Decreased Serum Alkaline Phosphatase Research Articles

The Thyromimetic Sobetirome (GC-1) Alters Bile Acid Metabolism in a Mouse Model of Hepatic Cholestasis

Chronic cholestasis results from bile secretory defects or impaired bile flow with few effective medical therapies available. Thyroid hormone triiodothyronine and synthetic thyroid hormone receptor agonists, such as sobetirome (GC-1), are known to impact lipid and bile acid (BA) metabolism and induce hepatocyte proliferation downstream of Wnt/β-catenin signaling after surgical resection; however, these drugs have yet to be studied as potential therapeutics for cholestatic liver disease. Herein, GC-1 was administered to ATP binding cassette subfamily B member 4 (Abcb4-/-; Mdr2-/-) knockout (KO) mice, a sclerosing cholangitis model. KO mice fed GC-1 diet for 2 and 4 weeks had decreased serum alkaline phosphatase but increased serum transaminases compared with KO alone. KO mice on GC-1 also had higher levels of total liver BA due to alterations in expression of BA detoxification, transport, and synthesis genes, with the net result being retention of BA in the hepatocytes. Interestingly, GC-1 does not induce hepatocyte proliferation or Wnt/β-catenin signaling in KO mice, likely a result of decreased thyroid hormone receptor β expression without Mdr2. Therefore, although GC-1 treatment induces a mild protection against biliary injury in the early stages of treatment, it comes at the expense of hepatocyte injury and is suboptimal because of lower expression of thyroid hormone receptor β. Thus, thyromimetics may have limited therapeutic benefits in treating cholestatic liver disease.

Functional bioactive substance improves the growth performance, antioxidant capacity and immune function of growth retardation pigs

ABSTRACT This study was conducted to investigate the effects of composited functional bioactive substances on the growth performance and immune function of growth retardation (GR) pigs. Twenty-four 42-day-old GR pigs were fed with basal diet containing 0 or 0.5% composited functional bioactive substances package (FP) for 21 days. The results showed that dietary supplementation with FP decreased feed:gain ratio, increased the relative and absolute weight of the liver (P < 0.05). Supplementation with FP decreased serum alkaline phosphatase and malondialdehyde, but increased aspartic acid, cysteine, arginine, proline and taurine concentrations, as well as serum superoxide dismutase and catalase activities (p < 0.05). FP treatment increased all determined cytokines (IL-1β, IL-4, IL-6, IL-8, IL-10, IL-12, GM-CSF, IFN-γ, TGF-β1, TNF-α) concentrations in serum compared with the control treatment (p < 0.05). These findings indicate that functional bioactive substances can improve feed conversion rate, serum anti-oxidant capacity and immunity and can be as effective growth promoter in the GR piglets.

Growth performance, nutrient digestibility, bone mineralization, and hormone profile in broilers fed with phosphorus-deficient diets supplemented with butyric acid and Saccharomyces boulardii

The present study evaluated the effects of butyric acid supplementation and Saccharomyces boulardii (alone or in combination) on growth performance, nutrient digestibility, bone mineralization, and blood hormones of male broiler chickens fed a diet including reduced levels of nonphytate phosphorus (NPP). The chickens were allocated to 6 dietary treatments: 1) positive control diet with adequate amounts of NPP (PC; 0.48, 0.43, and 0.39% in the starter, grower, and finisher period, respectively); 2) negative control diet with low amounts of NPP (NC; 0.38, 0.33, and 0.29% in the starter, grower, and finisher period, respectively); 3) NC plus 500 FTU/kg microbial phytase (PHY); 4) NC plus 0.2% butyric acid (BA); 5) NC plus 1 × 108 cfu/kg S. boulardii (SB); 6) NC plus butyric acid and S. boulardii (BA+SB). Each treatment had 5 pen replicates of 25 birds. After 6 wk, the body weight and ADG in birds fed with any of the diets were higher (P < 0.001) than those in birds fed with the NC diet, where the birds fed with the PHY and BA+SB diets had the highest values. However, only the PHY diet improved (P = 0.041) overall F:G. All diets, except the SB diet, resulted in the increased apparent ileal digestibility coefficient (AIDC) of CP, AMEn, and tibia ash content and decreased serum alkaline phosphatase level compared with the NC diet (P < 0.05). Broiler chickens fed with the PHY, SB, and BA+SB diets also had increased AIDC of phosphorus (P = 0.017) than those fed with the NC and PC diets. Feeding PC, PHY, and BA+SB diets increased (P = 0.007) the tibia phosphorus content but decreased (P = 0.033) serum parathyroid hormone concentration. Overall, the present data indicate that the simultaneous inclusion of butyric acid plus S. boulardii in the low-NPP diets was beneficial for improving growth rate and bone mineralization, but not for feed efficiency.

1067 Use of Obeticholic Acid in Patients With Primary Biliary Cholangitis: A Systematic Review and Meta-Analysis

INTRODUCTION: Obeticholic acid (OCA), a farnesoid-X receptor agonist, has been approved for the management of PBC since 2016. It has been shown to significantly decrease serum alkaline phosphatase (ALP) and total bilirubin levels in patients with primary biliary cholangitis (PBC), either alone or in combination with ursodiol. Data on the efficacy of OCA in PBC is limited. We conducted a systematic review and meta-analysis to evaluate the efficacy of OCA in patients with PBC. METHODS: We searched PubMed, Cochrane and Embase databases for randomized control trials (RCTs) comparing OCA to placebo in the management of PBC through 2019. The Cochrane Risk of Bias tool was used to assess the quality of the RCTs. We estimated odds ratio (OR) with 95% confidence interval (CI) for each outcome using a random effects model. The quality of the evidence was assessed using the GRADE approach. RESULTS: A total of 3 randomized controlled trials including 440 patients were identified. The mean age of patients was 54 and follow up ranged from 3-12 months. The studies varied with respect to whether OCA was used as monotherapy or added to standard of care (ursodiol). Compared to placebo, OCA resulted in reduction of ALP to &lt; 1.67X the ULN with an OR of 5.92 (CI 2.98 – 11.94, P &lt; 0.00001); and 1.76X the ULN with an OR of 5.95 (CI 3.25 – 10.93; P &lt; 0.00001). Across the three studies, patients on OCA experienced higher rates of pruritus as compared with placebo. The overall certainty of evidence was moderate. Data on clinical endpoints such as quality of life, progression to cirrhosis or liver transplantation, and mortality were lacking. CONCLUSION: Obeticholic acid is superior to placebo in reducing ALP in patients with PBC. An important limitation in these studies is the lack of data on clinical endpoints however, ALP levels have been shown to correlate with risk of liver transplantation and death. More RCT data with longer follow up is needed to fully assess the net benefits and harms of OCA in patients with PBC.

Smart probe for simultaneous detection of copper ion, pyrophosphate, and alkaline phosphatase in vitro and in clinical samples

Wilson’s disease (WD), which might lead to acute liver failure, is an inherited disorder characterized by accumulation of copper (Cu2+) in the brain, the liver, and other vital organs. In the clinic, decreased serum alkaline phosphatase (ALP) concentration is used for WD diagnosis. But to the best of our knowledge, using a fluorescent probe to simultaneously detect multiple factors in WD (e.g., Cu2+, pyrophosphate (PPi), and ALP) has not been reported. Herein, we rationally designed a fluorescent switch (E)-8-((4-methylbenzylidene)amino)napthalen-1-amine (L) and successfully applied it for sequential and selective detections of Cu2+, PPi, and ALP in vitro, in living cells and synovial fluid samples with “Off,” “On,” and “Off” fluorescence signals, respectively. Considering the obvious correlations among Cu2+, PPi, and ALP in WD, we envision that our fluorescent probe L could be applied to in vitro diagnosing WD in the near future.

Sub-acute toxicity profile of methanolic leaf extract of Securidaca longipedunculata in rats

This study investigates the toxicological profile of Securidaca longipedunculatain rats. Sub- acute toxicity study was conducted by oral administration of the extracts to rats at daily doses of 25, 50 and 100mg/kg body weight for 28 days. Methanol leaf extract of S. longipedunculata caused significant (p&lt; 0.05) increase in serum urea, creatinine, sodium &amp; proteins but significantly (p&lt; 0.05) decreased the serum alkaline phosphatase (ALP) and cholesterol concentration when compared with the controls. However, there was no significant (p&gt; 0.05) difference in serum aspartate transaminase (AST), alanine transaminase (ALT), potassium, triglyceride albumin and bilirubin concentration when compared with control. Jobelyn and dexamethazone caused significant (p&lt; 0.05) increase in serum AST, urea, triglycerides, sodium and protein. Dexamethazone caused significant increase (p&lt;0.05) in bilirubin, cholesterol, and a decrease in serum ALP and creatinine concentration when compared with the control. The extract also had no significant (p&gt;0.05) effect on hematological parameters except for a significant increase (p&lt;0.05) in white blood cells when compared with the normal control. The plant extracts have shown no serious adverse effect on hematological and biochemical parameters. Thus, S. longipedunculata methanol leaf extract could be considered as a natural source of antibiotics for therapeutic purposes.&#x0D;

Effect of Chlorella Pyrenoidosa Protein Hydrolysate-Calcium Chelate on Calcium Absorption Metabolism and Gut Microbiota Composition in Low-Calcium Diet-Fed Rats.

In our current investigation, we evaluated the effect of Chlorella pyrenoidosa protein hydrolysate (CPPH) and Chlorella pyrenoidosa protein hydrolysate-calcium chelate (CPPH-Ca) on calcium absorption and gut microbiota composition, as well as their in vivo regulatory mechanism in SD rats fed low-calcium diets. Potent major compounds in CPPH were characterized by HPLC-MS/MS, and the calcium-binding mechanism was investigated through ultraviolet and infrared spectroscopy. Using high-throughput next-generation 16S rRNA gene sequencing, we analyzed the composition of gut microbiota in rats. Our study showed that HCPPH-Ca increased the levels of body weight gain, serum Ca, bone activity, bone mineral density (BMD) and bone mineral content (BMC), while decreased serum alkaline phosphatase (ALP) and inhibited the morphological changes of bone. HCPPH-Ca up-regulated the gene expressions of transient receptor potential cation V5 (TRPV5), TRPV6, calcium-binding protein-D9k (CaBP-D9k) and a calcium pump (plasma membrane Ca-ATPase, PMCA1b). It also improved the abundances of Firmicutes and Lactobacillus. Bifidobacterium and Sutterella were both positively correlated with calcium absorption. Collectively, these findings illustrate the potential of HCPPH-Ca as an effective calcium supplement.

In vivo anti-arthritic effect and repeated dose toxicity of standardized methanolic extracts of Buddleja cordata Kunth (Scrophulariaceae) wild plant leaves and cell culture

Ethnopharmacological relevanceBuddleja cordata Humb. Bonpl. & Kunth, known by the population as Tepozán blanco, is a shrub plant used in traditional herbal medicine in Mexico for the treatment of tumors, cancer, sores, skin burns, rheumatic pains and diseases related to inflammatory processes such as arthritis; authors adjudicate this etno-medicinal effect to the presence of secondary metabolites in the plant such as verbascoside, however due to its low concentration in recent years biotechnological tools are applied as cell culture to biosynthesize these pharmacological active metabolites in greater quantities.Aim of the study: Evaluate the possible toxic effect after a daily administration of MeOH extracts from wild plant leaves (Bc-Wp), and cell culture (Bc-Cc) of B. cordata for 28 days, and after their anti-edematous and antioxidant activities in vivo, as well their effect on the cytokines profile during experimental arthritis induced by complete Freund's adjuvant. Materials and methodsBoth extracts were evaluated in CD1 male mice first in a toxicity test of repeated dose administrations (1 g/kg) for 28 days, after which pharmacological activity of both extracts was measure during experimental induced arthritis where three doses were tested, at the end of the study edema formation, body weight gain and antioxidant activity were measure in edema and ganglionic tissues. Finally, dose that exerted the best protective effect (250 mg/kg) was evaluated to quantify its effect over ganglionic tissue concentration of lymphocytes T CD4+, and cytokines (IL-1β, TNF-α and IL-10), as well histological analysis in arthritic mice. ResultsBoth extracts (Bc-Wp and Bc-Cc) did not exert lethality, however body weight gain and food in-take were lower than in healthy mice administered with vehicles, also extract-treated animals showed a decrease in serum lipid concentration and only Bc-Wp extract treated animals decrease serum alkaline phosphatase after 28 daily administration compared to healthy un-treated mice. During experimental arthritis best inhibition effect over edema development was observed in those animals administered with both extracts at dose of 250 mg/kg (Bc-Wp and Bc-Cc) on day 28, compared to CFA un-treated mice. Also both extracts reduce oxidative damage over lipids and proteins at the same dose, in both ganglionic and edema tissue, as well antioxidant enzymatic response was reduced in both tissues compared to arthritic un-treated group. In ganglionic tissue of arthritic mice, CD4+ lymphocytes concentration was reduced by Bc-Wp and Bc-Cc treatment (250 mg/kg) respectively, as well IL-1β, and TNF-α levels. Only arthritic animals treated with Bc-Cc extract at 250 mg/kg generated a significant increase of IL-10 doubling the levels compared to CFA un-treated group. Histological analysis of popliteal ganglion showed that both extracts decrease the incidence of lytic lesions, lipid inclusions and leukocyte infiltration. ConclusionMethanolic extracts of wild Buddleja cordata and its cell cultures did not generated lethality after a daily administration for 28 days at 1 g/kg, but it was observed that both showed a lipid-lowering effect. Also at dose of 250 mg/kg both extracts exerted anti-edematous, protection against the oxidation of lipid and proteins, regulation on antioxidant enzymatic response, down-regulation on lymphocytes CD4+ producers of IL-1β and TNF-α, an increase in IL-10 levels, which caused a decrease in leukocyte infiltration in ganglionic tissue during experimental arthritis.

Turmeric and chicory seed have beneficial effects onobesity markers and lipid profile in non-alcoholic fatty liver disease (NAFLD).

In an attempt to investigate new strategies aimed at reducing risk factors of non-alcoholic fatty liver disease (NAFLD), effects of turmeric (Curcuma longa L.) and chicory seed (Cichorium intybus L.) supplementation was evaluated in these patients. In this double-blind, randomized, controlled clinical trial, 92 patients with NAFLD aged 20-60year with body mass index (BMI) ranged 24.9-40kg/m2 was randomly assigned to 4 groups as follows. 1) Turmeric supplementation (3g/d) (n=23, TUR); 2) Chicory seed supplementation (infused 9g/d (4.5g /100mL)) (n=23, CHI); 3) Turmeric and chicory seed supplementation (3g/d turmeric+infused 9g/d chicory seed (n=23, TUR+CHI); 4) Placebo (n=23, PLA). All intervention periods were 12 weeks. Fasting blood samples, anthropometric measurements, dietary records and physical activity were collected at baseline and at the end of the trial. Significant decreases were observed in BMI and waist circumference (WC) of subjects in CHI and TUR+CHI groups, compared with PLA group (p<0.05). Combination of turmeric and chicory seed significantly decreased serum alkaline phosphatase level (p<0.05). Serum levels of HDL-C increased considerably in TUR and TUR+CHI groups (p<0.05 vs. placebo). Turmeric supplementation alone and plus chicory seed led to significant reduction in serum levels of TG/HDL-C and LDL-C/HDL-C ratio in TUR and TUR+CHI groups in comparison with placebo (p<0.05). In conclusion, turmeric and chicory seed supplementation can be significantly useful in management of NAFLD risk factors.

Obeticholic acid in primary biliary cholangitis: where we stand.

This review will summarize the use of obeticholic acid (OCA) in treatment of primary biliary cholangitis (PBC). It seeks to discuss the mechanism of action, evidence for use, appropriate clinical use, and common adverse effects of OCA. PBC is a chronic, progressive cholestatic liver disease that is a chronic progressive that may lead to end-stage liver disease and need for liver transplantation. Ursodeoxycholic acid (UDCA) has been the mainstay of therapy for PBC for decades. Recent research has led to the discovery that bile acids act as hormones and have many effects, one of which is activating the farnesoid X receptor (FXR). Activation of FXR leads to decreased bile acid synthesis, inflammation, and fibrosis of the liver. OCA is a highly potent FXR agonist. Several clinical trials demonstrated that OCA treatment in PBC led to a significant decrease in serum alkaline phosphatase, a marker for long-term survival. The US FDA-approved OCA in 2016, which led to incorporation of OCA into current guidelines as a second-line treatment for PBC. The most clinically relevant adverse effect of OCA is dose-related pruritus. We review the role of OCA and current guidelines in treatment of PBC.

Denosumab improves clinical manifestations of hypophosphatemic osteomalacia by adefovir-induced Fanconi syndrome: a case report

BackgroundAdefovir dipivoxil is a nucleotide analogue that is approved for treatment of chronic hepatitis B. Adefovir dipivoxil is associated with proximal tubular dysfunction, resulting in Fanconi syndrome, which can cause secondary hypophosphatemic osteomalacia. We describe a case of a patient with hypophosphatemic osteomalacia secondary to Fanconi syndrome induced by adefovir dipivoxil concomitantly with osteoporosis in whom clinical symptoms were improved by adding denosumab (a human monoclonal antibody targeting the receptor activator of nuclear factor-κB ligand) to preceding administration of vitamin D3.Case presentationA 60-year-old Japanese man had been receiving low-dose adefovir dipivoxil (10 mg/day) to treat chronic hepatitis B for approximately 5 years. He presented to an orthopedic surgeon with severe pain of the right hip and no trauma history, and fracture of the neck of the right femur was identified. In addition, 99mTc-hydroxymethylene diphosphate scintigraphy revealed significantly abnormal uptake in the bilateral ribs, hips, and knees, and he was therefore referred to our university hospital for evaluation of multiple pathological fractures. We diagnosed hypophosphatemic osteomalacia due to Fanconi syndrome induced by adefovir dipivoxil therapy. Although we reduced the patient’s adefovir dipivoxil dose and added calcitriol (active vitamin D3), he did not respond and continued to complain of bone pain. Several bone resorption markers and bone-specific alkaline phosphatase were also persistently elevated. Therefore, we added denosumab to vitamin D3 supplementation for treatment of excessive bone resorption. Two months after initiation of denosumab, his hip and knee pain was relieved, along with a decrease in serum alkaline phosphatase and some bone resorption markers.ConclusionsAlthough denosumab is not generally an appropriate treatment for acquired Fanconi syndrome, it may be useful for patients who have hypophosphatemic osteomalacia due to adefovir dipivoxil-induced Fanconi syndrome associated with excessive bone resorption. However, clinicians should keep in mind that if denosumab is administered to patients with hypophosphatemic osteomalacia accompanied by excessive bone resorption, adequate vitamin D and/or phosphate supplementation should be done before administration of denosumab.

Evaluating the therapeutic effect of Diallyl disulfide compared to that of Alderonate on Glucocorticoids induced osteoporosis in rats: Biochemical and histomorphometric analysis

Introduction: The prevalence of Glucocorticoids (GC) use made Glucocorticoid induced osteoporosis (GIO) an important form of secondary osteoporosis. Bisphosphonates (Alderonate) are considered as pharmacological agents in prevention and treatment of GIO. Garlic containing Diallyl disulfide (DAS) has received special attention for its beneficial effects as an antioxidant. The present study attempted to evaluate the Alendronate, DAS, and the combination of Alderonate and DAS effect on bone gene expression of RANKL and OPG, serum biochemical parameters [Ca, P, alkaline phosphatase (ALP)] and histological assessment of tibia in animal model of GIO. Material and Method: Fifty pathogen albino male rats were allocated in five groups:Control group(C), Methyl prednisolone group (M), methyl prednisolone Aldenronate group (A) Methyl prednisolone Diallyl disulfide (D), Diallyl disulfide Alendronate Methyl prednisolone (AD) group.Gene expression studies, biochemical parameters, histological and morphometric studies were assessed for all animals. Results: Among the study groups, Methyl prednisolone exposure provoked decrease in OPG (0.26±0.16) increase in RANKL (3.57±.39) gene expression, decrease in serum ALP(85.38±6.3),Ca(6.41±0.89), P(1.9 ±0.35) levels and decrease in trabecular thickness (57.01±23.22).Alderonate and Dially disulfide concomitant administration was shown to increase OPG(2.84 ±0.53)and decrease in RANKL (0.63 ± 0.27) gene expression, increase serum ALP(187.75±24.93),Ca(10.330 ±.69)and P(3.16 ±0.43) levels,increase trabecular thickness(154.7±31.7)(p < 0.001). Conclusion: Diallyl disulfide can add advantage to Alderonate in treatment of glucocorticoid induced osteoprosis.

Rickets severity predicts clinical outcomes in children with X-linked hypophosphatemia: Utility of the radiographic Rickets Severity Score.

The Rickets Severity Score (RSS) was used to evaluate X-linked hypophosphatemic rickets (XLH), a genetic disorder mediated by increased circulating FGF23. The reliability of the RSS was assessed using data from a randomized, phase 2 clinical trial that evaluated the effects of burosumab, a fully human anti-FGF23 monoclonal antibody, in 52 children with XLH ages 5 to 12 years. Bilateral knee and wrist radiographs were obtained at baseline, week 40, and week 64. We evaluated the relationships of the RSS to the Radiographic Global Impression of Change (RGI-C), serum alkaline phosphatase (ALP), height Z-score, 6-minute walk test (6MWT) percent predicted, and the Pediatric Orthopedic Society of North America Pediatric Outcomes Data Collection Instrument (POSNA-PODCI). The RSS showed moderate-to-substantial inter-rater reliability (weighted kappa, 0.45-0.65; Pearson correlation coefficient (r), 0.83-0.89) and substantial intra-rater reliability (weighted Kappa, 0.66; r = 0.91). Baseline RSS correlated with serum ALP (r = 0.47). Baseline RSS identified two subgroups (higher [RSS ≥1.5] and lower RSS [RSS <1.5]) that discriminated between subjects with greater and lesser rachitic disease. Higher RSS was associated with more severe clinical features, including impaired growth (Z-score, -2.12 vs -1.44) and walking ability (6MWT percent predicted, 77% vs 86%), more severe self-reported pain (29.9 [more severe] vs 45.3 [less severe]) and less physical function (29.6 [more severe] vs 40.9 [less severe]). During burosumab treatment, greater reductions in RSS corresponded to higher RGI-C global scores (r = -0.65). Improvements in RSS correlated with decreased serum ALP (r = 0.47). These results show the reliability of the RSS in XLH, and demonstrate that higher RSS values are associated with greater biochemical, clinical, and functional impairments in children with XLH.

Four novel mutations in the ALPL gene in Chinese patients with odonto, childhood, and adult hypophosphatasia.

Hypophosphatasia (HPP) is a rare inherited disorder characterized by defective bone and/or dental mineralization, and decreased serum alkaline phosphatase (ALP) activity. ALPL, the only gene related with HPP, encodes tissue non-specific ALP (TNSALP). Few studies were carried out in ALPL gene mutations in the Chinese population with HPP. The purpose of the present study is to elucidate the clinical and genetic characteristics of HPP in five unrelated Chinese families and two sporadic patients. Ten clinically diagnosed HPP patients from five unrelated Chinese families and two sporadic patients and fifty healthy controls were genetically investigated. All 12 exons and exon–intron boundaries of the ALPL gene were amplified by PCR and directly sequenced. The laboratory and radiological investigations were conducted simultaneously in these HPP ten patients. A 3D model of the TNSALP was used to predict the dominant negative effect of identified missense mutations. Three odonto, three childhood, and four adult types of HPP were clinically diagnosed. Ten mutations were identified in five unrelated Chinese families and two sporadic patients, including eight missense mutations and two frameshift mutations. Of which, four were novel: one frameshift mutation (p.R138Pfsx45); three missense mutations (p.C201R, p.V459A, p.C497S). No identical mutations and any other new ALPL mutations were found in unrelated 50 healthy controls. Our study demonstrated that the ALPL gene mutations are responsible for HPP in these Chinese families. These findings will be useful for clinicians to improve understanding of this heritable bone disorder.

Levetiracetam and lamotrigine effects as mono- and polytherapy on bone mineral density in epileptic patients.

The purpose of this study was to determine the effect of lamotrigine (LTG) and levetiracetam (LEV) as mono- and polytherapy on biochemical markers of bone turnover and bone mineral density in Egyptian adult patients with epilepsy. Forty-eight patients were divided into four groups: two received monotherapy of either LTG or LEV, and the other two groups received polytherapy comprising (valproate [VPA] + LTG or VPA + LEV). Thirty matched healthy participants were included in the study. Participants completed a nutritional and physical activity questionnaire. Biochemical markers of bone and mineral metabolism and bone mineral density of the lumbar spine were measured at baseline and at six months. In the LEV monotherapy group, the bone formation markers showed a significant decrease in serum alkaline phosphatase and serum osteocalcin levels while the bone resorption marker showed a significant increase in urinary deoxypyridinoline levels. After six months of treatment, bone mineral density showed a significant decrease in all treated groups, while among monotherapy groups, this significant decrease was more prevalent in the LEV monotherapy group compared with the LTG monotherapy group. Furthermore, there was significant negative correlation between urinary deoxypyridinoline levels and bone mineral density in the LEV monotherapy group. Using new generation antiepileptics, LEV monotherapies and polytherapy showed harmful effects on bone but LTG did not.

Genetically Engineering a Sheep Model of Hypophosphatasia

Hypophosphatasia (HPP) is a rare inherited disorder that affects the development of bones and teeth. The disease is caused by mutations in the tissue‐nonspecific alkaline phosphatase (TNSALP) gene (ALPL) and accompanied by a highly variable clinical presentation. Although HPP patient studies have advanced our understanding of HPP, as well as documented disease severity, the rarity of the disease combined with a lack of sufficient animal models has significantly delayed mechanistic understanding and therapeutic development. Similar to other human disease models, current HPP models have been engineered virtually exclusively in rodents – specifically mice harboring null or loss‐of‐function mutations. Although useful for modeling some features of HPP, these murine models do not faithfully represent the broad spectrum of human HPP clinical bone, muscle and tooth (odonto‐HPP) phenotypes. However, in sheep both Haversian bone remodeling and tooth development are analogous to humans with sheep TNSALP amino acid sequence sharing 89% identity with human. Thus, our objective was to generate a sheep model of HPP and we posit that ALPL mutation‐specific replacements in the sheep genome using CRISPR/Cas9 will produce a model that accurately phenocopies the bone and tooth pathophysiology of HPP. To genetically engineer a sheep model of HPP, a total of 52 in vitro genetically manipulated embryos were generated targeting the Exon 10 c.1077G&gt;A mutation using CRISPR/Cas9. Three embryos were implanted per recipient ewe and implantations were performed in a total of 17 recipient ewes. Nine ewe pregnancies were confirmed by the measurement of Pregnancy‐Associated Protein on post‐implantation day 35 and further validated by ultrasound. From 9 live newborn lambs, four were heterozygous for our specific point mutation (4/6), one compound heterozygous (1/6) and one homozygous (1/6) for a total mutation efficiency rate of 66.6% (6/9). Similar to HPP patients with the same mutation, mutant sheep have variable clinical expression accompanied with decreased bone formation and mineralization in tails observed by DXA as well as significantly and appropriately decreased serum alkaline phosphatase activity. Additionally, an apparent “shell tooth” phenotype is observed in mutant lambs, further suggesting a mutant exon 10 genotype/phenotype that more completely recapitulates human skeletal and odonto‐HPP.Support or Funding Information1. William Townsend Porter Pre‐doctoral Fellowship2. TAMU/Texas Heart Institute Center for Cell and Organ Biotechnology Innovation Kitchen GrantThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Fertaric Acid Protects from Octylphenol-Related Hepatotoxicity in Rats: Biochemical, Molecular, and Histopathological Studies

ABSTRACTThe endocrine-disrupting chemical 4-tert-octylphenol (OP) can mimic estrogen and testosterone hormones and threaten health; fertaric acid (FA) is a hydroxycinnamic acid found in grapefruit. This study aimed to investigate whether FA has a protective effect on 4-tert-octylphenol–related hepatotoxicity. Thirty male albino rats were divided into 5 equal groups of 6 rats each as follows: control group—administrated orally with 1 ml saline 3 days/week for 4 weeks; corn oil group—administrated orally with 1 ml corn oil 3 days/week for 4 weeks; FA-treated group—administrated orally with FA (45 mg /kg body weight) dissolved in saline 3 days/week for 4 weeks; OP-treated group—administrated orally with OP (40 mg /kg body weight) dissolved in corn oil 3 days/week for 4 weeks; FA + OP–treated group—administrated orally with FA (45 mg /kg body weight) dissolved in saline 3 days/week for 4 weeks then administrated orally with OP (40 mg/kg body weight) dissolved in corn oil 3 days/week for another 4 weeks. The results obtained showed that OP-exposed rats had significant increase in serum aspartate aminotransferase, alanine aminotransferase, γ-glutamyl transferase, lactate dehydrogenase, bilirubin, serum and liver tumor necrosis factor-α, interleukin-1β and malondialdehyde, serum interleukin-6 and interleukin-10 and significant decrease in serum alkaline phosphatase, acid phosphatase, serum and liver superoxide dismutase, glutathione peroxidase, and catalase. OP caused an inhibitory action on the gene expression of liver proteins. Rats treated with FA before OP exposure had near-normal values. In addition, FA prevented the degradation of liver deoxyribonucleic acid (DNA), and DNA reformation occurred. In conclusion, FA protects from dangerous OP-related hepatic effects, and these results were supported by molecular and histological investigations.

Effects of a high dose of microbial phytase and myo-inositol supplementation on growth performance, tibia mineralization, nutrient digestibility, litter moisture content, and foot problems in broiler chickens fed phosphorus-deficient diets

A total of 660 one-day-old Ross 308 broiler chicks were randomly distributed into eleven dietary treatments. Treatments included a maize-soybean meal-based diet with recommended calcium (Ca) and non-phytate phosphorus (nPP) (positive control; PC), an nPP-deficient diet (negative control; NC), NC diets supplemented with different levels of phytase (0, 500, 1,000, 2,000, 3,000, 4,000, 5,000, and 6,000 FTU/kg), a NC diet plus 0.15% myo-inositol, and a NC diet with reduced Ca level (Ca to nPP ratio same as PC). Feeding the NC diet had no effects on birds' body weight (BW), weight gain (WG), feed intake (FI), and feed conversion ratio (FCR), but decreased (P <0.05) tibia P contents, crude protein (CP) digestibility, and serum P, but increased (P <0.05) serum alkaline phosphatase (ALP) activity at 21 d of age. Phytase supplementation at ≥4,000 FTU/kg improved (P <0.05) BW, WG and digestibility of nutrients. Feeding the NC diet resulted in greater (P <0.05) litter moisture content (42 d) and poorer gait score (21 d), but 4,000 and 6,000 FTU/kg phytase returned (P <0.05) these parameters to that of the PC. Supplemental myo-inositol increased (P <0.05) serum total protein, P retention, and decreased (P <0.05) litter moisture at 42 d of age. Feeding the low Ca NC diet increased (P <0.05) serum total protein, ileum Ca, P, and CP digestibility and decreased serum ALP activity, litter moisture and gait score compared to the NC group. In conclusion, phytase in a dose-dependent manner, especially at ≥4,000 FTU/kg levels, was effective in overcoming the negative consequences of NC diets, primarily due to the ability to improve nutrient utilization. In addition, reducing the Ca level or supplementation of inositol of NC diet can correct some the negative effects of feeding a NC diet confirming the negative effect of a wide Ca: P ratio in a P-deficient diet and suggesting that inositol may play a role in the response to phytase addition.

Adult hypophosphatasia caused by ALPL mutation: one pedigree study

An adult patient with hypophosphatasia caused by compound heterozygous mutations in alkaline phosphatase, liver /bone /kidney(ALPL)gene was investigated through comprehensively reviewing the medical history and clinical records of the proband and her family members in order to better understand the disease. The proband and her older sister had mild decreased serum alkaline phosphatase level accompanied with frequently nontraumatic fractures at limbs and all the teeth fell off at the age of 20 and 7, respectively. Both of them carried a missense mutation c. 407G>A(p.Arg136His)in exon 5 and a deletion mutation c. 1318_1320delAAC(p.Asn440del)in exon 12 simultaneously. Other four family members were p. Arg136His mutation carriers and two members were p. Asn440del mutation carriers. We found that p. Asn440del mutation was associated with the oral disorders. In this family, compound heterozygous manifested more serious symptoms, while heterozygous showed relatively mild symptoms. In addition, it is necessary to differentiate it from primary osteoporosis and other diseases of disturbed bone mineralization. (Chin J Endocrinol Metab, 2017, 33: 585-589) Key words: Adult hypophosphatasia; ALPL gene; Compound heterozygous mutations

Enhanced therapeutic benefit of quercetin-loaded phytosome nanoparticles in ovariectomized rats

Quercetin, a dietary flavonol phytoestrogen, has many health benefits but it is poorly absorbed when administered orally. To improve its bioavailability, we prepared quercetin-loaded phytosome nanoparticles (QP) using the thin film hydration method. The prepared nano-formulations were characterized using different techniques. Transmission electron microscopy revealed the homogeneously spherical, well and uniformly dispersed, nano-sized nature of QP. Dynamic light scattering measurements of QP (70 ± 7.44 nm) also confirmed this. Stability of the formed nanoparticles was established via zeta potential determination. The prepared QP exhibited very high encapsulation efficiency (98.4%). The estrogenic activity of QP, concerning inflammation, oxidative stress, bone, lipid profile, blood glucose level and weight gain, was investigated in ovariectomized rat model using 10 and 50 mg/kg/day oral doses for 4 weeks. Treatment with QP showed significant increase in serum calcium, inorganic phosphorus and glutathione content. Whereas, it significantly decreased serum alkaline phosphatase, acid phosphatase, malondialdehyde level, tumor necrosis factor-alpha and glucose level and improved lipid profile. Consequently, the results obtained confirm the superiority of QP over free quercetin at the same doses as a promising hormone replacement therapy.