Carbon Nanoparticles Inhibit the Antimicrobial Activities of the Human Cathelicidin LL-37 through Structural Alteration.

Host defense peptides, also known as antimicrobial peptides, are key elements of innate host defense. One host defense peptide with well-characterized antimicrobial activity is the human cathelicidin, LL-37. LL-37 has been shown to be upregulated at sites of infection and inflammation and is regarded as one of the primary innate defense molecules against bacterial and viral infection. Human exposure to combustion-derived or engineered nanoparticles is of increasing concern, and the implications of nanomaterial exposure on the human immune response is poorly understood. However, it is widely acknowledged that nanoparticles can interact strongly with several immune proteins of biological significance, with these interactions resulting in structural and functional changes of the proteins involved. This study investigated whether the potent antibacterial and antiviral functions of LL-37 were inhibited by exposure to, and interaction with, carbon nanoparticles, together with characterizing the nature of the interaction. LL-37 was exposed to carbon black nanoparticles in vitro, and the antibacterial and antiviral functions of the peptide were subsequently assessed. We demonstrate a substantial loss of antimicrobial function when the peptide was exposed to low concentrations of nanomaterials, and we further show that the nanomaterial-peptide interaction resulted in a significant change in the structure of the peptide. The human health implications of these findings are significant, as, to our knowledge, this is the first evidence that nanoparticles can alter host defense peptide structure and function, indicating a new role for nanoparticle exposure in increased disease susceptibility.

A proposal to explain how the circatidal rhythm of the Arabidopsis thaliana root elongation rate could be mediated by the lunisolar gravitational force: a quantum physical approach.

Background and Aims: Roots of Arabidopsis thaliana exhibit a 24.8 h oscillation of elongation rate when grown under free-running conditions. This growth rhythm is synchronized with the time course of the local lunisolar tidal acceleration. The present study aims at a physiological/physical model to describe the interaction of weak gravitational fields with cellular water dynamics that mediate rhythmic root growth profiles. Methods: Fundamental physical laws are applied to model the water dynamics within single plant cells in an attempt to mimic the 24.8 h oscillations in root elongation growth. In particular, a quantum gravitational description of the time course in root elongation is presented, central to which is the formation of coherent assemblies of mass due to the lunisolar gravitational field. Mathematical equations that characterize lunisolar gravity-induced coherent assemblies of water molecules are derived and related to the mass of cellular water within roots of A. thaliana. Key Results: The derived physical model of gravitationally modulated water assemblies is capable of accounting for the experimentally observed arabidopsis root growth kinetics under free-running conditions. The close analogy between the derived time-dependent lunisolar effect upon coherent molecular states of water within single cells and the coherent assemblies of electrons that characterize the quantum Hall effect is emphasized. Conclusions: The dynamics of the lunisolar-induced variation in coherent water assemblies provide a possible mechanism to describe the observed 24.8 h oscillation of root growth rate of A. thaliana. Therefore, this mechanism could function as an independent timekeeper to control cell elongation.

Distribution of some pectic and arabinogalactan protein epitopes during Solanum lycopersicum (L.) adventitious root development.

BACKGROUND: The adventitious roots (AR) of plants share the same function as primary and lateral roots (LR), although their development is mainly an adaptive reaction to stress conditions. Regeneration of grafted plants is often accompanied by AR formation thus making the grafting technique a good model for studying AR initiation and development and their means of emergence. Pectins and arabinogalactan proteins (AGP) are helpful markers of particular cellular events, such as programmed cell death (PCD), elongation, proliferation or other differentiation events that accompany AR development. However, little is known about the distribution of pectins and AGPs during AR ontogeny, either in the primordium or stem tissues from which AR arise or their correspondence with these events during LR formation. RESULTS: AR were developed from different stem tissues such as parenchyma, xylem rays and the cambium, depending on the stem age and treatment (grafting versus cutting) of the parental tissue. Immunochemical analysis of the presence of pectic (LM8, LM19, LM20) and AGP (JIM8, JIM13, JIM16) epitopes in AR and AR-associated tissues showed differential, tissue-specific distributions of these epitopes. Two pectic epitopes (LM19, LM20) were developmentally regulated and the occurrence of the LM8 xylogalacturonan epitope in the root cap of the AR differed from other species described so far. AGP epitopes were abundantly present in the cytoplasmic compartments (mainly the tonoplast) and were correlated with the degree of cell vacuolisation. JIM8 and JIM13 epitopes were detected in the more advanced stages of primordium development, whereas the JIM16 epitope was present from the earliest division events of the initial AR cells. The comparison between AR and LR showed quantitative (AGP,) and qualitative (pectins) differences. CONCLUSION: The chemical compositions of adventitious and lateral root cells show differences that correlate with the different origins of these cells. In AR, developmental changes in the distribution of pectins and AGP suggest the turnover of wall compounds. Our data extend the knowledge about the distribution of pectin and AGP during non-embryogenic root development in a species that is important from an agronomic point of view.

Lunar gravity affects leaf movement of Arabidopsis thaliana in the International Space Station.

MAIN CONCLUSION: Cyclic leaf ascent and descent occur in synchrony and phase congruence with the lunisolar tidal force under a broad range of conditions. Digitized records of the vertical leaf movements of Arabidopsis thaliana were collected under space flight conditions in the International Space Station (ISS). Oscillations of leaf movements with periods of 45 and 90 min were found under light-adapted conditions, whereas in darkness, the periods were 45, 90, and 135 min. To demonstrate the close relationship between these oscillations and cyclical variations of the lunisolar gravitational force, we estimated the oscillations of the in-orbit lunisolar tide as they apply to the ISS, with the aid of the Etide software application. In general, in-orbit lunisolar gravitational profiles exhibited a periodicity of 45 min. Alignment of these in-orbit oscillations with the oscillations of Arabidopsis leaf movement revealed high degrees of synchrony and a congruence of phase. These data corroborate previous results which suggested a correlative relationship and a possible causal link between leaf movement rhythms obtained on ground and the rhythmic variation of the lunisolar tidal force.

Leaf movements and their relationship with the lunisolar gravitational force.

BACKGROUND: Observation of the diurnal ascent and descent of leaves of beans and other species, as well as experimental interventions into these movements, such as exposures to light at different times during the movement cycle, led to the concept of an endogenous 'clock' as a regulator of these oscillations. The physiological basis of leaf movement can be traced to processes that modulate cell volume in target tissues of the pulvinus and petiole. However, these elements of the leaf-movement process do not completely account for the rhythms that are generated following germination in constant light or dark conditions, or when plants are transferred to similar free-running conditions. SCOPE: To develop a new perspective on the regulation of leaf-movement rhythms, many of the published time courses of leaf movements that provided evidence for the concept of the endogenous clock were analysed in conjunction with the contemporaneous time courses of the lunisolar tidal acceleration at the relevant experimental locations. This was made possible by application of the Etide program, which estimates, with high temporal resolution, local gravitational changes as a consequence of the diurnal variations of the lunisolar gravitational force due to the orbits and relative positions of Earth, Moon and Sun. In all cases, it was evident that a synchronism exists between the times of the turning points of both the lunisolar tide and of the leaftide when the direction of leaf movement changes. This finding of synchrony leads to the hypothesis that the lunisolar tide is a regulator of the leaftide, and that the rhythm of leaf movement is not necessarily of endogenous origin but is an expression of an exogenous lunisolar 'clock' impressed upon the leaf-movement apparatus. CONCLUSIONS: Correlation between leaftide and Etide time courses holds for leaf movement rhythms in natural conditions of the greenhouse, in conditions of constant light or dark, under microgravity conditions of the International Space Station, and also holds for rhythms that are atypical, such as pendulum and relaxation rhythms whose periods are longer or shorter than usual. Even the apparently spontaneous short-period, small-amplitude rhythms recorded from leaves under unusual growth conditions are consistent with the hypothesis of a lunisolar zeitgeber. Two hypotheses that could account for the synchronism between leaftide and Etide, and which are based on either quantum considerations or on classical Newtonian physics, are presented and discussed.

Fluorescence decay of dyed protozoa: differences between stressed and non-stressed cysts.

Several series of tests have shown that fresh, intact samples of Giardia duodenalis and Cryptosporidium parvum (oo)cysts are not marked by fluorescent probes such as carboxyfluorcein-succinimidyl-diacetate-ester (CFDA-SE), C12-resazurin and SYTOX® Green, probably because of their robust cell walls. These dyes fail to indicate the viability of such protozoa and allow negative responses to be recorded from living and infectious samples. Cryptosporidium parvum showed stronger isolation from chemicals, with living oocysts remaining unstained by the probe for up to 90 days after extraction. However, in further fluorescence decay (FD) experiments run with G. duodenalis samples stained using CFDA-SE (comprising living, non-stressed but aged cysts, heat-killed samples and UV-C-stressed samples) each showed a different FD decay profile, here studied in seven series of tests of five replicates each. The FD profiles were fitted by double-exponential decay kinetics, with the decay constant k2 being five times higher than k1. This FD procedure is fast and can be easily reproduced in 10 steps, taking ~ 1 h of laboratory work for already purified samples.

Arabidopsis thaliana root elongation growth is sensitive to lunisolar tidal acceleration and may also be weakly correlated with geomagnetic variations.

BACKGROUND: Correlative evidence suggests a relationship between the lunisolar tidal acceleration and the elongation rate of arabidopsis roots grown under free-running conditions of constant low light. METHODS: Seedlings of Arabidopsis thaliana were grown in a controlled-climate chamber maintained at a constant temperature and subjected to continuous low-level illumination from fluorescent tubes, conditions that approximate to a 'free-running' state in which most of the abiotic factors that entrain root growth rates are excluded. Elongation of evenly spaced, vertical primary roots was recorded continuously over periods of up to 14 d using high temporal- and spatial-resolution video imaging and were analysed in conjunction with geophysical variables. KEY RESULTS AND CONCLUSIONS: The results confirm the lunisolar tidal/root elongation relationship. Also presented are relationships between the hourly elongation rates and the contemporaneous variations in geomagnetic activity, as evaluated from the disturbance storm time and ap indices. On the basis of time series of root elongation rates that extend over ≥4 d and recorded at different seasons of the year, a provisional conclusion is that root elongation responds to variation in the lunisolar force and also appears to adjust in accordance with variations in the geomagnetic field. Thus, both lunisolar tidal acceleration and the geomagnetic field should be considered as modulators of root growth rate, alongside other, stronger and more well-known abiotic environmental regulators, and perhaps unexplored factors such as air ions. Major changes in atmospheric pressure are not considered to be a factor contributing to oscillations of root elongation rate.

Area-level deprivation, neighbourhood factors and associations with mental health.

The COVID-19 pandemic saw residential neighbourhoods become more of a focal point in people's lives, where people were greater confined to living, working, and undertaking leisure in their locality. This study investigates whether area-level deprivation and neighbourhood conditions influence mental health, accounting for demographic, socio-economic and health circumstances of individuals. Using nationally representative data from Ireland, regression modelling revealed that area-level deprivation did not in itself have a discernible impact on mental health status (as measured using the Mental Health Inventory-5 instrument and the Energy and Vitality Index), or likelihood of having suffered depression in the previous 12 months. However, positive perceptions of area safety, service provision, and area cleanliness were associated with better mental health, as was involvement in social groups. Broad ranging policies investing in neighbourhoods, could have benefits for mental health, which may be especially important for deprived communities.

Evolution and immunopathology of chikungunya virus informs therapeutic development.

Chikungunya virus (CHIKV), a mosquito-borne alphavirus, is an emerging global threat identified in more than 60 countries across continents. The risk of CHIKV transmission is rising due to increased global interactions, year-round presence of mosquito vectors, and the ability of CHIKV to produce high host viral loads and undergo mutation. Although CHIKV disease is rarely fatal, it can progress to a chronic stage, during which patients experience severe debilitating arthritis that can last from several weeks to months or years. At present, there are no licensed vaccines or antiviral drugs for CHIKV disease, and treatment is primarily symptomatic. This Review provides an overview of CHIKV pathogenesis and explores the available therapeutic options and the most recent advances in novel therapeutic strategies against CHIKV infections.

Arabidopsis thaliana root growth kinetics and lunisolar tidal acceleration.

• All living organisms on Earth are continually exposed to diurnal variations in the gravitational tidal force due to the Sun and Moon. • Elongation of primary roots of Arabidopsis thaliana seedlings maintained at a constant temperature was monitored for periods of up to 14 d using high temporal- and spatial-resolution video imaging. The time-course of the half-hourly elongation rates exhibited an oscillation which was maintained when the roots were placed in the free-running condition of continuous illumination. • Correlation between the root growth kinetics collected from seedlings initially raised under several light protocols but whose roots were subsequently in the free-running condition and the lunisolar tidal profiles enabled us to identify that the latter is the probable exogenous determinant of the rhythmic variation in root elongation rate. Similar observations and correlations using roots of Arabidopsis starch mutants suggest a central function of starch metabolism in the response to the lunisolar tide. The periodicity of the lunisolar tidal signal and the concomitant adjustments in root growth rate indicate that an exogenous timer exists for the modulation of root growth and development. • We propose that, in addition to the sensitivity to Earthly 1G gravity, which is inherent to all animals and plants, there is another type of responsiveness which is attuned to the natural diurnal variations of the lunisolar tidal force.

Lunisolar tidal force and the growth of plant roots, and some other of its effects on plant movements.

BACKGROUND: Correlative evidence has often suggested that the lunisolar tidal force, to which the Sun contributes 30 % and the Moon 60 % of the combined gravitational acceleration, regulates a number of features of plant growth upon Earth. The time scales of the effects studied have ranged from the lunar day, with a period of approx. 24.8 h, to longer, monthly or seasonal variations. SCOPE: We review evidence for a lunar involvement with plant growth. In particular, we describe experimental observations which indicate a putative lunar-based relationship with the rate of elongation of roots of Arabidopsis thaliana maintained in constant light. The evidence suggests that there may be continuous modulation of root elongation growth by the lunisolar tidal force. In order to provide further supportive evidence for a more general hypothesis of a lunisolar regulation of growth, we highlight similarly suggestive evidence from the time courses of (a) bean leaf movements obtained from kymographic observations; (b) dilatation cycles of tree stems obtained from dendrograms; and (c) the diurnal changes of wood-water relationships in a living tree obtained by reflectometry. CONCLUSIONS: At present, the evidence for a lunar or a lunisolar influence on root growth or, indeed, on any other plant system, is correlative, and therefore circumstantial. Although it is not possible to alter the lunisolar gravitational force experienced by living organisms on Earth, it is possible to predict how this putative lunisolar influence will vary at times in the near future. This may offer ways of testing predictions about possible Moon-plant relationships. As for a hypothesis about how the three-body system of Earth-Sun-Moon could interact with biological systems to produce a specific growth response, this remains a challenge for the future. Plant growth responses are mainly brought about by differential movement of water across protoplasmic membranes in conjunction with water movement in the super-symplasm. It may be in this realm of water movements, or even in the physical forms which water adopts within cells, that the lunisolar tidal force has an impact upon living growth systems.

Spontaneous ultra-weak light emissions from wheat seedlings are rhythmic and synchronized with the time profile of the local gravimetric tide.

Semi-circadian rhythms of spontaneous photon emission from wheat seedlings germinated and grown in a constant environment (darkened chamber) were found to be synchronized with the rhythm of the local gravimetric (lunisolar) tidal acceleration. Time courses of the photon-count curves were also found to match the growth velocity profile of the seedlings. Pair-wise analyses of the data--growth, photon count, and tidal--by local tracking correlation always revealed significant coefficients (P > 0.7) for more than 80% of any of the time periods considered. Using fast Fourier transform, the photon-count data revealed periodic components similar to those of the gravimetric tide. Time courses of biophoton emissions would appear to be an additional, useful, and innovative tool in both chronobiological and biophysical studies.

Utilisation of healthcare by immigrant adults relative to the host population: Evidence from Ireland.

OBJECTIVE: While there is a broad consensus that barriers to access in the utilisation of healthcare exist for immigrants in the US, European evidence exploring this issue paints a mixed picture, with studies from a variety of European jurisdictions presenting different conclusions. In this context, Ireland, a European country with substantial private involvement in healthcare delivery, and, a largely young immigrant population, provides an opportunity to investigate the healthcare utilisation of immigrants compared to natives in a European country with mixed private-public healthcare provision. DESIGN: The healthcare utilisation patterns of immigrants (defined as residents with a foreign country of birth) and native-born participants were analysed from a nationally representative health survey of 6,326 adults, carried out in Ireland in 2016. An array of socio-economic and health information was collected such that regression analysis on healthcare consultations accounted for confounding factors. RESULTS: Non-native residents of Ireland born outside the UK were less likely to have attended a General Practitioner (Odds ratio (OR): 0.62 [95% Confidence Interval (CI): 0.51-0.74]; p<0.001) or consultant doctor (OR: 0.60 [95% CI: 0.47-0.76]; p<0.001) in the previous year, relative to Irish-born individuals. UK-born residents of Ireland displayed similar utilisation patterns to those of the native population in terms of GP visitation, but a higher likelihood of having attended a consultant (OR: 1.44 [95% CI: 1.14-1.816]; p = 0.004). CONCLUSIONS: Lower use of healthcare by those born outside Ireland and the UK relative to the native Irish population may be due to different approaches to healthcare utilisation or obstacles to healthcare utilisation. The findings suggest that the utilisation of healthcare by immigrants merits continued policy attention to respond to the needs of these key groups in society and facilitate integration.

Histology and symplasmic tracer distribution during development of barley androgenic embryos.

The present study concerns three aspects of barley androgenesis: (1) the morphology and histology of the embryos during their development, (2) the time course of fluorescent symplasmic tracers' distribution, and (3) the correlation between symplasmic communication and cell differentiation. The results indicate that barley embryos, which are developing via an androgenic pathway, resemble their zygotic counterparts with respect to their developmental stages, morphology and histology. Analysis of the distribution of the symplasmic tracers, HPTS, and uncaged fluorescein indicates the symplasmic isolation of (1) the protodermis from the underlying cells of the late globular stage onwards, and (2) the embryonic organs at the mature stage of development.

Area-level deprivation and geographic factors influencing utilisation of General Practitioner services.

Inequities in access to General Practitioner (GP) services are a key policy concern given the role of GPs as gatekeepers to secondary care services. Geographic or area-level factors, including local deprivation and supply of healthcare providers, are important elements of access. In considering how area-level deprivation relates to GP utilisation, two potentially opposing factors may be important. The supply of healthcare services tends to be lower in areas of higher deprivation. However, poorer health status among individuals in deprived areas suggests greater need for healthcare. To explore the relationship of area-level deprivation to healthcare utilisation, we use data from the Healthy Ireland survey, which provided a sample of 6326 respondents to face-to-face interviews. A u-shaped relationship between GP supply and area-level deprivation is observed in the data. Modelling reveals that residing in more deprived communities has a strong, statistically significant positive association with having seen a GP within the last four weeks, controlling for individual characteristics and GP supply. All else equal, residing in an area ranked in the most deprived quintile increases the odds of a respondent having visited the GP in four weeks by 1.43 (95% Confidence Interval: 1.15-1.78), compared to the least deprived quintile (p-value< 0.001). The findings indicate that the level of deprivation in an area may be relevant to decisions about how to allocate primary care resources.

The human cathelicidin LL-37 preferentially promotes apoptosis of infected airway epithelium.

Cationic host defense peptides are key, evolutionarily conserved components of the innate immune system. The human cathelicidin LL-37 is an important cationic host defense peptide up-regulated in infection and inflammation, specifically in the human lung, and was shown to enhance the pulmonary clearance of the opportunistic pathogen Pseudomonas aeruginosa in vivo by as yet undefined mechanisms. In addition to its direct microbicidal potential, LL-37 can modulate inflammation and immune mechanisms in host defense against infection, including the capacity to modulate cell death pathways. We demonstrate that at physiologically relevant concentrations of LL-37, this peptide preferentially promoted the apoptosis of infected airway epithelium, via enhanced LL-37-induced mitochondrial membrane depolarization and release of cytochrome c, with activation of caspase-9 and caspase-3 and induction of apoptosis, which only occurred in the presence of both peptide and bacteria, but not with either stimulus alone. This synergistic induction of apoptosis in infected cells was caspase-dependent, contrasting with the caspase-independent cell death induced by supraphysiologic levels of peptide alone. We demonstrate that the synergistic induction of apoptosis by LL-37 and Pseudomonas aeruginosa required specific bacteria-epithelial cell interactions with whole, live bacteria, and bacterial invasion of the epithelial cell. We propose that the LL-37-mediated apoptosis of infected, compromised airway epithelial cells may represent a novel inflammomodulatory role for this peptide in innate host defense, promoting the clearance of respiratory pathogens.

Citrullination Alters the Antiviral and Immunomodulatory Activities of the Human Cathelicidin LL-37 During Rhinovirus Infection.

Human rhinoviruses (HRV) are the most common cause of viral respiratory tract infections. While normally mild and self-limiting in healthy adults, HRV infections are associated with bronchiolitis in infants, pneumonia in immunocompromised patients, and exacerbations of asthma and COPD. The human cathelicidin LL-37 is a host defense peptide (HDP) with broad immunomodulatory and antimicrobial activities that has direct antiviral effects against HRV. However, LL-37 is known to be susceptible to the enzymatic activity of peptidyl arginine deiminases (PAD), and exposure of the peptide to these enzymes results in the conversion of positively charged arginines to neutral citrullines (citrullination). Here, we demonstrate that citrullination of LL-37 reduced its direct antiviral activity against HRV. Furthermore, while the anti-rhinovirus activity of LL-37 results in dampened epithelial cell inflammatory responses, citrullination of the peptide, and a loss in antiviral activity, ameliorates this effect. This study also demonstrates that HRV infection upregulates PAD2 protein expression, and increases levels of protein citrullination, including histone H3, in human bronchial epithelial cells. Increased PADI gene expression and HDP citrullination during infection may represent a novel viral evasion mechanism, likely applicable to a wide range of pathogens, and should therefore be considered in the design of therapeutic peptide derivatives.

Interactions Between Autophagy and the Unfolded Protein Response: Implications for Inflammatory Bowel Disease.

Inflammatory bowel disease (IBD), which includes Crohn's disease (CD) and ulcerative colitis, is characterized by chronic inflammation of the gastrointestinal tract. The etiology involves a combination of genetic and environmental factors resulting in abnormal immune responses to intestinal microbiota. Genetic studies have strongly linked genes involved in autophagy to CD, and genes involved in the unfolded protein response (UPR) to IBD. The UPR is triggered in response to accumulation of misfolded proteins in the endoplasmic reticulum (ER), and autophagy plays a key role in relieving ER stress and restoring homeostasis. This review summarizes the known interactions between autophagy and the UPR and discusses the impact of these converging pathways on IBD pathogenesis. With a paucity of effective long-term treatments for IBD, targeting of synergistic pathways may provide novel and more effective therapeutic options.