Confocal laser scanning microscope, raman microscopy and western blotting to evaluate inflammatory response after myocardial infarction

Cardiac muscle necrosis is associated with inflammatory cascade that clears the infarct from dead cells and matrix debris, and then replaces the damaged tissue with scar, through three overlapping phases: the inflammatory phase, the proliferative phase and the maturation phase. Western blotting, laser confocal microscopy, Raman microscopy are valuable tools for studying the inflammatory response following myocardial infarction both humoral and cellular phase, allowing the identification and semiquantitative analysis of proteins produced during the inflammatory cascade activation and the topographical distribution and expression of proteins and cells involved in myocardial inflammation. Confocal laser scanning microscopy (CLSM) is a relatively new technique for microscopic imaging, that allows greater resolution, optical sectioning of the sample and three-dimensional reconstruction of the same sample. Western blotting used to detect the presence of a specific protein with antibody-antigen interaction in the midst of a complex protein mixture extracted from cells, produced semi-quantitative data quite easy to interpret. Confocal Raman microscopy combines the three-dimensional optical resolution of confocal microscopy and the sensitivity to molecular vibrations, which characterizes Raman spectroscopy. The combined use of western blotting and confocal microscope allows detecting the presence of proteins in the sample and trying to observe the exact location within the tissue, or the topographical distribution of the same. Once demonstrated the presence of proteins (cytokines, chemokines, etc.) is important to know the topographical distribution, obtaining in this way additional information regarding the extension of the inflammatory process in function of the time stayed from the time of myocardial infarction. These methods may be useful to study and define the expression of a wide range of inflammatory mediators at several different timepoints providing a more detailed analysis of the time course of the infarct

The meaning of different forms of structural myocardial injury, immune response and timing of infarct necrosis and cardiac repair

Although a decline in the all-cause and cardiac mortality rates following myocardial infarction (MI) during the past 3 decades has been reported, MI is a major cause of death and disability worldwide. From a pathological point of view MI consists in a particular myocardial cell death due to prolonged ischemia. After the onset of myocardial ischemia, cell death is not immediate, but takes a finite period of time to develop. Once complete myocytes’ necrosis has occurred, a process leading to a healed infarction takes place. In fact, MI is a dynamic process that begins with the transition from reversible to irreversible ischemic injury and culminates in the replacement of dead myocardium by a fibrous scar. The pathobiological mechanisms underlying this process are very complex, involving an inflammatory response by several pathways, and pose a major challenge to ability to improve our knowledge. An improved understanding of the pathobiology of cardiac repair after MI and further studies of its underlying mechanisms provide avenues for the development of future strategies directed toward the identification of novel therapies. The chronologic dating of MI is of great importance both to clinical and forensic investigation, that is, the ability to create a theoretical timeline upon which either clinicians or forensic pathologists may increase their ability to estimate the time of MI. Aging of MI has very important practical implications in clinical practice since, based on the chronological dating of MI, attractive alternatives to solve therapeutic strategies in the various phases of MI are developing

Adults and nest of<i>Liostenogaster pardii</i>n. sp. (Hymenoptera Stenogastrinae)

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Unrelated Helpers in a Primitively Eusocial Wasp: Is Helping Tailored Towards Direct Fitness?

The paper wasp Polistes dominulus is unique among the social insects in that nearly one-third of co-foundresses are completely unrelated to the dominant individual whose offspring they help to rear and yet reproductive skew is high. These unrelated subordinates stand to gain direct fitness through nest inheritance, raising the question of whether their behaviour is adaptively tailored towards maximizing inheritance prospects. Unusually, in this species, a wealth of theory and empirical data allows us to predict how unrelated subordinates should behave. Based on these predictions, here we compare helping in subordinates that are unrelated or related to the dominant wasp across an extensive range of field-based behavioural contexts. We find no differences in foraging effort, defense behaviour, aggression or inheritance rank between unrelated helpers and their related counterparts. Our study provides no evidence, across a number of behavioural scenarios, that the behaviour of unrelated subordinates is adaptively modified to promote direct fitness interests

Antifungals, arthropods and antifungal resistance prevention: lessons from ecological interactions

Arthropods can produce a wide range of antifungal compounds, including specialist proteins, cuticular products, venoms and haemolymphs. In spite of this, many arthropod taxa, particularly eusocial insects, make use of additional antifungal compounds derived from their mutualistic association with microbes. Because multiple taxa have evolved such mutualisms, it must be assumed that, under certain ecological circumstances, natural selection has favoured them over those relying upon endogenous antifungal compound production. Further, such associations have been shown to persist versus specific pathogenic fungal antagonists for more than 50 million years, suggesting that compounds employed have retained efficacy in spite of the pathogens' capacity to develop resistance. We provide a brief overview of antifungal compounds in the arthropods' armoury, proposing a conceptual model to suggest why their use remains so successful. Fundamental concepts embedded within such a model may suggest strategies by which to reduce the rise of antifungal resistance within the clinical milieu

Nandrolone decanoate interferes with testosterone biosynthesis altering blood-testis barrier components

The aim of this study was to investigate whether nandrolone decanoate (ND) use affects testosterone production and testicular morphology in a model of trained and sedentary mice. A group of mice underwent endurance training while another set led a sedentary lifestyle and were freely mobile within cages. All experimental groups were treated with either ND or peanut oil at different doses for 6 weeks. Testosterone serum levels were measured via liquid chromatography-mass spectrometry. Western blot analysis and quantitative real-time PCR were utilized to determine gene and protein expression levels of the primary enzymes implicated in testosterone biosynthesis and gene expression levels of the blood-testis barrier (BTB) components. Immunohistochemistry and immunofluorescence were conducted for testicular morphological evaluation. The study demonstrated that moderate to high doses of ND induced a diminished serum testosterone level and altered the expression level of the key steroidogenic enzymes involved in testosterone biosynthesis. At the morphological level, ND induced degradation of the BTB by targeting the tight junction protein-1 (TJP1). ND stimulation deregulated metalloproteinase-9, metalloproteinase-2 (MMP-2) and the tissue inhibitor of MMP-2. Moreover, ND administration resulted in a mislocalization of mucin-1. In conclusion, ND abuse induces a decline in testosterone production that is unable to regulate the internalization and redistribution of TJP1 and may induce the deregulation of other BTB constituents via the inhibition of MMP-2. ND may well be considered as both a potential inducer of male infertility and a potential risk factor to a low endogenous bioavailable testosterone

"PROXIMITY" AS A DESIGN STRATEGY FOR SUSTAINABLE, COLLABORATIVE AND INCLUSIVE URBAN PUBLIC SPACES

Public spaces at neighbourhood scale represent the main scenario of citizens’ life, nodes that define an archipelago of places with a key role in promoting and fostering the enhancement and maintenance of the built environment through mutual collaboration. This network of open and built spaces traces the reference infrastructure of urban planning and redevelopment models, based on proximity as device for physical and social relationship, central in the post-pandemic city debate. “The city of proximity” is the first focus addressed by the Atelier of Urban Innovation Lab in Bologna: an exhibition and laboratory space for comparison and co-design of public space, housed inside the City Hall of the Italian metropolitan city of Bologna and co-curated by the Department of Architecture, University of Bologna and the city agency Foundation for the Urban Innovation. The Atelier explores urban dynamics and contemporary challenges using analogue and digital tools investigating the potential of the ecosystem defining the realm of the everyday dynamics – for instance, squares, courtyards, markets, libraries, urban gardens, playgrounds, sport equipment, etc. In addition to the physical transformations linked to the concept of proximity, the design process, the actors involved and the results obtained are considered relevant. This paper, framing the topic on transition city issues, addresses the potential, transversal andrecurrent features of neighborhood spaces with the aim of acknowledging replicable strategies and design practices for collaborative cities based on new form of citizen democracy that encourage the reactivation of places, community creation, resilience to climate change and sustainable mobility, as experimented by the lab-like environment of Urban Innovation Lab Bologna

The model of “informed refusal” for vaccination: How to fight against anti-vaccinationist misinformation without disregarding the principle of self-determination

Vaccines are arguably a public health success story as well as an incredibly cost-effective medical resource. Despite this, worldwide concerns about their safety are growing, with the risk of increased morbidity and mortality in vaccine-preventable diseases because of vaccine refusal. The global political trend in developed countries is to increasingly reduce mandates and the compulsory nature of vaccination programs. This is due to strong opposition from anti-vaccination movements and groups. While these have existed since the beginnings of vaccinology, they have recently gained a strong foothold through massive exploitation of the media and especially the internet. This has led to widespread misinformation and greater difficulty for governments and health institutions in dealing with parents’ concerns and misconceptions. Common strategies in order to maintain a high degree of public acceptance of vaccines include the enhancement of adverse effect reporting systems, the enrichment of scientific literature, and the dissemination of targeted information to parents and health care providers. Vaccine risk perception, in fact, largely exceeds the evidence and is linked to well-known general population cognitive bias, which must be recognized and corrected. Although there is no doubt about the convenience of universal vaccination, a lively international debate is underway with regard to the legitimacy of mandatory vaccination programs. Most scientists agree that the individual’s right to self-determination should be preserved. The only way to simultaneously protect the right to health is to introduce an informed refusal model, which aims to guarantee the highest coverage rates for vaccination

Kill rate as a tool in efficiency evaluation of Neoseiulus californicus (Acari: Phytoseiidae) mass reared on factitious food

The predatory mites of the Phytoseiidae family are crucial biological control agents widely utilized in biological pest management targeting phytophagous mites and insects. Key factors in these control strategies are that phytoseiids must be able to find their main target prey and to maintain high populations and efficacy. To reduce expenses and time-consuming production methods of mass rearing of phytoseiids, pollen and other factitious (i.e., non-natural/nontarget) hosts need to be present as an alternative food for predatory mite populations. The mass-rearing possibilities of these predators on alternative food sources, such as astigmatid mites (i.e., house and stored mites) and pollen, must be evaluated not only by the cost of rearing settings but on the basis of their efficiency maintenance in killing prey. The pest kill rate (km) is the average daily lifetime killing of the pest by the natural enemy under consideration and can represent a useful indicator for phytoseiids efficacy to rank them as BCAs on the basis of their effective killing/predation on target prey. In this study, we evidenced that 2 astigmatid mites, Glycyphagus domesticus (De Geer) and Lepidoglyphus destructor (Schrank), and Quercus ilex L. pollen can be successfully adopted as substitute food sources for mass rearing of the phytoseiid Neoseiulus californicus (MgGregor); then, we determined that these populations of BCAs maintained a consistent km at new/first impact on the main target pest, Tetranychus urticae Koch