Evaluation of the Bactericidal Activity of Plazomicin and Comparators against Multidrug-Resistant Enterobacteriaceae.

The next-generation aminoglycoside plazomicin, in development for infections due to multidrug-resistant (MDR) Enterobacteriaceae, was evaluated alongside comparators for bactericidal activity in minimum bactericidal concentration (MBC) and time-kill (TK) assays against MDR Enterobacteriaceae isolates with characterized aminoglycoside and ß-lactam resistance mechanisms. Overall, plazomicin and colistin were the most potent, with plazomicin demonstrating an MBC50/90 of 0.5/4 µg/ml and sustained 3-log10 kill against MDR Escherichia coli, Klebsiella pneumoniae, and Enterobacter spp.

Effect of Origanum vulgare essential oil on biofilm formation and motility capacity of Pseudomonas fluorescens strains isolated from discoloured Mozzarella cheese.

AIMS: The study was focused on Pseudomonas fluorescens strains isolated from Mozzarella cheese, with the aim of evaluating the effects of Origanum vulgare L. essential oil (OEO) on the biofilm formation and eradication, as well as on the motility and blue pigment production at 10°C. METHODS AND RESULTS: Microdilution method was used to determine the minimum inhibitory and bactericidal concentration of the OEO, which ranged between 10 and 40 µl ml-1 . In vitro studies demonstrated that a sublethal concentration of OEO influenced not only P. fluorescens growth and motility but also the capability to form biofilm and, in a lower degree, the biofilm eradication at 10°C. Analysis by confocal microscopy revealed a dramatic reduction in biofilm formation and thickness, with scattered damage or death of cells, stained by propidium iodide. In addition, a concentration of 5 µl ml-1 of OEO affected the motility of the cells and, in particular, their ability to swim. However, the essential oil did not inhibit the blue pigment production by any of the tested strains. CONCLUSIONS: The present findings suggest that oregano essential oil inhibits the biofilm formation of P. fluorescens strains and alters their motility. Moreover, in the preformed biofilm, OEO contributes to the detachment of the cells, deteriorating the architecture of the biofilm and reducing its thickness. SIGNIFICANCE AND IMPACT OF THE STUDY: The O. vulgare L. essential oil was revealed as a promising agent against biofilm formation and for its detaching; these results suggest that oregano EO could be used in the dairy food industry to control biofilm formation, as an alternative, or in combination with conventional sanitizers.

Diversity of food-borne Bacillus volatile compounds and influence on fungal growth.

AIMS: To evaluate the antifungal activity of the volatile organic compounds (VOCs) produced by 75 different food-borne Bacillus species against Aspergillus niger, Aspergillus flavus, Aspergillus parasiticus, Aspergillus clavatus, Fusarium oxysporum f. sp. lactucae and Moniliophthora perniciosa and to determine the VOCs responsible for the inhibition. METHODS AND RESULTS: Bacillus strains inhibited fungal growth, although with different inhibition grades, with Bacillus subtilis, Bacillus amyloliquefaciens and Bacillus cereus strains as the best antifungal VOCs producers. While M. perniciosa DM4B and F. oxysporum f.sp. lactucae MA28 were the most sensitive fungi, A. parasiticus MG51 showed the greatest resistance to Bacillus VOCs exposure. Thirty-seven compounds were detected by SPME-GC-MS analysis, although similar patterns in volatile compounds were evidenced within the species, interspecific VOCs differences determined different effects on fungal growth. Multiple partial least regression (MPLRS) and antifungal activity of the individual VOCs revealed that only propanone, 1-butanol, 3-methyl-1-butanol, acetic acid, 2-methylpropanoic acid, carbon disulphide, 3-methylbutanoic acid and ethyl acetate were responsible for mycelia inhibition of M. perniciosa DM4B and F. oxysporum f.sp. lactucae MA28. CONCLUSIONS: The antagonistic activity of the Bacillus VOCs was demonstrated, although it cannot easily be explained through the action of a single molecule, thus a holistic approach could be more appropriate to estimate the fungal growth inhibition. SIGNIFICANCE AND IMPACT OF THE STUDY: VOCs produced by Bacillus from cooked food can be considered as promising antifungal compounds useful in the control of fungal plant pathogens. This study investigates for the first time the correlation between mycelia inhibition of M. perniciosa and F. oxysporum f. sp. lactucae and the VOCs emitted by the Bacillus species.

Evaluation of omadacycline against intracellular Mycobacterium abscessus in an infection model in human macrophages.

Background: Omadacycline is an aminomethylcycline antibiotic in the tetracycline class that was approved by the US FDA in 2018 for the treatment of community-acquired bacterial pneumonia and acute bacterial skin and skin structure infections. It is available in both IV and oral formulations. Omadacycline has broad-spectrum in vitro activity and clinical efficacy against infections caused by Gram-positive and Gram-negative pathogens. Omadacycline is being evaluated in a 3 month placebo-controlled Phase 2 clinical trial of oral omadacycline versus placebo in adults with non-tuberculous mycobacteria (NTM) pulmonary disease caused by Mycobacterium abscessus (NCT04922554). Objectives: To determine if omadacycline has intracellular antimicrobial activity against NTM, bacteria that can cause chronic lung disease, in an ex vivo model of intracellular infection. Methods: Two strains of M. abscessus were used to infect THP-1 macrophages. Intracellular M. abscessus was then challenged with omadacycline and control antibiotics at multiples of the MIC over time to evaluate intracellular killing. Results: At 16 ×  the MIC at 72 h, omadacycline treatment of intracellular NTM yielded a log10 reduction in cfu of 1.1 (91.74% reduction in cfu) and 1.6 (97.65% reduction in cfu) consistent with killing observed with tigecycline, whereas amikacin and clarithromycin at 16 ×  the MIC did not show any reduction in cfu against the intracellular M. abscessus. Conclusions: Omadacycline displayed intracellular activity against M. abscessus within macrophages. The activity was similar to that of tigecycline; as expected, intracellular killing was not observed with clarithromycin and amikacin.

Familial dilated cardiomyopathy. Clinical and genetic characteristics.

Familial dilated cardiomyopathy (F-DCM) describes a clinically and genetically heterogeneous group of diseases, mostly inherited as autosomal dominant traits, having idiopathic left ventricular dilatation and dysfunction as a common phenotype. The age of onset, rate of progression, disease complications, as well as overall prognosis and outcome vary both amongst and within families. Clinical traits, both cardiac and extracardiac, may recur in association with the DCM phenotype. The former include conduction defects, structural abnormalities such as left ventricular noncompaction, of right ventricular involvement, and recurrence of atrial or ventricular arrhythmias; the latter commonly affect the musculoskeletal (myopathies/dystrophies, both clinically overt and subclinical), ocular, auditory, nervous, and integument systems. These traits may help guide genetic testing. In parallel to the clinical heterogeneity, F-DCM also shows genetic heterogeneity: more than 40 genes have been causally linked to F-DCM, with mutations recurring more commonly in a few known genes, and less frequently in rare, less commonly known genes. Based on the known prevalence of mutations in disease genes, more than 50% of F-DCM cases can be regarded as still genetically orphan, implying that further disease genes have to be discovered. Family screening and genetic testing are now established as the gold standard for diagnosis, care, and prevention in F-DCM. Diagnostic tests are performed using Sanger-based sequencing. Furthermore, new biotechnology tools, based on next-generation sequencing, are now being implemented in the research setting and will dramatically modify the future of the nosology of F-DCM.

[Dose and effect biomarkers in the diagnosis of asbestos-related diseases and health surveillance of exposed workers]. / Gli indicatori biologici di dose ed effetto nella diagnostica delle patologie asbesto-correlate e nella sorveglianza sanitaria degli ex-esposti.

The aim of the study was to assess the reliability of the indicators of dose and effect in the health monitoring of asbestos exposed workers. In 49 cases out of 158 studied workers (31%) asbestos-related diseases were diagnosed following ATS criteria (2004). Using nonparametric statistical methods (permutation tests) 6 variables were analyzed with respect to asbestos-related diseases and working sectors, demonstrating a difference in the concentration of amphiboles (p < 0.01), greater in patients with asbestosis and workers involved in asbestos removal from railway carriages. There was not a correlation between mesothelin and amphiboles, chrysotile and total fibers concentrations (Spearman test).

Neuropsychological manifestations of long COVID in hospitalized and non-hospitalized Brazilian Patients.

BACKGROUND: There has been a significant increase in number of patients seeking neuropsychological rehabilitation months after the acute phase of COVID-19 infection. OBJECTIVE: Identify the cognitive and psychiatric disorders in patients with long COVID or Post-Acute Sequelae of COVID (PASC) and explore the association between disease severity during the acute phase and persistent neuropsychological manifestations. METHODS: 614 adults were assessed an average of eight months post-infection. Participants were, on average, 47.6 y.o., who sought rehabilitation for neuropsychological problems. Patients were evaluated using the Barrow Neurological Institute Screen for Higher Cerebral Functions (BNIS), Phonemic Verbal Fluency and Clock Drawing tests (NEUPSILIN) for executive functions, and the Hospital Anxiety and Depression Scale (HADS). RESULTS: The BNIS score was significantly below reference values in all subscales, especially affect and memory. Verbal Fluency and Clock Drawing subtest results were also lower. Patients with PASC tested high for anxiety/depression, but there was no statistically significant relationship between HADS and BNIS scores. Neuropsychological evaluations showed no differences in cognitive or psychiatric profiles between hospitalized and non-hospitalized patients. CONCLUSIONS: Neuropsychological results suggest executive function problems and high incidence of anxiety/depression, irrespective of acute-phase severity, underscoring a need for neurorehabilitation programs while providing data for public policy initiatives.

Electronic paramagnetic resonance investigation of the activity of Origanum vulgare L. essential oil on the Listeria monocytogenes membrane.

AIMS: To evaluate the effect of oregano essential oil on Listeria monocytogenes cytoplasmic membrane. METHODS AND RESULTS: Nitroxide free-radical Electron Paramagnetic Resonance was applied on L. monocytogenes after 30 min exposure to oregano essential oil concentrations ranging from 0 to 1.25%. The impact of essential oil on the number of viable cells was evaluated by plate count. Growth dynamics of survivors in BHI and TSB were evaluated by turbidometry. After exposure to essential oil concentrations up to 0.50%, the membrane fluidity was changed and its order increased. When L. monocytogenes was exposed to higher concentrations, membrane order parameters slightly returned to the values of untreated cells. However, when the cells were exposed to EO in the presence of sodium azide, which impairs energy metabolism, the membrane fluidity was progressively enhanced, even at the lowest EO concentration (0.25%). Microbiological analyses confirmed a progressive reduction of viable count, at increasing essential oil concentrations. Both in BHI and TSB, the Lag phase length increased in treated cells with respect to controls, suggesting a cell damage recovery. CONCLUSIONS: The combined approach including microbiological and EPR analyses provided relevant information on membrane modification and cell response to essential oils. SIGNIFICANCE AND IMPACT OF THE STUDY: EPR approach was demonstrated to be an effective and helpful tool to comprehend the modifications exerted by essential oil on the bacterial membrane.

Activity of plazomicin in combination with other antibiotics against multidrug-resistant Enterobacteriaceae.

Plazomicin is a next-generation aminoglycoside that was approved by the US FDA in June 2018 for the treatment of complicated urinary tract infections (cUTIs), including pyelonephritis due to Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae and Proteus mirabilis. Plazomicin is active against multi-drug resistant (MDR) Enterobacteriaceae, where combination therapy is often used to treat infections caused by these pathogens. To determine synergy with other antibiotics, plazomicin was combined with antibiotics in checkerboard assays against MDR Enterobacteriaceae, including isolates with resistance to aminoglycosides and ß-lactams; 10 Escherichia coli isolates, 8 Klebsiella spp. isolates, 10 Enterobacter spp. isolates, and 2 Citrobacter freundii isolates were evaluated. Plazomicin had potent activity against MDR Enterobacteriaceae, including aminoglycoside-resistant strains, with MIC ranges of 0.5 - 2 µg/mL against E. coli isolates, 0.12 - 8 µg/mL against Klebsiella spp. isolates, 0.25 - 2 µg/mL against Enterobacter spp. isolates, and 0.06 - 0.25 µg/mL against C. freundii isolates. Synergy between plazomicin and piperacillin/tazobactam or ceftazidime was observed by checkerboard studies and confirmed by time-kill assays. No combination showed antagonism. These studies indicate that plazomicin has potential as a monotherapy and as combination therapy for treating serious Gram-negative infections caused by MDR Enterobacteriaceae.

Antimicrobial activity of Austroeupatorium inulaefolium (H.B.K.) against intracellular and extracellular organisms.

The antimicrobial activity of Austroeupatorium inulaefolium (H.B.K.) essential oil was studied in different pathogens species and its cytotoxicity activity was determinated on different cellular lines. Despite the good antibacterial activity of A. inulaefolium, it has been cytotoxic at low concentrations. Consequently it might be interesting to determine the antimicrobial activity and cytotoxicity of the major compounds of this essential oil.

Increased expression of peripheral benzodiazepine receptors on leukocytes in silent myocardial ischemia.

OBJECTIVES: The purpose of this study was to evaluate benzodiazepine receptor expression on leukocytes from patients with symptomatic or silent myocardial ischemia. BACKGROUND: Silent myocardial ischemia is frequently observed in patients with coronary artery disease. Pain can be effectively controlled by various endogenous mechanisms. Benzodiazepines and their receptors play key roles in pain, in interactions with peptide opioids, in inflammation and in the response to stress. METHODS: The study group consisted of 57 patients with reproducible exercise-induced myocardial ischemia. The presence of a constant behavior in the anginal pain perception during both exercise-induced ischemia and daily life was the most important inclusion criterion. Venous blood samples were taken from all patients to evaluate the expression of peripheral benzodiazepine receptors by flow cytometry. The study cohort was classified into two groups: 24 patients who had anginal pain both at home and during the exercise stress test and 33 patients who were asymptomatic during both daily life and exercise-induced ischemia. RESULTS: Flow cytometry analysis showed increased expression of peripheral benzodiazepine receptors on all types of leukocytes in the asymptomatic patients. The difference was statistically significant for lymphocytes (p < 0.005), monocytes (p < 0.001) and granulocytes (p < 0.001). CONCLUSIONS: These data show that expression of peripheral benzodiazepine receptors was higher in patients with silent myocardial ischemia than in symptomatic patients.

In vitro binding of ciliary neurotrophic factor to its receptors: evidence for the formation of an IL-6-type hexameric complex.

Ciliary neurotrophic factor (CNTF) is a cytokine sharing structural and functional similarities with interleukin-6 (IL-6) and other helical cytokines that utilize the common signalling chain gp130. While IL-6 induces gp130 dimerization, CNTF, after the initial interaction with the specific, non-signalling receptor subunit, CNTFR, induces the formation of gp130/LIF-receptor heterodimers. Through immunoprecipitation experiments with tagged soluble receptor molecules, we recently demonstrated that IL-6 drives the formation of a hexameric receptor complex with a defined topology and composed of two IL-6, two IL-6R alpha and two gp130 molecules. Here, we apply the same strategy to study the assembly in vitro of the CNTF receptor complex. We present evidence that both the cytokine and the specific binding chain undergo dimerization in the presence of gp130. Furthermore, although gp130 and LIFR are able to bind independently to the CNTF/CNTFR sub-complex, they never form homodimers but only heterodimers. We propose that CNTF assembles a hexameric receptor complex composed of two CNTF, two CNTFR, one gp130 and one LIFR molecule, and present a model of the reciprocal interaction of these molecules based on similarities with the IL-6 hexameric complex.

Prevention and treatment of nephrolithiasis: a review on the role of spa therapy.

The prevalence and incidence of nephrolithiasis is reported to be increasing across the world. It is a disease of increased urinary concentration of stone-forming salts. The physicochemical mechanism of stone formation includes precipitation, homogenous/heterogeneous nucleation, growth, aggregation and concretion of various modulators in urine. Necessary condition to develop stones is urinary supersaturation, due to reduced urinary volume or to an excesses solutes. Fluid intake is the main determinant of urine volume. Urine dilution can significantly decrease both the crystallization rate of the urinary salts and the aggregation of the crystals. A correct fluid intake can act on different effects: urinary tract washing, urinary volume increasing and dilution of solutes. In addition mineral waters have other particular features: greater diuretic effect, more important urinary dilution with solutes and microbial concentration reduction, urinary pH changes, superior washout effect due to mechanical effects and ureteral contractions. Adequate water intake is the most important conservative strategy in urolithiasis prevention; particularly hydropinotherapy with oligomineral water should be considered as an important instrument to prevent stones in subjects predisposed to the disease (family members of people suffering from kidney stones), to reduce relapses, and can help to eliminate residual fragments also after extracorporeal shock wave lithotripsy. It is recommended a management with increased mineral water intake to promote urine volume of at least 2.5L each day to prevent stone formation. Obviously water intake shall be varied in relation to the presence of contraindications or any diseases.

[Ethics in clinical practice and in health care]. / L'etica nella pratica clinica e nell'assistenza sanitaria.

The clinical ethics is the identification, analysis and solution of moral problems that can arise during the care of a patient. Given that when dealing with ethical issues in health care some risks will be encountered (talking about ethics in general, or as a problem overlapped with others in this area, or by delegation to legislative determinations) in the text certain important aspects of the topic are examined. First of all ethics as human quality of the relationship between people for the common good, especially in health services where there are serious problems like the life and the health. It is also necessary a "humanizing relationship" between those who work in these services in order to achieve quality and efficiency in this business. It is important a proper training of health professionals, especially doctors, so that they can identify the real needs and means of intervention. It is also important that scientific research must respect fundamental ethical assumptions. In conclusion, ethics in health care is not a simple matter of "cookbook" rules, but involves the responsibility and consciousness of individual operators.

Association between 5-HT(2A) receptor polymorphism and psychotic symptoms in Alzheimer's disease.

BACKGROUND: The aim of this study is to analyze the segregation of the 102T/C polymorphism in the serotonin 2A receptor gene in patients affected by sporadic and familial Alzheimer's disease (FAD) with and without psychotic symptoms. METHODS: The polymorphism was analyzed in 275 subjects. A semistructured interview was used to obtain information about delusions, hallucinations, and other specific behavioral signs occurring during the disease. RESULTS: Fifty-two percent of AD patients with psychotic symptoms were homozygous for the C102 allele, as compared with 6.9% of AD patients without psychosis. Similarly, the C102/C102 genotype was significantly more frequent in FAD patients with psychosis than in FAD patients without (46.5% vs. 7.8%). CONCLUSIONS: Our data strongly confirm and extend to FAD previous studies suggesting that the genetic variation at this locus is associated with prominent psychotic features in AD and that the 102C allele could play an important role in late-onset AD.

Genetic and clinical analysis of spinocerebellar ataxia type 8 repeat expansion in Italy.

BACKGROUND: The spinocerebellar ataxias (SCAs) are clinically heterogeneous disorders caused by triplet repeat expansions in the sequence of specific disease genes. Spinocerebellar ataxia type 8 (SCA8), originally described in a family characterized by pure cerebellar ataxia with slow disease progression, presents with expansion of combined CTA/CTG repeats. OBJECTIVE: To perform SCA8 repeat expansion analysis in a heterogeneous group of ataxic patients, to determine the prevalence of this mutation in our patients and establish the frequency of expanded CTA/CTG repeats in a large group of control subjects. PATIENTS: One hundred sixty-seven patients affected by sporadic, autosomal dominant and recessive hereditary ataxia were clinically examined and analyzed for SCA8 expansion. We further studied 161 control subjects and 125 patients with psychiatric disorders. RESULTS: We found abnormally expanded CTA/CTG repeats in 5 ataxic patients, 3 of them characterized by pure cerebellar ataxia. One patient had vitamin E deficiency and 1 patient with a sporadic case was affected by gluten ataxia. No evidence of expanded alleles was found in healthy control subjects and in patients with psychiatric disorders. CONCLUSIONS: Our data support the evidence that CTG expansions may be linked to SCA8, since the pathogenic expansions have been found only among patients with genetically unidentified forms of hereditary and sporadic ataxia. Patients carrying expanded alleles present peculiar phenotypic features, thus suggesting that unknown additional factors could probably predispose to the disease.

Measurement of bone mineral density at the spine and proximal femur by volumetric quantitative computed tomography and dual-energy X-ray absorptiometry in elderly women with and without vertebral fractures.

The goal of this study was to determine the effect of vertebral fracture status on trabecular bone mineral density (BMD) measurements obtained in the proximal femur and spine by helical volumetric quantitative computed tomography (vQCT). The study population consisted of 71 Italian women (average age 73 +/- 6) years. This group included 26 subjects with radiographically confirmed atraumatic vertebral fractures and 45 controls. The subjects received helical CT scans of the L1 and L2 vertebral bodies and the hip. The three-dimensional CT images were processed using specialized image analysis algorithms to extract measurements of trabecular, cortical, and integral BMD in the spine and hip. To compare the vQCT results with the most widely used clinical BMD measurement, dual X-ray absorptiometry (DXA) scans of the anteroposterior (AP) spine and proximal femur were also obtained. The difference between the subjects with vertebral fractures and the age-matched controls was computed for each BMD measure. All BMD measurements showed statistically significant differences, which ranged from 7% to 22% between subjects with fractures and controls. Although, given our small sample size, we could not detect statistically significant differences in discriminatory power between BMD techniques, integral BMD of the spine measured by vQCT and DXA tended to show stronger associations with fracture status (0.001 < p < 0.004). Measurements by QCT and DXA at the hip were also associated with vertebral fracture status, although the association of DXA BMD with fracture status was explained largely by differences in body weight between subjects with vertebral fractures and controls.

Planum temporale asymmetry and thought disorder in schizophrenia.

Magnetic resonance imaging (MRI) was used to investigate the size and the asymmetry of the planum temporale (PT) in 22 schizophrenic patients and 23 strictly matched healthy volunteers. The degree of thought disorder was related to the reduction of the physiological PT asymmetry. When thought disordered patients were contrasted with non-thought disordered patients and healthy controls for a measure of PT laterality, those with thought disorder showed a statistically significant loss of PT laterality. This finding suggests that thought disordered schizophrenics may be characterized by an abnormal development of cerebral lateralization in a region crucial for language processing.

Cortical and subcortical computed tomographic study in schizophrenia.

Thirty-six chronic schizophrenics and 24 controls, all males, were evaluated by computed tomography (CT) scans. The lateral, third and fourth ventricles, the Sylvian fissure and the largest sulcus from each lobe were measured. In the schizophrenic patients there was a significant increase in third ventricle, ventricular brain ratio (VBR) and cortical measures. The possible implications of these findings in the aetiopathology of schizophrenia are discussed.

Analysis of apolipoprotein E, alpha1-antichymotrypsin and presenilin-1 genes polymorphisms in dementia caused by normal pressure hydrocephalus in man.

Normal pressure hydrocephalus (NPH) is characterized by dementia, gait disorders and urinary incontinence. Apolipoprotein E (ApoE) epsilon4 allele has been associated with severity of dementia in Alzheimer's disease (AD) and in other forms of dementia. Moreover, homozygosity of the A allele of the alpha1-antichymotrypsin (ACT) gene and of allele 1 of the presenilin-1 (PS-1) gene was associated with an increased risk for late onset AD. We analyzed the distribution of ApoE, ACT and PS-1 genotypes and the corresponding allele frequencies in 13 NPH patients. No differences were found in ACT and PS-1 polymorphism distributions in the patients studied with respect to the control group. An increased ApoE epsilon4 allele frequency was observed in NPH patients with respect to controls, thus suggesting that epsilon4 allele may also be involved in the pathogenesis of the disease.