Leticia G Rao

St Michael's Hospital and University of Toronto, Toronto, Canada

Osteoporosis is a systemic disease that is manifested by a decrease in bone mass and bone microarchitectural deterioration resulting in an increased risk of fracture. Evidence shows that early postmenopausal women lose bone at an accelerated rate. Nutritional recommendations to prevent bone loss include a daily intake of 1500 milligrams of calcium...

Dietary antioxidants play an important role against oxidation, an underlying mechanism in the incidence of chronic diseases. Greens+ is a commercially available preparation containing a variety of plant-derived ingredients. The aim of the current study was to evaluate the antioxidant potential of the methanolic extract of greens+ powder using in vi...

Unlabelled: To date, no intervention studies have been published demonstrating the effect of the antioxidant lycopene on bone. Postmenopausal women supplemented with lycopene had significantly increased antioxidant capacity and decreased oxidative stress and the bone resorption marker N-telopeptide (NTx). Lycopene decreases bone resorption markers...

Background and Objectives: Lycopene is a carotenoid commonly found in tomatoes and tomato products which acts as an antioxidant to decrease oxidative stress and osteoporosis risk. We wanted to determine the effects of a lycopene-restricted diet on oxidative stress parameters and bone turnover markers in postmenopausal women. Setting: St. Michael's...

Lycopene is a carotenoid commonly found in tomatoes and tomato products which acts as an antioxidant to decrease oxidative stress and osteoporosis risk. We wanted to determine the effects of a lycopene-restricted diet on oxidative stress parameters and bone turnover markers in postmenopausal women. St. Michael 's Hospital, Toronto, ON, Canada. PART...

Polymorphisms of the paraoxonase 1 (PON1) enzyme affect the ability to protect LDL from oxidation. Oxidative stress is a risk factor for osteoporosis and antioxidants may be beneficial for prevention. The aim of this study was to determine whether PON1 genotypes modified the association between lycopene and bone turnover markers and oxidative stres...

Pseudohypoparathyroidism (PHP) is characterized by a lack of response to parathyroid hormone (PTH); however, normal skeletal responsiveness to PTH in some patients with PHP type Ia was previously suggested on the basis of clinical observations. To test this hypothesis, we measured cyclic adenosine monophosphate (cAMP) production in response to vari...

A young girl had tibial osteotomies at age 14 for genu valgum and then had recurrent tibial cysts over a number of years. Hypocalcemia and hyperphosphatemia were first noted at age 21. The diagnosis of pseudohypoparathyroidism was made at age 28, when elevated plasma PTH was detected. Clinical and biochemical features, including a PTH response test...

Bone cells derived from human trabecular explants display osteoblastic features. We examined the modulation of alkaline phosphatase activity and cAMP production as the result of exposing trabecular explants to physiologic concentrations of dexamethasone for 4 weeks during cellular outgrowth and subculture. Cells treated with dexamethasone were obse...

Lycopene is an antioxidant associated with a reduced risk of chronic diseases common in women such as osteoporosis and cancer; however, no official recommendation for lycopene consumption exists, and intake data from Canadian women are limited. This study was designed to generate information about average lycopene intake in Canadian women of differ...

We have described and compared culture systems for proximal tubule cell (PTC) preparations from dog and rat kidney. Cells were prepared from kidney cortex by enzyme digestion and purified on Percoll density gradient. The dog PTC and rat PTC differed in their growth characteristics in culture. Although the dog PTC tended to overgrow the contaminatin...

Previous findings in our laboratory have shown that hPTH-(53-84) stimulates alkaline phosphatase activity in dexamethasone-treated ROS 17/2.8 cells. In the present study, we examined the effects of hPTH-(53-84) and hPTH-(1-34) on the expressions of alkaline phosphatase, osteocalcin, and collagen type I mRNA levels in the human osteosarcoma cell lin...

Background and aims: Oxidative stress has been associated with osteoporosis. Greens+™ is a commercially available nutritional supplement containing antioxidative polyphenols. Methods: To study the effects of greens+™ on differentiation and bone formation in human osteoblasts, the cells were cultured in Ham's F-12 medium in the absence or presenc...

Consumption of green tea, a rich source of epigallocatechin-3-gallate (EGCG), is associated with increased bone mineral density. However, the direct effects of this flavonoid on bone mineralization are not known. The present study demonstrates the effects of EGCG on the formation of mineralized bone nodules by SaOS-2 human osteoblast (HOB)-like cel...

Oxidative stress is an important contributor to the risk of chronic diseases. Dietary guidelines recommend increased consumption of fruits and vegetables to combat the incidence of human diseases such as cancer, cardiovascular disease, osteoporosis and diabetes. Fruits and vegetables are good sources of antioxidant phytochemicals that mitigate the...

Mixed isomers of conjugated linoleic acid (CLA) have been shown to have variable effects on bone formation and resorption in animals. The variable effects of CLA on bone physiology may be due to the different isomers present in common commercial preparations of CLA, and the effects of the predominant individual isomers (9cis,11trans and 10trans,12c...

We developed non-invasive, cell-based screening assays to rapidly and biologically assess factors that modulate prostate cancer growth and affect androgen receptor (AR) activity. LNCaP cells, which stably express enhanced green fluorescent protein (EGFP) either constitutively or upon AR activation, were treated with a variety of agents, and then mo...

Many individuals cannot obtain the optimum calcium requirement from food for a variety of reasons. Therefore, calcium supplements are important sources of dietary calcium. One of the calcium sources commercially available is LactoCalcium (milk minerals) that has 28% calcium, and a 2:1 ratio of calcium to phosphorus. The objectives of this study wer...

Oxidative stress induced by reactive oxygen species (ROS) is associated with the risk of osteoporosis, and can be reduced by certain dietary antioxidants. Lycopene is an antioxidant known to decrease the risk of age-related chronic diseases, such as cancer. However, the role of lycopene in osteoporosis has not yet been investigated. In a cross-sect...

Oxidative stress is now recognized as an important etiological factor in the causation of several chronic diseases including cancer, cardiovascular diseases, osteoporosis, and diabetes. Antioxidants play an important role in mitigating the damaging effects of oxidative stress on cells. Lycopene, a carotenoid antioxidant, has received considerable s...

We have shown that osteoblastic cells derived from trabecular bone explants of osteoporotic subjects (OP cells) exhibited an altered alkaline phosphatase (ALP) response to 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] compared to control (CON) cells. Our hypothesis that OP cells have other intrinsic abnormalities was investigated using our cell models rep...

PTH is a major systemic regulator of the concentrations of calcium, phosphate, and active vitamin D metabolites in blood and of cellular activity in bone. Intermittently administered PTH and amino-terminal PTH peptide fragments or analogs also augment bone mass and currently are being introduced into clinical practice as therapies for osteoporosis....

There is evidence to suggest an association between oxidative stress induced by reactive oxygen species (ROS) and chronic diseases. Antioxidants, by virtue of their ability to mitigate the damaging effects of ROS, have generated interest in their use as chemopreventive agents for a number of chronic diseases. Lycopene is the most potent antioxidant...

We studied the of effects raloxifene alone or in combination with human PTH (hPTH) 1-34 in mineralizing cultures of SaOS-2 cells. Raloxifene (10(-8)-10(-6) M) increased bone nodule formation in cultures of SaOS-2 cells when added intermittently from day 8 to day 17. A single 24-h treatment with 10(-8) M hPTH (1-34) at day 8 reduced the nodule area...

Although it is well established that estrogen inhibits bone resorption, its effects on bone formation remain controversial. We studied the effects of intermittent and continuous treatment with estrogen on bone formation in vitro using long term cultures of SaOS-2 cells under conditions that permit mineralization. SaOS-2 cells cultured in dexamethas...

The role of lycopene in human health and disease was studied. The chemistry, bioavailability and metabolism of the carotenoid phytonutrient, which is the most potent antioxidant occurring naturally in plants and vegetables was discussed. Lycopene's bioactivity in prostate cancer, coronary heart disease, and osteoporosis was elucidated.

Osteoclasts have been shown to produce reactive oxygen species (ROS) that can stimulate bone resorption. We explored the hypothesis that lycopene, the antioxidant carotenoid from tomatoes, can inhibit mineral resorption by inhibiting osteoclast formation and the production of ROS. Cells from bone marrow prepared from rat femur were plated into 16-w...

We explored the possibility that lycopene, a carotenoid that is abundant in tomatoes, has effects on proliferation and differentiation of osteoblasts, the cells responsible for bone formation. Human osteoblast-like osteosarcoma SaOS-2 cells were cultured for 24 hours, after which varying doses of a water-dispersible microemulsion preparation of lyc...

Epidemiological studies have shown an inverse relationship between serum lycopene levels and the risk of prostate cancer. The objective of this study was to measure the effect of lycopene on the proliferation of LNCaP human prostate cancer cells in culture. A new, water-dispersible lycopene in an appropriate vehicle was used. The stock solution was...

We compared the effects of 1alpha,25-dihydroxyvitamin D3 [1alpha,25(OH)2D3] and its analog, 1alpha,25-dihydroxy-16-ene-vitamin D3 [1alpha,25(OH)2-16-ene-D3], as well as their interactions with 17-beta estradiol (E2) on osteoblastic function in our human normal (HOB) and osteosarcoma SaOS-2 cell models representing two different stages of differenti...

It is now well established that estrogen inhibits bone resorption. However, its effect on bone formation remains controversial. We studied the effect of 17beta-estradiol (E2) on mineralized bone nodule formation in long-term cultures of osteosarcoma SaOS-2 cells. We showed that SaOS-2 cells formed mineralized nodules which under electron microscopy...

We compared the effects of 1α,25-dihydroxyvitamin D3 [1α,25(OH)2D3] and its analog, 1α,25-dihydroxy-16-ene-vitamin D3 [1α,25(OH)2-16-ene-D3], as well as their interactions with 17-β estradiol (E2) on osteoblastic function in our human normal (HOB) and osteosarcoma SaOS-2 cell models representing two different stages of differentiation, the more dif...

The role of estrogen deficiency in the pathogenesis of postmenopausal osteoporosis is well established. Despite this, the cellular mechanisms of action of estrogen in bone are complex and far from clear. In vitro studies of cellular and molecular biology have concentrated on rodent cells. However, species differences have been noted, and an analysi...

We compared the separate effects of 1alpha,25-dihydroxyvitamin D3 (1alpha,25(OH)2D3) and its analog, 1alpha,25-dihydroxy-16ene,23yne-vitamin D3 (1alpha25(OH)2-16ene,23yne-D3), as well as their interactions with 17-beta estradiol (E2) in our human osteosarcoma SaOS-2 cell models representing two stages of differentiation, the SaOS+DEX and SaOS-DEX c...

We have previously shown that the combination of estrogen (E2) and 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] enhanced alkaline phosphatase (ALP) activity in human osteosarcoma SaOS-2 cells which had been grown in the presence of 10 nmol/L dexamethasone (SaOS + DEX cells). To determine whether this increase in ALP activity was associated with changes i...

We previously developed two models of human osteoblasts with distinct differentiation stages using cells derived from iliac crest trabecular bone explants cultured long term in the presence (HOB + DEX) and absence (HOB - DEX) of 10 nM dexamethasone (DEX) (Wong et al., J Bone Miner Res 1990;5:803). Using these models from 36 subjects aged 41-80 year...

We tested the effect of osteoblastic differentiation on the interactive effects of 17β-oestradiol (E 2 ) and 1,25-dihydroxyvitamin D 3 (1,25(OH) 2 D 3 ) on alkaline phosphatase activity. As cell models we utilized the more differentiated human osteosarcoma (SaOS) cells that had been cultured for 6 days in medium containing 10 n m dexamethasone (Dex...

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We have previously shown that osteoblasts derived from trabecular bone explants and cultured long term in 10 nM dexamethasone ((HOB + DEX) cells) exhibited properties consistent with a more differentiated phenotype compared with those grown in the absence of dexamethasone ((HOB-DEX) cells). To characterize these two cell models further, we measured...

We have examined bone cells derived from iliac crest trabecular explants of 30 patients with idiopathic osteoporosis and 45 control subjects in order to determine whether intrinsic abnormalities in osteoblast function may contribute to the decreased bone formation observed in this disease. Bone cells isolated from all subjects expressed several in...

We studied the effects of 17 beta-estradiol (E2) and PTH on alkaline phosphatase activity in SaOS-2 cells that had been passaged and grown continually in the presence (SaOS + Dex) or absence (SaOS - Dex) of 10(-8) M dexamethasone (Dex). We showed that the more differentiated SaOS + Dex cells had higher alkaline phosphatase activity and PTH-responsi...

We studied the effects of 17 beta-estradiol (E2) and PTH on alkaline phosphatase activity in SaOS-2 cells that had been passaged and grown continually in the presence (SaOS + Dex) or absence (SaOS - Dex) of 10(-8) M dexamethasone (Dex). We showed that the more differentiated SaOS + Dex cells had higher alkaline phosphatase activity and PTH-responsi...

We tested whether the protein kinase C (PKC) modulation of PTH-sensitive adenylate cyclase in ROS 17/2.8 cells is affected by the glucocorticoid dexamethasone and the vitamin D hormone 1,25-dihydroxyvitamin D3 [1,25(OH)2D3]. Basal and PTH- and forskolin-stimulated adenylate cyclase activities were determined in the presence or absence of 100 nM pho...

We tested the effects of various parathyroid hormone (PTH) peptides on alkaline phosphatase (ALP) activity in the osteoblastic cell line ROS 17/2.8. In dexamethasone-treated ROS 17/2.8 cells there was a dose-related increase in ALP activity due to treatment with hPTH (53-84). ALP activity was stimulated by 10 nM hPTH (53-84) by a mean of 1.51 +/- 0...

Both parathyroid hormone (PTH)- and forskolin-stimulated adenylate cyclase activities in ROS 17/2.8 cells are enhanced by increasing the medium concentrations of CaCl2 from 10(-5) M to 3 x 10(-3) M. The ED50 for CaCl2 for both PTH- and forskolin-stimulated activities are similar. The tumor-promoting phorbol ester phorbol 12-myristate 13-acetate (PM...

The calcium modulation of the cyclic 3',5'-adenosine monophosphate (cAMP) response to parathyroid hormone (PTH) was studied in a clonal osteosarcoma cell line ROS 17/2.8. CaCl2 was found to stimulate the PTH-sensitive cAMP response of intact cells. At the maximal concentration of 1 mM CaCl2, the maximum response to PTH was increased, but the ED50 f...

Previous studies in our laboratory have demonstrated relatively large numbers of cell surface binding sites for the carboxylterminal (53-84) region of PTH on ROS 17/2.8 rat osteosarcoma cells, a clonal osteoblast-like cell line. In order to gain insight into the significance of these carboxylterminal binding sites, we studied the effect of intact b...

In short-term experiments, male Wistar rats were made diabetic for 10 days with a single injection of streptozotocin (65 mg/kg body weight). One group of diabetic rats was treated with insulin for 3 days prior to sacrifice. In long-term experiments, vitamin D replete or vitamin D depleted rats were made diabetic for 6 weeks. Criteria for diabetes w...

Most studies of PTH receptor binding have been carried out with amino-terminal radioligands which only detect binding within that region of the hormone molecule. We studied the binding of electrolytically labeled intact bovine PTH [bPTH-(1-84)], and its amino-terminal fragment, bPTH-(1-34) to intact cloned rat osteosarcoma cells (ROS 17/2.8). We al...

We describe the functional characteristics of a new primary culture system derived from a suspension of dog proximal tubular cells. The culture system is maintained on alpha minimal essential medium with 15% fetal calf serum supplementation. At confluency the cultured cells demonstrate the following: (i) typical epithelial morphology using light mi...

Cross-reactivity to some component of rat erythrocytes with an antiserum to parathyroid hormone (anti-PTH) was detected in fixed demineralized sections of bone using the peroxidase-antiperoxidase method. The cross-reactivity was eliminated by the preabsorption of anti-PTH with fixed, washed rat erythrocytes. This technique provides an easy and rapi...

Deparaffinized sections of fixed decalcified neonatal rat radii were incubated in bovine PTH (bPTH; 1-10 MRC units/ml) or in PTH-solvent. They were then stained for PTH by the peroxidase-antiperoxidase method using guinea pig antiserum to bPTH and the substrate 3,3'-diaminobenzidine-H2O2. Staining caused by nonspecific binding of PTH to the bone ma...

Deparaffinized sections of fixed decalcified radii of 2- to 3-day-old rats were incubated in either salmon calcitonin (sCT; 0.1-1.0 MRC U/ml) or in hormone-solvent and then stained for CT by the peroxidase-antiperoxidase method using rabbit antiserum to sCT and 3,3'-diaminobenzidine-H2O2. The sCT-treated radii showed intense brown staining over the...

Specific antibodies to porcine gingival type III collagen were raised in sheep. After purification on collagen affinity columns the antibodies were used for immunohistochemical localization of type III collagen in porcine periodontal and dental tissues employing the peroxidase-antiperoxidase (PAP) procedure. The extent of staining of the periodonta...

Antibody against Type I collagen was raised in rabbits and purified by immunoadsorption on Sepharose-conjugated Types I and III collagen. The cross-reactivity of purified antibody to Type III collagen was found to be less than 0.5% by passive haemagglutination and less than 1.5% by radioimmunoassay. When paraffin sections of fixed and decalcified p...

The cells were evaluated for their responses to the calcium-regulating hormones prostaglandin E1 (PGE1), parathyroid hormone (PTH) and calcitonin (CT) by measuring the concentration of intracellular cyclic-3′,5′-adenosine monophosphate, cAMP. The primary cultures of mixed epithelial cells and fibroblast-like cells responded to 2.5 μg/ml of PGE1 wit...

Five different cell populations, designated I to V, were isolated from minced newborn rat calveria by 5-sequential 20 min incubations with an enzyme mixture containing collagenase, elastase and DNAse. In primary culture, all five populations responded to parathyroid hormone (PTH) to a different degree, population IV giving the highest increase in c...

The amino acid sequence of peptides isolated from a tryptic digest of penicillopepsin (EC 3.4.23.7), a subtilisin (EC 3.4.21.14) digest of maleylated penicillopepsin, and a chymotryptic digest of penicillopepsin modified with dinitrophenylsulfenyl (DNPS) chloride have been determined. The first two digests identified four of the five lysyl residues...

The digest of penicillopepsin (EC 3.4.23.7) with protease II from Myxobacter AL-1 gave five fragments which were separated on a Biogel P-100 column in 70% formic acid. The fragments were from 16 to 125 amino acids long. Two fragments were also isolated from a digest with a protease from Staphylococcus aureus. The analysis of these fragments by auto...

The determination of the amino acid sequences of 70 peptides obtained from a thermolytic digest of penicillopepsin (EC 3.4.23.7) is described. Fifty-six unique sequences ranging from 2 to 13 amino acids were compiled. Among these was a heptapeptide whose sequence is nearly identical with that of the epoxide-reactive active site peptide of porcine p...

The action of penicillocarboxypeptidase-S1 on a number of peptides obtained from enzymatic digests of penicillopepsin and other sources has been studied and compared with that of pancreatic carboxypeptidase A. In addition to proline, lysine, and arginine, penicillocarboxypeptidase-S1 readily cleaved glycine, and glutamic and aspartic acids from pep...

A three-dimensional nine element HF-CW Doppler sounder array has been used to detect ionospheric disturbances which may be due to tornadoes. The typical events chosen in the present study occurred on November 20 and 27, 1973. Both events are apparently associated with tornadoes sighted in the Huntsville, Alabama area. The Doppler records show S-sha...

The coupling between the lower and upper atmosphere during severe weather and thunderstorms is investigated from the ionospheric Doppler frequency fluctuations recorded using a three dimensional CW Doppler sounder array. Two events typical of the severe thunderstorms that occurred on May 26, 1973 and March 20, 1974, are chosen for our investigation...

The reaction of elastase with trinitrobenzene sulfonic acid was investigated in the pH range 9–12. Elastase was found to be inactivated by 2,4,6-trinitrobenzene sulfonic acid. The pH dependence of the pseudo first-order inactivation rate constant showed a pK of 10.3 and gave a Hill plot coefficient of 1.15. Trinitrophenol did not inactivate the enz...