Epidermolysis bullosa comprises a group of phenotypically differentgenodermatosis, hereditary or acquired, characterized by skin fragility andsubsequent formation of blisters in response to mechanical trauma, andwhich may also affect mucous membranes. This study aimed to analyze therelation between the nutritional, hematologic, infectious characteristicsand the type of epidermolysis bullosa, through a descriptive case studybased on data from medical records of 10 patients with epidermolysisbullosa assisted regularly at the Dermatology Clinic of the HospitalUniversitário de Brasília. The old classification of the type ofepidermolysis bullosa, weight and height, blood count, white blood cellcount, platelet count and description of the type and frequency ofsecondary infections during the service were considered. We verified apredominance of iron deficiency anemia, chronic leukocytosis,thrombocytosis, chronic malnutrition, low height for age and thinness, andpeople with epidermolysis bullosa simplex exhibited appropriate relationbetween height/age and BMI/age. The non-specific skin infection was themost prevalent in both sexes. The severity of the type of epidermolysisbullosa and frequency of secondary infections did not form a directlyproportional relation. The absence of direct proportion in all casesbetween the type of epidermolysis bullosa and the analysis parameterssuggest a possible significant interference from other aspects such as theextent of the affected skin area, extracutaneous type of engagement andspecific genetic mutation. The inclusion of these factors in the newclassification proposed by Fine et al can contribute significantly to abetter correlation of clinical parameters and appropriate preventive andtherapeutic approaches. The old classification of the type of EB, weight and height, blood count,white blood cell count, platelet count and description of the type and frequency ofsecondary infections throughout the service were considered. There was a predominance of chronic malnutrition, low height for age and thinness, andpatients with EB simplex presented appropriate relation between height/age andBMI/age. Also, there was a predominance of cases of iron deficiency anemia, chronicleukocytosis and thrombocytosis without direct correlation to the type of EB.Nonspecific skin infection was the most prevalent in both sexes. Severity of the typeof EB and frequency of secondary infections did not form a directly proportionalrelation.Absence of direct proportion in all cases between the type of EB and analysisparameters suggest a possible significant interference from other aspects such as theextent of the affected skin area, extracutaneous involvement and type of specificgenetic mutation. Inclusion of these factors in the new classification proposed byFine et al can contribute significantly to a better correlation of clinicalparameters and appropriate preventive and therapeutic approaches.Due to the rarity of the disease, the number of cases studied was small and furtherstudies are needed to better evaluate the data found.


Objectives. This systematic review aimed to summarize and evaluate the findings of studies investigating the local microcirculatory effects following acupuncture stimulation. Methods. MEDLINE, EMBASE, OASIS, and Cochrane library were searched to identify randomized controlled trials (RCTs) published before January 30, 2015. Studies demonstrating any type of microcirculation response to manual acupuncture in healthy subjects and patients were included. The risk of bias and the reliability of the experimental conditions were evaluated to determine quality assessment. Results. Eight RCTs met the inclusion criteria; there was at least one acupuncture-induced change in a microcirculatory parameter. Of the seven studies in healthy subjects, four reported significant increases in blood flow following acupuncture compared with control, whereas one other study observed reductions in microcirculation immediately after acupuncture needling. The studies that assessed patients with either fibromyalgia or trapezius myalgia found significant increases in blood flow in the skin and muscle. Additionally, the degree and duration of increases in microcirculation varied depending on the condition of the subjects and the manipulation technique. Conclusions. The current evidence regarding the local effects of acupuncture in terms of blood flow remains insufficient for reliable conclusions due to few well-designed studies. Additional well-designed studies are needed to clarify these issues. Acupuncture is a practical and cost-effective intervention with few adverse side effects that is useful for the treatment of many disorders. In particular, acupuncture treatment has been shown to have analgesic effects in patients with various pain conditions, such as low back pain, headaches, and osteoarthritis . AlthougMicrocirculation is affected by the autonomic nervous system (ANS) and is regarded as a useful method with which to evaluate the peripheral effects of acupuncture –8. In anThus, the primary aims of the present review were to summarize and evaluate the effects of acupuncture on local microcirculation and to compare the blood flow changes induced by acupuncture among five types of typical control groups: no-acupuncture, nonpenetrating acupuncture at same acupoint, superficial acupuncture, acupuncture needling at a nonacupoint, and insertion-only without manipulation.∗” and “blood flow, microcirculation, Doppler, near infrared spectroscopy, near infrared, or perfusion.” Additionally, the reference lists of the selected studies and relevant reviews were also manually searched to identify other appropriate studies.The present review conducted a literature search of the Cochrane library, MEDLINE, EMBASE, and Oriental Medicine Advanced Searching Integrated System (OASIS) databases for studies investigating acupuncture-induced changes in local blood flow that were published up to January 30, 2015. The search was restricted to human studies using the following terms in combination: “acupuncture, ear acupuncture, moxibustion, meridian or acupointThe present review only included original randomized controlled trials (RCTs) that were written in English, Chinese, or German. The details of the inclusion criteria are as follows: (1) the study participants were healthy subjects or patients, (2) the study employed manual needle acupuncture at acupoint(s) and compared the results with a sham control group , insertion-only without manipulation or a subject who did not receive acupuncture, and (3) the outcome included any microcirculatory response to acupuncture including volume, velocity, blood flow, blood flow resistance, and blood vessel diameter around the stimulus sites. Studies were excluded from the present analysis if (1) the intervention included acupuncture-related techniques without skin penetration or did not employ manual acupuncture  and/or (2) the outcome measures did not focus on changes in local blood flow.The data extraction was performed by four reviewers  using a predefined form. Research characteristics such as study design, population, intervention and control groups, outcome measures, and comparisons of the blood flow results between study groups were extracted from each article. Two other reviewers (Hi-Joon Park and Hyangsook Lee) independently verified the extracted data.To assess the methodological quality of the selected studies, two types of checklists were used. First, a quality assessment was performed by two reviewers (Song-Yi Kim and Ji-Yeun Park) using a modified version of the tool for the assessment of risk of bias (RoB) from the Cochrane Handbook . SelectiThe electronic database searches and manual selection of additional experiments identified 697 studies that fit the inclusion criteria. After the exclusion of non-English/Chinese/German-language publications and studies that did not use human subjects based on their titles and abstracts, 315 studies remained eligible for further assessment; of these, eight were RCTs that were ultimately included in the present review .The eight selected RCTs included a total of 205 participants that met the inclusion criteria of the present review; the characteristics of the studies are summarized in In most of the selected studies, acupuncture was performed at points located distal to the elbows and knees such as LI4 , 15, ST3The primary outcome measure was changes in skin or muscle blood flow compared with the control group. To measure the local effects of acupuncture, the measurement sites were stimulated at an acupoint, a region adjacent to the acupoint, or the nearby fingertip. The selected studies assessed outcomes relevant to blood flow, including the blood perfusion ratio, microcirculatory volume, velocity of blood flow, and capillary caliber. The studies selected for the present review used various measurement instruments including laser Doppler perfusion imaging (LDPI) with a PIM system  , 25 and Local reactions  induced by acupuncture manipulations are believed to be an important factor in the induction of its effects. Min et al.  studied Litscher et al.  reportedAlthough the establishment of proper control procedures in the field of acupuncture research is essential, the issues associated with acupuncture control remain difficult and the focus of contentious debate , 31. VarTaken together, these findings indicate that there are a generally higher sensitivity to pain and other abnormalities during the perception of somatosensory information in patients with pain and that these result in different responses to superficial acupuncture based on the particular conditions of the subject. Additionally, these findings suggest that, in the case of selecting acupoints close to the disease site, minimal stimuli, such as superficial needling, may be superior to deep needling in terms of increasing skin and muscle blood flow in fibromyalgia patients. Thus, the results suggest that comparisons of acupuncture with superficial acupuncture should be undertaken with caution due to the contradictory physiological effects depending on the patient's condition. It is possible that superficial acupuncture could be considered as a viable intervention as different doses or separate stimulation methods under certain circumstances.The studies selected for the present review propose several possible mechanisms that may underlie the changes in microcirculation following acupuncture treatment. Acupuncture-induced neural signaling is thought to result in alterations of blood perfusion via the local action of vasodilative factors such as substance P and CGRP , 17–19. The present review has several limitations. First, there were heterogeneities among the selected studies in terms of populations, intervention methods , control groups, and outcome measures that made reliable comparisons difficult. In particular, the assessment of outcome measures widely varied in terms of measurement devices, outcomes, measurement sites, and measurement times; thus, it was not possible to pool the data. Second, during the evaluation of the accuracy of each experiment, it was not possible to include sufficient information regarding whether each study was properly performed. Thus, the present review assessed only whether items related to the precision of the experiment were reported and, therefore, the results of this review should be interpreted with caution. Finally, there were methodological shortcomings of included studies such as the use of small sample sizes and a lack of information concerning sequence generation and allocation concealment, and these factors need to be seriously considered to design further studies.The present review determined that acupuncture induces transient reductions in blood flow due to sympathetic activation that are followed by subsequent increases in local blood flow in the skin and/or muscle. Additionally, the depth of needling and the condition of the patient may result in different microcirculatory changes, which suggests that acupuncture may be used as an additional surrogate outcome measurement in pain studies and underscores the remaining questions about whether placebo acupuncture is actually physiologically inert. Other factors to consider include the dose of acupuncture, because it may act as a confounding variable when comparing studies investigating changes in skin blood flow. The current evidence regarding the local effects of acupuncture on blood flow remains too limited to draw firm conclusions due to the small number of well-designed studies. Future well-designed studies are necessary to address these issues.Details regarding study design, participants' characteristics, measurement accuracy, and experimental environment are shown.


Tumors are not merely masses of neoplastic cells but complex tissues composed of cellular and noncellular elements. This review provides recent data on the main components of a dynamic system, such as carcinoma associated fibroblasts that change the extracellular matrix (ECM) topology, induce stemness and promote metastasis-initiating cells. Altered production and characteristics of collagen, hyaluronan and other ECM proteins induce increased matrix stiffness. Stiffness along with tumor growth-induced solid stress and increased interstitial fluid pressure contribute to tumor progression and therapy resistance. Second, the role of immune cells, cytokines and chemokines is outlined. We discuss other noncellular characteristics of the tumor microenvironment such as hypoxia and extracellular pH in relation to neoangiogenesis. Overall, full understanding of the events driving the interactions between tumor cells and their environment is of crucial importance in overcoming treatment resistance and improving patient outcome. Tumor progression is partly a result of evolving crosstalk between different cell types within the tumor and its surrounding supportive tissue or tumor stroma . InvasivCarcinoma associated fibroblasts (CAF) are believed to influence tumor behavior and outcome and thus knowledge of their biology is of importance to an overall understanding of cancer. CAFs are large, spindle-shaped mesenchymal cells that share characteristics with smooth muscle cells and fibroblasts . They coCAFs promote tumor progression in several ways such as secretion of multiple factors and MMPs, inducing stemness, EMT, epigenetic changes, etc. ,16. ReceCancer initiation and progression are largely dependent on the physical and chemical features of the adjacent environment particularly matrix topology (architecture) and stiffness. These features are determined by the size of biopolymer  fibers and the density of the fiber network . ConnectMatrix concentration and post-translational modifications such as glycosylation and cross-linking (e.g. by LOXL1 and LOXL2) affect the mechanical properties, including visco-elasticity or stiffness (reviewed in ). TumorsECM topology and stiffness can influence mechanosensing and activate intracellular signaling to promote directional cell migration. Integrin receptors and the physical arrangement of adhesions could trigger orientation of the cytoskeleton, and matrix orientation can also stabilize leading edge protrusions to promote directionally persistent migration in which specific signaling pathways  are involved ,46. CancAnother important tumor characteristic is growth-induced solid stress Figure . As tumoBesides endothelial cells and fibroblasts, the tumor microenvironment also harbors innate and adaptive immune cells. It is a complex and highly dynamic system that should concomitantly work to eradicate a tumor. However, once a system is deformed, immunity becomes a benefit for the tumor and provides a very important cue to its development and progression . The tumImmune cells, particularly macrophages and neutrophils are sources of chemokines, growth factors and proteases, as well as DNA-damaging reactive oxygen and nitrogen species. The gene expression profile of macrophages isolated from malignant tumors significantly differs from wound or resting peritoneal macrophage profile, with increased number of proliferation-associated genes . ChemokiTumor angiogenesis is an important factor in proliferation, metastasis, and drug sensitivity. Primary tumors without vasculature are small and dormant, while the growth of the tumor mass creates hypoxic conditions in the center of the tumor that induce expression of VEGF and subsequent tumor vascularization . CAFs arHypoxia is also characteristic of abnormal tumor microenvironment that is intrinsically linked to the formation of neovasculature and is clinically associated with metastasis and poor patient outcome ,84. Diffin vitro[in vivo[Low extracellular pH is another consequence of the abnormal metabolism in the tumor and supportive factor for its progression. Products of anaerobic glycolysis - lactic acid and carbonic acid (produced by carbonic anhydrase from CO2 and H2O), are the known sources of H + ions in tumors ,95. The in vitro. Coordino[in vivo indicateo[in vivo. Tissue Another feature of the pathophysiology of the tumor microenvironment, is elevated interstitial fluid pressure (IFP) ranging from 10 to 100 mmHg ,99 whileInteractions during which the tumor creates a microenvironment favourable for proliferation, for the recruitment of new blood vessels, and for the stimulation of the production of proteases that can degrade adjacent tissues, increase the likelihood of tumor development and invasion. Growing evidence points towards a key role of the multiple cellular and noncellular components of the tumor microenvironment such as cancer associated fibroblasts, immune and endothelial cells as well as matrix topology and stiffness, interstitial fluid pressure, growth induced solid stress, hypoxia and extracellular pH. Full understanding of all the events driving the interaction of tumor cells with their environment is of crucial importance in overcoming treatment resistance and in better patient outcome.The authors declare that they have no competing interests.GK conceived the idea, did literature search and drafted the manuscript; DS did literature search and contributed in drafting the manuscript; MG, NN and KB were involved in discussion and drafting the manuscript; JB supervised the project, made substantial contributions to the concept and design of manuscript, and drafted the manuscript. All authors read and approved the final manuscript.


In this article, we describe the potential benefits provided by using AFS cells in the modeling of human genetic diseases.In recent years, great interest has been devoted to the use of Induced Pluripotent Stem cells (iPS) for modeling of human genetic diseases, due to the possibility of reprogramming somatic cells of affected patients into pluripotent cells, enabling differentiation into several cell types, and allowing investigations into the molecular mechanisms of the disease. However, the protocol of iPS generation still suffers from technical limitations, showing low efficiency, being expensive and time consuming. Amniotic Fluid Stem cells (AFS) represent a potential alternative novel source of stem cells for modeling of human genetic diseases. In fact, by means of prenatal diagnosis, a number of fetuses affected by chromosomal or Mendelian diseases can be identified, and the amniotic fluid collected for genetic testing can be used, after diagnosis, for the isolation, culture and differentiation of AFS cells. This can provide a useful stem cell model for the investigation of the molecular basis of the diagnosed disease without the necessity of producing iPS, since AFS cells show some features of pluripotency and are able to differentiate in cells derived from all three germ layers “ Despite continuous increase in our knowledge about the genetic basis of a number of congenital and late-onset human diseases, so far a large majority of these conditions still remain untreatable. This is largely due to the lack of information about the precise sequence of early molecular events occurring during tissue development and underlying the pathogenesis of the disease. Indeed, although gene mutations or chromosomal aberrations responsible for the disease are present in all cells of affected individuals, the functional damage generally involves only a few tissues (or even a single one) in which the expression of the affected gene(s) occurs, affecting normal cell function. The collection of human tissues other than blood from living patients is hampered by several factors, and in most cases, at the time of diagnosis, the disruption of the cellular and/or tissue function has already occurred, making it impossible to get a clear picture of the progressive involvement of different pathways during cell differentiation. The use of animal models for the study of the consequences of gene mutations during development, although able to provide useful information, does not produce results which can be entirely translated to humans, due to the anatomical and physiological differences between the two species . Indeed,et al. (2003) [et al. described that human AF contains a fibroblast-shaped cell population positive for mesenchymal markers, such as CD90, CD105, CD73 and CD166, but negative for the hematopoietic markers, such as CD45, CD34 and CD14 [et al. (2007), who isolated c-Kit (CD117) positive populations with high clonogenic potential [In vitro experiments have demonstrated the ability of these cells to differentiate into all three germ layers giving rise to adipogenic, osteogenic, myogenic, endothelial, neural and hepatic cells, under appropriate culture conditions [Human amniotic fluid obtained during the process of amniocentesis contains a heterogeneous cell population, originating from embryonic and extra-embryonic tissues. The properties of AFS cells vary with gestational age and different approaches have been identified to isolate and characterize these types of stem cells ,18,19. B. (2003) . In the and CD14 . Thereafotential . In vitrnditions ,27,28,29nditions . Unlike nditions .+ AFS cells. Based on reports, there is scientific evidence that c-Kit+ and unselected AFS cells show similar but not identical properties and are both able to produce lineages representative of the three germ layers [in vivo in a rodent model of ischemic stroke, where transplantation of human AFS cells facilitated a reduction of the injured area, together with increment of endogenous cell proliferation and subsequent differentiation into neuronal lineage in the host brain [To date, several protocols have been used for the isolation and differentiation of AFS cells. Although the majority of studies are based on c-Kit selected cells ,32, othem layers ,36,37. Fm layers  and diffm layers ,39. In am layers ,41,42. Tst brain ,44. Of pst brain . The sec+/− selected) show common characteristics with primordial germ cells (PGC) including expression of Fragilis, Stella, Vasa, c-Kit, Rnf17, DAZAL, thus suggesting a possible origin of AFS cells from epiblast-derived cells such as PGC or PGC progenitors [In the last few years, several groups reported that both first and second trimester AFS cells  so far there is no data on their ability to produce chimeras when injected into blastocysts. Since the unique abilities of hPS consist of self-renewal, differentiation into derivatives of all three germ layers, and formation of clonal lines in vitro and teratomas in vivo, we can conclude that AFS cells are not true hPS. However, AFS cells display molecular and biological characteristics closer to hPS than to multipotent stem cells, expressing markers of pluripotency  both at the mRNA and protein level and being able to form monoclonal lines capable of differentiating in distinct lineages representative of the three germ layers. In 2010, for the first time, it was reported that clonal human c-Kit+ AFS cells have the ability to form embryoid bodies (EB) just as ES cells and PGCs [et al. demonstrated that first trimester human c-Kit+ AFS cells are able to form beating EB with high efficiency [in vitro EB with pluripotency potential and features of early stage embryogenesis [AFS cells may correspond to a new class of stem cells with properties of plasticity intermediate between embryonic and adult stem cell types in terms of their versatility ,48. In tand PGCs . EB are and PGCs  and onlyficiency . In thisogenesis . These cA comparison of the main properties and differences between ES, iPS and AFS cells is summarized in et al. showed that human 1st trimester AFS cells can be fully reprogrammed to pluripotency with non-viral methods and non-integrating systems, solving the problems related to genome integration of transgenes and the potential risk of virally induced tumorigenicity [Due to the previously described limitations of iPS cells in the modeling of human genetic diseases, AFS cells have been thought to be a viable alternative to the use of human ES and iPS. Indeed, these cells could represent a good resource for disease modeling and, consequently, one of the most promising tools in medical genetics. AFS cells can be isolated from the AF of women undergoing prenatal diagnosis to assess the presence of genomic aberrations or genetic mutations in the fetus. In the last few years, the proportion of advanced maternal age pregnancies has been rising worldwide due to changes in lifestyle; as a consequence, the number of fetuses carrying chromosomal aberrations, such as trisomy 21, is unfortunately increasing. On the other hand, the development of non-invasive prenatal diagnosis techniques, such as the analysis of cell-free DNA in maternal blood, together with the improvement of the standard first-trimester screenings, such as nuchal translucency and biochemical analysis, are allowing for a better selection for women undergoing amniocentesis allowing the isolation of an increased number of AFS cell lines carrying abnormalities. Furthermore, since AFS cells can be easily reprogrammed through a wide variety of methods ,57,58,59genicity . In thisin vitro model of Down syndrome allowing the modeling of the neurogenesis in AF-iPS with trisomy 21 and highlighting the role of miR-155 and miR-802, both encoded by chromosome 21, in the impairment of neuronal differentiation [in vitro pathogenic phenotypes, and the establishment and banking of clonal AFS cell lines derived from pregnancies with specific genetic aberrations would be of highest value for basic research and drug discovery [in vitro. Preliminary results of our group suggest that AFS cells are capable of generating human cellular disease models in vitro by CRISPR/Cas9 system with great speed and efficiency (data not shown).A few examples of this possibility have already been reported in the literature. Fan and colleagues, in 2012 , generatntiation ,60. Moreiscovery   [in vivo-like properties that make them suitable to be used in both targeted and phenotypic/physiological pharmacological screening as well as in drug neurotoxicity evaluation [9tetrahydrocannabinol which accounts for the majority of the reproductive hazards in marijuana preparations [Drug development is a costly and long multi-phase process burdened with a high rate of failure mainly occurring in late phase III trials or at registration. This happens in a scenario in which both libraries containing thousands of testable compounds with the potentiality to become therapeutic agents and technological platforms for high-throughput screening are dramatically improved. One recognized reason for the high rate of failure of many drug candidates is the poor predictability of the preclinical studies carried out on animal models, which are mined by significant species-specific differences in the genetic background and in the mechanisms underlying the disease etio-pathology. The use of tocytes) . The advtocytes) , allowintocytes) . The modtocytes) , isoprottocytes) , IGF1 antocytes) . Moreovetocytes) . Increasaluation . Could Aaluation ,68, whicarations .In relation to their utility for evaluating drug cardiotoxicity and hepatotoxicity, studies have also demonstrated the differentiation potential of AFS cells into cardiomyocytes and hepatocytes ,71,72,73On the other hand, although AFS cells have been reported to promote nerve regeneration and myelination and to produce several cytokines and neuro-glial factors , there iAFS cells have been also reported to form three-dimensional (3D) chimeric organoids with mouse embryonic kidney cells . The humIn conclusion, AFS cells, with their potential large-scale amplification, differentiation and 3D organoid formation, could represent a very interesting source to produce physiologically-relevant novel systems to adequately test pharmaceutical agents before their administration to patients, although additional experimental data that confirms this hypothesis is still required .in vivo studies are supporting the observations previously reported in the epidemiological studies and have begun to attribute the transmission of various traits to changes in sperm [in vivo [in vitro model mimicking human gamete formation would be an extremely valuable research tool. As previously described, AFS cells have been recently demonstrated to share several features with PGC, thus suggesting their use as an efficient handy tool to fulfill the scarcity of in vitro models to study human gametogenesis and fill the void of insufficient data on germ-line diseases and diseases caused by epigenetic alterations endangering trans-generational inheritance.While an organism’s genotype is static throughout its life, the epi-genome is highly dynamic and can adapt or be altered in response to the internal or external environmental factors. Germ cell development is a critical period during which epigenetic patterns are established and maintained. If epigenetic errors are introduced at any time during germ line development, some serious consequences for fertility and the health of future offspring(s) may arise . The effin sperm . Most rein sperm  sparked in sperm . Furtherin sperm . Despite[in vivo . Therefo(1)AFS cells can be easily collected from women undergoing amniocentesis, a prenatal diagnostic tool currently used in Western countries in a growing number of pregnancies, mostly due to the increased cases characterized by advanced maternal age caused by social and cultural factors. The recent improvements in the first trimester non-invasive tests for prenatal diagnosis will likely reduce the total number of amniocentesis, but will increase the rate of genetically abnormal pregnancies, thus providing a large number of samples to be used for the modeling of specific genetic diseases;(2)Unlike iPS, AFS cells will provide the opportunity to also investigate diseases that are lethal during pregnancy. Moreover, AFS cells are less prone to aging dependent genetic and epigenetic modifications as compared to fibroblasts, commonly used for the generation of iPS, thus providing a more natural model for the study of the disease;(3)AFS cells can be easily reprogrammed by using both viral and non-viral methods, making this process very efficient. Moreover, the use of non-viral methods will likely reduce the cost of the reprogramming procedure as compared to the use of iPS, which also requires the availability of specific facilities for the use of viral vectors.Based on literature data, AFS cells have been so far proposed to be an interesting model for cell therapy, due to their ability to differentiate into different cell types and their lack of ethical concerns. It is important to underline that disease modeling represents an appropriate alternative to animal experimentation because it allows overcoming all limitations related to preclinical research such as the ethical implications and excessive costs of animal models. In light of data described in the present review, it can also be hypothesized that AFS cells will become one of the main models for the modeling of human genetic diseases in the near future. The advantages of this model as compared to the iPS cell paradigm can be summarized as follows:in vitro models, allowing the procurement of AFS clones with induced mutations and, consequently, the study of a wide range of pathogenic conditions. The recent advances of functional genetics including genome-scale loss-of-function drug-suppressor screens, obtained by the application of the CRISPR-Cas9 technique, can be successfully applied to these kinds of cells, further creating novel opportunities for testing the modes of action and the efficacy of various drug candidates.It can also be hypothesized that, in the coming years, the newly discovered genome editing technique, CRISPR, could provide a useful tool to increase the number of


One thesis summary has been published in the current issue of IJIC [In recent decades, the number of published health economic studies – especially economic evaluation studies – in the field of  of IJIC . In geneSeveral authors have discussed these issues, highlighting the fact that there are still many scientific challenges. As such the field of ‘health economics of integrated care’ is still in its infancy stage –6. The kTo discuss these problems, challenges and the growing relationship between health economics and integrated care, a meeting was held during the congress of the International Foundation for Integrated Care (IFIC). The meeting took place in Brussels in 2014, with about 20 participants. Four challenging areas were identified and discussed: (1) searching literature and assessing the quality of the integrated care literature, (2) outcome measurement and valuation in integrated care, (3) determination of the costs of integrated care and (4) several other design and analysis aspects.The first area of challenges is related to the search for and assessment of the quality of literature on integrated care. This subject is especially important for model-based economic evaluation studies. For modelling, these studies rely on a systematic synthesis of existing literature. This requires well-defined and clear strategies to search for the right literature and the presence of evidence-based checklists to determine the methodological quality thereof. With respect to integrated care, sensitive and specific search strategies as well as a quality assessment checklist are still lacking.The second challenging area is the outcome measurement and valuation of integrated care. A proper measurement and valuation of integrated care requires the incorporation of its main characteristics. Among others, they include the fact that integrated care often has a multi-outcome focus instead of a focus on a single  outcome. Integrated care is often aimed at improving client satisfaction, access, affordability, sustainability and (thus) at improving the process of care. Moreover, integrated care arrangements do not only focus on the individual but also on the health and well-being of significant others. Next to this, the relevant domain of the outcome measurement for the client can vary when going through the integrated care chain. Clients, for instance, might value outcomes in the acute phase differently than in the after-care phase. Traditionally, in health economics, single  outcomes, quality of life or quality adjusted life years for single patients  are used as an important measure to express outcomes. Given the specific characteristics of integrated care, a considerable methodological challenge in outcome measurement is thus to develop and/or establish sensitive generic outcome valuation methods aiming at providing accurate and relevant information on the outcomes of integrated care throughout the whole care chain, for both the client as well as his/her significant others.The third range of challenges is related to the determination of the costs of integrated care. To properly calculate these costs, a cost method is required which is robust enough to incorporate particular features. These include, among others, continuous changes in the delivery of integrated care (so a heterogeneous production process), the absence of standardised outcomes (so no clear outputs) and the presence of many indirect costs (because of many coordinating activities and cooperative actions). Traditional cost methods are usually output-oriented and assume homogeneous production patterns, uniform standard outputs and the importance of direct costs only . Given tFinally, during the Brussels meeting, other design and analysis aspects were discussed. These relate to an adequate study design, such as a naturalistic study design instead of the randomised controlled trial, definition of comparators, dealing with missing data, sound statistical analysis and standardised guidelines for reporting scientific research on health economic approaches of integrated care.KST@euro.who.int).In summary, during the last decades the methodological challenges related to health economics and integrated care have been highlighted by several experts –7. Now t


In vitro ROS levels, the mRNA level and activity of superoxide dismutase, glutathione peroxidase and catalase were obtained. The results showed that in comparison with negative control, ZEA increased oxidative stress with higher ROS levels, reduced the expression and activity of antioxidative enzymes, increased the intensity of fluorogenic probes 2’, 7’-Dichlorodihydrofluorescin diacetate and dihydroethidium in flow cytometry assay and fluorescence microscopy. Meanwhile, the activity of glutathione (GSH) did not change obviously following 60 μM ZEA treatment. Furthermore, the underlying protective mechanisms of curcumin on the ZEA-treated porcine granulosa cells were investigated. The data revealed that curcumin pre-treatment significantly suppressed ZEA-induced oxidative stress. Collectively, porcine granulosa cells were sensitive to ZEA, which may induce oxidative stress. The findings from this study clearly demonstrate that curcumin is effective to reduce the dysregulation of cellular redox balance on porcine granulosa cells in vitro and should be further investigated for its protective role against ZEA in animals.Oxidative stress (OS), as a signal of aberrant intracellular mechanisms, plays key roles in maintaining homeostasis for organisms. The occurrence of OS due to the disorder of normal cellular redox balance indicates the overproduction of reactive oxygen species (ROS) and/or deficiency of antioxidants. Once the balance is broken down, repression of oxidative stress is one of the most effective ways to alleviate it. Ongoing studies provide remarkable evidence that oxidative stress is involved in reproductive toxicity induced by various stimuli, such as environmental toxicants and food toxicity. Zearalenone (ZEA), as a toxic compound existing in contaminated food products, is found to induce mycotoxicosis that has a significant impact on the reproduction of domestic animals, especially pigs. However, there is no information about how ROS and oxidative stress is involved in the influence of ZEA on porcine granulosa cells, or whether the stress can be rescued by curcumin. In this study, ZEA-induced effect on porcine granulosa cells was investigated at low concentrations .  Fusarium and is found to cause mycotoxicosis in animals and humans , 532-) Granulosa cells as a primary unit of ovarian follicle are critical to guarantee the maturation of oocyte and maintain the normal level of hormones in animal. Due to the hormone disruption by ZEA, the proliferation of porcine granulosa cells decreases while as the ratio of apoptosis enhances at high concentrations of ZEA . The gro


A spatially continuous mathematical model of transport processes, anaerobic digestion and microbial complexity as would be expected in the human colon is presented. The model is a system of first-order partial differential equations with context determined number of dependent variables, and stiff, non-linear source terms. Numerical simulation of the model is used to elucidate information about the colon-microbiota complex. It is found that the composition of materials on outflow of the model does not well-describe the composition of material in other model locations, and inferences using outflow data varies according to model reactor representation. Additionally, increased microbial complexity allows the total microbial community to withstand major system perturbations in diet and community structure. However, distribution of strains and functional groups within the microbial community can be modified depending on perturbation length and microbial kinetic parameters. Preliminary model extensions and potential investigative opportunities using the computational model are discussed. The gastro-intestinal tract (GIT) describes the collection of organs involved in converting ingested materials into usable energy sources while removing additional waste products. Digestion predominantly occurs in the upper GIT  while final nutrient extraction and fecal preparation (water absorption) occurs in the lower GIT . Energy in vivo and in vitro, have been developed and used extensively  of all materials considered in the colon-complex. We describe functions R, F and E in detail in the following sections, but present By way of material balance, we can combine the three sub-processes and describe the density of materials in the colon with the following advection-reaction system:Physiological systems are highly complex, functioning with redundancy, time-variations and interplay with other systems . Rather Colon Geometry: A cross-sectional slice of the colon would display highly irregular geometry, as there exists mucousal folds and villous surfaces [surfaces , 27. ForMaterial Properties: To be consistent with our first assumption (1D tube geometry), we assume that the materials in localized portions of the colon  will be homogeneous (well-mixed) across the cross-sectional area. Additionally, we only measure water generated through the fermentation process, rather than try to account for all water balances through interaction with other physiological processes.Mucus Thickness: Mucus is produced endogenously throughout the colon. The rate of this mucus production is constant along the main flow direction and across the mucus thickness. Additionally, we treat this layer as a fixed medium of constant volume, with the volume of the mucus being 10% of the total colon volume.Transit Time: The effect of peristalsis and additional propulsion mechanisms manifest as an average flow or speed of convective transport. This allows us to approximate convective transport as a first-order flux with constant velocity term.Metabolic Pathways: The only macromolecules reaching the colon are carbohydrates, and the anaerobic digestion of carbohydrates follows the metabolic pathway described in [ribed in . As a stribed in , 3, we aribed in , 21, 28.Reaction Processes: Combining the processes involved in metabolism and the natural decay of flora in the system, we can summarize the reaction processes in the flora-diet system as: (1) hydrolysis, (2) glucose utilization, (3) lactate utilization, (4) acetogenesis, (5) methanogenesis, (6) decay of SD flora, (7) decay of LD flora, (8) decay of HDA flora, and (9) decay of HDM flora. The choice of metabolic pathways and subsequent reaction processes is responsible for the overall model problem size, thus a simpler representation of anaerobic digestion would lead to a smaller state-space, and a more involved representation of anaerobic digestion would lead to a larger state space.We remark that the model assumptions and simplifications can be relaxed on future model iterations as knowledge of functional details continues to grow, but doing so would require the inclusion of additional mathematical and numerical complexities. Complete mathematical formulation and details are provided in blackbox approach to describe the upper GIT as a system of dilution-units is employed. This approach effectively assumes there is continuous flow through and limited biochemical changes to material in the upper GIT.The choice of sub-models, particularly of anaerobic digestion/metabolic pathway, is key to determining the size and structure of the overall model. We build upon the model of  which dehttp://compugut.sourceforge.net).The developed mathematical model of context driven problem-size and functionally defined sub-processes presents significant organizational challenges during numerical simulation. Additionally, simulation of large models will invariably create large data-sets, both with analytical and visualization challenges. The compuGUT software project stems from these design challenges, providing interested users a preliminary model implementation for review and experimentation. The compuGUT suite of tools was developed for Linux systems, and performs three primary tasks: (1) Simulation Design, (2) Numerical Integration, and (3) Data Visualization. Tasks (1) and (3) are performed by auxiliary tools written as either BASH or R scripts, while task (2) is performed by our primary compuGUT tool written in C, which is a flexible implementation of a standard central-flux scheme for balance laws . The comeMT carbohydrate digestion model, we execute a series of simulation experiments/scenarios using the compuGUT. Source codes and analytical scripts used to perform and study the following simulation scenarios are available on the compuGUT project page.To study the features of the spatial continuity and As noted, the model of  assumes D and %BiomassC are the percent biomass values generated at all locations and time steps using the 3-stage discrete physical model and the spatially continuous physical model, respectively, and percent biomass is tabulated as:To analyze the performance of both reactor representations, we extrapolate the solution of the three-stage reactor over the length of the entire colon, and then compare the difference in their prediction of material composition, particularly percentage of total material that is a biomass quantity, as a percent deviation:By extending the description of carbohydrate digestion of  to incluTo assess the value of added microbial complexity beyond parameter sensitivity, we simulate hypothetical system scenarios with differing microbial representations. Particularly, we look at how the entire colon-complex responds to a sudden/temporary drop off in dietary fiber consumption, or to a sudden/temporary removal of the major or predominant strains/representatives within the sugar utilizing biomass functional group.For diet perturbations, we execute a 10 week simulation with diet profile as outlined in To analyze diet perturbations, we compare the average percent biomass  produced in each of the 4 microbiota representations  for all locations along the length of the colon and at all times during the entire simulation.Microbial communities can be disturbed due to a variety of physiological and/or environmental conditions. We define microbial community perturbation in our simulations as an an increased cell lysis rate of the dominant representative strain in the sugar utilizing biomass functional group. An alternative approach to community perturbation could be increased removal rates (washout) of a dominant strain, but given the current stage of our flow model development, increasing cell lysis rate was considered a convenient means to perturb the microbial system in a easily quantifiable way. We simulate systems of 5 sugar degraders, and perturb each system for a fixed number of days (between 0 and 10). To analyze the output of these simulations, we first look at the percent biomass (as described previously) at the output of the colon to assess the macroscopic behavior of the system, and then look at the detailed community structure by assessing the percent composition of each of the 5 sugar utilizing biomass representatives. Simulation scenarios were run with a biochemical deviation of 1% and are repeated 6 times to evaluate the effects of stochastically chosen parameters.A single 8 week simulation of the compuGUT with single representatives within biomass functional groups (MT model), normal physiological conditions , and coarse-most numerical discretization generates 2821 text files with information regarding the concentration of all state-variables at all locations. It should be noted that the anaerobic digestion model used for simulation only includes gas components in dissolved form rather than both dissolved and vapor. This results in concentrations of 28 dependent variables at the minimum 51 colon locations. A visualization of coarse-composition, which we define as material categorized by type as either fiber, biomass, sugar, short-chain fatty acids (SFCAs) or others, is shown as In assessing the coarse composition of material along the length of the colon using both the three-stage physical representation and the spatially continuous representation, we find substantial differences between both model outputs. Microbial representation has significant effect on coarse colon description during simulations with dietary perturbations . Simulathealthy, state within 4 days. Increased microbial representation allows for the colon process model to recover performance during major disruptions to the microbiota community. in vivo and in vitro. With future modifications to the compuGUT output measurements, we may have the opportunity to generate results quantitatively comparable to those generated through experiment. With newly developing spatial measurement technology to measure aspects of digestion through the length of the system in live animals, such as the Smart Pill [Significant developments regarding microbiota and human interaction have been made with respect to experimental measures and analytical techniques , creatinart Pill , 34, dirin vitro experimental systems, as well as previous mathematical models of carbohydrate digestion in the colon [One of the primary simplifications made during model development is the physical representation underlying the model structure—a spatially continuous pipe with fixed-media attached to pipe walls and fluid media convected in a single direction. Traditional he colon , employ he colon  and confin vitro and in silico models, it cannot guarantee complete description of colon physiology. Most notably, our physical representation creates a gross simplification of the exchange processes . Similarly, we employ a fairly substantial simplification regarding material properties and flow dynamics, which also creates most difficultly in assessing exchange processes. Given our current goals of modeling microbiota behavior in the colon and the top-down structure of our model leading to short-chain fatty acid concentrations not affecting microbial densities, the accuracy with which we model exchange processes is not paramount. That said, short-chain fatty acids produced in the colon are of great importance to the physiological function of the colon [The physiological colon is spatially continuous, but of irregular cross-sectional geometry due to villi and compression . Though he colon –37, and chyme in a model of digestion in the small intestine. The challenge of implementing such a description in our current framework is the presence of intestinal microbiota. In the small intestine, digestion is a chemical process independent of water availability. In the large intestine, digestion is completed by anaerobes and so the rate of digestion is highly dependent on water availability, strengthening the mathematical coupling of digestion, flow, and exchange processes. Thus incorporating viscosity changes may require an overhaul of how reaction and exchange parameters are defined, but such an overhaul will provide a model much more capable of describing behaviour closer to the true colon.An approach to better capture physiological accuracy is to refine the convective flux model to segregate peristaltic forces rather than average these forces to a singular expression, and to capture the changing material properties of the digesta passing through the colon. In , 39 the steady state. Potential health implications of gut flora diversity changes are discussed in [As noted, we extend the MT-model of carbohydrate digestion by anaerobic microbiota in  using anussed in , 41. Basussed in . Though coarse material composition, as well as microbial community structure, but rigorous mining and analysis of simulation data could lead to additional insights that we have overlooked.At its present state, the computational model is limited to modifications of input dietary fiber, initial conditions, physical parameters  and microbial representations. However, we have yet to fully explore the complete model output space due to its high dimensionality. That is, the model is capable of generating a vast amount of spatial and dynamics data that can be interpreted in a variety of lights. In this paper, we presented perspectives using With model extensions to better describe material/flow properties incorporating the concept of viscosity and muscular peristalsis, and a more complete digestion process including digestion of multiple substrates, i.e. the model of , as wellin vitro and in vivo experimentation should be conducted.We presented a spatially continuous model of carbohydrate digestion and transport processes in the colon—an extension of the three-stage model described in . Both moThe extended microbial representation in the description of carbohydrate degradation by intestinal anaerobes provides us the ability to simulate system perturbations. We find that the overall system performance, judging how the anaerobic digestion process proceeds, during a period of distress is tempered by having a diverse microbial community present. However, the composition of the microbial community after a distress/perturbation period is often not the same as it was prior to that period. How the post-distress microbial community affects host-health is an important follow up question.S1 AppendixThis document details the construction of the underlying mathematical models, and includes important information necessary for the reproduction of the numerical program.(PDF)Click here for additional data file.


Incidence of most common cancers increases with age due to accumulation of damage to cells and tissues. Stroma, the structure close to the basement membrane, is gaining increased attention from clinicians and researchers due to its increasingly, yet incompletely understood role in the development of age-related cancer. With advanced age, stroma generates a pro-tumorigenic microenvironment, exemplified by the senescence-associated secretory phenotype (SASP). Components of the SASP, such as cytokines, chemokines, and high energy metabolites are main drivers of age-related cancer initiation and sustain its progression. Our purpose is to provide insight into the mechanistic role of the stroma, with particular emphasis on stromal fibroblasts, on the development of age-related tumors. We also present evidence of the potential of the stroma as target for tumor therapy. Likewise, a rationale for age-related antitumor therapy targeting the stroma is presented. We expect to foster debate on the underlining basis of age-related cancer pathobiology. We also would like to promote discussion on novel stroma-based anticancer therapeutic strategies tailored to treat the elderly. Aging is a complicated process associated with accumulation of damage to cells and tissues resulting in attenuated or dysregulated function and increased risk of disease including cancer . MechaniThe purpose of this minireview is to provide insight into the mechanistic role of the stroma in age-related cancer. Our focus is on stromal fibroblasts, because of growing evidence suggesting their role in many aspects of tumorigenesis. Understanding the involvement of components of the stroma in the regulation of age-related diseases, particularly cancer, may help to increase our understanding of the basis underlining pathobiology of age. Gained knowledge may also help us to envision novel therapeutic strategies specially tailored to treat the elderly.Stroma is a collagen-rich support structure close to the basement membrane where the tissue resides. The main components of stroma and basement membrane are produced by stromal fibroblasts ; their aIn organs, stroma and parenchyma provide supportive framework and key elements essential to normal functioning. Resident cells and structural factors stably occupy the stroma  and evenAmong non-resident constituents of tumor stroma, immune cells represent a substantial percentage . They inAs a result of replicative exhaustion, normal diploid differentiated cells become permanently arrested on the G1/G0 cell-cycle phase . AlthougFor the purposes of our succinct revision, we focus our discussion in senescent fibroblasts as contributors of age-related tumorigenesis. Mounting studies sustain our case. It is, however, necessary to note that the quest for establishing the putative role of senescent fibroblasts in promoting age-related tumorigenesis in humans is a very active topic of scientific discussion and more work is needed to settle the debate. It is also necessary to note that the effects of advanced age on other components of the stroma, not analyzed in detail in this revision, must be considered when discussing the complex microenvironmental interactions driving age-related tumorigenesis. For instance, immunosenescence affects adaptive and innate immune cells . Along win vitro transformation of fibroblast cell lines , which subsequently acts upon the stromal fibroblasts and renders them senescent . This evAlthough cellular senescence is a mechanism of aging, the fact that senescent cells do not divide does not protect the elderly against tumorigenesis. On the contrary, age-associated microenvironmental alterations seem to be the main driver of tumor development . AccordiDespite recent advances presented herein, the mechanistic role of stromal cells in modulation of age-related cancer is still controversial reflecting the limited studies in this area to date. Although considerable progression has been made in recent years, there are still numerous key questions that need to be addressed. Some of these questions pertain to the regulation of stromal senescence, microenvironmental changes that initiate age-associated stromal senescence and outgrowth of cancer, and the mechanistic role of senescent stromal fibroblasts in the modulation of cancer initiation and progression in the elderly. Whether senescence of specific components of the stroma is sufficient by itself to initiate carcinogenesis still remains to be determined. However, the ability of senescent fibroblasts to create a tumor microenvironment by their SASP may serve as a model to explain in part how increased tumor incidence is observed in aged individuals.+ T cells engineered to deliver IL-12 within tumor stroma triggered an acute inflammatory environment, improved antigen presentation by myeloid-derived cells within tumors, increased infiltration of adoptively transferred antigen-specific CD8+ T cells, and eventually induced regression of an established murine melanoma . In turn, cancer cells import these compounds via the MCT1 transporter (N-acetyl-cysteine (NAC) increases the lifespan in different experimental models -derived ROS  and prostate cancer (PCa) . Growingived ROS . Similarived ROS . Despiteived ROS . These rived ROS . Howeverived ROS . Moreoveived ROS , suggestCaloric restriction (CR) reported to increase the lifespan in different models lowers the risk of various age-related diseases including cancer . SimilarExposure to low concentrations of dietary flavonoids and polyphenols is known to modulate the lifespan in different experimental models, by actions that are independent of their antioxidant properties . The “MeAmong the mechanisms underlying the molecular and/or cellular basis of age-related cancer, stroma stands up as a very critical player. Components of the senescent stroma, such as fibroblasts, contribute to create a tumor microenvironment through mechanisms including the contribution of the SASP, and resident and non-resident stromal component cell types. Our growing knowledge of the common chronic effects of stroma-derived factors on the promotion of aging and cancer has resulted in the characterization of molecular pathways in the stroma driving age-related cancer. Since these molecules have been explored as targets for tumor therapy, we may anticipate potential anticancer therapeutic strategies targeting the aging stroma. Yet interesting advance has been achieved, we need to know more about critical points of inhibition, disruption, or activation in metabolic pathways operating on the aged stroma. Specifically, the potential use of dietary supplementation with antioxidants, use of CR mimetics, or secoiridoid polyphenols, all of them with proven anti-tumor effects targeting metabolic components of the stroma, should be further explored. Identification of cancer and age-associated mutations in genes controlling the complex mechanisms governing the interaction between stromal components in normal aging is needed. Gained knowledge will increase our understanding of the mechanistic role of the stroma in the development of age-related cancer and will provide novel treatments for cancer in the elderly.AE drafted the manuscript, edited, and approved the final version; MH contributed to conception and design, wrote the manuscript, edited, and approved the final version; CRG contributed to conception and design, wrote the manuscript, edited, and approved final version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Neurological disorders are prevalent worldwide. Cerebrovascular diseases (CVDs), which account for 55% of all neurological diseases, are the leading cause of permanent disability, cognitive and motor disorders and dementia. Stroke affects the function and structure of blood-brain barrier, the loss of cerebral blood flow regulation, oxidative stress, inflammation and the loss of neural connections. Currently, no gold standard treatments are available outside the acute therapeutic window to improve outcome in stroke patients. Some promising candidate targets have been identified for the improvement of long-term recovery after stroke, such as Rho GTPases, cell adhesion proteins, kinases, and phosphatases. Previous studies by our lab indicated that Rho GTPases (Rac and RhoA) are involved in both tissue damage and survival, as these proteins are essential for the morphology and movement of neurons, astrocytes and endothelial cells, thus playing a critical role in the balance between cell survival and death. Treatment with a pharmacological inhibitor of RhoA/ROCK blocks the activation of the neurodegeneration cascade. In addition, Rac and synaptic adhesion proteins (p120 catenin and N-catenin) play critical roles in protection against cerebral infarction and in recovery by supporting the neurovascular unit and cytoskeletal remodeling activity to maintain the integrity of the brain parenchyma. Interestingly, neuroprotective agents, such as atorvastatin, and CDK5 silencing after cerebral ischemia and in a glutamate-induced excitotoxicity model may act on the same cellular effectors to recover neurovascular unit integrity. Therefore, future efforts must focus on individually targeting the structural and functional roles of each effector of neurovascular unit and the interactions in neural and non-neural cells in the post-ischemic brain and address how to promote the recovery or prevent the loss of homeostasis in the short, medium and long term. Neurological disorders are highly prevalent around the globe. In 2008, neurodegenerative disorders were responsible for 1% of disabilities worldwide W.H.O., . StrokesThe NVU acts as a guardian of cerebral homeostasis. Neurons, glia, the perivascular space, and the endothelium are closely interrelated to maintain the homeostasis of the brain microenvironment , this disease can result in several clinical manifestations, including paralysis or hemiplegia, aphasia, and memory and learning impairment, among others -6 and IL-1 via nuclear factor-κB (NF-κB) activation , BBB dysfunction has been detected not only as a late event but also at early stages of disease progression. There are two types of BBB dysfunction involving increased permeability: passive diffusion with edema formation and massive cellular infiltration across the BBB ) in glioblastoma cells. These findings suggest that CDK5 may play a significant role in the regulation of EC proliferation  in a hypoxia model preserves cell motility and temporal control of actin cytoskeletal dynamics and protects and promotes angiogenesis  during an action potential in presynaptic neurons. K+ passes into the blood vessel via gap junctions for removal after a long period of neuronal activity , preventing the neurodegeneration and neuronal loss in AD mice (Piedrahita et al., The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Accordingly, this approach aimed to guarantee the suitability of the final product is classified in primary chains involving the initial stages of production from breeding, nursing, weaning and growth of animals in livestock farms to the slaughter process. Secondary chains include all the subsequent stages of the primary chain, where the animal by-product is transformed, pitted, chopped, processed, stored, transported and retailed to the final consumer.2The agricultural value chain concept refers to the interaction between multiple manufacturing stages involved in the production process of a specific nourishment. The earned value of the final product is generated due to the operational or transactional costs through agro-food linear diagram and there is a livestock commodities dynamics pattern to meet expectations of consumers.3 Under the approach ''from farm to fork'' the OIE promotes harmonization of sanitary regulations for trade of animals and their products as well as the management of sanitary risks where veterinary professionals play a pivotal role. Thus, throughout the productive and value chain puzzle, veterinarians play key role in: 1) the application of risk analysis procedures,4 2) quickly and efficiently detecting diseases listed by the OIE, including those that lead to a significant risk in terms of food safety and public health; and 3) international trade of livestock inputs, as veterinary certification is a crucial requirement to export animal products. Thereby, there is a mutualistic and synergistic partnership between the agri-food production chain structure and public health veterinarian, where the functionality and competitiveness of the agricultural value chain structure to access the global markets, is a primary responsibility of veterinarians. Whereas, the interaction and partnership of each stake-holder across the horizontal chain structure ensures the integration, strengthened and correct implementation of public health policies at public and private level carried out by the veterinary services. For example, by evaluating the microbiological risks and estimating the appropriate level of protection (ALOP), food safety objectives (FSO) and performance objectives (PO); or enforcing and developing traceability programs for livestock or animal by-products.5The agreement on sanitary and phytosanitary measures (SPS agreement) of the world trade organization (WTO) recognizes the World Organization for Animal Health (OIE) standards as the reference codes for control and prevention of trans-boundary animal diseases and zoonoses, including food-borne zoonoses.6 Likewise, one of the most significant impacts of globalization is the need to upgrade veterinary services to promote and protect human and animal health and adjust local nourishment production to international trade models under the SPS Agreement standards. Therefore, agricultural value chains constitute an alternative to enhance the utilization of resources, afford compliance with environmental protocols, improve sanitary management of trans-boundary animal diseases and food-borne zoonoses, increase access to global markets, strengthen rural development paths, and establish an integral framework for the exchange of tools to reduce poverty, food insecurity and social inequality.Moreover, agro-terrorism and bioterrorism prevention, changes in socio-demographic trends in developing countries, dietary and agricultural exporter patterns promoted by globalization of livestock markets, will evolve with a consistent and progressive parallelism towards more robust and strong food security surveillance schemes.7 Hence, it is essential to strengthen both academic and professional training programs at undergraduate and postgraduate level of veterinary public health and food safety programs,8 where particular importance should be granted to manufacturing and value chain structures, as holistic and trans-disciplinary systems that contribute in a practical and tangible way to safeguard and improve the competitiveness of our local agri-food markets.In this scenario, veterinary professionals serve as catalysts to bolster rural economies by enabling the development of a sustainable livestock activity and guarantee food hygiene.


Angioedema is a highly heterogeneous group of conditions and is characterized by sudden, pronounced swelling of the lower dermis and subcutaneous. Because of its frequent coexistence with urticaria, it is often classified in the same manner as urticaria. However, it also includes categories not associated with urticaria. Angioedema without urticaria is characterized by hereditary and acquired angioedema and histaminergic and nonhistaminergic angioedema. The prevalence of subtypes of angioedema without urticaria was estimated at the Clinical Immunology outpatient service of an University Hospital, in Rio de Janeiro.We have classified 118  outpatients with angioedema without urticaria in categories: hereditary angioedema (HAE), angiotensin-converting enzyme (ACE) inhibitor-induced angioedema and idiopathic angioedema.HAE was showed in 98 of 118 patients (83%). ACE inhibitor induced-angioedema was showed in 16 patients  and 4 patients  were diagnosed with idiopathic angioedema, after a complete investigation of all causes of angioedema.In our casuistic, HAE was the most prevalent type of angioedema without urticaria, as described in the literature. However, the prevalence of ACE inhibitor-induced angioedema found in our study was higher, since the reported incidence of this condition ranges from 0,1 to 6%. Correct diagnosis of the subtypes of angioedema without urticaria should be the basis of better understanding and the treatment of these conditions.


CART) has been the subject of significant interest for over a decade. Work to decipher the detailed mechanism of CART function has been hampered by the lack of specific pharmacological tools like antagonists and the absence of a specific CART receptor(s). However, extensive research has been devoted to elucidate the role of the CART peptide and it is now evident that CART is a key neurotransmitter and hormone involved in the regulation of diverse biological processes, including food intake, maintenance of body weight, reward and addiction, stress response, psychostimulant effects and endocrine functions (Rogge et al., The cocaine- and amphetamine-regulated transcript ( Experiments conducted with acute administration of cocaine or amphetamine in rodents resulted in the upregulation of a particular mRNA species in the striatum of the brain that was subsequently named “cocaine- and amphetamine-regulated transcript” (CART) and the encoded peptides are referred to as CART peptides  in brown adipose tissue, relating CART peptides not only to the control of feeding but also the modulation of energy expenditure  has not been identified to date, there is strong evidence that CART signaling can be blocked by pertussis toxin (PTX), indicative of the involvement of an inhibitory G-protein-coupling receptor that couples to GCART mRNA-containing neurons are present in high densities in diverse regions such as the Edinger-Westphal nucleus, ganglion cells in the retina, mitral and tufted cells of the olfactory bulb, sensory barrels in the cerebral cortex, the pituitary, lamina X of the spinal cord, medulla of the adrenal cortex, the vagal nuclei, and a number of hypothalamic nuclei  /AgRP neurons which promote food intake and reduce energy expenditure, and the adjacent anorexigenic POMC/CART neurons that inhibit food intake and increase energy expenditure -mediated effects  of the striatum, whilst refeeding restored the expression  function, it is at present unclear the definite roles of enteric CART in intestinal motility, satiety and feeding behavior -1 receptors has been detected in the CCK-1 receptor mRNA expression was detected in the nodose ganglion  and the absence of specific antagonists. Nevertheless, numerous studies incorporating both pharmacological and genetic manipulations of CART expression in murine models have been endeavored in the last decades to determine the sites of action and the effects on feeding behavior and metabolism of the peptide. Overexpression studies to discern the brain regions mediating CART-induced regulation of feeding have been the most common approach in rodents. Widely adopted as an appetite-regulating peptide of the CNS with hypothalamic expression levels modulated by nutritional status   treatment in starved mice, further denoting the potential role of Acb CART in mediating the appetite suppressant properties in models of anorexia nervosa  model, which was generated from a conditional version that was crossed with an oozyte-specific Cre line. Collective evidence regarding the role of CART in energy homeostasis from a few representative studies conducted by independent research groups  mice  revealed a pronounced increase in adiposity independent of diet. The gain in whole body fat mass was further confirmed by tissue dissection showing significantly elevated fat masses in all white adipose tissues, including the inguinal, epididymal, mesenteric and retroperitoneal regions. This also suggests an important function of CART in lipid metabolism, where CART has been linked to inhibition of lipogenesis and stimulation of lipid substrate mobilization and utilization  between CART knockout models generally support the property of CART as a satiety factor and an anorexigenic signal in the brain, as evident in the elevation in body weight gain attributable mostly to the increased fat mass consistent across studies, although controversy exists for the corresponding food intake data. As for the aspect of energy intake, RER was demonstrated to be reduced both under the conditions of CART overexpression and ablation, suggesting fat was metabolized as the primary fuel for energy supply. A possible reason could be that although CART may intrinsically promote the utilization of fat as the predominant fuel source for reducing energy intake, the effectiveness of CART deletion on the disturbance of lipid metabolism hence accumulation of adiposity may have surmounted the simultaneous CART deficiency-induced enhancement of energy intake, resulting in a net reduction in RER based on the readily available fat depots.Taken together, results from the literature as well as in-house studies of CART gene is encoded  to obesity  and specific antagonists, continues to constitute the major challenge in understanding the underlying mechanisms by which CART exerts effects on feeding and neuroendocrine regulation.The physiological importance of CART is endorsed by the evolutionarily conserved expression patterns in the brain as well as the periphery at regions associated with energy balance, inferring structural hence functional conservation in glucose sensing, lipogenesis regulation and ultimately the control of feeding behavior. The resultant integrated outcome on metabolic modulation also represents the neuronal crosstalk involving similar neuromodulatory peptides between central CART-interfered pathways residing at the hypothalamic and brainstem neurons as well as via the hypothalamic-pituitary-adrenal axis. In comparison with CART in the brain, functional roles of peripheral CART are less established, with indeterminate modes of action speculated to entail either a local independent response, or synergistic effects with central CART.CART siRNA respectively reflect a concerted response from the CART-dopaminergic system interaction within the Acb, hence the observed feeding modulation and locomotive effects were likely perplexed by the inherent reward and motivation pathway. Several hindbrain areas are of particular interest as alternative brain regions for CART-induced anorexic effects, owing to the well-described role of the hindbrain in conveying post-prandial satiety effects to the hypothalamus, consonant with the indicated relation between CART and vagally-mediated gastrointestinal satiety.Overexpression of central CART through i.c.v. injections at either the forebrain (LV), interbrain (3V) or hindbrain (4V) areas has conformed to an anorexigenic role of CART, while the appetite-promoting effects of CART administered into specific neuronal targets may be attributed to a role of hypothalamic CART in stimulating the local release of orexigenic neuropeptides in a hypothalamic nucleus-specific manner. The discrepancy in anorexigenic and orexigenic circuitry highlights the major pitfall of non-specific widespread effects associated with i.c.v. ligand delivery, and was recognized through the efforts to identify potential sites for CART action following the analysis of Fos expression pattern representative of i.c.v. CART-induced neuronal activities. Candidate sites that may house the key mediator for the hypophagic effects of i.c.v. CART primarily reside in various feeding-related areas, including major hypothalamic nuclei, specific brainstem structures, and the Acb of the cerebral striatum. The appetite-inhibiting and -promoting effects produced by intra-accumbal CART and CART knockout murine models, to determine effects of CART depletion on feeding behavior and metabolism. General consistency has been demonstrated between results from the literature and in-house data from our novel CART KO model, supporting CART as a satiety factor. Collectively, the phenotypic effects of CART deficiency from birth, although inconsistent across studies, display promotion toward positive energy balance. Extended findings stemming from our novel KO model also denote that CART deficiency may confer an advantage in body weight conservation during starvation, potentially associated with an improved stress response attributable to an altered anxiety-related cascade involving the CART system.In parallel with overexpression approaches, independent research groups have adopted genetic ablation to generate several germline CART knockout animals, as well as conditional, developmental stage, or whole brain CART knockout models. Nevertheless, temporal or tissue-specific genetic ablation serves an invaluable means to minimize common pitfalls associated with a germline knockout strategy, such as any secondary effects or compensatory mechanisms during development.Both gene knockout and overexpression studies of CART have consolidated the stimulatory role of CART in promoting lipid oxidation and inhibiting fat accrual. As discussed above, despite the widely documented anorectic evidence of CART, possibility of an orexigenic role is indisputable. Under certain circumstances, such postulation may provide explanation for the inconsistent or lack of marked phenotypes in terms of energy balance observed in germline CART gene, whereas mutation profiling has associated alterations in CART with reduced metabolic rate and resting energy expenditure, hyperphagia, obesity and elevated incidence of type II diabetes. Intriguingly, whilst a role of CART in regulating energy expenditure remains equivocal in overexpression studies and considered discordant in gene deletion approach in animal models, mutation screening in humans has consolidated the involvement of CART in the modulation of energy expenditure, body weight and dyslipidaemia.In human, the expression of CART transcripts and peptides has been characterized in various hypothalamic areas involved in appetite control and lipid homeostasis. From the perspective of population genetics, genome-wide study of human obesity-susceptibility loci revealed linkage to a locus harboring the human To elucidate the underlying cellular mechanisms, functional effects of mutations and polymorphisms that were linked to the development of obesity were investigated. The sequence alterations of interests were described to potentially interfere with the post-translational processing of CART, leading to defective protein biosynthesis and deranged cellular distribution of the peptide, while modulation in the nuclear protein binding affinity was also reported to attenuate biological activity. Despite the plausible challenges imposed on the translatability of principal observations from animal models to the less well-documented human system, aberrations in CART have been demonstrated in human studies to promote positive energy balance, endorsing a primary anorectic role of CART.In sum, the CART system remains a dominant player in the regulation of feeding, body weight and energy metabolism, hence a promising candidate for the development of anti-obesity therapeutics. However, elucidating the underlying mechanisms of CART action, developing relevant pharmacological tools and understanding the nature of the endogenous CART receptor(s), remains crucial in unraveling the functional role of CART in energy homeostasis and obesity.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Cancer is a leading cause of death worldwide and represents one of the biggest biomedical research challenges of our time. Tumor progression is caused by somatic evolution of cell populations. Cancer cells expand because of the accumulation of selectively advantageous mutations, and expanding clones give rise to new cell subpopulations with increasingly higher somatic fitness . In the In general, cancer research involves a range of clinical, epidemiological, and molecular approaches, as well as mathematical and computational modeling. An early and very successful example of mathematical modeling was the work of Nordling  and of ANext-generation sequencing (NGS) technologies and their various applications have changed this situation fundamentally . Today, Analyzing these complex data and interpreting them in the context of ongoing somatic evolution, disease progression, and treatment response is a major challenge, and the prospects to improve cancer treatment depend critically on progress with these computational and statistical tasks. In the following, we briefly summarize the current state of the art in the field and highlight major challenges that lie ahead, including (i) reconstruction of evolutionary history based on different types of genomic alterations, (ii) functional interpretation of mutations, and (iii) predictive modeling of the evolutionary dynamics of cancer. We argue that an interdisciplinary approach, including statistical and computational data analysis as well as evolutionary modeling of cancer, will be essential for translating technological advances into clinical benefits.Cancer is a genetic disease that arises when normal cellular functions are disrupted by mutations arising in DNA. These changes occur at the level of single cells, which are then propagated into subpopulations as cells divide and pass mutations through cell lineages . DiffereThe resultant intratumor genetic diversity is a huge problem for correctly diagnosing and successfully treating tumors ,11. For Epistatic interactions among mutations are abundant. For example, many different cancer mutations can result in deregulation of the same signaling pathways, and distinct mutational patterns may result in the same phenotype ,14. ConvIn order to assess tumor diversity and to better understand the evolution of cancer, paired-end sequencing experiments can elucidate the genetic makeup of tumors. A single sample will provide a snapshot of the end result of these evolutionary processes across the cells that are sequenced at that point in time. We would like to use this information to infer the evolutionary history of the tumor, evaluate the rates of mutation and selection, and predict future responses of the tumor to environments potentially controlled by various drug protocols.The detected mutations can take the form of single-nucleotide variants (SNVs), in which a single nucleotide substitution occurs , or structural variants (SVs), in which chunks of DNA are erroneously copied, deleted, or misplaced, which in turn can lead to copy number variations (CNVs). Epigenetic changes affecting chromatin conformation, such as DNA methylation or histone modifications, can also arise. Paired-end sequencing offers a means to obtain relatively comprehensive descriptions of all of this somatic variation .Even so, a single snapshot of a genome can only provide so much information. Modern sequencing techniques now enable analysis of spatial and temporal effects. For example, samples can be taken from different locations in a patient, either within a tissue or including primary tumor and distant metastases ,19. Suchhttp://cancergenome.nih.gov/) and the International Cancer Genome Consortium (ICGC) (https://icgc.org/). Both have collated open-access data for hundreds to thousands of samples available to the cancer research community for further study. The generation of these great volumes of data of different types is inevitably leading to a range of computational and statistical challenges.There have been concerted international efforts over the last few years to produce comprehensive libraries of cancer genome data across a range of tissues, including The Cancer Genome Atlas (TCGA)  oncogenes that, when gaining increased activity through mutations, directly promote cancer  and (2) tumor suppressor genes that normally protect against cancer . Tumor initiation models have highlighted the different dynamics of oncogene activation versus tumor suppressor gene inactivation, the role of chromosomal instability, and the importance of the spatial structure of the tissue of origin .Tumor progression models focus on the process of mutation accumulation and further neoplastic transformation in an initiated tumor. These models have been used to infer the velocity at which mutant waves sweep through the cancer cell population and to elucidate how this speed of adaptation depends on the mutation rate, the fitness advantage of driver mutations, and on the feasible set of mutational pathways –60. The Mathematical models of drug resistance development date back to Luria and Delbrück , who stuMathematical modeling of tumor cell population dynamics will result in models and software tools that are potentially predictive of disease progression and treatment outcome. At present, much of the large-scale sequencing that has taken place is of limited depth and geographical scope, and mathematical models based on such information will likely be crude and approximate. To construct realistic models of the evolutionary processes taking place, we need to combine models that are representative of the processes taking place with high-resolution data. In the long run, ecological models of the entire tumor microenvironment may play this role . The besAPC, KRAS, and TP53 .–74.71–74Cancer progression models have been shown to improve prediction of patient survival. In particular, they allow for quantification of the degree of progression for each tumor, for example, as the expected waiting time for the specific mutational pattern to accumulate ,78. As sDriven by technological advances in genomics, cancer research is changing rapidly. Mathematical, statistical, and computational methods play an increasingly important role in this process, and many problems that occur in today’s cancer research can be addressed with methods that are familiar to computational biologists. On the other hand, many novel modeling and computational problems arise in cancer research that lead to methodological research in computational biology. With the ongoing large-scale generation of  genomic profiles of tumors that are publically available, computational biologists have the opportunity to make substantial contributions to cancer research. Here, we have highlighted a few major challenges which we believe are central for making progress, including (i) reconstruction of the evolutionary history of a tumor based on different types of genomic alterations, (ii) functional interpretation of mutations, and (iii) predictive modeling of the evolutionary dynamics of cancer. Statistically robust and computationally efficient methods of addressing these challenges will enable a range of applications, such as optimal control of tumor development, forecast of drug resistance evolution, and design of optimal individualized treatment strategies.


The maintenance of glucose homeostasis is complex and involves, besides the secretion and action of insulin and glucagon, a hormonal and neural mechanism, regulating the rate of gastric emptying. This mechanism depends on extrinsic and intrinsic factors. Glucagon-like peptide-1 secretion regulates the speed of gastric emptying, contributing to the control of postprandial glycemia. The pharmacodynamic characteristics of various agents of this class can explain the effects more relevant in fasting or postprandial glucose, and can thus guide the individualized treatment, according to the clinical and pathophysiological features of each patient. Besides primarily metabolic actions, this treatment affects other systems and, therefore, has aroused increasing interest. One of these actions is the reduction in gastric emptying rate.Incretin-based treatment  involves the secretion of insulin by a glucose-dependent mechanism. This action, along with the reduction in the levels of glucagon, have emphasized the need to customize the treatment approach for diabetes, basing treatment targets and medication on the pathophysiologic aspects peculiar to each case. Reviewing these foundations is, therefore, an essential task.Recently, the American Diabetes Association and the European Association for the Study of DiabetesThe gastric emptying rate is variable and depends on the content of the meal. Thus, if the fluid ingested is rich in glucose or calories, emptying is delayed. This inhibition determines a constant rate of energy absorption, of approximately 2 to 3kcal/minute; glucose infusion in the duodenum inhibits gastric emptying proportionally to the amount infused. This sensitive mechanism depends on extrinsic and intrinsic factors. depends on neural and hormonal feedback induced by the interaction of nutrients in the lumen of the small intestine, and is calorie-dependent. When food reaches the bowel, L and K cells of the distal small intestine produce GLP-1 and glucose-dependent insulinotropic peptide - GIP, which act on several tissues. In the hypothalamus, the peptides reduce appetite and send cholinergic and peptidergic signs to the vagus, inhibiting antral motility and stimulating pyloric motility. These actions contribute to the inhibition of gastric emptying. GLP1 and GIP stimulate insulin secretion and inhibit glucagon secretion. Moreover, they stimulate secretion of an amyloid pancreatic peptide, amylin. Insulin has a satiety effect on the central nervous system, while amylin decreases gastric emptying, due to vagal action.The extrinsic path, also known as “ileal brake mechanism”, Thus, in addition to the calorie aspect (extrinsic), blood glucose variations (intrinsic) may, through hypothalamic actions, increase or decrease appetite and activate the parasympathetic system that controls gastric emptying. This delicate neural and hormonal interaction prevents postprandial hyperglycemia in normal individuals.Intrinsic mechanisms involve effects of hyperglycemia or hypoglycemia. Hyperglycemia stimulates secretion of insulin and amylin, reducing the secretion of glucagon; it also decreases the secretion of ghrelin, which reduces gastric emptying, via parasympathetic signal. Physiologically, ghrelin increases the gastric emptying rate. and type 1 and 2 diabetic individuals. It was suggested initially that hyperinsulinemia also delays gastric emptying, but recent studies in type 1 diabetic individuals,19 have challenged these observations.Some studies showed that pronounced hyperglycemia (>250mg/dL) affects motility of the esophagus, stomach, small intestine, colon and gallbladder, both in normal Hence, higher overloads lead to more intense elevations in the levels of GLP1, resulting in progressive reduction in gastric emptying rate, a physiologic mechanism that contributes to blood glucose homeostasis. These data were reproduced in individuals with well-controlled type 2 diabetes, who received intraduodenal infusions of increasing amounts of glucose. This important study demonstrated that concentrations of insulin, GLP1 and GIP increased with intraduodenal overload of glucose, both in normal and diabetic individuals.In healthy individuals, the behavior of blood glucose and incretin levels is highly dependent on the exposure of the small intestine to carbohydrates.Preservation of this control mechanism in patients with diabetes opens possibilities of treatment with strategies that influence the rate of duodenal exposure to carbohydrates. made it clear that postprandial hyperglycemia is an independent risk factor for macrovascular disease, and its control is essential for reducing cardiovascular mortality in diabetic individuals. The approaches for controlling postprandial glucose levels include prandial insulin analogues, acarbose – of which action occurs, in part, due to increased levels of GLP1, pramlintide – of vagal action and, more recently, treatments based on the incretin system, especially GLP1.The DECODE study The concomitant use of prokinetic agents, such as metoclopramide, domperidone, cisapride and erythromycin, blocks the effect of GLP1 to control postprandial blood glucose, demonstrating the importance of its influence in gastric emptying.The mechanism of action of GLP1 during fasting essentially involves the secretion of glucose-dependent insulin and inhibition of glucagon. Recent studies have demonstrated, however, that its action in the postprandial state occurs through deceleration of gastric emptying, leading to reduced entry of glucose in circulation. This is due to induction of tachyphylaxis at the vagus level, and causes an attenuation of the response after chronic administration.Continuous exposure to high concentrations of GLP1 leads to a pronounced loss of ability to decrease gastric emptying., nor to the desensitization of the effect on insulin secretion and fasting blood glucose control. On the other hand, molecules, whose interaction with the receptor is short have greater effect on the reduction of gastric emptying – and, consequently on the postprandial glucose control – because they do not lead to desensitization.The pharmacokinetic and pharmacodynamic analyses of GLP1 demonstrate that this class of drugs is comprised of molecules with variable binding profiles to receptor of GLP1, which probably explains some differences of action among them. The prolonged action with the receptor leads to desensitization of the inhibitory gastric effect, but neither due to the anorectic effect that occurs through hypothalamic GLP1 receptorsversus more prolonged (liraglutide) action shows differences in their impact on the regulation of fasting and postprandial blood glucose levels, and may guide individualized treatment, according to the clinical and pathophysiological characteristics of each patient.The comparison between agents with faster (lixisenatide) We are heading toward treatment of diabetes based on individualization of medications. The choice of the best pharmaceutical will be based on understanding the pathophysiology of each case and the mechanism of action of medications. GLP1 analogues with prolonged action may benefit patients with high fasting glucose levels and who need to lose weight. Fast action analogues will be a very good option to correct cases of postprandial hyperglycemia.