Arterial flow regulator enables transplantation and growth of human fetal kidneys in rats.

Here we introduce a novel method of transplanting human fetal kidneys into adult rats. To overcome the technical challenges of fetal-to-adult organ transplantation, we devised an arterial flow regulator (AFR), consisting of a volume adjustable saline-filled cuff, which enables low-pressure human fetal kidneys to be transplanted into high-pressure adult rat hosts. By incrementally withdrawing saline from the AFR over time, blood flow entering the human fetal kidney was gradually increased until full blood flow was restored 30 days after transplantation. Human fetal kidneys were shown to dramatically increase in size and function. Moreover, rats which had all native renal mass removed 30 days after successful transplantation of the human fetal kidney were shown to have a mean survival time of 122 days compared to 3 days for control rats that underwent bilateral nephrectomy without a prior human fetal kidney transplant. These in vivo human fetal kidney models may serve as powerful platforms for drug testing and discovery.

Successful transcatheter closure of a large patent ductus venosus with the Amplatzer vascular plug II.

Patent ductus venosus is a rare form of congenital portosystemic shunt from the fetal umbilical vein to the inferior vena cava. The reported surgical treatments include ligation, banding, and liver transplantation. In addition, transcatheter closure with a coil, stent, or original Amplatzer vascular plug (AVP) has been reported. The AVP II, a redesigned version of the original vascular plug with a finer more densely woven nitinol wire and a large diameter (up to 22 mm) is available. This reported case is the first successful occlusion of a large patent ductus venosus with the new AVP II.

Ascending cholangitis provokes IL-8 and MCP-1 expression and promotes inflammatory cell infiltration in the cholestatic rat liver.

BACKGROUND/PURPOSE: Postoperative cholangitis is one of the most common complications after bile duct reconstruction. The pathogenesis and early consequences of ascending cholangitis still are unidentified. METHODS: Male Sprague-Dawley rats were divided into 5 treatment groups: control (n = 4), blood sampling and liver biopsy only; group I, [BDL/Eschericha coli; n = 6], ligation of common bile duct (BDL) for a week, followed by Roux-en-Y choledochojejunostomy (RYCJ) and injection of E coli (ATCC 25922) into Roux limb after 24 hours; group II, [BDL/NS; n = 5], same procedures as in group I, with injection of normal saline (NS) into Roux limb; group III, [SBDL/E coli; n = 6], primary RYCJ was constructed 1 week after sham ligation of common bile duct (SBDL) followed by the same treatment as group I; Group IV, [SBDL/N.S; n = 6], same procedures as in group III, but injecting NS into Roux limb. All animals were killed after 24 hours of treatment. Blood was sampled for culture and serum cytokine levels. The liver was harvested for quantitative bacterial culture, as well as for MCP-1, interleukin (IL)-8 (CINC in the rat) and transforming growth factor beta1 mRNA expression by reverse transcriptase polymerase chain reaction (RT-PCR) and for immunohistochemistry. The choledochojejunostomy was resected for culture. Serum cytokine levels were detected by ELISA kits. RESULTS: A significant increase of E coli ATCC 25922, occurred in the livers of group I rats, compared with group IV (P =.037). MCP-1 expression increased in all groups, compared with control (P =.000). The IL-8 mRNA expression was significantly higher in group I than in control (P =.021). The expression of TGF-beta1 mRNA was similar among the groups (P =.361), consistent with the immunohistochemistry results. The serum MCP-1 and IL-8 levels were higher in the 4 groups than in the control (P =.000) and were significantly higher in group I than in group IV (P =.001). CONCLUSIONS: This study found that a significant colonization of E coli of the same strain was present in the cholestatic rat liver injected into the Roux limb, which was associated with a higher expression of liver MCP-1 and IL-8 mRNA, a significant increase of serum MCP-1 and IL-8, and a more evident inflammatory cell infiltration into the porta hepatis.

Changes in bacterial concentration in the liver correlate with that in the hepaticojejunostomy after bile duct reconstruction: implication in the pathogenesis of postoperative cholangitis.

Postoperative cholangitis is a frequent and unpredictable complication of unknown etiology following bile duct reconstruction (BDR), particularly for biliary atresia. This study was undertaken to correlate the growth of bacteria in the hepaticojejunostomy with that in the liver after BDR. Quantitative bacterial culture was done on the specimens taken from the liver and from the hepaticojejunostomy at 1 week (group 1, n = 7), 1 month (group 2, n = 7), and 2 months (group 3, n = 7) following BDR with Roux-en-Y hepaticojejunostomy in piglets after 2 weeks of common bile duct ligation. The histological examination of the liver and the hepaticojejunostomy, as well as serial monitoring of hemogram and liver function tests, were performed to correlate the findings with the bacterial concentration of the liver and the hepaticojejunostomy following BDR. The bacterial concentration of the hepaticojejunostomy, expressed as log10 colony-forming units per gram (log10 CFU/g) of the hepaticojejunostomy, showed a progressive decrease from 8.38 +/- 1.36 in group 1, 7.07 +/- 2.54 in group 2, to 3.56 +/- 1.31 in group 3 (p = 0.001). The log10 CFU/g of the liver also showed a progressive decrease from 5.02 +/- 1.59 in group 1, 3.16 +/- 1.56 in group 2, to 2.19 +/- 1.09 in group 3 (p = 0.006). There was a significant positive correlation of the log10 CFU/g of the liver (n = 21) with that of the hepaticojejunostomy (n = 21) following BDR (r = 0.600, p = 0.004). Most of the infectious pathogens isolated from the liver were also isolated from the hepaticojejunostomy. The changes in hemoglobin, bilirubin, albumin, and ammonia significantly correlated with the changes of the bacterial concentration of the liver. The results of the study suggests that hepatic bacterial proliferation after BDR is significantly affected by microbial overgrowth in the bilioenteric anastomosis and is associated with deteriorated liver function and hemogram.

Reappraisal of the role of the bilioenteric conduit in the pathogenesis of postoperative cholangitis.

The incidence of postoperative cholangitis has changed very little despite progressive improvement in the treatment of biliary atresia. The role of the bilioenteric conduit in its pathogenesis is still uncertain. A retrospective study of 39 patients undergoing either a conventional Kasai operation (group 1, n = 20) or with placement of an antireflux valve (group 2, n = 10) or lengthening (group 3, n = 9) of the jejunal conduit from 40 to 60 cm was performed to compare the incidence of cholangitis. Postoperative cholangitis developed in 18 of the 39 patients (46%). The incidence was 10/20 (50%) in group 1, 5/10 (50%) in group 2, and 3/9 (33%) in group 3 (P = 0.679). An animal experiment was conducted concomitantly to compare quantitative bacterial cultures of the bilioenteric anastomosis and the liver before and 1 week after Roux-en-Y hepaticojejunostomy (HPJ) in piglets without (group A, 25 cm) and with (group B, 50 cm) lengthening of the jejunal conduit in a porcine model of extrahepatic biliary obstruction. The growth of bacteria in both the bilioenteric anastomosis and the liver was not affected by lengthening the jejunal conduit from 25 to 50 cm (P = 0.612 and 0. 057, respectively), despite a geometric increase in bacterial concentrations in both groups after HPJ. It is concluded that neither bacterial growth in the liver nor cholangitis following bile-duct reconstruction was affected by valving or lengthening the bilioenteric conduit.

Prompt colonization of the hepaticojejunostomy and translocation of bacteria to liver after bile duct reconstruction.

BACKGROUND/PURPOSE: Ascending cholangitis is one of the most common and earliest complications after bile duct reconstruction (BDR), particularly in patients with biliary atresia. To elucidate if the local factors such as the hepaticojejunostomy (HPJ) was responsible for the infection and the level of HPJ would affect the incidence of bacterial translocation (BT) to liver, this study was conducted. METHODS: Fourteen piglets received BDR 2 weeks after ligation of the common bile duct (CBDL). They were further divided into two groups according to the level of HPJ: group 1 (n=8), at the common hepatic duct and group 2 (n=6), at the bifurcation. One week after BDR, all the animals were killed. The liver and HPJ were harvested for quantitative tissue culture and for histological study. RESULTS: All HPJ were heavily colonized with at least one, mostly about two to four types of bacteria with a range of the quantitative bacterial growth from 3.9 x 10(4) to 8.6 x 10(9) colony-forming units (CFU)/g and a median of 8.3 x 10(6) CFU/g. Positive BT to liver, defined by more than 1 x 10(2) CFU/g, was found in 0 of the 14 piglets on day 1, 4 of 14 (29%) after CBDL, but in 9 of 14 (64%) after BDR (P=.001). Eight of the nine animals with positive BT to liver have the same species of bacteria found in their corresponding HPJ. Incidence of BT to liver was four of eight (50%) in group 1 and five of six (83%) in group 2, which was not significantly different (p=.30). CONCLUSIONS: This study suggests that there is prompt colonization of HPJ 1 week after BDR, which is in turn an important local factor responsible for BT to liver. The level of HPJ does not affect the incidence of BT to liver.

Adverse metabolic and microbiological effects of tube feeding in experimental canine obstructive jaundice.

BACKGROUND: Inadequate oral intake and poor absorption result in malnutrition in obstructive jaundice. Both malnutrition and obstructive jaundice promote bacterial translocation from the gut. This study was designed to test the efficacy of tube feeding in preventing malnutrition and in decreasing the metabolic and microbiological adverse effects in obstructive jaundice. METHODS: Forty adult mongrel dogs were studied and were allocated into one of four groups: group I (PO-control) underwent sham ligation of the common bile duct (CBDL) and was fed ad libitum on Portagen (Mead Johnson, Evansville, IN); group II (PO-CBDL) underwent CBDL and was ad libitum fed on the same formula; group III (FEG-control) underwent sham CBDL and received forced esophagogastric feeding (FEG) with Portagen; and group IV (FEG-CBDL) underwent CBDL and received FEG. All the animals underwent insertion of a F-12 feeding tube to the stomach from an esophagotomy wound on day 1 and the tube was used for continuous enteral feeding with Portagen over 4 h/d from day 2 until day 13 in groups III and IV. Fourteen days later, blood samplings were done and a laparotomy was performed to obtain liver, mesenteric lymph nodes (MLN), and terminal ileum for quantitative bacterial culture. Bacterial translocation to MLN and liver was represented by log10 CFU/g of tissue in this study. RESULTS: Both group II and IV animals with CBDL significantly lost body weight (p = .0001) and had a lower level of prealbumin (p = .0054). A significant increase in bacterial translocation to MLN and to liver occurred in groups II and IV (p = .0017 and .0268, respectively). Intestinal bacterial population was also higher in these two groups than in the other two controls (p = 0.0028). An increase in plasma ammonia level was found in dogs with CBDL (p = .0002) and in dogs with FEG (p = .003), compared with their respective controls. Three among 13 dogs in group IV died and no mortality occurred in the other groups (p = .223). CONCLUSIONS: Tube feeding fails to improve malnutrition in obstructive jaundice and is associated with intestinal bacterial overgrowth, promoting bacterial translocation to MLN and liver, precipitating liver dysfunction and consequently a higher mortality.

Metabolic effect of parenteral nutrition in dogs with obstructive jaundice.

OBJECTIVE: To assess the effect of total parenteral nutrition (TPN) on macronutrient metabolism in obstructive jaundice. METHODS: Forty adult mongrel dogs were equally divided into four groups: group I (PO-control) received sham ligation of common bile duct (CBDL) and was fed dog chow and water ad libitum; group II (PO-CBDL) underwent CBDL and was fed dog chow; group III (TPN-control) received sham CBDL and TPN; and group IV (TPN-CBDL) underwent CBDL and received TPN. Blood chemistries, plasma amino acids and liver histologies were studied before (Day 1) and at the end (Day 14) of the experiment. RESULTS: A significant elevation of bilirubin and alkaline phosphatase was observed in dogs with CBDL. Blood glucose was not changed significantly in any group. Significant increases in triglyceride and cholesterol were present in CBDL dogs. Significant differences in the concentrations of a few plasma amino acids, including an elevation of phenylalanine, were found in TPN dogs. A significant increase in aromatic amino acids (AAA) and a noticeable depression of the molar ratio of branched-chain amino acids (BCAA) to AAA was present in TPN-CBDL dogs, as was a significant increase in blood ammonia. CONCLUSION: In the presence of obstructive jaundice, TPN does not significantly affect carbohydrate or lipid metabolism. However, a derangement in protein metabolism with the standard TPN solution in current use suggests that either a modification of amino acid composition or an increase in total energy to protein energy ratio in TPN solution may be necessary to obviate such a consequence.

Role of parenteral nutrition in preventing malnutrition and decreasing bacterial translocation to liver in obstructive jaundice.

Surgery in patients with obstructive jaundice is associated with significant infectious complications probably due to impaired immune function and malnutrition. Total parenteral nutrition (TPN) may alleviate malnutrition but may also promote bacterial translocation (BT) from the gut. To elucidate if TPN can prevent malnutrition without promotion of BT in obstructive jaundice, 40 dogs underwent laparotomy for tissue sampling and placement of a central venous line and were allocated into one of four groups: I (PO-control) received dog chow and water ad libitum; II (PO-CBDL) underwent ligation of common bile duct (CBDL) and was fed dog chow; III (TPN-control) received TPN; and IV (TPN-CBDL) underwent CBDL and received TPN. Body weight, blood samples for liver function tests and bacterial culture, and tissues from liver and mesenteric lymph nodes (MLN) for quantitative bacterial culture and for histology were obtained prior to and 2 weeks after the experiment. The incidence of BT to MLN was 40% in the PO-CBDL and TPN-CBDL animals, which was significantly different from the other two groups (0%; p < 0.05). The incidence of BT to liver was 70% (7/10) in the PO-CBDL animals, which was significantly higher than that in groups I, III, and IV (0%, 20%, 20%, respectively) (p < 0.05). The PO-CBDL animals showed a significant decrease in body weight and prealbumin compatible with malnutrition, whereas the TPN-CBDL animals showed a significant increase in alkaline phosphatase and a consistent cholestasis on histology. The data suggest that TPN can prevent jaundice-associated malnutrition and decrease BT to liver but should be administered cautiously because it may precipitate cholestasis.

Nitrate Assimilation and Crassulacean Acid Metabolism in Leaves of Kalanchoë fedtschenkoi Variety Marginata.

The enzymes necessary to assimilate ammonia either via glutamine synthetase and glutamate synthase or via the glutamate dehydrogenase pathways are present in both green and white leaf tissues of Kalanchoë fedtschenkoi. Nitrate reductase activity develops to a maximum in a Crassulacean acid metabolism (CAM) plant canopy before either ribulose 1,5-bisphosphate carboxylase, or phosphoenolpyruvate carboxylase, or CAM. Nitrate reductase also is activated each morning and is inactivated late in the day as in other plants. However, there does not appear to be any direct relationship between nitrate reductase activity and the level of acid, its daily pattern or the amplitude of CAM. Though nitrate reductase is activated maximally each day by light, in Kalanchoë leaves for six days the activity followed a precise daily pattern independent of continuous light or dark.

Chronic industrial exposure to lead in 63 subjects. I. Clinical and erythrokinetic findings.

Clinical, biochemical, haematological and erythrokinetic studies were performed on 63 adult males with prolonged lead exposure. Their most common symptoms and findings were abdominal pain (62%), gingival lead lines (48%), headache and/or dizziness (33%), muscle cramps (32%), anaemia (19%), and fatigue (18%). Colicky abdominal pain (27%) and gingival lead lines correlated with urinary lead excretion. Anaemia was mild, but more frequent in the subjects with the greatest urinary lead excretion. Other associated findings were: higher reticulocyte counts and more basophilic stippling of the RBCs, more sideroblasts and greater erythroid hyperplasia of the bone marrow, more reduction in 51Cr-tagged RBC survival time, smaller RBC mass, a more rapid plasma iron clearance, a greater plasma iron turnover and greater utilization of 59Fe in subjects with urinary lead excretion of greater than 100 microng/day in comparison with the remainder and normal controls. These findings suggest that minimal chronic exposure to lead causes an increased haemolysis with resulting increased production of erythrocytes.

Chronic industrial exposure to lead in 63 subjects. II. Evaluation of chelation therapy.

Efficacy of chelation therapy with intravenous calcium disodium edetate, oral Ca EDTA, and oral penicillamine was tested in 63 subjects with chronic minimal industrial exposure to lead. All three agents increased the urinary lead excretion. The effect was greatest with intravenous Ca EDTA, next with oral penicillamine and least with oral Ca EDTA. Symptoms, particularly colicky abdominal pain, improved during the period of chelation therapy. Anaemic subjects showed improvements in haematological parameters. It is recommended that subjects with chronic minimal industrial exposure to lead receive chelation therapy. The relative merits of the three agents are discussed.

Hypersplenism in Wilson's disease.

Thirteen patients with Wilson's disease were compared with seven cirrhotic and 13 normal controls to define better the haematological abnormalities in this condition. Hypersplenism (anaemia, leukopenia, thrombocytopenia, and reduced red cell survival) commonly occurred in patients with both Wilson's disease and cirrhosis. These abnormalities correlated with splenic enlargement. Despite reduced haematocrits, red cell mass was greater in these two groups than in normal controls. Plasma volume and the body haematocrit/peripheral haematocrit ratios were also greater in patients with Wilson's disease and cirrhosis. Increased splenic sequestration of (51)Cr-tagged red blood cells was not demonstrated in any subjects. The hypersplenism in patients with Wilson's disease is similar to that found in patients with cirrhosis from other causes.