Plasma heavy metal levels correlate with deregulated gene expression of detoxifying enzymes in osteoporotic patients.

Heavy metal levels appear to be associated with low bone mineral density (BMD) and the consequent osteoporosis risk, but the relationship with the disease has not been clearly defined. The altered expression pattern of numerous genes, including detoxifying genes, seems to play a pivotal role in this context, leading to increased susceptibility to several diseases, including osteoporosis. The purpose of this study is to analyse circulating heavy metals levels and the expression of detoxifying genes in osteoporotic patients (OPs, n = 31), compared with healthy subjects (CTRs, n = 32). Heavy metals concentration in plasma samples was determined by Inductively Coupled Plasma Mass Spectrometry (ICP-MS), and the subsequent expression analysis of NAD(P)H quinone dehydrogenase 1 (NQO1), Catalase (CAT), and Metallothionein 1E (MT1E) genes in Peripheral Blood Mononuclear Cells (PBMCs) was assessed by real-time polymerase chain reaction (qRT-PCR). Copper (Cu), mercury (Hg), molybdenum (Mo) and lead (Pb) were found to be significantly higher in the plasma of OPs compared to CTRs. Analysis of the expression levels of detoxifying genes showed a significant decrease in CAT and MT1E in OP group. In addition, Cu correlated positively with the expression levels of both CAT and MT1E in CTRs group and MT1E in OPs. This study shows an increased circulating concentration of certain metals combined with an altered expression pattern of detoxifying genes in OPs, highlighting a novel aspect to be investigated in order to better characterize the role of metals in the pathogenesis of osteoporosis.

Risk factors for sporadic Campylobacter infection: an all-Ireland case-control study.

We report the findings of the first case-control study conducted in both the Republic of Ireland and Northern Ireland to determine risk factors for sporadic Campylobacter infections. A total of 197 cases and 296 case-nominated controls matched for age, were included. Based on Population Attributable Fraction (PAF), the most important risk factors were consuming chicken [adjusted matched (am) OR 6.8; 95%CI 2.1-21.9], consuming lettuce (amOR 3.3; 95%CI 1.5-7.1) and eating in takeaways (amOR=3.1; 95%CI 1.4-6.6). Contact with sheep (amOR=11; 95%CI 1.6-78), peptic ulcer (amOR=19; 95%CI 3.8-93.7), hiatus hernia (amOR=20.3; 95%CI 2.3-183.3), lower bowel problems (amOR=4.5; 95%CI 1.2-16.8) were also independently associated with infection. Mains water supply showed protective effect (amOR=0.2; 95 CI 0.1-0.9). The findings highlight the continued need for consumer food safety education and further control measures throughout the food chain on the island of Ireland.

Olfactory sensory axons expressing a dominant-negative semaphorin receptor enter the CNS early and overshoot their target.

Sensory axons extend from the chick olfactory epithelium to the telencephalon well before the maturation of their target, the olfactory bulb. During a waiting period of several days, olfactory axons arrive and accumulate outside the CNS while the bulb differentiates beneath them. Semephorin-3A is expressed in the tel-encephalon during this period and has been proposed to prevent their entry into the CNS. We show that the misexpression of a dominant-negative neuropilin-1 that blocks SEMA-3A-mediated signaling in olfactory sensory axons induces many of them to enter the tel-encephalon prematurely and to overshoot the olfactory bulb. These results suggest that chemorepellents can prevent the premature innervation of immature targets.

A family of molecules related to collapsin in the embryonic chick nervous system.

Signaling molecules with either attractive or repulsive effects on specific growth cones are likely to play a role in guiding axons to their appropriate targets. A chick brain glycoprotein, collapsin, has been shown to be a good candidate for a repulsive guidance cue. We report here the discovery of four new molecules related to collapsin in chick brains. All contain a semaphorin domain. One is structurally very similar to collapsin but is only 50% identical in its amino acid sequence. We have named it collapsin-2. The collapsin-related genes exhibit distinct but overlapping patterns of mRNA expression in the developing spinal cord and the developing visual system. This family of collapsin-related molecules could potentially act as repulsive cues toward specific neuronal populations.

Post-ischemic treatment with erythropoietin or carbamylated erythropoietin reduces infarction and improves neurological outcome in a rat model of focal cerebral ischemia.

BACKGROUND AND PURPOSE: Recombinant human erythropoietin (rhEPO; Epoetin-alpha; PROCRITtrade mark) has been shown to exert neuroprotective and restorative effects in a variety of CNS injury models. However, limited information is available regarding the dose levels required for these beneficial effects or the neuronal responses that may underlie them. Here we have investigated the dose-response to rhEPO and compared the effects of rhEPO with those of carbamylated rhEPO (CEPO) in a model of cerebral stroke in rats. EXPERIMENTAL APPROACH: Rats subjected to embolic middle cerebral artery occlusion (MCAo) were treated with rhEPO or CEPO, starting at 6 h and repeated at 24 and 48 h, after MCAo. Cerebral infarct volumes were assessed at 28 days and neurological impairment at 7, 14, 21 and 28 days, post-MCAo. KEY RESULTS: rhEPO at dose levels of 500, 1150 or 5000 IU kg(-1) or CEPO at a dose level of 50 microg kg(-1) significantly reduced cortical infarct volume and reduced neurologic impairment. All doses of rhEPO, but not CEPO, produced a transient increase in haematocrit, while rhEPO and CEPO substantially reduced the number of apoptotic cells and activated microglia in the ischemic boundary region. CONCLUSIONS AND IMPLICATIONS: These data indicate that rhEPO and CEPO have anti-inflammatory and anti-apoptotic effects, even with administration at 6 h following embolic MCAo in rats. Taken together, these actions of rhEPO and CEPO are likely to contribute to their reduction of neurologic impairment following cerebral ischemia.

Assessment of Salmonella survival in dry-cured Italian salami.

The inactivation of Salmonella during curing of Italian traditional pork salami was investigated. A total of 150 batches of ground raw meat (GRM) used for salami manufacturing by four producers were tested for Salmonella by real-time PCR followed by ISO 6579 cultural confirmation and MPN enumeration. Salami produced with Salmonella positive GRMs were re-tested at the end of their curing period. Aw, pH and NaCl content were also measured. Detection of Salmonella was performed testing both 25 and 50g of the samples. By Real-Time PCR 37% of the GRMs resulted positive, but cultural detection of Salmonella was obtained in 14% of the samples only. Salmonella enumeration ranged from 31 MPN/g to <1.3 MPN/g. The difference between testing 50g and 25g of the samples was statistically significant (p value≤0.01). In particular, ISO-50g detected Salmonella in 100% of all positive samples, vs. 62% of ISO-25g. Salami made of the contaminated GRMs were 29% Salmonella-positive, as most batches of salami produced with Salmonella-positive GRMs resulted negative after regular curing (20-48days). Overall, 13% of salami produced with Salmonella-contaminated GRMs were positive. They belonged to six batches, which turned out negative after prolonged curing ranging between 49 and 86days. Salmonella enumeration in salami ranged from 8.7 MPN/g to <1.3 MPN/g. Unlike GRMs, no significant difference was observed between the ISO-50g and the ISO-25g in detecting Salmonella in cured salami (p value: >0.05). The most common Salmonella serovars in GRMs were Derby (52%), Typhimurium monophasic variant 4, (Barbuti et al., 1993), 12:i:- (19%) and Stanley (10%). Salmonella Derby (56%), London, Branderup, Panama (13%, respectively) and Goldcoast (6%) were most frequent in cured salami. The study showed negative correlation between real-time CT values and cultural confirmation of Salmonella, as well as the importance of sample size for Salmonella detection. Among considered factors with possible effect on the occurrence of Salmonella in salami, statistical analysis revealed a role for aw in salami and for Salmonella load in GRMs, while pH and NaCl content did not significantly affect the probability of finding Salmonella in dry-cured salami in the context of this study. In particular the lower aw values due to longer curing were associated with lower Salmonella presence in traditional dry-cured salami.

A dominant negative receptor for specific secreted semaphorins is generated by deleting an extracellular domain from neuropilin-1.

Neuropilins have recently been characterized as receptors for secreted semaphorins. Here, we report the generation of a dominant negative form of neuropilin-1 by the deletion of one of its extracellular domains. Expression of this variant in cultured primary sympathetic neurons blocks the paralysis of growth cone motility normally induced by SEMA-3A (collapsin-1, semaphorin III, semaphorin D) and SEMA-3C (collapsin-3, semaphorin E) but not that induced by SEMA-3F (semaphorin IV). A truncated form of neuropilin-1 that is missing its cytoplasmic domain fails to act as a dominant negative receptor component. These results suggest that neuropilin-1 is a necessary component of receptor complexes for some, but not all, secreted semaphorin family members. Overexpression of dominant negative neuropilins should provide a powerful new method of blocking the functions of secreted semaphorins.

Transesophageal dipyridamole echocardiography for diagnosis of coronary artery disease.

The value of transthoracic dipyridamole echocardiography has been extensively documented. However, in some patients, because of a poor acoustic window, the rest transthoracic examination is not always feasible and the transesophageal approach is more convenient. Therefore, transesophageal echocardiography with high dose dipyridamole (up to 0.84 mg/kg body weight over 10 min) was performed in 32 patients in whom the transthoracic dipyridamole test either was not feasible (n = 29) or yielded ambiguous results (n = 3). The transesophageal echocardiographic test results were considered abnormal when new dipyridamole-induced regional wall motion abnormalities were observed. All 32 patients underwent coronary angiography; significant coronary artery disease was defined as greater than or equal to 70% lumen diameter narrowing in at least one major vessel. All patients also performed a bicycle exercise test 1 day before transesophageal dipyridamole echocardiography. Transesophageal stress studies were completed in all patients, with a maximal imaging time (in tests with a negative result) of 20 min. No side effects or intolerance to drug or transducer was observed. The left ventricle was always visualized in the four-chamber and transgastric short-axis views. High quality two-dimensional echocardiographic images were obtained in all patients both at rest and at peak dipyridamole infusion and were digitally analyzed in a quad-screen format. Coronary angiography showed coronary artery obstruction in 24 patients: 6 had single-, 9 double- and 9 triple-vessel disease. The transesophageal dipyridamole test showed a specificity of 100% and an overall sensitivity of 92%. The sensitivity of this test for single-, double- and triple-vessel disease was 67%, 100% and 100%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Sugar as a vehicle for iron fortification: further studies.

The data presented confirm the advantages of sugar as a vehicle for iron fortification over other vehicles used in the past. The absorption comparison between ferric and ferrous salts added to sugar demonstrated that Fe(III)-EDTA Complex and ferrous sulfate exhibited the highest absorption, while ferric ammonium citrate was poorly absorbed. It was also found that Fe(III)-EDTA reacts slowly with the tannin contained in tea; the color of the tea changes slightly in the first 2 hr after the addition of the fortified sugar. Iron absorption of sugar fortified with ferrous sulfate was tested in seven beverages. The mean absorption ratio from fortified sugar given with beverages to reference dose of iron ascorbate ranged between 0.42 and 0.70, that is, more than 4 times the absorption from fortified sugar when it is administered with a meal containing one or more vegetals. An absorption of between 0.25 and 0.80 mg of iron/soft drink sugar fortified with 3 mg of iron as ferrous sulfate can be expected in subjects with various degrees of iron deficiency. Thus, two soft drinks per day between meals would be enough to meet the iron requirement in more than 95% of menstruating women, even though the daily iron absorption from the diet is about 0.8 to 1.0 mg.

Fe(III)-EDTA complex as iron fortification. Further studies.

The data presented confirm the advantages of Fe(III)-EDTA as a salt for iron fortification. This iron compound exchanges completely with intrinsic wheat iron in the lumen of the gut. The iron absorption data from this salt tested with six different food vehicles compared with the absorption of ferrous sulfate administered with the same vehicles indicate that while the mean absorption from ferrous sulfate varies from 2 to 30% according to the food vehicle mixed with the salt, the absorption from Fe(III)-EDTA remains practically the same. Apparently, the iron absorption from Fe(III)-EDTA complex is slightly or not affected by the presence of vegetable foods or milk. All these data suggest that only a small amount of iron from this salt, about 10 mg/day, would be necessary to prevent iron deficiency anemia even in those populations relying for their subsistence on vegetable food only.

Sugar as a vehicle for iron fortification.

Sugar as a vehicle for iron fortification presents several advantages over the other vehicles used in the last three decades. In vitro studies demonstrated that ferrous sulfate added to sugar in proportion of 1 mg to 1 g, respectively, is maintained in the ferrous form for a period of at least 1 year and does not induce adverse changes in the vehicle. Sugar, by itself, carries practically no inhibitors for the absorption of iron. Iron absorption from fortified sugar mixed with vegetals is the same as that of native vegetal iron. The absorption from fortified sugar is increased more than 50% over that observed from native vegetal when it is administered as a drink during the ingestion of a meal. A further increase in absorption was found when fortified sugar was administered with beverages. The mean absorption ratio of fortified sugar given with orange juice, Coca-Cola, and Pepsi-Cola to a reference dose of iron ascorbate was between 0.45 and 0.66, which is more than 3 times the absorption of this iron fortification mixed with vegetals. The mean absorption ratio from coffee was 0.30, and from coffee with milk, 0.15. These data indicate that the fortification of sugar with iron could be a better procedure for the prevention of iron deficiency than the iron fortification of bread and wheat products, from which iron is poorly absorbed. It could be used in developing countries where beverages are highly consumed by the low socioeconomic class. This program could be extended to all sugar consumption or be restricted to soft drinks.

Effects of quisqualic acid analogs on metabotropic glutamate receptors coupled to phosphoinositide hydrolysis in rat hippocampus.

L-Glutamic acid (L-Glu) and L-aspartic acid (L-Asp) activate several receptor subtypes, including metabotropic Glu receptors coupled to phosphoinositide (PI) hydrolysis. Quisqualic acid (Quis) is the most potent agonist of these receptors. There is evidence that activation of these receptors may cause a long lasting sensitization of neurons to depolarization, a phenomenon called the Quis effect. The purpose of the current studies was to use Quis analogs and the recently identified metabotropic receptor antagonist, (+)-alpha-methyl-4-carboxy-phenylglycine((+)-MCPG), to define the structural properties required for interaction with the metabotropic receptors coupled to PI hydrolysis and to determine if the Quis effect is mediated by these receptors. The effects of Quis analogs on PI hydrolysis were studied in the absence or presence of the metabotropic receptor-specific agonist 1SR,3RS-1-amino-1,3-cyclopentanedicarboxylic acid (1SR,3RS-ACPD) in neonatal rat hippocampus. Some of the compounds that induce the Quis effect also stimulate PI hydrolysis, including Quis itself and 9 (homoquisqualic acid). Not all of the Quis analogs that stimulate PI hydrolysis, however, induce the Quis effect, including 7A (EC50 = 750 +/- 150 microM) and (RS)-4-bromohomoibotenic acid (BrHI) (EC50 = 130 +/- 40 microM). Although (+)-MCPG blocked PI hydrolysis stimulated by Quis (IC50 = 370 +/- 70 microM), it had no effect on the induction of the Quis effect. Other Quis analogs did not stimulate PI hydrolysis but rather blocked the effects of 1SR,3RS-ACPD. The IC50 values were 240 +/- 70 microM for 2, 250 +/- 90 microM for 3, and 640 +/- 200 microM for 4. Data for inhibition by 2 and 3 were consistent with non-competitive mechanisms of action. These studies provide new information about the structural features of Quis required for interaction with metabotropic receptors coupled to PI hydrolysis and provide evidence that the Quis effect is not mediated by (+)-MCPG sensitive subtypes of these receptors.

N-aryl-N'-benzylpiperazines as potential antipsychotic agents.

N1-(2-Alkoxyphenyl)piperazines additionally containing an N4-benzyl group bearing alcohol, amide, imide, or hydantoin functionalities were prepared and evaluated in the conditioned avoidance response (CAR) test predictive of clinical antipsychotic activity and in in vitro receptor-binding assays. Certain of the compounds display high affinity for the D2, 5-HT1A, and alpha 1-adrenergic receptors. Structures bearing acyclic amide, lactam, and imide functionalities display good biological activity, with a preference for the 1,3-disubstituted phenyl ring relative to the 1,4- and 1,2-congeners (7 vs 10 and 12). Every possible position of hydantoin attachment was investigated (e.g., substitution at N1, N3, and C5). The hydantoin involving attachment to N1 (24) was found to have good biological activity, whereas those hydantoins with attachment to N3 or C5 (22, 23, and 25) were inactive. Several of the smaller acetylated derivatives (30 and 33) have fair in vivo activity, which was lost in the case of the larger benzoyl analog 31. Uracil congener 34 had modest affinity for the D2 receptor (65 nM) as well as excellent in vivo activity. Benzylamino compounds display (viz. 27 and 35-38) moderate CAR activity but have surprising receptor affinity, often greater than those of comparable structures bearing a carbonyl (36 vs 7). Benzyl and benzhydryl alcohol compounds 40-48 are more active than amino structures 27 and 35-38 and also exhibit excellent in vivo activity in the CAR test with modest D2 and 5-HT1A receptor binding.

Orally active benzamide antipsychotic agents with affinity for dopamine D2, serotonin 5-HT1A, and adrenergic alpha1 receptors.

New antipsychotic drugs are needed because current therapy is ineffective for many schizophrenics and because treatment is often accompanied by extrapyramidal symptoms and dyskinesias. This paper describes the design, synthesis, and evaluation of a series of related (aminomethyl)benzamides in assays predictive of antipsychotic activity in humans. These compounds had notable affinity for dopamine D2, serotonin 5-HT1A, and alpha1-adrenergic receptors. The arylpiperazine 1-[3-[[4-[2-(1-methylethoxy)phenyl]-1-piperazinyl]methyl]benzoyl]p ipe ridine (mazapertine, 6) was chosen because of its overall profile for evaluation in human clinical trials. The corresponding 4-arylpiperidine derivative 67 was also highly active indicating that the aniline nitrogen of 6 is not required for activity. Other particularly active structures include homopiperidine amide 14 and N-methylcyclohexylamide 31.

Chemokine receptor CXCR2 regulates the functional properties of AMPA-type glutamate receptor GluR1 in HEK cells.

Experiments were conducted in both HEK cells and cerebellar neurons to investigate whether CXC chemokine receptor 2 (CXCR2) is functionally coupled to GluR1. The co-expression of CXCR2 with GluR1 in HEK cells increased (i) the GluR1 "apparent" affinity for the transmitter; (ii) the GluR1 channel open probability; and (iii) GluR1 binding site cooperativity upon CXCR2 stimulation with CXC chemokine ligand 2 (CXCL2). The affinity of C-terminal-deleted GluR1 for glutamate (Glu) remained stable instead. Furthermore, CXCL2 increased the binding site cooperativity of AMPA receptors in rat cerebellar granule cells; and the amplitude of spontaneous excitatory postsynaptic current (sEPSCs) in Purkinje neurons (PNs). Our findings indicate that the coupling of CXCR2 with GluR1 may modulate glutamatergic synaptic transmission.

Nicotinic cholinergic stimulation promotes survival and reduces motility of cultured rat cerebellar granule cells.

Despite many studies on the functional expression of neuronal nicotinic acetylcholine receptors (nAChRs), an exhaustive description of the long-term effects of nicotine (Nic) stimulation in cerebellar granules is still far to be completed. For this reason, we addressed the experiments stimulating cultured cerebellar granule neurons (CGN) with Nic, focusing on the effects on cell motility and survival. Using electrophysiological and Ca(2+)-fluorescence techniques, we found a subset of rat CGN that responded to Nic by inward whole cell currents and by short-delay Ca(2+) transients. These responses were mediated through both homomeric and heteromeric nAChRs, as assessed by their sensitivity to alpha-bungarotoxin (alpha-BTX), dihydro-beta-erythroidine (DHbetaE), methyllicaconitine (MLA) and 5-hydroxyindole (5OH-indole). Once established the expression of alpha-BTX-sensitive and insensitive nAChRs and their ability to trigger Ca(2+) responses in CGN, we aimed at investigating their possible role on cell survival and motility. We demonstrate that Nic stimulation significantly increases the survival of CGN exposed to the apoptosis-promoting low K(+) medium. This anti-apoptotic effect is likely mediated through alpha7* nAChRs since we found that it was mimicked by choline, was insensitive to DHbetaE and was fully inhibited by alpha-BTX. Furthermore, we report that Nic negatively modulates CGN motility, reducing the basal cell movement through a pored membrane by the activation of alpha-BTX-insensitive nAChRs. We conclude that CGN express various types of nAChRs, which are differently involved in regulating Nic-mediated modulation of cell survival and migration, and we suggest potential regulatory roles for cholinergic receptors during cerebellar development.

Assessment of severity of coronary narrowings by quantitative exercise echocardiography and comparison with quantitative arteriography.

To determine the correlation of quantitative assessment of coronary narrowings with left ventricular functional impairment induced by exercise, 57 patients with 1-vessel coronary artery disease and without evidence of collateral flow were studied. A significant relation was observed between minimal cross-sectional area, percent area stenosis, minimal lumen diameter, percent diameter stenosis and the percentage of segmental area change from rest to peak exercise in a vascular distribution territory (r = 0.76, p less than 0.001; r = -0.55, p less than 0.001; r = 0.56, p less than 0.001; r = -0.75, p less than 0.001, respectively). For minimal cross-sectional area, the best cut-off value to separate significantly patients who had a decrease in contractility at peak exercise testing from those who had a normal response was 2 mm2 (p less than 0.001); for percent cross-sectional area stenosis, it was 75% (p less than 0.001); for minimal lumen diameter, it was 0.7 mm (p less than 0.001); and, for percent diameter stenosis, it was 85% (p less than 0.001). High cut-off values for angiographic variables are necessary to separate significantly patients who have a decrease in contractility at peak exercise testing from those who have a normal response. Several patients with mild coronary stenoses may have either normal or abnormal wall motion during exercise. Thus, exercise echocardiography is a useful tool in detecting the presence of fairly severe anatomic narrowing, whereas it is of limited clinical use in the assessment of intermediate coronary atherosclerotic lesions.

Epidemiology and current situation of viral haemorrhagic disease of rabbits and the European brown hare syndrome in Italy.

Rabbit production is of considerable economic importance in Italy. In the last thirty years, meat production has risen and the number of intensive husbandry establishments has grown. The major region of production (about 60%) lies in the northern part of the country. In addition, approximately one million live animals and more than 14,000 tons of meat are imported yearly. More than 150,000 hares are also imported for restocking and hunting purposes. During the second half of the 1980s a "new" disease (X disease), characterised by haemorrhagic and degenerative lesions and high mortality, was observed on traditional farms in some regions. This form spread throughout the country between 1986 and 1988. In 1988, diagnostic research led to identification of a viral agent. Because of its morphology, the virus was at first considered a picornavirus. More recent research has included the virus in the Caliciviridae family. This agent reproduces the disease experimentally. Between 1988 and 1989, many intensive establishments in several regions were affected by the disease, known as viral haemorrhagic disease (VHD); the losses were always very high. In 1989, after mortality among free-living hares had been observed in the northern part of Italy for some years, the agent of the European brown hare syndrome (EBHS) was identified as being morphologically similar to the VHD virus. The antigenic relationship between VHD and EBHS and the possibility of cross-infection between rabbits and hares are currently being studied. Hygienic measures and vaccination are the most reliable methods of control. International cooperation has greatly helped in the study of the disease and in the adoption of control measures.

[Colorimetric and gas chromatographic methods of determination of the biotin in pharmaceutical preparations]. / Metodo colorimetrico e gascromatografico per la determinazione della biotina nelle preparazioni farmaceutiche.

An analytical method for biotin is described, which is based on its conversion to N-nitrosobiotin, purifications of this derivative and its colorimetric or gas chromatographic determination. Nitrosation is carried out by treatment with an excess of NaNO2 in 0.2 N H2SO4 at room temperature. Nitrosobiotin is extracted with chloroform, after salting out wiulphanilamide + HCl), in order to obtain hydrolysis, and then with the second oating conditions are:a glass column, 2 m long, packed with 3% SE-30 on Chromosorb WHP; colum temperature 190 degrees C; injector and detector temperature 260 degrees C; carrier gas: nitrogen (30 ml/min). The extraction of biotin from multivation solid preparations is performed with absolute ethanol, starting from an expected quantity of about 250 microgram. It is suggested to work at the same time with an internal standard.

[Minimally-invasive endoscopic transthoracic sympathectomy of the upper limbs. A new method]. / Denervazione simpatica mini-invasiva degli arti superiori. Nuova metodica.

OBJECTIVE: Indications for endoscopic transthoracic upper dorsal sympathectomy are axillary and palmar hyperhidrosis, upper extremities ischemia (due to, e.g., Raynaud s disease), and upper extremities causalgia. METHODS: At present, this methodology relies on (at least) double trocar insertion (per side) and/or carbon dioxide insufflation. Thus, although this approach, compared with the traditional open sympathectomy techniques, it guarantees the smallest number of postoperative complications, it still determines a certain amount of postoperative discomfort as well as a risk of complications related to carbon dioxide insufflation, as intraoperative profound bradycardia and hypotension due to mediastinal shift, and postoperative subcutaneous emphysema. From December 1995, we are using a minimally-invasive endoscopic transthoracic sympathectomy technique, performed by a single-entry specifically modified thoracoscope and without the need for carbon dioxide insufflation, with the aim to reduce the drawbacks associated with the above-mentioned currently adopted endoscopic techniques. After general anesthesia with double-lumen endotracheal tube, with the patient placed in a half-sitting position with both arms abduced to 90 degrees, a 1 cm incision is performed, along the midclavear line (in male patients) or the anterior axillary line (in female patients), in the second or third intercostal space. RESULTS: The effects of sympathectomy are immediate, and the patients wake up with warm and dry hands and axillae. CONCLUSIONS: In personal opinion, this single-entry technique, compared with other reported approaches, should minimize any damage to the intercostal neurovascular bundle, while avoiding the complications connected with carbon dioxide insufflation.