsen 1103’ (1103P), ‘Ruggeri 140’ (140Ru), ‘Schwarzmann’, ‘Millardet et de Grasset 101-14’ (101-14 Mgt.), ‘Millardet et de Grasset 420A’ (420A), and ‘Matador’]. Overall, there was no discernable difference between the salt exclusion capacity of muscadine and bunch grapes. The hybrid winegrape ‘Blanc Du Bois’ displayed poor Na and Cl exclusion properties but showed only moderate leaf necrosis symptoms. In both experiments, ‘Blanc Du Bois’ accumulated more than two-fold higher root and shoot concentrations of Na and Cl compared with the best-performing rootstocks (1103P, 140Ru, 101-14 Mgt.), suggesting that ‘Blanc Du Bois’ could benefit from grafting if salinity is a limiting factor.

Fibrinolytic system activation immediately following trauma was quickly and intensely suppressed in a rat model of severe blunt trauma

In severe trauma, excessive fibrinolytic activation is associated with an increase in the transfusion volume and mortality rate. However, in the first several hours after a blunt trauma, changes in fibrinolytic activation, suppression, and activation–suppression balance have not yet been elucidated, which the present study aimed to clarify. Anesthetized 9-week-old male Wistar S/T rats experienced severe blunt trauma while being placed inside the Noble–Collip drum. Rats were randomly divided into four groups of seven. The no-trauma group was not exposed to any trauma; the remaining groups were analysed 0, 60, and 180 min after trauma. Immediately following trauma, total tissue-plasminogen activator (tPA) levels significantly increased in the plasma, and the balance of active tPA and active plasminogen activator inhibitor-1 (PAI-1) significantly tipped toward fibrinolytic activation. After trauma, both tPA and PAI-1 levels increased gradually in various organs and active and total PAI-1 levels increased exponentially in the plasma. Total plasma tPA levels 60 min after trauma returned quickly to levels comparable to those in the no-trauma group. In conclusion, fibrinolytic activation was observed only immediately following trauma. Therefore, immediately after trauma, the fibrinolytic system was activated; however, its activation was quickly and intensely suppressed.

New DXA Diagnostic Indexes of Abdominal Obesity

Aim: Cushing’s syndrome (CS) is associated with weight gain and extreme central, visceral, abdominal obesity which is confirmed with dual-energy X-rays absorptiometric (DXA) diagnostic cut-off point (CP) values of central obesity indexes (COI), determined as an android to gynoid tissue and fat mass ratios. These best differentiate CS from non-CS obese women matched with CS according to their age and BMI. The aim of this study was to determine the CP values of new DXA indexes of central, abdominal obesity as a ratio of android and trunk to legs as well as trunk and legs to total tissue and fat mass that best differentiate CS and matched non-CS obese women in order to confirm central abdominal obesity, and to determine their normal CP values that best differentiate healthy non-obese women from CS and non-CS obese women, and to exclude abdominal obesity completely. Material and Methods: DXA indexes of abdominal obesity, calculated as а ratio of regional body fat and tissue mass compartments android to legs (A/L), trunk to legs (Tr/L), trunk to total (Tr/To) and legs to total (L/To) values were determined among 4 groups. Each group consisted of 18 women: 1st group of CS, 2nd group of obese women (O1) not different according to their age and BMI from CS, 3rd group of obese women (O2) with higher BMI of 35 ± 1.2 kg and a 4th group of non-obese, healthy women (C) with a normal BMI. Diagnostic accuracy (DG) of CP values of DXA indexes of abdominal obesity and indexes of normal body fat distribution (BFD) were determined. Results: A/L, Tr/L, Tr/To, and L/To DXA indexes were significantly different between CS and O1 as well as between non-CS women O2 compared to O1 and C. These indexes had a highly significant correlation among each other and also in relation to their BMI (p < 0.0001). A/L-Tm CP value of 0.3 best differentiated the CS from group O1, with the highest DG of 100 % and an A/L-Fm CP value of 0.26 differentiated them with a DG of 94.44% and sensitivity of 100 %. An A/L-Tn CP value of 0.23 and an A/L-Fn CP value of 0.25 best differentiated CS and C as well as O2 and C for the highest DG of 100 %. Conclusions: DXA indexes A/L, Tr/L, Tr/To and L/To values were significantly different among the four groups. These values correlated significantly among them and with their BMI in non-CS groups, thus confirming a BMI increase association with a more pronounced abdominal BFD. An A/L-Tm CP value of 0.3 and an A/L-Fm CP value of 0.26 were discovered as the best DXA diagnostic indexes of extreme abdominal obesity in CS and these could also be used in discovering abdominal BFD in non-CS obese women with metabolic syndrome (MS). An A/L-Tn CP value of 0.23 and an A/L-Fn CP value of 0.25 were discovered as the best DXA diagnostic indexes of normal BFD which completely excluded abdominal obesity.

Impact of charge patches on tumor disposition and biodistribution of therapeutic antibodies

This study explores the impact of antibody surface charge on tissue distribution into various tissues including tumor. Tumor-bearing mice were dosed intravenously with a mixture of three antibodies engineered to carry negative charge patches, a balanced charge distribution, or positive patches, respectively. Tissue levels were analyzed with a specific LC-MS/MS method. In addition, the antibody mix was administered to non-tumor bearing mice. Muscle and skin interstitial fluid were obtained by centrifugation and analyzed by LC-MS/MS. An in-vitro endothelium model was explored for its feasibility to mimic the observed distribution differences. A balanced charge distribution was optimal in terms of total tumor exposure, while in other tissues negatively charged and balanced charged antibodies gave similar results. In contrast, positive charge patches generally result in increased serum clearance but markedly enhance tumor and organ uptake, leading to higher tissue-to-serum ratios. The uptake and availability in the interstitial space were confirmed by specific assessment of antibody levels in the interstitial fluid of muscle and skin, with similar charge impact as in total tissue. The in vitro model was able to differentiate the transport propensity of this series of antibody variants. In summary, our results show the differential effects of charge patches on an antibody surface on biodistribution and tumor uptake. These insights may help in the design of molecules with biodistribution properties tailored to their purpose and an optimized safety profile.

Adipose Stromal Cell-Secretome Counteracts Profibrotic Signals From IPF Lung Matrices

Introduction: Idiopathic pulmonary fibrosis (IPF) is a fibrotic lung disease characterized by excess deposition and altered structure of extracellular matrix (ECM) in the lungs. The fibrotic ECM is paramount in directing resident cells toward a profibrotic phenotype. Collagens, an important part of the fibrotic ECM, have been shown to be structurally different in IPF. To further understand the disease to develop better treatments, the signals from the ECM that drive fibrosis need to be identified. Adipose tissue-derived stromal cell conditioned medium (ASC-CM) has demonstrated antifibrotic effects in animal studies but has not been tested in human samples yet. In this study, the collagen structural integrity in (fibrotic) lung tissue, its interactions with fibroblasts and effects of ASC-CM treatment hereon were studied.Methods: Native and decellularized lung tissue from patients with IPF and controls were stained for denatured collagen using a collagen hybridizing peptide. Primary lung fibroblasts were seeded into decellularized matrices from IPF and control subjects and cultured for 7 days in the presence or absence of ASC-CM. Reseeded matrices were fixed, stained and analyzed for total tissue deposition and specific protein expression.Results: In both native and decellularized lung tissue, more denatured collagen was observed in IPF tissue compared to control tissue. Upon recellularization with fibroblasts, the presence of denatured collagen was equalized in IPF and control matrices, whereas total ECM was higher in IPF matrices than in the control. Treatment with ASC-CM resulted in less ECM deposition, but did not alter the levels of denatured collagen.Discussion: Our data showed that ASC-CM can inhibit fibrotic ECM-induced profibrotic behavior of fibroblasts. This process was independent of collagen structural integrity. Our findings open up new avenues for ASC-CM to be explored as treatment for IPF.

Upregulation of Local Hepcidin Contributes to Iron Accumulation in Alzheimer’s Disease Brains

Background: Accumulation of iron is a consistent feature of Alzheimer’s disease (AD) brains. The underlying cause, however, remains debatable. Objective: To explore whether local hepcidin synthesized by brain cells contributes to iron accumulation in AD brains. Methods: Brain tissue from the cingulate cortex of 33 cases of AD pre-assigned to Braak stage I-VI, 6 cases of non-dementia, and 15 cases of non-AD dementia were analyzed for transcriptional upregulation of hepcidin by RT-qPCR and RT-PCR. Change in the expression of ferritin, ferroportin (Fpn), microglial activation marker Iba1, IL-6, and TGFβ2 was determined by western blotting. Total tissue iron was determined by colorimetry. Results: Significant transcriptional upregulation of hepcidin was observed in Braak stage III-VI relative to Braak stage I and II, non-AD dementia, and non-dementia samples. Ferritin was increased in Braak stage V, and a significant increase in tissue iron was evident in Braak stage III-VI. The expression of Iba1 and IL-6 was also increased in Braak stage III-VI relative to Braak stage I and II and non-AD dementia samples. Amyloid-β plaques were absent in most Braak stage I and II samples, and present in Braak stage III-VI samples with few exceptions. Conclusion: These observations suggest that upregulation of brain hepcidin is mediated by IL-6, a known transcriptional activator of hepcidin. The consequent downregulation of Fpn on neuronal and other cells results in accumulation of iron in AD brains. The increase in hepcidin is disease-specific, and increases with disease progression, implicating AD-specific pathology in the accumulation of iron.

Impact of Carotenoid Cleaving Enzymes on Lycopene Accumulation in Transgenic Mice

Objectives To evaluate the role of β-carotene oxygenase 1 (BCO1) and BCO2 on lycopene tissue distribution. Methods Three-week old C57BL/6 male and female mice (wild type [WT], Bco1−/−, Bco2−/−, Bco1−/− × Bco2−/− double knock out [DKO]) were divided into groups based on genotype (n = 16 per group split evenly by sex) and fed a powdered AIN 93G control diet for 2 weeks. After this period, mice were gavaged daily for 2 weeks with 1 mg of lycopene dissolved in cottonseed oil. 12 h-fasted mice were then sacrificed, and liver, serum, heart, kidney, intestine, gonadal adipose, prostate, spleen, and testes were harvested. Tissues were preserved in liquid nitrogen and stored at −80 until analyses. We measured lycopene levels in all samples by using high-performance liquid chromatography. Data analyses were performed using two-way ANOVA, followed by the Sidaks test with a statistical significance threshold of P &lt; 0.05. Results Female mice showed higher lycopene levels in the intestine (P &lt; 0.045) and liver (P &lt; 0.007) irrespective of genotype, while male mice had higher lycopene levels in serum (P &lt; 0.004). Intestine, serum, and kidneys exhibited higher lycopene levels in DKO mice compared to all other genotypes (P &lt; .0001), while having higher lycopene levels in testes (P &lt; 0.0001) compared to Bco2−/− and WT mice and adipose (P &lt; 0.005) only in comparison to Bco2−/− mice. DKO exhibited higher lycopene levels in the spleen compared to Bco1−/− mice (P &lt; 0.02). Lycopene levels in the liver (P &lt; 0.0001) were higher in Bco2−/− mice compared to Bco1 −/− and DKO mice, while Bco1−/− mice had lower hepatic lycopene levels compared to all other genotypes. Conclusions Female mice accumulated higher lycopene levels in most tissues compared to males. These results were consistent when data were corrected by total tissue weight. The data suggest the absence of BCO2 favors carotenoid accumulation in many extrahepatic tissues, an effect that is enhanced in the absence of both carotenoid cleaving enzymes. Funding Sources Internal funding, University of Illinois Urbana Champaign.

88 Foundational Investigations of Tissue Dimensions and Their Relation to Captive Bolt application Sites on Cadaver Heads from Mature Swine–A Preliminary Report

The objective of this study was to contrast the soft tissue thickness, cranial thickness, total tissue thickness, and cross-sectional brain area from the common frontal captive bolt placement for the captive bolt euthanasia of swine with the alternative temporal and caudal to pinna placements. One hundred and fifty-seven cadaver heads from sows and boars with estimated body weights greater than 200 kg were collected from a regional slaughter establishment following electrical stunning and assigned to the FRONTAL, TEMPORAL, or CAUDAL to pinna captive bolt placement treatments after cooling at 2–4°C for approximately 64 h. In sows, soft tissue thickness was different (P ˂ 0.0001) between the three placements (FRONTAL: 13.9±1.1 mm, TEMPORAL: 45.93±1.1 mm, CAUDAL TO PINNA: 53.8±1.1 mm), cranial thickness was different (P ˂ 0.0001) between the three placements (FRONTAL: 47.1±1.4 mm, TEMPORAL: 17.6±1.4 mm, CAUDAL TO PINNA: 30.2±1.4 mm), total tissue thickness was different (P &lt; 0.0001) between the three placements (FRONTAL: 61.03±1.4 mm, TEMPORAL: 63.49±1.4 mm, CAUDAL TO PINNA: 84.05±1.4 mm), and cross-sectional brain area was different (P &lt; 0.0001) between the three placements (FRONTAL: 4509.0±238.0 mm2, TEMPORAL: 1964.4±238.0 mm2, CAUDAL TO PINNA: 2767.5±238.0 mm2). In boars, soft tissue thickness was different (P &lt; 0.0001) between the three placements (FRONTAL: 12.9±1.7mm, TEMPORAL: 45.3±1.7 mm, CAUDAL TO PINNA: 54.7±1.7 mm), cranial thickness was different (P = 0.0193) between the FRONTAL and TEMPORAL treatments (FRONTAL: 34.8±3.2mm, TEMPORAL: 22.1±3.2 mm, CAUDAL TO PINNA: 31.7±3.2 mm), total tissue thickness was different (P &lt; 0.0001) between the three placements (FRONTAL: 47.7±3.2 mm, TEMPORAL: 67.4±3.2 mm, CAUDAL TO PINNA: 86.4±3.2mm), and cross-sectional brain area was different (P &lt; 0.0001) between the three placements (FRONTAL: 4031.9±153.2mm2, TEMPORAL: 1241.8±153.2 mm2, CAUDAL TO PINNA: 2467.5±153.2 mm2). Overall, the preliminary data indicated that the FRONTAL placement appears to have the greatest likelihood for successful euthanasia and may present less risk than the alternative TEMPORAL or CAUDAL TO PINNA placements.

Quantitative Imaging Analysis of the Spatial Relationship between Antiretrovirals, RT-SHIV RNA, and Collagen in the Mesenteric Lymph Nodes of Nonhuman Primates

HIV persistence in tissue reservoirs is a major barrier to HIV cure. While antiretrovirals (ARVs) suppress viral replication, antiretroviral therapy (ART) interruption results in rapid rebound viremia that may originate from lymphoid tissues. To understand the relationship between anatomic distribution of ARV exposure and viral expression in lymph nodes, we performed mass spectrometry imaging (MSI) of 6 ARVs, RNAscope in situ hybridization for viral RNA, and immunohistochemistry of collagen in mesenteric lymph nodes from 8 uninfected and 10 reverse transcriptase-simian/human immunodeficiency virus (RT-SHIV) infected rhesus macaque nonhuman primates (NHPs) dosed to steady-state with combination ART. MATLAB-based quantitative imaging analysis was used to evaluate spatial and pharmacologic relationships between these ARVs, viral RNA (both vRNA+ cells and FDC-bound virions) and collagen deposition. Using MSI, 31% of mesenteric lymph node tissue area was not covered by any ARV. Additionally, 28% of FDC-trapped virions and 21% of infected cells were not exposed to any detected ARV. Of the 69% of tissue area that was covered by cumulative ART exposure, nearly 100% of concentrations were greater than in vitro IC50 values; however, 52% of total tissue coverage was from only one ARV, primarily maraviroc. Collagen covered ∼35% of tissue area, but did not influence ARV distribution heterogeneity. Our findings are consistent with our hypothesis that ARV distribution, in addition to total-tissue drug concentration, must be considered when evaluating viral persistence in lymph nodes and other reservoir tissues.