Evaluation of reference genes for expression studies in leukocytes from term human pregnancy.

INTRODUCTION: Human labor is considered an inflammatory process modulated by systemic and local leukocytes that infiltrate into the maternal-fetal interface. The putative roles of these leukocytes are currently being studied with gene expression assays. Such assays are normalized by the expression of housekeeping genes. However, expression of housekeeping genes may vary depending on the cell type and/or the experimental conditions. The aim of this study was to analyze the expression stability of several housekeeping genes in leukocytes from term human pregnancies, considering both anatomical origin and presence of labor. METHODS: We analyzed the gene expression of ACTB, B2M, GAPDH, GUSB, PGK1, RN18S1, TBP and UBC in leukocytes from maternal peripheral blood, placental blood and choriodecidua in women delivering at term with or without the presence of labor through real-time qPCR. Then we used geNorm to evaluate expression stability and pairwise variation. RESULTS: The expression of all tested genes showed to be stable independent of the anatomical compartment and the absence or presence of labor. However, PGK1, GUSB and TBP showed to be the most stable and RN18S1 the least stable. Pairwise variation analyses showed that only two genes are needed for normalization yet the inclusion of a third improves its accuracy. DISCUSSION: PGK1, GUSB and TBP are the most adequate reference genes for gene expression normalization in leukocytes from term pregnancies, regardless of their anatomical origin (maternal peripheral blood, placental blood or choriodecidua) or the presence or absence of labor. Our study is the first report on housekeeping gene stability in leukocytes from healthy pregnant women.

Human Immunodeficiency Virus type 1 in seronegative infants born to HIV-1-infected mothers.

BACKGROUND: Some individuals repeatedly exposed to Human Immunodeficiency Virus do not seroconvert and are resistant to HIV infection. Here, in a pediatric cohort of HIV seronegative infants born of HIV-infected mothers, we have studied eight non-breastfed children in whom viral DNA was detected in their PBMC. Our objective was to assess whether silent infection in these children can be explained by the presence of integrated viral DNA. METHODS: The presence of viral DNA was corroborated by nested PCR with primers for gag and the nef/LTR regions of HIV-1. Integration of HIV DNA into the host genome was assessed by an Alu-LTR PCR. Amplicons were sequenced and phylogenetic analyzes were done. RESULTS: HIV-1 DNA was detected in the earliest available PBMC sample from all eight infants, and two of them tested positive for HIV DNA at 2 years of age. Nested PCR resulted in the amplification of gag, nef/LTR and Alu-LTR fragments, which demostrated that HIV-1 DNA was integrated in the host cell genome. Each individual has a characteristic sequence pattern and is different from the LTR sequence of HXB2 prototype virus and other Mexican isolates. CONCLUSION: HIV-1 DNA was observed in PBMC from HIV exposed seronegative children in this pediatric cohort.

[Isolated hypoplasia of the right ventricle with interatrial communication. Study of a case and review of the literature]. / Hipoplasia aislada de ventrículo derecho con comunicación interauricular. Estudio de un caso y revisión de la literatura.

A 1 1/2 year old boy with cyanosis and congestive heart failure was found to have isolated right ventricular hypoplasia with atrial septal defect by cardiac catheterization and angiocardiography. On physical examination no murmur was heard. The ECG revealed a frontal plane AQRS of -45 degrees, right atrial enlargement and left ventricular hypertrophy. Chest X-ray examination showed slight cardiomegaly. Preoperative assessment included temporary occlusion of the atrial septal defect by means of a balloon catheter. Surgical closure of the atrial septal defect was performed at 4 years old. The clinical findings and surgical treatment of this rare malformation are discussed.

[Growth in 60 children with surgically repaired interventricular communication]. / Crecimiento de 60 niños con comunicación interventricular operada.

Growth status was studied in 60 children with simple and severe ventricular septal defect successfully repaired. The average follow-up post-surgery was 3,5 years. Children with birth weight less of 2.500 g and extracardiac anomalies or chromosomal syndromes were excluded. Measurements before surgical treatment revealed height and weight status to be subnormal and related to physiological severity. However lowest weight score was found in youngest patients and height score was uniformly affected in both group ages. Surgical repair resulted in significant weight gain in all groups. Final height in the youngest age groups is near normal levels. However in the group of children repaired after 2 years age of the changes in height were not statistically significant. No change in the subnormal growth pattern was noted in 6 children (10%). Surgery of large and severe ventricular septal defect in very early infancy offers the best prospect for growth.

[Growth in 60 children with surgically corrected tetralogy of Fallot]. / Crecimiento de 60 niños con tetralogía de Fallot operada.

Growth status was followed in 60 children with Fallot's tetralogy for a mean of 3,3 years after total correction. In 32 of the patients a palliative shunt had been performed earlier with no effect on their growth. Complete correction improved growth status in 50 children. Best results were seen when the correction was performed between 2 and 4 years of age. Ten children (16.6%) did not improve their growth status; 5 of them underwent surgical correction before 2 years of age, although only 11 of the 50 children with significant improvement were operated before this age. Pathophysiological and therapeutic implications of these findings are discussed.

[Growth in 48 children with ventricular septal defect successfully repaired (author's transl)]. / Crecimiento de 48 niños con comunicación interventricular operada.

Growth status was studied in 48 children with simple and severe ventricular septal defect successfully repaired. The average follow-up post surgery was 3.5 years (range, two-seven years). Children with birth weight less of 2,500 g. and extracardiac anomalies or chromosomal syndromes were excluded. Measurements before surgical treatment revealed height and weight status to be subnormal and related to physiological severity. However lowest weight score was found in youngest patients and height score are uniformly affected in both group ages. Surgical repair resulted in significant weight gain in all groups. Final height in the youngest age groups is near normal levels. However in the group of children repaired after age of two years the changes in height were not statistically significant. No change in the subnormal growth pattern was noted in five children (10.4%). Surgery of large and severe ventricular septal defect in very early infancy offers the best prospect for growth.

[Interventricular communication with infundibular reaction (author's transl)]. / Comunicación interventricular con reacción infundibular.

Ten patients exhibiting an interventricular communication and an infundibular reaction are presented. As a key premise for selection of cases, an angiohemodynamic study and presence of a gradient at an infundibular level above 25 mm. Hg. were required. Clinic characteristics as well as electrocardiogram, radiology and angiohemodynamic study of all patients are commented. Incidence in different studies, as well as in our own, is discussed, which was found to be of 9%. Similarly, diagnosis of evolutive form of interventricular communication and evolutive character in question is commented; four cases of their own are provided. The various explanatory theories of infundibular stenosis are commented.

[Pulmonary atresia with intact ventricular septum (author's transl)]. / Obstrucción severa del tracto de salida del ventrículo derecho. I. Atresia pulmonar con "septum" interventricular íntegro.

Ten cases of pulmonary atresia with intact ventricular septum are presented. The groups in which these patients are classified are described, thinking that the mentioned classification is a simplification of the problem. After discussion of the dates apported by clinic, thorax X-Rays and electrocardiogram, authors come to the conclusion that they are not specific of this cardiopathy. Contribution of hemodinamic and angiocardiographic characteristics, paying special attention to the blood flow to lung vascular tree and in existance of intramyocardium sinusoid are shown. Description of the general tendencies of the most accepted surgical techniques in bibliography for resolution of both groups of patients is made.

[Critical pulmonary stenosis in infants (author's transl)]. / Obstrucción severa del tracto de salida del ventrículo derecho.

Eighteen infants carrying pulmonary stenosis with a complete interventricular wall whose most frequent clinical data are: precocius cyanosis, congestive cardiac insufficiency and in 25% of them hypoxic crises are presented. Complementary explorations of this entity are not definitive, but clinical diagnosis described with evolutional cardiomegaly at the expense of the right cavities and pulmonary ischemia forces authors to think that there is a severe obstruction of the infumdibulum with complete interventricular septum. The different parameters which have been proposed to evaluate severity of the stenosis are discussed and compared, finding among them significative differences. Authors consider the concept of the right diminutive ventricle, finding in these cases the electrocardiographic pattern rS in V1 and observing a great mortality both spontaneous and postsurgical. The high mortality of this heart disease, both in its' natural evolution as well as after surgery is to be pointed out as in most published series.

[Toxic epidermal necrolysis (Lyell's syndrome) (author's transl)]. / Necrolisis epidérmica tóxica. Síndrome de Lyell.

One case of toxic epidermal necrolysis (Lyell's syndrome) in a 6 1/2 year old girl is presented. The patient had received penicillin and streptomycin for four days two weeks before the onset, and was admitted to the hospital with epidermal lesions characteristic of the bullous stage of Lyell's disease. A biopsy of the skin confirmed the diagnosis. Cultures of blood, skin and exudate from the bullae were negative, and immunological tests were normal. No immunohistologic studies on the skin biopsy were done. The patient was treated with cephalosporins and corticosteroids, and the evolution was favorable. The main clinical traits found in this patient are discussed on the light of information available from previous reports. Several aspects of the diagnosis, management, and natural history of the disease are also discussed.

[Atrial septal defect with pulmonary hypertension in infancy (author's transl)]. / Comunicación interauicular con hipertensión pulmonar en el lactante.

Four cases of atrial septal defect, "ostium secundum", associated with pulmonary hypertension and congestive heart failure in children under the age of two years are reported. A correct diagnosis could be made before surgery in only two of the case. In one case partial anomalous pulmonary venous connection was incorrectly diagnosed, while ventricular septal defect could not be excluded in the fourth case. The systolic pressure in the pulmonary artery was higher than 50 mm. Hg in all cases. One patient died after catheterization, and total surgical correction was performed in the other three cases, with good results. In all of them, a large atrial septal defect could be seen. There was no evidence of any other associated anomaly. After an average follow up of 4.5 years the three patients that where operated remain asymptomatic. The rarity of this entity, the difficulties in making a correct diagnosis and several pathogenetic aspects of this abnormality are discussed on the light of these and other cases previously reported in the literature.

[The scimitar syndrome in children. Apropos of 3 cases]. / Syndrome du cimeterre de l'enfant. A propos de 3 observations.

The authors describe three new cases in children of abnormal right pulmonary venous drainage into the inferior vena cava, associated with sequestration of the right lower lobe and dextrocardia caused by right pulmonary hypoplasia with a scimitar sign. The term scimitar syndrome used in the literature is to some extent inaccurate as it is neither constant nor pathognomonic. The authors have reviewed 45 paediatric cases reported in the literature and compared the different clinical, diagnostic and therapeutic aspects with their patients. To their knowledge they report the first case in childhood with right lobar sequestration, which received its blood supply from an abnormal artery arising from the right internal mammary artery.