Histometric changes in the liver of inbred rats during prevention of hypokinesis]

An experimental investigation on morphological changes in the liver under prophylaxis of hypokinesia has been performed in genetically uniform material--150 premature rats of August and Wistar lines. The prophylaxis has been carried out at hypokinesia as the background produced by training in the treadban and at the combination of the latter with a pharmacological stimulator. The experiment has been performed for two months. In histological preparations of the liver the number of hepatic lobules, hepatocytes and stellate reticuloendotheliocytes per unit of the tissue area have been determined. Nuclear diameters, width of hepatocytes and sinusoid blood capillaries have been measured. Significant histometric changes in structural components of the liver have been stated after 60 days of hypokinesia with decrease of the organ. Prophylactic measures applied with hypokinesia as the background produce certain positive effect, though it is not the same in different indices and in different strains of animals.     

Effect of bile on vitamin B12 absorption.

The standard double-isotope Schilling test was used to study vitamin B12 absorption in seven patients with obstructive jaundice and 10 with T-tube bile duct drainage after cholecystectomy and bile duct exploration. In three and five of these patients respectively absorption was impaired. In the second group six patients were restudied after removal of the T tube, and in each case absorption was improved. Similar results were obtained after bile duct ligation in rats. Bile exclusion produced a 50-60% reduction in renal and hepatic uptake of vitamin B12 from the intestinal lumen. The malabsorption was corrected by replacing bile. These studies suggest that bile plays a part in the normal absorption of vitamin B12.     

Granular cell tumor of the biliary system: a report of 2 cases with cytologic diagnosis on endoscopic brushing

BACKGROUND: Granular cell tumors (GCTs) of biliary system are rare. GCTs show a striking preponderance for young, black females, who generally present with obstructive jaundice. To our knowledge, these are the first 2 reports of GCT of biliary system identifed on endoscopic brushing cytology. CASES: In case 1, a 24-year-old, black woman presented with a 5-month history of pruritus. Radiographic studies demonstrated a mass in the distal common bile duct. Endoscopic biopsy and bile duct brushing were diagnosed as GCT. A Whipple procedure was confirmatory of GCT. In case 2, a 38-year-old, black female presented with a 7-month history of pruritus and jaundice. Radiographic studies showed a stricture of the common hepatic duct at the hilum. Endoscopic brushing cytology of the stricture yielded only a few sheets of granular cells that were missed on initial screening. Suspicion of cholangiocarcinoma prompted surgery, and final histopathology showed GCT. Both patients were well 1 1/2 and 6 years after presentation. CONCLUSION: GCT of the bile duct can be diagnosed on endoscopic brushing and should be considered in the cytologic differential diagnosis in the appropriate clinical settings.     

Morphologic and histochemical indices of hepatic function following bilateral subdiaphragmatic vagotomy

A complex morpho-functional investigation of the rat liver was performed after bilateral subdiaphragmal vagotomy by means of light, electron microscopy methods and a quantitative histochemical method. Some ultrastructural disorders in hepatocytes and in stellate reticuloendotheliocytes were revealed, with their maximal manifestation 7 days after vagotomy. At later stages (45 and 90 days), compensatory-restorative processes develop resulting in considerable (but not complete) normalization of the organ's structure. Quantitative histochemical investigations have demonstrated that even at the highest degree of the resulted disorders the liver preserves its ability to synthesize and accumulate glycogen, but the intensity of the process is considerably lowered. Functional changes are reversible in their character and correlate to the degree of structural disorders.     

Changes in the lipid-accumulating cells of the liver during experimental hypervitaminosis A

Changes of the fat-storing cells (FSC) in the liver have been studied after oil solution of vitamin A have been injected to intact rats, when Kupffer's cells have been blocked with colloid silver and stimulated with prodigiosan. During these experiments (on the 4th, 7th, 14th days) the amount of FSC does not change, and their size increases. The blockade of Kupffer's cells does not influence the dynamics of the FSC changes, and their stimulation contributes to lipid accumulation in their cytoplasm and to earlier revealing of these elements (on the 2d day of the experiment). In parallel with increasing size of FSC, fatty infiltration of hepatocytes disappears and reticular fibers accumulate in the perisinusoid space. Participation of FSC in lipid metabolism and in synthesis of the III type collagen is proved, as well as existence of intercellular interactions of these elements with endotheliocytes, Kupffer's cells and hepatocytes.     

The role of bile acids in the reduction in lipopolysaccharide uptake by cultured rat Kupffer cells

The influence of bile salts on the binding and uptake of Salmonella abortus equi lipopolysaccharide by cultured Kupffer cells was studied. In control preparations, the percentage of cell-associated lipopolysaccharide increased with time and reached a plateau after about 2 h incubation at 37 degrees C. About 1.2 micrograms lipopolysaccharide was associated with 10(6) Kupffer cells at this time interval. In the presence of 0.3, 0.6 and 1 mumol bile salts/ml the cell-associated lipopolysaccharide was respectively, about 5%, 13% and 29% lower than in control cultures. In the presence of 1 mumol bile salts/ml, the association of lipopolysaccharide to cells at 0 degrees C was about 25% lower than in controls. Preincubation of Kupffer cells with 1 mumol bile salts/ml, with or without lipopolysaccharide, did not affect cell-associated lipopolysaccharide after removal of the bile salts. The rate of secretion of radioactivity by Kupffer cells was not influenced by the presence of bile salts during the uptake or the secretion periods. Bile acids proved to inactivate lipopolysaccharide. From these observations it was concluded that low concentrations of bile salts influence the binding and uptake of lipopolysaccharide by Kupffer cells. It was, therefore, considered likely that, in patients with obstructive jaundice, the high serum bile acid level accounts for spill-over of portal lipopolysaccharide into the systemic blood.     

Decreased collagen types I and IV, laminin, CK-19 and α-SMA expression after bone marrow cell transplantation in rats with liver fibrosis

Bone marrow cells have frequently been tested in animal models of liver fibrosis to assess their role in hepatic regeneration. The mononuclear fraction of bone marrow cells is of particular interest, as many studies show that these cells may be beneficial to treat hepatic fibrosis. In this study, we used the bile duct ligation model to induce hepatic fibrosis in an irreversible manner, and rats were treated with bone marrow mononuclear (BMMN) cells after fibrosis was established. Analysis of collagen types I and IV, laminin and α-SMA showed a decreased expression of these proteins in fibrotic livers after 7 days of BMMN cell injection. Moreover, cytokeratin-19 analysis showed a reduction in bile ducts in the BMMN cell-treated group. These results were accompanied by ameliorated levels of hepatic enzymes GPT (Glutamic-pyruvic transaminase), GOT (glutamic-oxaloacetic transaminase) and alkaline phosphatase (AP). Therefore, we showed that BMMN cells decrease hepatic fibrosis by significantly reducing myofibroblast numbers and through reduction of the collagen and laminin-rich extracellular matrix of fibrotic septa and hepatic sinusoids.     

Effects of bile acids and endotoxin on the function and morphology of cultured hamster Kupffer cells

The mechanisms of hepatic reticuloendothelial cell dysfunction in obstructive jaundice were investigated using cultured hamster Kupffer cells. The introduction of free bile acids, cholic acid (CA) at concentrations over 2 mM and chenodeoxycholic acid (CDCA) over 1 mM inhibited colloidal carbon pinocytosis. CA and CDCA at concentrations over 0.5 mM inhibited IgG-coated sheep red blood cell phagocytosis. With the application of conjugated bile acid and endotoxin at concentrations over 50 micrograms/ml, endocytic function was inhibited. With bile acids, a dose-dependent increase in the concentration of beta-glucuronidase occurred in the culture medium, and with endotoxin a time-dependent increase in beta-glucuronidase was noted. Bile acids produced alterations in cell organelles before destruction of the cell membrane. The presence of endotoxin led to the appearance of large vacuoles in the cytoplasm. These observations suggest that bile acids and endotoxin inhibit Kupffer cells by different mechanisms. We tentatively conclude that bile acids rather than endotoxin influence Kupffer cells in vivo.     

Bile acids in the rat: studies in experimental occlusion of the bile duct

Bile acids in the plasma, urine, and small intestine of adult male rats with occluded bile ducts have been studied using a method of high specificity for their determination. After bile duct ligation cholic acid rapidly accumulates in the plasma for 8 hr, remains high for a further 8 hr, and subsequently diminishes; bile acids disappear from the small intestine. During the first 12 hr after bile duct ligation the excretion of trihydroxy acids in the urine was 10 times that of the dihydroxy acids. Subsequently the two excretion rates became equal. Because bile acids have been implicated in the etiology of hepatic damage following bile duct ligation, studies have been made of the effect on the liver of removing (with cholestyramine) and supplementing (with cholic acid) the intestinal bile acid pool. The addition of cholestyramine to the stock diet prevented the rise in trihydroxy bile acids after bile duct ligation, but did not prevent the development of histological abnormalities in the liver. Supplementing the diet with cholic acid raised the plasma cholic acid levels but had little effect on the hepatic histological findings.     

Cytokine characteristics of jaundice in mouse liver

OBJECTIVE: The aim of this study is to clarify the perioperative cytokine changes and their mechanism in jaundiced liver. MATERIALS AND METHODS: Obstructive jaundice was induced using a common bile duct ligation (CBDL) and a two-thirds hepatectomy (HEP) was performed in six- to seven-week-old male C3H/HeN mice. When hepatectomy was added to CBDL, it was carried out 2 to 5 days after CBDL. The serum interleukin 6 (IL-6) levels and heat shock protein (HSP)-70 expression were evaluated. One mg per mouse of methylprednisolone (MPL) was intraperitonealy administered in some mice of CBDL+HEP group. RESULTS: The post-hepatectomy IL-6 values at 2 and 3 days after CBDL were significantly lower than those in the HEP group, while those at 5 days after CBDL were significantly higher than those in HEP group. The serum IL-6 value of the steroid group was significantly lower than that of non-steroid group in HEP group. However, no steroid effects were recognized on post-hepatectomy IL-6 values at 3 days after CBDL, steroid inhibited post-hepatectomy IL-6 production at 5 days after CBDL. No expression of HSP70 protein was observed in the control group, but HSP70 protein was expressed in both the hepatocytes and Kupffer cells 3 days after CBDL, then at 5 days after CBDL, no HSP70 protein was expressed in the Kupffer cells. CONCLUSIONS: In the early period of obstructive jaundice, the IL-6 level after hepatectomy did not increase in comparison to HEP group, and steroid had no effect on IL-6 level. According to the progression of obstructive jaundice, the IL-6 level after hepatectomy increased to a higher level than before, and the effect of MPL was restored. HSP70 is thus suggested to have an important role in cytokine production.     

Bone marrow transplantation results in donor-derived hepatocytes in an animal model of inherited cholestatic liver disease

Cell transplantation is a potential therapy for acquired or inherited liver diseases. Donor-derived hepatocytes (DDH) have been found in humans and mice after bone marrow transplantation (BMT) but with highly variable frequencies in different disease models. To test the effect of liver repopulation after BMT in inherited cholestatic liver diseases, spgp (sister of P-glycoprotein, or bile salt export pump, abcb11) knockout mice, a model for human progressive intrahepatic cholestasis type 2 with defects in excreting bile salts across the hepatocyte canalicular membrane, were transplanted with bone marrow cells from enhanced green fluorescent protein (EGFP) transgenic donor mice after lethal irradiation. One to 6 months later, scattered EGFP-positive DDHs with positive spgp staining were observed in the liver. These hepatocytes had been incorporated into hepatic plates and stained positively with hepatocyte-specific marker albumin. RT-PCR for the spgp gene revealed positive expression in the liver of sgsp knockout mice that had received the transplant. Bile acid analysis of bile samples showed that these mice also had higher levels of total biliary bile acid and taurocholic acid concentration than knockout mice without transplantation, indicating that BMT partially improved biliary bile acid secretion. Our results indicate that bone marrow cells could serve as a potential source for restoration of hepatic functions in chronic metabolic liver disease.     

Common bile duct obstruction caused by the hydatid daughter cysts

Echinococcosis is a human parasitary disease. In 2002, 29 new cases of liver echinococcosis were recorded in Croatia. Liver is the most common site of hydatid cysts. Nine patients with echinoccocal liver disease were operated in our department in 2002. Here we present a case where a patient with verified hydatid cyst in the left liver lobe developed high fever, jaundice, nausea, vomiting and pain in the upper abdomen. The symptoms were initially ascribed to the acute cholangitis. After unsuccessful antibiotic treatment, computerized tomography and endoscopic retrograde cholangiopancreatography (ERCP) were performed, demonstrating daughter cysts in the common bile duct. During ERCP, papilotomy was made and daughter cysts were extracted. Hydatid cyst was surgically removed, and a communication between the cyst and left hepatic duct was noted during surgery. Pericystectomy, choledochotomy, removal of remaining daughter cysts from the common bile duct, and sutures of left hepatic duct were performed. The patient recovered fully after the surgery. One of the possible complications of the liver hydatid cysts is the communication between cyst and the biliary tree. Such communications are usually asymptomatic, but symptoms can also mimic acute cholangitis and jaundice, which may lead to the misdiagnosis of the patient's condition.     

Effects of ionizing radiation on the activity of the major hepatic enzymes implicated in bile acid biosynthesis in the rat

In the days following high-dose radiation exposure, damage to small intestinal mucosa is aggravated by changes in the bile acid pool reaching the gut. Intestinal bile acid malabsorption, as described classically, may be associated with altered hepatic bile acid biosynthesis, which was the objective of this work. The activity of the main rate-limiting enzymes implicated in the bile acid biosynthesis were evaluated in the days following an 8-Gy gamma(60)Co total body irradiation of rats, with concomitant determination of biliary bile acid profiles and intestinal bile acid content. Modifications of biliary bile acid profiles, observed as early as the first post-irradiation day, were most marked at the third and fourth day, and resulted in an increased hydrophobicity index. In parallel, the intestinal bile acids' content was enhanced and hepatic enzymatic activities leading to bile acids were changed. A marked increase of sterol 12 alpha-hydroxylase and decrease of oxysterol 7 alpha-hydroxylase activity was observed at day 3, whereas both cholesterol 7 alpha-hydroxylase and oxysterol 7 alpha-hydroxylase activities were decreased at day 4 after irradiation. These results show, for the first time, radiation-induced modifications of hepatic enzymatic activities implicated in bile acid biosynthesis and suggest that they are mainly a consequence of radiation-altered intestinal absorption, which induces a physiological response of the enterohepatic bile acid recirculation.     

Liver Transplantation in Man—III,Studies of Liver Function,Histology, and Immunosuppressive Therapy

The experience gained from 13 hepatic transplant operations is described, with particular reference to the findings in nine patients who survived the immediate operative period. A major problem was found to be infection. Fulminant pneumonia caused death in two adults, at a time when liver function was virtually normal. Infection related to bile fistula and sepsis may be overcome by an improved method of biliary drainage by cholecyst-dochostomy, which was carried out in the last two patients. Jaundice in the second week due to rejection was observed in several patients. The striking histological change was centrilobular cholestasis. The jaundice, which was not prevented by administration of antilymphocyte globulin, was rapidly controlled by temporarily increasing die dose of prednisone. One patient who survived for four and a half months and who had a poor tissue match subsequently developed chronic rejection with progressive cholestatic jaundice. Five of the patients were able to go home and at time of publication two are alive and well 14 and 20 weeks after treatment.     

Development and differentiation of bone marrow mechanocytes

Morphological studies in the model with the complete destruction of bone marrow in the diaphysis of the tubular bone have revealed the process of stromal elements regeneration. Within the first 24 hours after the trauma insignificant accumulations of fibroblast-like cells in the lumen of some vascular channels have been observed in cell-free cortical plates. In two days spheric cells with a large dense nucleus were detected in tissue-free bone-marrow diaphysis cavity. This fact seems indicative of the viability of the vascular channel cells. This was manifested in the regained tinctorial and biological properties (i.e. the ability to proliferate). The recovery of these properties are likely to be related to vessel pericytes in the microcirculatory net of the vascular channels.     

Halothane Hepatitis—A Preventable Disease?

Eleven patients with acute hepatitis following multiple anaesthetics with halothane have been seen at two hospitals during the last four years. Six had massive hepatic necrosis and died. Unexplained fever occurred in nine of the patients; two of these and one other had had previous episodes of jaundice after halothane anaesthesia. Thus, in ten patients the disease could have been avoided.     

Diagnosis of icteric-type hepatocellular carcinoma by fine needle aspiration: a case report

BACKGROUND: Bile duct invasion is very rare in patients with hepatocellular carcinoma (HCC). It usually presents difficult problems with the clinical differential diagnosis. Moreover, another difficulty might arise when an obstructive jaundice patient is found to have past history of 2 separate malignancies. Fine needle aspiration (FNA) becomes the method of choice for clarification of the bile duct tumor thrombus. CASE: A 72-year-old man presented with progressive obstructive jaundice for 1 month. Past history revealed the occurrence of 2 distinct malignancies during the previous 3 years; they had been resected successfully. Initial imaging studies, including abdominal sonography and computed tomography, were negative for the liver. However, FNA demonstrated clusters of pleomorphic and hyperchromatic cancer cells with an increased nuclear/cytoplasmic ratio proliferating in a vague trabecular pattern with some appearance of sinusoids. Multinucleated giant cells were seen. No bile duct epithelial cells were seen. The diagnosis of the third separate malignancy, moderately differentiated HCC, was made. CONCLUSION: To our knowledge, this is the first report of icteric-type HCC diagnosed by FNA although the primary lesion was undetectable on routine, noninvasive examinations. FNA cytology is an accurate and minimally invasive method for early confirmation of biliary HCC thrombi.     

Hepatic reactions associated with ketoconazole in the United Kingdom.

Ketoconazole was introduced in the United Kingdom in 1981. By November 1984 the Committee on Safety of Medicines had received 82 reports of possible hepatotoxicity associated with the drug, including five deaths. An analysis of the 75 cases that had been adequately followed up suggested that 16, including three deaths, were probably related to treatment with the drug. Of the remainder, 48 were possibly related to treatment, five were unlikely to be so, and six were unclassifiable. The mean age of patients in the 16 probable cases was 57.9, with hepatotoxicity being more common in women. The average duration of treatment before the onset of jaundice was 61 days. None of these well validated cases occurred within the first 10 days after treatment. The results of serum liver function tests suggested hepatocellular injury in 10 (63%); the rest showed a mixed pattern. In contrast, the results of histological examination of the liver often showed evidence of cholestasis. The characteristics of the 48 patients in the possible cases were similar. Allergic manifestations such as rash and eosinophilia were rare. Hepatitis was usually reversible when treatment was stopped, with the results of liver function tests returning to normal after an average of 3.1 months. In two of the three deaths probably associated with ketoconazole treatment the drug had been continued after the onset of jaundice and other symptoms of hepatitis. Clinical and biochemical monitoring at regular intervals for evidence of hepatitis is advised during long term treatment with ketoconazole to prevent possible serious hepatic injury.     

Structure of the excretory ducts, intraorganic lymphatic vessels, tissue canals and intercellular space of various organs (data of scanning electron microscopy injection replicas)

Architectonics of the biliary canaliculi and intrahepatic ducts systems, as well as intraorganic urinary pathways in white rats have been investigated by means of scanning electron microscopy of injection replica. Acinar structure of the intralobular part of the biliary bed has been proved. Anastomoses between the biliary canaliculi of the neighbouring lobules have been demonstrated. A useful method for obtaining injection replica of the intraorganic lymphatic vessels is filling of the ductal system of the parenchymatous organs with solid resins (methylmethacrylate+) under a high (nonphysiological) pressure. Casts of periportal and paravenous hepatic lymphatic vessels have been obtained. An ability of methylmethacrylate to replicate intercellular and connective tissue spaces is verified. Casts of the perisinusoid spaces (Disse) are obtained for the first time.