[Permanent environmental disinfection techniques in hospital settings with infectious risk]. / Tecniche di disinfezione ambientale permanente negli ambienti ospedalieri a rischio infettivo.

While the world's economies avoid the COVID-19 pandemic blockade, there is an urgent need for technologies aimed at reducing the transmission of COVID-19 in confined spaces such as hospital environments. Although the cleaning and disinfection procedures now have rather complex and sophisticated weapons, they do not seem to be sufficient to continuously maintain low levels of environmental microbiological contamination. This result can now be achieved through the cross-use, in space and time, of improved, more efficient and effective technologies. This result can now be achieved through the cross-use, in space and time, of improved technologies. This work highlights the possibility of crossing and cooperation of different disinfection techniques, such as to keep the microbial and viral load low over time.

Binding, internalization and fate of Huntingtin Exon1 fibrillar assemblies in mitotic and nonmitotic neuroblastoma cells.

AIMS: The aggregation of Huntingtin (HTT) protein and of its moiety encoded by its Exon1 (HTTExon1) into fibrillar structures inside neurons is the molecular hallmark of Huntington's disease. Prion-like transmission of these aggregates between cells has been demonstrated. The cell-to-cell transmission mechanisms of these protein aggregates and the susceptibility of different kinds of neuronal cells to these toxic assemblies still need assessment. METHODS: Here, we documented the binding to and internalization by differentiated and undifferentiated neuroblastoma cells of exogenous fibrillar HTTExon1 and polyglutamine (polyQ) polypeptides containing the same number of glutamines. We assessed the contribution of endocytosis to fibrillar HTTExon1 uptake, their intracellular localization and fate. RESULTS: We observed that undifferentiated neuroblastoma cells were more susceptible to fibrillar HTTExon1 and polyQ than their differentiated counterparts. Furthermore, we demonstrated that exogenous HTTExon1 aggregates are mainly taken up by endocytosis and directed to lysosomal compartments in both mitotic and quiescent cells. CONCLUSIONS: These data suggest that the rates of endocytic processes that differ in mitotic and quiescent cells strongly impact the uptake of exogenous HTTExon1 and polyQ fibrils. This may be either the consequence of distinct metabolisms or distributions of specific protein partners for amyloid-like assemblies at the surface of highly dividing versus quiescent cells. Our results highlight the importance of endocytic processes in the internalization of exogenous HTTExon1 fibrils and suggest that a proportion of those assemblies reach the cytosol where they can amplify by recruiting the endogenous protein after escaping, by yet an unknown process, from the endo-lysosomal compartments.

Treatment of a bone defect consequent to the removal of a periapical cyst with equine bone and equine membranes: clinical and histological outcome.

AIM: While deproteinized bovine bone and bovine membranes have been well studied and can yield good results when used to treat bone defects and peri-implant dehiscences, enzymatically deantigenated equine bone and equine membranes have emerged as possible alternative biomaterials. The objective of this study was the clinical and histological assessment of such materials: equine bone granules, an equine collagen membrane and an equine pericardium membrane. METHODS: Enzymatically deantigenated equine bone and an equine collagen membrane were used to restore a bone defect caused by the removal of a bone cyst in the upper anterior maxilla. After 4.5 months, an implant was placed and a bone core sample was obtained from the grafted site. Implants threads, though, were exposed. This defect was grafted with a mixture of autogenous and equine bone and covered with an equine pericardium membrane. RESULTS: Four months after implant placement the peri-implant bone levels were maintained. A prosthesis was delivered three months later providing functional and esthetic rehabilitation. Also four-year follow-up controls showed implant success. Histological analysis of the bone core revealed that the graft material had undergone remodelling, and a fair amount of newly formed vital bone was present at the time of sample collection. CONCLUSION: The deantigenated equine bone is biocompatible and undergoes osteoclastic remodelling. Both the equine collagen and pericardium membrane acted as effective barriers for guided bone regeneration.

Prevalence of multifocal atherosclerosis and comorbidity on symptomatic Cypriot inpatients.

AIM: Multifocal atherosclerotic disease or multifocal ischemic pathology (MAD) is an issue gaining a lot of attention by clinicians in recent years, due to its high impact on the morbidity and mortality of vascular patients. The coexistence of coronary artery disease (CAD), peripheral arterial disease (PAD) and carotid disease (CS) is being investigated in this study. METHODS: The study included 556 consecutive inpatients who were admitted to the Nicosia General Hospital in Cyprus for carotid endarterectomy, peripheral arterial reconstruction or coronary artery bypass. All patients were subjected to color duplex examination of the carotid vessels and to lower extremity Doppler study. Cardiac risk screening was performed on patients with PAD and CS. Comorbidity was evaluated by using the Cumulative Illness Rating Scale (CIRS). RESULTS: The prevalence of MAD in atherosclerotic patients was found very high (60.3%). The most frequent comorbidity was the coexistence of PAD and CAD (41.8%). The comorbidity burden of MAD patients, in comparison to that of patients with unifocal atherosclerotic disease, was evaluated to conclude that the burden in the first group (MAD) is significantly higher, not only in relation to the number of clinically relevant diseases that co-exist, but also to the severity of these diseases. Furthermore, functional status is negatively affected by the existence of multifocal ischemic pathology. CONCLUSION: The high prevalence of MAD suggests the necessity of developing a systematic screening approach in the everyday practice. Apart from cardiovascular problems, other diseases affect the clinical situation of these patients. Therefore, it is important to investigate these problems pre- and postoperatively.

Splanchnic vein thrombosis in myeloproliferative neoplasms: risk factors for recurrences in a cohort of 181 patients.

We retrospectively studied 181 patients with polycythaemia vera (n=67), essential thrombocythaemia (n=67) or primary myelofibrosis (n=47), who presented a first episode of splanchnic vein thrombosis (SVT). Budd-Chiari syndrome (BCS) and portal vein thrombosis were diagnosed in 31 (17.1%) and 109 (60.3%) patients, respectively; isolated thrombosis of the mesenteric or splenic veins was detected in 18 and 23 cases, respectively. After this index event, the patients were followed for 735 patient years (pt-years) and experienced 31 recurrences corresponding to an incidence rate of 4.2 per 100 pt-years. Factors associated with a significantly higher risk of recurrence were BCS (hazard ratio (HR): 3.03), history of previous thrombosis (HR: 3.62), splenomegaly (HR: 2.66) and leukocytosis (HR: 2.8). Vitamin K-antagonists (VKA) were prescribed in 85% of patients and the recurrence rate was 3.9 per 100 pt-years, whereas in the small fraction (15%) not receiving VKA more recurrences (7.2 per 100 pt-years) were reported. Intracranial and extracranial major bleeding was recorded mainly in patients on VKA and the corresponding rate was 2.0 per 100 pt-years. In conclusion, despite anticoagulation treatment, the recurrence rate after SVT in myeloproliferative neoplasms is high and suggests the exploration of new avenues of secondary prophylaxis with new antithrombotic drugs and JAK-2 inhibitors.

L-cycloserine: behavioural and biochemical effects after single and repeated administration to mice, rats and cats.

L-Cycloserine dose-dependently inhibited the activity of gamma-aminobutyric acid (GABA)-transaminase (GABA-T) and elevated the level of GABA in whole mouse brain with a peak effect 3-4 hr after a single intraperitoneal injection. At a dose (30 mg/kg) which elevated the level of GABA almost 4-fold, L-cycloserine moderately increased the content of alanine and slightly reduced that of aspartate, glutamate and glycine in the brain. L-Cycloserine (10-30 mg/kg, p.o. or i.p.) prevented tonic seizures induced by 3-mercaptopropionic acid (3-MPA) and audiogenic seizures in DBA/2 mice, without affecting those evoked by pentylenetetrazol, bicuculline and electroshock. Similarly small doses of L-cycloserine reduced the level of cGMP in the cerebellum of rats, prevented its elevation by 3-MPA and attenuated the hypothalamically-elicited rage reaction in cats. Larger doses of L-cycloserine (greater than 30-100 mg/kg) impaired the performance of mice in the rotarod, chimney and horizontal wire tests, and reduced spontaneous locomotor activity of rats. Upon repeated administration the inhibitory effect of L-cycloserine on the activity of GABA-T and on seizures elicited by 3-MPA in mice increased. In contrast, the depressant action of L-cycloserine on motor performance and locomotion declined in subchronically-treated mice and rats. The levels of amino acids in brain after repeated administration did not differ markedly from those in acutely-treated mice. It is suggested that small doses of L-cycloserine, probably by increasing GABAergic inhibition, reduce hyperexcitability in the brain in acute- and subchronically-treated animals. Larger doses of L-cycloserine, possibly by inducing multiple neurochemical changes, evoke central depressant effects which diminish during subchronic treatment.

In vitro and in vivo evaluation of iodine-123-Ro 16-0154: a new imaging agent for SPECT investigations of benzodiazepine receptors.

The flumazenil analogue, Ro 16-0154, a benzodiazepine partial inverse agonist, has been labeled by halogen exchange to enable SPECT investigations of central benzodiazepine receptors in the human brain. The purified 123I-Ro 16-0154 was found to be stable in rat brain preparations and to be metabolized in rat liver preparations. Its pharmacologic properties were comparable to those of flumazenil. The biodistribution in rats (1 hr postinjection) resulted in a high brain-to-blood ratio of 16. Clinical studies revealed images of the benzodiazepine receptor density in the brain. Since the receptor labeling was markedly reduced by injection of flumazenil, it was considered to be specific. Storage defects due to pathologic cerebral blood flow and changed receptor density were detected; this shows the potential usefulness of the substance for diagnostic purposes, e.g., the differential diagnosis of various forms of epilepsy.

Effects of an intrathecally administered benzodiazepine receptor agonist, antagonist and inverse agonist on morphine-induced inhibition of a spinal nociceptive reflex.

1. The effects of an intrathecally administered benzodiazepine receptor (BZR) agonist (midazolam, up to 50 micrograms), antagonist (flumazenil, Ro 15-1788, 5 micrograms) and inverse agonist (Ro 19-4603, 15 micrograms) on nociception and on morphine-induced antinociception were studied in rats. 2. By themselves, none of these compounds significantly altered pain threshold. 3. The BZR agonist midazolam enhanced the morphine-induced antinociceptive effect whereas the antagonist flumazenil did not alter it. In contrast, the BZR inverse agonist Ro 19-4603 decreased the morphine-induced antinociceptive effect. 4. Naloxone (1 mg kg-1 i.p.) completely reversed all these effects. 5. These results demonstrate that BZR agonists and inverse agonists are able to affect, by allosteric up- or down-modulation of gamma-aminobutyric acidA (GABAA)-receptors, the transmission of nociceptive information at the spinal cord level, when this transmission is depressed by mu-opioid receptor activation.

Convulsions induced by centrally administered NMDA in mice: effects of NMDA antagonists, benzodiazepines, minor tranquilizers and anticonvulsants.

1. Convulsions were induced reproducibly by intracerebroventricular injection of N-methyl-D-aspartic acid (NMDA) to conscious mice. 2. Competitive (carboxypiperazine-propylphosphonic acid, CPP; 2-amino-7-phosphonoheptanoic acid, AP7) and non-competitive (MK801; phencyclidine, PCP; thienylcyclohexylpiperidine, TCP; dextrorphan; dextromethorphan) NMDA antagonists prevented NMDA-induced convulsions. 3. Benzodiazepine receptor agonists and partial agonists (triazolam, diazepam, clonazepam, Ro 16-6028), classical anticonvulsants (diphenylhydantoin, phenobarbitone, sodium valproate) and meprobamate were also found to prevent NMDA-induced convulsions. 4. Flumazenil (a benzodiazepine receptor antagonist) and the GABA agonists THIP and muscimol (up to subtoxic doses) were without effect. 5. Flumazenil reversed the anticonvulsant action of diazepam, but not that of MK801. 6. Results obtained in this model differ somewhat from those described in a seizure model with systemic administration of NMDA. An explanation for this discrepancy is offered. 7. This model is a simple test for assessing the in vivo activity of NMDA antagonists and also expands the battery of chemically-induced seizure models for characterizing anticonvulsants not acting at NMDA receptors.

Analysis of the serotonin transporter gene linked polymorphism (5-HTTLPR) in anorexia nervosa.

Previous studies have demonstrated aberrant expression of serotonin in individuals with an eating disorder. Given this the serotonin transporter gene (5-HTT) is a strong candidate to contribute to the genetic component of the aetiology of eating disorders. To determine the role of this particular gene in the susceptibility to anorexia nervosa (AN) we have examined a tandemly repeated sequence close to the promotor region of the 5-HTT gene, which is represented by a long (L) and short (S) variant. Previous studies have shown that the transcriptional activity of the 5-HTT gene differs significantly between these two alleles. A group of 138 Diagnostic and Statistical Manual of Mental Disorders (DSM-IV) criteria AN patients and 90 controls were genotyped at the 5-HTT gene linked polymorphism (5-HTTLPR). Statistical analysis showed no significant difference in allele or genotype frequencies between the two groups. These data suggest that there is no association between 5-HTTLPR genotype and susceptibility to AN, in our population. Am. J. Med. Genet. (Neuropsychiatr. Genet.) 96:53-55, 2000.

Benzodiazepine antagonist Ro 15-1788: neurological and behavioral effects.

In neurological and behavioral studies in mice, rats, dogs and squirrel monkeys, the imidazobenzodiazepinone Ro 15-1788 acted as a potent benzodiazepine antagonist. The antagonistic activity was both preventive and curative and seen at doses at which no intrinsic effects were detected. It was highly selective in that it acted against CNS effects induced by benzodiazepines but not against those produced by other depressants, such as phenobarbitone, meprobamate, ethanol, and valproate. The onset of action was rapid even after oral administration. Depending on the animal species studied, the antagonistic effects lasted from a few hours to 1 day. The acute and subacute toxicity of Ro 15-1788 was found to be very low. Benzodiazepine-like effects were not seen.

Endogenous oxidative stress induces distinct redox forms of tumor necrosis factor receptor-1 in melanoma cells.

Receptors of the TNFR superfamily possess abundant thiols in their extracellular domains, which makes them susceptible to redox modulation by prooxidant agents and processes. Previous studies from our laboratory have documented that membrane gamma-glutamyltransferase (GGT) activity can originate reactive oxygen species in the extracellular milieu, during the GGT-mediated metabolism of extracellular glutathione. The present study was aimed thus to verify a possible redox-modulating effect of GGT activity on TNFR1 receptors. The thiol-specific probe maleimide-polyethylene glycol was used to selectively label the reduced thiol groups in proteins of cell lysates; fractions corresponding to TNFR1 were then identified by immunoblot. In human melanoma Me665/2 cells, expressing varying GGT levels, at least five distinct forms of TNFR1 have been thus identified. The more oxidized forms appear to be prevalent in the 2/60 clone, expressing higher GGT levels, as compared to clone 2/21. Stimulation of GGT activity in the latter induced an increase of the oxidized TNFR1 forms. It is conceivable that different redox states of TNFR1 may correspond to different binding affinity and/or changes in the transducing function of the receptor. As GGT is frequently expressed by malignant tumors, the described phenomena might concur to alter the sensitivity of cancer cells to agents targeted on activation of TNF-alpha-dependent signaling pathways.

Disappearance of supra-ependymal 5-HT axons in the rat forebrain after electrolytic and 5,6-DHT-induced lesions of the medial forebrain bundle.

The present study was intended to demonstrate the origin of supra-ependymal 5-hydroxytryptamine axons in the rat forebrain. Electrolytic lesions and injections of 5,6-dihydroxytryptamine (10 mug in 4 mul) were carried out unilaterally in and close to the medial forebrain bundle in the posterior hypothalamus of rats. Ten to 14 days later, terminal axons and formaldehyde-induced indolealkylamine fluorescence had virtually disappeared supra-ependymally in the lateral ventricles and interventricular foramina ipsilateral to the lesion if the indolealkylamine axons passing through the medial part of the medial forebrain bundle had been destroyed. No changes were observed, electron microscopically or fluorescence histochemically, in ventricles contralateral to the lesion. It is concluded that the supra-ependymal serotonergic nerve terminals in the lateral ventricles and interventricular foramina originate, uncrossed, from non-terminal axons passing through the medial forebrain bundle in the posterior hypothalamus.

Isoniazid-induced convulsions in rats: effects of Ro 15-1788 and beta-CCE.

Dose-response curves for the convulsant effect of isoniazid were determined in male rats. The specific benzodiazepine antagonist Ro 15-1788, at doses from 20 to 100 mg/kg i.v. or at 300 mg/kg p.o., failed to produce any relevant shift of the isoniazid dose-response curves to the left (proconvulsant) or to the right (anticonvulsant). In contrast, the benzodiazepine receptor ligand ethyl-beta-carboline-3-carboxylate (beta-CCE) 10 mg/kg i.v. produced a clear-cut shift to the left of the isoniazid dose-response curve. Ro 15-1788 was inactive up to high p.o. or i.v. doses, versus convulsions induced by a low supraliminal dose of isoniazid, whereas beta-CCE had a significant proconvulsant effect beginning at 5 mg/kg i.v. This effect of beta-CCE was dose dependently antagonized by Ro 15-1788. In conclusion, in the present experimental conditions Ro 15-1788 showed no inverse agonistic properties at benzodiazepine receptors, in contrast to beta-CCE.

Physical dependence induced in DBA/2J mice by benzodiazepine receptor full agonists, but not by the partial agonist Ro 16-6028.

Continuous administration of triazolam, alprazolam or diazepam for a 7-day period by means of minipumps or chronic (17 days) p.o. treatment with alprazolam induced clear physical dependence in DBA/2J mice as assessed by precipitation of a withdrawal syndrome with an i.v. injection of the benzodiazepine receptor partial inverse agonist Ro 15-3505. In contrast, no precipitated withdrawal signs were observed following chronic exposure to high doses of the benzodiazepine receptor partial agonist Ro 16-6028. The use of minipumps and precipitation with a benzodiazepine receptor antagonist permits a simple and rapid evaluation of the physical dependence liability of potent compounds acting at the benzodiazepine receptor. Furthermore, these results support the hypothesis that benzodiazepine receptor partial agonists are less likely to induce physical dependence than full agonists.

The effect of diphenylhydantoin, diazepam and clonazepam on the activity of Purkinje cells in the rat cerebellum.

Spike discharges of single cerebellar Purkinje cells were recorded continuously with extracellular microelectrodes in unanesthetized curarized rats. The intravenous injection of diphenylhydantoin in doses between 10 and 100 mg kg-1 did not substantially alter the activity of Purkinje cells within 2--3 h. The two benzodiazepines, diazepam and clonazepam, already in low i.v. doses (0.03--0.1 mg kg-1) consistently and reversibly depressed the firing rate. Our results do not support the previously advanced hypothesis that these drugs reduce epileptiform activities by increasing the output from the cerebellar cortex. They rather point to the possibility that a reduced firing rate of cerebellar Purkinje cells mediates at least in part ataxia and muscular hypotonia observed after the drugs.

Improved rapid method to check electrode localizations in the brain.

A modification of known rapid methods to check electrode localization in the brain is described: the essential point consists in immersing the brain, after fixation in 4 percent Formalin, in pure heptane at -30 degrees C for 5 hr. Unstained sections, cut with the freezing microtome, are placed uncovered on glass slides and photographed (in an enlarger, as if they were negatives) either wet, to obtain a contrast resembling Weil staining, or dry, to obtain a picture corresponding to Nissl staining.

Fine mapping of the human 5-HTR2a gene to chromosome 13q14 and identification of two highly polymorphic linked markers suitable for association studies in psychiatric disorders.

The serotonergic system is known to play an important role in a number of psychiatric disorders. Indeed, treatments involving agents that have their pharmacological activities within this system are the mainstay of treatment for disorders such as schizophrenia. It is now widely accepted that many common psychiatric disorders have a familial or genetic component and as a result of this there has been an upsurge in interest in the 5-hydroxytryptamine (5-HT) pathways. A number of groups have attempted to establish whether polymorphism in particular proteins of the serotonergic system may form part of the genetic component of psychiatric disorders, including schizophrenia and anorexia nervosa. However, the data from these studies are conflicting and the problem is compounded by the lack of known polymorphic genetic markers mapping in close proximity to genes encoding proteins envolved directly or indirectly in 5-HT neurotransmission. In the current study, we have fine mapped the gene for 5-HTR2a by radiation hybrid mapping, and we report two new, highly linked, polymorphic markers that are suitable for linkage and association studies.

Sarmazenil-precipitated withdrawal: a reliable method for assessing dependence liability of benzodiazepine receptor ligands.

The benzodiazepine receptor partial inverse agonist sarmazenil exhibits in vivo proconvulsive, but not convulsant, effects in different paradigms in rodents. Intravenous sarmazenil challenge given at several fixed intervals following the termination of repeated treatment with a markedly sedative dose of diazepam in squirrel monkeys was effective in precipitating withdrawal signs, but had no comparable effects in vehicle-treated controls. The precipitated withdrawal reaction was not only robust, but it was consistently observed in all of the diazepam-treated monkeys. Thus, the use of sarmazenil challenge in the precipitated withdrawal paradigm provides a reliable method for assessing the development of physical dependence during repeated treatment with benzodiazepine receptor agonists.