Traumatic brain injury (TBI) is often accompanied by hemorrhage, and treatment of hemorrhagic shock (HS) after TBI is particularly challenging because the two therapeutic treatment strategies for TBI and HS often conflict. Ischemia/reperfusion injury from HS resuscitation can be exaggerated by TBI-induced loss of autoregulation. In HS resuscitation, the goal is to restore lost blood volume, while in the treatment of TBI the priority is focused on maintenance of adequate cerebral perfusion pressure and avoidance of secondary bleeding. In this study, we investigate the responses to resuscitation from severe HS after TBI in rats, using fresh blood, polymerized human hemoglobin (PolyhHb), and lactated Ringer’s (LR). Rats were subjected to TBI by pneumatic controlled cortical impact. Shortly after TBI, HS was induced by blood withdrawal to reduce mean arterial pressure (MAP) to 35–40 mmHg for 90 min before resuscitation. Resuscitation fluids were delivered to restore MAP to ~ 65 mmHg and animals were monitored for 120 min. Increased systolic blood pressure variability (SBPV) confirmed TBI-induced loss of autoregulation. MAP after resuscitation was significantly higher in the blood and PolyhHb groups compared to the LR group. Furthermore, blood and PolyhHb restored diastolic pressure, while this remained depressed for the LR group, indicating a loss of vascular tone. Lactate increased in all groups during HS, and only returned to baseline level in the blood reperfused group. The PolyhHb group possessed lower SBPV compared to LR and blood groups. Finally, sympathetic nervous system (SNS) modulation was higher for the LR group and lower for the PolyhHb group compared to the blood group after reperfusion. In conclusion, our results suggest that PolyhHb could be an alternative to blood for resuscitation from HS after TBI when blood is not available, assuming additional testing demonstrate similar favorable results. PolyhHb restored hemodynamics and oxygen delivery, without the logistical constraints of refrigerated blood.

Background: Mechanical overload is an established contributor to the remodeling of T-tubules (TT) and associated dyssynchronous Ca 2+ release leading to abnormal excitation-contraction coupling in cardiomyopathies. We examined the induction of pathologic TT remodeling in isolated cardiomyocytes and the involvement of microtubules in this process. Methods: Normal adult rat ventricular myocytes (aRVMs) were cultured for 48hrs on MREs at either physiological (10kPa) or pathological (50kPa) stiffness. Variable in vitro load application was achieved using custom magnetorheologic elastomers (MREs) with tunable stiffness. Cells were stained with Di-8-ANEPPS and Fluo-4-AM and imaged using Airyscan confocal microscopy. Microtubule dependency was investigated by treating cells with 1μM nocodazole, a microtubule depolymerization agent.Structural changes in TT networks were quantified in terms of regularity, luminal dimensions, density, and directionality. Field stimulation of cells allowed for acquisition of [Ca 2+ ] i transients via line scans and subsequent analysis of key [Ca 2+ ] i transient parameters. Changes in protein expression were determined via Western blotting. Results: Compared to the physiological stiffness group, aRVMs subjected to pathologically increased stiffness exhibited decreased density and regularity of the TT system, and increased luminal dimension. Direction-specific density analysis showed that pathologically loaded cells have more longitudinal tubule elements and fewer transverse tubule elements. This remodeling was attenuated in aRVMs treated with nocodazole in terms of regularity, luminal diameter, and direction-specific density. The observed changes in both Ca 2+ handling parameters and key protein expression resembled those typical of the in vivo myopathic phenotype. Conclusion: The results suggest that load-induced TT remodeling occurs via a cardiomyocyte autonomous, microtubule-dependent mechanism.

Introduction Heart failure is associated with degradation of cell functions and extracellular matrix proteins, but the trigger mechanisms are uncertain. Our recent evidence shows that active digestive enzymes can leak out of the small intestine into the systemic circulation and cause cell dysfunctions and organ failure. Methods Accordingly, we investigated in morning fasting plasma of heart failure (HF) patients the presence of pancreatic trypsin, a major enzyme responsible for digestion. Results Western analysis shows that trypsin in plasma is significantly elevated in HF compared to matched controls and their concentrations correlate with the cardiac dysfunction biomarker BNP and inflammatory biomarkers CRP and TNF-α. The plasma trypsin levels in HF are accompanied by elevated pancreatic lipase concentrations. The trypsin has a significantly elevated activity as determined by substrate cleavage. Mass spectrometry shows that the number of plasma proteins in the HF patients is similar to controls while the number of peptides was increased about 20% in HF patients. The peptides are derived from extracellular and intracellular protein sources and exhibit cleavage sites by trypsin as well as other degrading proteases (data are available via ProteomeXchange with identifier PXD026332). Connclusions These results provide the first evidence that active digestive enzymes leak into the systemic circulation and may participate in myocardial cell dysfunctions and tissue destruction in HF patients. Conclusions These results provide the first evidence that active digestive enzymes leak into the systemic circulation and may participate in myocardial cell dysfunctions and tissue destruction in HF patients.

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Hemorrhagic Shock (HS) after Traumatic Brain Injury (TBI) is common and lethal among military and civilians. Blood transfusion has been the resuscitation gold standard procedure for many years. However, only 3% of age‐eligible individuals donate blood, which limits blood availability and cannot meet current blood demands. This study was compared the efficacy to resuscitate from HS after TBI via infusion of Lactated Ringer’s solution (LR, electrolyte solution used to restore the loss of blood volume), fresh blood (Blood, autologous blood drawn during the hemorrhage), or Polymerized Hemoglobin [PolyHb, a Hemoglobin Based Oxygen Carrier (HBOC) that shows promise in increasing oxygenation]. Fifteen Wistar rats (350–400g) were instrumented with catheters in the femoral artery and vein and subjected to TBI through a pneumatic controlled cortical impact (CCI) (Leica Biosystems, Vista, CA). To induce TBI, a 5mm craniotomy over the right cerebral cortex was performed, and the scalp was impacted at a velocity of 5m/s and dwell time of 200ms. The scalp was subsequently closed and animals were given 10 min to stabilize before performing HS. HS was induced by blood withdrawal from the femoral artery catheter to achieve a mean arterial pressure (MAP) between 35–40mmHg, and the hypotensive state was maintained for 90 min before resuscitation. Animals were separated into 3 groups based on the resuscitation solution: Blood, LR, or PolyHb. Animals were monitored for an additional 120 min before euthanasia. Measurements were taken at baseline (BL), 90 min into HS (HS), and 30 min (R1) and 2 hours (R2) after resuscitation. The MAP, systolic blood pressure (SBP), and diastolic blood pressure (DBP) were decreased equally in all groups during HS. Blood and PolyHb groups increased MAP compared to the LR group at R1. The MAP was not different between Blood and PolyHb at R2. The SBP was increased only for the Blood group as compared to the LR group at R1. Moreover, DBP was increased for both Blood and PolyHb compared to the LR group at R1 and R2. As expected, hematocrit decreased in all groups after HS, with only the Blood group recovering hematocrit after resuscitation. Total hemoglobin (tHb) decreased after HS, and increased after resuscitation for the Blood and PolyHb groups but not in the LR group. Oxygen saturation was decreased for the Blood and PolyHb groups compared to the LR group at R1 and R2. The oxygen saturation was increased on the Blood group compared to PolyHb group at R2. After HS all groups increased lactate, after R1 just Blood and LR group recovered lactate to BL levels, and PolyHb did not decreased lactate concentration at R2 compared to the blood group. In conclusion, our results show that PolyHb was effective to resuscitation from HS after TBI, restoring blood pressure similarly to Blood group and could be a good alternative to transfusion for field resuscitation from HS after TBI. Support or Funding Information This work was supported by the NIH Heart Lung and Blood Institute under Grant, R01‐HL126945, and the DOD ...