Component-specific toxic concerns of the inhalable fraction of urban road dust.

Continuous global urbanisation causes an ever-growing ecological footprint of pollution. Road dust (RD), one of these pollutants, poses a health concern due to carcinogenic and toxic components potentially present in the micron-sized fractions. The literature reports on the concentrations of trace, toxic metals and metalloids present in RD (Hooker and Nathanail in Chem Geol 226:340-351, 2006), but the literature on its molecular composition is limited. Recent reports on the bioaccessibility of platinum group metals are also reported (Colombo et al. in Chem Geol 226:340-351, 2008). In vitro and animal toxicological studies confirmed that the chemical composition of inhaled particles plays a major role in its toxic, genotoxic and carcinogenic mechanisms, but the component-specific toxic effects are still not understood. Particle-bound airborne transition metals can also lead to the production of reactive oxygen species in lung tissue; a special concern amongst particularly susceptible cohorts (children and elderly). The characterisation of the molecular composition of the fine fraction is evidently of importance for public health. During a pilot study, partially characterised size-fractioned RD samples (Barrett et al. in Eviron Sci Technol 44:2940-2946, 2010) were analysed for their elemental concentration using X-ray fluorescence spectrometry and inductively coupled plasma mass spectrometry. In addition, separately dispersed particles (200 particles per size fraction) were analysed individually by means of computer-controlled electron probe X-ray micro-analysis (CC-EPXMA) and their molecular structure probed by studying elemental associations. These were correlated with micro-Raman spectroscopy (MRS) results. It was found that the fine fraction (<38 µm) had the highest Pb (238 ppm) and Cr (171 ppm) concentrations. The CC-EPXMA data showed >50 % association of Cr-rich particles with Pb, and the MRS data showed that the Cr was mostly present as lead chromate and therefore in the Cr(VI) oxidation state. Concentrations of both Pb and Cr decreased substantially (279 (<38 µm)-13 ppm (<1 mm); 171 (<38 µm)-91 ppm (< 1 mm), respectively) in the larger fractions. Apart from rather alarmingly high concentrations of oxidative stressors (Cu, Fe, Mn), the carcinogenic and toxic potential of the inhalable fraction is evident. Preliminary bioaccessibility data indicated that both Cr and Pb are readily mobilised in artificial lysosomal liquid and up to 19 % of Cr and 47 % of Pb were released.

Acetazolamide prevents vacuolar myopathy in skeletal muscle of K(+) -depleted rats.

BACKGROUND AND PURPOSE: Acetazolamide and dichlorphenamide are carbonic anhydrase (CA) inhibitors effective in the clinical condition of hypokalemic periodic paralysis (hypoPP). Whether these drugs prevent vacuolar myopathy, which is a pathogenic factor in hypoPP, is unknown. The effects of these drugs on the efflux of lactate from skeletal muscle were also investigated. EXPERIMENTAL APPROACH: For 10 days, K(+)-depleted rats, a model of hypoPP, were administered 5.6 mg kg(-1) day(-1) of acetazolamide, dichlorphenamide or bendroflumethiazide (the last is not an inhibitor of CA). Histological analysis of vacuolar myopathy and in vitro lactate efflux measurements were performed in skeletal muscles from treated and untreated K(+)-depleted rats, and also from normokalemic rats. KEY RESULTS: About three times as many vacuoles were found in the type II fibres of tibialis anterioris muscle sections from K(+)-depleted rats as were found in the same muscle from normokalemic rats. In ex vivo experiments, a higher efflux of lactate on in vitro incubation was found in muscles of K(+)-depleted rats compared with that found in muscles from normokalemic rats. After treatment of K(+)-depleted rats with acetazolamide, the numbers of vacuoles in tibialis anterioris muscle decreased to near normal values. Incubation with acetazolamide in vitro inhibited efflux of lactate from muscles of K(+)-depleted rats. In contrast, bendroflumethiazide and dichlorphenamide failed to prevent vacuolar myopathy after treatment in vivo and failed to inhibit lactate efflux in vitro. CONCLUSIONS AND IMPLICATIONS: Acetazolamide prevents vacuolar myopathy in K(+)-depleted rats. This effect was associated with inhibition of lactate transport, rather than inhibition of CA.

Muscleblind-like protein 1 nuclear sequestration is a molecular pathology marker of DM1 and DM2.

Myotonic dystrophies (DM) are repeat expansion diseases in which expanded CTG (DM1) and CCTG (DM2) repeats cause the disease. Mutant transcripts containing CUG/CCUG repeats are retained in muscle nuclei producing ribonuclear inclusions, which can bind specific RNA-binding proteins, leading to a reduction in their activity. The sequestration of muscleblind-like proteins (MBNLs), a family of alternative splicing factors, appears to be involved in splicing defects characteristic of DM pathologies. To determine whether MBNL1 nuclear sequestration is a feature of DM pathologies, we have examined the in vivo distribution of MBNL1 in muscle sections from genetically confirmed DM1 (n=7) and DM2 (n=9) patients, patients with other myotonic disorders (n=11) and from patients with disorders caused by repeat expansions, but not DM1/DM2 (n=3). The results of our immunofluorescence study indicate that, among patients examined, MBNL1 nuclear sequestration in protein foci is a molecular pathology marker of DM1 and DM2 patients where ribonuclear inclusions of transcripts with expanded CUG/CCUG repeats are also present. These findings indicate that MBNLs might be important targets for therapeutic interventions to correct some of the specific features of DM pathology.

A genomic approach of the hepatitis C virus generates a protein interaction map.

The hepatitis C virus (HCV) causes severe liver disease, including liver cancer. A vaccine preventing HCV infection has not yet been developed, and, given the increasing number of infected people, this virus is now considered a major public-health problem. The HCV genome is a plus-stranded RNA that encodes a single polyprotein processed into at least 10 mature polypeptides. So far, only the interaction between the protease NS3 and its cofactor, NS4A, which is involved in the processing of the non-structural region, has been extensively studied. Our work was aimed at constructing a protein interaction map of HCV. A classical two-hybrid system failed to detect any interactions between mature HCV polypeptides, suggesting incorrect folding, expression or targetting of these proteins. We therefore developed a two-hybrid strategy, based on exhaustive screens of a random genomic HCV library. Using this method, we found known interactions, such as the capsid homodimer and the protease dimer, NS3-NS4A, as well as several novel interactions such as NS4A-NS2. Thus, our results are consistent with the idea that the use of a random genomic HCV library allows the selection of correctly folded viral protein fragments. Interacting domains of the viral polyprotein are identified, opening the possibility of developing specific anti-viral agents, based on their ability to modulate these interactions.

Oculopharyngeal muscular dystrophy in Italy.

Oculopharyngeal muscular dystrophy (OPMD) is an autosomal dominant myopathy particularly frequent in Québec. The few Italian cases thus far described with bilateral ptosis, dysphagia and variable muscle weakness, show non-specific dystrophic findings on muscle biopsies by light microscopy. We describe a 70-year-old Italian woman with an adult-onset ptosis, mild dysphagia and proximal muscle weakness belonging to a family segregating OPMD according to an autosomal dominant mode of inheritance. Clinical features of four of her relatives are reviewed. Muscle biopsy studied by electron microscopy showed the typical 8.5 nm in diameter intranuclear filamentous inclusions (INI). To our knowledge, this is the first Italian report of OPMD with INI. The identification of nuclear inclusions is mandatory in order to confirm the diagnosis prior to linkage analysis.

Neural regulation of acid maltase in an unusual adult onset deficiency.

In a 48-year-old female, the first symptoms apparently manifested themselves 18 years before, with occasional tripping and weakness in both legs. During the next 18 years, weakness progressed and the patient developed a waddling gait; she became unable to rise from a lying or seated position unassisted and the shoulder girdle also became affected. Neurological examination revealed limb and shoulder girdle predominantly involving the lower extremities. We established cell cultures from muscle biopsy specimens obtained from our patient and carried out morphological analysis which, although aspecific, demonstrated clear signs of neurogenic suffering. This was confirmed in EMG studies performed. Biochemical analysis revealed very low acid maltase residual activity. We describe an unusual case of adult-onset acid maltase deficiency (AMD) with neurogenic atrophy and low residual activity. Innervated myofibres prepared by co-culturing the patient's myoblasts, with spinal cord foetal mouse explants were not associated with an abnormal in vitro maturation of the innervated myofibres as expected by the very low residual enzymatic activity found both in the muscle biopsy specimens and in the muscle cultures. There is strong suggestion that factors other than the amount of residual activity must be involved to determine the clinical manifestation of this disease.

Muscle biopsy and cell cultures: potential diagnostic tools in hereditary skeletal muscle channelopathies.

Hereditary muscle channelopathies are caused by dominant mutations in the genes encoding for subunits of muscle voltage-gated ion channels. Point mutations on the human skeletal muscle Na+ channel (Nav1.4) give rise to hyperkalemic periodic paralysis, potassium aggravated myotonia, paramyotonia congenita and hypokalemic periodic paralysis type 2. Point mutations on the human skeletal muscle Ca2+ channel give rise to hypokalemic periodic paralysis and malignant hyperthermia. Point mutations in the human skeletal chloride channel CIC-1 give rise to myotonia congenita. Point mutations in the inwardly rectifying K+ channel Kir2.1 give rise to a syndrome characterized by periodic paralysis, severe cardiac arrhythmias and skeletal alterations (Andersen's syndrome). Involvement of the same ion channel can thus give rise to different phenotypes. In addition, the same mutation can lead to different phenotypes or similar phenotypes can be caused by different mutations on the same or on different channel subtypes. Bearing in mind, the complexity of this field, the growing number of potential channelopathies (such as the myotonic dystrophies), and the time and cost of the genetic procedures, before a biomolecular approach is addressed, it is mandatory to apply strict diagnostic protocols to screen the patients. In this study we propose a protocol to be applied in the diagnosis of the hereditary muscle channelopathies and we demonstrate that muscle biopsy studies and muscle cell cultures may significantly contribute towards the correct diagnosis of the channel involved. DNA-based diagnosis is now a reality for many of the channelopathies. This has obvious genetic counselling, prognostic and therapeutic implications.

Enzymatic activity and morphological differentiation in de novo innervated human muscle cultures.

In the present series of experiments, we studied the effects of developmental neural control on morphological differentiation and on the activity of muscle-specific (creatine kinase, CK; phosphoglycerate mutase, PGAM; phosphorylase, PPL; and phosphofructokinase, PFK) and non-specific (acid maltase, AM; glucose-6-phosphate dehydrogenase, G6PD), human muscle enzymes in de novo innervated muscle cultures. Following innervation of muscle cultures, we noted an increase in the activity of CK, PPL, PFK and AM along with a reduction in G6PD activity. There was also a change of the CK isoenzymes present in the myotubes, i.e. BB and MB are the major isoenzymes in non innervated cultures, but MM becomes predominant following innervation. In the case of PGAM, the only isoenzyme present in the non innervated cultures was BB while the MM isoform appeared only after a prolonged innervation period in most cases--with the exception of AM--these changes in enzyme activity and in the type of isoenzymes present, demonstrate that innervated cultures are more similar to mature muscle. This maturation of enzymatic activity correlates well with the morphological maturation of the myotubes observed following innervation. Such innervated cultures therefore represent a better model with which to study the morphological and biochemical abnormalities associated with neuromuscular diseases.

Cytoplasmic restoration and persistence of glucose-6-phosphate dehydrogenase activity in stable hybrid myotubes.

In previous experiments, we obtained an in vitro restoration of a cytosolic enzyme glucose-6-phosphate dehydrogenase (G6PD) in hybrid myotubes formed between G6PD-deficient and human or murine myoblasts. In the present series of experiments, the degree of restoration was observed and the persistence of the restored activity was evaluated in the hybrids formed. Quantitative measurements of enzymatic activity in single human-human or human-mouse myotubes, identified by using either fluorescent latex microspheres or Hoechst stain 33258, were made by using a computer-controlled Leitz photometer. Histospectrophotometry and statistical analysis showed a 50% restoration of enzymatic activity in human-human hybrids compared to deficient and normal myotubes. The restored activity was uniformly distributed throughout the cytoplasm and persisted in long-term cultures. No nuclear domain was observed for G6PD. Knowledge of the degree of restoration, of the extent of distribution of the products of "competent" nuclei and the demonstration that the correction is not a transitory event in vitro, supports the potential usefulness of myoblast transfer therapy for this type of myopathy.

Biomolecular identification of (CCTG)n mutation in myotonic dystrophy type 2 (DM2) by FISH on muscle biopsy.

Myotonic dystrophy type 2 (DM2) is a dominantly inherited disorder with multisystemic clinical features, caused by a CCTG repeat expansion in intron 1 of the zinc finger protein 9 (ZNF9) gene. The mutant transcripts are retained in the nucleus forming multiple discrete foci also called ribonuclear inclusions. The size and the somatic instability of DM2 expansion complicate the molecular diagnosis of DM2. In our study fluorescence-labeled CAGG-repeat oligonucleotides were hybridized to muscle biopsies to investigate if fluorescence in situ hybridization (FISH), a relatively quick and simple procedure, could be used as a method to diagnose DM2. When FISH was performed with (CAGG)5 probe, nuclear foci of mutant RNA were present in all genetically confirmed DM2 patients (n=17) and absent in all patients with myotonic dystrophy type 1 (DM1; n=5) or with other muscular disease (n=17) used as controls. In contrast, foci were observed both in DM1 and DM2 myonuclei when muscle tissue were hybridized with (CAG)6CA probe indicating that this probe is not specific for DM2 identification. The consistent detection of ribonuclear inclusions in DM2 muscles and their absence in DM1, in agreement with the clinical diagnosis and with leukocyte (CCTG)n expansion, suggests that fluorescence in situ hybridization using (CAGG)5 probes, may be a specific method to distinguish between DM1 and DM2. Moreover, the procedure is simple, and readily applicable in any pathology laboratory.

[Aerospace medicine and its scientific and technological fallout in the field of general biomedicine]. / Medicina aerospaziale e sua "ricaduta" scientifica e tecnologica in campo biomedico generale.

A general preliminary study is made of possible "fall-out" applications from military technology--especially in certain electronics sectors--enabling such technology to be used for non-military purposes that would greatly benefit society, for example in the biomedical field. The continued and profitable exchange that is rapidly developing, not only between technology and science but also between science and technology in the specific sector of aviation medicine, in discussed in detail. The direct and indirect theoretical and practical applications of aviation medicine in medical science, practical medicine and health services are briefly outlined (apparatus for inhaling oxygen; safety and "delethalisation" methods for accidents caused by violent impact; the application of minor technologies in the fields of immunology and hygiene-prophylaxis to air transport and rescue services; the selection and medico-legal check-ups of all military and civilian flight personnel; the emergency transport of transplant organs and seriously ill patients requiring emergency transplants, etc.). Current and possible future practical applications of the vast experience acquired by space medicine had to solve complex problems relating to space flight and man's survival in space, such as: the absence of gravity (and its functional effects on the body's main system and organs), exposure to cosmic and solar radiation, conditioning and prolonged isolation in the space capsule, modifications in circadian rhythms, "space sickness", etc. Finally, the study recalls that the evolution and refinement of aerospace technology has made it possible to make use of exceptional space conditions, such as "microgravity", to conduct in-depth studies--in extreme conditions--of certain biological phenomena and to gain further knowledge of the genesis and to gain of various vital processes. Another area under study is the production--in extremely favourable conditions--of new, vaccines, enzymes and hormones, new drugs for therapy and prophylaxis and for biopharmacology, and new food sources which are of prime importance for man's future requirements.

[Atmospheric pollution. Biological aspects and role of meteorologic factors in the distribution of respiratory hazards in the environment and relative prognostic and preventive measures]. / Inquinamento atmosferico. Aspetti biologici e ruolo dei fattori meteorologici nella distribuzione del rischio respiratorio da ambiente, e relativi mezzi predittivi e preventivi.

The biometeorological factors underlying atmospheric pollution are discussed, together with its biological effects and its direct and indirect damage to human health. Damage is particularly likely when particularly adverse ambient and climatological conditions result in a massive and persistent concentration of contaminants in the ambient air. In this case, since the highly biologically interesting phenomenon of physicochemical reaction between pollutants and atmospheric components prevails over that of the dilution and dispersion of such contaminants, nature's major processes of removal and self-purification may be rendered nugatory and insufficiently prompt. The effects of atmospheric pollution, primarily with respect to urban background pollution, on human health make their appearance in the respiratory system, where there is a continuous relation between man and his environment throughout his life, in the form of immediate or short, medium and long-term damage. The desirability of preparing a meteorological map showing the distribution of the risk of atmospheric pollution is discussed. For this purpose, use could be made of meteorological data, and the hi-tech observations now made possible, inter alia, by the employment of satellites and aerospace data. A map of this kind would give more precise information concerning the part played by the weather in distributing the risk of respiratory damage caused by environmental pollution. It would also provide the knowledge required for the purpose of prediction and prevention in an organised struggle against such pollution. This would be of great social significance and value. Its practical applications could be enormous consequence to humanity.

[Motion sickness in the aerospace environment]. / Le malattie da movimento in ambiente aerospaziale.

After a brief description of the newest and special form of motion sickness known as "space sickness" arising in space flight, and the various hypotheses on its aetiopathogenesis, motion sickness in general and the air or plane sickness deriving from atmospheric flying are discussed. The aetiopathogenesis of air sickness derives from abnormal stimulations that are primarily vestibular but also visual and somesthesic, and generated by irregular movements or variations in attitude of the plane. Reflex action than produces effects that are primarily neurovegetative (nausea, vomiting, pallor, scialorrhea, sweating, bradycardia) and neuropsychological (depression, drowsiness, headache, discomfort and general debility with altered cenesthesia). After a description of the symptoms, the prevention and treatment of air sickness are discussed.

[Evolution and current uses of telematics in medicine. Future prospectives of telemedicine]. / Evoluzione ed applicazioni recenti della telematica in medicina. Prospettive future della telemedicina.

The medical use of data transmission has become a basic element in the application of telecommunications to medical practice. Hence the development of medical telematics i.e. the application of socalled "telematics" derived from the combined use of information science and telecommunications in medicine. In historical terms the foundations for medical telematics were laid in Italy in 1935 with the creation of the International Radiomedical Centre in Rome and more recently with the direct and indirect application of telematics to the flight and medical staff of the Air Rescue Service planes as well as the use--in space medicine--of special sensors computer-recorders, telecameras, etc., for the collection and long-distance transmission of data on the behaviour of men and animals during long space missions over the past 20 years. Other recent techniques enabling the long-distance transmission of data required for the long-distance treatment of the sick are also reviewed with the emphasis on cardiology, neurology, radiology, radiodiagnosis, nuclear medicine, oncology, nephrology, ophthalmology, haematology, laboratory diagnosis, etc. There are many other objectives of supreme human and social value that efficiently organised medical telematics can attain to solve the many difficult problems involved in the gradual development of a truly effective health service for all.

[The Sikorskj HH-3F in rescue activities and in air medical transport]. / Il Sikorskj HH-3F nell'attività di soccorso e trasporto sanitario per via aerea.

Technical and medical problems arising out of air rescue and transport operations are examined through an overview of the activities of the 15th S.A.R. Wing, Italian Air Force. The particular engineering, radio, and S.A.R. features of the Sikorskj HH-3F, currently used for this purpose, make it a sound and effective choice. A specially trained medical, health and nursing unit able to operate on the aircraft under all-weather conditions, coupled with a wide range of medical material, provide the best comment on the series of pathologies illustrated in the tables. Behind this matter, however, there increasingly lies the equally important question of the desirability or otherwise of imposing a strict control to make sure that the use of helicopters on S.A.R. tasks is not indiscriminate, but governed by clearly defined conditions.

Workload and operational fatigue in helicopter pilots.

In light of the modern aetiopathogenic views, a brief review was made concerning possible causes of operational fatigue to which flying personnel in general are exposed in the exercise of flying activity. The author then describes and analyzes the meaning and importance of the various stressing factors that constitute the physical and psychic workload to which the helicopter pilot is subjected in performing his professional activities. Also analyzed are the influences exercised, both separately and jointly, on the genesis of flight fatigue in helicopter pilots by stressing and fatiguing effects of vibrations, noise, and psycho-emotional and psycho-sensorial factors related to the variety and danger of utilization of this modern aircraft. Such an analytical investigation enables the author to conclude that one must admit that helicopter piloting involves a psycho-physical workload certainly no less than that required by more powerful and faster aircraft.

Spinal injury after ejection in jet pilots: mechanism, diagnosis, followup, and prevention.

In order to contribute to the study of spinal injury after ejection., the author analyzed the results of 100 cases of ejections carried out by military and civil Italian jet pilots in a period of 20 years. Of this group, 47 successfully ejected from aircraft without injury; 11 ejections proved fatal. The remaining 42 pilots sutained vertebral fractures, while 27 sustained other traumatic injuries different from spinal fractures. There were 23 vertebral fractures in 15 pilots and the most frequently affected vertebrae were those of the thoraco-lumbar junction. Analysis was make of the pathology, the clinical and radiological profiles, the therapeutic treatment, and the relative aeromedico-legal aspects concerning the temporary unfitness for flying or permanent grounding of the personnel as well as the possible prevention of spinal injury after ejection

New perspectives in the treatment of hypoxic and ischemic brain damage: effect of gangliosides.

Aircrews operating at high G forces and altitudes may be exposed to both physiological and physical stresses capable of inducing brain hypoxia. A potential therapeutic tool for the treatment of flight personnel, monosialoganglioside (GM1) has been found to reduce deficits and enhance repair following CNS injury. A survey of experimental evidence concerning the effects of GM1 in the acute phase of CNS injury supports its proposed application for aerospace medicine.

Biofeedback monitoring-devices for astronauts in space environment.

After a reconsideration of the state-of-the-art in biofeedback research the implementation of biofeedback systems is envisioned as a countermeasure of stress for the psychoprophylaxis of the astronaut. A one-session experiment performed on two groups of subjects to assess the interference from EMG-feedback on the performance in a simultaneous psychomotor trial with a view to expanding biofeedback application is described. The results show that the experimental group performed in the same way as the control without feedback, but with less CNS activation. Some general conclusions are drawn from the advances in technology.

Italy's contribution, from a medical standpoint, to the space safety of payload scientists, and perspectives for the future.

In Italy, the selection of the Italian payload scientists has been performed according to the Spacelab Program of ESA. Twenty-four subjects underwent a screening performed by the Health Service of Italian Air Force. They were requested to pass an exercise test on treadmill and another ten-minute test on centrifuge, subject to the effect of +3 Gz. The authors briefly describe the results of the test. Noteworthy is the determination of Central Flicker Fusion Frequency. This parameter makes it possible to assess the endurance level of the subject, much earlier than other techniques (e.g. EKG). The importance of an accurate preliminary screening is emphasized as well as of successive training periods. Future studies will be undertaken to compare evoked cortical potentials with behaviour parameters of space safety, with a view to setting up a subtle tool of evaluation for both future candidates and payload scientists.