Decreased Serum Alkaline Phosphatase Research Articles

Molecular and clinical analysis of ALPL in a cohort of patients with suspicion of Hypophosphatasia.

Hypophosphatasia (HPP) is a rare autosomal dominant or recessive metabolic disorder caused by mutations in the tissue nonspecific alkaline phosphatase gene (ALPL). To date, over 300 different mutations in ALPL have been identified. Disease severity is widely variable with severe forms usually manifesting during perinatal and/or infantile periods while mild forms are sometimes only diagnosed in adulthood or remain undiagnosed. Common clinical features of HPP are defects in bone and tooth mineralization along with the biochemical hallmark of decreased serum alkaline phosphatase activity. The incidence of severe HPP is approximately 1 in 300,000 in Europe and 1 in 100,000 in Canada. We present the clinical and molecular findings of 83 probands and 28 family members, referred for genetic analysis due to a clinical and biochemical suspicion of HPP. Patient referrals included those with isolated low alkaline phosphatase levels and without any additional clinical features, to those with a severe skeletal dysplasia. Thirty-six (43.3%) probands were found to have pathogenic ALPL mutations. Eleven previously unreported mutations were identified, thus adding to the ever increasing list of ALPL mutations. Seven of these eleven were inherited in an autosomal dominant manner while the remaining four were observed in the homozygous state. Thus, this study includes a large number of well-characterized patients with hypophosphatasemia which has permitted us to study the genotype:phenotype correlation. Accurate diagnosis of patients with a clinical suspicion of HPP is crucial as not only is the disease life-threatening but the patients may be offered bone targeted enzymatic replacement therapy. © 2017 Wiley Periodicals, Inc.

The ability of hesperidin compared to that of insulin for preventing osteoporosis induced by type I diabetes in young male albino rats: A histological and biochemical study

BackgroundPatients with type I diabetes are at increased risk of osteoporosis even after insulin therapy in adult stage. This study was conducted to compare the efficacy of hesperidin (hesp) therapy versus that of insulin alone in the alleviation of osteoporosis arising from type I diabetes mellitus (T1DM) in young rats. Materials and methodsHesperidin was administered orally to STZ-induced diabetes. The animals were evaluated morphologically and biochemically and compared with that received daily SC injections of long-acting insulin. ResultsHistologically, we observed the degeneration of osteoblasts and osteocytes, decreased collagen fibers, and disturbed bone turn over markers in untreated DM rats. Hesperidin+ insulin supplementation to diabetic rats caused significant improvement of most of the bone histological and morphometric parameters compared with the insulin-treated group. Furthermore, hesp treatment significantly reduced pro-inflammatory mediators TNFα and NF-κB and increased serum biochemical markers of bone turnover, including osteopontin (OPN), osteocalcin (OC) and decreased serum alkaline phosphatase (ALP). ConclusionThese data demonstrated that hesp could be considered to be a beneficial drug for preventing diabetic osteoporosis in growing age.

Effects of Simvastatin and Combination of Simvastatin and Nylestriol on Bone Metabolism in Ovariectomized Rats.

We aim to compare the effects of simvastatin and combination of simvastatin and nylestriol on bone metabolism in ovariectomized (OVX) rats. Fifty healthy Wistar female rats were randomly allocated into 5 groups: sham + saline group (group A), OVX + saline group (group B), OVX + simvastatin (5 mg·kg·d) (group C), OVX + nylestriol (0.01 mg·kg·d) (group D), and OVX + simvastatin (3 mg·kg·d) + nylestriol (0.005 mg·kg·d) (group E). All mice were orally administrated with saline or medicine dissolved in saline for 10 weeks. Body weight of rats before and after the experiment was measured. Twenty-four hours after the experiment, calcium (Ca), creatinine (Cr), and hydroxyproline in urine were detected. Serum levels of osteocalcin (bone Gla-protein, BGP) and alkaline phosphatase (ALP) were measured. Bone mineral density was detected and trabecular bone was observed after the isolation of femur and tibia. Remarkably decreased serum BGP and increased serum ALP levels were detected in group B compared with those in group A. However, notably increased serum BGP and decreased serum ALP levels were found in groups C, D, and E compared with those in group B; femoral and tibial bone mineral density decreased in group B compared with that in group A, but increased in groups C, D, and E compared with that in group B. Simvastatin and combination of simvastatin and nylestriol promote formation of new bone, increase bone density, and improve bone microstructure damage in OVX rats.

Combined Effects of Phytoestrogen Genistein and Silicon on Ovariectomy-Induced Bone Loss in Rat.

This study was performed to evaluate the effect of concomitant supplementation of genistein and silicon on bone mineral density and bone metabolism-related markers in ovariectomized rat. Three-month-old Sprague Dawley female rats were subjected to bilateral ovariectomy (OVX) or sham surgery, and then the OVX rats were randomly divided into four groups: OVX-GEN, OVX-Si, OVX-GEN-Si, and OVX. Genistein and silicon supplementation was started immediately after OVX and continued for 10weeks. In the OVX-GEN group, 5mg genistein per gram body weight was injected subcutaneously. The OVX-Si group was given soluble silicon daily in demineralized water (Si 20mg/kg body weight/day). The OVX-GEN-Si group was given subcutaneous injections of 5mg genistein per gram body weight, at the same time, given soluble silicon daily (Si 20mg/kg body weight/day). The results showed that the genistein supplementation in the OVX rats significantly prevented the loss of uterus weight; however, the silicon supplementation showed no effect on the uterus weight loss. The lumbar spine and femur bone mineral density was significantly decreased after OVX surgery; however, this decrease was inhibited by the genistein and/or silicon, and the BMD of the lumbar spine and femur was the highest in the OVX-GEN-Si-treated group. Histomorphometric analyses showed that the supplementation of genistein and/or silicon restored bone volume and trabecular thickness of femoral trabecular bone in the OVX group. Besides, the treatment with genistein and silicon for 10weeks increased the serum levels of calcium and phosphorus in the OVX rats; serum calcium and serum phosphorus in the OVX-GEN-Si group were higher than those in the OVX-GEN and OVX-Si group (P<0.05). At the same time, the treatment with genistein and/or silicon decreased serum alkaline phosphatase (ALP) and osteocalcin, which were increased by ovariectomy; serum ALP and osteocalcin in the OVX-GEN-Si group were lower than those in the OVX-GEN and OVX-Si groups (P<0.05). The results above indicate that genistein and silicon have synergistic effects on bone formation in ovariectomized rats.

Insight into the natural history of primary biliary cirrhosis: A systemic review of data from placebo-controlled clinical trials.

The natural history of primary biliary cirrhosis (PBC) is extremely variable. The extraction and analysis of available information from placebo-treated patients in randomized controlled trials of PBC treatment would facilitate the study of the natural history of PBC. The aim of the present study was to determine important clinical information regarding the natural history of PBC patients without effective treatment. A search of the PubMed, EMBASE and Cochrane Library databases was performed by two authors. Twelve randomized placebo-controlled clinical trials for PBC patients without decompensated cirrhosis were retrieved for further review. Pooled estimates of biochemical measurements, histological scores and clinical outcomes associated with PBC were calculated in the placebo group. Placebo-treated PBC patients displayed a significant decrease in serum alkaline phosphatase and very slight fluctuations in the other biochemical parameters during the 2-year follow-up. Meanwhile, histological progression was observed in 39.4% of the placebo-treated patients, and a moderate deterioration in histological scores was noted after 2 years. The pooled 2-year rates of death, transplantation and development of varices were 11.4%, 8.7% and 10.6%, respectively, in placebo-treated PBC patients. This review provides a foundation for further epidemiologic investigations in untreated patients and ursodeoxycholic acid-resistant patients with PBC. Biochemical responses after 2 years may provide some information on disease progression and therapeutic response in PBC.

Coconut water drink and the risk of hyperkalaemia in diabetes

AbstractCoconut water is available as a substitute for table water and as a sports rehydrating drink. Because of the nature of its micronutrients, it may lead to biochemical changes that may not be beneficial for all groups of people.A patient with type 2 diabetes mellitus consumed daily around one litre of coconut water drink. As a result, there was a gradual increase in serum potassium. On cessation of beverage consumption, serum potassium decreased to within the reference interval. However, an increase in urea and creatinine concentration did not revert to the level seen prior to coconut water consumption. There was a decrease in serum alkaline phosphatase and zinc when consuming the beverage. Reduction occurred in diastolic blood pressure, estimated glomerular filtration rate, serum enzymes and zinc, while serum potassium concentration increased in this patient with type 2 diabetes. The observed changes resulted from consuming excessive quantities of coconut water drinks.In patients with diabetes and renal impairment and on potassium‐retaining medication, there is a high risk of developing hyperkalaemia. Copyright © 2016 John Wiley &amp; Sons.

Mutagenicity and preclinical safety assessment of the aqueous extract of Clinacanthus nutans leaves

Context: Clinacanthus nutans (CN) is used traditionally for treating various illnesses. Robust safety data to support its use is lacking. Objective: To evaluate the adverse effects of aqueous extract of CN leaves (AECNL). Materials and methods: The oral toxicity of the AECNL was tested following Organisation for Economic Co-operation and Development (OECD) guidelines. Mutagenicity (Ames test) of AECNL was evaluated using TA98 and TA100 Salmonella typhimurium strains. Results: No mortality or morbidity was found in the animals upon single and repeated dose administration. However, significant body weight loss was observed at 2000 mg/kg during sub-chronic (90 d) exposure. In addition, increased eosinophil at 500 mg/kg and decreased serum alkaline phosphatase levels at 2000 mg/kg were observed in male rats. Variations in glucose and lipid profiles in treated groups were also observed compared to control. Ames test revealed no evidence of mutagenic or carcinogenic effects at 500 μg/well of AECNL. Conclusion: The median lethal dose (LD50) of the AECNL is >5000 mg/kg and the no-observed-adverse-effect level is identified to be greater than 2000 mg/kg/day in 90-d study.

Effect of dietary cottonseed meal on growth performance, physiological response, and gossypol accumulation in pre-adult grass carp, Ctenopharyngodon idellus

Cottonseed meal (CM) was used at up to 36.95% content in the diet (replacing 60% of dietary fish meal protein) without any negative eff ects on growth performance of pre-adult grass carp (initial body weight, 761 g) under outdoor conditions. A culture trial was conducted in net cages installed in a large concrete pond. Seven isonitrogenous and isoenergetic diets containing a gradient of CM concentrations (0, 12.2%, 24.4%, 36.6%, 48.8%, 54.8%, and 61.0%) as replacement for dietary fish meal protein (0, 20%, 40%, 60%, 80%, 90%, and 100%) were formulated. Dietary non-resistant starch (from maize) was inverse to dietary CM. Growth performance and feed utilization of fish fed the diets containing CM replacing 0–40% fishmeal protein were not aff ected after the 6-week feeding trial. Accumulation of hepatopancreatic total gossypol in the hepatopancreas was significantly correlated with free gossypol content in the diets (HTG=88.6+1.5×DFG, R2=0.89, P<0.05). Intestinal α-amylase and γ-glutamyl transpeptidase activities rose along with increasing dietary CM level. The structure of the mid-intestinal tissues and the ultrastructure of the enterocyte microvilli were normal when dietary CM was <36.6% (60% protein replacement). Increasing dietary CM content increased serum alanine aminotransferase levels but decreased serum alkaline phosphatase, cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triglycerides, and albumin (P<0.05).

Effects of Radiation Therapy on Established Neurogenic Heterotopic Ossification

Heterotopic ossification (HO) is frequently seen on rehabilitation units after spinal cord injuries, fractures, brain injuries, and limb amputations. Currently, there is no effective treatment for HO other than prophylaxis with anti-inflammatory medications, irradiation, and bisphosphonate administration. These prophylactic treatments are not effective for managing ectopic bone once it has formed. Here we describe three cases of established neurogenic HO treated with radiation therapy (RT). All patients had decreased serum alkaline phosphatase (ALP) and bone-specific ALP levels with decreased pain but increased range of motion immediately after RT. Post-treatment X-rays revealed no further growth of the HO. All patients maintained clinical and laboratory improvements 4 or 6 months after the RT. Our results suggest that RT is safe and effective in decreasing pain and activity of neurogenic HO.

Bone Density, Osteocalcin and Deoxypyridinoline for Early Detection of Osteoporosis in Obese Children.

AIM:This study aimed at comparing between bone density using DEXA, serum osteocalcin and urinary DPD in obese and non obese prepubertal children.METHODS:After taking the consent of eighty children they were subjected to: full examination, anthropometric measurements, blood samples were withdrawn to determine serum osteocalcin, Ca, Ph, alkaline phosphatase, and urinary DPD. Bone densities, body composition of the whole body were examined using DEXA. Data were analyzed using SPSS.RESULTS:All anthropometric variables showed significant increase in obese children except for height in comparison to control group. Total mass, lean + BMC, lean, fat, area, BMC, BMD and Z score of the whole body were significantly increased in obese children. Serum calcium showed significant increase while alkaline phosphatase was significantly decreased in obese children. DPD showed no significant difference between obese and non obese children. Significant negative correlation was found between ca, lean, lean + BMC and total mass. Serum alkaline phosphatase showed also a significant negative correlation with (lean + BMC and total mass). Serum osteocalcin showed negative significant correlation with area, BMC, BMD, lean and Z score.CONCLUSION:Obese children showed significant increase in anthropometric and DEXA parameters, increase in serum calcium and significant decrease in serum alkaline phosphatase. Osteocalcin was negatively correlated with most of DEXA results.

Dynamic changes in biomarkers for immune response to sipuleucel-t (SipT) for metastatic castrate-resistant prostate cancer (mCRPC).

181 Background: SipT prolongs OS in asymptomatic or minimally symptomatic CRPC without regularly inducing PSA declines. Elevated CD54 count, CD54 upregulation (UPREG) and total nucleated cells (TNC), markers of immune activation in the product, have been shown to correlate with longer survival. We prospectively collected clinical characteristics and inflammatory markers as part of an audit to identify possible predictors of immune response in patients receiving SipT. Methods: 92 men with CRPC had blood drawn routinely before and after SipT treatment at USC. Circulating tumor cells (CTC), PSA, prostatic acid phosphatase (PAP), albumin (Alb), hemoglobin (Hb), serum alkaline phosphatase (SAP), LDH, C-reactive protein (CRP) and β2-microglobulin (β2m) were evaluated in a reference lab. SipT product parameters, CD54, UPREG and TNC were measured by Dendreon. These datasets were analyzed using Spearman coefficients for continuous variables and the Kruskal-Wallis test for categorical variables. Results: 92 pts who received SipT were included. Median age was 69 (48-90). 43% had Gleason 8-9. 42% had ECOG Performance Status (PS) 0, 44% PS 1, and 13% PS 2. 26% had received prior chemotherapy and 18% had used opioids. 84% had bone mets, 42% had lymphadenopathy, and 12% had visceral mets. Increases in the PSA, PAP, LDH and Alb was directly correlated with increased CD54 counts, increases in the CTC was directly correlated with increased TNC, whereas decrease in SAP was correlated with increased upregulation of dendritic cells. Conclusions: Our results show that increased tumor markers immediately after treatment are more common in high CD54 product pts. This may represent a “flare” due to immune activation. [Table: see text]

Benign prenatal hypophosphatasia: a treatable disease not to be missed

Prenatal bowing of the long bones is often associated with severe bone dysplasias. We report a child who presented marked bowing of the long bones at birth but showed a relatively benign postnatal course with spontaneous improvement of bowing. The fetal imaging showed normal skeletal mineralization and normal chest and abdominal circumferences despite the limb bowing and shortening. Decreased serum alkaline phosphatase activity and elevated urine phosphoethanolamine was biochemically evident, and compound heterozygous mutations in the tissue-nonspecific alkaline phosphatase (TNSALP) gene were identified, which confirmed the diagnosis of a benign form of prenatal hypophosphatasia. Benign prenatal hypophosphatasia should be considered in the differential diagnosis of congenital bowing of the long bones.

Impact of caffeic acid phenethyl ester treatment on vancomycin-induced pancreatic damage in rats

This study investigates the preventive effect of caffeic acid phenethyl ester (CAPE) on pancreatic damage induced by vancomycin (VCM) in rats. Rats were equally divided into three groups: group I (control), group II (only VCM-treated group) and group III (VCM + CAPE-treated groups). VCM was intraperitoneally administered at a dose of 200 mg kg(-1)twice daily for 7 days. CAPE was administered orally at 10 µM mL(-1) kg(-1) dose once daily for 7 days. The first dose of CAPE administration was performed 24 h prior to VCM injection. Blood and pancreas tissue samples were removed and collected after the study. Serum alkaline phosphatase (ALP), amylase, γ-glutamyl transferase (GGT) and lipase activities were determined. Pancreas tissue samples were evaluated with the light microscope. Group II significantly increased serum ALP, amylase, GGT and lipase activities when compared with the control group. Group III significantly decreased serum ALP, amylase, GGT and lipase activities when compared with group II. In histopathological examination, it has been observed that there was a significant pancreatic damage in group II. CAPE exerted prominent structural protection against VCM-induced pancreatic damage and this effect was statistically significant. CAPE caused a marked reduction in the extent of pancreatic damage. We have concluded that it may play an important role in the VCM-induced pancreatic damage and reduce the pancreatic damage both at the biochemical and histopathological aspects.

A pilot study of umbilical cord‐derived mesenchymal stem cell transfusion in patients with primary biliary cirrhosis

Ursodeoxycholic acid (UDCA) treatment is an effective medical therapy for patients with primary biliary cirrhosis (PBC); however, 40% of PBC patients show an incomplete response to the UDCA therapy. This study aimed to investigate the safety and efficacy of umbilical cord-derived mesenchymal stem cell (UC-MSC) transfusion in PBC patients with an incomplete response to UDCA. We conducted a single-arm trial that included seven PBC patients with a suboptimal response to UDCA treatment. UC-MSCs were first cultured, and then 0.5 × 10(6) cells/kg body weights were infused through a peripheral vein. UC-MSCs were given three times at 4-week intervals, and patients were followed up for 48 weeks. Primary outcomes were to evaluate the safety and feasibility of UC-MSC treatment, and secondary outcomes were to evaluate liver functions and patient's quality of life. No obvious side-effects were found in the patients treated with UC-MSCs. Symptoms such as fatigue and pruritus were obviously alleviated in most patients after UC-MSC treatment. There was a significant decrease in serum alkaline phosphatase and γ-glutamyltransferase levels at the end of the follow-up period as compared with baseline. No significant changes were observed in serum alanine aminotransferase, aspartate aminotransferase, total bilirubin, albumin, prothrombin time activity, international normalized ratio, or immunoglobulin M levels. The Mayo risk score, a prognostic index, was also stable during the treatment and follow-up period. UC-MSC transfusion is feasible and well tolerated in patients with PBC who respond only partially to UDCA treatment, thus representing a novel therapeutic approach for patients in this subgroup. A larger, randomized controlled cohort study is warranted to confirm the clinical efficacy of UC-MSC transfusion.

Extraction and bioactivity of polygonatum polysaccharides

The present study is to explore the optimal extraction parameters and liver protective effect of the polygonatum polysaccharides in vivo. The order of factor effects on polysaccharides production was found to be extraction time (min, A)>ratio of solvent to solid (C)>extraction temperature (°C, B)>extraction number (D). The results show that the effects of extraction time (min, A) and ratio of solvent to solid (C) were more significant than those of the other factors. Optimal extraction parameters were as followings: extraction time 120min, extraction temperature 100°C, ratio of solvent to solid 5, and extraction number 4. Polygonatum polysaccharides was administered orally at doses of 150, 300 and 450mg/(kg day) to carbon tetrachloride (CCl4)-treated rats. Results showed that administration of polygonatum polysaccharides could increase rats’ final body weight, liver antioxidant enzymes activities (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and glutathione reductase (GR)), decrease serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) activities and liver malondialdehyde (MDA) level. The liver sections obtained from animals supplemented with polygonatum polysaccharides extract demonstrated reduced pathological damages, supporting that polygonatum polysaccharides extract could effectively decrease the toxicity of CCl4. It can be concluded that polygonatum polysaccharides treatment may prevent CCl4-induced liver oxidative injury in experimental rats.

Effects of feed-borne Fusarium mycotoxins on performance, serum chemistry, and intestinal histology of New Zealand White fryer rabbits1

The objective of the present study was to determine the effects of feeding diets containing grains naturally contaminated with Fusarium mycotoxins to fryer rabbits. The efficacy of a glucomannan mycotoxin adsorbent (GMA) was also examined. Thirty 5-wk-old male New Zealand White rabbits were fed a control diet, a contaminated diet, or a contaminated diet + 0.2% GMA for 21 d. Experimental diets contained deoxynivalenol (DON; vomitoxin) at a concentration of 0.25, 4.3, and 4.9 μg/g, respectively. Feed intake was measured daily and water intake was measured every 3 d. At the end of the feeding period, blood was collected for evaluation of serum chemistry and hematology. Visceral organs were excised, weighed, and processed for histopathological examination. Body weight gain and water intake were greater in rabbits fed the contaminated diet (P = 0.075 and 0.020, respectively) and those fed the contaminated + GMA diet (P = 0.026 and 0.002, respectively) compared with controls. Rabbits fed the contaminated + GMA diet had significantly increased serum urea concentrations (P = 0.023) and decreased serum alkaline phosphatase activity (P = 0.020) compared with controls. Increase in BW gain of rabbits fed the contaminated diets was caused by increased water consumption. There was no effect (P > 0.05) of diet on relative organ weights, but decreased infiltrations with eosinophilic granulocytes were observed in different regions of the intestine in rabbits fed the contaminated or the contaminated + GMA diet. It was concluded that rabbits could be adversely affected by feed-borne Fusarium mycotoxins but appear to be less sensitive than other mammalian species. Supplementation with GMA did not reduce many of the effects of feed-borne mycotoxins.

Protective effect of Morinda officinalis polysaccharides on bone degeneration in theaged rats

In the current study, we examined the effect of Morinda officinalis polysaccharides (MOP) on bone quality in aged rats. Rats were given orally by daily gavage at 200 and 400 mg/kg/day for 3 months, respectively. The results showed that there was a significant decrease in serum alkaline phosphatase (ALP), Interleukin 6 (IL-6) content and a significant increase in density and strength of femur and lumbar vertebrae in aged rats 3 months afterMorinda officinalis polysaccharides treatment. In conclusion, these data suggest thatMorinda officinalis polysaccharides can protect the age-induced bone degeneration. Key words: Morinda officinalis polysaccharides, bone, osteoporosis, ALP.

Osteoprotective effect of Phaseolus vulgaris L in ovariectomy-induced osteopenia in rats

Phytoestrogens and phytoestrogen-containing plants are currently being explored as potential candidates for the treatment of estrogen-related disorders. The aim of this study was to evaluate the anti-osteoporotic effect of the phytoestrogen-rich plant Phaseolus vulgaris L, commonly known as French beans. Adult Sprague-Dawley rats were either bilaterally ovariectomized (OVX) or sham operated. OVX and sham control groups were administered vehicle, whereas the other two OVX groups were given 0.15 mg/kg estradiol and 1 g/kg methanolic extract of P vulgaris L seeds (MPV) orally for 10 weeks (10 rats per group). At autopsy, blood, urine, bones, and uteri of the animals were collected. Serum was evaluated for estradiol, calcium (Ca), phosphorus, alkaline phosphatase, tartarate resistant acid phosphatase, and urine for Ca. The bone density, ash density, mineral content, and mechanical strength of bones was evaluated. Scanning electron microscopic analysis of bones (tibia) was performed. Results were analyzed using analysis of variance and Tukey's multiple comparison test. Compared with the OVX control, MPV (1 g/kg PO) significantly decreased serum alkaline phosphatase and reduced serum tartarate resistant acid phosphatase and urinary Ca levels. It caused an increase in bone density, ash density, and bone mechanical strength and significantly increased bone Ca. Improvement in bone microarchitecture was indicated by image analysis of scanning electron microscopic photomicrographs. No increase in weight of atrophic uterus in OVX animals was observed with MPV treatment. Treatment with MPV prevented estrogen deficiency-induced osteopenia without affecting the uterine mass. The promising results of the study warrant further investigation of French beans as a potential candidate for the treatment of postmenopausal osteoporosis.

Ursodeoxycholic acid for treatment of cholestasis in patients with hepatic amyloidosis

Amyloidosis represents a group of different diseases characterized by extracellular accumulation of pathologic fibrillar proteins in various tissues and organs. Severe amyloid deposition in the liver parenchyma has extrahepatic involvement predominantly in the kidney or heart. We evaluated the effect of ursodeoxycholic acid, in four patients with severe hepatic amyloidosis of different etiologies, who presented with increased alkaline phosphatase and gamma-glutamyl transferase. The study included four patients who presented with amyloidosis-associated intrahepatic cholestasis. Three of them had renal amyloidosis which developed 1-3 years before cholestasis occurred, the remaining one having intrahepatic cholestasis as the primary sign of the disease. Amyloidosis was identified from liver biopsies in all patients by its specific binding to Congo red and green birefringence in polarized light. The biochemical nature and the class of amyloid deposits were identified immunohistochemically. In addition to their regular treatment, the patients received 750 mg ursodeoxycholic acid per day. After 2-4 weeks all patients had a significant decrease of serum alkaline phosphatase and gamma-glutamyl transferase, and their general status significantly improved. Treatment with ursodeoxycholic acid may be beneficial in patients with hepatic amyloidosis, and do extend indications for the use of ursodeoxycholic acid in amyloidotic cholestatic liver disease.

Role of hepatic phospholipids in development of liver injury in Mdr2 (Abcb4) knockout mice

Multidrug resistance protein 2 (Abcb4) gene knockout mice (Mdr2(-/-)) lack phosphatidylcholine (PC) excretion into bile and spontaneously develop sclerosing cholangitis, biliary fibrosis and hepatocellular carcinomas. We therefore aimed to test whether formation and hepatic retention of abnormal PC metabolites contribute to the pathogenesis of liver injury in Mdr2(-/-) mice. Mdr2(-/-) mice were either fed a diet supplemented with soybean lecithin 2.5% w/w [phosphatidylcholine-enriched diet (PCD), to increase hepatic PC content] or a choline-deficient diet (CDD, to reduce hepatic PC content) for 4 weeks; controls received chow with energy and nutrient content equivalent to PCD and CDD. Serum liver tests, liver histology, markers of fibrosis, cholangiocyte activation, cell proliferation and thin-layer chromatography for phospholipid (PL) composition were carried out. PCD decreased serum alkaline phosphatase and total bilirubin levels compared with controls, while liver histology as well as hepatic hydroxyproline content as markers of liver fibrosis did not differ among groups. Both PCD and CDD decreased hepatocellular proliferation compared with controls. Hepatic, serum and biliary PLs remained unchanged despite dietary manipulations and no potentially toxic PL metabolites were detected. Mdr2(-/-) mice maintain stable hepatic, serum and biliary PL metabolism in response to dietary PC manipulations. Our findings therefore suggest that liver injury in Mdr2(-/-) mice is not due to formation of toxic PL metabolites.