Adipose tissue: a new target for electroporation-enhanced DNA vaccines.

DNA vaccines delivered using electroporation (EP) have had clinical success, but these EP methods generally utilize invasive needle electrodes. Here, we demonstrate the delivery and immunogenicity of a DNA vaccine into subcutaneous adipose tissue cells using noninvasive EP. Using finite element analysis, we predicted that plate electrodes, when oriented properly, could effectively concentrate the electric field within adipose tissue. In practice, these electrodes generated widespread gene expression persisting for at least 60 days in vivo within interscapular subcutaneous fat pads of guinea pigs. We then applied this adipose-EP protocol to deliver a DNA vaccine coding for an influenza antigen into guinea pigs. The resulting host immune responses elicited were of a similar magnitude to those achieved by skin delivery with EP. The onset of the humoral immune response was more rapid when the DNA dose was spread over multiple injection sites, and increasing the voltage of the EP device increased the magnitude of the immune response. This study supports further development of EP protocols delivering gene-based therapies to subcutaneous fat.

Epicardial fat thickness in infants of diabetic mothers.

BACKGROUND: Epicardial fat thickness (EFT) in adults and children has been related to the metabolic syndrome and other cardiovascular risk factors; however, scarce studies have evaluated it in infants of diabetic mothers (IDM) in whom, alterations in the thickness of the interventricular septum have been reported. This study compares the EFT in IDM versus infants of non-diabetic mothers (INDM) and its association with others echocardiographic parameters. METHODS: We performed a cross sectional study in 93 infants (64 IDM and 29 INDM). To evaluate EFT dimensions, an echocardiogram was performed within the first 24h of extrauterine life in both groups. In diabetic mothers, HbA1c was also determined. RESULTS: There was no significant difference in birth weight between the groups although gestational age was lower in IDM. The EFT (3.6 vs. 2.5 mm, p < 0.0001), the interventricular septum thickness (IVST) (6.2 vs. 5.2 mm, p < 0.0001) and the IVST / left ventricle posterior wall (1.3 vs. 1.1, p = 0.001) were higher in the IDM; while the left ventricular expulsion fraction [LVFE] (71.1 vs. 77.8; p < 0.0001) was lower than in the INDM, respectively. We found a positive correlation between EFT with IVST (r = 0.577; p = 0.0001), LVPW (r = 0.262; p = 0.011), IVST/LVPW index (r = 0.353; p = 0.001), and mitral integral early velocity (r = 0.313; p = 0.002), while a negative correlation with LVFE was observed (r = -0.376; p = 0.0001). CONCLUSIONS: The EFT is higher in IDM than in INDM. It was positively related with echocardiographic parameters of left ventricular thickness and negatively with left ventricular ejection function.

Age-growth relationships, temperature sensitivity and palaeoclimate-archive potential of the threatened Altiplano cactus Echinopsis atacamensis.

The tall (>4 m), charismatic and threatened columnar cacti, pasacana [Echinopsis atacamensis (Vaupel) Friedrich & G.D. Rowley)], grows on the Bolivian Altiplano and provides environmental and economic value to these extremely cold, arid and high-elevation (~4000 m) ecosystems. Yet very little is known about their growth rates, ages, demography and climate sensitivity. Using radiocarbon in spine dating time series, we quantitatively estimate the growth rate (5.8 and 8.3 cm yr-1) and age of these cacti (up to 430 years). These data and our field measurements yield a survivorship curve that suggests precipitation on the Altiplano is important for this species' recruitment. Our results also reveal a relationship between nighttime temperatures on the Altiplano and the variation in oxygen isotope values in spines (δ18O). The annual δ18O minimums from 58 years of in-series spine tissue from pasacana on the Altiplano provides at least decadal proxy records of temperature (r = 0.58; P < 0.0001), and evidence suggests that there are longer records connecting modern Altiplano temperatures to sea-surface temperatures (SSTs) in the Atlantic Ocean. While the role of Atlantic SSTs on the South American Summer Monsoon (SASM) and precipitation on the Bolivian Altiplano is well described, the impact of SSTs on Altiplano temperatures is disputed. Understanding the modern impact of SSTs on temperature on the Altiplano is important to both understand the impact of future climate change on pasacana cactus and to understand past climate changes on the Altiplano. This is the best quantitative evidence to date of one of the oldest known cactus in the world, although there are likely many older cacti on the Altiplano, or elsewhere, that have not been sampled yet. Together with growth, isotope and age data, this information should lead to better management and conservation outcomes for this threatened species and the Altiplano ecosystem.

Evidence-based recommendations for the management of ankylosing spondylitis: systematic literature search of the 3E Initiative in Rheumatology involving a broad panel of experts and practising rheumatologists.

OBJECTIVE: Recommendations and/or guidelines represent a popular way of integrating evidence-based medicine into clinical practice. The 3E Initiatives is a multi-national effort to develop recommendations for the management of rheumatic diseases, which involves a large number of experts combined with practising rheumatologists addressing specific questions relevant to clinical practice. METHODS: Ten countries participated in three rounds of discussions and votes concerning the management of AS. A set of nine questions was formulated in the domains of diagnosis, monitoring and treatment, after a Delphi procedure. A literature search in MedLine was conducted. Predefined outcome parameters for the domains of diagnosis, monitoring and treatment were assessed. The evidence to support each proposition was evaluated and scored. After discussion and votes, the final recommendations were presented using brief statements by each national group, following which the final international recommendations were formulated. RESULTS: A total of 2699 papers were found and 467 were selected for analysis. Twelve key recommendations were developed: three in the domain of diagnosis addressing general diagnostic considerations, early AS diagnosis and general practitioners' referral recommendations; three concerning monitoring of AS disease activity, severity and prognosis; six concerning pharmacological treatment (except biologics): non-steroidal anti-inflammatory drugs/COX-II inhibitors, bisphosphonates and treatment of enthesitis. The compiled agreement among experts ranged from 72% to 93%. CONCLUSION: Recommendations for the management of AS were developed using an evidence-based approach followed by expert/physician consensus with high level of agreement. Involvement of a larger and more representative group of rheumatologists may improve their dissemination and implementation in daily clinical practice.

Evaluation of the pretransplantation phase of the hemopoietic stem cell program in a tertiary hospital.

INTRODUCTION: Therapeutic decisions and clinical events during the pretransplantation phase of stem cell transplantation (SCT) may influence survival, quality of life, and efficiency of health expenses. However, there is a lack of relevant published data. AIMS: The aims of this study were to identify reasons why the procedure was not performed and to know the waiting time for SCT candidates. PATIENTS AND METHODS: We collected pretransplantation data from 166 consecutive patients evaluated by the SCT Committee of a tertiary center between April 2005 and December 2006. RESULTS: One hundred fifty-two of 166 patients were referred for the first time. Additionally, 14 were reconsidered as candidates for a subsequent SCT due to relapse, graft failure, secondary malignancy, or a multiple-graft program. One hundred forty-one were accepted for transplantation, whereas 25 were not. At the time of analysis, 22 patients were still awaiting SCT, 8 were delayed because they required additional courses of treatment, and 32 were excluded because of death (34.4%), poor stem cell mobilization (21.9%), patient refusal (15.6%), relapse/progression (9.4%), comorbidity (6.3%), or absence of a donor (6.3%). The median time between inclusion in the program and transplantation was 3.6 months (range, 0.27-13.43), and 5.7 months (P < .05) for unrelated allogeneic transplantation. No significant differences were observed in the diagnosis or hospital of origin. CONCLUSIONS: SCT was not performed in 22% of transplant candidates, mainly due to death, insufficient stem cell mobilization, patient refusal, or disease progression/relapse. The median time between inclusion in the SCT program and transplantation was 3 months, but longer among the unrelated allogeneic transplantations.

Association analysis of the Gln223Arg polymorphism in the human leptin receptor gene, and traits related to obesity in Mexican adolescents.

Polymorphisms of leptin receptor (LEPR) may contribute to a common form of obesity and, as a consequence, obesity-related diseases. We evaluated the potential role of genetic variation at the LEPR gene in heart sympathetic activity and other traits related to obesity in Mexican adolescents. Adolescents aged between 12 and 17 years, with steady body weight for the last 3 months were included. We evaluated anthropometric measurements, blood pressure, seric glucose, insulin, leptin levels, heart sympathetic activity (by electrocardiograph monitoring at rest), and the Gln223Arg and Pro1019Pro LEPR polymorphisms in each subject. In total, 103 adolescents (55 obese and 48 nonobese) were included. The group of obese adolescents showed higher sympathetic activity, blood pressure, glucose, insulin, and leptin levels. The genotype frequencies for the two polymorphisms were found to be in Hardy-Weinberg equilibrium. There was no difference in the genotype frequencies for Gln223Arg or Pro1019Pro polymorphisms between obese and nonobese adolescents. However, there was a higher prevalence of Gln223 allele among subjects with higher insulin levels (0.72 vs 0.57; P = 0.04 for adolescents with insulin levels higher and lower than 100 pmol/l, respectively). According to Gln223Arg polymorphism, those with Gln allele (Gln/Gln and Gln/Arg) had higher heart sympathetic activity, body fat percentage, and leptin levels. To conclude, our results support the hypothesis that Gln223Arg polymorphism of LEPR in Mexican adolescents is associated with haemodynamic and metabolic disturbances related to obesity.

Pulmonary epithelial liquid absorption, expressed in relation to alveolar surface area, is reduced in fetal lambs following in utero tracheal occlusion.

We examined the effect of accelerated lung growth, induced by in utero tracheal occlusion (TO), on lung liquid uptake in near-term fetal sheep. In utero TO was performed in five fetal sheep at 110 days of gestation (term, approximately 145 days); six SHAM operated fetuses served as controls. The rate of liquid movement across the pulmonary epithelium was measured, using a previously established technique, in anesthetized fetal sheep between 133-137 days of gestation during a 2-hr adrenaline infusion (0.50 microg/min/kg, I.V.) and while lung luminal pressure was maintained at 5 mmHg. The rate of fetal lung liquid uptake was linear in all fetuses (mean r(2) < 0.97, n = 11). Mean values of lung liquid uptake expressed in relation to dry lung weight and luminal surface area of the right lung were significantly lower in TO fetuses (1.8 +/- 0.3 mL/hr/g and 1.0 +/- 0.2 mL/hr/m(2)) than in SHAM fetuses (2.6 +/- 0.2 mL/hr/g and 1.8 +/- 0.1 mL/hr/m(2)); surface area of the right lung was 140% greater in TO fetuses than in SHAM fetuses. There was a linear relationship between lung liquid uptake and pulmonary epithelial surface area in SHAM animals, but not in TO fetuses. We hypothesize that loss of alveolar epithelial type-II cells induced by increased levels of fetal lung expansion may impair alveolar liquid clearance in the perinatal period.

[Proposed normal values for alpha fetoprotein in maternal serum for the detection of neural tube closure defects and Down syndrome. Preliminary study]. / Propuesta de valores normales de alfa fetoproteína (AFP) en suero materno para la detección de defectos en el cierre del tubo neural (DCTN) y síndrome de Down (SD). Estudio preliminar.

To establish a normal range of alpha fetoprotein in maternal serum (AFP sm) in the population at the "20 de Noviembre" Hospital in Mexico City, there were studied 46 patients with a normal pregnancy confirmed with ultrasonography between 16 to 18 week of gestation. The 97 determinations of AFPsm were made by radioimmunoanalysis. The multiples of the median (MoM) were 2.16 al 16th weeks, 2.33 a 17th week 2.43 al 18th week of pregnancy. Now the methods to determine abnormalities in the product and the genetics studies are proposed only to the patients considered high risk (older than 35 years old, previous malformed son or family history of those abnormalities). AFPsm and the establishment of normal range allow us to amplify the study to every pregnant woman and to determine those malformations in the pregnant women of low risk which represents about 90-95% of DCTN and about 80% of SD.

Polyethylene wrap for thermoregulation in the preterm infant: a randomized trial.

We compared the response of temperature adaptation in preterm infants using the polyethylene wrap with and without previous drying versus the sterile preheated field. Both groups of polyethylene use achieved a mean axillary temperature of 36.5C at 30 minutes compared with 75 minutes for the group of traditional care. At 120 minutes, the incubator temperature was higher in those using preheated field, compared with infants in the polyethylene wrap with or without previous drying, (35.15C, 34.20C and 34.20C respectively; P = 0.0001). No difference in axillary or incubator temperature was found between the groups using the polyethylene wrap.

[Assessment of patients with distal radius fracture treated with percutaneous fixation]. / Evaluación de pacientes con fractura de radio distal tratados con fijación percutánea.

UNLABELLED: The trend in distal radius fracture is towards open reduction and internal fixation; however, treatment with closed reduction and percutaneous nailing continues to provide good results, but physical therapy and appropriate early rehabilitation are needed. MATERIAL AND METHODS: We assessed patients with distal radius fractures treated with reduction with percutaneous maneuvers and nailing; we used clinical functional scales, like the DASH, the Mayo Clinic wrist scale, and the Visual Analog Scale (VAS) to assess pain. Patient results were compared at 12 and 24 weeks. We also compared patients with and without rehabilitation. RESULTS: A total of 60 patients were assessed using the DASH scale; patients with rehabilitation had a score of 4.3, while those without rehabilitation had a score of 10.5 (p = 0.00001). In the Mayo Clinic scale, patients with rehabilitation had a score of 86.7, while those without rehabilitation had a score of 77.8 (p = 0.00001). The VAS was not significant. CONCLUSION: All patients showed improvement in their clinical assessment scales. However, when patients with and without rehabilitation are compared, the difference was greater in the Mayo Clinic scale and in the DASH scale.

Biochemical and molecular characterization of delta-endotoxins in Bacillus thuringiensis.

The delta-endotoxins (delta-ETX) of four native strains (RT7, RT19, RT25, and RT25), and one reference strain (4L1) of Bacillus thuringiensis were biochemically and molecularly characterized to determine their potential toxic activity against lepidopteran larvae. Crystals of delta-ETX were purified through a two-phase system to determine their morphology, molar mass, solubility, and resistance to proteinases. Toxic activity and cry gene content were also determined. Crystals from native strains exhibited polyhedral, irregular and cuboidal shapes, while those from 4L1 were bipyramidal. Seven proteins with estimated molar mass approximately 30-134 kDa were detected as the main components of the native delta-ETX. Only crystals from 4L1, RT24, and RT25 underwent complete solubilization at pH >12.0. Crystals from all strains produced trypsin-resistant peptides. None of the cry genes associated with toxicity in lepidopterans (cry1, cry2, cry9) was found in the native strains; however, 4L1 strain harbors cry1 and cry2 genes. Strains RT19 and RT25 caused significant mortality against Trichoplusia ni larvae with partial solubilization at pH 10, strain 4L1 caused 100 % mortality. Toxicity of native strains may come from a novel cry gene.

Effects of sterile high pressure vesicoureteral reflux on the monkey.

We surgically produced high-grade sterile vesicoureteral reflux in infant monkeys by unroofing the intravesical ureter and partially ligating the bladder neck. Half of the monkeys developed a decrease in renal function and were found to have interstitial nephritis. We found that this was due to a high resting bladder pressure, which affected the function and structure of the ureter. Thus, when a functional obstruction occurs in the refluxing ureter, renal function may be lost even when the urine is sterile. This situation is like that occurring in the patient with posterior urethral valves.

[Parameters for the evaluation of perinatal well-being: fetal and placental weight, placental and fetal ratio]. / Parámetros para evaluar el bienestar perinatal: peso fetal y placentario, cociente placentario y fetal.

Authors study fetal and placental weight during last weeks of gestation, as well as their correlations. At the end of gestation fetus grows more than placenta until the 40th week. From this moment, fetal growth slows and the placenta enlarges in order to maintain fetal homeostasis. Both ratios between placental and fetal weight are useful in evaluating parameters to show alterations of fetal and placental growth.

[Metabolism and elimination of ethosuximide in neonates whose mothers had petit mal]. / Metabolismo y eliminación de la etosuximida en el recién nacido de madre afecta de "petit mal".

In thirteen newborn infants of mothers affected with "petit mal" and treated with ethosuximide, submitted to artificial feeding with powdered milk, it was found that transplacental elimination of the drug occurred. Authors recorded daily serum concentration of ethosuximide and found that average life before it was totally eliminated was 80.62 +/- 7.24 hours. At the moment of birth serum concentration of mothers, and their newborn infants are similar due to transplacental by-pass. Through a lineal regression coefficient they find a relation between serum concentration in newborn infants and time passed from the moment of birth. This coefficient was negative and thus signified facility of the newborn to eliminate the drug. This result allows us to treat the mother adequately during anti-natal period with adequate dose to avoid subsequent epileptic attacks without producing toxicity in the newborn child.

Naltrexone does not prevent the weight gain and hyperphagia induced by the antipsychotic drug sulpiride in rats.

Few pharmacological tools are currently available to counteract the excessive body weight gain often observed during prolonged administration of antipsychotic drugs. Most antipsychotic drugs block dopamine receptors, and both the brain dopaminergic and opioid systems appear to be involved in initiation and maintenance of feeding behavior, respectively. We evaluated whether the opioid antagonist naltrexone (NAL, 0.5-16 mg/kg/ip for 21 days) (a) affects body weight and food intake in gonadally-intact and drug-free female rats, (b) prevents obesity, hyperphagia, hyperprolactinemia and vaginal cycle disruption induced by long-term administration of the antipsychotic drug sulpiride (SUL, 20 mg/kg/ip for 21 days), or (c) reverses the acute hyperphagia induced by SUL (15 microg bilaterally), when directly applied in the perifornical lateral hypothalamus (PFLH). In drug-free rats, only NAL doses above 4 mg/kg, significantly decreased weight gain and food intake. Even though NAL (1 and 8 mg/kg) significantly attenuated SUL-induced hyperphagia and hyperprolactinemia, it did not reverse at any dose the weight gain and permanent diestrous induced by SUL. In addition, local NAL did not prevent the hyperphagia and polidypsia observed after acute intrahypothalamic SUL. Unexpectedly, the cumulative and 24 h food intake in SUL-treated rats was significantly increased by NAL. Collectively, these results do not support a role for endogenous opiates in the neural and endocrine mechanisms involved in weight gain during prolonged antipsychotic drug administration in rats.

El interferon. / The interferon

Con el descubrimiento de los antibioticos, la medicina ha alcanzado un importante triunfo en su larga lucha contra las infecciones bacterianas, pero quedo rezagada en cuanto a las de origen viral. Sin embargo, el organismo no ha quedado desprotegido, ya que cuenta con una defensa natural contra los virus: EL INTERFERON. El descubrirlo, conocerlo y producirlo, ha sido y es objeto de ardua labor de investigacion, en la cual se ha invertido material humano, monetario y anos de intenso trabajo y estudio. De aqui, que el interferon ha rebasado en la actualidad los limites del campo de la virologia, invadiendo otras areas biomedicas, tales como la genetica, la inmunologia, la biologia molecular y la oncologia. El objetivo primordial de esta revision, es el actualizar los conocimientos generales que se tienen acerca del interferon