Coleta de células progenitoras periféricas por aférese automatizada em equino: relato de procedimento / Viability of automated collection of peripheral blood progenitor cells in a horse: report of procedure

The biotechnology used in tendon, bone and joint recoveries in Equine Medicine has improved in recent years. The most used are platelet rich plasma and stem cells from adipose tissue and bone marrow. However, recent studies have shown that stem cells can be found in the bloodstream, also named peripheral blood progenitor cells (CPP). This note aims at reporting the feasibility of automated collection of CPP in horses. The procedure was conducted in an equine, female, Quarter Horses, 2 years old, 385kg. The automated collection of CPP was conducted using apheresis equipment Fresenius- Kabi coupled to C4Y kit. The procedure lasted two hours and 30 minutes without complications, processing 5054mL of whole blood and obtaining 351mL of CPP. Upon completion of the collection, the content of CPP was separated into 10 ml aliquots and immediately stored at -18°C. The automated technique proved to be feasible for horses, but needs improvement in order to achieve greater efficiency and reduce procedure time.(AU)

Padronização da técnica de plasmaférese automatizada em equinos / Standardization of automated plasmapheresis in horses

O presente artigo objetivou estudar a viabilidade da técnica de plasmaférese automatizada e padronizá-la em cinco equinos hígidos, bem como apresentar as complicações durante o procedimento, as adequações em relação aos procedimentos em humanos e avaliar a recuperação de volume globular e proteínas plasmáticas totais nos doadores. Os procedimentos foram realizados com o equipamento Fresenius AS104, com duração média de 1h46min, processamento de 5758mL de sangue total e colheita média de 3133mL de plasma. Não foram observadas alterações significativas do volume globular após a plasmaférese automatizada. A recuperação dos níveis plasmáticos de proteínas foi de 91,4% em 96 horas após o procedimento. A plasmaférese automatizada apresentou-se viável para a espécie equina, diminuindo o tempo de recuperação hematimétrica nos doadores.

This paper aimed to study feasibility and standardize the automated plasmapheresis in five healthy horses, showing the complications during the procedure, adjustments in relation to the procedures in humans and assessing the recovery of globular volume and plasma total proteins in donors. The procedures were performed with the Fresenius AS104 equipment, with an average duration of one hour and forty six minutes, processing 5758mL of whole blood and harvest average of 3133mL of plasma. There were no significant variations in globular volume after the automated plasmapheresis. The recovery of plasma total proteins was 91.4% at 96 hours after the procedure. The automated plasmapheresis appeared viable for the equine species, decreasing the time of hematimetric level recovery in donors.

Effects of depot medroxyprogesterone acetate on the calcium metabolism of adult ovariectomized rats.

This paper describes experiments designed to test the effect of depot medroxyprogesterone acetate (DMPA) on calcium metabolism of adult ovariectomized rats. The 24 animals were randomly assigned to control or treated groups. Treated rats received 15 mg of DMPA i.m. per week, during four or twelve weeks. Controls received solvent alone. The variables characterizing the metabolism of Ca (daily rates of intestinal absorption and excretion, bone accretion and resorption and the sizes of the exchangeable pools and their rate constants) were measured with the aid of 45Ca according to Aubert and Milhaud. No effects were observed at four weeks of treatment. After twelve weeks, treatment produced serum levels of 46.5 +/- 5.6 nmoles of medroxyprogesterone/L, reduction of bone turnover (Ca accretion and resorption rates) and of the size of the slow exchanging Ca compartment. The increase in true Ca intestinal absorption was compensated by the increased endogenous fecal Ca excretion. The mass of body Ca was not affected by treatment.

Effects of depot medroxyprogesterone acetate on the calcium metabolism of adult ovariectomized rats.

This paper describes experiments designed to test the effect of depot medroxyprogesterone acetate (DMPA) on calcium metabolism of adult ovariectomized rats. The 24 animals were randomly assigned to control or treated groups. Treated rats received 15 mg of DMPA i.m. per week, during four or twelve weeks. Controls received solvent alone. The variables characterizing the metabolism of Ca (daily rates of intestinal absorption and excretion, bone accretion and resorption and the sizes of the exchangeable pools and their rate constants) were measured with the aid of 45Ca according to Aubert and Milhaud. No effects were observed at four weeks of treatment. After twelve weeks, treatment produced serum levels of 46.5 +/- 5.6 nmoles of medroxyprogesterone/L, reduction of bone turnover (Ca accretion and resorption rates) and of the size of the slow exchanging Ca compartment. The increase in true Ca intestinal absorption was compensated by the increased endogenous fecal Ca excretion. The mass of body Ca was not affected by treatment.

Hypercalciuria and urinary saturation measurements in climacteric women.

Fifteen percent (20/130) of a group of climacteric women on diets of their choice had urinary calcium (Ca) levels exceeding 4 mg/kg per day. Most of these hypercalciuric subjects had a daily Ca intake of 0.4-0.5 g. Their bone turnover rates were raised and high Ca absorption was observed in 4 cases. Serum Ca and total protein and glomerular filtration rates were normal in all the hypercalciuric patients. The calcium/creatinine (Ca/Cr) ratio (mg l-1/mg l-1, fasting, 09:00-10:00 h) was measured in 72 climacteric women, 35 of whom (49%) had ratios > 0.1. The latter defines a relative hypercalciuria as compared with premenopausal Ca excretion levels. Only 5 of the 35 subjects had calciuria levels above 4 mg/kg per day. The Ca/Cr ratio cannot replace daily urinary Ca measurements for the screening of subjects in whom calciuria may exceed net Ca absorption. Urinary saturation measurements were carried out in 70 women. Supersaturation was observed only in the case of Ca oxalate (CaOx) among several calcium salts usually found in urinary stones. CaOx supersaturation was observed in 95% of the hypercalciuric subjects and in 48% of the rest of the women investigated. The relatively high frequency of CaOx supersaturation can be attributed in part to the decreased excretion of citrate associated with ovarian failure. Oestrogen replacement therapy increased citrate excretion and lowered the level of CaOx supersaturation. Ca supplementation (1 g Ca/day) reduced the degree of supersaturation as a result of the concurrent reduction in oxalate excretion.

Inhibitory effect of fluoride on the secretion of insulin.

The oral administration of sodium fluoride (NaF) (40 mumol/100 body weight [bw]) to fasting rats produced an immediate fall in insulin levels and the consequent increase in glycemia. These phenomena were observed with plasma fluoride concentrations 5-15 microM. Glycemia and insulin returned to normal levels within 4-5 hours, together with the washing out of fluoride from plasma and soft tissues. The insulin secretion of isolated Langerhans islets, perifused with solutions containing 5, 10, or 20 microM fluoride, was found to be significantly inhibited as a function of fluoride levels, both with basal and stimulatory concentrations of glucose. One hour after the intake of 60 mg of NaF, fasting human volunteers showed increased fluoride (5-15 microM) together with a significant fall of plasma insulin levels.