Atmosferic pressure non-thermal plasma: Preliminary investigation

Antibacterial activity of atmosferic pressure non-thermal plasma (APNTP) was assessed for bacterial, yeast and mold strains. This investigation is to be considered preliminary: a second step is envisaged in which the efficacy of the technique and the device will be assessed directly on food of animal and plant origin. The strains (ATCC or wild type) of Listeria innocua, Escherichia coli, Salmonella thyphimurium, Pseudomonas aeruginosa, Staphylococcus aureus, Enterococcus faecalis, Proteus mirabilis (bacteria); Alternaria alternata, Aspergillus flavus, Cladosporium herbarum, Fusarium graminearum, Geotrichum candidum, Penicillium roqueforti, Rhizopus nigricans (moulds); Candida parapsilosis and Candida albicans (yeasts) were subjected to plasma plume generated by the action of electric fields with a gas mixture (oxygen and helium) delivered for 5 min at a distance of 2 cm. Types of experiments were listed as following: microorganism at concentration 1×10^8 and 1×104 cfu on PCA (Plate Count Agar); Listeria innocua and Salmonella thiphymurium at concentration 1×10^4 cfu on semi-synthetic and synthetic medium; mycetes (moulds and yeasts) at concentration 1×10^8 and 1×10^4 cfu on SDA (Sabouraud Dextrose Agar). The results obtained on the bacteria subjected to atmospheric cold plasma were evident on all the strains tested except for Proteus mirabilis (1×10^8 cfu), most evident at a concentration of 1×10^4 cfu, not only on culture media PCA but also on semi-synthetic medium and jelly meat-PCA medium. In spite of bacterial results, treatment with plasma plume did not decrease or inhibit of fungal growth. That means plasma plume was neither fungicidal nor fungistatic activities

The Relationship between Heart Rate Variability and Adiposity Differs for Central and Overall Adiposity

While frank obesity is associated with reduced HRV, indicative of poorer autonomic nervous system (ANS) function, the association between body mass index (BMI) and HRV is less clear. We hypothesized that effects of adiposity on ANS are mostly mediated by visceral fat and less by subcutaneous fat; therefore, centrally distributed adipose tissue, that is, waist circumference (WC), should be more strongly associated with HRV than overall adiposity (BMI). To examine this hypothesis, we used data collected in a subset of the Baltimore Longitudinal Study of Aging to compare strength of association between HRV and WC to that of HRV and BMI. Time domain HRV variables SDNN (standard deviation of successive differences in normal-to-normal (N-N) intervals) and RMSSD (root mean square of successive differences in N-N intervals) were calculated from 24-hour Holter recordings in 159 participants (29–96 years). Increasing WC was associated with decreasing SDNN and RMSSD in younger but not older participants (P value for WC-by-age interaction = 0.003). BMI was not associated with either SDNN or RMSSD at any age. In conclusion, central adiposity may contribute to sympathetic and parasympathetic ANS declines early in life

White Blood Cell Count and Mortality in the Baltimore Longitudinal Study of Aging

ObjectivesWe investigated the secular trend in white blood cell (WBC) count and the relationship between WBC count and mortality between 1958 and 2002.BackgroundThe WBC count is a clinical marker of inflammation and a strong predictor of mortality. Limited data exist on the WBC count secular trend and the relationship between WBC and mortality.MethodsOne thousand eighty-three women and 1,720 men were evaluated longitudinally in the Baltimore Longitudinal Study of Aging. Blood samples and medical information were collected at the study entry and every 2 years during follow-up visits. The WBC count and all-cause, cardiovascular, and cancer mortality were assessed.ResultsA downward trend in WBC count was observed from 1958 to 2002. The secular downward trend was independent of age, gender, race, smoking, body mass index, and physical activity. The WBC count was nonlinearly associated with all-cause mortality and almost linearly associated with cardiovascular mortality. Participants with baseline WBC <3,500 cells/mm3and WBC >6,000 cells/mm3had higher mortality than those with 3,500 to 6,000 WBC/mm3. Within each WBC group, age-adjusted mortality rates declined in successive cohorts from the 1960s to the 1990s. Participants who died had higher WBC than those who survived, and the difference was statistically significant within 5 years before death.ConclusionsOur study provides evidence for a secular downward trend in WBC count over the period from 1958 to 2002. Higher WBC counts are associated with higher mortality in successive cohorts. We found no evidence that the decline of age-specific mortality rates that occurred from 1960 to 2000 was attributable to a secular downward trend in WBC

Rapamycin promotes differentiation increasing βIII-tubulin, NeuN, and NeuroD while suppressing nestin expression in glioblastoma cells

Glioblastoma cells feature mammalian target of rapamycin (mTOR) up-regulation which relates to a variety of effects such as: lower survival, higher infiltration, high stemness and radio- and chemo-resistance. Recently, it was demonstrated that mTOR may produce a gene shift leading to altered protein expression. Therefore, in the present study we administered different doses of the mTOR inhibitor rapamycin to explore whether the transcription of specific genes are modified. By using a variety of methods we demonstrate that rapamycin stimulates gene transcription related to neuronal differentiation while inhibiting stemness related genes such as nestin. In these experimental conditions, cell phenotype shifts towards a pyramidal neuron-like shape owing long branches. Rapamycin suppressed cell migration when exposed to fetal bovine serum (FBS) while increasing the cell adhesion protein phospho-FAK (pFAK). The present study improves our awareness of basic mechanisms which relate mTOR activity to the biology of glioblastoma cells. These findings apply to a variety of effects which can be induced by mTOR regulation in the brain. In fact, the ability to promote neuronal differentiation might be viewed as a novel therapeutic pathway to approach neuronal regeneration

Rapamycin dose-dependently promotes differentiation and cell death in an in vitro model of malignant gliomas

Glioblastoma (GB, grade IV astrocytoma) is the most common and lethal brain tumor characterized by increased proliferation and resistance to chemotherapy and radiotherapy. GB infiltrates the brain, always relapses and leads to death within 2 years from diagnosis. At cellular level, relapse and infiltration are correlated with the presence of GB precursors stem-like cells. At molecular level, relapse and infiltration are correlated with upregulation of the mammalian target of rapamycin, mTOR which constantly characterizes malignant gliomas. By definition mTOR is strongly inhibited by rapamycin and rapalogs. Several studies were carried out to evaluate the effects of rapamycin in experimental models of GB. However, it remains controversial whether rapamycin and rapalogs produce cell differentiation or cell death. Therefore, in the present study we used a wide range of doses of rapamycin to address the issue of differential effects obtained by rapamycin in various experimental setting. In particular, rapamycin was administered ranging from 1 nM up to 1microM to assess cell viability in GB cell lines (U87MG). All the experiments were carried out exposing the cells for 24 h to rapamycin. We found that rapamycin produced dose dependently cell death which was significant for doses starting at 100 nM and reaches a plateau at 1 microM (50% cell death). Interestingly, when using rapamycin at doses between 1 nM and 100 nM we described only slight cell death while the prominent effect induced by rapamycin consisted of cell differentiation. In detail, beta 3 tubulin increased, while nestin decreased under rapamycin exposure. These markers were related to phenotypic changes consisting of an increased number and length of cell processes along with the loss of fusiform cell shape. In the process of characterizing the effects of mTOR up-regulation and its pharmacological inhibition we also measured the accumulation of an mTOR-dependent protein such as alpha-synuclein. In fact, mTOR up-regulation is known to inhibit the autophagy pathway which removes alpha-synuclein. Consistently, in U87MG cells we found accumulation of alpha-synuclein which was removed dose dependently by rapamycin exposure. Our data show that inhibiting mTOR with rapamycin is a powerful tool to induce cell differentiation in U87MG cells, whereas cell death significantly occurs only at the highest doses

Sunny holidays before and after melanoma diagnosis are respectively associated with lower breslow thickness and lower relapse rates in Italy

Background: Previous studies have reported an association between sun exposure and improved cutaneous melanoma (CM) survival. We analysed the association of UV exposure with prognostic factors and outcome in a large melanoma cohort. Methods: A questionnaire was given to 289 (42%) CM patients at diagnosis (Group 1) and to 402 CM patients (58%) during follow-up (Group 2). Analyses were carried out to investigate the associations between sun exposure and melanoma prognostic factors and survival. Results: Holidays in the sun two years before CM diagnosis were significantly associated with lower Breslow thickness (p=0.003), after multiple adjustment. Number of weeks of sunny holidays was also significantly and inversely associated with thickness in a dose-dependent manner (p=0.007). However when stratifying by gender this association was found only among women (p=0.0004) the risk of CM recurrence in both sexes was significantly lower in patients (n=271) who had holidays in the sun after diagnosis, after multiple adjustment including education: HR=0.30 (95%CI:0.10-0.87; p=0.03) conclusions: Holidays in the sun were associated with thinner melanomas in women and reduced rates of relapse in both sexes. However, these results do not prove a direct causal effect of sun exposure on survival since other confounding factors, such as vitamin D serum levels and socio-economic status, may play a role. Other factors in sun seeking individuals may also possibly affect these results

Sequential immunotherapy and targeted therapy for metastatic BRAF V600 mutated melanoma: 4-year survival and biomarkers evaluation from the phase II SECOMBIT trial

No prospective data were available prior to 2021 to inform selection between combination BRAF and MEK inhibition versus dual blockade of programmed cell death protein-1 (PD-1) and cytotoxic T lymphocyte antigen-4 (CTLA-4) as first-line treatment options for BRAFV600-mutant melanoma. SECOMBIT (NCT02631447) was a randomized, three-arm, noncomparative phase II trial in which patients were randomized to one of two sequences with immunotherapy or targeted therapy first, with a third arm in which an 8-week induction course of targeted therapy followed by a planned switch to immunotherapy was the first treatment. BRAF/MEK inhibitors were encorafenib plus binimetinib and checkpoint inhibitors ipilimumab plus nivolumab. Primary outcome of overall survival was previously reported, demonstrating improved survival with immunotherapy administered until progression and followed by BRAF/MEK inhibition. Here we report 4-year survival outcomes, confirming long-term benefit with first-line immunotherapy. We also describe preliminary results of predefined biomarkers analyses that identify a trend toward improved 4-year overall survival and total progression-free survival in patients with loss-of-function mutations affecting JAK or low baseline levels of serum interferon gamma (IFNy). These long-term survival outcomes confirm immunotherapy as the preferred first-line treatment approach for most patients with BRAFV600-mutant metastatic melanoma, and the biomarker analyses are hypothesis-generating for future investigations of predictors of durable benefit with dual checkpoint blockade and targeted therapy

AtopyReg®, the Prospective Italian Patient Registry for Moderate-to-Severe Atopic Dermatitis in Adults: Baseline Demographics, Disease Characteristics, Comorbidities, and Treatment History

background and objective atopyReg((R)) is a multicenter, prospective, observational, non-profit cohort study on moderate-tosevere atopic dermatitis in adults promoted in 2018 by the Italian society of dermatology and Venereology (SIDeMaST). we aimed to describe baseline demographics, disease characteristics, comorbidities, and therapeutic data of adult patients affected by moderate-to-severe atopic dermatitis.methods patients were selected based on the following inclusion criteria: age &gt;= 18 years; eczema area and severity Index score &gt;= 7 or a numeric rating scale sleep loss score = 7, or a dermatology life quality Index score &gt;= 10. score &gt;= 16 or localization in visible or sensitive areas (face, neck, hands, or genitalia), or a numeric rating scale itchDemographic and clinical data at baseline were recorded and analyzed. results a total of 1170 patients (male 51.1%; mean age: 44.7 years; range 18-90 years) were enrolled by 12 Italian dermatology units between January 2019 and november 2022. skin lesions were eczematous in 83.2% of patients, the most involved site were the flexures (53.9%), face (50.9%), and neck (48.0%). mean eczema area and severity Index score was 22.3, mean dermatology life quality Index value was 17.6, mean patient oriented Eczema measure score was 13.1, and mean numeric rating scale itch and sleep loss scores were 7.6 and 5.9, respectively. previous systemic therapies were corticosteroids in 77.7% of patients, antihistamines in 50.3% of patients, and cyclosporine A in 42.6% of patients.conclusions this baseline data analysis deriving from atopyReg((R)) provides real-life evidence on patients with moderate-to-severe atopic dermatitis in Italy confirming the high burden of atopic dermatitis with a significant impact on patients' quality of life

Evidence for Sub-Haplogroup H5 of Mitochondrial DNA as a Risk Factor for Late Onset Alzheimer's Disease

BACKGROUND: Alzheimer's Disease (AD) is the most common neurodegenerative disease and the leading cause of dementia among senile subjects. It has been proposed that AD can be caused by defects in mitochondrial oxidative phosphorylation. Given the fundamental contribution of the mitochondrial genome (mtDNA) for the respiratory chain, there have been a number of studies investigating the association between mtDNA inherited variants and multifactorial diseases, however no general consensus has been reached yet on the correlation between mtDNA haplogroups and AD. METHODOLOGY/PRINCIPAL FINDINGS: We applied for the first time a high resolution analysis (sequencing of displacement loop and restriction analysis of specific markers in the coding region of mtDNA) to investigate the possible association between mtDNA-inherited sequence variation and AD in 936 AD patients and 776 cognitively assessed normal controls from central and northern Italy. Among over 40 mtDNA sub-haplogroups analysed, we found that sub-haplogroup H5 is a risk factor for AD (OR=1.85, 95% CI:1.04-3.23) in particular for females (OR=2.19, 95% CI:1.06-4.51) and independently from the APOE genotype. Multivariate logistic regression revealed an interaction between H5 and age. When the whole sample is considered, the H5a subgroup of molecules, harboring the 4336 transition in the tRNAGln gene, already associated to AD in early studies, was about threefold more represented in AD patients than in controls (2.0% vs 0.8%; p=0.031), and it might account for the increased frequency of H5 in AD patients (4.2% vs 2.3%). The complete re-sequencing of the 56 mtDNAs belonging to H5 revealed that AD patients showed a trend towards a higher number (p=0.052) of sporadic mutations in tRNA and rRNA genes when compared with controls. CONCLUSIONS: Our results indicate that high resolution analysis of inherited mtDNA sequence variation can help in identifying both ancient polymorphisms defining sub-haplogroups and the accumulation of sporadic mutations associated with complex traits such as AD