Hyperbaric oxygen therapy induces kidney protection in an ischemia/reperfusion model in rats.

Ischemia/reperfusion (I/R) injury remains a major cause of graft dysfunction, which impacts short- and long-term follow-up. Hyperbaric oxygen therapy (HBO), through plasma oxygen transport, has been currently used as an alternative treatment for ischemic tissues. The aim of this study was to analyze the effects of HBO on kidney I/R injury model in rats, in reducing the harmful effect of I/R. The renal I/R model was obtained by occluding bilateral renal pedicles with nontraumatic vascular clamps for 45 minutes, followed by 48 hours of reperfusion. HBO therapy was delivered an hypebaric chamber (2.5 atmospheres absolute). Animals underwent two sessions of 60 minutes each at 6 hours and 20 hours after initiation of reperfusion. Male Wistar rats (n = 38) were randomized into four groups: sham, sham operated rats; Sham+HBO, sham operated rats exposed to HBO; I/R, animals submitted to I/R; and I/R+HBO, I/R rats exposed to HBO. Blood, urine, and kidney tissue were collected for biochemical, histologic, and immunohistochemical analyses. The histopathological evaluation of the ischemic injury used a grading scale of 0 to 4. HBO attenuated renal dysfunction after ischemia characterized by a significant decrease in blood urea nitrogen (BUN), serum creatinine, and proteinuria in the I/R+HBO group compared with I/R alone. In parallel, tubular function was improved resulting in significantly lower fractional excretions of sodium and potassium. Kidney sections from the I/R plus HBO group showed significantly lower acute kidney injury scores compared with the I/R group. HBO treatment significantly diminished proliferative activity in I/R (P < .05). There was no significant difference in macrophage infiltration or hemoxygenase-1 expression. In conclusion, HBO attenuated renal dysfunction in a kidney I/R injury model with a decrease in BUN, serum creatinine, proteinuria, and fractional excretion of sodium and potassium, associated with reduced histological damage.

Exploring metabolic pathways in genome-scale networks via generating flux modes.

MOTIVATION: The reconstruction of metabolic networks at the genome scale has allowed the analysis of metabolic pathways at an unprecedented level of complexity. Elementary flux modes (EFMs) are an appropriate concept for such analysis. However, their number grows in a combinatorial fashion as the size of the metabolic network increases, which renders the application of EFMs approach to large metabolic networks difficult. Novel methods are expected to deal with such complexity. RESULTS: In this article, we present a novel optimization-based method for determining a minimal generating set of EFMs, i.e. a convex basis. We show that a subset of elements of this convex basis can be effectively computed even in large metabolic networks. Our method was applied to examine the structure of pathways producing lysine in Escherichia coli. We obtained a more varied and informative set of pathways in comparison with existing methods. In addition, an alternative pathway to produce lysine was identified using a detour via propionyl-CoA, which shows the predictive power of our novel approach. AVAILABILITY: The source code in C++ is available upon request.

Systemic and regional hemodynamic effects of enalaprilat infusion in experimental normotensive sepsis.

Angiotensin-converting enzyme inhibitors have been shown to improve splanchnic perfusion in distinct shock states. We hypothesized that enalaprilat potentiates the benefits of early fluid resuscitation in severe experimental sepsis, particularly in the splanchnic region. Anesthetized and mechanically ventilated mongrel dogs received an intravenous infusion of live Escherichia coli over a period of 30 min. Thereafter, two interventions were performed: fluid infusion (normal saline, 32 mL/kg over 30 min) and enalaprilat infusion (0.02 mg kg(-1) min(-1) for 60 min) in randomized groups. The following groups were studied: controls (fluid infusion, N = 4), E1 (enalaprilat infusion followed by fluid infusion, N = 5) and E2 (fluid infusion followed by enalaprilat infusion, N = 5). All animals were observed for a 120 min after bacterial infusion. Mean arterial pressure, cardiac output (CO), portal vein blood flow (PVBF), systemic and regional oxygen-derived variables, and lactate levels were measured. Rapid and progressive reductions in CO and PVBF were induced by the infusion of live bacteria, while minor changes were observed in mean arterial pressure. Systemic and regional territories showed a significant increase in oxygen extraction and lactate levels. Widening venous-arterial and portal-arterial pCO2 gradients were also detected. Fluid replacement promoted transient benefits in CO and PVBF. Enalaprilat after fluid resuscitation did not affect systemic or regional hemodynamic variables. We conclude that in this model of normotensive sepsis inhibition of angiotensin-converting enzyme did not interfere with the course of systemic or regional hemodynamic and oxygen-derived variables.

Mechanisms of action of hypertonic saline resuscitation in severe sepsis and septic shock.

Small volumes of 7.5% NaCl (2400mOsm/L) have been extensive evaluated in animal models of hemorrhagic shock and in clinical trials of post-traumatic hypotension and as volume support for complex cardiovascular procedures. Hypertonic solutions promote immediate blood volume expansion, restore cardiac output and regional blood flows, improve microcirculation and modulate immune responses, thereby decreasing inflammatory responses triggered by shock and trauma. A large number of very interesting in vivo and in vitro experiments highlighted that hypertonic saline resuscitation may decrease susceptibility to post-traumatic sepsis, modulate trauma and sepsis-induced immune dysfunction, inflammatory response and apoptosis. All those long-term benefits associated with hypertonic resuscitation may be of potential relevance for the management of severe sepsis and septic shock In this review, we describe the mechanisms of action of hypertonic saline based on experimental studies as well as its efficacy and safety based on its clinical use. We believe those studies support the need for additional experimental and clinical studies before the widespread use of hypertonic solutions for the treatment of severe sepsis and septic shock.

Lymphscintigraphic evaluation of manual lymphatic drainage for lower extremity lymphedema.

To evaluate the effect of manual lymphatic drainage on technetium-99m-labeled dextran (99mTcDx) transport, 16 patients with lymphedema of lower extremities underwent two lymphscintigraphy exams by injecting 99mTcDx intradermally into the first interdigital space of the affected extremity. The first was a control examination at rest followed by an examination which included a manual lymphatic drainage session after the injection of the 99mTcDx. Images were obtained 45 minutes and three hours after the injection of the radioisotope. Extremity volumes were also measured before and after the drainage session. The findings from the examinations were assessed in a quantitative, semiquantitative and qualitative manner and compared without and with drainage. The analyses of the extremities' circumference before and after the drainage by paired t-test revealed a significant decrease. The analyses of the quantitative, semi-quantitative and qualitative evaluations evidenced no significant difference, without or with drainage, within the 45-minute and three-hour periods. Thus, manual lymphatic drainage caused an effective reduction in the circumference of the extremities but did not have a significant effect in the transport of 99mTcDx.

Anatomical study of the gastrocnemius venous network and proposal for a classification of the veins.

OBJECTIVES: To present a detailed description of the gastrocnemius venous network. DESIGN: Anatomical study in cadavers. MATERIAL AND METHODS: Forty lower limbs from 20 adult male cadavers were studied. All gastrocnemius veins were dissected from the gastrocnemius muscle heads proximally toward their drainage site. RESULTS: Eighty heads of 40 gastrocnemius muscles showed 438 gastrocnemius veins. The number of veins per muscle head varied between 2 and 12. There were 221 gastrocnemius trunks distributed as 95 main gastrocnemius trunks, 81 axial and 45 collateral ones. From the 95 main gastrocnemius trunks, 83 (87%) drained into the popliteal vein. Direct observation of the gastrocnemius venous network allowed us to classify the anatomical distribution as four distinct types. CONCLUSIONS: The majority of main gastrocnemius venous trunks drain into the popliteal vein. There is wide variability in the number of gastrocnemius veins. We propose a classification of four distinct types of anatomical pattern.

Active spleno-femoral shunt avoids splanchnic congestion during portal triad occlusion: an experimental study.

UNLABELLED: Portal triad occlusion (PTO) is often performed during hepatic resections for trauma or malignancies to minimize intraoperative blood loss. The pringle maneuver is also regularly required during liver transplantation. This maneuver leads to temporary hepatic ischemia and may be associated with splanchnic blood flow congestion, promoting undesirable hemodynamic disturbances in some patients. Veno-venous bypass is a useful, easily performed technique that may avoid those deleterious hemodynamic effects of PTO. We tested the hypothesis that an active spleno-femoral shunt maintains hemodynamic stability and promotes complete decompression of the mesenteric bed, avoiding intestinal mucosal blood congestion, during PTO. METHODS: Seven dogs (17.2 +/- 0.9 kg) were subjected to 45 minutes of hepatic ischemia during which there was an active spleno-femoral shunt. Systemic hemodynamics were evaluated through Swan-Ganz and arterial catheters. Splanchnic perfusion was assessed by portal vein blood flow and hepatic artery blood flow (PVBF and HABF, ultrasonic flowprobe), intestinal mucosal-arterial pCO(2) gradient (D(t-a)pCO(2), tonometry), and regional O(2)-derived variables. RESULTS: No significant changes in systemic and regional parameters were observed during the ischemia period. During reperfusion, a significant decrease in mean arterial pressure, PVBF, and arterial pH was observed. A significant increase in ALT and D(t-a)pCO(2) (4.8 +/- 2.5 to 18.9 +/- 3 mm Hg) was also observed following hepatic blood flow restoration. CONCLUSION: Spleno-femoral shunt maintains systemic hemodynamic stability, with an effective decompression of the splanchnic bed during portal triad occlusion. The deleterious hemodynamic and metabolic effects observed during reperfusion period, such as transitory hypotension, high D(t-a)pCO(2), and acidemia, were associated with an isolated hepatic ischemia-reperfusion injury, not with the blood congestion in the splanchnic bed.

Short-lasting systemic and regional benefits of early crystalloid infusion after intravenous inoculation of dogs with live Escherichia coli.

We investigated the systemic and regional hemodynamic effects of early crystalloid infusion in an experimental model of septic shock induced by intravenous inoculation with live Escherichia coli. Anesthetized dogs received an intravenous infusion of 1.2 x 10(10) cfu/kg live E. coli in 30 min. After 30 min of observation, they were randomized to controls (no fluids; N = 7), or fluid resuscitation with lactated Ringer's solution, 16 ml/kg (N = 7) or 32 ml/kg (N = 7) over 30 min and followed for 120 min. Cardiac index, portal blood flow, mean arterial pressure, systemic and regional oxygen-derived variables, blood lactate, and gastric PCO2 were assessed. Rapid and progressive cardiovascular deterioration with reduction in cardiac output, mean arterial pressure and portal blood flow (approximately 50, approximately 25 and approximately 70%, respectively) was induced by the live bacteria challenge. Systemic and regional territories showed significant increases in oxygen extraction and in lactate levels. Significant increases in venous-arterial (approximately 9.6 mmHg), portal-arterial (approximately 12.1 mmHg) and gastric mucosal-arterial (approximately 18.4 mmHg) PCO2 gradients were also observed. Early fluid replacement, especially with 32 ml/kg volumes of crystalloids, promoted only partial and transient benefits such as increases of approximately 76% in cardiac index, of approximately 50% in portal vein blood flow and decreases in venous-arterial, portal-arterial, gastric mucosal-arterial PCO2 gradients (7.2 +/- 1.0, 7.2 +/- 1.3 and 9.7 +/- 2.5 mmHg, respectively). The fluid infusion promoted only modest and transient benefits, unable to restore the systemic and regional perfusional and metabolic changes in this hypodynamic septic shock model.

Short-lasting systemic and regional benefits of early crystalloid infusion after intravenous inoculation of dogs with live Escherichia coli

We investigated the systemic and regional hemodynamic effects of early crystalloid infusion in an experimental model of septic shock induced by intravenous inoculation with live Escherichia coli. Anesthetized dogs received an intravenous infusion of 1.2 x 10(10) cfu/kg live E. coli in 30 min. After 30 min of observation, they were randomized to controls (no fluids; N = 7), or fluid resuscitation with lactated Ringer's solution, 16 ml/kg (N = 7) or 32 ml/kg (N = 7) over 30 min and followed for 120 min. Cardiac index, portal blood flow, mean arterial pressure, systemic and regional oxygen-derived variables, blood lactate, and gastric PCO2 were assessed. Rapid and progressive cardiovascular deterioration with reduction in cardiac output, mean arterial pressure and portal blood flow (about 50, about 25 and about 70 percent, respectively) was induced by the live bacteria challenge. Systemic and regional territories showed significant increases in oxygen extraction and in lactate levels. Significant increases in venous-arterial (about 9.6 mmHg), portal-arterial (about 12.1 mmHg) and gastric mucosal-arterial (about 18.4 mmHg) PCO2 gradients were also observed. Early fluid replacement, especially with 32 ml/kg volumes of crystalloids, promoted only partial and transient benefits such as increases of about 76 percent in cardiac index, of about 50 percent in portal vein blood flow and decreases in venous-arterial, portal-arterial, gastric mucosal-arterial PCO2 gradients (7.2 ± 1.0, 7.2 ± 1.3 and 9.7 ± 2.5 mmHg, respectively). The fluid infusion promoted only modest and transient benefits, unable to restore the systemic and regional perfusional and metabolic changes in this hypodynamic septic shock model.

Hepatic arterial buffer response fails to restore hepatic oxygenation after temporary liver dearterialization in canines.

BACKGROUND: Hepatic artery thrombosis is a rare but extremely troublesome condition after liver transplantation. Recently, urgent arterial revascularization has been used as rescue therapy, leading to improved graft and patient survivals. Hepatic artery ligation produces a progressive reduction in portal vein blood flow. Theoretically, a hyperemic response may be expected following hepatic artery reperfusion (hepatic artery buffer response, HABR). In this study, we tested the hypothesis that HABR can maintain adequate liver oxygenation after temporary liver dearterialization. METHODS: Seven dogs (19.7 +/- 1.2 kg) subjected to 60 minutes of hepatic artery occlusion were observed for 120 minutes thereafter. Systemic hemodynamics was evaluated through Swan-Ganz and arterial catheters, and splanchnic perfusion by portal vein and hepatic artery blood flows (PVBF and HABF) via an ultrasonic flowprobe. Liver enzymes (ALT and LDH) and systemic and hepatic oxygen delivery (DO2hepat) were calculated using standard formulae. RESULTS: Hepatic artery occlusion induced a progressive reduction in PVBF and DO2hepat. A complete restoration of HABF after hepatic artery declamping was observed; however, the DO2hepat (33.3 +/- 5.9 to 16.5 +/- 5.9 mL/min) did not return to the baseline levels. CONCLUSION: Temporary hepatic artery occlusion induced a progressive decrease in portal vein blood flow during ischemia, an effect that continued during the reperfusion period. The hepatic artery blood flow was promptly restored after declamping. However, HABR was not able to restore hepatic oxygen delivery to baseline levels during the reperfusion period.

Effects of intra-aortic balloon occlusion on intestinal perfusion, oxygen metabolism and gastric mucosal PCO2 during experimental hemorrhagic shock.

BACKGROUND: Aortic occlusion has been suggested for the initial treatment of severe uncontrolled hemorrhagic shock. Our objective is to determine the impact of aortic occlusion, during hemorrhagic shock, on splanchnic mucosal perfusion and to correlate these findings with other systemic and regional markers of splanchnic ischemia. METHODS: Fourteen dogs (17 +/- 1.7 kg) anesthetized with pentobarbital were bled to a mean arterial pressure (MAP) of 40 mm Hg. After 30 min, the animals were randomly assigned to controls (no aortic occlusion, n = 7) and transfemoral aortic occlusion (TAO) at T9 level (n = 7). Superior mesenteric artery blood flow (SMABF, ultrasonic flow probe), gastric mucosal PCO2 (gastric tonometry) and splanchnic oxygen extraction ratio (O2ERsplanc) were evaluated for 120 min. RESULTS: Hemorrhage caused a marked reduction in SMABF and increases in PCO2-gap and O2ERsplanc in both groups. TAO significantly improved MAP and further increased the PCO2-gap and O2ERsplanc, with a decreased SMABF. After reperfusion, SMABF, MAP and O2ERsplanc returned to pre-occlusion values, although the PCO2-gap remained higher in the TAO group. CONCLUSION: Aortic occlusion promotes blood pressure restoration with an additional insult to mucosal perfusion, which could be adequately predicted by global and/or splanchnic oxygen-derived variables during ischemia, but not during the early reperfusion period.

Effects of N-acetylcysteine in hepatic ischemia-reperfusion injury during hemorrhagic shock.

This article seeks to standardize an experimental model of liver ischemia-reperfusion in rats following hemorrhagic shock modulated by N-acetylcysteine (NAC). Twenty-seven adult Wistar rats were randomized into three groups: the HS-IR-Garm underwent hemorrhagic shock with selective hepatic ischemia followed by reperfusion; the HSIR + NAC-G, the same procedure plus NAC; and the control group, only venous catheterization. Blood was withdrawn for 10 minutes until MABP reached 35 mm Hg, which was maintained for 1 hour. The blood was then reinjected as required to maintain MABP at that level. Ringer's lactate solution was infused in a volume equivalent to three times the shed blood, over a period of 15 minutes. Half of the shed blood was reinfused over 5 minutes. HSIR + NAC-G received 150 mg/kg of NAC, during treatment of the shock, and again 10 minutes before reperfusion and continued for 30 minutes. Finally, both groups were subjected to 40 minutes of warm selective hepatic ischemia and reperfusion for 1 hour. Data were analyzed by nonparametric tests (P < or =.05). Liver enzyme levels were higher in HS-IR-G (DHL = 6094 +/- 1688, AST = 746 +/- 175, and ALT = 457 +/- 90) than in HSIR + NAC-G group (DHL = 2920 +/- 284, AST = 419 +/- 113, and ALT = 253 +/- 26). The values in the control group were lower than both experimental groups (DHL = 965 +/- 173, AST = 163 +/- 42, and ALT = 82 +/- 28). Our data showed that liver ischemia-reperfusion injury following hemorrhagic shock produces important hepatic damage and that NAC reduces injury in this rat model.

A prospective comparison of Doppler echocardiography and postoperative angiography in the assessment of left internal thoracic artery graft in 72 patients submitted to minimally invasive direct coronary artery bypass.

BACKGROUND: Concerns have been raised regarding the accuracy of the left internal thoracic artery (LITA) anastomosis performed during minimally invasive direct coronary artery bypass (MIDCAB). In a prospective study, we tested the hypothesis that transthoracic Doppler echocardiography is an adequate technique to determine LITA patency when compared to "gold standard" postoperative angiography. METHODS: Seventy-two consecutive patients with single left anterior descending (LAD) coronary artery stenosis were submitted to MIDCAB performed on a beating heart using the LITA. All patients underwent transthoracic Doppler Echocardiography and angiography before discharge. LITA was considered patent when diastolic fraction (DF) of time-velocity integral was equal or greater than 0.5. FitzGibbon grading system was used to evaluate LITA patency by angiography (A=excellent; B=stenosis reducing caliber of anastomosis or trunk to < 50% of grafted coronary artery; O=occlusion). RESULTS: Angiography showed that LITA was patent in 70 (97.2%) patients, 69 of them been graded A. Adequate image and flow signal of the LITA was achieved in 65 (90.3%) patients, been considered patent in 61 (93.8%) of them. Comparison between echocardiography and angiography in these 65 patients showed a specificity of 96.8% and a sensitivity of 50%. In 7 (9.7%) patients in whom no adequate echocardiography signal was obtained, the LITA graft was normal in six and occluded in one. CONCLUSIONS: For patients whose LITA graft can be imaged, transthoracic Doppler echocardiography is highly specific and is a valuable method for noninvasive evaluation of LITA graft patency after MIDCAB.

Increases in fines and driver licence withdrawal have effectively reduced immediate deaths from trauma on Brazilian roads: first-year report on the new traffic code.

Road accidents are a major cause of death in Brazil, with rates increasing steadily for years. Our objective here is to report the impact of the new Brazilian Traffic Code, introduced in 1998. Its main new features include a large increase in fines and a rigid penalty scoring system that leads to driver license withdrawal. Speed limits have actually been raised on many roads, but adherence to the rules has been monitored more closely. We compare the incidence of injured patients and immediate deaths in road accidents and emergency room admissions to a level I trauma centre in downtown São Paulo between January and December 1998 with corresponding data from between January and December 1997. There was an overall 21.3% reduction in the number of accidents and a 24.7% reduction in immediate deaths, saving 5962 lives on Brazilian highways. Tickets issued fell by 49.5% (601977 during 1997 to 304785 during 1998). Motor vehicle accident-related emergency room admissions decreased by 33.2%. We conclude that very costly tickets and threatened driver licences have proved very effective in decreasing immediate deaths from trauma. Further advances in educational programmes associated with road and vehicle safety measures are likely to provide the much needed further reduction in the still high trauma mortality on Brazilian roads and streets.

Radioisotope blood volume measurement in uncontrolled retroperitoneal haemorrhage induced by a transfemoral iliac artery puncture.

Standard-of-care, large volume crystalloid infusion, in the setting of uncontrolled bleeding, has been challenged and it is not known if fluid resuscitation increases retroperitoneal hemorrhage. We developed an experimental model of retroperitoneal haemorrhage to correlate haemodynamic and metabolic alterations with the blood volume loss. Anaesthetised, spontaneously breathing dogs (17.1+/-0.56 kg) were randomised to unilateral (UL, n=11) or bilateral (BL, n=11) iliac artery puncture, using a metallic device introduced through the femoral arteries and followed for 120 min. Initial and final blood volumes were determined using radioactive tracers, 99mTC and 51Cr, respectively. UL was associated with a stable arterial pressure and a moderate decrease in cardiac output and oxygen delivery. BL induced an abrupt and sustained decrease in mean arterial pressure, from 131.9+/-5.9 to 88.6+/-10.8 mmHg, and a much greater reduction in cardiac output, oxygen delivery and consumption than UL throughout the experiment. Total retroperitoneal blood loss after BL was 36.8+/-3.2 ml/kg, while after UL was 25.1+/-3.4 ml/kg (P=0.0262). We conclude that a transfemoral bilateral iliac artery puncture produces a clinically relevant model of uncontrolled retroperitoneal haemorrhage, with hypotension and low flow state, while a unilateral iliac artery lesion causes a compensated shock state.

Initial management of severe hemorrhage with an oxygen-carrying hypertonic saline solution.

We tested the hypothesis that the combination of polymerized bovine hemoglobin (PBHg) with hypertonic saline may be beneficial for the initial management of hemorrhagic shock in 22 mongrel dogs (15 +/- 1 kg) bled to a mean arterial pressure (MAP) of 40 mm Hg in 5 min and maintained at this level for 45 min (shed blood volume approximately 50 ml/kg). Animals were treated with a 4 ml/kg bolus over 4 min of one of the following fluids: whole blood, 7.5% NaCl (HS), 13 g/dl of PBHg, or 7.5% NaCl combined with polymerized bovine hemoglobin (HS-PBHg). No additional intervention was performed, and the animals were followed for 60 min after treatment. PBHg and HS-PBHg produced a sustained, significant increase in MAP. Cardiac output was transiently increased only after HS and HS-PBHg. A partial increase in superior mesenteric artery blood flow was observed, particularly after HS-PBHg. We concluded that small volumes of PBHg alone restore MAP, but not blood flow. The combination of PBHg with hypertonic saline provides improvements in cardiac output and mesenteric blood flow, suggesting a potential benefit for the initial management of major blood loss.

Effects of hemoglobin-based blood substitutes on vasoactivity of rat aortic rings.

Our objective is to characterize the vasoactive properties of a 10% alphaalpha diaspirin cross-linked human hemoglobin (alphaalphaHb) and to test the hypothesis that sodium nitroprusside (SNP)-induced relaxation is inhibited in the presence of alphaalphaHb. Experiments were performed on aortic rings from 18 Wistar rats; the rings were suspended in aerated Krebs solution. Changes in isometric tension were measured to increasing concentrations of alphaalphaHb (1.8 x 10(-9) to 10(-4) M) on phenylephrine (PE)-induced contraction (3 x 10(-7) M), on acetylcholine (ACh)-induced relaxation (10(-8) to 10(-6) M), on SNP-induced relaxation (10(-9) and 10(-8) M), and on PE-induced contraction with an endothelin-1 (ET1) receptor antagonist, BQ123 (10(-5) M). Control rings received no alphaalphaHb. A concentration-dependent increase of the PE-precontraction (1.3%, 6.8%, 17.4%, and 34%, respectively) as well as the inhibition and reversal of ACh-induced relaxation was observed after alphaalphaHb. The presence of alphaalphaHb decreased the SNP-induced relaxation in the presence or absence of endothelium. The relaxation induced by SNP was reduced with time in the presence, but not in the absence, of alphaalphaHb. In conclusion, although pharmacological modulation of the vasoconstriction is possible with nitric oxide donors, our findings suggest that in the clinical setting, large sustained donor doses may be required.