Particulate Air Pollution, Clock Gene Methylation, and Stroke : Effects on Stroke Severity and Disability

Circadian rhythm disturbances have been consistently associated with the development of several diseases, particularly cardiovascular diseases (CVDs). A central clock in the brain maintains the daily rhythm in accordance with the external environment. At the molecular level, the clock is maintained by \u201cclock genes\u201d, the regulation of which is mainly due to DNA methylation, a molecular mechanism of gene expression regulation, able to react to and be reprogrammed by environmental exposure such as exposure to particulate matter (PM). In 55 patients with a diagnosis of acute ischemic stroke, we showed that PM2.5 exposure experienced before the event influenced clock genes methylation (i.e., circadian locomotor output cycles protein kaput CLOCK, period 2 PER2, cryprochrome 1 CRY1, Neuronal PAS Domain Protein 2 NPAS2), possibly modulating the patient prognosis after the event, as cryptochrome 1 CRY1 and period 1 PER1 methylation levels were associated with the Rankin score. Moreover, if PM2.5 annual average was low, CRY1/CRY2 methylation levels were positively associated with the National Institutes of Health Stroke Scale (NIHSS) score, whereas they were negatively associated if PM2.5 exposure was high. Whether epigenetic changes in clock genes need to be considered as a prognostic marker of stroke or rather a causal agent in stroke development remains to be determined. Further studies are needed to determine the role of clock gene methylation in regulating the response to and recovery after a stroke event

Bioactive properties and functional constituents of Hypericum androsaemum L.: A focus on the phenolic profile

Hypericum androsaemum L. ethanol:water extract acted as a lipid peroxidation inhibitor and free radical scavenger. A marked inhibition of the growth of breast, lung, cervical and hepatocellular human carcinoma cell lines was also observed, whereas no toxicity was shown against non-tumor porcine liver cells (> 400 ?g/mL). The extract was also effective in inhibiting nitric oxide production, as an indicator of the anti-inflammatory potential. The anti-Candida effects varied among different strains of the same species, C. glabrata and C. tropicalis being the most sensible species with an effect directly related with the extract concentrations tested. A significant anti-biofilm formation potential was also observed, namely for C. glabrata and C. tropicalis (biofilm reduction > 90%). 5-O-Caffeoylquinic and 3-O-caffeoylquinic acids were the most abundant phenolic compounds identified in the extract, and might be related with the observed bioactive effects. Nevertheless, future studies should be carried out to obtain dose-response curves of the isolated active compounds, in order to perform further pre-clinically testing to quantify the presence of the most active compounds in the extract.The authors thank the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2013), LSRE (Project UID/EQU/50020/2013), N. Martins (SFRH/BD/87658/2012), S. Silva (SFRH/BPD/109053/2015) and L. Barros (SFRH/BPD/107855/2015) grants. The authors would also like to thank Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Conselho Nacional de Desenvolvimento Científico e Tecnológico (Cnpq), and Fundação Araucária for the financial support received. Flávia Tobaldini-Valerio, acknowledge the financial support of CAPES – Proc. 9469/14-1. The authors also thank FCT for the Strategic Project of the UID/BIO/04469/2013 unit, FCT and European Union funds (FEDER/COMPETE) for the project RECI/BBB-EBI/0179/2012 (FCOMP- 01-0124-FEDER-027462)

Propolis: a potential natural product to fight Candida species infections

Aim: To evaluate the effect of propolis against Candida species planktonic cells and its counterpart's biofilms. Materials & methods: The MIC values, time-kill curves and filamentation form inhibition were determined in Candida planktonic cells. The effect of propolis on Candida biofilms was assessed through quantification of CFUs. Results: MIC values, ranging from 220 to 880 µg/ml, demonstrated higher efficiency on C. albicans and C. parapsilosis than on C. tropicalis cells. In addition, propolis was able to prevent Candida species biofilm's formation and eradicate their mature biofilms, coupled with a significant reduction on C. tropicalis and C. albicans filamentation. Conclusion: Propolis is an inhibitor of Candida virulence factors and represents an innovative alternative to fight candidiasis.The authors thank Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Conselho Nacional de Desenvolvimento Científico e Tecnológico (Cnpq) and Fundação Araucária for the financial support received. Flávia Tobaldini-Valerio acknowledges the financial support of CAPES – Proc. 9469/14-1. The authors also thank FCT for the Strategic Project of the UID/BIO/04469/2013 unit, FCT and European Union funds (FEDER/COMPETE) for the project RECI/BBBEBI/0179/2012 (FCOMP-01-0124-FEDER-027462). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed

Hereditary angioedema: Assessing the hypothesis for underlying autonomic dysfunction

Background Attacks of Hereditary Angioedema due to C1-inhibitor deficiency (Cl-INH-HAE) are often triggered by stressful events/hormonal changes. Objective Our study evaluates the relationship between autonomic nervous system (ANS) and contact/complement system activation. Methods Twenty-three HAE patients (6 males, mean age 47.5 +/- 11.4 years) during remission and 24 healthy controls (8 males, mean age 45.3 +/- 10.6 years) were studied. ECG, beat-by-beat blood pressure, respiratory activity were continuously recorded during rest (10') and 75degrees- head-up tilt (10'). C1-INH, C4, cleaved high molecular weight kininogen (cHK) were assessed; in 16 patients and 11 controls plasma catecholamines were also evaluated. Spectral analysis of heart rate variability allowed extraction of low-(LF) and high-(HF)frequency components, markers of sympathetic and vagal modulation respectively. Results HAE patients showed higher mean systolic arterial pressure (SAP) than controls during both rest and tilt. Tilt induced a significant increase in SAP and its variability only in controls. Although sympathetic modulation (LFnu) increased significantly with tilt in both groups, LF/HF ratio, index of sympathovagal balance, increased significantly only in controls. At rest HAE patients showed higher noradrenaline values (301.4 +/- 132.9 pg/ml vs 210.5 +/- 89.6 pg/ml, p = 0.05). Moreover, in patients tilt was associated with a significant increase in cHK, marker of contact system activation (49.5 +/- 7.5% after T vs 47.1 +/- 7.8% at R, p = 0.01). Conclusions Our data are consistent with altered ANS modulation in HAE patients, i.e. increased sympathetic activation at rest and blunted response to orthostatic challenge. Tilt test-induced increased HK cleavage suggests a link between stress and bradykinin production

Sympatho-Vagal Dysfunction in Patients with End-Stage Lung Disease Awaiting Lung Transplantation

Although the literature demonstrates that cardiac autonomic control (CAC) might be impaired in patients with chronic pulmonary diseases, the interplay between CAC and disease severity in end-stage lung disease has not been studied yet. We investigated the effects of end-stage lung disease on CAC through the analysis of heart rate variability (HRV) among patients awaiting lung transplantation. Forty-nine patients on the waiting list for lung transplantation (LTx; 19 men, age 38 \ub1 15 years) and 49 healthy non-smoking controls (HC; 22 men, age 40 \ub1 16 years) were enrolled in a case-control study at Policlinico Hospital in Milan, Italy. LTx patients were divided into two groups, according to disease severity evaluated by the Lung Allocation Score (LAS). To assess CAC, electrocardiogram (ECG) and respiration were recorded at rest for 10 min in supine position and for 10 min during active standing. Spectral analysis identified low and high frequencies (LF, sympathetic, and HF, vagal). Symbolic analysis identified three patterns, i.e., 0V% (sympathetic) and 2UV% and 2LV% (vagal). Compared to HCs, LTx patients showed higher markers of sympathetic modulation and lower markers of vagal modulation. However, more severely affected LTx patients, compared to less severely affected ones, showed an autonomic profile characterized by loss of sympathetic modulation and predominant vagal modulation. This pattern can be due to a loss of sympathetic rhythmic oscillation and a subsequent prevalent respiratory modulation of heart rate in severely affected patients

Chronic Treatment with Ivabradine Does Not Affect Cardiovascular Autonomic Control in Rats

A low resting heart rate (HR) would be of great benefit in cardiovascular diseases. Ivabradine a novel selective inhibitor of hyperpolarization-activated cyclic nucleotide gated (HCN) channels- has emerged as a promising HR lowering drug. Its effects on the autonomic HR control are little known. This study assessed the effects of chronic treatment with ivabradine on the modulatory, reflex and tonic cardiovascular autonomic control and on the renal sympathetic nerve activity (RSNA). Male Wistar rats were divided in 2 groups, receiving intraperitoneal injections of vehicle (VEH) or ivabradine (IVA) during 7 or 8 consecutive days. Rats were submitted to vessels cannulation to perform arterial blood pressure (AP) and HR recordings in freely moving rats. Time series of resting pulse interval and systolic AP were used to measure cardiovascular variability parameters. We also assessed the baroreflex, chemoreflex and the Bezold-Jarish reflex sensitivities. To better evaluate the effects of ivabradine on the autonomic control of the heart, we performed sympathetic and vagal autonomic blockade. As expected, ivabradine treated rats showed a lower resting (VEH: 362 +/- 16 bpm vs. IVA: 260 +/- 14 bpm, p = 0.0005) and intrinsic HR (VEH: 369 +/- 9 bpm vs. IVA: 326 +/- 11 bpm, p = 0.0146). However, the chronic treatment with ivabradine did not change normalized HR spectral parameters LF (nu) (VEH: 24.2 +/- 4.6 vs. IVA: 29.8 +/- 6.4p > 0.05)HF (nu) (VEH: 75.1 +/- 3.7 vs. IVA: 69.2 +/- 5.8p > 0.05), any cardiovascular reflexes, neither the tonic autonomic control of the HR (tonic sympathovagal indexVEH: 0.91 +/- 0.02 vs. IVA: 0.88 +/- 0.03, p = 0.3494). We performed the AP, HR and RSNA recordings in urethane-anesthetized rats. The chronic treatment with ivabradine reduced the resting HR (VEH: 364 +/- 12 bpm vs. IVA: 207 +/- 11 bpm, p < 0.0001), without affecting RSNA (VEH: 117 +/- 16 vs. IVA: 120 +/- 9 spikes/s, p = 0.9100) and mean arterial pressure (VEH: 70 +/- 4 vs. IVA: 77 +/- 6 mmHg, p = 0.3293). Our results suggest that, in health rats, the long-term treatment with ivabradine directly reduces the HR without changing the RSNA modulation and the reflex and tonic autonomic control of the heart.Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)Fundacao de Amparo a Pesquisa de Minas Gerais (FAPEMIG)Universidade Federal de Ouro Preto (UFOP)Universidade Federal do Triangulo Mineiro (UFTM), BrazilUniv Fed Ouro Preto, Inst Exact & Biol Sci, Dept Biol Sci, Lab Cardiovasc Physiol, Ouro Preto, BrazilUniv Fed Ouro Preto, CBIOL NUPEB, Grad Program Biol Sci, Ouro Preto, BrazilUniv Fed Minas Gerais, Inst Biol Sci, Dept Physiol & Biophys, Lab Hypertens, Belo Horizonte, MG, BrazilUniv Fed Sao Paulo, Inst Sci & Technol, Biomed Engn Lab, Sao Jose Dos Campos, BrazilUniv Uberaba, Dept Physiol, Uberaba, BrazilUniv Milan, Osped Maggiore Policlin, IRCCS Ca Granda Fdn, Dept Clin Sci & Community Hlth, Milan, ItalyFed Univ Trianaulo Pvlineiro, Inst Biol & Nat Sci, Dept Physiol, Uberaba, BrazilUniv Fed Sao Paulo, Inst Sci & Technol, Biomed Engn Lab, Sao Jose Dos Campos, BrazilCNPq: 400851/2014-8Web of Scienc

Propolis potential activity against Candida tropicalis adhered cells and its biofilms

Objectives: Invasive fungal infections, such as candidiasis, represent a public health problem of major importance, and Candida tropicalis has been highlighted among the main agents of candidiasis. One of the major contributions to C. tropicalis virulence is its versatility in adapting to a variety of different habitats and the formation of surface attached microbial communities known as biofilms. Moreover, from the clinical perspective, the most important feature of Candida biofilms is its role in increasing tolerance to conventional antifungal therapy. This scenario encourages the search for alternative therapies. Natural matrixes, such as propolis, compromise a multitude of bioactive properties, in particular phenolic extracts have evidenced significant antimicrobial properties against a multiple of opportunist invaders, including Candida species. Thus, the main objective of the present work was to evaluate the potential antifungal effect of propolis against Candida tropicalis biofilms. Methods: This study was conducted with four strains of C. tropicalis and one reference strain, from the American Type Culture Collection (ATCC 40042). Biofilm formation were carried out on 96-well microplates containing a cellular suspension of 1x105 cells/mL and incubated for 24 h at 37°C. Pre-formed C. tropicalis biofilms were treated with propolis (ranging from 0.47 to 1.42 mg/ml), during 24 h at 37°C and its effect assessed through quantification of the number of colony forming unit (CFU). Results: It was evident that all C. tropicalis strains tested were able to form biofilm and that propolis was able to reduce around 40% and 50% of the pre-formed biofilm. Moreover, in general the propolis effect was similar among all the C. tropicalis clinical isolates strains Conclusions: These data are promising, since they open important perspectives regarding new antifungal agents, much more effective and safer than the currently available to treat and to prevent C. tropicalis infections

Physiological and autonomic stress responses after prolonged sleep restriction and subsequent recovery sleep in healthy young men

Purpose Sleep restriction is increasingly common and associated with the development of health problems. We investigated how the neuroendocrine stress systems respond to prolonged sleep restriction and subsequent recovery sleep in healthy young men. Methods After two baseline (BL) nights of 8 h time in bed (TIB), TIB was restricted to 4 h per night for five nights (sleep restriction, SR, n = 15), followed by three recovery nights (REC) of 8 h TIB, representing a busy workweek and a recovery weekend. The control group (n = 8) had 8 h TIB throughout the experiment. A variety of autonomic cardiovascular parameters, together with salivary neuropeptide Y (NPY) and cortisol levels, were assessed. Results In the control group, none of the parameters changed. In the experimental group, heart rate increased from 60 +/- 1.8 beats per minute (bpm) at BL, to 63 +/- 1.1 bpm after SR and further to 65 +/- 1.8 bpm after REC. In addition, whole day low-frequency to-high frequency (LF/HF) power ratio of heart rate variability increased from 4.6 +/- 0.4 at BL to 6.0 +/- 0.6 after SR. Other parameters, including salivary NPY and cortisol levels, remained unaffected. Conclusions Increased heart rate and LF/HF power ratio are early signs of an increased sympathetic activity after prolonged sleep restriction. To reliably interpret the clinical significance of these early signs of physiological stress, a follow-up study would be needed to evaluate if the stress responses escalate and lead to more unfavourable reactions, such as elevated blood pressure and a subsequent elevated risk for cardiovascular health problems.Peer reviewe

International Consensus Based Review and Recommendations for Minimum Reporting Standards in Research on Transcutaneous Vagus Nerve Stimulation (Version 2020).

Given its non-invasive nature, there is increasing interest in the use of transcutaneous vagus nerve stimulation (tVNS) across basic, translational and clinical research. Contemporaneously, tVNS can be achieved by stimulating either the auricular branch or the cervical bundle of the vagus nerve, referred to as transcutaneous auricular vagus nerve stimulation(VNS) and transcutaneous cervical VNS, respectively. In order to advance the field in a systematic manner, studies using these technologies need to adequately report sufficient methodological detail to enable comparison of results between studies, replication of studies, as well as enhancing study participant safety. We systematically reviewed the existing tVNS literature to evaluate current reporting practices. Based on this review, and consensus among participating authors, we propose a set of minimal reporting items to guide future tVNS studies. The suggested items address specific technical aspects of the device and stimulation parameters. We also cover general recommendations including inclusion and exclusion criteria for participants, outcome parameters and the detailed reporting of side effects. Furthermore, we review strategies used to identify the optimal stimulation parameters for a given research setting and summarize ongoing developments in animal research with potential implications for the application of tVNS in humans. Finally, we discuss the potential of tVNS in future research as well as the associated challenges across several disciplines in research and clinical practice