Analysis of acylcarnitine profiles in umbilical cord blood and during the early neonatal period by electrospray ionization tandem mass spectrometry

Acylcarnitine profiling by electrospray ionization tandem mass spectrometry (ESI-MS/MS) is a potent tool for the diagnosis and screening of fatty acid oxidation and organic acid disorders. Few studies have analyzed free carnitine and acylcarnitines in dried blood spots (DBS) of umbilical cord blood (CB) and the postnatal changes in the concentrations of these analytes. We have investigated these metabolites in healthy exclusively breastfed neonates and examined possible effects of birth weight and gestational age. DBS of CB were collected from 162 adequate for gestational age neonates. Paired DBS of heel-prick blood were collected 4-8 days after birth from 106 of these neonates, the majority exclusively breastfed. Methanol extracts of DBS with deuterium-labeled internal standards were derivatized before analysis by ESI-MS/MS. Most of the analytes were measured using a full-scan method. The levels of the major long-chain acylcarnitines, palmitoylcarnitine, stearoylcarnitine, and oleoylcarnitine, increased by 27, 12, and 109%, respectively, in the first week of life. Free carnitine and acetylcarnitine had a modest increase: 8 and 11%, respectively. Propionylcarnitine presented a different behavior, decreasing 9% during the period. The correlations between birth weight or gestational age and the concentrations of the analytes in DBS were weak (r £ 0.20) or nonsignificant. Adaptation to breast milk as the sole source of nutrients can explain the increase of these metabolites along the early neonatal period. Acylcarnitine profiling in CB should have a role in the early detection of metabolic disorders in high-risk neonates.

Analysis of acylcarnitine profiles in umbilical cord blood and during the early neonatal period by electrospray ionization tandem mass spectrometry.

Acylcarnitine profiling by electrospray ionization tandem mass spectrometry (ESI-MS/MS) is a potent tool for the diagnosis and screening of fatty acid oxidation and organic acid disorders. Few studies have analyzed free carnitine and acylcarnitines in dried blood spots (DBS) of umbilical cord blood (CB) and the postnatal changes in the concentrations of these analytes. We have investigated these metabolites in healthy exclusively breastfed neonates and examined possible effects of birth weight and gestational age. DBS of CB were collected from 162 adequate for gestational age neonates. Paired DBS of heel-prick blood were collected 4-8 days after birth from 106 of these neonates, the majority exclusively breastfed. Methanol extracts of DBS with deuterium-labeled internal standards were derivatized before analysis by ESI-MS/MS. Most of the analytes were measured using a full-scan method. The levels of the major long-chain acylcarnitines, palmitoylcarnitine, stearoylcarnitine, and oleoylcarnitine, increased by 27, 12, and 109%, respectively, in the first week of life. Free carnitine and acetylcarnitine had a modest increase: 8 and 11%, respectively. Propionylcarnitine presented a different behavior, decreasing 9% during the period. The correlations between birth weight or gestational age and the concentrations of the analytes in DBS were weak (r ≤ 0.20) or nonsignificant. Adaptation to breast milk as the sole source of nutrients can explain the increase of these metabolites along the early neonatal period. Acylcarnitine profiling in CB should have a role in the early detection of metabolic disorders in high-risk neonates.

Effect of TNF-α production inhibitors on the production of pro-inflammatory cytokines by peripheral blood mononuclear cells from HTLV-1-infected individuals

Human T lymphotropic virus type 1 (HTLV-1) is the causal agent of myelopathy/tropical spastic paraparesis (HAM/TSP), a disease mediated by the immune response. HTLV-1 induces a spontaneous proliferation and production of pro-inflammatory cytokines by T cells, and increasing interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) levels are potentially involved in tissue damage in diseases related to HTLV-1. This exaggerated immune response is also due to an inability of the natural regulatory mechanisms to down-modulate the immune response in this group of patients. TNF-α inhibitors reduce inflammation and have been shown to improve chronic inflammatory diseases in clinical trials. The aim of this study was to evaluate the ability of pentoxifylline, forskolin, rolipram, and thalidomide to decrease in vitro production of TNF-α and IFN-γ in cells of HTLV-1-infected subjects. Participants of the study included 19 patients with HAM/TSP (mean age, 53 ± 11; male:female ratio, 1:1) and 18 HTLV-1 carriers (mean age, 47 ± 11; male:female ratio, 1:2.6). Cytokines were determined by ELISA in supernatants of mononuclear cell cultures. Pentoxifylline inhibited TNF-α and IFN-γ synthesis with the minimum dose used (50 µM). The results with forskolin were similar to those observed with pentoxifylline. The doses of rolipram used were 0.01-1 µM and the best inhibition of TNF-α production was achieved with 1 µM and for IFN-γ production it was 0.01 µM. The minimum dose of thalidomide used (1 µM) inhibited TNF-α production but thalidomide did not inhibit IFN-γ production even when the maximum dose (50 µM) was used. All drugs had an in vitro inhibitory effect on TNF-α production and, with the exception of thalidomide, all of them also decreased IFN-γ production.

Effect of TNF-α production inhibitors on the production of pro-inflammatory cytokines by peripheral blood mononuclear cells from HTLV-1-infected individuals.

Human T lymphotropic virus type 1 (HTLV-1) is the causal agent of myelopathy/tropical spastic paraparesis (HAM/TSP), a disease mediated by the immune response. HTLV-1 induces a spontaneous proliferation and production of pro-inflammatory cytokines by T cells, and increasing interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) levels are potentially involved in tissue damage in diseases related to HTLV-1. This exaggerated immune response is also due to an inability of the natural regulatory mechanisms to down-modulate the immune response in this group of patients. TNF-α inhibitors reduce inflammation and have been shown to improve chronic inflammatory diseases in clinical trials. The aim of this study was to evaluate the ability of pentoxifylline, forskolin, rolipram, and thalidomide to decrease in vitro production of TNF-α and IFN-γ in cells of HTLV-1-infected subjects. Participants of the study included 19 patients with HAM/TSP (mean age, 53 ± 11; male:female ratio, 1:1) and 18 HTLV-1 carriers (mean age, 47 ± 11; male:female ratio, 1:2.6). Cytokines were determined by ELISA in supernatants of mononuclear cell cultures. Pentoxifylline inhibited TNF-α and IFN-γ synthesis with the minimum dose used (50 µM). The results with forskolin were similar to those observed with pentoxifylline. The doses of rolipram used were 0.01-1 µM and the best inhibition of TNF-α production was achieved with 1 µM and for IFN-γ production it was 0.01 µM. The minimum dose of thalidomide used (1 µM) inhibited TNF-α production but thalidomide did not inhibit IFN-γ production even when the maximum dose (50 µM) was used. All drugs had an in vitro inhibitory effect on TNF-α production and, with the exception of thalidomide, all of them also decreased IFN-γ production.

[Salivary and lacrymal gland disorders and HTLV-1 infection]. / Atteinte des glandes salivaires et lacrymales et infection par le HTLV-1.

INTRODUCTION: The frequency and severity of salivary and lacrymal gland human T-cell lymphotropic virus type 1 (HTLV-1) infection were assessed in HTLV-1 plus patients, presenting with neurological deficit (tropical spastic paraparesis/HTLV-1 associated myelopathy [TSP/HAM]) or not. The mechanism of this deficit was investigated. MATERIAL AND METHODS: A case-control study was made from April 2002 to December 2005, in an area strongly endemic for HTLV-1. The patients were classified in three groups: group 1 with 16 patients presenting with TSP/HAM; group 2 with 67 HTLV-1 carriers and group 3 with 29 healthy volunteers. The dry syndrome was investigated by history taking and by oral and ophthalmological clinical examination. Immunological and biological screening for rhumatoid factors, antinuclear antibodies, and antibodies against soluble nuclear antigens (SSA, SSB). Peripheral blood was separated by density gradient and mononuclear cells were recovered to dose interferon-gamma and tumor necrosis factor-alpha. Patients in the three groups were assessed for salivary flow by stimulated weighing using Saxon's test. A Chi-2 test, a variance analysis (Anova), and the Spearman rank correlation test were used for the statistical analysis. RESULTS: The dry syndrome was mild and more common in group 1 patients (75%). In group 2, 22% of the patients presented with functional signs of buccal mucosa dryness comparable to those observed in group 1. No correlation was found between salivary flow and screened pro-inflammatory cytokines. DISCUSSION: Our results show that hyposialia is an important part of the disease induced by HTLV-1, even in virus carriers without neurological deficit. Its mechanism seems different than that of the Gougerot-Sjögren syndrome.

Linear growth and zinc supplementation in children with short stature.

Physical growth retardation is an early and prominent feature of zinc deficiency, but the effect of zinc supplementation in children is still not completely clear. This study investigated the impact of zinc supplementation on linear growth, growth velocity, IGF-I levels, and skeletal maturation of short children during and after mineral supplementation. The study was designed as a double-blind, randomized, controlled trial of zinc supplementation during a 6-month period, with a subsequent 6-month follow-up. Anthropometric data were collected at 0, 6, and 12 months. Measurements included plasma Zn, IGF-I, height, weight, triceps skinfold thickness, and body mass index. Eighteen healthy pre-pubertal short children (z-score -2.0) 7 to 10 years old with normal GH and IGF-I levels were randomized to two groups, one with zinc supplementation (5 mg/kg/d of ZnSO4) and the other with placebo. In the first 6 months, only height velocity increased significantly, 5.99+/-0.80 cm/yr vs 5.05+/-0.85 cm/yr (p=0.03). After 12 months, height velocity returned to the initial values, 3.92+/-0.59 cm/yr vs 4.19+/-1.08 cm/yr (p=0.29). This study indicates that zinc supplementation increased growth velocity, but these effects did not persist after supplementation was discontinued.

[Blood circulation, or the movement in the concept of motion]. / A circulação do sangue, ou o movimento no conceito de movimento.

The contemporary medical practice is enrolled in the rigid principles of a technological order. In order that the development and organization of the technological order would take place, and also material and objetively made concrete, it was essential that their products represented a materialization of a certain concept of human body and of health. This paper intends to be an introductory study of certain conditions of possibility for the construction of this technological order in the area of health. Although this thought includes, necessarily, a historical dimension, we do not intend here a factual reconstitution that considers dates and personalities. Its immediate goal will be to focus a moment of rupture, in which the introduction of the new concept of motion generated a new conception of human physiology. So, more important than one specific theory--of the blood circulation--or its effective results, is to understand the process of arising and/or changing of the concepts that allowed it to exist originally.

Tocolysis with indomethacin increases the incidence of necrotizing enterocolitis in the low-birth-weight neonate.

OBJECTIVE: The null hypothesis states that prolonged antenatal indomethacin exposure within 24 hours of delivery does not increase the incidence of necrotizing enterocolitis in the low-birth-weight neonate. STUDY DESIGN: The neonates of patients receiving indomethacin tocolysis admitted in preterm labor (N = 56) were compared with the neonates of preterm labor patients who received no indomethacin tocolysis (N = 703). These neonatal groups were then compared with regard to gestational age at delivery, birth weight, mode of delivery, antenatal magnesium and steroid exposure, incidence of respiratory distress syndrome, perinatal depression, sepsis, umbilical catheterization, and feeding rates and volumes. The overall incidence of necrotizing enterocolitis, mortality secondary to necrotizing enterocolitis, and the intervals from delivery and feeding to necrotizing enterocolitis diagnosis were also compared. The association between necrotizing enterocolitis and the duration of indomethacin exposure and the interval from exposure to delivery for both the indomethacin and control groups was determined. RESULTS: The incidence of necrotizing enterocolitis in neonates who were delivered within 24 hours of maternal indomethacin therapy was 20% compared with 9% in the control group (p = 0.005). The incidence of necrotizing enterocolitis in neonates with > 48 hours of antenatal indomethacin exposure was 26.4% compared with 4.1% in those with < 48 hours exposure (p = 0.042). The interval from first feeding to necrotizing enterocolitis development was significantly shorter in the indomethacin group versus the control group (2.1 +/- 3.0 vs 6.8 +/- 6.3 days) (p = 0.001), as was the mean interval from delivery to development of necrotizing enterocolitis (10.2 +/- 3.7 vs 15.2 +/- 3.8 days) (p = 0.019). CONCLUSIONS: Antenatal indomethacin exposure occurring within < or = 24 hours of delivery and of at least 48 hours' duration is associated with a significant increase in the incidence of necrotizing enterocolitis in the low-birth-weight neonate.

Blood platelets and angiotensin II: angiotensin II release after platelet aggregation.

Recent studies have demonstrated the presence of angiotensin II (Ang II) in human platelets. The aim of the present study was to determine whether in vitro platelet aggregation induces Ang II release. We studied 20 healthy volunteers, and found that some aggregant stimuli can cause the platelet to release this peptide. Release of Ang II was greater with thrombin (40.7%) than with ADP (29%), while N-ethylmaleimide was almost ineffective. The release of Ang II following in vitro aggregant stimuli suggests that this may be one of the mechanisms through which platelets can locally modulate vascular tone and perhaps promote atherogenesis.

Effect of nicardipine on angiotensin II platelet content.

We evaluated the effect of nicardipine treatment on platelet angiotensin II content in 18 essential hypertensive patients. Nicardipine induced a decrease in platelet angiotensin II content in all patients (-14%), but particularly in 10 (P less than 0.05; -20.5%). No other differences were found between these patients and the others. Further investigations are required to determine whether platelet angiotensin II content reduction contributes to the prophylactic and therapeutic effects of calcium antagonists on atherosclerosis.

Effect of angiotensin converting enzyme inhibition on platelet angiotensin II content.

The effect of captopril treatment on platelet angiotensin II levels was evaluated in 12 patients with essential hypertension. Captopril significantly lowered (p less than 0.01) mean arterial blood pressure in seven patients (Group 1) and was ineffective in five (Group 2). In Group 1, a marked decrease in plasma angiotensin II levels in both the supine (from 5.5 +/- 0.5 to 2.8 +/- 0.9 pg/ml) and the upright positions (from 17.5 +/- 4.7 to 3.9 +/- 1.6 pg/ml; p less than 0.0025) and a significant increase in platelet angiotensin II levels (from 10.5 +/- 5.3 to 22.4 +/- 17 pg/ml; p less than 0.05) after captopril treatment was observed. In Group 2, no variation was found in plasma angiotensin II levels, whereas platelet angiotensin II levels increased slightly (from 10.8 +/- 3.1 pg/ml to 16.9 +/- 5.2 pg/ml; NS). These findings suggest that the decrease in plasma angiotensin II levels can lead to an increase in platelet angiotensin II receptors or can lead to angiotensin II production in platelets through activation of a feedback mechanism. The second hypothesis suggests that platelets have alternative enzymatic pathways leading to angiotensin II production not inhibited by captopril.