Activation of myeloid and endothelial cells by CD40L gene therapy supports T-cell expansion and migration into the tumor microenvironment.

CD40 is an interesting target in cancer immunotherapy due to its ability to stimulate T-helper 1 immunity via maturation of dendritic cells and to drive M2 to M1 macrophage differentiation. Pancreatic cancer has a high M2 content that has shown responsive to anti-CD40 agonist therapy and CD40 may thus be a suitable target for immune activation in these patients. In this study, a novel oncolytic adenovirus armed with a trimerized membrane-bound extracellular CD40L (TMZ-CD40L) was evaluated as a treatment of pancreatic cancer. Further, the CD40L mechanisms of action were elucidated in cancer models. The results demonstrated that the virus transferring TMZ-CD40L had oncolytic capacity in pancreatic cancer cells and could control tumor progression. TMZ-CD40L was a potent stimulator of human myeloid cells and T-cell responses. Further, CD40L-mediated stimulation increased tumor-infiltrating T cells in vivo, which may be due to a direct activation of endothelial cells to upregulate receptors for lymphocyte attachment and transmigration. In conclusion, CD40L-mediated gene therapy is an interesting concept for the treatment of tumors with high levels of M2 macrophages, such as pancreatic cancer, and an oncolytic virus as carrier of CD40L may further boost tumor killing and immune activation.

Monitoring of intrathecal oxygen tension during experimental aortic occlusion predicts ultrastructural changes in the spinal cord.

OBJECTIVE: To study the correlation between intrathecal PO2 and ultrastructural changes in the spinal cord during thoracic aortic occlusion in pigs. MATERIAL AND METHODS: In 18 pigs, online intrathecal oxygenation was monitored by a multiparameter Paratrend catheter (Biomedical Sensors, High Wycombe, United Kingdom) during 60 minutes' clamping of the proximal and distal descending thoracic aorta. The animals were randomly divided into 2 groups (A and B) depending on the level of distal aortic clamping. Distal aortic perfusion was restored through an aorto-iliac shunt, which also maintained low thoracic segmental perfusion of the spinal cord in group B. Perfusion-fixation technique was used before harvesting the spinal cord specimens, which later were evaluated with light and electron microscopy by an independent observer. Intrathecal parameters were interpreted as normal if PO2 was more than 0.8 kPa and PCO2 was less than 12 kPa, as intermediate ischemia if PO2 was 0.8 or less or PCO (2) was more than 12 kPa, and as absolute ischemia if PO2 was 0.8 or less and PCO2 was more than 12 kPa. RESULTS: Among 6 animals with ultrastructural changes of absolute spinal cord ischemia-reperfusion injury, 5 also had absolute ischemia according to variables derived by the Paratrend catheter. The 2 methods were in agreement in 3 of 5 animals with intermediate ischemia-reperfusion changes and in 5 of 6 animals with normal findings. The accuracy of cerebrospinal fluid PO2 and PCO2 to predict electron microscopy-verified intermediate or absolute ischemia-reperfusion injury was 94%. CONCLUSIONS: Monitoring of intrathecal PO2 after clamping of the descending aorta correlated with ultrastructural changes in the spinal cord in this pig model.

Aspects of the spinal cord circulation as assessed by intrathecal oxygen tension monitoring during various arterial interruptions in the pig.

OBJECTIVE: We sought to study the effect of various modes of interruption of the spinal cord blood supply on intrathecal oxygenation. METHODS: In 24 pigs intrathecal PO (2), PCO (2), and pH were continuously monitored with a multiparameter catheter (Paratrend 7, Biomedical Sensors; Diametrics Medical, Inc, St Paul, Minn) during and after aortic crossclamping or selective interruption of segmental arteries and proximal collateral circulation. RESULTS: Proximal aortic clamping (n = 6) produced complete ischemia, whereas a second clamp close to the celiac trunk (n = 4) partly protected against spinal cord ischemia. This is explained by prevention of the steal phenomenon in the excluded part of the aorta. Adding clamps to the subclavian arteries (n = 6) created complete spinal ischemia as the collateral circulation was interrupted. In another group (n = 4) all segmental arteries below T5 were occluded with no reaction in the intrathecal variables. Additional selective clamping of supreme intercostal arteries (n = 4) showed the relative importance of the subclavian and vertebral collateral pathways. CONCLUSIONS: Continuous intrathecal PO (2) was monitored during various modes of interruption of the spinal cord blood supply. This provided insight into the ischemia mechanisms and relative importance of the segmental contribution and proximal collateral pathways of the spinal cord circulation in pigs. A short literature review is given, and aspects of comparative anatomy are discussed.

A new method of intrathecal PO2, PCO2, and pH measurements for continuous monitoring of spinal cord ischemia during thoracic aortic clamping in pigs.

BACKGROUND: Impaired spinal cord circulation during thoracic aortic clamping may result in paraplegia. Reliable and fast responding methods for intraoperative monitoring are needed to facilitate the evaluation of protective measures and efficiency of revascularization. METHODS: In 11 pigs, a multiparameter PO2, PCO2, and pH sensor (Paratrend 7, Biomedical Sensors Ltd, United Kingdom) was introduced into the intrathecal space for continuous monitoring of cerebrospinal fluid (CSF) oxygenation during thoracic aortic cross-clamping (AXC) distal to the left subclavian artery. A laser-Doppler probe was inserted into the epidural space for simultaneous measurements of spinal cord flux. Registrations were made before and 30 minutes after clamping and 30 and 60 minutes after declamping. The same measuring points were used for systemic hemodynamic and metabolic data acquisition. RESULTS: The mean CSF PO2 readings of 41 mm Hg (5.5 kPa) at baseline decreased within 3 minutes to 5 mm Hg (0.7 kPa) during AXC (P < .01). Spinal cord flux measurement responded immediately in the same way to AXC. Both methods indicated normalization of circulation during declamping. Significant (P < .01) changes were also observed in the CSF metabolic parameters PCO2 and pH. CONCLUSIONS: In this experimental model of spinal ischemia by AXC, online monitoring of intrathecal PO2, PCO2, and pH showed significant changes and correlated well with epidural laser-Doppler flowmetry (P < .01).

Influence of low proximal aortic pressure on spinal cord oxygenation in experimental thoracic aortic occlusion.

BACKGROUND: To evaluate the effect of low proximal aortic pressure on cerebrospinal fluid (CSF) oxygenation in an experimental thoracic occlusion model. METHODS: In nine pigs, continuous intrathecal pO(2), pCO(2) and pH monitoring was used during double descending thoracic aortic clamping following insertion of an aorto-aortic shunt. In five pigs, the shunt was connected to a citrated bag adjusted at approximately 40-45 cm above the heart for partial exsanguination in order to decrease mean proximal aortic pressure (MPAP) to below 50 mmHg. In four animals, sodium nitroprusside infusion was used for this purpose. RESULTS: Intrathecal pO(2) demonstrated a significant decrease from 4.9+/-2.1 to 2.9+/-2.4 kPa after 10 minutes of aortic cross-clamping. Lowering proximal aortic pressure caused a further significant decrease to 1.2+/-1.7 kPa (p<0.05). In seven pigs (5 in the exsanguination and 2 in the vasodilator group), restoration of mean proximal aortic pressure to 94.0+/-27.7 caused a recovery of CSF pO(2) from 1.2+/-1.9 to 2.8+/-3.0 (p<0.05). CONCLUSIONS: The results of this study demonstrate that MPAP which provides spinal cord perfusion through subclavian-vertebral arteries are crucial for maintenance of spinal cord oxygenation during thoracic aortic occlusion in this pig model.

Intraperitoneal tonometry for detection of regional enteric ischaemia.

BACKGROUND: The objective of this study was to test the hypothesis that intraperitoneal tonometry can be a specific monitor for ischaemia in the small intestine. METHODS: Twelve pigs were anaesthetized and mechanically ventilated. The celiac artery (CA), the superior mesenteric artery (SMA) and the inferior mesenteric artery (IMA) were identified. Tonometry catheters were positioned intraperitoneally at three different locations where blood supply varied. One at a time of the mesenteric arteries was occluded, producing regional ischaemia in different splanchnic organs. RESULTS: Regional PCO2 (Pr CO2) increased significantly in the intestinal region, in the small intestine, only during the SMA clamping. In the epigastric region, i.e. in the space between the liver and the stomach, PrCO2 increased significantly only during CA clamping. CONCLUSION: Intraperitoneal tonometry in the intestinal region can be a specific monitor of ischaemia in the small intestine. INVESTIGATION: The care and handling of the animals was in accordance with legislation by the Swedish Board of Agriculture.

Intraperitoneal and sigmoid colon tonometry in porcine hypoperfusion and endotoxin shock models.

BACKGROUND: The objective of the present study was to assess the accuracy of an air tonometry device in vivo within a wide range of regional carbon dioxide tension (PrCO2) values by using saline tonometry as the standard and to investigate the possibilities to monitor perfusion of the intestine by tonometry in the intraperitoneal cavity. METHODS: Piglets were anesthetized and mechanically ventilated. A pair of tonometry catheters was placed in the sigmoid colon, while another pair was placed intraperitoneally in the right lower quadrant of the abdomen. Air tonometric regional PCO2 (aPrCO2) was measured every 15 min intraperitoneally and every 20 min in the sigmoid colon. Saline tonometric measurements were made every 30 min and steady-state values (ssPrCO2) were derived. Hypoperfusion shock was induced by graded constriction of the aorta. Endotoxin shock was induced by administration of lipopolysaccharide (LPS). RESULTS: The obtained average PrCO2 (=matched (aPrCO2+ ssPrCO2)/2) values ranged from 5.1 kPa to 14.7 kPa. Regional air PCO2 (aPrCO2) and steady-state saline PCO2 (ssPrCO2) exhibited a strong positive linear relationship (r=0.959). The 95% confidence interval of the mean of dPrCO2 (=aPrCO2-ssPrCO2) was 0.31-0.46 kPa. Intraperitoneal tonometric PrCO2 was lower than intraluminal PrCO2 in the sigmoid colon, and was also more sensitive to circulatory changes than sigmoid colon PrCO2. CONCLUSION: The regional air PCO2 (aPrCO2) showed good agreement with the steady-state saline regional PCO2 (ssPrCO2). Intraperitoneal measurements may be an alternative method of monitoring intestinal perfusion after abdominal surgery.

Increased FiO2 improves intrathecal oxygenation during thoracic aortic cross-clamping in pigs.

OBJECTIVES: To investigate the effect of 100% oxygen ventilation on cerebrospinal fluid (CSF) oxygenation in 11 pigs during thoracic aortic cross-clamping. DESIGN: An aorto-aortic shunt was used for control of central hemodynamics and study of hypoperfusion by exsanguination. CSF PO2, PCO2 and pH were continuously monitored before and during clamping. The changes in hemodynamic parameters and intrathecal gas tensions in response to variations in proximal mean aortic pressure and fraction of inspired oxygen (FiO2) were recorded. RESULTS: Baseline CSF PO2 decreased from 4.8 +/- 1.9 to 2.6 +/- 2.2 kPa following aortic occlusion. Increasing FiO2 to 1.0 resulted in a significant increase in CSF PO2 to 4.1 +/- 3.0 with a return to 2.7 +/- 2.1 kPa after reducing FiO2 to 0.4 again. The same variations in FiO2 did not induce any significant changes in CSF PO2 during hypotension. CONCLUSION: Increased FiO2 during experimental thoracic aortic cross-clamping with stable proximal arterial pressure helps to maintain CSF PO2, whereas severe hypotension could not be compensated for by hyperoxemia.

Relationship between intrathecal oxygen tension and ultrastructural changes in the spinal cord during experimental aortic clamping.

OBJECTIVES: To investigate spinal cord ultrastructure related to cerebrospinal fluid (CSF) oxygenation. DESIGN: experimental aortic occlusion model with intrathecal oxygen tension monitoring. MATERIALS AND METHODS: Two groups of pigs underwent proximal (P) or double (D) aortic occlusion for 30 min followed by 1 h of reperfusion. In a third group (I) segmental arteries distal to T3 were clamped for 90 min. A thin pO(2), pCO(2) and pH sensor was placed intrathecally for continuous monitoring of CSF. Spinal cord segments were studied by electron microscopy (EM). RESULTS: In group P, CSF-pO(2)rapidly decreased during clamping and major changes in pH and pCO(2)were seen. EM demonstrated neuronal degeneration with loss of cellular integrity and severe affection of organelles. In the group D, CSF oxygenation decreased to about half, but with only moderate changes in the metabolic parameters. Group I showed no significant changes in CSF measurements. The latter groups were similar at EM, showing only mild mitochondrial changes. CONCLUSIONS: The level of CSF oxygenation during aortic cross-clamping or segmental artery interruption seems to correlate with ultrastructural changes in the spinal cord. This online intrathecal monitoring technique may provide valuable information on spinal cord circulation during thoracoabdominal aortic surgery.

Influence of segmental spinal cord perfusion on intrathecal oxygen tension during experimental thoracic aortic crossclamping.

PURPOSE: The purpose of this study was to evaluate the possibility of identifying alterations in blood supply to the spinal cord during thoracic aortic crossclamping. METHODS: In 17 pigs, a multiparameter PO(2), PCO(2,) and pH sensor was introduced into the intrathecal space for continuous monitoring of cerebrospinal fluid (CSF) oxygenation during aortic crossclamping. An epidural laser Doppler probe was used to measure spinal cord flux. After insertion of an aortic shunt from the left subclavian to the left iliac artery and interruption of the right subclavian and lumbar arteries (L2-L5), the thoracic aorta just distal to the left subclavian artery was clamped for 60 minutes. By placement of the distal aortic crossclamping below the level of L1 in group A (n = 9 animals), perfusion of only the abdominal visceral arteries was maintained. In group B (n = 8 animals), the distal aortic crossclamping was above the level of T12, and thus some spinal cord perfusion was maintained through the aortic shunt. RESULTS: The significant decrease in CSF PO(2) was observed within 3 minutes after the placement of the proximal aortic crossclamping and was normalized in all animals after establishment of the shunt flow. In group A, distal aortic crossclamping caused a decrease in CSF PO(2) with at least 50% of the preclamping values within 3 minutes. The mean CSF PO(2) of 2.99 +/- 0.70 kPa at 60 minutes of distal aortic crossclamping in group B was significantly higher than in group A (0.11 +/- 0.11 kPa; P <. 001). In group A, PCO(2) measurements showed no significant changes in 3 minutes after distal aortic crossclamping but revealed significantly higher values at 30 and 60 minutes compared with group B. Spinal cord flux values showed similar changes as CSF PO(2) during the whole experiment in both groups. CONCLUSION: In this experimental model of aortic crossclamping, continuous CSF oxygen tension monitoring allows rapid detection of alterations in spinal cord circulation.

A prolonged spinal cord ischaemia model in pigs. Passive shunting offers stable central haemodynamics during aortic occlusion.

OBJECTIVES: to evaluate the effect of a modified aortic shunt on central haemodynamic variables during experimental thoracic aortic occlusion in a prolonged spinal cord ischaemia model. MATERIAL AND METHODS: central haemodynamic variables were evaluated during aortic cross-clamping. In the shunt group (n=11), after the placement of proximal and distal aortic clamps, distal aortic perfusion was restored through an aortoiliac shunt via the left subclavian artery. In the no-shunt group (n=11), spinal cord ischaemia was achieved with only proximal aortic cross-clamping. The clamping time was 60 minutes in the shunt group and 30 minutes in the no-shunt group. RESULTS: in the no-shunt group, all animals needed inotropic support, vasodilators and buffers during the experiment. None of these drugs were needed in the shunt group. In the no-shunt group, cross-clamping caused a significant increase in mean arterial pressure and heart rate compared to baseline values. These variables were stable in the shunt group during aortic occlusion. In the reperfusion period cardiac output, heart rate and arterial pCO(2)were significantly higher in the no-shunt than in the shunt group. CONCLUSION: the present experimental spinal cord ischaemia model, using double aortic cross-clamping with shunt, offers improved central haemodynamics. This enables the study of prolonged selective spinal cord ischaemia without interaction from vasoactive drugs or systemic reperfusion.

Treatment of severe ethylene glycol intoxication with continuous arteriovenous hemofiltration dialysis.

Ethylene glycol is the major component of many antifreeze solutions and its metabolites may cause severe intoxication. Treatment involves correction of metabolic acidosis, ethanol administration and enhancement of elimination. The most commonly used elimination technique is hemodialysis but peritoneal dialysis has also been described. We report a case of severe ethylene glycol poisoning with multiple organ dysfunction. Treatment with continuous arteriovenous hemofiltration dialysis was associated with a favorable outcome and might be an alternative to hemodialysis in similar cases of circulatory instability or in situations where hemodialysis is unavailable.