Correction of congenital heart defects in Jehovah's Witness children.

UNLABELLED: Between August 1989 and July 2003 14 Jehovah's Witness children with congenital heart defects (CHD) aged under 14 years (median 2.9 years) and with a median weight of 14 kg underwent 16 operations with cardiopulmonary bypass (CPB). Five children had been operated on previously between one to three times. Preoperatively, 7 children were prepared with oral iron supplementation and 10 received erythropoietin. Mean hemoglobin (Hb) at admission was 14.4 g/dl (range 10.9 - 19.2). The cardiopulmonary bypass (CPB) circuit was modified to reduce total priming volume. High doses of aprotinin were administered. The modified ultrafiltration (MUF) circuit, used in 7 patients, was parallel to the ECC circuit with continuous circulation of the blood through a small shunt between the arterial and venous lines. Operations performed consisted of VSD closure (3 pts.), ASD closure (3 pts.), Fontan operation (2 pts.), and complete AV canal correction, aortic commissurotomy, Ross operation, Glenn shunt, cor triatriatum correction, MV reconstruction combined with left outflow tract stenosis resection, correction of absent pulmonary valve syndrome, and correction of tetralogy of Fallot in one patient each. There were no deaths. Mean duration of CPB was 192 min and mean aortic cross-clamp time 40 min. The Hb value at the end of the operation was 4.9 - 14.5 g/dl (mean 9.6) and at discharge it was 7.1 - 14.5 g/dl (mean 15.5). No blood or blood products were used in any patient. CONCLUSION: Bloodless cardiac surgery with and without CPB can be safely performed in Jehovah's Witness infants and children.

Cerebral oxygen monitoring during neonatal cardiopulmonary bypass and deep hypothermic circulatory arrest.

BACKGROUND: This study was undertaken to investigate the physiological effects of cardiopulmonary bypass (CPB) and deep hypothermic circulatory arrest (DHCA) on cerebral oxygen metabolism estimated by near-infrared spectroscopy (NIRS). METHODS: Ten newborn piglets (2.1 to 2.6 kg) were monitored with right frontal NIRS; the right jugular bulb was cannulated for intermittent sampling of jugular venous blood. All animals underwent CPB, cooling to a core temperature below 15 degrees C, 60 minutes of DHCA followed by subsequent reperfusion and rewarming. Continuously recorded NIRS data and intermittent jugular venous blood values were compared. RESULTS: NIRS performance was examined over the jugular venous oxygen saturation (SjvO2) range of 40 to 98 %, a linear correlation was found between SjvO2 and NIRS-derived regional cerebral oxygen saturation (rSO2) (r = 0.91, p < 0.001). A correlation was observed between the cellular oxidation NIRS-parameter cytochrome oxidase aa3 (CytOx) slope during the DHCA period in relation to rectal and nasopharyngeal temperature immediately before the onset of DHCA (r = 0.75 and 0.85, p < 0.001). CONCLUSIONS: This study suggests that NIRS-measured hemoglobin oxygenation parameters may reflect functional changes in cerebral hemodynamics and brain tissue oxygenation, while CytOx values represent related effects on intracellular oxidative metabolism.

The measurement of muscle protein synthesis in broilers with a flooding dose technique: use of 15N-labelled phenylalanine, GC-MS and GC-C-IRMS.

An experiment was carried out to measure fractional muscle protein synthesis rates (k(s)) in broilers with injection of a flooding dose of phenylalanine (1 ml/100 g body weight of 150 mM phenylalanine; 38 atom percent excess (APE) [15N]phenylalanine). K(s) was calculated from the [15N] enrichment in phenylalanine of tissue-free and protein-bound phenylalanine using both gas chromatography mass spectrometry (GC-MS) and gas chromatography combustion isotope ratio mass spectrometry (GC-C-IRMS) for measurements after a 10 min isotope incorporation period. The tertiary-butyldimethylsilyl (t-BDMS) derivatives of phenylalanine were used for gas chromatographic separation in both systems. GC-MS and GC-C-IRMS were calibrated for a range of 7 to 37 [15N]APE and 0 to 0.62 [15N]APE, respectively, and for sample sizes of 0.45 to 4.5 nmol phenylalanine and 7 to 40 nmol phenylalanine, respectively. Reproducibility of standards as a measure of precision varied from 0.06 to 0.29 [15N]APE and from 0.0004 to 0.0018 [15N]APE in GC-MS and GC-C-IRMS, respectively. K(s) was measured in the m. pectoralis major of broilers fed rye based diets (56%) which were provided either unsupplemented (-) or supplemented (+) with an enzyme preparation containing xylanase. K(s) in breast muscles was significantly increased from 21.8%/d to 23.9%/d due to enzyme supplementation. It can be concluded from the study that the measurement of protein synthesis in broilers with the flooding dose technique can be carried out by using [15N]phenylalanine, GC-MS and GC-C-IRMS.

Improved myocardial preservation with short hyperthermia prior to cold cardioplegic ischemia in immature rabbit hearts.

OBJECTIVE: Recent observations have been shown that the induction and accumulation of heat shock proteins (HSPs) by short exposure to nonlethal whole-body hyperthermia with normothermic recovery are closely associated with transient resistance to subsequent ischemia-reperfusion challanges. Here, this study was performed to investigate whether a shortly heat shock pretreatment affects the left ventricular (LV) function after cold cardioplegic ischemia in reperfused neonatal rabbit hearts. METHODS: Hearts from neonatal New Zealand White rabbits were isolated perfused (working heart preparation) and exposed to 2 h of cold cardioplegic ischemia followed by reperfusion for 60 min. To induce the heat shock response neonatal rabbits (n=5, HT-group) were subjected to whole-body hyperthermia at 42.0-42.5 degrees C for 15 min, followed by a normothermic recovery period of 60 min, before harvesting and the onset of global hypothermic cardioplegic arrest. Another set of hearts (n=5, control group) without a heat treatment underwent a similar perfusion and ischemia protocol served as control. The postischemic recovery was assessed by measuring several parameters of LV function. LV biopsies from all control and heat treated animals were taken before ischemia and at the end of reperfusion to examine myocardial HSP levels by Western blot analysis. RESULTS: At 60 min of reperfusion the HT-group showed significant better recovery of ventricular function such as LV developed pressure (DP) (74.6+/-10 vs. 52.1+/-8.5%, P<0.05), LV positive dP/dt (910+/-170 vs. 530+/-58 mmHg/s, P<0.01) and LV end-diastolic pressure (LVEDP) (8+/-2 vs. 18.4+/-5 mmHg, P<0.05) than control. Myocardial oxygen consumption (MVO(2)) was significantly higher in the HT-group compared with control (0.054+/-0.006 vs. 0.041+/-0.002 ml/g per min, P<0.05). Significant postreperfusion lower level in lactate production was observed in the HT-group (0.83+/-0.11 vs. 1.67+/-0.8 mmol/l, P<0.05). Also, the recovery of hemodynamic parameters such as aortic flow, coronary flow and cardiac output was significantly superior (P<0.05) in the HT-group. Furthermore, high expression of HSP72(+)/73(+) were detected in the myocardial tissue samples of heat-treated rabbits by immunoblotting, appearing even at 60 min of normothermic recovery after heat stress. CONCLUSIONS: These data in the immature rabbit heart indicate that previous shortly heat treatment with high level expression of heat shock proteins (HSP72(+)/73(+)) before hypothermic cardioplegic ischemia provides transient tolerance against myocardial injury and could be an improvement for the postischemic functional recovery of neonatal hearts.

Replacement of soybean oil with tallow in rye-based diets without xylanase increases protein synthesis in small intestine of broilers.

We examined the effects of dietary fat type (10% of either soybean oil, S, or beef tallow, T)(3) and xylanase supplementation (-, without; +, with 1 g of Avizyme 1300 per kg diet) in rye- based diets (56%) on tissue protein synthesis in male broilers. Birds were injected with a large flooding dose of a phenylalanine solution (150 mmol/L, 38 atom percentage excess [(15) N] phenylalanine) and tissues were obtained after a 10-min incorporation period. [(15) N]-enrichment in tissue free phenylalanine and tissue protein bound phenylalanine were measured by gas-chromatography mass-spectrometry and by gas-chromatography combustion isotope ratio mass-spectrometry, respectively in order to calculate tissue specific fractional rates of protein synthesis (k(s)). The k(s) (%/d) in (S-), (S+), (T-) and (T+)-fed birds were 56, 64, 84 and 61 (SEM = 3.7) in duodenum, 51, 52, 75 and 58 in jejunum (SEM = 3.1), 66, 67, 105 and 68 (SEM =7.0) in jejunal mucosa cells, 53, 56, 68 and 50 (SEM = 3.7) in ileum and 52, 45, 118 and 39 (SEM = 20.2) in pancreas, respectively. Significant fat, enzyme or interaction effects in these tissues were mainly caused by the elevated k(s) in (T-)-fed birds which was closely associated with intestinal viscosity. We conclude that the effect of soluble nonstarch polysaccharides (NSP) and of NSP-hydrolyzing enzymes may be explained partially by modification in tissue protein synthesis of the intestinal tract.

Protein S-100beta in brain and serum after deep hypothermic circulatory arrest in rabbits: relationship to perivascular astrocytic swelling.

The aim of this study was to evaluate the relationship between the kinetic patterns of the protein S-100beta, an astroglial cell marker, and its immunohistochemical expression in the brain in rabbits that underwent cardiopulmonary bypass with deep hypothermic circulatory arrest. Fourteen New Zealand rabbits (weight, 3.1+/-0.25 kg) were anaesthetised, intubated and mechanically ventilated. Four animals were not connected to the cardiopulmonary bypass and served as controls. Ten animals were perfused according to a uniform protocol. After systemic cooling, deep hypothermic circulatory arrest was induced for 60 minutes. After reperfusion and rewarming, the animals were weaned from bypass and sacrificed. In the brain, astrocyte reactivity for S-100beta was evaluated immunocytochemically (DPC Immustain) and the serum concentrations of S-100beta were analysed using a commercially available immunoluminometric kit (Byk-Sangtec, Dietzenbach, Germany). In all experimental animals a significant increase of the serum concentration of the protein S-100beta was found immediately after reperfusion and the termination of cardiopulmonary bypass. In comparison with the control animals, increased staining of S-100beta was found in the astroglial cells and swollen astrocytic end-feet in the perivascular regions. There were fewer signs of neuronal cell injury of neurones in the hippocampus structure. In conclusion, astrocytic activation and S-100beta overexpression seems to precede the neurodegeneration following deep hypothermic circulatory arrest. The marked perivascular cell swelling may support the assumption of reperfusion injury of the astroglial cell complex that forms the blood-brain barrier, which may be indicative of the source of the released S-100beta into the blood stream.

The effect of continuous treatment with sodium nitroprusside on the serum kinetics of the brain marker protein S-100beta in neonates undergoing corrective cardiac surgery by means of hypothermic cardiopulmonary bypass.

Neonates who undergo cardiac surgery of d-transposition of the great arteries by means of hypothermic cardiopulmonary bypass (CPB) represent a group at increased risk to develop brain injury and altered psychomotor development in early life. Measurement of protein S-100beta, an astrocytic calcium binding protein, in serum may provide information on transient astroglial cell activation and disintegration of the related blood-brain barrier due to oxidative stress during and after CPB. Conflicting results have been reported that concern the neuroprotective effect of the NO liberator sodium nitroprusside (SNP) in vitro. We evaluated the effect of continuous treatment with SNP on the serum kinetics of S-100beta in infants and children after corrective cardiac surgery. The data on 25 neonates treated intraoperatively and postoperatively and 28 without treatment were analyzed. SNP was infused (1-5 microg/kg body weight/minute depending on the haemodynamic status) after the induction of anesthesia, and during and after the termination of CPB for 2 days. Serum concentrations of S-100beta were analyzed by the use of a commercially available immunoluminometric kit (Byk-Sangtec, Dietzenbach, Germany). There were no significant differences in the bypass data between the SNP-treated and non-treated group. In comparison to the pre-bypass values, a similar increase in the concentration of protein S-100beta was found 2 hours after the termination of CPB in the SNP-treated and non-treated neonates, which decreased over the subsequent 48 postoperative hours. However, significantly lower post-bypass serum levels of S-100beta were found in the SNP-treated group after 24 hours (p = 0.0009) and 48 hours (p = 0.04) of treatment. In conclusion, the significant elevation of serum levels of protein S-100beta may indicate increased astroglial cell reactivity and increased passage into the blood stream. Longer-lasting treatment with NO liberator SNP seemed to decrease the release of S-100beta into the blood stream and may have delayed protection on the astroglial cells. The neurological significance of such an observation, however, should be evaluated in further follow-up studies, which need to include additional neurophysiological and neurodevelopmental tests.

Responses in the absorptive phase in muscle and liver protein synthesis rates of growing rats.

The effect of time after beginning of a meal (30, 60, 90 and 120 min) on liver and gastrocnemius muscle protein synthesis was tested in growing male rats using the large dose technique, based on a 10 min exposure to [15N]phenylalanine. The fractional synthesis rate was estimated from the ratio between the atom percent excess of tissue protein-bound and free labelled phenylalanine. The latter was measured by gas chromatography mass spectrometry using the tertiary-butyldimethylsilyl amino acid derivatives. The protein-bound phenylalanine of gastrocnemius muscle was separated from the other amino acids using preparative amino acid chromatography and then oxidised to N2 in an automated carbon-nitrogen Roboprep (CN) combustion module attached to a continuous flow isotope ratio mass spectrometer (IRMS), with m/z ions 28 and 29 monitored. The protein-bound phenylalanine from liver was separated by a gas chromatograph attached to a sample preparation module and an isotope ratio mass spectrometer (GC C-IRMS), with again m/z ions of 28 and 29 monitored. The following results were obtained: the daily fractional protein synthesis rates (ks) in gastrocnemius muscle and liver were 13.9% and 65.6% respectively, in 12 h fasted 145 g rats. These ks increased within 30 min after ingestion of meal to 14.9% and 91.8% for muscle and liver, respectively, and remained at these values for the next 90 min (14.6% and 87.4% at 60 min, and 14.3% and 88.6% at 120 min after the beginning of feeding). It was concluded that measurement of protein synthesis rates characteristics for the absorptive phase can be undertaken in a period from thirty minutes to two hours after start of a meal, without significant changes in the ks values.

Quantitative and functional restorations and alterations of peripheral lymphocytes in patients with autologous spleen implantation.

To reduce the risk of severe infections in splenectomized patients, new methods for splenic preservation or heterotopic autologous spleen implantation have been established. In the latter case, the immunological and functional benefits are still under discussion. In this study we compared immunological parameters in 16 splenectomized patients with and without heterotopic autologous spleen implantation with a nonsplenectomized control group. The total lymphocyte counts--T-cells, CD4+ -, as well as CD8+ - lymphocytes, CD16+ - and B-cells--were highly elevated in both groups, whereby the B-cells were relatively and absolutely higher in the implanted group than in the nonimplanted group. Splenectomized patients had a significantly reduced serum IgM level. The serum IgM of patients with splenic autotransplantation was not significantly lower than that of the controls. In contrast to the impaired in vitro immunoglobulin synthesis in the splenectomized group, the autotransplanted patients showed a normal PWM-induced IgG and IgM synthesis and an increased IgA production compared with the controls. The latter results support the findings of elevated serum IgA levels in this group. The mitogenic-induced proliferation with PHA, ConA, PWM, and OKT3 was not clearly different within the tested groups. The results may indicate a benefit of autologous spleen implantation in regard to the humoral immune response.

Neurotensin-like immunoreactivity released into the portal vein by duodenal acidification in the dog.

The present study in the dog evaluates neurotensin as a potential hormone, mediating the inhibition of gastric acid secretion by duodenal acidification. Histamine-stimulated acid output was determined before and during duodenal acidification. Portal vein blood was obtained and assayed for carboxy-terminal neurotensin-like immunoreactivity (NTLI). Duodenal perfusion with 15 mmol HCL for 30 min significantly inhibited histamine-stimulated acid output to 67% of control output. This inhibition was not associated with any change in the peripheral plasma NTLI, but the portal plasma NTLI was significantly elevated from 27 to 78 pM. The effect of duodenal acidification on liver extract meal-stimulated acid secretion was determined in a second group of dogs without portal vein catheter. Duodenal perfusion with 15 mmol HCl for 30 min significantly inhibited meal-stimulated acid secretion to 37% of control output. Intravenous infusion of synthetic neurotensin to a plasma level of 130 pM was required to inhibit meal-stimulated acid output significantly. In summary, NTLI is elevated in portal, but not peripheral, plasma after duodenal acidification. The associated inhibition of acid secretion is not due to hormonal action of neurotensin.

Molecular heterogeneity of canine cholecystokinin in portal and peripheral plasma.

The release of molecular forms of cholecystokinin (CCK) into the portal and peripheral blood in response to an intraduodenal perfusion of sodium oleate (9 mmol X h-1) was studied in six conscious dogs with chronic portal vein catheters. Immunoreactive CCK as concentrated from 20 ml plasma by C18 SEP PAK cartridges and the pattern of molecular forms of CCK were studied by G50 gel filtration. CCK-like immunoreactivity (CCK-LI) was measured in the column eluates with antibody 5135, which measures gastrin and CCK equally and requires the intact carboxyl-terminus for full recognition. Gastrin was measured specifically with antibody 1611. Intraduodenal perfusion with oleate did not alter basal gastrin release. Release of CCK-LI by intraduodenal oleate was calculated by the increments of the integrated CCK-LI peaks over basal. Total CCK-like immunoreactivity (CCK-LI), calculated by integration of all CCK-LI peaks in gel filtration eluates, increased over basal by 12 fmol/ml in the portal and by 6 fmol/ml in the peripheral plasma after intraduodenal perfusion with sodium oleate. The main molecular forms eluted on gel filtration in positions of CCK33,39 and of CCK8. The pattern of CCK in the peripheral plasma was similar to that in the portal plasma except that in the peripheral plasma large molecular forms were more abundant than small forms. This finding was confirmed when CCK39 and CCK8 were infused either into the portal vein or into the peripheral vein and peripheral plasma CCK levels were measured. Elimination of CCK8 after portal vein infusion compared to peripheral vein infusion was about 3 times higher than that of CCK39. The abundance of large molecular forms of CCK in the circulating blood which are similar in potency to small forms, underlines their role in the physiology of CCK.

[Perioperative antibiotic prophylaxis in elective splenectomy. A randomized prospective study]. / Perioperative Antibioticaprophylaxe bei der elektiven Splenektomie. Eine randomisierte, prospektive Studie.

After elective splenectomy septic complications in the early postoperative period occur up to 45%. In a prospective, randomised, controlled study of 61 patients the incidence of postoperative infections with and without prophylactic use of an antibiotic was compared. 61 patients undergoing elective splenectomy were divided into two groups. 28 patients were operated without any antibiotic, 33 patients were given 3 X 1500 mg Cefuroxim (Zinazef) for three days, starting 1 h prior to the operation. Infections of the urinary tract and the lung were excluded preoperatively. With a standardized program we searched for intraabdominal abscesses, infections of the lung, the urinary system and the abdominal incision postoperatively. No fatal complication occurred. A significant reduction in total septic complication rate (p less than 0.01) and in pneumonia was found as well in benign as in malignant disease. The study shows, that the apparently disturbed immunological defense following splenectomy can be improved by the prophylactic use of antibiotics.

Lack of correlation between mucus gel thickness and gastric cytoprotection in rats.

The effect of various cytoprotective agents on the thickness of gastric mucus gel layer in rats was studied. It was hypothesized that an increase in the mucus gel layer might be involved in cytoprotection. The results show that this is not the case. Neither prostaglandin E2, 16,16-dimethyl prostaglandin E2, nor mild irritants (20% ethanol, 0.35 M HCl, 20% glucose, 20% mannitol), all given orally, altered the thickness of the mucus gel layer, although these agents were found to be cytoprotective, i.e., inhibiting the formation of gastric mucosal necrotic lesions caused by oral administration of absolute ethanol. The only agents that significantly increased the thickness of the mucus gel layer were a hypertonic solution (4% NaCl) and sodium salicylate. We conclude that if mucus plays a role in cytoprotection, it is not by virtue of an increase in thickness of the gel layer adherent to the gastric mucosa.

[Biochemical parameters of hepatic ischemia in experimental animals]. / Biochemische Parameter der Leberischämie im Tierexperiment.

It was the aim of this study to define biochemical parameters of hepatic ischemia. In seven dogs portal vein and hepatic artery were occluded subsequently. During this ischemia the lactate concentration in the hepatic veins rose from a basal value of 15 mg% to 57 mg%. Lactate levels measured in portal vein, femoral artery, and femoral vein were significantly lower. Furthermore, hepatic ischemia caused an increase in potassium and glucose levels in the hepatic vein. Primarily, production of lactate by the liver seems to be an appropriate parameter for hepatic ischemia. Measuring lactate metabolism could be used to examine patients with portal hypertension before performing a shunt procedure. By cannulating the umbilical vein it is possible to occlude the portal vein with a balloon catheter. Portal vein occlusion with hepatic vein blood sampling may identify those patients in whom hepatic arterial reserve is inadequate to support portosystemic shunting.

[Conjugated cholic acid in the portal vein, aorta and hepatic vein in patients with alcohol toxic liver cirrhosis]. / Konjugierte Cholsäure in Pfortader, Aorta und Lebervene bei Patienten mit alkoholtoxischer Leberzirrhose.

In 12 patients with Laennec's cirrhosis conjugated cholic acid was measured by radioimmunoassay simultaneously in the portal vein, the aorta, and the hepatic vein. Furthermore, the concentration was measured for 90 minutes after i. v. injection of cholecystokinin. In the fasting patient the porto-venous extraction ratio was 0.45 (SD 0.23) and the arterio-venous extraction ratio was 0.24 (SD 0,21). 15-30 minutes after cholecystokinin the bile acid concentration significantly increased. In this time the porto-venous extraction ratio rose to 0.71 while the aorto-venous extraction ratio was different. These results agree with the hemodynamics found in cirrhosis. After cholecystokinin the increase in the extraction ratio account for the blood loss by porto venous shunts which corresponds to an increase of the portal compartment.