P02.185. The effects of tactile massage (TM) on blood pressure, heart rate and blood glucose in a sample of women suffering from primary insomnia

The overall objective of this pilot study was to study the direct effects of tactile massage (TM) on blood pressure, heart rate and blood glucose in a sample of women suffering from primary insomniaThe study had an experimental prospective design, with a total of 10 women (mean age; 53 years, ±5.4). The participants underwent TM twice a week for six weeks resulting in a total of 120 treatments. For short term effects, systolic and diastolic blood pressure, heartrate and blood glucoses were assessed by the therapist before and after each treatment. Long term assessments were made at baseline, at week 7, and at week 13.As a short term result after the treatment with TM, the participants reached a statistically significant reduction of; systolic blood pressure (-5.5 mmHg, ± 5.0), diastolic blood pressure (-2.0 mmHg, ± 4.4), Heartrate (-5.1 beats per minute, ± 3.4) and blood glucose (-0.2 mmol, ± 0.5). No long term effects with respect to the studied variables can be observed.In summary, we have shown in a normotensive but highly stressed sample of women, that TM has beneficiary effects on parameters of stress and cardiovascular function. In total, 120 TM treatments was analyzed with respect to the objective of the study, but in order to more understand the practical effects, and to more deeply evaluate TM’s place in the modalities of stress reduction, we recommend further studies with larger samples

IL-10 underlies distinct susceptibility of BALB/c and C57BL/6 mice to Mycobacterium avium infection and influences efficacy of antibiotic therapy

Increased production of IL-10 has been frequently associated with augmented susceptibility to infection. However, the correlation between IL-10 activity and susceptibility to mycobacterial infection is still uncertain. Although studies using transgenic mice overexpressing IL-10 consistently showed an increased susceptibility to mycobacterial infection, experimental approaches in which IL-10 activity was reduced or abrogated originated inconclusive data. We show here that this controversy might be due to the mouse strains used in the various experimental procedures. Our results show that BALB/c mice are more susceptible than C57BL/6 to Mycobacterium avium infection. This increased susceptibility of BALB/c mice is, to a great extent, due to distinct activity of IL-10 between the two mouse strains. In accordance, reduction of IL-10 activity through the administration of anti-IL-10R mAb, or the absence of IL-10 as studied in IL-10 knockout mice, clearly decreased the susceptibility of BALB/c mice to M. avium but had a less obvious effect in C57BL/6 mice. Moreover, abrogation of IL-10 activity in infected BALB/c mice increased the efficacy of antimycobacterial therapy, whereas for the C57BL/6 mice it produced no effect. These observations show that the activity of IL-10 in response to the same mycobacterial stimulus influences not only the susceptibility to infection but also the efficacy of antimycobacterial therapy. This should now be considered in the context of human response to mycobacterial infection, particularly as a possible strategy to improve treatment against infections by mycobacteria

Biomedical applications of human cathelicidin

[Excerpt] Antimicrobial peptides (AMPs) are good candidates to treat burn wounds, a major cause of morbidity, impaired life quality and resources consumption in developed countries. Tuberculosis (TB), a disease caused by the human pathogen Mycobacterium tuberculosis, represents the second world’s deadliest infectious disease, affecting around 9 million people worldwide in 2013. Of those, about 1.1 million died from the disease. The potential of cathelicin, a human AMP, in the treatment of mycobacteriosis and wound regeneration was assessed in pre-clinical trials. (...

Contribution of CD30/CD153 but not of CD27/CD70, CD134/OX40L, or CD137/4-1BBL to the optimal induction of protective immunity to Mycobacterium avium

A panel of monoclonal antibodies specific for CD27 ligand (CD70), CD30 ligand (CD153), CD134 ligand (OX40L), and CD137 ligand (4-1BBL) were screened in vivo for their ability to affect the control of Mycobacterium avium infection in C57B1/6 mice. Only the blocking of CD153 led to increased mycobacterial burdens. We then used CD30-deficient mice and found an increase in the proliferation of two strains of M. avium in these mice as compared with control animals. The increased mycobacterial growth was associated with decreased T cell expansion and reduced interferon-gamma (IFN-gamma) responses as a result of reduced polarization of the antigen-specific, IFN-gamma-producing T cells. At late times but not early in infection, the lymphoid cuff surrounding granulomas was depleted in the CD30-deficient animals. This report expands our knowledge about tumor necrosis factor superfamily members involved in the immune responses to mycobacterial infection by identifying CD30-CD153 interactions as required for optimal immune control of M. avium infection

Endogenous cathelicidin production limits inflammation and protective immunity to Mycobacterium avium in mice

The production of antimicrobial peptides, such as the cathelicidins, plays a prominent role in the innate immune response against microbial pathogens. Cathelicidins are widely distributed amongst living organisms, and the antimicrobial peptides generated by proteolysis of the precursor forms are typically cationic and [alpha]-helical, a structure that facilitates their interaction and insertion into anionic bacterial cell walls and membranes, causing damage and promoting microbial death. Here, we found that mouse cathelicidin (Camp) expression was induced in bone marrow-derived macrophages by infection with Mycobacterium avium in a TLR2- and TNF-dependent manner. However, the endogenous production of the cathelin-related antimicrobial peptide (CRAMP) was not required for the bacteriostasis of M. avium either in primary cultures of macrophages or in vivo, as shown by the use of CRAMP-null mice. In contrast, the lack of Camp led to a transient improvement of M. avium growth control in the spleens of infected mice while at the same time causing an exacerbation of the inflammatory response to infection. Our data highlight the anti-inflammatory effects of CRAMP and suggests that virulent mycobacteria may possess strategies to escape its antimicrobial activity.Funded through project PTDC/BIA-BCM/112138/2009FCOMP-01-0124-FEDER014185

Delivery of antimicrobial peptides for the treatment of mycobacteriosis

Mycobacterium tuberculosis, which resides inside macrophages, has always been recognized as one of the most “successful” pathogens. Standard treatments have already been used for decades and, therefore, resistances to the first-line medicines are increasing. Additionally, poor patient compliance with stringent therapies is often pointed out as a major reason leading to treatment failure. Antimicrobial peptides (AMPs), a promising new class of broad spectrum antibiotics, are less prone to result in pathogen resistances due to their target (cellular membranes) and rapid action. In our laboratory we search for AMPs with potent activity against mycobacteria and try to develop efficient delivery systems based on self-assembled colloidal nanocarriers. Additionally, this systems are expected to reduce peptide toxicity and enhance selective uptake on infected cells. Finally, the use of encapsulated drugs in mycobacterial therapy may help reducing drug administration schedules which would ultimately improve patient compliance

Dissemination of mycobacteria to the thymus renders newly generated T cells tolerant to the invading pathogen

The ability of the thymus to generate a population of T cells that is, for the most part, self-restricted and self-tolerant depends to a great extent on the Ags encountered during differentiation. We recently showed that mycobacteria disseminate to the thymus, which raised the questions of how mycobacteria within the thymus influence T cell differentiation and whether such an effect impacts host-pathogen interactions. Athymic nude mice were reconstituted with thymic grafts from Mycobacterium avium-infected or control noninfected donors. T cells generated from thymi of infected donors seemed generally normal, because they retained the ability to reconstitute the periphery and to respond to unspecific stimuli in vitro as well as to antigenic stimulation with third-party Ags, such as OVA, upon in vivo immunization. However, these cells were unable to mount a protective immune response against a challenge with M. avium. The observation that thymic infection interferes with T cell differentiation, generating T cells that are tolerant to pathogen-specific Ags, is of relevance to understand the immune response during chronic persistent infections. In addition, it has potential implications for the repertoire of T cells generated in patients with a mycobacterial infection recovering from severe lymphopenia, such as patients coinfected with HIV and receiving antiretroviral therapy.Fundação para a Ciência e Tecnologia and Fundo Europeu de Desenvolvimento Regional (PIC/IC/83313/2007; PTDC/SAU-MII/101663/2008) and the American-Portuguese Biomedical Research Fund. C.N., S.R., and C.N.-A. are recipients of PhD fellowships from Fundação para a Ciência e Tecnologi

Delivery of nanogel formulations with antimicrobial peptides for the treatment of mycobacteriosis

Book of Abstracts of CEB Annual Meeting 2017[Excerpt] Mycobacterium tuberculosis is the human pathogen that causes Tuberculosis (TB). In 2015, 10.4 million TB cases and 1.8 million deaths were reported, placing this disease alongside HIV/AIDS as the deadliest infectious diseases. Current treatments rely in the administration of a cocktail of four first-line antibiotics during 6 months and, in the worst case scenario, a long-lasting treatment (24 months) with second-line drugs. The overuse or misuse of antimicrobial agents decreases the success of treatments and increases emergence of Multi-drug resistant (MDR) strains. Therefore, the development of new strategies for TB therapy is urgently needed. In this scope, antimicrobial peptides (AMPs) arise as promising candidates for TB treatment since they present high spectrum of antimicrobial activity, high efficacy at low concentrations and low propensity for bacterial resistance. Nevertheless, the low capacity of AMPs to reach the infected site and the use of high concentrations to overcome this problem limits its clinical application - this can be circumvented using a drug delivery system [1]. [...]info:eu-repo/semantics/publishedVersio

Delivery of LLKKK18 loaded into self-assembling hyaluronic acid nanogel for tuberculosis treatment

uberculosis (TB), a disease caused by the human pathogen Mycobacterium tuberculosis, recently joined HIV/AIDS on the top rank of deadliest infectious diseases. Low patient compliance due to the expensive, long-lasting and multi-drug standard therapies often results in treatment failure and emergence of multi-drug resistant strains. In this scope, antimicrobial peptides (AMPs) arise as promising candidates for TB treatment. Here we describe the ability of the exogenous AMP LLKKK18 to efficiently kill mycobacteria. The peptide's potential was boosted by loading into self-assembling Hyaluronic Acid (HA) nanogels. These provide increased stability, reduced cytotoxicity and degradability, while potentiating peptide targeting to main sites of infection. The nanogels were effectively internalized by macrophages and the peptide presence and co-localization with mycobacteria within host cells was confirmed. This resulted in a significant reduction of the mycobacterial load in macrophages infected in vitro with the opportunistic M. avium or the pathogenic M. tuberculosis, an effect accompanied by lowered pro-inflammatory cytokine levels (IL-6 and TNF-). Remarkably, intra-tracheal administration of peptide-loaded nanogels significantly reduced infection levels in mice infected with M. avium or M. tuberculosis, after just 5 or 10 every other day administrations. Considering the reported low probability of resistance acquisition, these findings suggest a great potential of LLKKK18-loaded nanogels for TB therapeutics.This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/ BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER- 006684). The authors also acknowledge the Project RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462). The authors thank Dr. Hugo Osório (Proteomics Lab at I3S – Institute for Health Research and Innovation, Porto, Portugal) for the MALDI-ToF analysis. JPS acknowledges FCT for the financial support provided by grant SFRH/BPD/64958/2010

Characterization of the binding site of the histamine H3 receptor. 1. Various approaches to the synthesis of 2-(1H-imidazol-4-yl)cyclopropylamine and histaminergic activity of (1R,2R)-and (1S,2S)-2-(1H-imidazol-4-yl)-cyclopropylamine.

Various approaches to the synthesis of all four stereoisomers of 2-(1H- imidazol-4-yl)cyclopropylamine (cyclopropylhistamine) are described. The rapid and convenient synthesis and resolution of trans-cyclopropylhistamine is reported. The absolute configuration of its enantiomers was determined by single-crystal X-ray crystallographic analysis. The distinct transcyclopropylhistamine enantiomers were tested for their activity and affinity on the histamine