Comparison of two different protocols of neonatal screening for cystic fibrosis.

The results of two different protocols of neonatal cystic fibrosis (CF) screening in the Lazio region of Italy are reported. The first study, conducted from 1992 to 2000 on about 200,000 newborns, consisted of an immunoreactive trypsin (IRT) protocol without mutation analysis of the cystic fibrosis transmembrane conductance regulator (CFTR) gene, referred to as the IRT/IRT protocol. Approximately 5% of the newborns with a positive first IRT test were also positive at the second test; approximately 57% of the newborns with a high IRT level at the second test were subsequently found to be affected by CF. In September 1998, a second protocol that included mutation analysis (IRT/DNA/IRT protocol) was started. Comparison of the two different screening protocols in terms of sensitivity in detecting CF patients demonstrated that the IRT/DNA/IRT protocol is more effective because it is able to detect a higher number of CF patients than the IRT/IRT protocol. In the same period, in addition to the overall diagnosis performed on a screening basis, 64 other subjects were identified as being affected by CF on the basis of symptomatic findings. The overall incidence of CF (screening + symptoms) was 1 : 2982, while that for carriers was 1 : 27. The sensitivity of the screening program increased over the period from 1992 to 2000, with the enhanced sensitivity in the past 2 years being due to the introduction of the IRT/DNA/IRT protocol.

Terbinafine in the treatment of non-immunocompromised compassionate cases of bronchopulmonary aspergillosis.

Conventional treatments of broncho-pulmonary aspergillosis are often ineffective and result in associated side-effects. Terbinafine (a new allylamine derivative), although as active against Aspergillus in vitro as amphotericin B and itraconazole, is less effective in rodent models because of a rapid hepatic first-pass effect. As terbinafine is metabolized differently in humans, the aim of this work was to evaluate this drug, for the first time, in the treatment of seven immunocompetent patients with lower respiratory tract mycotic infections unresponsive to the usual antimycotic drugs. Diagnosis was based on identification of fungal isolates, worsening of respiratory function tests, chest radiographs and computerized tomographic (CT) scan changes, positive skin test, aspergillin precipitins and clinical history. Terbinafine was administered at doses ranging from 5 to 15 mg kg-1 day-1 depending on the clinical severity of the disease, and was given for 90-270 days depending on clinical progress and compliance. In three patients A. fumigatus was suppressed with resolution of signs and symptoms; four patients showed transitory A. fumigatus suppression with marked clinical and radiological improvement. During relapses no resistance to terbinafine was observed. No significant side-effects were detected. Terbinafine appeared to be as effective as amphotericin B and itraconazole in the treatment of bronchopulmonary aspergillosis in nonimmunocompromised patients. These preliminary results suggest that controlled studies are warranted.

Deletion analysis of the simple tandem repeat loci physically linked to the spinal muscular atrophy locus.

Multicopy dinucleotide repeats have been characterized in the spinal muscular atrophy (SMA) region on chromosome 5q13, which reveal deletions in some SMA patients. 119 Italian and Spanish SMA families have been analysed using the C272 and C212 markers. Seventy percent of these families were informative. We found 9.4% de novo deletions in SMA I and 1.5% in SMA II families. A single inherited deletion segregating in a Spanish pedigree was detected in three affected brothers. A SMA II patient showed deletion only of C272. The data presented in this study are relevant to the molecular diagnosis of SMA families in Italy and Spain and provide additional insights toward the understanding of the molecular pathology of SMA.

Identification of multiple transcribed sequences from the spinal muscular atrophy region of human chromosome 5.

We report the isolation and characterization of novel expressed sequences from the spinal muscular atrophy (SMA) region on human chromosome 5q13. Based on the sequence homology studies these cDNAs were grouped in four classes, one of which shows extensive homologies with the beta-glucuronidase (BG) gene, differing in exon arrangement. The other cDNAs do not show any strong homology with known DNA sequences.

First-trimester prenatal diagnosis of spinal muscular atrophy using microsatellite markers.

Twenty-five pregnancies at risk for spinal muscular atrophy I (SMA I) have been monitored by first-trimester prenatal diagnosis. Microsatellite markers were used in all cases to amplify polymorphic regions at the D5S125, D5S435, D5S39, D5S127, and D5S112 loci. All families, including 12 SMA I pedigrees with a decreased index child, were fully informative for DNA analysis. Three fetuses were predicted to be affected and 22 fetuses were predicted to be unaffected. Twenty-two newborns were unaffected by clinical examination at birth. These results support the accuracy of SMA I prenatal diagnosis based on linkage analysis.

Human elongation factor EF-1 beta: cloning and characterization of the EF1 beta 5a gene and assignment of EF-1 beta isoforms to chromosomes 2,5,15 and X.

We report here the isolation and characterization of a novel human elongation factor-1 beta (EF-1 beta) gene by cDNA selection from YAC mapping on chromosome 5q12-q14. This gene is specifically transcribed in fetal brain and in skeletal muscle and is characterized by a complete sequence homology with previously described EF-1 beta cDNAs. We also assigned the loci for three other EF-1 beta isoforms, to human chromosomes 2, 15 and X. The multiple chromosomal assignments of EF-1 beta loci demonstrates the genetic heterogeneity of human EF-1 beta peptides.

Isolation and cloning by a polymerase chain reaction of a genomic DNA fragment of the human slow skeletal troponin (TNNT1) gene.

The genomic 3' structure of the gene coding for the human slow skeletal troponin T (TNNT1) gene, is reported. An intron of 912 nucleotides containing an Alu-element has been identified and characterized. The complexity of the sequenced region suggests an alternative exon use. The present results may be valuable for further studies on the gene structure of TNNT1 and the related troponin gene family.

A tool for the molecular analysis of an early lethal disease: slide-PCR in spinal muscular atrophy patients.

DNA was recovered from sections of muscle biopsies of 20 spinal muscular atrophy (SMA) patients fixed on microscopic slides and stored from one to 20 years at room temperature. Microsatellite DNA markers tightly linked to the SMA locus were amplified using the polymerase chain reaction (PCR) to obtain specific amplified products. The procedure was successful in all cases, and allowed prenatal diagnosis in one at-risk pregnancy. In our hands this procedure is quick, sensitive and reproducible.

Polymerase chain reaction in the detection of mRNA transcripts from the slow skeletal troponin T (TNNT1) gene in myotonic dystrophy and normal muscle.

Recent studies have shown that the gene encoding for the slow skeletal troponin isoform T (TNNT1) is located on the proximal long arm of human chromosome 19 in the myotonic dystrophy (DM) region. In order to test TNNT1 as a candidate gene for DM, we have isolated TNNT1 cDNA from skeletal muscle from two healthy individuals and from two patients with DM. Sequencing of the TNNT1 cDNA from the DM and normal muscle revealed two sequence variants but no transcriptionally significant mutations. This work rules out a defect in the coding segment of TNNT1 as a cause of DM and provides a polymerase chain reaction protocol for studying troponin T gene expression.

Legionnaire's pneumonia complicating a thermal burn.

The report describes a patient with 45 per cent BSA burns who developed Legionnaire's disease 3 days after the acute injury. The diagnosis of this life-threatening complication was late because most of its signs and symptoms can be encountered in the burned patient. This delay could have been fatal to the patient and required the evacuation of the burn centre for disinfection.

Trace of metal exposure in hard metal lung disease.

A male subject exposed for many years to metal dusts from grinding and cutting hard metals was investigated four years after the exposure ceased. While chest x-ray films and histologic examinations showed diffuse interstitial fibrosis and severe perivascular and peribronchiolar fibrosis, radiochemical neutron activation analysis (NAA) showed high W, Ta and Co concentrations in the pulmonary specimen compared to the corresponding determinations in 17 control subjects. Tantalum was also found in high concentrations in bronchoalveolar lavage fluid (BAL) and in the blood. In addition other elements such as Ni, Cr, Th and Cd were found in the lung biopsy specimen of the worker in amounts significantly higher than in the control subjects. The results suggest that hard metal origin of pulmonary fibrosis can be detected many years after removal from exposure.

Effects of captopril on hemodynamics and blood gases in chronic obstructive lung disease with pulmonary hypertension.

The effects of Captopril, an angiotensin-converting enzyme inhibitor, on pulmonary hemodynamics and blood gases were studied in 9 patients with chronic obstructive lung disease (COLD) and pulmonary hypertension (PA-P greater than 20 mm Hg). Hemodynamic data were recorded prior to Captopril administration (50 mg per os) and for the next 60 min. Following Captopril administration, significant reductions in mean pulmonary artery pressure (PA-P) (p less than 0.05), in mean pulmonary wedge pressure (PW-P) (p less than 0.05), and in total pulmonary resistance (TPR) were noted; significant reductions in mean brachial artery pressure (BA-P) and systemic vascular resistance (SVR) were also recorded, while cardiac output, heart rate and blood gas tensions showed no significant changes. Furthermore, the higher the hypoxemia, the greater was the reduction in BA-P (p less than 0.05). We therefore feel that Captopril, when administered to COLD patients with pulmonary hypertension, may protect the pulmonary circulation from hypoxic pulmonary vasoconstriction.

Influence of ACE inhibition on pulmonary haemodynamics and function in patients in whom beta-blockers are contraindicated.

The effects of captopril have been studied in 19 patients suffering from chronic obstructive pulmonary disease (COPD). In 9 of these patients with pulmonary hypertension (mean pulmonary artery pressure greater than 20 mm Hg), we studied the effects of the drug on pulmonary haemodynamics, blood gases and systemic circulation. Haemodynamic data were recorded before oral captopril 50 mg and for the next 60 minutes. Following captopril administration, significant reductions in mean pulmonary artery pressure (PAP) (P less than 0.05), in mean pulmonary wedge pressure (PWP) (P less than 0.05) and in total pulmonary resistance (TPR) were noted; significant reductions in mean brachial artery pressure (BAP) and systemic vascular resistance (SVR) were also recorded, while cardiac output, heart rate and blood gas tensions showed no significant changes. Furthermore, the higher the hypoxaemia was, the greater the reduction in BAP (P less than 0.05). In the other 10 COPD patients with moderate essential hypertension, captopril was given as monotherapy (75-10 mg/day) for 60 days. We found no significant modification of the various respiratory function tests except for an increase in the vital capacity (P less than 0.05). Systolic blood pressure and diastolic blood pressure were reduced both in the supine and in the standing position and there were no side effects, in particular no bronchospasm even in the patients responsive to bronchodilator drugs. Our data suggest that captopril is an effective and safe drug when administered to COPD patients with essential hypertension. Moreover, in these patients, it may protect the pulmonary circulation from hypoxic pulmonary vasoconstriction.

Can pulmonary hypertension be predicted by non-invasive approach? Echocardiographic and haemodynamic study.

41 patients suffering from Chronic Obstructive Lung Disease (COLD) and 44 with Sarcoidosis were studied. Said patients underwent respiratory function tests, echocardiography (M.mode) to assess the right ventricular index ( RVI = diameter of the right ventricular cavity corrected by body surface) and the thickness of the right ventricular anterior wall ( RVAWT ); patients also underwent right heart haemodynamics (Swan-Ganz catheter). These data were further statistically studied by means of multiple regression in order to assess the eventuality of a non-invasive prediction of pulmonary artery mean pressure (PAP): variables taken into consideration were: age, body surface (BS), RVI , RVAWT , arterial oxygen pressure (PaO2), arterial carbon dioxide pressure (PaCO2) and PAP dependence according to each case group and the interaction of each group itself on the variables. RVI appeared to be the most reliable, in fact, when PaCO2 is also available, the standard error of estimation (SEE) was only 3.84 mmHg and the coefficient of determination was equal to 85.5% with a notable improvement when compared to results seen in previous studies. This behaviour was observed both in patients with early sarcoidosis and in COLD patients with mild pulmonary hypertension. This might be due to the fact that we took the right ventricle into consideration which inevitably feels the increase in pulmonary hypertension.

Pharmacological control of platelet-vessel wall interactions in patients with chronic obstructive airway disease.

In the last few years great interest has been devoted to the role of platelet activation in the haemodynamic and thrombotic complications which frequently occur in patients with chronic obstructive airway disease. Several recent studies have shown, in fact, a number of alterations of platelet function in this pathological condition, such as an increased betathromboglobulin level and higher levels of malondialdehyde after collagen and thrombin platelet stimulation compared to age and sex matched control subjects. Our study indicates that the platelet regeneration time is shortened in patients with chronic obstructive airway disease in respect to that of controls (p less than 0.001). The administration of dipyridamole (425 mg/die) resulted in a significant prolongation of the platelet regeneration time in these patients indicating that the drug was effective in reducing the increased platelet turnover.

Assessment of sarcoidosis activity by 67gallium lung scan. A study with follow-up.

71 consecutive patients with histologically confirmed sarcoidosis, in various clinical stages of activity, were submitted to 67Ga lung scan, and 23 of them were studied with two or more scans at intervals of 4-6 months. In patients on steroid therapy, the drug was suspended 7 days before scan to avoid the steroids interfering with the gallium (Ga) uptake mechanism. In order to assess the usefulness of 67Ga in the evaluation of sarcoid activity, six other parameters of activity were considered, ranging from angiotensin-converting enzyme levels to progressive symptoms, from deteriorating X-ray or pulmonary function tests to clinical or laboratory evidence of prominent extra thoracic involvement. Our work suggests that Ga scan is more sensitive than chest X-ray in determining the degree and variation of pulmonary sarcoidosis activity, in evaluating the response to therapy, and in foreseeing the relapses. In some cases it gives information not detectable with other noninvasive criteria. The detection of patients with active disease, after discontinuation of steroids 7 days before scan, raises doubts about the opportunities of scanning patients on steroids, and suggests that further studies on this point are needed.

Echocardiographic and hemodynamic assessment of right heart impairment in chronic obstructive lung disease.

42 patients with chronic obstructive lung disease underwent right heart hemodynamics (Swan-Ganz catheter) and M mode echocardiography. Echocardiographic study showed that right ventricular index (RVI) and right ventricular anterior wall thickness (RVAWT) were increased in most patients, while septal thickness and motion were nearly always normal. The finding of impaired RVI and normal RVAWT in several patients, while no one showed normal RVI and increased RVAWT, suggests that arterial pulmonary hypertension results in initial dilatation and only later in hypertrophy of the right ventricle. Echocardiographic parameters were well correlated with hemodynamic and pulmonary function parameters. The good correlations between echocardiographic and hemodynamic data suggest that echocardiography may be a useful technique in the noninvasive assessment of the effects of pulmonary hypertension on the right heart in COLD.