Pathways to epilepsy surgery in children with tuberous sclerosis complex-associated epilepsy.

BACKGROUND: Previous studies showed the efficacy of epilepsy surgery in carefully selected children with epilepsy associated with tuberous sclerosis complex. However, how this selection is conducted, and the characteristics of the patients brought to surgery are still poorly described. By conducting a multicentric retrospective cohort study covering the practice of the last twenty years, we describe the paths leading to epilepsy surgery in children with epilepsy associated with tuberous sclerosis complex. METHODS: We identified 84 children diagnosed with tuberous sclerosis complex and epilepsy by matching two exhaustive registries of genetic diseases and subsequent medical records reviews within two French neuropediatric and epilepsy centers. Demographic, clinical, longitudinal, and diagnostic and surgical procedures data were collected. RESULTS: Forty-six percent of the children were initially drug-resistant and 19% underwent resective surgery, most often before the age of four. Stereotactic electroencephalography was performed prior to surgery in 44% of cases. Fifty-seven and 43% of patients remained seizure-free one and ten years after surgery, respectively. In addition, 52% of initially drug-resistant patients who did not undergo surgery were seizure-free at the last follow-up. The number of anti-seizure medications required decreased in 50% of cases after surgery. Infantile spasms, intellectual disability, autism spectrum disorder or severe behavioral disorders were not contraindications to surgery but were associated with a higher rate of complications and a lower rate of seizure freedom after surgery. CONCLUSION: Despite the assumption of complex multifocal epilepsy and practical difficulties in young children with tuberous sclerosis complex, successful surgery results are comparable with other populations of patients with drug-resistant epilepsy, and a spontaneous evolution to drug-sensitive epilepsy may occur in non-operated patients.

Rasmussen's encephalitis: Early diagnostic criteria in children.

Rasmussen's encephalitis (RE) is a rare chronic inflammatory brain disorder resulting in progressive neurodegeneration in one cerebral hemisphere. The inflammatory process is accompanied by progressive loss of function of the affected hemisphere, associated with drug-resistant partial epilepsy. The diagnosis is based on a range of clinical, electroencephalographic, radiological and biochemical arguments, without any specific formal marker, which makes the diagnosis of the disease complex, especially in its initial phase. Seizures are refractory to anti-seizures medication (ASM) and to classical immunomodulatory treatments. These treatments are also ineffective to stop the degenerative process. Only surgical treatment with hemispherotomy (surgical disconnection of a cerebral hemisphere) allows definitive cessation of seizures but this leads to definitive motor and cognitive deficits. The etiology of RE is not known, but there is strong evidence for an immunopathogenic mechanism involving T-cell mediated immunity. The emergence of biotherapies targeting against various cytokines offers potential therapeutic perspectives. This disease is currently a real challenge in terms of: (i) early diagnosis, before the constitution of marked hemispheric atrophy and the appearance of neurological and cognitive consequences; (ii) recognition of incomplete form; (iii) therapeutic management due to advances in the field of targeted treatment of inflammation; (iv) surgery and recovery possibilities.

Severe neonatal seizures: From molecular diagnosis to precision therapy?

Early onset epileptic encephalopathies (EOEE) are heterogeneous group of severe epilepsies that still need to be better defined and characterized. On a genetic point of view, several dozen of genes have been associated with EOEE, and to date, it is difficult to find a common mechanism to explain EOEE. In this short review, we show that two mains genes are involved in EOEE: STXBP1 and KCNQ2. Focusing on KCNQ2 related EOEE, we show that a relatively similar phenotype can be related to various consequences of mutations on a single gene. This will probably challenge the treatment of EOEE patients.

Reduction of health risk factors through an adapted physical activity program in patients with breast cancer.

PURPOSE: After a breast cancer diagnosis, patients are at high risk of reducing their physical activity and gaining weight. Lack of physical activity and weight gain are known negative but modifiable prognostic factors. An observational study of a 3-month adapted physical activity (APA) program was performed to assess its effectiveness in improving physical activity level and reducing risk factors related to health during or after breast cancer treatments. METHOD: Height, weight, and waist circumference (WC) were measured at the beginning and end of the 26-session program. Body mass index (BMI) and WC to height ratio (WHtR) were calculated. Physical activity profile, aerobic capacity, and usual average daily energy expenditure were estimated. Median values were compared using nonparametric tests. RESULTS: Sixty-one (61) voluntary breast cancer patients attended 80% of the sessions. At baseline, median (minimum-maximum) BMI was 23.3 (16.1-36.8) kg.m(-2) and WC and WHtR showed metabolic risks. After 3 months, anthropometric data remained stable. Moderate physical activity significantly improved (+13 min/day) and sedentary tended to decrease (-18 min/day). CONCLUSION: A 3-month APA program allows patients to limit risk factors related to health such as physical inactivity and metabolic risks. This study reinforces the need to promote physical activity as early as possible in cancer patients' care.

[Childhood periodic syndromes]. / Les syndromes périodiques de l'enfance.

This review focuses on the so-called "periodic syndromes of childhood that are precursors to migraine", as included in the Second Edition of the International Classification of Headache Disorders. Three periodic syndromes of childhood are included in the Second Edition of the International Classification of Headache Disorders: abdominal migraine, cyclic vomiting syndrome and benign paroxysmal vertigo, and a fourth, benign paroxysmal torticollis is presented in the Appendix. The key clinical features of this group of disorders are the episodic pattern and intervals of complete health. Episodes of benign paroxysmal torticollis begin between 2 and 8 months of age. Attacks are characterized by an abnormal inclination and/or rotation of the head to one side, due to cervical dystonia. They usually resolve by 5 years. Benign paroxysmal vertigo presents as sudden attacks of vertigo, accompanied by inability to stand without support, and lasting seconds to minutes. Age at onset is between 2 and 4 years, and the symptoms disappear by the age of 5. Cyclic vomiting syndrome is characterized in young infants and children by repeated stereotyped episodes of pernicious vomiting, at times to the point of dehydration, and impacting quality of life. Mean age of onset is 5 years. Abdominal migraine remains a controversial issue and presents in childhood with repeated stereotyped episodes of unexplained abdominal pain, nausea and vomiting occurring in the absence of headache. Mean age of onset is 7 years. Both cyclic vomiting syndrome and abdominal migraine are noted for the absence of pathognomonic clinical features but also for the large number of other conditions to be considered in their differential diagnoses. Diagnostic criteria, such as those of the Second Edition of the International Classification of Headache Disorders and the North American Society for Pediatric Gastroenterology, Hepatology and Nutrition, have made diagnostic approach and management easier. Their diagnosis is entertained after exhaustive evaluations have proved unrevealing. The recommended diagnostic approach uses a strategy of targeted testing, which may include gastrointestinal and metabolic evaluations. Therapeutic recommendations include reassurance, both of the child and parents, lifestyle changes, prophylactic therapy (e.g., cyproheptadine in children 5 years or younger and amitriptyline for those older than 5 years), and acute therapy (e.g., triptans, as abortive therapy, and 10% glucose and ondansetron for those requiring intravenous hydration).

Bacillus licheniformis septicemia in a very-low-birth-weight neonate: a case report.

Nosocomial infections in neonatal intensive care units are a preoccupying issue. Bacillus sp. can be pathogenic in immuno-compromised hosts, including premature infants. Central catheters and mechanical ventilation are potential sources of infection. We report for the first time a case of Bacillus licheniformis bacteremia in a premature infant. Recovery necessitated treatment with vancomycin and cefotaxime in combination with removal of the central catheter.

Large enhancement of the sub-barrier fusion probability for a halo nucleus

The fusion-fission cross sections of the 4He+238U and 6He+238U systems have been measured, at Louvain-la-Neuve, for energies around and below the Coulomb barrier, using an array of Si detectors surrounding a UF4 target. The data taken with 4He are in good agreement with previous data and with the coupled channel fusion calculation performed with ECIS. The 6He data show a regular trend with a large enhancement below the barrier which is attributed to the halo structure of the 6He nucleus.

Identification and characteristics of a 37,000 M(r) insulin-like growth factor binding protein in canine serum.

Insulin-like growth factors (IGFs) are potent stimulators of cellular growth and their half-life and biological activity are regulated by specific IGF binding proteins (IGFBPs). Western ligand blots of non-reduced human, bovine, ovine and porcine sera reveal an IGFBP-2 band at approximately 34,000 M(r). However, canine sera appear to contain a unique 37,000 M(r) IGFBP and lack the 34,000 M(r) IGFBP-2 band. In order to identify and characterize the 37,000 M(r) IGFBP, adult canine serum was subjected to non-reducing SDS polyacrylamide gel electrophoresis (PAGE), transferred to nitrocellulose paper, followed by [125I]-IGF-1 ligand blotting or immunoblotting with commercially available IGFBP antibodies. The 37,000 M(r) canine IGFBP reacted with an anti-IGFBP-2 antibody indicating that it is a canine analogue of IGFBP-2. However, the large difference in apparent molecular size indicates that this is a unique molecular form of IGFBP-2. N- or O-glycanase treatment of canine sera did not alter the molecular size of canine IGFBP-2 indicating that it is not a glycosylated variant of the IGFBP. Subjecting canine sera to reducing SDS-PAGE followed by anti-IGFBP-2 western immunoblotting revealed that the actual molecular weight of the canine IGFBP-2 is similar to that of reduced IGFBP-2 from other species indicating similar peptide lengths. Thus, the increased non-reduced size of the canine 37,000 M(r) IGFBP-2 is possibly due to a unique secondary structure.

Effect of colostrum intake on plasma glucose, non-esterified fatty acid and glucoregulatory hormone patterns in the neonatal pig.

The effect of colostrum on endocrine and metabolic factors affecting glucose homeostasis was evaluated in 60 neonatal pigs that were fasted, fed (nursed ad libitum) or limit-fed colostrum (25% ad libitum, 4-hr interval feeding). Plasma acquired at birth (t0), and after 10, 20 and 30 hr (t10, t20 and t30, respectively) was analyzed for glucose, non-esterified fatty acids (NEFA), and the glucoregulatory hormones--insulin, glucagon, cortisol, growth hormone and catecholamines. The concentration of glucose and NEFA was similar among treatment groups at birth and increased in proportion to the quantity of colostrum consumed. Pigs fed ad libitum achieved and maintained a higher (greater than or equal to 40%; P less than .01) glucose concentration when compared to fasted neonates. Limit-fed counterparts also achieved and maintained higher levels, with glucose concentration being approximately 20% higher throughout (P less than .05). Fed pigs maintained NEFA concentrations which were approximately 2.5-fold to 4-fold greater than that of fasted pigs (P less than .05). Likewise, limit-fed pigs tended (P = .19) to have elevated NEFA concentrations and a lower (P less than .05) insulin:glucagon molar ratio. An inverse relationship was observed between colostrum intake and plasma concentrations of cortisol and growth hormone. Concentrations of epinephrine and norepinephrine tended (P greater than .10) to be elevated in fed pigs, relative to those of fasted counterparts. Provision of even limited quantities of colostrum is therefore beneficial to the glucoregulatory response in newborn pigs.

Effect of pig weaning weight and associated nursery feeding programs on subsequent performance to 105 kilograms body weight.

An experiment using a total of 210 crossbred pigs from two farrowing groups evaluated the effects of three weaning weights and their associative starter feeding program on subsequent postweaning performance to 105 kg BW. One group of pigs nursed their dams in outside heated hutches (Trial 1), and a second group was raised in a centrally heated farrowing house (Trial II). The three pig weaning weight groups ranged in weight from 1) 4.1 to 5.0 kg, 2) 5.5 to 6.8 kg, and 3) 7.3 to 8.6 kg. Pigs in Group 1 were fed a high nutrient dense diet (HNDD) for a 2-wk period followed by a corn-soybean meal-dried whey (C-SBM-DW) and then a corn-soybean meal (C-SBM) diet, each for a 2-wk period. Group 2 was fed the same diet sequence except that HNDD was provided for 1 wk, whereas Group 3 was provided only the C-SBM-DW and the C-SBM diets each for 2 wk, consecutively. At the end of the nursery period, pigs were fed C-SBM diet formulations to 105 kg BW. Pigs of Trial I averaged 5.2 d older at weaning than those raised in the central farrowing house, but only a 1.5-d difference existed between light- and heavy-weight groups in both trials. Gains and feed intakes for the three weaning groups were higher as weaning weight increased during both the nursery and the growing-finishing period. There did not seem to be a compensatory growth response for lighter-weight weanling pigs even though they had been fed starter diets containing milk products. Consequently, fewer days (approximately 15) were required for the heavier-weight weanling pig group to reach a final weight of 105 kg than for the light-weight group. The medium-weight group required an intermediate number of days to reach 105 kg.

Efficacy of magnesium-L-ascorbyl-2-phosphate as a vitamin C source for weanling and growing-finishing swine.

Two experiments were conducted, one with weanling pigs (n = 288) and the second with grower-finisher swine (n = 216), to evaluate the efficacy of dietary vitamin C on various performance and serum measurements. Magnesium-L-ascorbyl-2-phosphate (46% L-ascorbic acid) served as the vitamin C source and was incorporated at dietary levels of 0, 50, or 500 ppm in both experiments. Pigs were allotted by sex, weight, and litter to randomized complete block designs. The nursery trial was conducted at four time periods and contained 12 replicates, whereas the grower-finisher experiment was over four time periods and contained nine replicates. Blood samples were collected initially from nine randomly selected pigs in both experiments, and from each pig within each pen at 2 and 5 wk postweaning, and at the 4- and 8-wk period in the grower-finisher trial. A killed Salmonella typhinurium bacterin was injected i.m. into starter pigs at 2 wk postweaning and at wk 4 and 6 in grower-finisher pigs. Hemagglutination titers were evaluated at 5 wk with the nursery pigs and at the 8-wk period with the grower-finisher swine. At the end of the grower-finisher trial, liver and kidney tissue were analyzed for ascorbate. Starter pigs grew faster (P < .05) and had improved gain:feed ratios (P < .05) when vitamin C was provided during the first 2 wk postweaning, but not during the latter 3-wk period. There was no improvement in pig gain or feed efficiency to vitamin C supplementation during any phase of the grower-finisher period.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of vitamin E, phosphorus and sorbitol on growth performance and serum and tissue cholesterol concentrations in the pig.

A total of 180 crossbred, weanling pigs were assigned to five dietary treatment groups: 1) a basal corn-soybean meal diet formulated to current NRC recommendations, 2) basal + monosodium phosphate (2 x NRC P recommendations; P), 3) basal + alpha-tocopheryl acetate (220 IU/kg; E), 4) basal + sorbitol (1% of the diet; S) and 5) basal + PES. Dietary treatments were continued until market weight (104 kg). Blood samples were obtained at 3-wk intervals for analysis of serum alpha-tocopherol, P and total cholesterol. Liver and muscle (semimembranosus) samples were obtained at the end of the starter, grower and finisher phases for determination of total cholesterol concentration. The Ca:P imbalance produced by the high-phosphorus diets (P and PES) increased feed intake during the finisher phase. Dietary treatment did not consistently affect total serum cholesterol at any phase of growth. A transient 21.5% (P less than .05) depression of liver cholesterol concentration was observed in the PES-fed pigs at the end of the starter phase but was not apparent at market weight. A similar trend (nonsignificant) was noted for muscle cholesterol concentration. The present study suggests that the PES diet can decrease tissue cholesterol concentration during the nursery phase, but it remains uncertain whether this transient response is a function of age and(or) diet transition at weaning. Further research is necessary to determine whether this response can be translated to the finishing phase, and thereby reduce carcass cholesterol.

Efficacy of dietary D-alpha-tocopherol and DL-alpha-tocopheryl acetate for weanling pigs.

A 2 x 3 factorial experiment in a randomized complete block design was conducted using a total of 180 weanling pigs in five replicates. The study evaluated the efficacy of two dietary vitamin E sources (D-alpha-tocopherol, DL-alpha-tocopheryl acetate) added at three dietary levels (16, 48, 96 IU/kg) during a 35-d postweaning trial. Pigs within each treatment were fed two similarly fortified vitamin E diets in sequence; the first contained 40% milk products and was fed to 14 d, and the second contained 20% milk product and 5% fat and was provided from 15 to 35 d postweaning. Five pigs per pen per replicate were bled weekly for serum analysis of alpha-tocopherol, Se, cholesterol, triglyceride, and glutathione peroxidase (GSH-Px) activity. At the end of the trial, one pig per pen was randomly selected and killed with liver, loin, lung, and heart excised and frozen for tocopherol analysis. Postweaning gains, feed intakes, and efficiencies were similar between the two vitamin E sources and at the various dietary levels. Serum tocopherol concentrations were consistently higher when D-alpha-tocopherol was provided. Vitamin E sources and levels had no effect nor did they influence weekly serum Se, cholesterol, or triglyceride concentrations or GSH-Px activity. A serum and tissue interaction (P less than .05) response occurred between dietary vitamin E source x level with alpha-tocopherol concentrations increasing linearly (P less than .01) as dietary vitamin E level increased, but at a higher rate when D-alpha-tocopherol than when DL-alpha-tocopheryl acetate as fed.(ABSTRACT TRUNCATED AT 250 WORDS)

Growth performance of weanling pigs fed corn-soybean meal diets with or without dried whey at various L-lysine.HCl levels.

A total of 490 crossbred weanling pigs were used to evaluate the responses to and the subsequent interaction between dietary dried whey and crystalline L-lysine.HCl on postweanling growth and feed efficiency at two periods postweaning. The experiment was conducted as a 2 x 5 factorial arrangement of treatments in a randomized complete block design to evaluate two levels of edible-grade dried whey (0 or 25%) and five dietary lysine levels ranging from 1.10 to 1.50% in .10% increments using a corn-soybean meal mixture as the basal feedstuff. Pigs were allotted by weight, litter, and sex to seven replicates at weaning (23 +/- 2 d) and fed their treatment diets for a 35-d period. Daily gain and feed intake were greater (P less than .01) for both the 0- to 21- and the 22- to 35-d periods when dried whey was fed; the relative magnitude of the response to dried whey was greatest during the initial 21-d period. Growth responses during the 0- to 21-d period were, however, independent of dietary lysine level, suggesting that dietary lysine at a level of 1.10% is not the limiting nutrient in a corn-soybean meal diet or a corn-soybean meal diet with dried whey. From 22 to 35 d postweaning a linear growth response to lysine level occurred when the dried whey diet was fed, but no response was detected when lysine was added to the corn-soybean diet, resulting in a diet x lysine level interaction (P less than .10).(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of colostrum intake on hepatic gluconeogenesis and fatty acid oxidation in the neonatal pig.

A total of 24 newborn pigs were used to determine 1) the relationship between the quantity of colostrum consumed and the capacity for gluconeogenesis and fatty acid oxidation and 2) whether fatty acid oxidation limits gluconeogenesis in isolated hepatocytes. Neonatal pigs were obtained prior to nursing and allotted to one of three treatment groups; fed (ad libitum), limit-fed (25% of fed group), or fasted. Hepatocytes were isolated when pigs were 24 h old. Colostrum intake altered the metabolic status of neonates such that the capacity for glucose synthesis and oxidation of octanoate increased with intake. Glucose synthesis with lactate as the substrate was greater (P less than .01) for fed pigs (10.79 mumol glucose.h-1.mg DNA-1) than for either limit-fed (6.56) or fasted counterparts (4.78), which were similar (P greater than .10). Colostrum intake failed to stimulate synthesis from alanine. The oxidation rate for octanoate was similar for fed and limit-fed pigs (.62 and .61 nmol CO2.h-1.mg DNA-1, respectively) but greater (P less than .05) than that observed for fasted counterparts (.36). Oxidation of octanoate (2 mM) was approximately 30-fold greater than for oleate (1 mM); oxidation of the latter was not affected by either colostrum intake or the addition of carnitine (1 mM). The increase in octanoate oxidation, however, did not elicit an increase in glucose synthesis by fasting pigs with either lactate or alanine as precursors. Thus, we conclude that gluconeogenesis is a function of colostrum intake and that reducing equivalents and(or) ATP may not be primary factors limiting glucose synthesis in pigs fasted from birth.

Dietary clofibric acid increases intestinal fatty acid binding protein activity and apparent lipid digestibility in weanling swine.

The effects of supplementation of dietary clofibric acid (.5% wt/wt) on fatty acid binding protein (FABP) activity, apparent lipid digestibility, and serum cholesterol concentrations were evaluated in weanling pigs. Twenty-four barrows were allotted by weight and litter to a randomized complete block design with two treatments (basal vs clofibric acid) in six replicates. Nutrient digestibility measurements were made for a 2-wk period, after which the pigs were killed and tissues were collected. No differences in BW, liver, proximal small intestine, distal small intestine, and proximal and distal intestinal mucosa weights were observed. Apparent lipid digestibility was greater (P < .05) for the overall 2-wk period in clofibric acid-supplemented pigs (81.5 vs 76.6%). This paralleled the increased FABP activity in the distal small intestine (P < .001) of clofibric acid-supplemented pigs. Proximal intestine and liver FABP activities were unaffected by dietary treatment. Serum cholesterol concentrations were markedly lowered by clofibric acid supplementation. During wk 1, pigs fed the basal diet had twofold greater (P < .01) serum cholesterol concentrations, whereas during wk 2, basal-fed pigs had fourfold greater (P < .01) serum cholesterol concentrations (81.5 vs 18.3 mg/dL). These results suggest that elevated intestinal FABP activities may augment fatty acid absorption from the gastrointestinal tract.

Effect of colostrum or medium-chain triglyceride supplementation on the pattern of plasma glucose, non-esterified fatty acids and survival of neonatal pigs.

A total of 48 neonatal pigs were used to determine whether intubation with colostrum or medium-chain triglyceride (MCT) would enhance glucose homeostasis and survival. Pigs were removed from the sow prior to nursing and alloted to three treatment groups. Fasted pigs received only water for 30 h, whereas those allotted to supplemented groups received either 30 ml of colostrum or 15 ml of MCT at 6 and 16 h after birth (t6 and t16). Supplementation with MCT (t6) resulted in a 2.4-fold elevation in plasma non-esterified fatty acid (NEFA) concentration compared with fasted pigs (260 vs 109 microEq/liter at t8; P less than .05). This difference increased following the second MCT dose (436 vs 117 microEq/liter at t18; P less than .05). Colostrum supplementation also elevated plasma NEFA (201 and 259 microEq/liter at t8 and t18, respectively); however, less triglyceride fatty acid was presented via colostrum compared with MCT. Supplementation with MCT resulted in a greater increase in plasma glucose concentration, relative to fasting levels (75 vs 56 mg/ml at t8; 76 vs 62 mg/ml at t18), than was obtained with colostrum (68 and 65 mg/ml at t8 and t18, respectively). Residual effects of supplementation to t30 were evident for both MCT and colostrum pigs in NEFA levels, but only the MCT group had a greater (P less than .05) concentration of plasma glucose at t30 compared with the fasted group (63 vs 49; P less than .05). This regimen of MCT supplementation was employed in an experiment with nursing pigs to determine whether MCT would improve glucose status and survival of less-competitive pigs.(ABSTRACT TRUNCATED AT 250 WORDS)

Dietary nitrogen and lipid utilization by growing pigs fed structured triacylglycerides synthesized from medium-chain triacylglycerides and menhaden oil.

A total of 24 crossbred barrows were used in a 19-d metabolism trial to determine the effect of dietary structured triacylglycerides synthesized by the random reesterification of medium-chain triacylglycerides (MCT) and menhaden oil on growth performance, nitrogen retention, and apparent digestibilities of nitrogen, lipid, and fatty acids. Pigs were randomly assigned to four dietary treatments differing only in lipid source. Dietary treatments (percentage by weight of total lipid) contained either 1) corn oil: soybean oil:MCT oil (40:10:50), 2) MCT oil:menhaden oil (60:40 as structured triacylglyceride), 3) MCT oil: menhaden oil (60:40 nonstructured, physical mixture), or 4) structured triacylglyceride (as in 2): safflower oil:canola oil (80:10:10). Total fecal and urine collections were conducted during two 5-d periods. Apparent nitrogen digestibility and nitrogen retention were high (> 95%) and not affected (P > .05) by dietary treatment. Apparent fatty acid digestibilities were affected by dietary lipid source. Palmitic and arachidonic acid digestibilities were lower (P < .05) for Diet 1, heptadecanoic acid digestibility was elevated by 21% (P < .05) for Diet 2 relative to Diet 3, and stearic acid digestibility was highest (P < .05) for Diet 1, followed by Diets 2, 3, and 4 in decreasing order. This study demonstrates that the physical nature of the lipid (structured vs nonstructured) affects fatty acid digestibility patterns in the growing pig. Further evaluation of MCT/menhaden oil structured triacylglyceride is needed.

Beta-carotene uptake and changes in ovarian steroids and uterine proteins during the estrous cycle in the canine.

The uptake of beta-carotene by reproductive tissues and the effects of beta-carotene on reproductive function in the dog are unknown. We studied the uptake of beta-carotene by blood, corpus luteum, and uterine endometrium and the role of dietary beta-carotene in influencing ovarian steroid and uterine protein production during the estrous cycle in the dog. Mature female Beagle dogs (n = 56) were fed diets containing 0, 2, 20, or 50 mg of beta-carotene daily for approximately 6 wk before estrus detection. Blood was sampled at regular intervals from estrus through d 45 after ovulation (d 0 = ovulation), when laparotomy was performed. The ovaries were obtained for the isolation of corpus luteum. The uterus was flushed with phosphate-buffered saline and the endometrium obtained by scraping. Beta-carotene was not detectable in plasma, corpus luteum, or endometrium of unsupplemented dogs. However, beta-carotene and alpha-carotene in plasma, corpus luteum, and uterine endometrium increased in a dose-dependent manner. Alpha-carotene made up a high percentage of total carotenoids even though the alpha-carotene content in the dietary source was very low. Dogs fed 50 mg of beta-carotene had significantly higher concentrations of plasma progesterone between d 12 and 26 compared with unsupplemented dogs. Dietary beta-carotene did not influence plasma estradiol-17beta and total uterine proteins. Therefore, beta-carotene is absorbed into plasma, corpus luteum, and uterine endometrium of dogs. Furthermore, dietary beta-carotene increased plasma progesterone concentrations during the estrous cycle. It is possible that dietary beta-carotene may improve reproductive function in the canine.

Quantitative analysis of chondroitin sulfate isomers in intervertebral disk chondrocyte culture using capillary electrophoresis.

Chondroitin sulfate (CS) isomers, 6-sulfate (CS6) and 4-sulfate (CS4), change their ratio to each other in cartilaginous tissues with aging. In this study, a quantitative measurement method of CS6 and CS4 was developed, using capillary electrophoresis (CE). Various buffer solutions, pH, and digestion times were studied, and the use of 0.1 M Tris-HCl at pH of 8.0 allowed the isolation of CS6 and CS4 from CS most efficiently when combined with chondrotinase ABC at a concentration of 1 mU/microg of the substrate during a 3 hr digestion period. Amounts of newly synthesized CS6 and CS4 in the intervertebral disk chondrocyte three-dimensional culture were quantified by this method after the proteoglycans were extracted by equilibrium density centrifugation.