Urinary Hydroxyproline Is Only Suitable As a Biomarker for Acute Intake, Up to 6 hr Postingestion of Collagen Proteins in "Free-Living," Healthy, Active Males.

The urinary excretion of hydroxyproline (Hyp), abundant in collagen protein, may serve as a biomarker of habitual collagen intake, assisting with investigations of current interest in the role of dietary collagen intake in supporting the synthesis of collagenous body tissues. This study investigated the time course of urinary Hyp excretion in "free-living," healthy, active males following the ingestion of a standardized bolus (20 g) of collagenous (gelatin and a hydrolyzed collagen powder) and dairy (calcium caseinate and hydrolyzed casein) proteins. The excretion of Hyp was assessed over a 24-hr period, separated into three collection periods: 0-6, 6-12, and 12-24 hr. Hyp was elevated for 0-6 hr after the consumption of collagen-containing supplements (gelatin 31.3 ± 8.8 mmol/mol and hydrolyzed collagen 33.7 ± 22.0 mmol/mol vs. baseline: gelatin 2.4 ± 1.7 mmol/mol and hydrolyzed collagen 2.8 ± 1.5 mmol/mol; p < .05), but not for the dairy protein supplements (calcium caseinate 3.4 ± 1.7 mmol/mol and hydrolyzed casein 4.0 ± 3.7 mmol/mol; p > .05). Therefore, urinary Hyp reflects an acute intake of collagenous protein, but is not suitable as a biomarker for quantifying habitual collagen intake, provided through regular dietary practices in "free-living," healthy, active males.

METABOLIC, ENZYMATIC AND MINERAL MAINTENANCE OFREPARATIVE OSTEOGENESIS OF FACIAL CRANIUM INJURIES.

Aim - study of marker enzymes, hormonal and carbohydrate-protein indicators of the state of reparative osteogenesis in patients with complicated and uncomplicated course of injuries of facial cranium. The study included 81 patients with injuries of facial cranium, which were divided into 2 groups, depending on the presence of complications. The following enzyme indicators were studied: the level of excretion of hydroxyproline in daily urine; alkaline and acid phosphatase activity; the percentage of bone isoenzymes of alkaline phosphatase. To assess the mineral metabolism, the level of total and ionized calcium and inorganic phosphorus in the blood serum, as well as their excretion in the urine, were determined. To assess the state of metabolism, the concentration of glycosaminoglycans and their fractions in the blood serum were studied. To study the structural and functional state of the bone tissue the densitometry was performed. In patients with complicated course of injuries of facial cranium assosiated with traumatic brain injury there was revealed the increase (р<0,05) of: excretion of phosphorus, uronic acids and oxyproline, while the excretion of calcium was not disturbed (р>0,05), and excretion of magnesium was decreased (р<0,05). It was found out that the level of calcium of blood serum in patients with complicated course is significantly (р<0,05) lower than in the control group and does not depend on the presence of craniocerebral injury (р>0,05). The decrease of the level of ionized calcium content in blood serum can be the confirmation of lower metabolic activity of reparative osteogenesis processes, first of all at the expense of damage of central mechanisms. When studying the content of carbohydrate-protein metabolites by complicated course of injuries of facial cranium, the absolute increase (р<0,05) of concentration of chondroitin-6-sulfates was revealed, and during the analysis of results it was found out that in absolute values, as well as in structural indexes, the specific weight of various fractions changes, that can be the evidence of instability of mechanisms of osteogenesis and of damage of physiological mechanisms of reparative osteogenesis. Densitometric equivalents of forming of complicated course of injuries of facial cranium are the increase of broadband ultrasonic attenuation and the decrease of its spreading speed on the background of low levels of chondroitin-6-sulfates.

Inhibition of cancer cell mitosis by reducing the availability of phosphate.

The addition of phosphate groups is an essential requirement for the proper functioning of cyclin and cyclin dependent kinase which control various stages in the mitotic division of cancer cells. Thus limiting the availability of phosphate is likely to interfere with the metabolism of rapidly growing malignant cells. The human hormone glucagon and the anti metabolite mithramycin reduce serum phosphate by increasing phosphaturia and are both very effective in treating Paget's disease of bone, a precancerous condition. In this disorder large doses of glucagon given intravenously relieve bone pain and cause serum phosphate and alkaline phosphatase as well as urine hydroxyproline to fall, indicating a marked reduction in bone turnover. A constant iv infusion of glucagon was given to each of three patients all of whom had secondary malignant bone deposits. Two of the patients had primary prostate cancer and one had a squamous cell lung tumour. All three patients had relief of bone pain and a fall in serum alkaline phosphatase. Serum acid phosphatase also fell in the two patients with prostate cancer. It is proposed that the marked drop in serum phosphate due to glucagon causes intracellular phosphate to fall. This in turn disrupts the addition and removal of phosphate groups essential for the proper functioning of cyclin and cyclin dependent kinase. These two proteins control the transition from G1 to S (DNA synthesis phase) and G2 to M (mitotic phase) in the dividing cycle of malignant cells. Depriving a tumour of an essential ingredient used in phosphorylation reactions will disrupt its growth. It is also proposed that, by the same mechanism, glucagon induced hypophosphataemia renders malignant cells more sensitive to established chemotherapeutic agents and radiation waves. If this hypothesis proves to be correct, lowering intracellular phosphate may become an useful tool in cancer therapy. However extensive studies are necessary to determine whether mitosis in cancer cells can be advantageously disrupted by glucagon induced hypophosphataemia.

Metabolomics study of renal fibrosis and intervention effects of total aglycone extracts of Scutellaria baicalensis in unilateral ureteral obstruction rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Scutellariae Radix (Scutellaria baicalensis Georgi) is a well-known Traditional Chinese Medicine (TCM) which mainly contains flavonoids. Our previous studies have demonstrated that total aglycone extracts of Scutellaria baicalensis (TAES) can improve kidney disease in rats. AIM OF THE STUDY: To investigate the renal fibrosis (RF) pathogenesis and TAES treatment mechanism in unilateral ureteral obstruction (UUO) rats, using a metabolomics approach based on gas chromatography-mass spectrometry (GC/MS). METHODS: Rats with RF were divided into 6 groups with rats subjected to sham operation as normal control. The effects of TAES on some RF closely related parameters in UUO rats were investigated. A metabolomics method, based on GC/MS, was developed to monitor metabolic alterations in urine. Multivariate data analysis was utilized to identify biomarkers potentially associated with RF and the anti-RF activity of TAES. Ontology-based enrichment analysis by BiNChE and pathway analysis by MetPA aid in the interpretation of difference metabolites. RESULTS: After 10 days of treatment, the parameters of renal function begin returning to normal, and the abnormal high expressions of genes associated with extracellular matrix (ECM) were relived. In the metabolomics study, metabolic perturbations induced by UUO were reversed after treatment and TAES showed a dose-dependent therapy effect on RF, meanwhile, 18 potential biomarkers associated with RF were identified. Enrichment analysis of metabolites shows an over representation of mostly alkane-alpha, omega-diamine and alpha, omega-dicarboxylic acid, and these biomarkers are primarily involved in Glycine, serine and threonine metabolism, Retinol metabolism, Arginine and proline metabolism and Fructose and mannose metabolism. CONCLUSIONS: Our findings indicate that TAES have positive effects on UUO-induced RF in rats, meanwhile, metabolomics method coupled with metabolites enrichment analysis is a useful tool for revealing the pathogenesis of diseases and action mechanism of TCM on the whole body.

Experimental study of the effect on bone metabolism and bone histomorphometry of osteoporosis rats with birdpecking and revolving moxibustion on twelve back-shu points.

The aim of the study was to explore the effect of birdpecking and revolving moxibustion on twelve back shenshu points on the bone metabolism and bone histomorphometry of osteoporosis rats. The 50 female rats of 8 months old that did not pregnant were collected and randomly divided into sham-operation group, model group, moxibustion group, moxibustion and estrogen group, and estrogen group. All the rats, except for the rats in the sham-operation group, received ovarian surgery to establish the models. After 10 days postoperatively (healing), the rats received moxibustion and estrogen therapy. According to the different groups, the rats received rat femur in vivo bone mineral density assessment at 90 days after surgery. After that, the rats were sacrificed, and then the left femoral bones were collected for bone histomorphometry test; blood was taken for bone alkaline phosphatase (BALP) testing, and urine was collected for hydroxyproline testing. The urine hydroxyproline was tested once at 24 h after ovarian surgery. At 24 h after ovarian surgery, the urine hydroxyproline in the ovariectomy group was significantly higher than that in the sham-operation group (P < 0.05), indicating that after ovarian surgery, the collagen broke down which accelerated the process of osteoporosis. After the intervention therapy with moxibustion and estrogen, the BALP, urinary hydroxyproline and femoral bone histomorphometry were comparatively analyzed, and the results showed that the intervention groups were higher than the model group (P < 0.05). But when compared with the sham-operation group, the indexes in the intervention groups were decreased, and the differences were significant (P < 0.05), indicating that intervention could only delay the incidence of osteoporosis. The Chinese traditional measure of "birdpecking and revolving moxibustion on twelve back-shu points" can effectively prevent the recession of bone metabolism of osteoporosis rats, and slow down the degeneration of bone morphology, which can be used to delay the incidence of osteoporosis.

Suppression of NF-κB p65 nuclear translocation and tumor necrosis factor-α by Pongamia pinnata seed extract in adjuvant-induced arthritis.

Pongamia pinnata is a plant known for its therapeutic usage in Indian traditional medicine. Despite the controversy regarding toxic flavonoid and erucic acid content, the seed of this plant is consumed in tribal medicine and its oil is used in Ayurveda to treat psoriasis and arthritis. This study explored the potential anti-arthritic effects of a P. pinnata seed (hexane) extract (PSE) at non-lethal doses in an adjuvant-induced arthritic rat model; possible mechanisms of any observed effects were also explored. After establishing the lethal doses arising from oral exposure to the extract, the material was administered per os daily at two doses (0.3 g/kg/day; 0.5 g/kg/day) to arthritic rats. Other rats received indomethacin or vehicle (control). Treatments were performed for a total of 14 days. One day after the final exposure, the rats were euthanized to permit harvest of various cells, blood, and tissues for analyses. Paw diameter and tissue myeloperoxidase activity in the paws were evaluated as indices for edema and neutrophil infiltration into the tissue. The severity of arthritis in the experimental rats was assessed via measures of urinary hydroxyproline (HP) and glucosamine, and of serum pro-inflammatory TNFα and anti-inflammatory IL-10. The extent of NF-κB p65 nuclear translocation in peritoneal macrophages harvested from naïve rats and then treated in vitro was also assessed. The results indicated that exposure to PSE significantly decreased paw diameter, tissue myeloperoxidase level, and levels of urinary HP and glucosamine, as well as of serum TNFα and IL-10 in adjuvant-injected (arthritic) rats. In vitro PSE treatment also resulted in a marked inhibition of NF-κB p65 nuclear translocation in primary cultures of peritoneal macrophages. Thus, PSE appears to be able to prevent experimental arthritis, in part, by helping to maintain the balance between pro- and anti-inflammatory cytokines and by inhibiting NF-κB activation.

The relationship between fibroblast growth factor 23 and osteoporosis in postmenopausal women.

AIM: A lack of estrogen in postmenopausal women is an important factor causing the development of osteoporosis. Our purpose is to investigate the effects of Fibroblast Growth Factor 23 (FGF-23) on bone mineral metabolism and bone turnover. METHODS: Twenty-eight patients with postmenopausal osteoporosis (PMO), 32 patients with postmenopausal osteopenia and 30 healthy control subjects (postmenopausal non-osteoporosis) were included in this study. In order to assess the bone mineral metabolism; FGF 23, parathyroid hormone, vitamin D, calcium, phosphate, osteocalcin, alkaline phosphatase and hydroxyproline levels were measured. RESULTS: FGF 23 levels were found significantly higher in PMO group compared with postmenopausal osteopenia and control groups (P<0.01 and P<0.05 respectively). Urine hydroxyproline level was detected to be significantly lower in PMO patients compared with control group (P<0.01). Lomber and femur BMD levels were found to be significantly lower in PMO patients compared with postmenopausal osteopenia and control groups (P<0.001, P<0.001; P<0.001, P<0.001 respectively). On the other hand, when we categorized the PMO group subjects according to the age of menopause, the FGF 23 levels were found to be significantly higher in the group of menopausal age <5 years compared to the group of menopausal age >10 and to the group of menopausal age 5-10 years (P<0.05, P<0.05). CONCLUSION: We think our findings indicate that serum FGF 23 level is a significant determinant of increased bone turnover at early periods in PMO patients.

Zinc as a nutritional approach to bone loss prevention in an ovariectomized rat model.

OBJECTIVE: In this study, we have investigated the role of zinc supplementation (a nutritional antioxidant) in an ovariectomized osteopenic rat model. METHODS: Forty-eight female Wistar rats were assigned to four groups: control, zinc, ovariectomy (OVX), and OVX + zinc. Analysis was performed to compare the study groups on bone metabolism markers, bone antioxidant enzymes, and zinc and copper levels in serum and bone tissues. Electron microscopy was also performed to assess morphological changes. RESULTS: Estradiol levels decreased and tartarate-resistant acid phosphatase 5b levels increased in the OVX group. In the OVX + zinc group, these levels were regulated; however, estradiol levels were still significantly lower than those in controls. The OVX group showed significantly higher urinary excretion of hydroxyproline, which recovered upon zinc supplementation but was higher than normal levels. The activities of catalase and superoxide dismutase decreased in ovariectomized animals and up-regulated upon zinc supplementation. Zinc supplementation in the OVX group revoked reduced glutathione levels and elevated malondialdehyde levels. Reduction in zinc and copper levels was observed in the bone tissues and serum of the OVX group. Zinc administration restored these levels to normal. Electron microscopic studies revealed a looser structure and resorbed areas in ovariectomized rat cortical bone. Zinc administration restored bone tissue morphology. CONCLUSIONS: These findings suggest that changes in cortical bone attributed to estrogen deficiency are arrested by zinc supplementation, which can be a sustainable approach to improving bone health.

Antiosteoporotic activity of echinacoside in ovariectomized rats.

PURPOSE: Echinacoside (ECH), isolated from Cistanche tubulosa (Schrenk) R. Wight stems, has been reported to enhance bone regeneration in MC3T3-E1 cells in vitro. The objectives of this study were to investigate the antiosteoporotic effect of ECH on bone metabolism in the ovariectomized (OVX) rat model of osteoporosis in vivo. METHODS: Fifty-six aged 6 months female Sprague-Dawley rats were randomly assigned into sham-operated group (SHAM) and six OVX subgroups (n=8 each). The OVX rats were then subdivided into six groups treated with vehicle (OVX), Xian-ling-gu-bao (XLGB, 0.5 g/kg body weight/day, orally), 17ß-estradiol (E2, 50 µg/kg body weight/day, orally) or ECH (30, 90, and 270 mg/kg body weight, daily, orally) for 12 weeks respectively. We evaluated the pharmacological effects of E2, XLGB and ECH against osteoporosis by evaluating the body weight, uterus wet weight, serum and urine biochemical parameters, bone mineral density (BMD), bone biomechanical properties, bone microarchitecture, bone histomorphology and uterus immunohistochemistry. RESULTS: In OVX rats, the increases of body weight, serum hydroxyproline (HOP) levels, and the decreases of uterus wet weight and BMD were significantly reversed by ECH treatment. Moreover, three dosages of ECH completely corrected the increased urine concentration of calcium (Ca), inorganic phosphorus (P), and HOP observed in OVX rats. Furthermore, Micro-CT analysis results of distal femur showed that all ECH-treated groups notably enhanced bone quality compared to OVX group (p<0.05). Consistent with this finding, total femur BMD and biomechanical strength of tibia were significantly improved (p<0.05) after 12 weeks ECH administration. Histological results also showed the protective activity of ECH through promotion of bone formation and suppression of bone resorption. In addition, the ECH administration also significantly enhanced the expression of ER in the uteri according to immunohistochemical evaluation (p<0.05). Those findings, based on the serum and urine biochemical, BMD, Micro-CT, biomechanical test, histopathological and immunohistochemical parameters, showed that ECH has a notable antiosteoporotic effect, similar to estrogen, especially effective for prevention osteoporosis induced by estrogen deficiency. CONCLUSION: These results suggest that ECH, as a new class of phytoestrogen, has a remarkable antiosteoporotic activity, and may be a promising candidate for treatment of postmenopausal osteoporosis induced by estrogen deficiency in a natural way through herbal resources.

Metabolic regulatory and anti-oxidative effects of modified Bushen Huoxue decoction on experimental rabbit model of osteoarthritis.

OBJECTIVE: To observe the metabolic, regulatory and anti-oxidative effects of modified Bushen Huoxue Decoction (BSHXD), a Chinese herbal medicine for kidney (Shen)-reinforcement and blood-activation, on an osteoarthritis (OA) rabbit model. METHODS: A rabbit model for knee joint OA was established by the classic Hulth's method. The OA model rabbits were randomized into 5 groups: the model control group, the positive control group treated with glucosamine sulfate, and the three BSHXD treated groups treated respectively with low, moderate, and high doses of BSHXD. In addition, a normal control group and a sham-operated group were set up. Experimental animals were sacrificed after a 7-week treatment, and pathological changes in cartilaginous tissue were estimated using the Mankin criteria. Hydroxyproline (Hyp) and malonaldehyde (MDA) contents in blood serum and urine, as well as superoxide dismutase (SOD) activity and nitric oxide (NO) content in blood serum and knee joint synovial homogenates were detected. RESULTS: Mankin scoring showed insignificant statistical differences between the various treatment groups (P >0.05), but all were better than the model control group (P <0.05). Serum and urinary contents of Hyp and MDA as well as serum and synovial levels of NO were significantly lower, but the SOD activity in blood serum and synovial tissue was higher in the BSHXD treated groups than in the model group P <0.01); the effect of BSHXD was dose-dependent to some extent. CONCLUSION: The modified BSHXD shows an effect of improving cartilage metabolism in experimental rabbits with OA, and possesses osteo-chondric protective effects in antagonizing peroxidation injury.

[Biochemical indices of connective tissue metabolism in patients with chronic hepatitis B and C].

37 patients with chronic hepatitis B and C were examined. Patients were divided into 3 groups depending on the degree of connective tissue dysplasia. We investigated: free and protein-bounded hydroxyproline, collagenase activity, total alkaline phosphatase and its bone fraction, creatinine, calcium and phosphorus content in the blood serum and urine. It has been found the dependence of collagen synthesis from the state of connective tissue. The higher is the degree of dysplasia, the more intensive is the process of collagen synthesis (P < 0.05). The index of corellation between protein-bounded and free fraction can be used as a biochemical marker for determination the stage of pathological process in the liver and for monitoring the effectiveness of therapy.

A high molecular weight protein extract of Mastobranchus indicus (Mi-64) having antiarthritic activity in experimental animals.

Mi-64, a high molecular weight protein (130 kDa), obtained from the tissue homogenate of marine polychaete (Mastobranchus indicus) collected from the Indian Sunderban has antiarthritic activity in experimental animals. The FCA-induced arthritis model was developed in Wistar albino rats to evaluate the antiarthritic effects of Mi-64. After FCA induction, the rats were treated with Mi-64 (0.25 and 0.5 mg kg(-1) body weight) for 10 days. We have determined the paw/ankle swellings, urinary hydroxyproline and glucosamine, serum acid and alkaline phosphatases to assess the antiarthritic activity. The levels of interleukin-1 beta (IL-1ß), IL-6, cytokine-induced neutrophil chemoattractant-1 (CINC-1), tumor necrosis factor-alpha (TNF-α), and IL-10 were measured by ELISA. Results showed that Mi-64 significantly reduced paw/ankle swellings and restored the urinary hydroxyproline/glucosamine and serum phosphatases. Mi-64 significantly inhibited the overproduction of IL-1ß, IL-6, CINC-1, and TNF-α and augmented IL-10 production. The data suggest that Mi-64 produced significant antiarthritic effects that may be mediated by balancing the pro- and anti-inflammatory cytokines.

Effect of milk and calcium supplementation on bone density and bone turnover in pregnant Chinese women: a randomized controlled trail.

OBJECTIVE: Calcium demand is increased during pregnancy. However, few randomized controlled trials examined the effects of calcium supplementation on bone mass during pregnancy. This study determined effects of calcium and milk supplementation on maternal bone mineral density (BMD) and bone turnover in pregnant Chinese women with habitual low calcium intake. METHODS: In this randomized controlled trial, 36 Chinese pregnant women (24-31 years, 18 gestational weeks) were randomly assigned to the following three arms (12 each): I, usual diet; II, "I" + 45 g milk powder (containing 350 mg calcium); or III, "II" + 600 mg calcium/day from gestational age of 20 weeks to 6 weeks post-partum (PP). BMD was measured post-treatment using dual-energy X-ray absorptiometry. Dietary intakes, 24-h urinary calcium, bone resorption (urinary hydroxyproline) and formation (serum osteocalcin) biomarkers were examined at the gestational age of 20 and 34 weeks, and 6 weeks PP. RESULTS: A dose-dependent relationship was observed between calcium intake and BMDs. The BMD values were significantly higher in subjects with calcium and milk supplementation than those in the controls at the whole body and spine (p < 0.05) but not at the hip sites. We found significant decreases in changes of urinary hydroxyproline, and significant increases in serum osteocalcin during the intervention period in the calcium/milk intervention groups than those in the control group (all p < 0.05). CONCLUSION: Calcium/milk supplementation during pregnancy is associated with greater BMD at the spine and whole body and suppresses bone resorption in Chinese women with habitual low calcium intake.

Anti-arthritic activity of Indian monocellate cobra (Naja kaouthia) venom on adjuvant induced arthritis.

The Indian Monocellate Cobra venom (NKV) showed anti-arthritic activity over FCA induced arthritis in male albino rats. NKV treatment (1/20th & 1/10th MLD doses x 13 days, i.p.) showed significant restoration in paw & ankle volume, paw weight. Urinary hydroxyproline, glucosamine, serum ACP, ALP and IL-10 level were restored significantly, due to NKV treatment, as compared with arthritic rats. NKV also showed significant protection against arthritis induced oxidative damages. Thus this study confirmed the scientific validation behind ancient belief and use of snake venom in arthritis as mentioned in Ayurveda.

Protective action of aqueous black tea (Camellia sinensis) extract (BTE) against ovariectomy-induced oxidative stress of mononuclear cells and its associated progression of bone loss.

The protective action of aqueous black tea extract (BTE) against ovariectomy-induced oxidative stress of mononuclear cells and its associated progression of bone loss was demonstrated in this study. Eighteen female adult 6-month-old Wistar albino rats were divided into three groups: sham-control (A), bilaterally ovariectomized (B) and bilaterally ovariectomized + BTE supplemented (C). Studies included the measurement of oxidative (nitric oxide, lipid peroxidation) and antioxidative (superoxide dismutase, catalase) markers, inflammatory cytokines (IL-6, TNF-alpha), osteoclast differentiation factor (RANKL) and bone resorption markers (tartrate-resistant acid phosphatase and hydroxyproline). Also quantitative histomorphometry and histological studies were undertaken. The bone breaking force was measured. The results indicate that BTE was effective in preserving and restoring skeletal health by reducing the number of active osteoclasts. Such changes with BTE supplementation were steadily linked with the reduced oxidative stress of mononuclear cells, serum levels of bone resorbing cytokines, osteoclast differentiation factor and resorption markers. The results of the bone breaking force, histological and histomorphometric analyses further supported the hypothesis. This study suggests that BTE has both protective and restorative actions against ovariectomy-induced mononuclear cell oxidative stress and associated bone loss.

Effect of isoflavone-enriched milk on bone mass in ovariectomized rats.

This study examined the effect of isoflavone-enriched milk on bone loss in ovariectomized (OVX) rats. Thirty 6-week-old Sprague-Dawley female rats were divided into two groups: sham-operated and OVX. The OVX group was subdivided into three dietary groups (OVX, non-isoflavone-enriched milk; OVX+Iso, isoflavone-enriched milk; and OVX+Iso+Vit+Ca, isoflavone-, vitamins D and K-, and Ca-enriched mik). After 19 weeks of feeding, the food efficiency ratio and body weight gain in the sham-operated group were significantly lower compared with those in the other groups. The bone alkaline phosphatase and total alkaline phosphatase activities were significantly higher in isoflavone-enriched groups (OVX+Iso and OVX+Iso+Vit+Ca) when compared with the sham-operated group. Urinary excretions of deoxypyridinoline and hydroxyproline were significantly higher with ovariectomy, but mostly normalized in the OVX+Iso and OVX+Iso+Vit+Ca groups. The rats in the OVX+Iso and OVX+Iso+Vit+Ca groups showed higher femur and tibia weights. A significant increase was found in bone density of femur and trabecular bone area in the OVX+Iso+Vit+Ca group, which almost reached that of the sham-operated group, whereas no difference was found among the OVX and OVX+Iso groups. The histological areas of the proximal tibia sections showed highly filled trabecular bone in both isoflavone-enriched groups (OVX+Iso and OVX+Iso+Vit+Ca). The present study indicated that isoflavone-enriched milk may have a partial preventive effect on ovariectomy-induced bone loss; however, vitamins D and K and Ca enrichment with isoflavone may enhance effectiveness for increasing bone mass in OVX rats.

Prolyl-hydroxyproline dipeptide in non-hydrolyzed morning urine and its value in postmenopausal osteoporosis.

BACKGROUND: Owing to the high correlation between the level of prolyl-4-hydroxyproline dipeptide in non-hydrolyzed urine and that of 4-hydroxyproline in hydrolyzed urine, we examined whether the dipeptide might function as a valuable marker of bone turnover. METHODS: Based on densitometric measurements, 68 postmenopausal women were divided into groups of non-osteopathic, osteopenic and osteoporotic subjects. The dipeptide and current urinary resorption markers were assayed in morning urine, the former using liquid chromatography, the others plus serum formation markers by means of immunoassay procedures. Together with the assay of basal levels, diet-related changes and healing effect of yearly alendronate therapy were assessed. RESULTS: Concentration levels in controls and osteoporotic subjects differed significantly; receiver operating characteristics yielded sensitivity of 0.743, specificity of 0.908, area under curve of 0.903, and cut-off of 10.2 micromol/mmol of creatinine. Spearman rank correlation showed the highest pair coefficient between the dipeptide and osteocalcin. Diet-related changes were not found. Following therapy, a significant decline occurred already within a trimester, whilst with the other resorption markers not until 6 months. CONCLUSIONS: The ease of the dipeptide assay in non-hydrolyzed urine surpasses that of hydroxyproline, and the results present the compound as a real competition to other commonly assessed markers in osteopathies.

Comparação dos efeitos do TENS e dos exercícios terapêuticos sobre os níveis de hidroxiprolina na urina em síndrome do impacto no ombro / Comparison of the effects of TENS and exercises therapy effects on hydroxyproline levels in urine on shoulder pain

Objetivo: Comparar os efeitos da neuroestimulação elétrica transcutânea (TENS) e cinesiologia aplicada, assim como esta isolada, na excreção urinária em indivíduos com a síndrome do impacto do ombro (SIO). Métodos: Participaram do estudo dois grupos de 35 indivíduos cada, sendo 30 mulheres e 40 homens, com idade entre 45 e 60 anos. O grupo controle realizou a cinesiologia aplicada e o grupo experimental realizou o tratamento TENS associado à cinesiologia aplicada. Para a mensuração da hidroxiprolina na urina foi utilizado o protocolo de colorimetria. A coleta urinária foi feita na 1ª, 5ª e 10ª sessão. O tratamento foi realizado em 10 sessões de 55 minutos. O tratamento estatístico utilizado foi feito através da análise de variância One Way (ANOVA). Resultado: Não houve melhora significativa como indicado por F = 0,662, p > 0,05. Conclusão: Os resultados mostraram não haver interação significativa entre os tipos de tratamento com a excreção urinária de hidroxiprolina. Contudo, os resultados obtidos das variáveis mostraram uma forte tendência à melhora, apresentando um resultado mais efetivo no grupo que utilizou somente a cinesiologia aplicada até a quinta sessão e, posteriormente, com uma tendência mais efetiva no grupo que utilizou a cinesiologia aplicada + TENS. O estudo mostrou, também, um resultado mais eficiente do grupo que utilizou apenas a cinesiologia aplicada como tratamento.

Objective: To compare the transcutaneous electrical nerve stimulator (TENS) effects associated to the kinesiology applied, and only the kinesiology applied on the hydroxiproline (HP) excretion on individuals with shoulder pain, during 10 physical therapy sessions with duration of 55 minutes each session for both treatments. Methods: The individuals were divided into two groups of 35 people each, being 30 women and 40 men; aged between 40 and 65 years old. The control group underwent only applied kinesiology and the experimental group applied kinesiology associated to TENS. It was used the colorimetric protocol to measure urinary excretion of HP. Three samples of each variable were carried out on the first, fifth and tenth sessions. The ANOVA test with repeated measures to analyze the HP was used for the statistics. Results: There were no significance as indicated by F = 0.662, p > 0.05. Conclusion: We concluded that the study showed a strong benefit tendency for both groups due to HP decrease levels. As a better result before the 5th session for the group applied kinesiology and after the 5th - 10th session of treatment, for the applied kinesiology + TENS group, although there was no significance based on the statistics. And, also, it showed a better result for the group who practiced only applied kinesiology

Effect of nano-calcium-enriched milk on calcium metabolism in ovariectomized rats.

This study was designed to examine the effect of different kinds of calcium enrichment on serum and urine indices of mineral status in ovariectomized rats. Twenty-four 7-week-old Sprague-Dawley female rats were divided into four groups, ovariectomized, and fed diets containing the following: (1) Control, non-Ca-enriched milk; (2) OVX1, calcium carbonate-enriched milk; (3) OVX2, ionized Ca-enriched milk; and (4) OVX3, nano-Ca-enriched milk. After 18 weeks of feeding, the food efficiency ratio in the nano-Ca-fed group was significantly lower compared with those in the Control and OVX2 groups. There was no difference in serum and fecal Ca among the groups. The bone/total alkaline phosphatase ratio was significantly higher in rats fed milk enriched with nano-Ca (59%) and calcium carbonate (62%) than in control (44%) animals. Urinary Ca was the highest in the nano-Ca-enriched group; however, urinary excretions of deoxypyridinoline and hydroxyproline were significantly decreased in the nano-Ca-enriched group. The present results indicate that consumption of nano-Ca-enriched milk resulted in an increase of urinary excretion of calcium and a decrease in deoxypyridinoline and hydroxyproline in ovariectomized rats.

A comparative study on several models of experimental renal calcium oxalate stones formation in rats.

In order to compare the effects of several experimental renal calcium oxalate stones formation models in rats and to find a simple and convenient model with significant effect of calcium oxalate crystals deposition in the kidney, several rat models of renal calcium oxalate stones formation were induced by some crystal-inducing drugs (CID) including ethylene glycol (EG), ammonium chloride (AC), vitamin D(3)[1alpha(OH)VitD(3), alfacalcidol], calcium gluconate, ammonium oxalate, gentamicin sulfate, L-hydroxyproline. The rats were fed with drugs given singly or unitedly. At the end of experiment, 24-h urines were collected and the serum creatinine (Cr), blood urea nitrogen (BUN), the extents of calcium oxalate crystal deposition in the renal tissue, urinary calcium and oxalate excretion were measured. The serum Cr levels in the stone-forming groups were significantly higher than those in the control group except for the group EG+L-hydroxyproline, group calcium gluconate and group oxalate. Blood BUN concentration was significantly higher in rats fed with CID than that in control group except for group EG+L-hydroxyproline and group ammonium oxalate plus calcium gluconate. In the group of rats administered with EG plus Vitamin D(3), the deposition of calcium oxalate crystal in the renal tissue and urinary calcium excretion were significantly greater than other model groups. The effect of the model induced by EG plus AC was similar to that in the group induced by EG plus Vitamin D(3). EG plus Vitamin D(3) or EG plus AC could stably and significantly induced the rat model of renal calcium oxalate stones formation.