Alkaline Phosphatase Fraction Research Articles

The Evaluation of Bone Regeneration Following Socket Preservation with Concentrated Growth Factor (CGF) and Poly Lactic-Co-Glycolic Acid (PLGA) Scaffold in Rabbits

Various grafting materials are utilised to facilitate regeneration. There is currently a paradigm shift towards applying poly lactic-co-glycolic acid (PLGA), which is regarded as an excellent scaffold for tissue engineering. Concentrated growth factor (CGF) has also been reported to promote wound healing. Nevertheless, the role of PLGA microspheres as a substitute for bone graft material with CGF in bone regeneration remains unclear. This study was designed to evaluate the effect of CGF with PLGA on bone formation and the expression of alkaline phosphatase (ALP) following socket preservation. PLGA microspheres were prepared using double solvent evaporation method and observed under scanning electron microscopy (SEM). A 6 mL of rabbit’s blood was collected from the marginal ear vein and centrifuged to obtain CGF. Blood was also collected for ALP assessment from 24 New Zealand White (NZW) male rabbits subjected to the first upper left premolar extraction. Sockets were filled with CGF, PLGA, CGF+PLGA or left empty and observed with microscopic computed tomography (micro-CT) at four weeks and eight weeks. The SEM image revealed a spherical shape with interconnected pores on the surface of the PLGA particles. Repeated measures ANOVA were used to evaluate the effect of time and treatment (p < 0.05) with significant differences in bone width, height, volume, volume fraction and expression of ALP was observed with CGF+PLGA. Both CGF and PLGA have the potential as the alternative grafting materials and this study could serve as an ideal benchmark for future investigations on the role of CGF+PLGA in bone regeneration enhancement.

USEFULNESS OF THERMO LABILE ALKALINE PHOSPHATASE AS A BIOMARKER IN DIAGNOSIS OF POST-MENOPAUSAL OSTEOPOROSIS

Background: Osteoporosis is characterized by demineralization of bones and it is a complication of women in post-menopausal life. Bone alkaline phosphatase is an isoenzyme of alkaline phosphatase which helps in the mineralization of bones and it is heat labile in nature. Its measurement facilitates the clinical diagnosis of Osteoporosis. Objectives: To assess the status of total alkaline phosphatase and heat-labile isoenzyme fraction (bone fraction) in serum of pre and post-menopausal women and to relate the findings to the clinical condition /final diagnosis to make out the correlation between the pre and postmenopausal women. Methodology:A total of 70 subjects, consisting of 35 pre-menopausal women (38.11 ± 4.3 years of age) and 35 post-menopausal women (54.40 ± 4.6 years) were constituted for the study. Their total alkaline phosphatase was measured directly and the heat stable alkaline phosphatase was measured after heating the serum at 56℃ for 10 minutes. Spectrophotometric P- Nitro-Phenyl Phosphate method was followed for the estimation. Heat labile alkaline phosphatase levels were obtained by subtracting the heat stable fraction from total Alkaline phosphatase. Result: The measured serum total alkaline phosphatase level was slightly increased in post menopause compared to premenopausal women but it was not statistically significant. The levels of heat-labile Alkaline phosphatase showed significant difference between pre and post-menopausal women by ***: p < 0.01: ***: p < 0.001 respectively. Conclusion: Heat labile alkaline phosphatase can be used as a biomarker to diagnose Osteoporosis in post-menopausal women.

Bioinformatical analysis of the level of osteomarkers in the gingival fluid in systemic osteoporosis and local osteo-destructive processes in the dentoalveolar region

The informational value of assessing biochemical markers in the gingival fluid of patients with various systemic and local changes in bone tissue was analyzed in a sample of 57 patients and 15 healthy people. The patients were differentiated into 3 groups: 1) 20 patients with ulcerative colitis and steroid osteoporosis due to glucocorticoid therapy; 2) 22 patients diagnosed with cancer of the mucous membrane of the floor of the mouth with erosions of the cortical bone or dental socket by a primary tumor; 3) 15 patients with moderate dental peri-implantitis. Gingival fluid was used as a test material. The content of cathepsin K, tartrate-resistant acid phosphatase, and bone fraction of alkaline phosphatase was determined. In steroidal osteoporosis, the cathepsin K level in the gingival fluid was found to be statistically significantly increased compared to other markers under study. In group 2, with tumor osteodestruction in the gingival fluid, a 10-fold increase in the concentration of cathepsin K and tartrate-resistant acid phosphatase was observed against the background of a moderate increase in the bone fraction of alkaline phosphatase content. In osteodestruction associated with inflammatory dentoalveolar processes, a moderate increase in the concentration of cathepsin K and tartrate-resistant acid phosphatase was observed. Thus, the determination of osteomarkers in the gingival fluid can be considered informative in terms of distinguishing between systemic and local changes in the structure of bone tissue.

Cause-and-effect relationship between thyroid and liver diseases

Thyroid gland (TG) and the liver are in a complex relationship in both physiological and pathological conditions. Thyroid hormones accelerate metabolic processes, intensify the synthesis of proteins and vitamins, play an important role in the development and differentiation of all cells, including hepatocytes. In addition to the central role in the deiodination of thyroid hormones with the formation of their more active and inactivated forms, the liver also carries out their transport. Dysfunction of TG can lead to changes in liver function, and in liver diseases, abnormalities in the metabolism of thyroid hormones can occur. Most often, liver pathology in diseases of TG is manifested by an increase in the serum activity of enzymes of cytolysis and/or cholestasis. Changes in liver function tests are often observed in patients with thyrotoxicosis. They are based on oxidative stress or cholestasis. The increased activity of osteoblasts in hyperthyroidism leads to an increase in the bone fraction of alkaline phosphatase, which must be taken into account in the differential diagnosis. Hepatotoxicity of thyreostatic drugs is relatively common, ranging from minimal hepatocellular damage to fulminant liver failure. In the case of hypothyroidism, the pathophysiological mechanisms are mainly represented by lipid metabolism disorders leading to fatty degeneration. It should be remembered that severe hypothyroidism can be manifested by hyperammonemia and edematous-ascitic syndrome, requiring differential diagnosis with liver failure. Treatment of liver pathology in TG diseases includes normalization of thyroid status, and in cases of drug hepatitis – temporary withdrawal of a potentially hepatotoxic drug. The data on the association of hypothyroidism and non-alcoholic fatty liver disease in the aspect of developing new therapies are very interesting.

Корекція порушень ремоделювання кісткової тканини при довготривалій кортикостероїдній терапії

Актуальність. При тяжких дерматозах зазвичай призначаються препарати кортикостероїдних гормонів досить тривалими курсами і навіть довічно, у зв’язку з чим діагностика виникнення побічних ефектів від тривалого використання системних глюкокортикостероїдів (ГКС), зокрема порушень кісткового обміну, а також їх профілактика є важливою складовою комплексної терапії тяжких дерматозів. Тому розробка методів ранньої діагностики, лікування і профілактики появи порушень ремоделювання кісткової тканини під впливом тривалої глюкокортикостероїдної терапії у хворих на тяжкі дерматози вкрай актуальна для сучасної дерматології. Мета дослідження — підвищення ефективності та безпечності лікування хворих на тяжкі дерматози шляхом запобігання або мінімізації розвитку побічних ефектів лікування глюкокортикостероїдними гормонами. Матеріали та методи. У дослідження було залучено 134 хворих на тяжкі дерматози (акантолітична пухирчатка, бульозний пемфігоїд, дерматоз Дюрінга, еритродермія тощо). Використовувалися клініко-анамнестичні, біохімічні, цитологічні, імуноферментні, інструментальні та статистичні методи дослідження. Результати. У хворих у результаті тривалої системної глюкокортикостероїдної терапії виявлялося вірогідне зниження кісткової фракції лужної фосфатази у 2,1 раза, рівня остеокальцину — у 6,6 раза і підвищення рівня С-кінцевого телопептиду у 2,2 раза. За даними ультразвукової денситометрії, остеопенію було виявлено у 35,5 %, остеопороз — у 64,5 % пацієнтів. Для корекції ремоделювання кісткової тканини призначалися препарати кальцію з вітаміном D3 та антирезорбтивний засіб. У результаті відзначалося підвищення зазначених показників, що свідчило про відновлення процесів остеосинтезу та нормалізацію процесів резорбції кісткової тканини, що підтверджувалося підвищенням мінеральної щільності кісткової тканини в 1,8 раза. Для запобігання розвитку порушень процесів ремоделювання кісткової тканини в комплексному лікуванні призначалися препарати антирезорбтивної дії, кальцію і вітаміну D3 в поєднанні із системними ГКС протягом усього курсу терапії. Висновки. Проведене дослідження дозволило підвищити ефективність та безпеку терапії хворих на тяжкі дерматози шляхом запобігання або мінімізації розвитку побічних ефектів лікування глюкокортикостероїдами.

The Dual Effects of Reactive Oxygen Species on the Mandibular Alveolar Bone Formation in SOD1 Knockout Mice: Promotion or Inhibition.

The status of reactive oxygen species (ROS) correlates closely with the normal development of the oral and maxillofacial tissues. Oxidative stress caused by ROS accumulation not only affects the development of enamel and dentin but also causes pathological changes in periodontal tissues (periodontal ligament and alveolar bone) that surround the root of the tooth. Although previous studies have shown that ROS accumulation plays a pathologic role in some oral and maxillofacial tissues, the effects of ROS on alveolar bone development remain unclear. In this study, we focused on mandibular alveolar bone development of mice deficient in superoxide dismutase1 (SOD1). Analyses were performed using microcomputerized tomography (micro-CT), TRAP staining, immunohistochemical (IHC) staining, and enzyme-linked immunosorbent assay (ELISA). We found for the first time that slightly higher ROS in mandibular alveolar bone of SOD1(-/-) mice at early ages (2-4 months) caused a distinct enlargement in bone size and increased bone volume fraction (BV/TV), trabecular thickness (Tb.Th), and expression of alkaline phosphatase (ALP), Runt-related transcription factor 2 (Runx2), and osteopontin (OPN). With ROS accumulation to oxidative stress level, increased trabecular bone separation (Tb.Sp) and decreased expression of ALP, Runx2, and OPN were found in SOD1(-/-) mice at 6 months. Additionally, dosing with N-acetylcysteine (NAC) effectively mitigated bone loss and normalized expression of ALP, Runx2, and OPN. These results indicate that redox imbalance caused by SOD1 deficiency has dual effects (promotion or inhibition) on mandibular alveolar bone development, which is closely related to the concentration of ROS and the stage of growth. We present a valuable model here for investigating the effects of ROS on mandibular alveolar bone formation and highlight important roles of ROS in regulating tissue development and pathological states, illustrating the complexity of the redox signal.

Diagnostic value of biomarkers of bone metabolism in the gingival fluid in inflammatory-destructive and tumor oral pathology

The aim of the work was to determine the informativeness of the assessment of biochemical markers in the gingival fluid with destructive bone changes in the peri-implantation area, primary tumor lesion of the alveolar processes of the jaws and metastatic destruction of the jaw bone tissue in cancer of the large salivary glands. The study included 51 patients aged 30 to 75 years of both sexes. Depending on the genesis of destruction of the bone tissue of the lower jaw, three groups of patients were identified. Group 1 - 20 patients with peri-implantitis. Group 2 - 22 patients diagnosed with cancer of the oral floor mucosa (ICD C04) with erosion of the cortical bone or dental socket by a primary tumor. Group 3 - 9 patients with adenocarcinoma of the large salivary glands with metastatic lesions of the lower jaw. 15 people were practically healthy persons without periodontal pathology. The test material was gingival fluid or exudate from peri-implantation pockets. The content of cathepsin K, tartrate-resistant acid phosphatase (TAP) was determined by the enzyme immunoassay, and the bone fraction of alkaline phosphatase (BAP) was determined by the kinetic method. As a result, it was found that in group 1 in the exudate of the peri-implantation sulcus, the content of cathepsin K increased in comparison with the control by 2.5 times (p<0.05), TAP by 1.6 times (p<0.05). in groups 1 and 2, there was a moderate increase in the production of activated osteoblasts of KAP. In cancer of the mucous membrane of the floor of the mouth, a tenfold increase in the concentration of cathepsin K and TAP in the gingival fluid was observed. With metastatic lesions of the jaw in patients with adenocarcinoma of the large salivary glands, along with a multiple increase in the concentration of cathepsin K and TAP in the gingival fluid, there was a twenty-fold increase in the bone formation marker BAP, which indicated a mixed type of metastases and was a differential diagnostic criterion for metastatic lesion of bone tissue in comparison with the primary metastatic lesion of bone tissue. Thus, biochemical markers are sensitive indicators of remodeling of bone tissue both in local inflammatory and destructive processes in peri-implantitis and in primary and metastatic tumor lesions of the jaws.

2619 Persistently Elevated Intestinal Alkaline Phosphatase in an Individual With AB Blood Group: A Rare Presentation

INTRODUCTION: Elevated alkaline phosphatase (ALP) is seen in hepatobiliary and bone diseases. However, elevated intestinal ALP has been reported in the literature in association with blood group B or O, whose clinical significance is not known. We present a rare case of persistently elevated intestinal ALP in an individual with blood group AB. CASE DESCRIPTION/METHODS: A 56-year-old female presented to our ambulatory clinic to establish health care. Her medical history included peptic ulcer disease and dyslipidemia. She underwent appendectomy in childhood and cholecystectomy around 20 years ago. she denied smoking, alcohol consumption or use of recreational drugs. Family history was significant for type 1 DM in mother and brother. Her only medication included ranitidine. General and systemic examination was normal. Her initial labs were normal except elevated alkaline phosphatase of 359 units/liter. The patient was seen by the gastroenterology team. Further laboratory workup including viral hepatitis, metabolic and autoimmune markers of liver disease were normal. Gamma-glutamyltransferase (GGT), Vitamin D and parathyroid profile were normal. Ultrasound abdomen did not show any significant lesion in the liver or biliary tree. As no evidence of liver disease was noted isoenzyme electrophoresis of ALP was ordered which showed elevated intestinal ALP fraction of 64% (normal 1–24%). Patient blood group was tested which showed group AB. On further follow up in gastroenterology clinic persistently elevated ALP noted. DISCUSSION: Elevated intestinal alkaline phosphatase has been reported in maintaining of gut hemostasis and elevated levels noted in various pathological conditions of the gastrointestinal tract. Meal induced elevation of intestinal ALP has been proposed as a treatment modality to preserve gut hemostasis and minimize inflammation. To the best of our knowledge, our case is an extremely rare presentation of elevated intestinal ALP in an individual with blood group AB. Further molecular and genetic studies in the future may help elaborate functions and therapeutic intervention of intestinal ALP

Teasing apart the different size pools of extracellular enzymatic activity in the ocean

Extracellular enzymatic activity (EEA) is performed by cell-associated and cell-free (i.e., “dissolved”) enzymes. This cell-free fraction is operationally defined as passing through a 0.22 μm filter. The contribution of cell-free to total EEA is comparable to the cell-associated counterpart, so it is critical to understand what controls the relative importance of cell-free versus cell-associated EEA. However, attempts to tease apart the contribution of EEAs in the so-called dissolved fraction (<0.22 μm) in general, and of the nanoparticle size fraction (0.020–0.20 μm) in particular, to the total EEA pool are lacking. Here we performed experiments with Northern and Southern Hemisphere coastal waters to characterize the potential contribution of that nanoparticle fraction to the total EEA fraction of alkaline phosphatase, beta-glucosidase and leucine aminopeptidase. We found a significant contribution (in both hemispheres) of the nanoparticle fraction to the total EEA pool (up to 53%) that differed depending on the enzyme type and location. Collectively, our results indicate that a significant fraction of the so-called “dissolved EEA” is not really dissolved but associated to nanoparticles, colloidal nanogels and/or viruses. Thus, the total marine EEA pool can actually be divided into a cell-associated, undissolved-cell-free (associated to nano-particle of different origins such as viruses and nanogels) and a dissolved-cell-free pools. Our results also imply that the dissolved EEA pool is more complex than thus far anticipated. Future research will be now needed to further characterize the factors controlling the relative importance of these different pools of EEA, which are key in the recycling of organic matter in the ocean.

Toxic effects of copper sulfate and copper nanoparticles on minerals, enzymes, thyroid hormones and protein fractions of plasma and histopathology in common carp Cyprinus carpio

Differences in toxicological effects of dissolved copper and copper nanoparticles were studied in common carp, Cyprinus carpio. The fish were exposed to 0.25mgL−1 copper as copper sulfate (0.25Cu), 0.25mgL−1 copper as copper oxide nanoparticles (0.25NCu) and 25mgL−1 copper as copper oxide nanoparticles (25NCu) over 14days. Plasma biochemical, enzymatic and hormonal characteristics, and liver and kidney histopathology were examined at the end of the experiment. The results showed that both forms of copper had no significant effects on plasma calcium levels, however, significantly increased plasma phosphorous levels, compared to control group (no added copper). Plasma alanine transaminase (ALT) activity increased in 0.25Cu and 25NCu treatments compared to the control and 0.25NCu treatments. Nanoparticle copper exposure significantly decreased plasma alkaline phosphatase (ALP) activity compared to the control and 0.25Cu treatments. Only copper sulfate exposure caused plasma aspartate transaminase (AST) elevation. Both copper forms increased plasma T4 and free T4 (FT4); however, copper sulfate effect was higher than nanoparticle copper. Copper sulfate exposure increased plasma albumin fraction, whereas, 25mgL−1 copper nanoparticle exposure increased plasma α2-globulin fraction compared to the control. Both copper forms damaged the fish liver and kidney, however, copper sulfate caused more severe damages compared to nanoparticle copper. Overall, except for plasma ALP and α2-globulin fraction, dissolved copper seems to be more toxic than nanoparticle copper in common carp.

Bone matrix vesicle-bound alkaline phosphatase for the assessment of peripheral blood admixture to human bone marrow aspirates

PurposePeripheral blood (PB) admixture should be minimized during numerical and functional, as well as cytokinetic analysis of bone marrow (BM) aspirates for research purposes. Therefore, purity assessment of the BM aspirate should be performed in advance. We investigated whether bone matrix vesicle (BMV)-bound bone alkaline phosphatase (ALP) could serve as a marker for the purity of BM aspirates. ResultsTotal ALP activity was significantly higher in BM serum (97 (176–124)U/L, median (range)) compared to PB serum (63 (52–73)U/L, p<0.001). Agarose gel electrophoresis showed a unique bone ALP fraction in BM, which was absent in PB. Native polyacrylamide gel electrophoresis revealed the high molecular weight of this fraction, corresponding with membrane-bound ALP from bone matrix vesicles (BMV), as evidenced by electron microscopy. A serial PB admixture experiment of bone cylinder supernatant samples, rich in BMV-bound ALP, confirmed the sensitivity of this proposed quality assessment method. Furthermore, a BMV ALP fraction of ≥15% is suggested as cut-off value for minimal BM quality. Moreover, the BM purity declines rapidly with larger aspirated BM volumes. ConclusionThe exclusive presence of BMV-bound ALP in BM could serve as a novel marker to assess purity of BM aspirates.

Perinatal undernutrition alters intestinal alkaline phosphatase and its main transcription factors KLF4 and Cdx1 in adult offspring fed a high-fat diet

Nutrient restriction during gestation and/or suckling is associated with an increased risk of developing inflammation, obesity and metabolic diseases in adulthood. However, the underlying mechanisms, including the role of the small intestine, are unclear. We hypothesized that intestinal adaptation to the diet in adulthood is modulated by perinatal nutrition. This hypothesis was tested using a split-plot design experiment with 20 controls and 20 intrauterine growth-retarded (IUGR) rats aged 240 days and randomly assigned to be fed a standard chow or a high-fat (HF) diet for 10 days. Jejunal tissue was collected at necropsy and analyzed for anatomy, digestive enzymes, goblet cells and mRNA levels. Cecal contents and blood serum were analyzed for alkaline phosphatase (AP). IUGR rats failed to adapt to HF by increasing AP activity in jejunal tissue and cecal content as observed in controls. mRNA levels of transcription factors KLF4 and Cdx1 were blunted in jejunal epithelial cell of IUGR rats fed HF. mRNA levels of TNF-α were lower in IUGR rats. They also displayed exacerbated aminopeptidase N response and reduced jejunal goblet cell density. Villus and crypt architecture and epithelial cell proliferation increased with HF in both control and IUGR rats. Serum AP tended to be lower, and serum levamisole inhibition-resistant AP fraction was lower, in IUGR than controls with HF. Serum fatty acids and triglycerides were higher in IUGR rats and higher with HF. In conclusion, the adult intestine adapts to an HF diet differentially depending on early nutrition, jejunal AP and transcription factors being blunted in IUGR individuals fed HF.

PvulI and Xbal polymorphism of estrogen receptor-α and the results of estroprogestagen therapy in girls with functional hypothalamic amenorrhea – preliminary study

IntroductionThe aim of this study was the long-term prospective evaluation of the effects of estroprogestagen (EP) therapy on the bone mineral density (BMD) of girls with functional hypothalamic amenorrhea (FHA) carrying various PvuII and XbaI polymorphisms of ER-α.Material and methodsProspective observation included 84 FHA girls and 50 controls. The FHA patients were subjected to 4-year sequential therapy with 17β estradiol (2 mg from the 2nd to 25th day of the menstrual cycle) and dydrogesterone (10 mg from the 16th to the 25th day). Hormonal parameters, serum concentration of the bone fraction of alkaline phosphatase (BALP), urine concentration of cross-linked n-telopeptide of type I collagen (Ntx) and BMD were determined before and after the treatment.ResultsSix-month treatment resulted in a marked increase in estradiol (p = 0.001), testosterone and prolactin levels (p = 0.01 both) and a significant decrease in BALP and Ntx (p = 0.001 both). Patients with the PP polymorphism had significantly lower baseline BMD compared to carriers of other polymorphic variants of PvuII (p = 0.003). A significant increase in BMD was observed throughout the entire therapy period, with no significant differences in the yearly dynamics of BMD changes observed amongst various polymorphic variants and haplotypes of ER-α.ConclusionsThe EP therapy is effective in the treatment of BMD disorders associated with FHA, and treatment results do not depend on PvuII and XbaI polymorphisms of ER-α.

Bone mineral density in girls with functional hypothalamic amenorrhea subjected to estroprogestagen treatment – a 4-year prospective study

The aim of this study was to evaluate the effects of 4-year estroprogestagen therapy (EP) on the bone mineral density (BMD) of 16- to 17-year-old girls with functional hypothalamic amenorrhea (FHA, n = 78). Baseline values of hormonal parameters, bone fraction of alkaline phosphatase (BALP), and cross-linked n-telopeptide of type I collagen (Ntx) were taken along with BMD measurements. Follow-up measurements of laboratory parameters were performed after 6 months of EP treatment. BMD was measured on a yearly basis. Six-month treatment resulted in a marked increase in estradiol levels and a significant decrease in BALP and Ntx. The relative increase in BMD was highest after the second year of treatment. Based on the dynamics of BMD changes during the first year of treatment, we identified a subgroup with no or insignificant reactions to the treatment. It was characterized by significantly higher baseline BMD and markedly lower baseline Ntx compared to the patients who responded to 1-year therapy well or extremely well. Further follow-up proved, however, that this subgroup did not differ significantly in terms of the long-term prognosis for BMD normalization. In conclusion, this study showed that EP therapy is effective in the treatment of BMD disorders associated with FHA.

Markers of the Hepatic Component of the Metabolic Syndrome as Predictors of Mortality in Renal Transplant Recipients

Cardiovascular disease (CVD) is a leading cause of mortality in renal transplant recipients (RTRs). Metabolic syndrome (MS) is highly prevalent in RTRs. Nonalcoholic fatty liver disease (NAFLD) is considered the hepatic component of MS. We investigated associations of NAFLD markers with MS and mortality. RTRs were investigated between 2001 and 2003. NAFLD markers, alanine aminotransferase (ALT), gamma-glutamyl transferase (GGT) and alkaline phosphatase (AP) were measured. Bone and nonbone fractions of AP were also determined. Death was recorded until August 2007. Six hundred and two RTRs were studied (age 52+/-12 years, 55% men). At baseline 388 RTRs had MS. Prevalence of MS was positively associated with liver enzymes. During follow-up for 5.3[4.5-5.7] years, 95 recipients died (49 cardiovascular). In univariate Cox regression analyses, GGT (HR=1.43[1.21-1.69], p<0.001) and AP (HR=1.34[1.11-1.63], p=0.003) were associated with mortality, whereas ALT was not. Similar associations were found for cardiovascular mortality. Adjustment for potential confounders, including MS, diabetes and traditional risk factors did not materially change these associations. Results for nonbone AP mirrored that for total AP. ALT, GGT and AP are associated with MS. Of these three enzymes, GGT and AP are associated with mortality, independent of MS. These findings suggest that GGT and AP are independently related to mortality in RTRs.

Effect of calcium on rat intestinal alkaline phosphatase activity and molecular aggregation

Two fractions of rat intestinal alkaline phosphatase (IAP) were detected by Western blot: 168 ± 6 and 475 ± 45 kDa. The low molecular weight fraction constitutes 43% of the isolated proteins exhibiting 82% of the enzymatic activity, and a heavier fraction constitutes 57% of the isolated proteins and has 18% of the enzymatic activity. Calcium produced an increase of the 475-kDa form to the detriment of the 168-kDa form. This work also describes the kinetic and structural changes of IAP as a function of calcium concentration. With [Ca2+] < 10 mmole/L, the Ca2+-IAP interaction fitted a binding model with 7.8 ± 4.4 moles of Ca2+ /mole of protein, affinity constant = 19.1 ± 8.4 L/mmole, and enzymatic activity increased as a linear function of [Ca2+] (r = 0.946 p < 0.01). On the other hand, with [Ca2+] >10 mmole/L the data did not fit this model and, the enzymatic activity decreased as a function of [Ca2+] (r = − 0.703 p < 0.05).

Role of heat stable fraction of alkaline phosphatase as an adjunct to ca 125 in monitoring patients of epithelial ovarian carcinoma

Heat stable fraction (HSF) of alkaline phosphatase (ALP) was evaluated as an adjunct to CA 125 as a tumour marker for epithelial ovarian cancer in a follow-up study. In our study group 63.4% of patients had elevated HSF levels (≥10U/L) and 93.3% had elevated CA 125 levels (>35U/mL). The sensitivity of CA 125 and HSF was 93.3% and 63.3% respectively. The decline in the activity of HSF, over the pre-op levels was highly significant after the first (p=0.001) chemotherapy cycle and significant after the second and third cycles (p<0.029). Thereafter, HSF activity was almost undetectable. The decrease in CA 125 levels over the pre-op levels was significant after the first, second, third (p<0.001) and fourth (p<0.034) chemotherapy cycle. HSF can be used alone or as an adjunct to CA 125 in screening and monitoring patients of ovarian carcinoma especially in remote areas where sophisticated facilities are not available and in patients in which CA 125 levels are not raised preoperatively.

Changes in Bone Mineral Density and Selected Metabolic Parameters Over 24 Months Following Renal Transplantation

Our aim was to evaluate changes in serum levels of selected bone metabolism indicators and bone density over 24 months following renal transplant. A partial objective was assessment of the effectiveness of prophylactic administration of vitamin D and calcium preparations to prevent progression of osteopathy after kidney transplantation. Forty patients after kidney transplantation were prophylactically given vitamins A and D (800 IU) and calcium (1000 mg) a day. During monitoring, the serum creatinine in all recipients was <200 μmol/L (subgroup A with creatinine concentration < 120 μmol/L versus subgroup B with creatinine 120 to 200 μmol/L). The concentration of serum parathormone, serum level of bone fraction of alkaline phosphatase, serum concentrations of phosphorus and calcium urinary 24-hour excretion of phosphorus and calcium were examined at 2 weeks and 2 years after transplantation. In the same time period, radiographs of thoracic, lumbar spine, and hip joints were obtained. Bone density (BMD) of the lumbar (L) spine and the hip was determined by dual-energy X ray (Lunar Prodigy). Two years after transplantation in subgroup A, the BMD showed decrease in 80% of recipients in the L spine area but hip showed a 15% BMD increase. In subgroup B, the BMD decreased in 95% recipients in L and hip and only 25% showed a BMD increase. No clinical or radiographic sign of fracture was detected in this group. We conclude that prophylactic administration of vitamin D and calcium is not sufficient to prevent the progression of osteopathy after renal transplantation. Changes in bone density evaluated after the kidney transplantation are affected by graft function.

Second trimester calcium-phosphorus-magnesium homeostasis in women with threatened preterm delivery.

The effect of threatened preterm delivery on calcium, phosphorus, magnesium homeostasis in the second trimester of pregnancy was investigated. Serum concentrations of total and ionized calcium, inorganic phosphorus, magnesium, total protein, albumin, total estrogens and human placental lactogen were determined in women with threatened preterm delivery at 23-28 weeks of gestation (the studied group) and in women with uncomplicated pregnancy of the same duration (the control group). Additionally activities of total alkaline phosphatase and heat-stable alkaline phosphatase fraction were measured. Patients of the studied group compared to the control group showed decreased concentration of total calcium (2.15 +/- 0.073 vs. 2.25 +/- 0.11 mmol/l, P < 0.001), inorganic phosphorus (1.21 +/- 0.26 vs. 1.34 +/- 0.22 mmol/l, P < 0.01) and magnesium (0.63 +/- 0.053 vs. 0.71 +/- 0.12 mmol/l, P < 0.001), total protein (64.0 +/- 5.4 vs. 68.6 + 1.0 g/l, P < 0.001), albumin (546.3 +/- 55.1 vs. 579.6 +/- 49.3 micromol/l, P < 0.003) and placental lactogen (3664 +/- 1806 vs. 4651 +/- 1858 ng/ml, P < 0.02); they also demonstrated decreased activity of total alkaline phosphatase (42.17 +/- 16.99 vs. 50.66 +/- 6.56 IU/l, P < 0.001) and its heat stable fraction (22.65 +/- 7.89 vs. 31.89 +/- 9.09 IU/l, P < 0.001). Patients of the studied group showed normal values of ionized calcium and total estrogens. Premature uterine contractility in women in the second trimester is accompanied by lowered serum concentrations of total calcium, inorganic phosphorus, magnesium, total protein and albumin. There is also decreased activity of total alkaline phosphatase, its heat-stable fraction and placental lactogen which may have diagnostic value. Premature uterine contractility in women in the second trimester may be related to the disturbances of calcium-phosphorus-magnesium homeostasis and calcium supplementation should be considered.

Efficacy and safety of long-term oral falecalcitriol treatment in patients with renal osteodystrophy

We investigated, in a multicenter study, the efficacy and safety of long-term administration of falecalcitriol, a new active vitamin D3, in patients with renal osteodystrophy of the osteitis fibrosa type associated with secondary hyperparathyroidism caused by chronic renal failure. Falecalcitriol was orally administered every day for 48 weeks. Administration was started at a dosage of 0.3 μg/day, and the dosage was changed whenever necessary according to serum calcium (Ca) level. As a result, significant inhibition of the bone resorption markers, i.e., intact parathyroid hormone (i-PTH), pyridinoline (Pyr), and deoxypyridinoline (D-Pyr), was observed from the 8th week, and the bone formation markers, i.e., total activity and bone fraction of alkaline phosphatase, were also significantly inhibited from the 12th week. The bone mineral density (BMD) change rate in the bones of the whole body determined by dual-energy X-ray absorptiometry remained almost constant. When subjects were stratified according to the inhibition rate of bone metabolic parameters, BMD tended to increase in the group with strong inhibition and to decrease in the group with weak inhibition. Mean serum Ca level significantly increased from 9.5 mg/dl, but mean level was subsequently maintained at about 10 mg/dl until the end of administration by adjustment of the doses. These findings suggested that falecalcitriol may inhibit and normalize accelerated bone metabolic turnover without inducing excessive increases in serum Ca level in secondary hyperparathyroidism. With respect to safety, no specific adverse reactions associated with the prolonged administration period were observed.