Maela Lupo

Physical exercise ameliorates the toxic effect of fluoride on the insulin–glucose system

Daily intake of water with fluoride concentrations >1.5 mg/l produces insulin resistance (IR). On the other hand, physical activity increases insulin sensitivity in the muscle. Therefore, the aim of this study was to evaluate the effect of physical activity on IR in rats treated with sodium fluoride (NaF) in drinking water. Sprague-Dawley rats were divided into three groups (n=10/group): Control (drinking water without NaF), NaF (drinking water with NaF 15 mg/l for 30 days), and Exercise (daily running on a treadmill for 60 min at 2.25 m/min and drinking water with NaF 15 mg/l for 30 days). IR was evaluated with the homeostasis model assessment-IR (HOMA-IR) index using fasting plasma levels of glucose and insulin. IR increased in rats treated with 15 mg/l NaF in drinking water. A decrease in IR was observed in rats that performed physical activity and drank water with 15 mg/l NaF; the Exercise group also showed an increase in the amounts of bone fluoride. The variation in the HOMA-IR values could be the consequence of variation in the sensitivity of tissues to insulin or decrease in plasma fluoride levels due to bone fluoride intake. These findings indicate that the performance of daily physical activity could reduce the negative effects of the chronic ingestion of NaF on glucose homeostasis.

Mathematical model of glucose-insulin homeostasis in healthy rats

According to the World Health Organization there are over 220 million people in the world with diabetes and 3.4 million people died in 2004 as a consequence of this pathology. Development of an artificial pancreas would allow to restore control of blood glucose by coupling an infusion pump to a continuous glucose sensor in the blood. The design of such a device requires the development and application of mathematical models which represent the gluco-regulatory system. Models developed by other research groups describe very well the gluco-regulatory system but have a large number of mathematical equations and require complex methodologies for the estimation of its parameters. In this work we propose a mathematical model to study the homeostasis of glucose and insulin in healthy rats. The proposed model consists of three differential equations and 8 parameters that describe the variation of: blood glucose concentration, blood insulin concentration and amount of glucose in the intestine. All parameters were obtained by setting functions to the values of glucose and insulin in blood obtained after oral glucose administration. In vivo and in silico validations were performed. Additionally, a qualitative analysis has been done to verify the aforementioned model. We have shown that this model has a single, biologically consistent equilibrium point. This model is a first step in the development of a mathematical model for the type I diabetic rat.

Regulation of intestinal calcium absorption by luminal calcium content: Role of intestinal alkaline phosphatase

SCOPE Intestinal alkaline phosphatase is a brush border enzyme that is stimulated by calcium. Inhibition of intestinal alkaline phosphatase increases intestinal calcium absorption. We hypothesized that intestinal alkaline phosphatase acts as a minute-to-minute regulatory mechanism of calcium entry. The aim of this study was to evaluate the mechanism by which intestinal luminal calcium controls intestinal calcium absorption. METHODS AND RESULTS We performed kinetic studies with purified intestinal alkaline phosphatase and everted duodenal sacs and showed that intestinal alkaline phosphatase modifies the luminal pH as a function of enzyme concentration and calcium luminal content. A decrease in pH occurred simultaneously with a decrease in calcium absorption. The inhibition of intestinal alkaline phosphatase by l-phenylalanine caused an increase in calcium absorption. This effect was also confirmed in calcium uptake experiments with isolated duodenal cells. CONCLUSION Changes in luminal pH arising from intestinal alkaline phosphatase activity induced by luminal calcium concentration modulate intestinal calcium absorption.

Computational Model for Studying the Analogies Between Endocrine Systems of Humans and Rats with Diabetes Mellitus. Experimental and Simulated Results.

Abstract As a common laboratory practice, rats are studied as biological models for understanding human endocrine behavior. However, up to now there is not a computational model of these mammals because adjusting a physiological model of the endocrine system is not an easy task. Thus, in this work the first mathematical model of the interaction between insulin and glucose in diabetic rats available to the scientific community is presented. It is based on the diabetic patient model accepted by the FDA in 2008. However, it has a large number of parameters, being most of them very difficult to determine in real patients. The main idea is to obtain a proper in silico diabetic rat which would be able to compare well with the experimental data. It will provide interesting insight about the analogies with humans and, in future, analyze the possibility of doing a realistic scale-up between both, humans and rats. Therefore, the list of model equations and the corresponding parameters, obtained by specific experiments, are described and tested with the simulated results.

Comparison of fluoride effects on germination and growth of Zea mays, Glycine max and Sorghum vulgare.

BACKGROUND Fluorosis is a disease caused by over-exposure to fluoride (F). Argentinas rural lands have higher fluorine content than urban lands. Evidence confirms that plants grown in fluoridated areas could have higher F content. We compared F uptake and growth of crops grown in different F concentrations. The effect of 0-8 ppm F concentrations on maize, soybeans and sorghum germination and growth was compared. After 6 days seeding, the germination was determined, the roots and aerial parts lengths were measured, and vigor index was calculated. F content was measured in each part of the plants. Controls with equal concentrations of NaCl were carried out. RESULTS Significant decrease in roots and aerial parts lengths, and in vigor index of maize and soybeans plants was observed with F concentrations greater than 2 ppm. This was not observed in sorghum seedlings. Also, the amount of F in all crops augmented as F increases, being higher in roots and ungerminated seeds. Sorghum was the crop with the highest F content. CONCLUSION Fluoride decreased the germination and growth of maize and soybeans and therefore could influence on their production. Conversely, sorghum seems to be resistant to the action of F.

In vivo measurement of the rate constant of liver handling of glucose and glucose uptake by insulin-dependent tissues, using a mathematical model for glucose homeostasis in diabetic rats

Diabetes mellitus is a disease that affects glucose homeostasis. The World Health Organization informs that there are over 347 million people in the world with diabetes. The diagnosis and characterization of glucose homeostasis in different metabolic conditions are subjects of great importance with high clinical impact. There are many mathematical models that describe the glucoregulatory system in detail. However, the use of these models is limited because they have a large number of mathematical equations and parameters and they require complex methodologies to estimate of them. This forced to work with average values that decrease the validity of results and the applicability of the models. In this study two mathematical models for rats with diabetes mellitus were developed. The difference between these models and others lies in the possibility of obtaining all parameters for each animal from simple measurements (glucose and insulin plasma levels). Moreover, the models allow to measure in vivo the different physiological processes involved in glucose homeostasis in animals: insulin secretion and its plasma clearance, absorption of insulin from a subcutaneous injection, the liver handling of glucose, intestine absorption of glucose, glucose uptake rate of insulin-independent tissues, glucose uptake rate of insulin-dependent tissues, and renal glucose excretion.

CHAPTER 6:Blood, Plasma and Bone Fluoride Measurement

Fluorine may be found in biological samples in different chemical forms: fluoride: the ionic specie, nonfluoride compound: chemical species that can be converted into fluoride by breaking the bonds that bind to other structures and covalently bound fluorine: compounds where the fluorine is covalently bonded to carbon chains. After consuming fluoridated water, toothpaste with sodium fluoride or sodium fluoride tablets, fluoride is found predominantly in plasma. The concentration of fluoride depends mainly on the dose, time of consumption, renal function and bone remodeling. When monofluorophosphate is administered, nonfluoride compound and fluoride are found in plasma due to the binding of monofluorophosphate to plasma proteins. Fluorine is found predominantly as fluorapatite in bone tissue, and the amount of fluoride in bone is known as the bone fluoride content, which is directly related to the dose, time of exposure, bone formation and physical activity.

Methodology developed for the simultaneous measurement of bone formation and bone resorption in rats based on the pharmacokinetics of fluoride

This paper describes a novel methodology for the simultaneous estimation of bone formation (BF) and resorption (BR) in rats using fluoride as a nonradioactive bone-seeker ion. The pharmacokinetics of flouride have been extensively studied in rats; its constants have all been characterized. This knowledge was the cornerstone for the underlying mathematical model that we used to measure bone fluoride uptake and elimination rate after a dose of fluoride. Bone resorption and formation were estimated by bone fluoride uptake and elimination rate, respectively. ROC analysis showed that sensitivity, specificity and area under the ROC curve were not different from deoxypiridinoline and bone alkaline phosphatase, well-known bone markers. Sprague–Dawley rats with modified bone remodelling (ovariectomy, hyper, and hypocalcic diet, antiresorptive treatment) were used to validate the values obtained with this methodology. The results of BF and BR obtained with this technique were as expected for each biological model. Although the method should be performed under general anesthesia, it has several advantages: simultaneous measurement of BR and BF, low cost, and the use of compounds with no expiration date.

Sequential treatment with monofluorophosphate and zoledronic acid in osteoporotic rats

Abstract Objective Osteoporosis is the consequence of an imbalance in bone remodeling caused by excessive resorption or inappropriate bone formation. This paper proposes a sequential treatment with monofluorophosphate (MFP) and zoledronic acid (Z), together with changes in the calcium content in the diet. Method Seven-week-old female Sprague Dawley rats were divided into five groups (n = 21 per group): (1) sham-operated rats (Sham); (2) ovariectomized (OVX) rats fed with a normal calcium diet (OVX); (3) OVX rats fed with a normal calcium diet and treated sequentially with monofluorophosphate and zoledronic acid (OVX.G1); (4) OVX rats sequentially fed with a low calcium diet and then a high calcium diet, without treatment (OVX.G2); (5): OVX rats fed with a low calcium diet and then a high calcium diet, treated sequentially with monofluorophosphate and zoledronic acid (OVX.G3). Results After 150 days, the OVX.G3 group showed a similar bone volume to that of the Sham group due to an increase in trabecular number. Dual X-ray absorptiometry bone analysis showed an increase of 9.8% compared with OVX rats. Additionally, an increase in the fracture load at the cortical bone and higher fracture load, ultimate load and stiffness in the compression test were found. Conclusion The sequential treatment with monofluorophosphate and zoledronic acid increases trabecular bone mass, bone mineral density and bone strength.

Mathematical model for the homeostasis of alpha-macroglobulins in the rat.

Alpha-macroglobulins (AM) are proteins that inactivate proteinases. Sodium monofluorophosphate (MFP) binds to AM and transiently changes AM plasma levels. As a consequence MFP is useful to modify AM homeostasis. A mathematical model to study the homeostasis of AM is proposed in this paper. The model describes changes in plasma concentration of AM, MFP concentration in the gastrointestinal tract, MFP plasma concentration, plasma concentration of AMMFP and includes rate constants of the processes involved in AM homeostasis. Estimation of the rate constants values was achieved using experimental and mathematical resources. The homeostasis of AM after an oral dose of 80 μmol of MFP was analyzed with a simulation tool. Experimental conditions that modify the homeostasis of AM had been simulated and validated using specific drugs that change some parameter of the system. The mathematical model describes accurately the behavior of the biological model. The results allow concluding that the simplifications made did not underestimate the main processes involved in the homeostasis and, also that the assumptions made were correct.