Alkaline Phosphase Research Articles

Evaluation of hepatoprotective activity of the aqueous extract of Vitis vinifera against acetaminophen-induced liver damage in rats

In this study we evaluated the hepatotoxicity activity of Vitis vinifera in acetaminophen induced liver damage in rats. Five (5) Groups of rats were used for the study, Group 1 received normal saline (1ml /kg, p.o), Group II received acetaminophen (300mg/kg, i.p), Group III received (300mg/kg, i.p) acetaminophen and (150mg/kg, o.p) Vitis vinifera juice, Group IV received ( 600mg/kg, i.p) acetaminophen and (300mg/kg, o.p) Vitis vinifera juice while Group V received ( 600mg/kg, i.p) acetaminophen and Silymarin (25mg/kg, o.p) simultaneously once daily for four (4) days. Acute toxicity study and histopathological analysis were carried out. The LD 50 was found to greater than 4000mg/kg body weight. The aqueous extract of Vitis vinifera significantly (P< 0.05*) reduced the hepatic enzymes from aspartase transaminase ( AST ) (415 ± 0.2* to 94.5 ± 0.8*) alanine transaminase (ALT) (132 ± 2.2 * to 60.5 ± 0.2*) and alkaline phosphase ALP (118± 0.6 * to 87.5 ± 2.1*) while there were no significant changes in the total bilirubin (TB) and direct bilirubin levels comparable to the reference drug. These findings confirm the histopathology study which indicated no adverse effects on the liver. The aqueous extract of the Vitis vinifera possesses hepatoprotective activity against acetaminophen induced liver damage, therefore further studies should be carried out to determine the exact mechanism of action as well as phytochemicals responsible for these activities.

QUALITY OF LIFE AND RESILIENCE IN EARLY SURGICAL TREATMENT OF CHERUBISM IN A PEDIATRIC PATIENT: CASE REPORT

This study reports the case of an eight-year-old child who presented facial asymmetry due to increased bilateral volume in the face. Computed tomography presented bilateral multilocular osteolytic solutions in the mandible and maxilla, with the expansion and thinning of the cortical bones. Laboratory tests released figures of calcium, alkaline phosphase, and emonium. An incisional biopsy was performed, and histopathological examination revealed features suggestive of a giant cell lesion. After correlating the history with the clinical, radiographic and histological findings, the diagnosis of cherubism was reached. The impact of facial treatment deformity on quality of life, evaluated through a well-recognized treatment (PedsQL), led to early treatment with curettage and bone surgery. After 6 months, the patient shows an improved facial symmetry and quality of life. This study reports the case of an eight-year-old child who presented facial asymmetry due to increased bilateral volume in the face. Computed tomography presented bilateral multilocular osteolytic solutions in the mandible and maxilla, with the expansion and thinning of the cortical bones. Laboratory tests released figures of calcium, alkaline phosphase, and emonium. An incisional biopsy was performed, and histopathological examination revealed features suggestive of a giant cell lesion. After correlating the history with the clinical, radiographic and histological findings, the diagnosis of cherubism was reached. The impact of facial treatment deformity on quality of life, evaluated through a well-recognized treatment (PedsQL), led to early treatment with curettage and bone surgery. After 6 months, the patient shows an improved facial symmetry and quality of life.

Serum Osteocalcin, P1NP, Alkaline Phosphase, and CrossLaps in Humans.The relationship with body mass index

This is a clinical study on 56 subjects included in normal weight (NW) group (N=17), overweight (OW) group (N=19) and grade I obese (O) group (N=20), based on BMI (Body Mass Index) values: NW group had a mean BMI of 22.2 � 2.14 kg/sqm, OW group had a BMI of 25.89 � 1.04 kg/sqm, and O group had an average BMI of 32.2 � 2.09 kg/sqm (p-value NW-OW, NW-O, respective OW-O groups was p[0.0005). The 3 groups were similar as age (p-value NW-OW groups = 0.7, between NW- O groups = 0.8, respective between OW - O group = 0.7). The circulating bone formation (osteocalcin, P1NP alkaline phosphatase) and resorption profile (CrossLaps) indicated no statistical significant difference between groups while the coefficient of regression r between each biochemical bone marker and BMI in every BMI group exceeded the value of p]0.05. All the 3 groups had a mean value of 25-hydroxycholecalciferol in deficiency ranges ([ 30 ng/mL, normal recommended values are above 30 ng/mL) without significant differences regarding BMI groups, except for obese group when compare to the other two groups. No secondary hyperparathyroidism was associated in any group despite low vitamin D levels. Based on our observation, bone turnover biochemical markers are not influenced by BMI.

Protocol for a randomized controlled trial to compare bone-loading exercises with risedronate for preventing bone loss in osteopenic postmenopausal women.

BackgroundIn the United States, over 34 million American post-menopausal women have low bone mass (osteopenia) which increases their risk of osteoporosis and fractures. Calcium, vitamin D and exercise are recommended for prevention of osteoporosis, and bisphosphonates (BPs) are prescribed in women with osteoporosis. BPs may also be prescribed for women with low bone mass, but are more controversial due to the potential for adverse effects with long-term use. A bone loading exercise program (high-impact weight bearing and resistance training) promotes bone strength by preserving bone mineral density (BMD), improving bone structure, and by promoting bone formation at sites of mechanical stress.Methods/DesignThe sample for this study will be 309 women with low bone mass who are within 5 years post-menopause. Subjects are stratified by exercise history (≥2 high intensity exercise sessions per week; < 2 sessions per week) and randomized to a control or one of two treatment groups: 1) calcium + vitamin D (CaD) alone (Control); 2) a BP plus CaD (Risedronate); or 3) a bone loading exercise program plus CaD (Exercise). After 12 months of treatment, changes in bone structure, BMD, and bone turnover will be compared in the 3 groups. Primary outcomes for the study are bone structure measures (Bone Strength Index [BSI] at the tibia and Hip Structural Analysis [HSA] scores). Secondary outcomes are BMD at the hip and spine and serum biomarkers of bone formation (alkaline phosphase, AlkphaseB) and resorption (Serum N-terminal telopeptide, NTx). Our central hypothesis is that improvements in bone strength will be greater in subjects randomized to the Exercise group compared to subjects in either Control or Risedronate groups.DiscussionOur research aims to decrease the risk of osteoporotic fractures by improving bone strength in women with low bone mass (pre-osteoporotic) during their first 5 years’ post-menopause, a time of rapid and significant bone loss. Results of this study could be used in developing a clinical management pathway for women with low bone mass at their peak period of bone loss that would involve lifestyle modifications such as exercises prior to medications such as BPs.Trial registrationClinicaltrials.gov NCT02186600. Initial registration: 7/7/2014.Electronic supplementary materialThe online version of this article (doi:10.1186/s12905-016-0339-x) contains supplementary material, which is available to authorized users.

The effects of liraglutide on the proliferation and differentiation of mouse pre-osteoblasts MC3T3-E1

Objective To investigate the effects of liraglutide on the proliferation and differentiation of mouse pre-osteoblasts MC3T3-E1 exposed to higher glucose concentration. Methods MC3T3-E1 cells were cultured and divided into control group and liraglutide group. In liraglutide group, cells were treated with different liraglutide concentrations (10-9 mol/L, 10-8 mol/L, and 10-7 mol/L, respectively) for 48 hours. Cell proliferation was tested with CCK-8. The mRNA expressions of type Ⅰ collagen (COL-Ⅰ), osteopontin (OPN), and alkaline phosphase (ALP) were detected using semi-quantitative RT-PCR. Results (1)Compared to control group, the proliferation rate of different liraglutide concentration groups (10-9 mol/L, 10-8 mol/L, and 10-7 mol/L) increased significantly (all P<0.05), the proliferation rate was the highest in 10-8mol/L liraglutide group. (2)The expression of COL-Ⅰ, OPN, and ALP mRNA in liraglutide groups were higher than those in control group (all P<0.05). The optimal concentration of liraglutide was 10-8 mol/L. Conclusion Liraglutide within a certain concentration range may improve the proliferation and differentiation of mouse pre-osteoblasts MC3T3-E1. (Chin J Endocrinol Metab, 2016, 32: 232-235) Key words: Glucagon- like peptide-1 analogue; Liraglutide; Osteoblast; Proliferation; Differentiation

Gene Silencing of Chordin Improves BMP-2 Effects on Osteogenic Differentiation of Human Adipose Tissue-Derived Stromal Cells

Although bone morphogenic protein (BMP)-2 is known to potently induce osteogenic differentiation of human mesenchymal stem cells, strong individual differences have been reported. In part, this is due to internal antagonists of BMP-2 for example, noggin and chordin, secreted by differentiating cells. This enabling study was performed to prove the hypothesis that osteogenic effects of BMP-2 can be improved by transient nonviral gene silencing of chordin. We investigated the effect of siRNA against chordin on osteogenic differentiation in human adipose tissue-derived stromal cells (hASC). Cells of two different donors were isolated after liposuction and proliferated for passage 4 or 5. On seeding, hASCs were transfected with siRNA using a commercial liposomal transfection reagent. Subsequently, cells were differentiated in the presence or absence of BMP-2 (100 ng/mL). Noncoding siRNA as well as siRNA against noggin served as a control. Osteogenic differentiation of hASC was determined by alkaline phosphase (ALP) activity and matrix mineralization. ALP activity of hASC treated with siRNA against chordin was increased for cells of both donors. In contrast, silencing of noggin had no effect in any of the donors. In combination with BMP-2, silencing of either chordin or noggin showed strongly improved ALP activity compared with the control group that was also supplemented with BMP-2. Mineralization was observed to start earlier in groups that received siRNA against chordin or noggin and showed increased amounts of incorporated calcium on day 15 compared with the control groups. Silencing chordin in hASCs was successful to increase BMP-2 effects on osteogenic differentiation in both donors, while effects of noggin silencing were reliably observed only in one of the two investigated donors. In contrast to noggin silencing, chordin silencing also increased osteogenic differentiation without supplemented BMP-2.

Protective Effect of Nigella sativa Oil against Methotrexate Induced Hepatotoxicity in Children with Acute Lymphoblastic Leukemia

Background: Acute Lymphoblastic Leukemia (ALL) is the most common childhood malignancies representing about one third of all pediatric cancers. Adding methotrexate to leukemia treatment protocols has been associated with an increased survival rate in children with ALL. The efficacy of this agent is often limited by its toxicity which can be reduced if supplemented with anti-oxidants. Nigella sativa has antioxidant property through different mechanisms. Objective: The aim of this work was to study the role of Nigella sativa oil in the protection against hepatotoxicity induced by methotrexate therapy in children with ALL and the impact on the treatment outcome. Patients and methods: The present study was conducted in the period between July 2010 and July 2013 on 40 children with newly diagnosed ALL including 28 males and 12 females, with mean age value of 9.17 ± 3.81 years and they were divided into 20 patients of ALL under methotrexate therapy included in ALL treatment protocol, delayed leukovorin rescue (10 mg/m2 orally or IV every 6 hours for five doses beginning 48 hours after start of methotrexate infusion and Nigella sativa oil in form of soft gelatin capsule 450 mg in dose of 80 mg/kg/day on three divided doses for one week after each methotrexate dose (Group II) and 20 patients of ALL under methotrexate therapy included in ALL treatment protocol, delayed leukovorin rescue (10 mg/m2 orally or IV every 6 hours for five doses beginning 48 hours after start of methotrexate infusion and placebo for one week after each methotrexate dose (Group III). This study also included 20 healthy children as a control group (11 males and 9 females) with their mean age value of 9.1+ 2.9 (Group I). All patients included in the study were subjected to the following investigations: Complete blood picture, bone marrow aspiration, cytochemistry, immunophenotyping and liver function testes. Results: There were no significant difference in serum bilirubin, alanine aminotransferase, aspartate aminotransferase, alkaline phosphase levels and prothrombin time between group II and group III but there was significant difference between group II and group III compared to controls. There was no significant difference in total protein, albumin, globulin levels, and albumin globulin ratio between studied groups. There were non-significant increase in total, direct and indirect serum bilirubin, serum ALT, AST, and alkaline phosphatase levels and prothrombin time in group II after methotrexate and Nigella sativa oil therapy but there was significant increase in group III after treatment with methotrexate and placebo with significant difference between group II and III after therapy. There were significant differences in overall and disease free survival between group II and group III. Conclusion: Oral administration of Nigella sativa oil in leukemic children can prevent MTX hepatotoxicity and improved survival in patients with ALL. Recommendations: Nigella sativa oil is recommended adjuvant drug as hepatoprotective agent in patients with ALL who received methotrexate therapy.

Influence of surface topography on osteoblast response to fibronectin coated calcium phosphate thin films

The ability to engineer biomaterial surfaces that are capable of a dynamic interaction with cells and tissues is central to the development of medical implants with improved functionality. An important consideration in this regard is the role played by the extracellular proteins that bind to an implant surface in vivo. Deliberate use of an ad-layer of such proteins on an implant surface has been observed to guide and direct cell response. However, the role that changes in surface topography might play in determining the nature of this cell–protein–surface interaction has not been investigated in detail. In this study, calcium phosphate (CaP) thin films have been deposited onto substrates with varying topography such that this is reflected in the (conformal) CaP surface features. A fibronectin (FN) ad-layer was then deposited from solution onto each surface and the response of MG63 osteoblast-like cells investigated. The results revealed that in all cases, the presence of the adsorbed FN layer on the CaP thin films improved MG63 cell adhesion, proliferation and promoted early onset differentiation. Moreover, the nature and scale of the response were shown to be influenced by the underlying CaP surface topography. Specifically, MG63 cell on FN-coated CaP thin films with regular topographical features in the nanometer range showed statistically significant differences in focal adhesion assembly, osteocalcin expression and alkaline phosphase activity compared to CaP thin films that lacked these topographical features. As such, these data indicate that surface topography can be used to further influence cell adhesion and downstream differentiation by enhancing the effects of a surface adsorbed FN layer.

Estrogenic properties of traditional Cameroonian medicinal plants

Medicinal plants of tropical and subtropical areas of Cameroon, which are traditionally used to treat female ailments, were investigated towards potential estrogenic activities. In a first series of screening experiments in the yeast estrogen receptor assay on 33 extracts from 18 plants estrogenic activities in the ethyl acetate extracts of the stem bark of Millettia conraui and the stem bark of Millettia drastica, as well as the methanol extracts of the leaves of Bridelia ferruginea, the roots of Pseudarthria hookeri and the roots of Nauclea latifolia could be shown. These results could be verified by alkaline phosphase induction in Ishikawa cells at 10 and 100mg/ml doses, except for the extract of Bridelia ferruginea which was toxic for Ishikawa cells. All stimulatory effects were inhibited by a combined treatment of cells with the pure antiestrogen fulvestrant. In conclusion, the extracts of Millettia conraui, Millettia drastica, Pseudarthria hookeri and Nauclea latifolia may contain secondary metabolites endowed with estrogenic activity. In another experiment, the ethyl acetate extract of the stem bark of Erythrina lysistemon also proved to be estrogenic in the two in vitro assays described above. These results prompted us to investigate the estrogenic activity of this extract in vivo in young ovariectomized female Wistar rats after a 7-day treatment. Preliminary results obtained to date concern the extract of Erythrina lysistemon, which was evaluated through the proliferative status of target sex organs such as uterus and vagina. The oral administration of 200mg/kg BW/d of Erythrina lysistemon extract in comparison to untreated ovariectomized rats significantly increased the vaginal epithelial height by 47% and induced a weak increase of uterine epithelial height around 7%. Both effects were not as pronounced as those elicited in positive control of 100µg/kg BW/d of ethinylestradiol given orally. Overall our results suggest that the extract of Erythrina lysistemon contains secondary metabolites endowed with estrogenic activity.

Phosphorylation is a regulatory mechanism in apolipoprotein B mRNA editing.

The editing of apolipoprotein B (apoB) mRNA is under tissue-specific, developmental and metabolic regulation. We found that multiple protein kinase inhibitors or activators increased apoB mRNA editing up to 2.5-fold in Caco-2 cells and 3-8-fold in McA7777 and FAO rat cells respectively. The phosphorylation-agent-induced modulation is independent of the apolipoprotein B editing catalytic subunit 1 (APOBEC-1) and of apoB mRNA expression levels, indicating the involvement of a protein modification, such as phosphorylation, regulating the cellular editing of apoB mRNA. Transient expression of protein kinase C-&theta; more than doubled apoB mRNA editing in FAO cells. Chronic exposure to ethanol, a treatment known to increase the expression of protein kinases and to change protein phosphorylation status, increased apoB mRNA editing in FAO cells up to 2.5-fold without increasing the mRNA abundance of APOBEC-1. The elimination of potential phosphorylation sites 47 and 72 of human APOBEC-1 decreased its activity to approx. one-eighth of control levels by a Ser(47)-->Ala mutation, but more than doubled the activity by a Ser(72)-->Ala mutation. The activity modulation was reversed by a Ser-->Asp mutation at sites 47 and 72, which introduced a phosphorylation-like carbonic acid group. Both human APOBEC-1 dephosphorylated by alkaline phosphase and the Ser(47,72)-to-alanine double mutant protein demonstrated a shifted isoelectric focusing pattern compared with the wild type, indicating phosphorylation at these sites. Taken together, these results suggest that phosphorylation might be an important mechanism in the regulation of apoB mRNA editing.

Phosphorylation is a regulatory mechanism in apolipoprotein B mRNA editing

The editing of apolipoprotein B (apoB) mRNA is under tissue-specific, developmental and metabolic regulation. We found that multiple protein kinase inhibitors or activators increased apoB mRNA editing up to 2.5-fold in Caco-2 cells and 3–8-fold in McA7777 and FAO rat cells respectively. The phosphorylation-agent-induced modulation is independent of the apolipoprotein B editing catalytic subunit 1 (APOBEC-1) and of apoB mRNA expression levels, indicating the involvement of a protein modification, such as phosphorylation, regulating the cellular editing of apoB mRNA. Transient expression of protein kinase C-θ more than doubled apoB mRNA editing in FAO cells. Chronic exposure to ethanol, a treatment known to increase the expression of protein kinases and to change protein phosphorylation status, increased apoB mRNA editing in FAO cells up to 2.5-fold without increasing the mRNA abundance of APOBEC-1. The elimination of potential phosphorylation sites 47 and 72 of human APOBEC-1 decreased its activity to approx. one-eighth of control levels by a Ser47 → Ala mutation, but more than doubled the activity by a Ser72 → Ala mutation. The activity modulation was reversed by a Ser → Asp mutation at sites 47 and 72, which introduced a phosphorylation-like carbonic acid group. Both human APOBEC-1 dephosphorylated by alkaline phosphase and the Ser47,72-to-alanine double mutant protein demonstrated a shifted isoelectric focusing pattern compared with the wild type, indicating phosphorylation at these sites. Taken together, these results suggest that phosphorylation might be an important mechanism in the regulation of apoB mRNA editing.

Hypophosphatemic rickets/osteomalacia in linear sebaceous nevus syndrome: A variant of tumor-induced osteomalacia

A severe form of vitamin D-resistant rickets is associated with the linear sebaceous nevus syndrome. We investigated the pathophysiology underlying defective bone mineralization in two individuals and examined the effects of 1,25-dihydroxyvitamin D (1,25(OH)2D, calcitriol) therapy on the clinical and biochemical abnormalities. Both patients had fasting hypophosphatemia, markedly diminished TmP/GFR, and elevated alkaline phosphase activity in the presence of normocalcemia. Before treatment with calcitriol, serum 1,25(OH)2D concentrations were reduced but serum 25-hydroxyvitamin D (25(OH)D) concentrations were normal. Administration of calcitriol increased serum 1,25(OH)2D concentrations and led to an increase in TmP/GFR and serum phosphorus levels and to a decrease in alkaline phosphatase activity. However, the renal tubular maximum for reabsorption of inorganic phosphate, normalized according to glomerular filtration rate, and serum phosphorus levels remained abnormally low even in the patient who also received phosphate supplementation. Bone histomorphologic studies in the adult patient showed extreme osteomalacia, which partially improved with calcitriol. These data demonstrate that the putative skin lesion-derived factor results in both a renal tubular defect in phosphate reabsorption and in 1,25-(OH)2 D deficiency. The vitamin D-resistant rickets of linear sebaceous nevus syndrome is a variant of tumor-induced osteomalacia.

Effect of 1,25-dihydroxycholecalciferol on the duodenal villi and alkaline phosphatase in the developing chick embryo.

Administration of 1,25-(OH)2D3 to developing 14-day chick embryo gave precocious induction of alkaline phosphatase in 20-day chick embryonic duodenum. 1,25-(OH)-2D3-induced alkaline phosphase involved in changes in Km and Vmax values. Furthermore, polyacrylamide gel disc electrophoresis of n-butanol-solubilized alkaline phosphatase from control and 1,25-(OH)2D3-treated chick embryonic duodenum revealed that 1,25-(OH)2D3 involved the transformation of neuraminidase-resistant fast migrating form to the neuraminidase-sensitive faster migrating one. Scanning electron microscopic data showed that the injection of 1,25-(OH)2D3 stimulated the elongation of duodenal microvilli, although there was no effect on the duodenal absorptive epithelial cell height.

Separation, properties, localization and nycthemeral variations of a DNAse, a phosphodiesterase, a RNAse and an alkaline phosphase from the hepatopancreas of Palaemon serratus (Crustacea Natantia)

1. 1. We have identified in crude extracts of shrimp hepatopancreas ( Palaemon serratus) deoxyribonuclease, ribonuclease, phosphodiesterase and alkaline phosphatase. 2. 2. The four enzymatic activities were characterized by their optima pH and the molecular weight of the enzymes. 3. 3. DNAse and phosphodiesterase were separated and further characterized for their cleavage sites and their ionic requirements. 4. 4. Amylases, proteases, phosphatases and phosphodiesterases vary synchronously on a 24 hr period with a two peak pattern of activity. 5. 5. The maximum activities of RNAse and DNAse occur assynchronously. The possible metabolic role of these latter enzymes is discussed.

Significance of serum enzymatic activities in genitourinary tract malignancy (author's transl)

Activities of serum lactate dehydrogenase, leucine aminopeptidase, aldolase and alkaline phosphatase were estimated at the follow-up of 54 cases of genitourinary tract malignancy (renal cell carcinoma 15, renal pelvic and ureteral carcinoma 10, bladder cancer 23. testicular malignancy 6).The following conclusions have been obtained.1. In renal cell carcinoma, the measurement of leucine aminopeptidase was useful at the preoperative diagnosis and the follow-up. Elevated activity of leucine aminopeptidase was noted in the cases of advanced stage.2. The combined evaluation of both leucine aminopeptidase and lactate dehydrogenase might be of some value in terms of the diagnosis and prognosis of renal pelvic and ureteral carcinoma.3. The estimation of lactate dehydrogenase along with alkaline phosphatase and leucine aminopeptidase activities was of some help in the preoperative diagnosis and follow up of bladder cancer.4. At the time of liver metastasis, the value of leucine aminopeptidase was elevated along with alkaline phosphase activity in renal cell carcinoma, while the elevated activity of lactate dehydrogenase was observed in bladder cancer.

A micromethad for the determination of inorganic phosphorus and Phosphatase

A micro-colorimetric method for the determination of inorganic phosphorus in the biological fluids and serum phosphatase is described. Its use as a rapid and convenient method for estimating the serum inorganic phosphorus and alkaline phosphase of the rachitic children is of particular value.