The anti-proliferative effects of adiponectin on human lung adenocarcinoma A549 cells and oxidative stress involvement.

Adiponectin (Acrp30) plays an important role in energy metabolism and inflammation. Recently, in vivo serum Acrp30 levels have been reported to be correlated to risk of developing several types of cancers such as lung cancer, and in vitro studies have demonstrated a role for Acrp30 in the control of cell proliferation and survival. However, the molecular effects of Acrp30 on lung cancer have not yet been clearly defined. In the present study, we investigated the effects of different concentrations of Acrp30 on the A549 human alveolar epithelial cell line, an in vitro model of lung adenocarcinoma. A549 cells were exposed to various concentrations of Acrp30 and successively, proliferation, apoptosis and oxidative stress were evaluated by MTT test, caspase activity assay, flow-cytometry and western blotting analysis. Our results demonstrated that Acrp30 causes, in a time- and dose-dependent manner, a reduction of cell viability and duplication together with an increase in cell apoptosis rate. In addition, we found that Acrp30 induces an increase of lipid peroxidation evaluated by TBARS assay and a concomitant reduction of nitric oxide release, both markers of cellular oxidative stress. Taken together, our data on A549 cells provides new insight into potential involvement of Acrp30 on physio-pathologic mechanisms of lung diseases through interference with proliferation, apoptosis and oxidative status.

Nitric oxide in upper airways inflammatory diseases.

In the human respiratory tract, the main production sites of exhaled nitric oxide (NO) are the nose and paranasal sinuses. In the upper airways, NO has been suggested to be involved at different levels with regulatory, protective, defensive or deleterious effects. Therefore, we review some aspects of the origin, metabolism, and functions of NO in the upper airways, together with the role of NO in some upper airways inflammatory diseases. Furthermore, we discuss the recent improvements in nasal NO measurements, which may be useful to better characterize the involvement of the NO produced by nose and paranasal sinuses in upper airways inflammatory diseases such as allergic rhinitis, nasal polyposis, sinusitis, primary ciliary dyskinesia, and cystic fibrosis.

Right ventricular performance in severe obesity. Effect of weight loss.

BACKGROUND: The effects of severe obesity on right ventricular function in the absence of associated cardiopulmonary disease are not well known. Right myocardial performance index (R-MPI) is an echocardiographic index to non-invasively assess the right ventricular function. The aim of our study was to assess R-MPI in individuals with severe but uncomplicated obesity before and after a significant weight loss induced by bariatric surgery. PATIENTS AND METHODS: Fifteen obese females (OB) without cardiovascular and pulmonary diseases were examined. In all subjects, R-MPI was calculated by Doppler echocardiography as the sum of isovolumetric contraction time and isovolumetric relaxation time divided by ejection time. Furthermore, pulmonary function test (PFT) and 6-min walking test (6mWT) were performed. Ten healthy subjects with normal weight (HS) were also evaluated as controls. R-MPI, PFT and 6mWT were also re-evaluated one year later in 12 obese subjects treated with gastric banding after a consistent weight loss (> 20%). RESULTS: A prolongation of R-MPI was found in OB before bariatric surgery in comparison to the HS (0.47 +/- 0.04 and 0.29 +/- 0.05, respectively; P < 0.001). R-MPI significantly improved in OB 12 months after surgery (0.32 +/- 0.03) and was no longer different from HS. R-MPI positively correlated to body mass index (BMI). A significant association was found between the reduction of BMI after bariatric surgery and the distance walked during the 6mWT. CONCLUSIONS: These results show a right ventricular dysfunction in severe uncomplicated obesity, associated with an impaired functional capacity which recovers after consistent weight loss.

Nasal nitric oxide measurements before and after repeated humming maneuvers.

BACKGROUND: It has been recently shown that humming greatly increases nasal nitric oxide (NO). This is most likely owing to a rapid washout of sinus NO caused by the oscillating sound waves. During repeated humming manoeuvres nasal NO gradually decreases, likely because NO accumulated in the sinuses is washed out. AIM: We studied whether humming before measurements would affect nasally exhaled NO. MATERIALS AND METHODS: NO output was measured by the chemiluminescence technique in orally and nasally exhaled air in 38 subjects: 18 healthy subjects (HS), 15 subjects with allergic rhinitis (AR) and five subjects with allergic nasal polyposis (AP). Each subject performed a NO measurement during quiet nasal exhalation either preceded by a period of silence/free speaking or immediately after five consecutive humming manoeuvres (posthumming). RESULTS: Mean nasal NO output (95% CI) after a period of silence/free speaking was 231 nL min-1 (178-284) in HS, 434 nL min-1 (347-522) in AR (P < 0.001) and 262 nL min-1 (163-361) in AP. Post-humming nasal NO output was 16% (5 to 50%) lower in HS and 14% (1 to 49%) lower in AR, while it remained unchanged in AP subjects. Intra-subject coefficient of variation of quiet nasal exhalation was 12% in HS, 13% in AR and 5% in AP. Post humming intraindividual coefficient of variation significantly decreased in both HS and AR, but it did not change in AP. CONCLUSIONS: Nasal NO levels measured immediately after repeated humming manoeuvres are consistently lower and more reproducible than nasal NO levels measured after a period of silence or free speaking. Repeated humming effectively empties the sinuses, thereby probably minimizing the normal contribution from the sinuses to nasal NO. This may be useful to better estimate NO output from the nasal cavity mucosa in health and disease.

Exhaled nitric oxide after inhalation of isotonic and hypotonic solutions in healthy subjects.

Airway nitric oxide (NO) homoeostasis is influenced by chemical and mechanical stimuli in humans; airway epithelium, which is an important site of NO production, is sensitive to osmotic challenge. The effect of inhaled hypotonic solutions on exhaled NO (eNO) is not known. In this study we evaluated the effect of ultrasonically nebulized distilled water (UNDW), a hypotonic indirect stimulus, on eNO levels. A total of 10 non-smoking healthy subjects were enrolled in the study. eNO was detected by chemiluminescence, and specific airway conductance (sGaw) was measured by plethysmography. Bronchial challenges with UNDW and with an isotonic solution were performed according to a double-blind experimental design. Baseline levels of eNO were 28.1+/-14.7 p.p.b. UNDW did not cause any significant change in sGaw (from 0.190+/-0.029 to 0.181+/-0.036 cm H(2)O x s(-1)). With respect to baseline values, the eNO concentration decreased significantly after inhalation of 8 or 16 ml of UNDW (from 26.0+/-13.1 to 17.2+/-8.5 and 16.6+/-7.7 p.p.b. respectively; P<0.001, n=10). After bronchial challenge with UNDW, eNO was significantly reduced in comparison with after inhalation of the isotonic solution. In five subjects, pretreatment with N(G)-nitro-L-arginine methyl ester (L-NAME), an inhibitor NO synthesis, decreased NO levels from 21.7+/-8.5 to 10.0+/-3.3 p.p.b. Subsequent inhalation of 16 ml of UNDW did not cause any further decrease in NO levels (10.1+/-3.7 p.p.b.; not significant compared with L-NAME). We conclude that inhalation of aqueous solutions decreases eNO levels in healthy subjects, and that this effect is not associated with any significant change in airway calibre. The UNDW-induced decrease in eNO is not enhanced by pretreatment with the NO synthase inhibitor L-NAME, suggesting that inhaled solutions may interfere with the airway NO pathway in humans.

Systematic use of dystrophin testing in muscle biopsies: results in 201 cases.

We studied dystrophin with both immunohistochemistry and immunoblotting in 201 muscle biopsies stored in liquid nitrogen during the period 1985-92. The systematic use of dystrophin testing combined with DNA analysis and with 3-10 years follow-up of the patients yielded a significant modification of the diagnoses made previously and identified dystrophinopathies with unusual expression and course. Seventeen out of 152 (11.18%) diagnoses in males and 8 out of 49 (16.32%) in females were modified by dystrophin testing. Most diagnostic errors (9 out of 27 diagnoses) were in the group Becker muscular dystrophy-limb girdle muscular dystrophy, confirming the clinical overlap of the two diseases. Unusual expressions of dystrophinopathy included muscular dystrophy with early elbow contractures (two patients), recurrent myoglobinuria (one patient), dilating cardiomyopathy (two patients), myoglobinuria and associated dilating cardiomyopathy (one patient), very late-onset benign myopathy (two patients and one manifesting carrier) and congenital myopathy (one manifesting carrier). In the group 'idiopathic hyper-CKaemia', we did not find any dystrophinopathy in 34 males, whereas five out of nine females were found to be carriers. Immunohistochemical analysis of dystrophin using the monoclonal antibody against the C-terminus detected 99% of protein defects and was found to be the most cost-effective way of revealing dystrophinopathies. The combined use of immunohistochemical analysis with the antibody against the C-terminus and immunoblotting with the antibody against the core of the protein appears to be a highly reliable diagnostic approach (100% detection rate).

Unusual expression and very mild course of Xp21 muscular dystrophy (Becker type) in a 60-year-old man with 26 percent deletion of the dystrophin gene.

A 54-year-old farmer with a negative family history had had mild proximal weakness for the previous 4 years. Clinical examination showed marked scoliosis, barrel-shaped chest, diffuse hypotrophy, and mild proximal weakness. Creatine kinase was 938 U/l; electrocardiography and echocardiography were normal. EMG disclosed myopathic changes. Muscle biopsy showed slight, nonspecific alterations. Dystrophin was present and normally distributed with antibodies against the C-terminal and N-terminal, whereas it was not recognized by the antibody against the rod domain. Western blotting detected an abnormal molecular weight protein of 320 kd (normal, 427 kd). Southern blot analysis revealed a deletion from exon 21 to exon 44, corresponding to 26% of the coding region of dystrophin. Six years' follow-up did not disclose progression of the muscle disease.

Metachromatic dye-Ca++ATPase method in pathological muscle: a study of 382 muscle biopsies.

The metachromatic dye-Ca++ATPase method, that in normal muscle distinguishes fiber types on the basis of their metachromatic or orthochromatic staining, was applied to 382 pathological muscle biopsies. Results were compared in serial sections with those obtained with the conventional ammonium sulphide method. In pathological muscle the metachromatic dye-Ca++ATPase method confirms the advantages already showed in normal muscle: fast and easy performance, neat fiber typing, simultaneous staining of nuclei. Moreover in pathological muscle the metachromatic dye-Ca++ATPase method showed some muscle changes which are missed by the conventional ammonium sulphide one, namely central nuclei, macrophagic invasion and some structural abnormalities. This allows immediate correlation between those alterations and fiber typing.

Congenital muscular dystrophy associated with familial junctional epidermolysis bullosa letalis.

A 20-year-old patient was born with epidermolysis bullosa and a severe, slowly progressive muscle disease. Skin biopsy demonstrated junctional epidermolysis bullosa. Muscle biopsy demonstrated degenerative changes with increase in connective tissue, fibre size variability, rods and cytoplasmic bodies, central nuclei. In muscle biopsy dystrophin, chondroitin unsulphate, chondroitin 4-sulphate, chondroitin 6-sulphate, heparan sulphate, collagen III, collagen IV and VI, laminin, and fibronectin were normally distributed. This is the first report of the association of a form of congenital muscular dystrophy with junctional epidermolysis bullosa and, together with the previous reports of muscle involvement in epidermolysis bullosa simplex and dystrophica, it suggests the existence of a syndrome characterized by the contemporaneous presence of skin and muscle involvement.

Clinical spectrum of McArdle disease: three cases with unusual expression.

Three cases of myophosphorylase deficiency with unusual clinical expression are presented. The 1st had clinical characteristics suggesting a mild congenital myopathy, and the patient never experienced cramps or myalgias. The 2nd had a slowly progressive myopathy without cramps or myoglobinuria which was detected by chance. The 3rd presented with myoglobinuria and acute renal failure, unrelated to a triggering effort, and with permanent weakness and wasting. In all cases, muscle biopsy demonstrated a vacuolar myopathy with free glycogen increase and absence of myophosphorylase activity, confirmed by biochemical assays. The cases confirm the wide clinical spectrum of McArdle disease.

Familial autosomal recessive rigid spine syndrome with neurogenic facio-scapulo-peroneal muscle atrophy.

Two sisters and a first cousin presented with rigid spine and facio-scapulo-peroneal muscle atrophy. The patients belonged to a family with two first-cousin marriages. Electromyography, muscle and nerve biopsy showed neurogenic muscle atrophy without peripheral nerve involvement. Follow up did not show progression of the disease. This is the first observation of an association of neurogenic facio-scapulo-peroneal and rigid spine syndrome. The double first-cousin marriage suggests autosomal recessive inheritance.

Cytochrome c oxidase and coenzyme Q in neuromuscular diseases: a histochemical study.

Cytochrome c oxidase (CCO) has been histochemically studied in 250 muscle biopsies from patients with different neuromuscular diseases. The results were compared with those obtained on serial sections stained with Gomori's trichrome and with the methods for NADH tetrazolium reductase, succinate dehydrogenase and lactate dehydrogenase. In 58 selected cases serial sections were also stained with a method demonstrating coenzyme Q (CoQ) activity. Demonstration of structural alterations was as good with CCO as with the methods for other oxidative enzymes: particularly evident were alterations of the distribution of mitochondria, such as core areas in central core and multiminicore diseases. Unstained fibers were observed in mitochondrial myopathies, in Becker, Emery-Dreifuss, limb-girdle, facio-scapulo-humeral muscular dystrophies, muscle infarction, polymyositis, motor neuron diseases and neuropathies. The histochemical method for CoQ showed only low specificity, since partial staining was also present in areas devoid of mitochondria, such as cores. CoQ deficiency was not observed in any of the 19 mitochondrial myopathies examined.