DNA Methylation Remodeling after Bariatric Surgery Correlates with Clinical Parameters.

The altered functions of adipose tissue are one of the main issues in obesity. Bariatric surgery is associated with improvement of obesity associated comorbidities. Here DNA methylation remodeling in adipose tissue after bariatric surgery is examined. After six months postoperative, DNA methylation shows changes in 1155 CpG sites, 66 of these sites correlate with body mass index. Some sites also show correlation with LDL-C, HDL-C, total cholesterol, and triglycerides. CpG sites are located in genes that have not previously been linked to obesity or metabolic diseases. GNAS complex locus is one of those that presented CpG site with the greatest changes after surgery, and the most significant correlation with BMI and lipid profiles. These results show that epigenetic regulation may be involved in the alteration of adipose tissue functions in obesity.

DNA methylation and gene expression analysis in adipose tissue to identify new loci associated with T2D development in obesity.

BACKGROUND: Obesity is accompanied by excess adipose fat storage, which may lead to adipose dysfunction, insulin resistance, and type 2 diabetes (T2D). Currently, the tendency to develop T2D in obesity cannot be explained by genetic variation alone-epigenetic mechanisms, such as DNA methylation, might be involved. Here, we aimed to identify changes in DNA methylation and gene expression in visceral adipose tissue (VAT) that might underlie T2D susceptibility in patients with obesity. METHODS: We investigated DNA methylation and gene expression in VAT biopsies from 19 women with obesity, without (OND = 9) or with T2D (OD = 10). Differences in genome-scale methylation (differentially methylated CpGs [DMCs], false discovery rate < 0.05; and differentially methylated regions [DMRs], p value < 0.05) and gene expression (DEGs, p value <0.05) between groups were assessed. We searched for overlap between altered methylation and expression and the impact of altered DNA methylation on gene expression, using bootstrap Pearson correlation. The relationship of altered DNA methylation to T2D-related traits was also tested. RESULTS: We identified 11 120 DMCs and 96 DMRs distributed across all chromosomes, with the greatest density of epigenomic alterations at the MHC locus. These alterations were found in newly and previously T2D-related genes. Several of these findings were supported by validation and extended multi-ethnic analyses. Of 252 DEGs in the OD group, 68 genes contained DMCs (n = 88), of which 24 demonstrated a significant relationship between gene expression and methylation (p values <0.05). Of these, 16, including ATP11A, LPL and EHD2 also showed a significant correlation with fasting glucose and HbA1c levels. CONCLUSIONS: Our results revealed novel candidate genes related to T2D pathogenesis in obesity. These genes show perturbations in DNA methylation and expression profiles in patients with obesity and diabetes. Methylation profiles were able to discriminate OND from OD individuals; DNA methylation is thus a potential biomarker.

Programa cubano de pesquisa neonatal de la hiperplasia suprarrenal congénita: una realidad. 2005-2014 / Cuban neonatal screening program for congenital adrenal hyperplasia: a reality. 2005­2014

Introducción: la hiperplasia suprarrenal congénita (HSC) es el desorden adrenal más común en la infancia y la causa más frecuente de ambigüedad sexual. La forma clásica, que representa los casos más severos de este déficit, se asocia en un 75 % con pérdida de sal. Por otra parte, en los recién nacidos (RN) del sexo femenino se pueden presentar grados severos de virilización de genitales. Objetivo: presentar los resultados (durante diez años), del Programa Cubano de Pesquisa Neonatal (PN) de la HSC, soportado en un procedimiento inmunoenzimático desarrollado en Cuba. Resultados: en el período de enero 2005 a diciembre 2014, se han estudiado 1 140 882 RN y se detectaron 56 niños con HSC, para una incidencia de 1:20 373 RN. La cobertura del programa se ha incrementado hasta llegar en el año 2013 al 99.34 % de todos los RN cubanos. Conclusiones: la existencia del Programa Cubano de PN de HSC, ha permitido estimar la incidencia e incrementar el conocimiento de esta enfermedad. La PN ha posibilitado el diagnóstico precoz en la variedad perdedora de sal, contribuyendo a la disminución de la mortalidad infantil. El Programa ha favorecido a pacientes con formas virilizantes de la enfermedad, mediante la asignación correcta del sexo

Introduction: Congenital Adrenal Hyperplasia (HSC) is the most common adrenal disorder in childhood and the most frequent cause of sexual ambiguity. The classic form, which represents the most severe causes of this deficit, is associated in 75 % with loss of salt. On the other hand, in the NB of the female sex that present severe degrees of virilization of the genitals. Objective: To present result of the application for ten years of the Cuban Neonatal Research Program of the HSC, supported by an inmmunoenzymatic procedure developed in Cuba. Result: In the period from January 2005 to December 2014, using the UMELISA 17 OH PROGESTERONA NEONATAL, 1 140882 RN were detected, for an incidence of 1:20373 RN. The coverage of the program has been increasing until2013 reaching 99.34 % of all Cuban RN. Conclusion: The existence of the Cuban HSC PN Program, has allowed estimating the incidence and increase knowledge of this disease in our country. PN has made possible the early diagnosis of patients with the salt losing variety, contributing to decrease in infant mortality nationwide. The Program has favored patients with virilizing form of the disease, through the correct assignment of sex

The interplay between internal and external load parameters during different strength training sessions in resistance-trained men.

External load influences internal load in resistance training (RT). The purpose of the present study was to compare the total volume-load, perceptual and stress responses during three different RT protocols. Twelve resistance-trained men completed three different RT protocols with the back squat and bench press exercises: (1) power (POW) (5 sets of 6 repetitions at 50%1RM, 2-min of rest), (2) hypertrophy (HYP) (5 sets-to-failure at 75%1RM, 2-min of rest), and (3) strength (STR) (5 sets-to-failure at 90%1RM, 3-min of rest). Volume-load (kg × reps.), session rating of perceived exertion (sRPE), training impulse (TRIMP; reps. × sRPE), cortisol, immunoglobulin A (IgA), lactate, and creatine kinase (CK) were assessed before and/or after the sessions. HYP was the most demanding session in terms of volume-load (p < 0.001), TRIMP (p < 0.001), cortisol (p = 0.001), lactate (p < 0.001), and CK (p = 0.001). Despite POW exhibited a greater volume-load than STR (p = 0.016), the latter exhibiting a greater sRPE (p < 0.001), and a greater post-session CK (p = 0.05). However, the TRIMP of STR and POW were not statistically different (152 vs. 260 AU; p = 0.089). These specific responses could be meditated by the presence of muscular failure. When pooling all the sessions, significant correlations were revealed among external and internal stress markers (r = 0.35-0.80; p ≤ 0.05). The use of TRIMP could be recommended as a simple and valid monitoring tool which integrates into a single parameter the internal and the external loads of RT sessions.

Pilot study for cystic fibrosis neonatal screening: the Cuban experience.

Background In Cuba, no screening program for cystic fibrosis (CF) has been implemented yet. The ultramicro enzyme-linked immunosorbent assay (UMELISA)® TIR NEONATAL has been developed for the measurement of immunoreactive trypsin (IRT) in dried blood spots on filter paper. The analytical performance of the kit was evaluated in the national network of laboratories. Methods Newborn dried blood samples (DBS) were evaluated in 16 laboratories. An IRT/IRT/DNA protocol was followed using a cut-off value of 50 ng/mL. The mean, median and percentiles of the distribution were calculated and a two-sample t-test with unequal variance was used for statistical analysis. Influence of perinatal factors on IRT levels was analyzed. Results From January to June 2018, 6470 newborns were studied, obtaining a mean IRT value of 12.09 ng/mL (ranging 0-358 ng/mL) and a median of 8.99 ng/mL. Fifty-two samples (0.78%) were above the cut-off level and 16 samples (0.24%) were elevated in the re-screening process. One of them was confirmed positive by molecular biology (phe508del/c.3120 + 1G > A), constituting the first newborn screened and diagnosed early in Cuba. Second DBS samples were collected on average at 14 days and processed in the laboratory at 16 days of birth. Significant differences were observed (p < 0.05) when evaluating the influence of gender, birth weight (BW) and gestational age (GA) on the IRT values. Lower IRT concentrations were found in samples processed after 10 days of collection. Conclusions The performance of UMELISA® TIR NEONATAL in the laboratories has been satisfactory; hence CF newborn screening (NBS) was extended throughout the country from January 2019.

Is Perceived Exertion a Useful Indicator of the Metabolic and Cardiovascular Responses to a Metabolic Conditioning Session of Functional Fitness?

The purpose of this study was to assess whether the self-regulation of training intensity based on rating of perceived exertion (RPE) is a reliable method to control the intensity during metabolic conditioning sessions of functional fitness. In addition, the relationship between RPE and the changes in heart rate, number of repetitions, and lactate responses was also analyzed. Eight male participants (age 28.1 ± 5.4 years; body mass 77.2 ± 4.4 kg; VO2 max: 52.6 ± 4.6 mL·(kg·min)-1 completed two sessions (five to seven days apart), in a randomized order, under different conditions, as follows: (1) all-out (ALL), or (2) self-regulation of intensity based on an RPE of six (hard) on the Borg CR-10 scale (RPE6). The rating of perceived exertion, lactate (LAC), and heart rate (HR) response were measured before, during, and immediately after the sessions. The RPE and LAC during the all-out sessions were higher (p < 0.0005) than the RPE6 session for all of the analyzed time points during the session. There was no difference in the HR area under the curve for the all-out and RPE6 sessions. The average number of repetitions performed was lower (p ≤ 0.009) for the RPE6 session (190.5 ± 12.5 repetitions) when compared to the all-out session (214.4 ± 18.6 repetitions). There was a significant correlation between the RPE and LAC (p = 0.005; r = 0.66; large) and number of repetitions during the session (p = 0.026; r = 0.55; large). No correlation was observed between the RPE and HR (p = 0.147; r = 0.380). These results indicate that the self-regulation of intensity of effort based on the RPE may be a useful tool to control the exercise intensity during a metabolic conditioning session of functional fitness.

Vertical Jump Is Strongly Associated to Running-Based Anaerobic Sprint Test in Teenage Futsal Male Athletes.

As one of the most popular sport modalities in Brazil, and with an exponential growth in Europe, futsal is characterized by intermittent stimulus of anaerobic high intensity sprints. The running-based anaerobic sprint test (RAST) is one of the most common tests to assess anaerobic power in futsal athletes, however, it presents both time and physical challenges. Therefore, we aimed to correlate RAST with a simpler test, the vertical jump (VJ), in teenage male futsal athletes; Methods: Thirteen volunteers were enrolled and underwent two visits to the laboratory, one for the VJ and the other for the RAST in a randomized order; Results: The association test indicates a strong and significant correlation between VJ and RAST. We conclude that VJ can be used as an alternative to RAST in teenage male futsal athletes.

An Enzyme Immunoassay for Determining Immunoreactive Trypsinogen (IRT) in Dried Blood Spots on Filter Paper Using an Ultra-Microanalytical System.

Cystic fibrosis (CF) is a severe autosomal recessive disorder. It is caused by mutations in the CF transmembrane conductance regulator gene. Early diagnosis of CF can be carried out by determining high immunoreactive trypsinogen (IRT) blood values in newborns. A simple sandwich-type ultramicroELISA assay (UMELISA®) has been developed for the measurement of IRT in dried blood spots on filter paper. Strips coated with a high affinity monoclonal antibody directed against IRT are used as solid phase, to ensure the specificity of the assay. The assay is carried out within 20 h. The useful rank of the curve is 0-500 ng/mL, and the lowest detectable concentration is 4.8 ng/mL. Intra- and inter-assay coefficients of variation were lower than 10%. The recovery mean value was 100.3 ± 11.2%. Cross-reactivity with proteins structurally related to IRT (α2-macroglobulin, α1-antitrypsin, and human chymotrypsin) was lower than the detection limit of the assay. Four thousand four hundred six newborn samples from the Cuban Newborn Screening Program were analyzed, and the mean IRT concentration was 12.8 ng/mL. Higher IRT values were obtained when samples were eluted overnight. Regression analysis showed a good correlation with the commercially available AutoDELFIA® Neonatal IRT kit (n = 3948, r = 0.885, ƙ = 0.976, p < 0.01). The analytical performance characteristics of our UMELISA® TIR Neonatal suggest that it can be used for the neonatal screening of CF.

An enzyme immunoassay for determining epidermal growth factor (EGF) in human serum samples using an ultramicroanalytical system.

Human epidermal growth factor is a small peptide consisting of 53 amino acid residues, which stimulates cell proliferation and is associated with several human carcinomas. A simple sandwich-type ultramicroELISA assay (UMELISA), based on the advantages of high affinity reaction between streptavidin and biotin has been developed for the measurement of EGF in human serum samples. Strips coated with a high affinity monoclonal antibody directed against EGF are used as solid phase, to ensure the specificity of the assay. The EGF assay was completed in 18 hr, with a measuring range of 39-2500 pg/mL. The intra- and inter-assay coefficients of variation were 4.4-7.3% and 0-5.1%, respectively, depending on the EGF concentrations evaluated. Percentage recovery ranged from 96-104%. Regression analysis showed a good correlation with the commercially available Human EGF Immunoassay Quantikine® ELISA kit (n = 130, r = 0.92, P < 0.01). The analytical performance characteristics of our UMELISA EGF endorse its use for the quantification of EGF in human serum samples.

Contact Karate Promotes Post-Exercise Hypotension in Young Adult Males.

BACKGROUND: Worldwide, systemic arterial hypertension is a leading cause of death and non-communicable cardiovascular disease. A major factor contributing to this disease is a sedentary lifestyle. However, physical exercise, such as martial arts, may be an option for blood pressure (BP) control. The magnitude of post-exercise hypotension is associated with a prolonged decrease in BP in normotensive and hypertensive individuals. OBJECTIVES: The present study aimed to verify the effects of a Contact Karate (CK) session on BP responses during a post-exercise recovery period in young adults. PATIENTS AND METHODS: Thirty-two male CK athletes volunteered (28.2 ± 6.7 years; 77.0 ± 5.7 kg; and 176.0 ± 4.7 cm) and underwent one CK session (50 minutes) and a control session in which no exercise was performed and the individuals remain seated during the whole time. BP was measured during rest (before sessions), as well as on the 15th, 30th, 45th, and 60th minutes of the post-exercise recovery. RESULTS: The systolic (SBP), diastolic (DBP), and mean arterial pressure (MAP) were significantly lower at the post-exercise period compared to pre-exercise rest (P < 0.05), with the largest reductions being observed at the 60th minutes of recovery [SBP (rest: 125.9 ± 4.7 vs. 60th minutes of recovery: 111.7 ± 5.4 mmHg); DBP (rest: 78.8 ± .7 vs. 60th minutes of recovery: 69.8 ± 2.7 mmHg)] and at the same periods of post-exercise recovery of the control session. CONCLUSIONS: A single CK session can promote a decrease in BP for at least 60 minutes after performing this type of exercise in young adults.

SUMA Technology and Newborn Screening Tests for Inherited Metabolic Diseases in Cuba: An Overview of the First 30 Years

Abstract The ultramicroanalytic system (SUMA), created in the 1980s, is a complete system of reagents and instrumentation to perform ultramicroassays combining the sensitivity of the micro-enzyme-linked immunosorbent assay (ELISA) tests with the use of ultramicrovolumes. This technology permitted establishing large-scale newborn screening programs (NSPs) for metabolic and endocrine disorders in Cuba. This article summarizes the main results of the implementation during the 30 years of SUMA technology in NSP for 5 inherited metabolic diseases, using ultramicroassays developed at the Department of Newborn Screening at the Immunoassay Center. Since 1986, SUMA technology has been used in the Cuban NSP for congenital hypothyroidism, initially studying thyroid hormone in cord serum samples. In 2000, a decentralized program for the detection of hyperphenylalaninemias using heel dried blood samples was initiated. These successful experiences permitted including protocols for screening congenital adrenal hyperplasia, galactosemia, and biotinidase deficiency in 2005. A program for the newborn screening of CH using the thyroid-stimulating hormone Neonatal ultramicro-ELISA was fully implemented in 2010. Nowadays, the NSP is supported by a network of 175 SUMA laboratories. After 30 years, more than 3.8 million Cuban newborns have been screened, and 1002 affected children have been detected. Moreover, SUMA technology has been presented in Latin America for over 2 decades and has contributed to screen around 17 million newborns. These results prove that developing countries can develop appropriate diagnostic technologies for making health care accessible to all.

Exercise performed around MLSS decreases systolic blood pressure and increases aerobic fitness in hypertensive rats.

BACKGROUND: Exercise is a non-pharmacologic agent widely used for hypertension control, where low intensity is often associated with blood pressure reduction. Maximal lactate steady state (MLSS) was recently identified in spontaneously hypertensive rats (SHRs) as an important step in establishing secure intensities for prescribing exercise for hypertensive phenotypes. Here we verified the effects of training around MLSS, 20% below MLSS, and 15% above MLSS on aerobic fitness and blood pressure status of SHR. Eighteen-week-old SHRs (n = 5, ~ 172.4 ± 8.1 mm Hg systolic blood pressure) were trained on a treadmill for 4 weeks for 30 min/day, 5 days/week at a velocity of 20 m.min(-1). After training, a novel MLSS and incremental test was performed to evaluate the animals' aerobic fitness. Furthermore, ~ 22-week-old SHRs (n = 12, ~169.8 ± 13.8 mm Hg systolic blood pressure) were divided into non-exercised (CG, n = 4), low intensity (LIG, n = 4) and high intensity (HIG, n = 4) groups, where rats were trained at 16 m.min(-1) and 23 m.min(-1) respectively for 30 min/day, 5 days/week for 4 weeks. RESULTS: Exercise performed at MLSS enhanced aerobic fitness, leading to a novel MLSS, identified around 30 m.min(-1). Low and high intensity training reduced systolic blood pressure and only high intensity training led to improved aerobic fitness (28.1%, p < 0.01). CONCLUSIONS: Therefore, our data indicate a decrease in blood pressure due to low and high exercise intensity, and an increase in aerobic fitness provided by high-intensity exercise in SHRs.

Clinical, endoscopic and manometric features of the primary motor disorders of the esophagus.

BACKGROUND: Significant incidence, diagnostic difficulties, clinical relevance and therapeutic efficacy associated with the small number of publications on the primary esophageal motor disorders, motivated the present study. AIM: To determine the manometric prevalence of these disorders and correlate them to the endoscopic and clinical findings. METHODS: A retrospective study of 2614 patients, being 1529 (58.49%) women and 1085 (41.51%) men. From 299 manometric examinations diagnosed with primary esophageal motor disorder, were sought-clinical data (heartburn, regurgitation, dysphagia, odynophagia, non-cardiac chest pain, pharyngeal globe and extra-esophageal symptoms) and/or endoscopic (hiatal hernia, erosive esophagitis, food waste) that motivated the performance of manometry. RESULTS: Were found 49 cases of achalasia, 73 diffuse spasm, 89 nutcracker esophagus, 82 ineffective esophageal motility, and six lower esophageal sphincter hypertension. In relation to the correlations, it was observed that in 119 patients clinical conditions were associated with dysphagia, found in achalasia more than in other conditions; in relationship between endoscopic findings and clinical conditions there was no statistical significance between data. CONCLUSION: The clinical and endoscopic findings have little value in the characterization of the primary motor disorders of the esophagus, showing even more the need for manometry, particularly in the preoperative period of gastroesophageal reflux disease.

Hipotermia postoperatoria causada por anestesia regional en pacientes sometidas a cesáreas electivas / Postoperative hypothermia caused by regional anesthesia in patients undergoing to elective cesarean section

Objetivo: Determinar la incidencia de hipotermia postoperatoria causada por anestesia regional en pacientes sometidas a cesáreas electivas en el Hospital Nacional Arzobispo Loayza en el periodo de Mayo a Julio del 2014. Material y método: se realizó un estudio observacional analítico, no experimental, sin grupo control y transversal en 305 pacientes sometidas a cesáreas electivas con anestesia regional en el Hospital Nacional Arzobispo Loayza en el periodo de mayo a julio del 2014. Los datos se procesaron según paquete estadístico SPSS v.21. Se utilizó el Chi cuadrado para determinar la significancia estadística y los intervalos de confianza, de acuerdo al análisis bivariante de las variables considerada. (P menor de 0.05 se considera significativo). Resultados: El tipo de anestesia regional más usada fue la epidural en un 57,7 por ciento de los casos. El 6,6 por ciento de las pacientes ya habían sido cesareadas y el 93,4 por ciento nunca habían sido cesareadas. La frecuencia de hipotermia no visible fue del 13,4 por ciento y de actividad muscular en un solo grupo fue del 12,1 por ciento de los casos respectivamente. En la mayoría de los casos que presentaron hipotermia, esta se presentó a los 60 minutos en el 18,7 por ciento de los casos. El 78,4 por ciento de los casos no presentaron dolor. La media global de la edad fue de 25,1+/-3,1 años siendo la mínima de 19 años y la máxima de 35 años. Hubo una mayor frecuencia de hipotermia en las pacientes en quienes se usó la anestesia epidural (55,4 por ciento), cesareadas primera vez (92,4 por ciento), presentándose con mayor frecuencia a los 60 minutos en el 62,0 por ciento de los casos. Conclusiones: La incidencia de hipotermia postoperatoria causada por anestesia regional en pacientes sometidas a cesáreas electivas en el Hospital Nacional Arzobispo Loayza fue del 30,2 por ciento. Hubo una diferencia estadísticamente significativa de tendencia a mayor edad en las pacientes que presentaron hipotermia (p<0.05)...

Clinical, endoscopic and manometric features of the primary motor disorders of the esophagus / Aspectos clínicos, manométricos e endoscópicos dos distúrbios motores primários do esôfago

BACKGROUND: Significant incidence, diagnostic difficulties, clinical relevance and therapeutic efficacy associated with the small number of publications on the primary esophageal motor disorders, motivated the present study. AIM: To determine the manometric prevalence of these disorders and correlate them to the endoscopic and clinical findings. METHODS: A retrospective study of 2614 patients, being 1529 (58.49%) women and 1085 (41.51%) men. From 299 manometric examinations diagnosed with primary esophageal motor disorder, were sought-clinical data (heartburn, regurgitation, dysphagia, odynophagia, non-cardiac chest pain, pharyngeal globe and extra-esophageal symptoms) and/or endoscopic (hiatal hernia, erosive esophagitis, food waste) that motivated the performance of manometry. RESULTS: Were found 49 cases of achalasia, 73 diffuse spasm, 89 nutcracker esophagus, 82 ineffective esophageal motility, and six lower esophageal sphincter hypertension. In relation to the correlations, it was observed that in 119 patients clinical conditions were associated with dysphagia, found in achalasia more than in other conditions; in relationship between endoscopic findings and clinical conditions there was no statistical significance between data. CONCLUSION: The clinical and endoscopic findings have little value in the characterization of the primary motor disorders of the esophagus, showing even more the need for manometry, particularly in the preoperative period of gastroesophageal reflux disease. .

RACIONAL: A incidência significante, a dificuldade diagnóstica, a relevância clínica e a eficácia terapêutica associada ao pequeno número de publicações sobre os distúrbios motores primários do esôfago, motivou a realização do presente estudo. OBJETIVO: Verificar a prevalência desses distúrbios em manometrias e relacioná-las aos achados endoscópicos e clínicos. MÉTODOS: Estudo retrospectivo de 2614 pacientes sendo 1529 (58,49%) do gênero feminino e 1085 (41,51%) do masculino. A partir de 299 exames manométricos com diagnóstico de distúrbio motor esofagiano primário, procuraram-se os dados clínicos (pirose, regurgitação, disfagia, odinofagia, dor torácica não cardíaca, globo faríngeo e sintomas extra-esofageanos) e/ou endoscópicos (hérnia de hiato, esofagite erosiva, resíduos alimentares) que motivaram a realização da manometria. RESULTADOS: Foram encontrados 49 casos de acalásia, 73 de espasmo difuso, 89 de esôfago em quebra-nozes, 82 de motilidade esofagiana ineficaz, e seis de esfíncter esofagiano inferior hipertensivo. Em relação às correlações, observou-se em 119 pacientes analisados que, na clínica associada às afecções, a disfagia foi mais encontrada na acalásia do que nas outras afecções; na relação entre os achados endoscópicos e as afecções não houve relevância estatística entre os dados. CONCLUSÃO: Os achados clínicos e endoscópicos têm pequeno valor na caracterização das doenças motoras primárias do esôfago, evidenciando ainda mais a necessidade da manometria, particularmente no pré-operatório da doença do refluxo gastroesofágico. .

Newborn screening for congenital adrenal hyperplasia in Cuba: six years of experience.

BACKGROUND: Since 2005, a newborn screening program for congenital adrenal hyperplasia (CAH) by measuring 17-alpha-hydroxyprogesterone (17OHP) in dried blood spots was introduced in Cuba. METHODS: The hormone was measured by the 17OHP Neonatal UMELISA method, in samples collected on the 5th day as average. Confirmatory test was performed to those neonates with 17OHP values above 55 nmol/l. Some perinatal factors that can influence on 17OHP levels were studied. RESULTS: From January 2005 to December 2010, 621,303 newborns were screened and 39 CAH cases were detected. Coverage of the program reached 98%. The incidence of CAH in Cuba was 1:15,931, similar to that reported by other programs. A recall for suspected CAH was performed in 10,799 cases (1.74%). Therapy in classical CAH patients was started at the mean age of 22 days. 17OHP levels were significantly higher in newborns with lower birth-weight (BW) and/or gestational age (GA). In addition, 17OHP values were affected by the gender, twin status or mode of delivery. CONCLUSIONS: In Cuba, the nationwide newborn screening program has allowed the early detection of CAH. The use of an optimized cut-off level for BW or GA could lead to a reduction in the percentage of recalled babies.

Noninvasive method to estimate anaerobic threshold in individuals with type 2 diabetes.

BACKGROUND: While several studies have identified the anaerobic threshold (AT) through the responses of blood lactate, ventilation and blood glucose others have suggested the response of the heart rate variability (HRV) as a method to identify the AT in young healthy individuals. However, the validity of HRV in estimating the lactate threshold (LT) and ventilatory threshold (VT) for individuals with type 2 diabetes (T2D) has not been investigated yet. AIM: To analyze the possibility of identifying the heart rate variability threshold (HRVT) by considering the responses of parasympathetic indicators during incremental exercise test in type 2 diabetics subjects (T2D) and non diabetics individuals (ND). METHODS: Nine T2D (55.6 ± 5.7 years, 83.4 ± 26.6 kg, 30.9 ± 5.2 kg.m2(-1)) and ten ND (50.8 ± 5.1 years, 76.2 ± 14.3 kg, 26.5 ± 3.8 kg.m2(-1)) underwent to an incremental exercise test (IT) on a cycle ergometer. Heart rate (HR), rate of perceived exertion (RPE), blood lactate and expired gas concentrations were measured at the end of each stage. HRVT was identified through the responses of root mean square successive difference between adjacent R-R intervals (RMSSD) and standard deviation of instantaneous beat-to-beat R-R interval variability (SD1) by considering the last 60 s of each incremental stage, and were known as HRVT by RMSSD and SD1 (HRVT-RMSSD and HRVT-SD1), respectively. RESULTS: No differences were observed within groups for the exercise intensities corresponding to LT, VT, HRVT-RMSSD and HHVT-SD1. Furthermore, a strong relationship were verified among the studied parameters both for T2D (r = 0.68 to 0.87) and ND (r = 0.91 to 0.98) and the Bland & Altman technique confirmed the agreement among them. CONCLUSION: The HRVT identification by the proposed autonomic indicators (SD1 and RMSSD) were demonstrated to be valid to estimate the LT and VT for both T2D and ND.

Quantitative ultramicrotest for newborn screening of galactosemia in Cuba.

BACKGROUND: To describe a simple, rapid, quantitative ultramicrotest (UMTEST) based on the fluorometric method introduced by Fujimura et al. adapted to an Ultra Micro Analytic System (SUMA) for the detection of total galactose (GAL) in dried blood specimens. METHODS: The assay uses 3 mm discs of dried blood on Whatman 903 filter paper and small volumes of each reagent. A methanol/acetone/water solution is used for deproteination, and a specially designed 96-well polystyrene opaque ultramicroplates, with a maximum capacity of 30 µL per well, are used for the reading. RESULTS: The UMTEST GAL is completed in 2 h, with measuring range of 0.28-3.92 mmol/L. The intra- and inter-assay coefficients of variation were 2.3%-8.9% and 6.8%-11.1%, respectively, depending on the total GAL concentrations. Percentage recovery ranged from 97.7% to 103%. Limit of detection and limit of quantitation were 0.06 and 0.16 mmol/L, respectively. The mean GAL concentration, in 2510 dried blood samples from the National Neonatal Screening Program was 0.23 mmol/L. Our assay showed high concordance correlations with the commercially available ICN Immuno-Chem™ GAL-MW EA kit. CONCLUSIONS: The analytical performance characteristics of this assay is suitable for mass newborn screening of galactosemia in Cuba.

Cuban neonatal screening of phenylketonuria using an ultramicro-fluorometric test.

BACKGROUND: Guthrie's bacterial inhibition assay was used in Cuba, since 1983. A decentralized program for the newborn screening of hyperphenylalaninemias started in the year 2000 using an ultramicro-fluorometric test (UMTEST PKU). METHODS: A simple and rapid ultramicro-fluorometric test based on McCaman and Robin's method has been designed, developed and applied for the measurement of Phe in dried blood spots on filter paper. RESULTS: The UMTEST PKU exhibited an acceptable precision and accuracy. Samples of 27528 newborns on filter paper Schleicher & Schuell 903 (S&S 903) from the National neonatal screening program were collected and analyzed, and the mean Phe concentration was 66.5 micromol/l. Our assay showed high Pearson and concordance correlations with 2 commercially available kits. A total of 521923 Cuban newborns were studied from the year 2000 to 2007 using the UMTEST PKU. Elevated blood phenylalanine levels were found in 1764 infants (0.34%) and no false negative were noted. Ten cases were diagnosed with phenylketonuria, all of them with an initial phenylalanine concentration over 360 micromol/l. CONCLUSIONS: The analytical performance characteristics of our assay and its use in the National program have demonstrated its suitability for the neonatal screening of PKU.