Selection of housekeeping genes for normalization by real-time RT-PCR: analysis of Or-MYB1 gene expression in Orobanche ramosa development.

Real-time PCR has become the method of choice for accurate and in-depth expression studies of candidate genes. To avoid bias, real-time PCR is referred to one or several internal control genes that should not fluctuate among treatments. A need for reference genes in the parasitic plant Orobanche ramosa has emerged, and the studies in this area have not yet been evaluated. In this study, the genes 18S rRNA, Or-act1, Or-tub1, and Or-ubq1 were compared in terms of expression stability using the BestKeeper software program. Among the four common endogenous control genes, Or-act1 and Or-ubq1 were the most stable in O. ramosa samples. In parallel, a study was carried out studying the expression of the transcription factor Or-MYB1 that seemed to be implicated during preinfection stages. The normalization strategy presented here is a prerequisite to accurate real-time PCR expression profiling that, among other things, opens up the possibility of studying messenger RNA levels of low-copy-number-like transcription factors.

Congenital Harlequin syndrome as an isolated phenomenon: A case report and review of the literature.

UNLABELLED: Harlequin syndrome (HS) is a rare autonomic disorder due to a hemifacial cutaneous sympathetic denervation. It is characterized by unilateral diminished sweating and flushing of the face even though after heat or prolonged exercise. It is typically acquired. Congenital cases only represent a 6% of all individuals with HS. All congenital HS cases reported so far showed a concomitant Horner syndrome, probably due to a stellate ganglion involvement. HS represents an uncommon autonomic disorder due to a hemifacial cutaneous sympathetic denervation. It is clinically characterized by a dramatic alteration in facial appearance: ipsilateral denervated pale and dry half from the other intact red and moist half. CONCLUSION: We present, to the best of our knowledge, the first case of a patient with a congenital HS as an isolated phenomenon.

Sources of Resistance to Crenate Broomrape Among Species of Vicia.

Crenate broomrape is a parasitic weed that represents a major constraint for pulse and forage legume production in the Mediterranean and West Asia regions. Control strategies have centered around agronomic practices and the use of herbicides, although success has been marginal. Resistance breeding is hampered by scarcity of proper sources of resistance and of a reliable and practical screening procedure. A germ plasm collection of 208 accessions of vetch belonging to 42 Vicia spp. was screened for resistance to crenate broomrape under field conditions. High levels of resistance were found in several species. Resistance of selected accessions was confirmed by a low induction of broomrape seed germination in pot and in vitro experiments. This was followed by a scarce establishment of broomrape radicles in contact with host roots and a limited development of established tubercles. In addition, a hypersensitive-like necrosis occasionally was observed, but at low frequency.

Luminescent pentafluorophenyl-cycloplatinated complexes: synthesis, characterization, photophysics, cytotoxicity and cellular imaging.

Luminescent mono(pentafluorophenyl) cycloplatinated complexes [Pt(C^N-κC,N)(HC^N-κN)(C6F5)] [HC^N = Hthpy (2-(2-thienyl)pyridine) 2a, Hbt (2-phenylbenzothiazole) 2b, Hpq (2-phenylquinoline) 2c] have been prepared by C­H activation of a HC^N ligand in the corresponding [Pt(HC^N-κN)2(C6F5)2] (1a, 1b, 1c) complexes. Complexes 2 evolve in DMSO solution into solvate complexes and we present here successful routes for the synthesis of [Pt(C^N)(C6F5)(DMSO)] (C^N = thpy 3a, bt 3b). They have been fully characterized (X-ray for 1a, 1c, 2b, 3a and 3b), their electronic absorption and emission properties have been investigated and DFT and TD-DFT calculations for 1a, 1c, 2b and 3a have been carried out. Complexes 3a, 3b and [Pt(ppy)(C6F5)(DMSO)] 4 (Hppy = 2-phenylpyridine) show remarkable stability in a mixed DMSO-cellular medium and their cytotoxicity towards the human lung tumor (A549) and bronchial epithelial non-tumorigenic (NL20) cell lines has been evaluated by MTS assays. Their cellular localization in A549 and NL20 human cells and in mouse embryonic fibroblasts obtained from lungs (LMEFs) has also been investigated by fluorescence microscopy.

Neonatal sepsis and meningitis in a developing Latin American country.

In a retrospective study at Hospital del Niño in Panama City, Panama, 577 neonates with culture-proved sepsis and/or meningitis were identified during an 18-year period (1975 to 1992). Overall there was an incidence of 3.5 cases/1000 live births. Three hundred thirty-three patients (58%) were of low birth weight (< 2500 g) and 260 (45%) were premature. Gram-negative bacilli, particularly species of Klebsiella and Escherichia coli, were responsible for 61% of infections, whereas Gram-positive isolates (especially staphylococci) and Candida strains accounted for 37 and 2%, respectively. The patterns of predominance among bacterial pathogens, however, changed during the period of study. In the later years of this study the frequency of Gram-negative bacteria declined whereas those of staphylococci and Candida increased. Likewise systemic infections caused by Group B Streptococcus organisms appeared recently. The case-fatality rate was 32%. Mortality was greater in infants with early onset sepsis than in those with late infections (44% vs. 22%, P < 0.0001; odds ratio, 2.8; 95% confidence interval, 1.9 to 4.1) and lesser in neonates infected by coagulase-negative staphylococci than in those infected by any other pathogen (12 vs. 39%, P < 0.001; odds ratio, 0.2; 95% confidence interval, 0.1 to 0.4). These findings provide guidelines for the selection of empiric antimicrobial agents in our country and possibly in other Latin American countries and suggest that a continued thorough epidemiologic evaluation is needed to anticipate bacteriologic changes over time.

Sex differences in the effect of MK-801 on normal and spinal cord injured rats.

The induction of functional paraplegia in female rats by contusive spinal cord injury was not prevented by compound MK-801. However, the treatment reduced cavitation around the lesion epicenter to 14 mm3 compared to 17 mm3 in untreated controls t-test, P < 0.28) and conserved more neurons in defined regions outside the lesion epicenter (drug-treated animals vs untreated controls: 299 vs 73 neurons/mm2; t-test, P < 0.009). Thus, although MK-801 was only partially effective in preventing neuronal death secondary to contusion injury it appeared to have a definite neuroprotective effect. In view of the variety of side effects of MK-801 and the controversy on the mechanism of neuroprotection, we examined the action of the drug on non-injured animals. The effects of the drug were strongly sex-dependent. One hour after subcutaneous injection (0.5 mg/kg), female rats were hypothermic (36.8 °C treated vs 38.3 °C control) whereas male rats were hyperthermic (39.6 °C treated vs 38.4 °C control). In females, MK-801 caused cessation of cycling and appearance of numerous polymorphonuclear (PMN) phagocytes in vaginal frotis. Also, beginning 24 h after MK-801 injection, the proportion of PMN increased 400% in female blood, whereas males maintained control values. Arthritis-like joint inflammation was prominent in the toes of female rats, but males were unaffected. After continued treatment with the drug for 15 days, PMN count in female rats decreased and the animals resumed cycling. However, during this period female rats lost 20% of their weight, whereas males gained 26%. One hour after MK-801 injection large increases in blood pressure occurred in both sexes, returning to normal values 2 h later. Hypothermia does not appear to be a factor in the neuroprotective effect of MK-801, but the drug has a number of potentially dangerous side effects, particularly in female rats. Because polymorphonuclear cells are known sources of oxygen free radicals, neuroprotection by MK-801 treatment ought to be much more efficient in males than in females and the drug should be used in combination with a free-radical scavenger.

[Which variables predict the appearance of right bundle-branch block and its course after heart transplantation?]. / Qué variables predicen la aparición de un bloqueo de rama derecha del haz de His y su evolución tras el trasplante cardíaco?

AIM: The purpose of this study was to analyze a series of variables in donors as well as of the preoperative and early and late postoperative of patients developing a right branch block of the bundle of His (RBBBH) in the first week after heart transplantation (HT), and to evaluate factors predicting the disappearance or progression of this conduction disorder. MATERIAL AND METHODS: 58 consecutive patients having undergone an orthotopic HT were studied. 15 of them showed a RBBBH after the HT (age: 43 +/- 13, 12 male, 3 female) and 43 did not (age: 54 +/- 17, 40 male and 3 female). ECGs and echocardiographic studies were performed after 1 week, 1, 3 and 6 months and 1 year. The following factors were analyzed: age and sex of the donor baseline cardiopathy, donor's weight related to recipient's weight, time of ischaemia and cardiopulmonary by-pass, number of rejections per patient/year, previous pulmonary vascular resistance. These parameters were compared among the patients who showed RBBBH and those who did not, and between those whose blocks disappeared in the follow-up and those whose blocks persisted. RESULTS: We found differences is the sex of donors, age of recipients, baseline etiology and time of cardiopulmonary by-pass (with block: 43 +/- 13 years old, dilated cardiomyopathy 73%, 106 +/- 25 minutes, whereas without block: 54 +/- 17 years old, dilated cardiomyopathy 42%, 92 +/- 18). The different parameters between the patients whose block underwent a regression or a progression were sex, lung resistances and right ventricle diameter (progression of the block: men 100%, 3.43% +/- 1.05 UW, progressively growing ventricular diameters. No progression or regression of the block: men 67%, 1.63 +/- 0.74 UW, ventricular diameters with progressive decrease). CONCLUSION: Younger recipients, with a diagnosis of dilated cardiomyopathy, to whom a woman's heart is implanted and who show a longer extracorporal circulation time are those who show a higher incidence of RBBBH: Male patients with high pulmonary resistances undergo a progressive increase in the diameter of the right ventricle and a progressive increase in the RBBBH degree.

[The incidence of major histocompatibility system antigens in dilated and ischemic myocardiopathies]. / Incidencia de los antígenos del sistema mayor de histocompatibilidad en la miocardiopatía dilatada e isquémica.

AIM: The purpose of this study was to analyze the frequency of the different antigens of HLA in patients with diagnosis of very advanced dilated cardiomyopathy and ischemic heart disease by comparing them with a control group of supposedly healthy subjects. MATERIAL AND METHOD: The group of dilated cardiomyopathy consisted of 35 patients (8 women and 27 men) aged between 14 and 60 years. The group of ischemic heart disease included 32 patients (4 women and 28 men) aged between 34 and 64 years. The control group comprised 1337 subjects of the Spanish Mediterranean area, supposedly healthy and recruited from paternity studies. RESULTS: In dilated cardiomyopathy we found a higher incidence in comparison with the control group of the A-2 (62.86% vs 46.22%), B-12 (60.00% vs 32.38%) and DQ-3 (82.86 vs 49.96%) antigens, and a lower incidence of B-51 (0.00% vs 12.49%). In ischemic heart disease we found, when comparing to the control group, a higher incidence of A-11 (31.25% vs 13.08%) and A-29 (34.38% vs 14.58%) antigens and a lower incidence of DQ-2 (15.63% vs 49.88%). CONCLUSIONS: In the Spanish Mediterranean area, the presence of A-2, B-12 and DQ-3 antigens, as well as the absence of B-51 would favour the appearance of advanced dilated cardiomyopathy. The presence of the A-11 and A-29 antigens would predispose to ischemic cardiomyopathy while the presence of DQ-2 would have a protective effect on the appearance of this cardiopathy.

[Doppler echocardiography assessment of cardiac abnormalities in parenteral drug addicts]. / Valoración de las alteraciones cardíacas por ecocardiografía-Doppler de los pacientes adictos a drogas por vía parenteral.

We studied by Echocardiographic-Doppler 114 consecutive intravenous drugs addicts (IVDA); 91 were positive human immunodeficiency virus (HIV+) and 23 negatives. We classified them in five groups; beginning the negative HIV as group 0, and groups I to IV stratified according the Central Disease Control (CDC) classification. We compared the cardiac abnormalities founded between themselves and a control group presumed healthy persons of similar age. The cardiac cavities dimensions showed a statistic significant increased left ventricular end-systolic and diastolic diameters, right ventricular diameter, posterior wall and interventricular septum thickness and aortic root diameter compared with the control group; but all were in the normal range for age. The left ventricular fractional shortening was statistically different from control group related the other groups, and the group IV related other. The existence and severity of pericardial effusions were directly related to the illness stage. We founded moderate pericardial effusions in 25% patients in the 0 to III groups, increasing until 50% in the group IV. The presence of valvular vegetations, nearly 30% in our series, ought to the IVDA. We did not found relationship between the severity of valvular incompetence and the illness stage. We recorded a excellent correlation between the ratio T4/T8 lymphocytes with the progress of illness and the existence and severity of cardiac abnormalities.

[Clinical evaluation of the changes produced by nitrates in patients with chronic heart insufficiency]. / Valoración clínica de los cambios producidos por los nitratos en pacientes con insuficiencia cardíaca crónica.

The aim for this study was to evaluate the hemodynamic changes that appear in patients with myocardiopathy at functional stage II (NYHA) subjected to standard treatment (digital+diuretics) plus nitrates. We used isosorbide-5-mononitrate (IS-5-MN). We studied 20 patients for 100 days in different phases. The placebo-controlled, crossover, double-blind stage random distribution was from day 2 to 5. From day 6 to 20 all the patients took 40 mg/12 hours of IS-5-MN, taking 40 mg/8 hours of active ingredient from day 21 to 100. We performed basal cycloergometrics 3 and 6 hours after administration on days 2, 5, 20, 34 and 100 of the treatment, as well as echocardiograms on these same days. We compared the results obtained versus placebo (p < 0.05). There were no significant differences in heart rate at rest or systolic arterial tension at rest or maximum. Maximum heart rate increased significantly in the tests at days 34 (basal: 143 +/- 9; 3 hours: 139 +/- 8; 6 hours: 140 +/- 7) and 100 (basal: 145 +/- 7; 3 hours: 141 +/- 8; 6 hours: 142 +/- 6) versus the tests carried out with placebo (basal: 136 +/- 8; 3 hours: 135 +/- 7; 6 hours: 136 +/- 7). There were no differences in the tests after acute dose or at treatment day 20.(ABSTRACT TRUNCATED AT 250 WORDS)

[Non specific leukodystrophy. A new case of vacuolizing leukoencephalopathy with megalencephaly]. / Leucodistrofias indeterminadas. Un nuevo caso de leucoencefalopatía vacuolizante con megalencefalia.

INTRODUCTION: Amongst the conditions affecting the white matter, the disorders of myelinization, including the leukodystrophies, are important in the field of paediatric neurology. Although classically they have been classified according to whether the metabolic defect was known or not, at the present time great advances in neuroimaging have clarified many genetic disorders involving the white matter and new classifications have been devised. The group of unknown aetiology includes the so called non specific leukodystrophies, characterized by their onset in infancy with a usually more moderate clinical course, and neuro imaging (computerized tomography CT magnetic resonance MR ) with alteration of the signal from the white matter which is symmetrical, bilateral and diffuse. Study and investigation of the patterns of MR has permitted isolation of two new clinical conditions of the non specific leukodystrophies group: leukodystrophy with megalencephaly and temporal cysts (Van der Knaap, 1995) for which currently the term vacuolizing leukoencephalopathy with megalencephaly is preferred and the CASH syndrome (childhood ataxia with central hypomyelinization or vanishing white matter disease) (Van der Knaap, 1997). DEVELOPMENT: We present a review of nine cases of non specific leukodystrophies with an average course of 13 years. They were studied using the protocol of the European working party on demyelinating diseases. One of these fulfilled clinical and radiological criteria for the diagnosis of vacuolizing leukoencephalopathy with megalencephayl: onset in early childhood, macrocephaly, normal metabolic studies, moderate progression and alteration of the white matter signal which was bilateral, symmetrical and diffuse with the presence of oedema and temporal subcortical cysts. We discuss the most relevant articles currently published on this condition.

[The phenomenon of genomic "imprinting" and its implications in clinical neuropediatrics]. / El fenómeno del 'imprinting' genómico y sus implicaciones en clínica neuropediátrica.

OBJECTIVE: We comment on the most important advances related to the phenomenon of genomic 'imprinting' in clinical paediatric neurology. DEVELOPMENT: Initially, we review the biological findings related to this subject and establish various concepts. Later, we attempt to clarify the different mechanisms of expression of the phenomenon 'imprinting' and its application in clinical practice. We give a detailed review of the various neurological disorders in which this genetic phenomenon has been involved to date. Finally, we attempt to determine when this genetic alteration should be suspected and which molecular biology techniques should be used to confirm the diagnosis. CONCLUSIONS: 1. Clinical diagnosis suspecting that the presence of genomic 'imprinting' may be the mechanism causing a particular pathology should be based on a family tree showing that both sexes and all generations are affected and that the severity of the same disease varies among different members of the same family; 2. Study strategy includes studying the methylation pattern of the DNA. If there are changes in this, PCR should be done to show the exact pattern of the alteration.

[Cerebral hemorrhage in full term newborns]. / Hemorragias cerebrales en el recién nacido a término.

INTRODUCTION: Cerebral hemorrhages in full-term newborn babies are an important factor in neonatal morbidity and mortality and very frequent in premature babies. In full-term newborn the frequency is reduced to 1-2% and the aetiopathogenesis is basically related to birth trauma. OBJECTIVE: To identify the clinical forms of cerebral hemorrhages in full-term newborn before taking prophyllactic and/or therapeutic measures if possible. DEVELOPMENT: Based on the integrated physio-pathological model of Wigglesworth and Pape, two anatomo-pathological patterns were established according to gestational age. Topographic classification was done in full-term newborn according to site (subarachoid, subdural, intraventricular, cerebellar and intraparenchymatous). We studied the pathogenesis, clinical features and diagnosis of each of these sites and emphasize the importance of neuroimaging. CONCLUSIONS: The diagnostic approach proposed permits an aetiopathogenic and therapeutic view which currently permits improved prognosis and even cure in many cases.

[Epileptic signs in alterations of neuronal migration]. / Manifestaciones epilépticas en las alteraciones de la migración neuronal.

INTRODUCTION: A wide range of conditions are due to alterations in neurone migration (ANM). Mental retardation, motor disorders and epilepsy are seen in all these disorders. Anomalies included with the ANM are those produced at the time of neuronal migration in the phase prior to neurone proliferation and during the time following cortical organization. All these have a common characteristic, namely an anomalous cerebral cortex (cerebral dysplasias). DEVELOPMENT: There is a high incidence of epilepsy in ANM (60%), appearing early (10% neonatal, 42% before 1 year old). The frequency of crises increases with age. Extended and diffused generalized forms presents as epileptic encephalopathies (Ohtahara, West, Lennox-Gastaut), whilst focal forms are seen as simple motor, partial crises, complex and secondarily generalized crises with a tendency to status epilepticus and also to continuous partial epilepsy. In diffuse, extensive forms, the EEG is characterized by large amplitude theta-delta rhythm activity (specific) with the presence of rapid activity (15-25 Hz) and of large amplitude (150-300 microV) which may also be found in other processes. In localized forms the recordings vary: localized discharges with/without crises, multifocal discharges in more than two lobes with a defined critical area, positive discharges, ipsilateral spike-and-wave complexes associated with focal discharges or with normal recordings. We review different types of ANM: in the phase of proliferation (hemimegalencephalia), the agyria-paquigyria complex; in phase of migration: type I lissencephalias (Miller-Dieker), layered heterotopias (double cortex), type II (cobblestone) lissencephalias and neuronal heterotopias, and in the phase of organization of the cortex: polymicrogyria and the esquissencephalias I and II. CONCLUSIONS: The functional prognosis in ANM depends on the control of the crises rather than on the extent of the lesion. Surgical treatment leads to 42% good or excellent results.