Background Poor nutrition during fetal development programs postnatal kidney function. Understanding postnatal consequences in nonhuman primates (NHP) is important for translation to our understanding the impact on human kidney function and disease risk. We hypothesized that intrauterine growth restriction (IUGR) in NHP persists postnatally, with potential molecular mechanisms revealed by Western-type diet challenge. Methods IUGR juvenile baboons were fed a 7-week Western diet, with kidney biopsies, blood, and urine collected before and after challenge. Transcriptomics and metabolomics were used to analyze biosamples. Results Pre-challenge IUGR kidney transcriptome and urine metabolome differed from controls. Post-challenge, sex and diet-specific responses in urine metabolite and renal signaling pathways were observed. Dysregulated mTOR signaling persisted postnatally in female pre-challenge. Post-challenge IUGR male response showed uncoordinated signaling suggesting proximal tubule injury. Conclusion Fetal undernutrition impacts juvenile offspring kidneys at the molecular level suggesting early-onset blood pressure dysregulation.

Blood pressure (BP) is influenced by genetic variation and sodium intake with sex-specific differences; however, studies to identify renal molecular mechanisms underlying the influence of sodium intake on BP in nonhuman primates (NHP) have focused on males. To address the gap in our understanding of molecular mechanisms regulating BP in female primates, we studied sodium-naïve female baboons (n = 7) fed a high-sodium (HS) diet for 6 wk. We hypothesized that in female baboons variation in renal transcriptional networks correlates with variation in BP response to a high-sodium diet. BP was continuously measured for 64-h periods throughout the study by implantable telemetry devices. Sodium intake, blood samples for clinical chemistries, and ultrasound-guided kidney biopsies were collected before and after the HS diet for RNA-Seq and bioinformatic analyses. We found that on the LS diet but not the HS diet, sodium intake and serum 17 β-estradiol concentration correlated with BP. Furthermore, kidney transcriptomes differed by diet-unbiased weighted gene coexpression network analysis revealed modules of genes correlated with BP on the HS diet but not the LS diet. Our results showed variation in BP on the HS diet correlated with variation in novel kidney gene networks regulated by ESR1 and MYC; i.e., these regulators have not been associated with BP regulation in male humans or rodents. Validation of the mechanisms underlying regulation of BP-associated gene networks in female NHP will inform better therapies toward greater precision medicine for women.

Objective: To evaluate the efficacy of cortical responsive neurostimulation (CRN) in a male baboon with epilepsy and with genetic generalized epilepsy (GGE), as well as the alteration of seizure patterns and their circadian rhythms due to treatment. Methods: The baboon was implanted with two subdural frontoparietal strips, bridging the medial central sulci bilaterally. Electrocorticography (ECoG) data were downloaded daily during a three-month baseline, then every 2-3 days over a five-month treatment period. Long episodes, reflecting ictal or interictal epileptic discharges, were also quantified. Results: Twenty-three generalized tonic-clonic seizures (GTCS) and 2 episodes of nonconvulsive status epilepticus (NCSE) were recorded at baseline (median 8 events/month), whereas 26 GTCS were recorded under treatment (median 5/month). Similarly, daily indices of long episodes decreased from 0.46 at baseline to 0.29 with treatment. Ictal ECoG patterns and the circadian distribution of GTCS were also altered by RNS therapy. Significance: This case study provides the proof-of-concept for RNS therapy in the baboon model of GGE. Cortical responsive neurostimulation (CRN) demonstrated a 38% median reduction in GTCS. Distinct ictal patterns were identified, which changed over the treatment period; the circadian pattern of his GTCS also shifted gradually from night to daytime with treatment. Future studies targeting the thalamic nuclei, or combining cortical and subcortical sites, may further improve detection and control of GTCS as well as other generalized seizure types. More broadly, this study demonstrates opportunities for evaluating seizure detection as well as chronic therapeutic interventions over long term in the baboon. (c) 2021 Elsevier Inc. All rights reserved.

Hypertension is a complex disease influenced by sex, and genetic and environmental factors. Blood pressure (BP) is a continuous trait that is heritable in primates, including humans and baboons. The kidneys play a role in systemically regulating BP. Sex differences in BP onset and control with antihypertensive drug therapies have been observed in humans and rodents. Hypertension studies in nonhuman primates (NHP) to date have focused on males. We hypothesized that there are differences in renal molecular networks associated with BP in female and male primates. Sodium-naïve female (n=8) and male (n=9) baboons were fed a low-sodium chow diet prior to and during the study. Implantable telemetry devices continuously monitored heart rate and blood pressure over 24-hours, and ultrasound-guided kidney biopsies were collected for RNA-Seq. Serum 17 beta-estradiol concentration correlated BP in females. BP in males correlated with Na+ intake, blood urea nitrogen, and glucose. Cell type composition of renal biopsies was consistent between females and males. Sex differences were observed in the kidney transcriptomes by principal components analysis and weighted gene co-expression network analysis. Network analysis revealed HNF4A, ESR1, ESR2, SMARCA4, TP53, and NR3C1 as BP regulators in males. Our results demonstrate sex differences in primate kidney molecular networks and provide evidence of a novel link between renal transcription factors and BP regulation in males. Understanding sex differences and transcriptome variation in primate kidneys correlated with BP and clinical measures associated with BP will inform better therapies towards the goal of precision medicine for women and men.

Background To study central hypovolemia in humans, lower body negative pressure (LBNP) is a recognized alternative to blood removal (HEM). While LBNP mimics the cardiovascular responses of HEM in baboons, similarities in hemostatic responses to LBNP and HEM remain unknown in this species. Methods Thirteen anesthetized baboons were exposed to progressive hypovolemia by HEM and, four weeks later, by LBNP. Hemostatic activity was evaluated by plasma markers, thromboelastography (TEG), flow cytometry, and platelet aggregometry at baseline (BL), during and after hypovolemia. Results BL values were indistinguishable for most parameters although platelet count, maximal clot strength (MA), protein C, thrombin anti-thrombin complex (TAT), thrombin activatable fibrinolysis inhibitor (TAFI) activity significantly differed between HEM and LBNP. Central hypovolemia induced by either method activated coagulation; TEG R-time decreased and MA increased during and after hypovolemia compared to BL. Platelets displayed activation by flow cytometry; platelet count and functional aggregometry were unchanged. TAFI activity and protein, Factors V and VIII, vWF, Proteins C and S all demonstrated hemodilution during HEM and hemoconcentration during LBNP, whereas tissue plasminogen activator (tPA), plasmin/anti-plasmin complex, and plasminogen activator inhibitor-1 did not. Fibrinolysis (TEG LY30) was unchanged by either method; however, at BL, fibrinolysis varied greatly. Post-hoc analysis separated baboons into low-lysis (LY30 <2%) or high-lysis (LY30 >2%) whose fibrinolytic state matched at both HEM and LBNP BL. In high-lysis, BL tPA and LY30 correlated strongly (r = 0.95;P<0.001), but this was absent in low-lysis. In low-lysis, BL TAFI activity and tPA correlated (r = 0.88;P<0.050), but this was absent in high-lysis. Conclusions Central hypovolemia induced by either LBNP or HEM resulted in activation of coagulation; thus, LBNP is an adjunct to study hemorrhage-induced pro-coagulation in baboons. Furthermore, this study revealed a subset of baboons with baseline hyperfibrinolysis, which was strongly coupled to tPA and uncoupled from TAFI activity.

Background Poor nutrition during development programs kidney function. No studies on postnatal consequences of decreased perinatal nutrition exist in nonhuman primates (NHP) for translation to human renal disease. Our baboon model of moderate maternal nutrient restriction (MNR) produces intrauterine growth restricted (IUGR) and programs renal fetal phenotype. We hypothesized that the IUGR phenotype persists postnatally, influencing responses to a high-fat, high-carbohydrate, high-salt (HFCS) diet. Methods Pregnant baboons ate chow (Control; CON) or 70% of control intake (MNR) from 0.16 gestation through lactation. MNR offspring were IUGR at birth. At weaning, all offspring (CON and IUGR females and males, n=3/group) ate chow. At ~4.5 years of age, blood, urine, and kidney biopsies were collected before and after a 7-week HFCS diet challenge. Kidney function, unbiased kidney gene expression, and untargeted urine metabolomics were evaluated. Results IUGR female and male kidney transcriptome and urine metabolome differed from CON at 3.5 years, prior to HFCS. After the challenge, we observed sex-specific and fetal exposure-specific responses in urine creatinine, urine metabolites, and renal signaling pathways. Conclusions We previously showed mTOR signaling dysregulation in IUGR fetal kidneys. Before HFCS, gene expression analysis indicated that dysregulation persists postnatally in IUGR females. IUGR male offspring response to HFCS showed uncoordinated signaling pathway responses suggestive of proximal tubule injury. To our knowledge, this is the first study comparing CON and IUGR postnatal juvenile NHP and the impact of fetal and postnatal life caloric mismatch. Perinatal history needs to be taken into account when assessing renal disease risk.

BACKGROUNDHypotensive resuscitation to raise systolic pressure to 80–90 mm Hg is the standard of care on the battlefield for hemorrhaging Soldiers, and is maintained until the casualty is transported to a surgical unit usually within one hour of injury. Prompt evacuations may not be possible in future conflicts, and hypotension will be required for longer periods of time. This study determined the long‐term effects of prolonged hypotension (PH) held for 0, 1, 2 and 3 hours on kidney, liver and muscle damage up to 21 days after PH, and tested the hypothesis that longer duration of hypotension after hemorrhage will result in greater organ damage.METHODSMale baboons were anesthetized and hemorrhaged until systolic blood pressure was 70 mmHg for 10 min. Systolic blood pressure of greater than 80 mm Hg with autologous blood was maintained for 1 hour (1hr PH; n=5), 2 hours (2hr PH; n=5), or 3 hours (3hr PH; n=5) hours. After the PH period, the hemorrhage volume of whole blood was replaced, and the animal was recovered and monitored for 21 days. Control animals were anesthetized, hemorrhaged and immediately resuscitated with hemorrhaged blood (0hr PH, n=3). Blood samples were obtained before and 1, 3, 7, 14 and 21 days after PH, and analyzed for blood chemistry profile. Data were analyzed with 2 way repeated measures ANOVA and Holm‐Sidak test, and expressed as Mean ± Standard Deviation.RESULTSPlasma creatinine and blood urea nitrogen were unaffected by PH and remained within normal limits in all PH groups. Alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatine phosphokinase (CPK), and lactate dehydrogenase (LDH) values were significantly elevated after PH (time effect; P<0.001; PH group effect; P=0.3). Maximum ALT levels were observed on Day 1 after PH in 0hr PH (140 ± 56 U/L), 1hr PH (170 ± 130 U/L), 2hr PH (322 ± 241 U/L), and 3hr PH (387 ± 167 U/L). Maximum AST levels were observed on Day 1 after PH in 0hr PH (218 ± 44 U/L), 1hr PH (354 ± 219 U/L), 2hr PH (515 ± 424 U/L), and 3hr PH (711 ± 278U/L). Maximum CPK values were observed on Day 1 after PH in 0hr PH (7834 ± 3681 U/L), 1hr PH (24336 ± 22268 U/L), 2hr PH (50494 ± 67653 U/L), and 3hr PH (59857 ± 32408 U/L). Maximum LDH values were observed on Day 1 after PH in 0hr PH (890 ± 396 U/L), 1hr PH (2055 ± 1520 U/L), 2hr PH (3992 ± 4895 U/L), and 3hr PH (4771 ± 1884 U/L). Max levels of ALT, AST, CPK, and LDH were positively correlated with the duration of PH (P<0.05). All values returned to pre‐PH levels by 21 days after PH.CONCLUSIONSThese results indicate that PH up to 3 hours in duration results in transient liver and muscle damage that is correlated with the duration of PH in male baboons. PH of any duration produced minimal effects on the kidney. More studies are needed to determine if longer periods of PH result in permanent and more extensive organ damage.Support or Funding InformationThis study was funded by the United States Army, Medical Research and Materiel CommandThis abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Background: Lower body negative pressure (LBNP) has been established in human studies as an alternative method to blood removal (HEM) to study central hypovolemia. In a baboon model, LBNP also mimics the cardiovascular response of HEM. Hemostatic responses, including platelet activation, coagulation and fibrinolysis in response to LBNP and HEM in this baboon model are reported here. A previous meeting report from this study focused on hemostatic interactions; this report expands the assay of fibrinolytic proteins and includes detailed post-hoc analysis of interactions. Methods: Thirteen anesthetized baboons were exposed to progressive central hypovolemia by HEM and, four weeks later, by LBNP. Whole blood cell counts were determined; pro-coagulant and fibrinolytic activity was evaluated by plasma markers, thrombelastography, flow cytometry and platelet aggregometry at baseline (BL), intermediate hypovolemia (MID), presyncope or maximal hypovolemia (MAX) and after recovery (REC). Results: Baseline values for HEM (day 0) and LBNP (day 28) were indistinguishable for most analyzed parameters; however platelet number, maximal clotting (MA), Protein C, TAT, TAFI-activity, plasma renin, angiotensin and epinephrine were all significantly different (p<0.05). Central hypovolemia induced by both methods activated coagulation; TEG revealed decreased clotting time (R) and increased maximal clotting (MA) at MID, MAX and REC over BL. No differences during developing central hypovolemia were observed between HEM and LBNP. Platelets displayed activation markers on flow cytometry; absolute numbers and functional aggregometry were unchanged. At MAX hypovolemia, there was a modest hemodilution for HEM and a modest hemoconcentration for LBNP; red blood cell count decreased 5.8%±5.5%, p=0.0007 with HEM and increased 5.3±5.9%, p=0.0056 with LBNP. Coagulation factors were significantly increased with LBNP at MAX compared to HEM; fibrinolysis proteins were more varied. No hyper-fibrinolysis developed with progressive hypovolemia by either method; instead hyper-fibrinolysis already existed at BL with significant variance in TEG fibrinolysis at 30 min (LY30) values. Post hoc analysis separated study subjects into low lysis (LY30BL<2) or high lysis (LY30BL>2) whose fibrinolytic state matched at both HEM and LBNP BL. High lysis samples (n=8) showed a strong correlation (r=0.95; p<0.001) of tissue plasminogen activator (tPA) to LY30 at BL that was absent in low lysis samples (n=6). Thrombin activatable fibrinolysis inhibitor (TAFI) activity correlated with tPA levels for low lysis subjects (r=0.88) with no correlation for high lysis subjects (r=.08). Conclusions: Central hypovolemia resulted in activation of coagulation by either LBNP or HEM; thus LBNP is a powerful model for studying hemorrhage. This study also revealed a subset of baboons with baseline activation of fibrinolysis which was strongly coupled to tPA and uncoupled from TAFI activity. The methods demonstrated hemodilution with blood removal and hemoconcentration with LBNP, as has been observed in humans, but these modest changes had no significant effect on coagulation parameters. Unexpectedly, this study demonstrated that the effects of blood removal were sustained 28 days later on baseline parameters. Overall, these results in large primates suggest that baseline differences between apparently healthy and superficially homogeneous individuals may determine outcomes in studies of hemostasis. Disclosures No relevant conflicts of interest to declare.

BACKGROUND This study determined the long-term effects of prolonged hypotension (PH) on liver, muscle, and kidney dysfunction. The hypothesis was that longer duration of PH after hemorrhage will result in greater organ dysfunction. METHODS Baboons were sedated and hemorrhaged (30% blood volume). Systolic blood pressure greater than 80 mm Hg was maintained for 1 hour (1 hr-PH; n = 5), 2 hours (2 hr-PH; n = 5), or 3 hours (3 hr-PH; n = 5). After PH, hemorrhage volume was replaced. Animals were recovered and monitored for 21 days. Control animals were hemorrhaged and immediately resuscitated (0 hr-PH, n = 3). Data are Mean ± Standard Deviation, and analyzed by 2-way repeated measures ANOVA and Holm-Sidak test. RESULTS Hemorrhage resulted in mild hypotension. Minimal resuscitation was required during the hypotensive phase, and survival rate was 100%. Significant increases (p < 0.001) in alanine aminotransferase, aspartate aminotransferase, creatine phosphokinase, and lactate dehydrogenase occurred on Day 1 after PH, and were significantly greater (p < 0.001) in the 2 hr- and 3 hr-PH groups than the 0 hr-PH group. Maximum alanine aminotransferase levels (U/L) were 140 ± 56 (0 hr-PH), 170 ± 130 (1 hr-PH), 322 ± 241 (2 hr-PH), and 387 ± 167 (3 hr-PH). Maximum aspartate aminotransferase levels (U/L) were 218 ± 44 (0 hr-PH), 354 ± 219 (1 hr-PH), 515 ± 424 (2 hr-PH), and 711 ± 278 (3 hr-PH). Maximum creatine phosphokinase values (U/L) were 7834 ± 3681 (0 hr-PH), 24336 ± 22268 (1 hr-PH), 50494 ± 67653 (2 hr-PH), and 59857 ± 32408 (3 hr-PH). Maximum lactic acid dehydrogenase values (U/L) were 890 ± 396 (0 hr-PH), 2055 ± 1520 (1 hr-PH), 3992 ± 4895 (2 hr-PH), and 4771 ± 1884 (3 hr-PH). Plasma creatinine and blood urea nitrogen were unaffected by PH (p > 0.10). CONCLUSION These results indicate that PH up to 3 hours in duration results in transient liver and muscle dysfunction that was most severe after 2 hr-PH and 3 hr-PH. Prolonged hypotension produced minimal effects on the kidney. LEVEL OF EVIDENCE Basic science research, Level of evidence not required for basic science research.

The epileptic baboon provides a natural model of idiopathic generalized epilepsy and sudden unexpected death in epilepsy (SUDEP). We sought to evaluate autonomic differences, including heart rate (HR), heart rate variability (HRV) and corrected QT-duration (QTc) between two epileptic (EB1, EB2) and one control (CB) baboon, and the autonomic effects of high-frequency (HF) microburst Vagal Nerve Stimulation (VNS) Therapy in the epileptic baboons. At baseline, EB2's HR was increased over both EB1 and CB, and EB1's HRV was decreased compared to the others. QTc-intervals were significantly prolonged in both epileptic baboons. EB1 became free of generalized tonic-clonic seizures (GTCS) with VNS therapy, whereas EB2's GTCS were reduced by a third. HR decreased in both epileptic baboons, but while HRV improved in EB1, it decreased in EB2. EB2 succumbed to SUDEP after 9 months. This pilot study demonstrates abnormalities in HR, HRV and QTc-intervals in epileptic baboons. HF VNS Therapy demonstrated different effects on HRV in the two epileptic baboons, which, in addition to persistent GTCS and elevated HR, may have contributed to SUDEP risk in EB2. Future studies are needed to establish normative values for HRV and determine variability of HR, HRV and QTc-intervals in epileptic baboons.

Research with nonhuman primates (NHPs) – monkeys for the most part – has led to critical health advances that have saved or improved millions of human lives. While NHPs account for just one-half of one percent of animals in current medical research, it is no exaggeration to say they are essential to our ability to find cures for cancer, AIDS, Alzheimer’s, Parkinson’s, obesity/diabetes, and dozens of other diseases that cause human suffering and death.

An increased level of sodium-lithium countertransport (SLC) activity has been associated with salt-sensitive hypertension. Previous findings have suggested that dysregulation of the renin-angiotensin-aldosterone system (RAAS) may be involved in the mechanism linking elevated SLC activity and hypertension. Therefore, baboons with different levels of SLC activity were given two diets differing in sodium content, with and without an angiotensin II (ANG II) infusion, to investigate the relationship between SLC activity, the RAAS, and physiological regulation by sodium. Although we anticipated that high SLC activity would be associated with inappropriate function of the RAAS and greater arterial pressure sensitivity to dietary sodium and ANG II and that low SLC activity would be associated with the least BP sensitivity, we found that the low SLC phenotype correlated with BP sensitivity similar to the high SLC phenotype, and the normal SLC phenotype showed the least BP sensitivity to dietary sodium and ANG II.

RATIONALE:Streptococcus pneumoniae is the leading cause of community-acquired pneumonia and infectious death in adults worldwide. A non-human primate model is needed to study the molecular mechanisms that underlie the development of severe pneumonia, identify diagnostic tools, explore potential therapeutic targets, and test clinical interventions during pneumococcal pneumonia.OBJECTIVE:To develop a non-human primate model of pneumococcal pneumonia.METHODS:Seven adult baboons (Papio cynocephalus) were surgically tethered to a continuous monitoring system that recorded heart rate, temperature, and electrocardiography. Animals were inoculated with 109 colony-forming units of S. pneumoniae using bronchoscopy. Three baboons were rescued with intravenous ampicillin therapy. Pneumonia was diagnosed using lung ultrasonography and ex vivo confirmation by histopathology and immunodetection of pneumococcal capsule. Organ failure, using serum biomarkers and quantification of bacteremia, was assessed daily.RESULTS:Challenged animals developed signs and symptoms of pneumonia 4 days after infection. Infection was characterized by the presence of cough, tachypnea, dyspnea, tachycardia and fever. All animals developed leukocytosis and bacteremia 24 hours after infection. A severe inflammatory reaction was detected by elevation of serum cytokines, including Interleukin (IL)1Ra, IL-6, and IL-8, after infection. Lung ultrasonography precisely detected the lobes with pneumonia that were later confirmed by pathological analysis. Lung pathology positively correlated with disease severity. Antimicrobial therapy rapidly reversed symptomology and reduced serum cytokines.CONCLUSIONS:We have developed a novel animal model for severe pneumococcal pneumonia that mimics the clinical presentation, inflammatory response, and infection kinetics seen in humans. This is a novel model to test vaccines and treatments, measure biomarkers to diagnose pneumonia, and predict outcomes.

Glucose-dependent insulinotropic polypeptide (GIP) has important actions on whole body metabolic function. GIP and its receptor are also present in the central nervous system and have been linked to neurotrophic actions. Metabolic effects of central nervous system GIP signaling have not been reported. We investigated whether centrally administered GIP could increase peripheral plasma GIP concentrations and influence the metabolic response to a mixed macronutrient meal in nonhuman primates. An infusion and sampling system was developed to enable continuous intracerebroventricular (ICV) infusions with serial venous sampling in conscious nonhuman primates. Male baboons (Papio sp.) that were healthy and had normal body weights (28.9 ± 2.1 kg) were studied (n = 3). Animals were randomized to receive continuous ICV infusions of GIP (20 pmol·kg-1·h-1) or vehicle before and over the course of a 300-min mixed meal test (15 kcal/kg, 1.5g glucose/kg) on two occasions. A significant increase in plasma GIP concentration was observed under ICV GIP infusion (66.5 ± 8.0 vs. 680.6 ± 412.8 pg/ml, P = 0.04) before administration of the mixed meal. Increases in postprandial, but not fasted, insulin (P = 0.01) and pancreatic polypeptide (P = 0.04) were also observed under ICV GIP. Effects of ICV GIP on fasted or postprandial glucagon, glucose, triglyceride, and free fatty acids were not observed. Our data demonstrate that central GIP signaling can promote increased plasma GIP concentrations independent of nutrient stimulation and increase insulin and pancreatic polypeptide responses to a mixed meal.

Measurements of glomerular filtration rate (GFR), and the fractional excretion of sodium (Na) and potassium (K) are critical in assessing renal function in health and disease. GFR is measured as the steady state renal clearance of inulin which is filtered at the glomerulus, but not secreted or reabsorbed along the nephron. The fractional excretion of Na and K can be determined from the concentration of Na and K in plasma and urine. The renal clearance of inulin can be demonstrated in an anesthetized animal which has catheters in the femoral artery, femoral vein and bladder. The equipment and supplies used for this procedure are those commonly available in a research core facility, and thus makes this procedure a practical means for measuring renal function. The purpose of this video is to demonstrate the procedures required to perform a lab demonstration in which renal function is assessed before and after a diuretic drug. The presented technique can be utilized to assess renal function in rat models of renal disease.

SESSION TITLE: Critical Care Posters I SESSION TYPE: Original Investigation Poster PRESENTED ON: Wednesday, October 28, 2015 at 01:30 PM - 02:30 PM PURPOSE: Cardiac complications during pneumococcal pneumonia could explain up to 40% of deaths related with pneumonia. Our research group have described recently that Streptococcus pneumoniae is able to translocate into the heart of experimentally infected mice and disrupt its normal function during invasive pneumococcal disease. To translate these findings to humans there is a need to identify non-invasive methods that could assess cardiac injury during and post-infection. Cardiac troponins are highly sensitive and specific markers of myocardial injury. Our aim was to test the kinetics of troponin I levels in an experimental non-human primate model of pneumococcal pneumonia. METHODS: Healthy young baboons (n=3, 12-13 years old) were assessed at baseline with transthoracic 2D echocardiogram and 12-lead electrocardiogram evaluation. Baboons were tethered to allow for continuous electrocardiogram (ECG), heart rate, blood sampling, and temperature monitoring during infection. Anesthetized baboons were bronchoscopically infected with S. pneumoniae (108 CFU). Myocardial injury was tested by daily ultrasensitive troponin I ELISA. Baboons were euthanized when they developed a moribund state or 10 days post-infection. Pneumococcal myocardial invasion was assessed by fluorescent microscopy. RESULTS: Baboons developed mild, moderate and severe pneumonia. The baboon with mild pneumonia had transient bacteremia and only one foci of bronchopneumonia. Baboons with moderate and severe pneumonia progressed to have extensive lobar and multilobar pneumonia, respectively. The baboon with severe pneumonia died four days post-infection. Pneumococcal invasion to the myocardium was observed in the baboons with moderate and severe disease. Among the three baboons there was a progressive incremental troponin I levels from baseline to day 4 post-infection. The highest level of troponin was observed among the baboons with moderate and severe pneumonia (0.066 vs. 0.015; p=0.03). Continues ECG revealed sinus tachycardia and non-specific changes of myocardial injury. CONCLUSIONS: Troponin elevation increased in baboons with pneumonia in the presence of S. pneumoniae myocardial translocation. CLINICAL IMPLICATIONS: The mechanism of myocardial invasion and cardiac complications during pneumococcal pneumonia needs to be elucidated to identify possible therapeutic targets. DISCLOSURE: The following authors have nothing to disclose: Marcos Restrepo, Luis Reyes, Cecilia Hinojosa, Jessica Perry, Robert Shade, Nilam Soni, Melissa De La Garza, Luis Giavedoni, Carlos Orihuela No Product/Research Disclosure Information

Abstract Introduction Prostate cancer (PCa) is the second most common cancer in American men, resulting in 32,000 deaths annually. Challenges for the management of PCa include the inability to distinguish indolent from aggressive and untreatable castration-resistant PCa. These challenges can be addressed by understanding the molecular basis of cancer initiation and progression. Prostate specific antigen (PSA) is widely used for screening, diagnosing and monitoring of PCa, although it is also elevated in other disorders and therefore lacks specificity and sensitivity. Alternative splicing is a common mechanism used in nature to enhance protein diversity. Alternative splicing of the PSA gene produces at least 15 transcripts of 0.7-6.1 kb (Heuzé-Vourc'h et al 2003). Several variants of PSA have been reported and are shown to be differentially expressed in PCa compared to normal prostates or to benign prostatic hyperplasia (BPH). Here we describe the identification of four variants of PSA with the potential to improve on the specificity of the PSA test. Material and Methods Human prostate cDNAs were from cryopreserved prostate tissues collected after radical prostatectomies and have been described previously (Mubiru et al. 2004). PCR was carried out using primers that amplify the complete PSA mRNA and also those specific for the most common splice variant (Heuzé-Vourc'h et al. 2001). The PCR products were inserted into the PCR2.1-TOPO vector (Life Technologies) and sequenced. Results We identified four novel splice variants of the PSA gene. Novel PSA Splice VariantsTypeNameContaining Exons (Ex) and Introns (In)Molecular Weight (Da)pIPSA mRNAPSAEx3del246Ex1, Ex2, Partial Ex38,5949.61PSA mRNAPSA25RVEx1, Partial In111,1815.55PSA mRNAPSA Yang 28Ex1, Ex2, Ex514,1539.59PSA mRNAPSA Yang 21Ex1, Ex2, Partial Ex3, Ex4, Ex523,6986.64 Results also indicate that some transcripts are differentially expressed in tumors compared to non-tumors. References 1.Mubiru JN, et al. (2004). Gene 327: 89-98. 2.Heuzé-Vourc'h N, et al. (2003) Eur J Biochem 270: 706-714. 3.Heuzé-Vourc'h N, et al. (2001) Eur J Biochem 268: 4408-4413. Citation Format: Alice S. Yang, Thushari Alahapperuma, James N. Mubiru, Magdalena Garcia-Forey, Robert E. Shade. PSA splice variants and their expression in prostate cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2347. doi:10.1158/1538-7445.AM2014-2347

Abstract Background: Adult T-cell leukemia/lymphoma (ATLL) is a T-cell lymphoma that can be found in the blood (leukemia), lymph nodes (lymphoma), or many areas of the body. ATLL has been linked to infection by the human T-cell lymphotropic virus type 1 (HTLV-1). HTLV-1 infects millions of people worldwide. However, less than five percent of individuals with the HTLV-1 will develop HTLV-associated diseases such as ATLL (d'Offay et al. 2007). The simian counterpart of HTLV-1 is the Simian T-cell leukemia virus-1 (STLV-1). STLV-1 is endemic in Old World monkeys including baboons, and has been linked to lymphomas in nonhuman primates (Takemura et al 2002, Saksena et al. 1994, d'Offay et al. 2007). Lymphoma is the most common neoplasm of nonhuman primates (Hubbard et al. 1993, Hunt et al 1983). Some baboons naturally infected with STLV-1 develop leukemia/lymphoma that shares clinical and pathological features with human ATLL (Hubbard et al. 1993, Schatzl et al 1993). At Texas Biomed we have a large population of pedigreed baboons, some are infected with STLV-1. The aim of our study was to investigate the relationship between STLV-1 infection and ATLL in a captive baboon colony and to develop the baboon as a model to study the link between retroviruses and ATLL. Methods: We reviewed the pathology reports of 135 baboons that went for necropsy during the years of 1991-2014 that had a diagnosis of lymphosarcoma which involved non-lymphoid organs. Results: The most common clinical finding was generalized lymphadenopathy and weight loss. Microscopic examination indicated that in most animals neoplastic cells were found in most organs. Organs were effaced by a uniform population of small, well differentiated lymphocytes with occasional mitoses. Also, the lymph nodes and the splenic white pulp contained increased numbers of lymphocytes with multifocal disorganization of the preexistent architecture. Other than the lymphoid organs, the lungs were the organ involved most (>90%). Other organs involved included kidney (56%), liver (60%), heart (46%), skin (9%) and eye (3%). Using Immunohistochemistry the neoplastic cells showed strong immunoreactivity for CD3 but were negative for CD79a and CD68. Conclusions: Due to the fact that baboons naturally infected with STLV-1 develop leukemia/lymphoma that shares clinical and pathological features with human ATLL, we recommend using the baboon as a useful model for studying the mode of transmission and therapies for ATLL. Further studies: We have initiated a study on the role of genetics in the pathogenicity STLV-1. References: Hubbard GB, Moné JP et al.1993; 43(4):301-9. d'Offay JM, Eberle R et al. 2007;57(1):105-14. Takemura T, Yamashita M et al. 2002;76(4):1642-8. Saksena NK, Herve V et al.1994;198(1):297-310. Hunt RD, Blake BJ et al.1983; 80(16):5085-9. Schätzl H, Tschikobava M et al.1993;7 Suppl 2:S86-92. Courgnaud V, Van Dooren S et al. 2004;78(9):4700-9. Citation Format: Thushari Alahapperuma, James N. Mubiru, Alice Yang, Maggie Garcia-Forey, Edward J. Dick, Michael Owston, Robert E. Shade. Development of the baboon as an animal model for adult T-cell leukemia/lymphoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 82. doi:10.1158/1538-7445.AM2014-82