In vivo research is critical to the functional dissection of multi-organ systems and whole organism physiology, and the laboratory mouse remains a quintessential animal model for studying mammalian, especially human, pathobiology. Enabled by technological innovations in genome sequencing, mutagenesis and genome editing, phenotype analyses, and bioinformatics, in vivo analysis of gene function and dysfunction in the mouse has delivered new understanding of the mechanisms of disease and accelerated medical advances. However, many significant hurdles have limited the elucidation of mechanisms underlying both rare and complex, multifactorial diseases, leaving significant gaps in our scientific knowledge. Future progress in developing a functionally annotated genome map depends upon studies in model organisms, not least the mouse. Further, recent advances in genetic manipulation and in vivo, in vitro, and in silico phenotyping technologies in the mouse make annotation of the vast majority of functional elements within the mammalian genome feasible. The implementation of a Deep Genome Project—to deliver the functional biological annotation of all human orthologous genomic elements in mice—is an essential and executable strategy to transform our understanding of genetic and genomic variation in human health and disease that will catalyze delivery of the promised benefits of genomic medicine to children and adults around the world.A comprehensive understanding of genetics, at the single locus, gene, and genomic level, and the pathophysiological consequences of gene variation resulting in gene, RNA, or protein dysfunction are crucial to meeting societal expectations of precision medicine and critical to optimizing clinical practice. With over 80% conserved synteny and a high degree of gene orthology, the mouse and human genomes have provided a unique opportunity for comparative functional analysis and the use of genetically altered mice to interrogate the pathobiology of human disease . For exawww.monarchinitiative.org) that use controlled vocabularies to integrate numeric, text, and image biological information from heterogeneous datasets  link genotype to phenotype and enable comparisons between mouse and human ontologies [CRISPR/Cas9 has enabled rapid and highly efficient targeted mutagenesis of the mouse genome. Concurrent development of in vivo analytical and imaging technologies has transformed high-throughput pipelines for precise and reproducible phenotyping of mouse mutants . Deep phtologies . These awww.mousephenotype.org). The magnitude of this global effort reflects the spirit and scale of the Human Genome Sequencing Project. The IMPC uses homologous recombination in embryonic stem (ES) cells and CRISPR/Cas9 technology to create mutants for genes in the mouse genome followed by whole organism phenotyping of female and male cohorts of adult mice and embryos [Realization of this goal is the foundation of the work of the International Mouse Phenotyping Consortium (IMPC). The IMPC is a coordinated program of 20 research laboratories in 12 countries on 5 continents dedicated to the design, production, and description of the function of human gene orthologs in the mouse genome ( embryos . The foc embryos  recognizhttps://www.mousephenotype.org/data/publications). This work continues on an industrial scale generating novel insights into gene-based disease phenotypes and other scientific domains such as conservation and ecology. This combined mouse production, phenotyping, and informatics approach has recently been applied to ~ 1/3 of known Mendelian disease genes and detected significant phenotypic similarities between human disease genes and mouse knockouts  of the orthologs for approximately half the genes [Gene association with a broad diversity of human diseases, including hearing loss, ocular diseases, metabolic disorders, bone pathologies, developmental abnormalities, and others, differentiated by sex, has been revealed through IMPC-led discovery research and IMPC-fueled studies by the broader scientific community (he genes . At leashe genes .https://www.mousephenotypetest.org/about-impc/).Currently, IMPC has generated null mutations for nearly 9000 genes of which over 6000 have been phenotyped. By July 2021, IMPC will complete comprehensive phenotypic annotation for over 9000 genes, representing about half of the ~ 18,000 human orthologs in the mouse , the Undiagnosed Diseases Network (https://undiagnosed.hms.harvard.edu/), Canada’s Care4Rare (http://care4rare.ca), The Gabriella Miller Kids First Pediatric Research Program (https://kidsfirstdrc.org/), the Genomics England Project (https://www.genomicsengland.co.uk/), and the “Fondation Maladies Rares” . Animal model data, including mice, facilitate the interpretation of potential causal variants among variants of unknown significance in clinical sequencing.If revealing the full biological role of every gene is not daunting enough, the pathobiological effects of individual human genetic and genomic variation further escalate the challenge. As exome sequencing (ES), clinical exome sequencing (cES), and whole genome sequencing become more commonly used in research and medical diagnostics to establish an etiologic molecular diagnosis, the number of variants of unknown clinical significance  is increasing exponentially and exceeding our current capabilities to interpret loss-of-function alleles . ImportaAlthough substantial efforts to date have revealed a fuller understanding of the functional landscape of the entire mammalian genome, significant and important knowledge gaps remain that limit the ability to interpret the causal relationship of genes and genetic variations to human development and disease. For instance, the majority of published gene to phenotype studies continue to focus on genes that are well-annotated or for which knowledge of biological function and pathological consequences of mutations already exist . As a reOnly around 21% of the ~ 20,000 genes annotated on the reference human genome have variation in them tied to a human disease trait  is also formulated around these four steps.Complete functional annotation of the protein-coding genome. Complete loss-of-function mutations are essential for identifying the phenotypic impact of protein-coding genes and a necessary first step to interpreting clinically relevant human genetic variation causing disease. By 2021, at the conclusion of the IMPC’s current mandate, ~ 9000 human orthologous genes in mice will remain to be analyzed by the consortium. Stopping at this point, halfway through the genome, would be equivalent to the Human Genome Project halting its sequencing effort after assembling euchromatic sequence of just 11 chromosomes. It will be vital to continue efforts toward the completion of the functional analysis of the remaining unannotated protein-coding genes using mouse models.Establish functional evaluation of the noncoding genome. The entire coding region is only 3–5% of the mammalian genome. The remaining 95% of the genome plays many roles across a variety of biological processes, including DNA replication, transcriptional regulation, and genomic structure. Of particular note, variation in enhancers, silencers, promoters, and insulators can have significant impact on both normal and abnormal gene expression [Strategies must be implemented for the prioritization and modeling of mutations of conserved noncoding elements in order to fully explore the in vivo function of the darkest part of the genome.pression  and genepression . StrategTranslate functional biological knowledge to clinical knowledge. The emerging field of genomic and precision medicine relies on the ability to interpret the potential pathophysiological consequences of genetic variation in patients. Beyond academic and research considerations, the long-term financial investments in translating human genetic variation to functional phenotypes and disease mechanisms is enormous and growing. Global investments from government agencies and corporate investors are predicted to nearly triple from US$79billion to over US$200billion in 10 years (https://bisresearch.com/industry-report/precision-medicine-market.html). For example, programs from the US All of Us Project, UK Biobank, the UK 100K Genome Project, the Chinese Precision Medicine Initiative, and many others are enrolling volunteers in efforts to gather personal history, clinical information, genome sequences, and environmental metadata to understand the role of genes in health and disease in order to identify new targets for molecular therapies. For these investments to deliver on their promise to improve health outcomes from molecular diagnosis to management and therapeutic intervention, it will be necessary to integrate gene function data generated from the study of mouse and other animal models into clinical databases, such as ClinVar, ClinGen, and others. Continued development of model organism databases, along with improvements in data integration and analysis, is needed to enable mechanistic insight into genetic variation and disease and support future developments in genomic and precision medicine.Enable rapid functional assessment of genomic variation and integrate functional testing into the clinical decision-making process. While statistical inference of human patient data is currently used to discriminate disease causing from benign associated variants, a definitive molecular diagnosis is often not attainable. Even in those cases where this approach is sufficient, the delay in diagnosis is costly—psychologically, socially, and economically. It will be necessary to undertake programs for the rapid creation and analysis of mouse models of human coding variants along with more efficient approaches to phenotyping. These models, along with other mutational variants, will inform diagnostic decisions and targeted treatments. With these programs in place, clinicians and their research colleagues could rely on mouse models as diagnostic and therapeutic testing platforms, examine the pathological significance of a genetic variant in an orthologous mammalian system, interrogate gene/phenotype relationships for different types of alleles , explore potential gene/environment effects, and access comprehensive datasets to help guide clinical decision-making. In turn, mouse genetic experts will need to respond quickly with targeted phenotyping of mouse models in order for them to achieve clinical utility. It will be necessary to optimize funding and bring scientific insights from mouse functional data to inform the application of mouse models as human patient avatars, make the knowledge gained from mouse data available via the electronic medical record, and enhance education and training of clinicians in genetics and genomic medicine.We postulate that four steps undertaken by the collective endeavors of the global community will be needed to drive progress in genomic and precision medicine. These four steps, allied to genome-wide goals for in vitro systems and other model organisms, will deliver a deeper and more comprehensive understanding of individual gene function, make biological resources and data available to experimentally decipher disease mechanisms, interpret genomic variation, and reduce the diagnostic odysseys of patients with variants of unknown significance. The IMPC’s strategy 2021–2030 (Despite the incredible scientific advances in genetics at the single gene and genomic level, the collective biomedical community has only begun to scratch the surface of knowledge about the diverse and varied in vivo pathobiological roles of functional elements throughout the human genome. The global mouse genetics community is primed to address the grand challenges that we face to fully comprehend the role of genes and genetics in development, biological homeostasis, systems biology, disease, and medicine, and is ready to launch a new era for the systematic study of the function of the mammalian genome. The time is right to embark on a Deep Genome Project, on the scale of the Human Genome Project, that will fundamentally enhance the knowledgebase across the biomedical sciences. Driven by the enormous potential of mouse genetics and allied to developments in other model organisms and in vitro approaches, this project will be transformative for biology, medicine, and global health.


We assessed the risk of fatal occupational injuries among migrant workers with two different types of employment permits in South Korea. This observational study used national data from January 2007 to September 2018 and analyzed 42,089 cases of occupationally injured migrant workers of Chinese nationality. Fatality rates were analyzed according to year, sex, age, occupation, industry, and type of employment permit. Chinese workers were permitted to work for one employer and prohibited from changing employers, whereas Korean-Chinese workers were permitted to change their employer. The adjusted fatality rate of occupational injuries of Chinese migrant workers was significantly higher  than that of Korean-Chinese migrant workers. The prohibition on changing workplaces; male sex; age ≥ 45 years; machine operator; construction industry; and agriculture, livestock, and fisheries industry were risk factors for fatal occupational injuries. The results imply a need for revision of the migrant-worker employment permit systems and implementation of occupational safety and health policies for all workers to promote health equity. Worldwide, 2.3 million workers die annually due to occupational injuries and diseases . MigrantAlthough the occupational safety and health (OSH) of migrant workers is a global issue, only a few studies have investigated the related risk factors. While not specialized for migrant workers’ OSH, a “conceptual model for integrated approaches to protection and promotion of worker health”  helps unSouth Korea is one of the major destination countries for migrant workers in Asia. Two kinds of permits are available under the Employment Permit System (EPS), a dependent employment permit and a work permit, although the latter are not common in South Korea. Migrant workers in other major destination countries usually work with a work permit. The policy of prohibiting a change of workplace is a distinguishing factor of working conditions for migrant workers who hold a dependent employment permit. Such migrant workers may not work for another employer unless the current employer breaks the law or the business closes . The pron = 215,665) of documented migrant workers were of Chinese nationality [The two migrant-worker employment permit systems can be evaluated by comparing groups of migrant workers of Chinese nationality with similar migration histories. The work permit system only applies to workers of Korean descent and Korean Chinese people account for 95.7% of all workers of Korean descent . In Septionality . Korean-ionality . In the Korean-Chinese and Chinese migrant workers may also differ in terms of their ability to change jobs, Korean language proficiency, and whether they have relatives in South Korea. These factors may impact the fatality rate of occupational injury (FROI). Also, previous studies cite a lack of labor rights, restricted access to family and other support systems , and theIn this study, we determined whether the FROI differed due to prohibition on changing workplaces between Korean-Chinese and Chinese migrant workers. In addition, we identified the risk factors for fatal occupational injuries among Chinese migrant workers in South Korea. This is the first study of the effects of the prohibition on changing workplaces on the FROI of migrant workers.We used data from the Korea Workers Compensation and Welfare Services (KCOMWEL). The KCOMWEL is a national agency that manages all claims for compensation due to occupational injuries and diseases. Raw de-identified data were obtained from the KCOMWEL website through an information request system . The datThe annual data comprised work-related injuries and diseases with fatal and non-fatal outcomes, causes and sites of injuries, demographic variables , and employment variables . According to the KCOMWEL definitions, occupational injuries were defined as those that occurred while workers were engaged in work-related activities, including commuting (after 2018), that required >4 days of medical treatment. From 2007 to 2018, there were a total of 71,593 occupational injury or disease compensation claims, of which 42,089 were filed by migrant workers of Chinese nationality. The exclusion criteria were as conservative as possible to ensure generation of reliable results. Selection of the study population is shown in The study population comprised migrant workers of Chinese nationality whose workers’ compensation claims due to occupational injuries were approved. We defined migrant workers as persons of non-Korean nationality working under the EPS irrespective of their legal status. The Ministry of Employment and Labor of South Korea has, as of September 2018, issued memoranda of understanding on migrant workers with the governments of China, Vietnam, Sri Lanka, Uzbekistan, Indonesia, Thailand, the Philippines, Nepal, Mongolia, Pakistan, Bangladesh, Cambodia, Myanmar, Kyrgyzstan, East Timor, and Laos . As of tThis observational study evaluated risk factors for fatal occupational injuries by comparing the FROIs of two groups of migrant workers of Chinese nationality. The FROI is defined as the proportion of occupational deaths among migrant workers as a result of occupational injuries. We used FROI as the outcome variable because the death count was the most reliable measure due to the high rates of under-reporting, particularly by migrant workers in South Korea. According to the OSH Research Institute of South Korea, over 70% of migrant workers who suffered an occupational injury failed to file claims for workers’ compensation . A poor Korean-Chinese migrant workers have the right to employment of their choice under the working-visit system of the EPS. To assess the effect of the prohibition on changing workplaces, we compared the FROIs of Korean-Chinese and Chinese migrant workers with adjustment for year, sex, age, occupation, industry, and site of injury. Since the start of the EPS, there have been two revisions to the law regarding the prohibition on migrant workers changing workplaces, one on 9 October 2009 and the other on 1 February 2012. Migrant workers with a dependent employment permit may apply for a workplace change only for special reasons. The 2009 revision slightly expanded the “special reasons,” and the 2012 revision clarified these reasons by providing examples; the restriction on changing workplaces remained in place. We divided the study period into three parts by the years in which revisions were issued to also assess the longitudinal effect of the related policies.The employment of migrants is tightly controlled by the EPS with regard to both the annual numbers and the types of positions. The occupations classified by the seventh Korea Classification of Occupational Standards were divided into the four employment categories in which migrant workers were most frequently employed: manager and service workers; agriculture, livestock, and fisheries workers; machine operators; and elementary workers. Managers and service workers were grouped together due to the low fatality rates  in theseNine industries were classified into five categories in which migrant workers could be employed: manufacturing, construction, service, agriculture, livestock, and fisheries. However, because there were few claims for workers’ compensation from the agriculture, livestock, and fisheries industries, we treated these as a single unit-agriculture, livestock, and fisheries (ALF). Furthermore, the transportation and delivery industries were classified as other, and the sales, food, and accommodation sectors were pooled as the service industry, in which Korean-Chinese workers account for 93.3% of the migrant workers . The sitThe data from 42,089 migrant workers of Chinese nationality were analyzed in three stages. First, a chi-squared test was used to determine whether the FROI differed significantly according to the characteristics of the study population. Second, differences in the FROIs of each covariate were compared between the Korean-Chinese and Chinese migrant workers. Third, we identified risk factors for fatal occupational injuries using three logistic regression models with adjustment for covariates. R software (version 3.5.1)  was used to conduct all statistical analyses.The FROIs of the study population are listed in There were differences in the causes of death due to occupational injuries according to the group and industry variables . CollisiThe FROIs of Chinese migrant workers according to their characteristics were equal to or greater than those of the Korean-Chinese migrant workers . The aveThe differences in FROI between the two groups according to year allowed us to gauge the effect of the two revisions to the law regarding the prohibition on changing workplaces. Unlike Korean-Chinese migrant workers, the FROI of Chinese migrant workers did not show a decreasing tendency over time. The FROI of Chinese migrant workers decreased after 2009, when the first revision occurred, but increased after 2012, when the second revision took place. Based on the different trends in the FROI between the two groups, we evaluated the interaction effects between group and year, controlling for other variables  . The intThe FROI of Chinese migrant workers was significantly higher  than that of Korean-Chinese migrant workers  after coThe FROI did not differ according to age until subjects were in their mid-40s. Compared to subjects 15–28 years of age, the FROIs of those aged 45–53 years and 54–81 years were more than two-fold higher. Thus, age ≥ 45 years was a risk factor for fatal occupational injury. Machine operators had a significantly higher  FROI than managers and service workers. The ORs of the construction and ALF industries were significantly higher using the service sector as the reference industry. Therefore, machine operating and the construction and ALF industries were associated with fatal occupational injuries among migrant workers.We evaluated factors associated with fatal occupational injuries among migrant workers in South Korea using data from 42,089 Chinese migrant workers from January 2007 to September 2018. The results showed that the prohibition on changing workplaces; male sex; age ≥45 years; machine operator; and the construction and agriculture, livestock, and fishery industries were risk factors for occupational injuries among migrant workers.A high FROI may indicate the occurrence of occupational injuries of greater severity. Whole-body injuries were overwhelmingly fatal . ChineseThe FROI of Chinese migrant workers after controlling for the potential confounding effects of covariates was 1.80-fold higher in Model 3 and 1.22-fold higher in Model 1 than that of Korean-Chinese workers. The difference in FROIs between the two groups was tied to our second research question, namely, identifying the risk factors of occupational fatal injury. The following parameters need to be considered when comparing the two groups: (1) Korean language proficiency, (2) the presence of family and relatives in South Korea, and (3) the prohibition on changing workplaces.First, the language barrier is not an important issue in South Korea because all migrant workers, including Korean-Chinese migrant workers from EPS countries, are required to score well on the official Employment Permit System Test of Proficiency in Korean before beginning employment. Although Korean-Chinese workers have better proficiency in the Korean language than do Chinese migrant workers, Chinese migrant workers achieve higher scores than EPS workers from other regions , which pSecond, the right to family reunification for migrant workers in South Korea is restricted. Until 2007, Korean-Chinese migrants could obtain a labor visa only if they were invited by relatives residing in South Korea; since then, they can be granted a visa without an invitation. However, such visas do not allow migrant workers to bring their families with them. It is important to note that access to family support is not only a basic human right but also an important protective factor from occupational injury . The KorTherefore, the prohibition on changing workplaces, rather than language proficiency and access to family, likely explains the disparity in the incidence of fatal occupational injuries between Korean-Chinese and Chinese migrant workers. Additionally, the interaction effect between group and year  supportsThe prohibition on changing workplaces has a negative impact on the health of migrant workers, and its maintenance lacks a scientific basis. According to the precedent statement and the International Organization for Migration (IOM) report, the provision exists for economic reasons. The EPS was instituted to address manpower shortages at small and medium-sized enterprises, and the South Korean government accepted employers’ claims that they would suffer from wage hikes . HoweverExpanding the scope of the law regarding changing workplaces could affect the FROI of migrant workers . PermissInjured male migrant workers were more likely to die than were injured female migrant workers. However, in South Korea, only workers who meet the criteria of the Labor Standards Law can claim workers’ compensation under the Korea Workers’ Compensation Insurance Act. It is thus possible that some female migrant workers were unable to claim workers’ compensation because they were not legally defined as workers. In the case of domestic workers who perform domestic duties such as cleaning, cooking, and looking after children or elderly people , the oveAge ≥45 years was a risk factor for fatal occupational injuries among migrant workers, as reported previously . In contn = 26) fatalities, suggesting that ALF workers could not claim workers’ compensation unless they were severely injured.Machine operation was the highest-risk occupation, and construction and ALF were the most physically demanding industries. Only machine operators had a significantly high risk of occupational injury, compared to managers and service workers. Construction and ALF were risk factors for fatal occupational injuries, consistent with previous reports ,37. The This study has several limitations. First, persons not defined as workers according to South Korean law are omitted from the workers’ compensation claims data; thus, further studies that include such workers are needed. Second, the occupational categories may have been inaccurate. The colleagues or agents who reported deaths may have misclassified the occupation of the deceased . DespiteA longitudinal follow-up study of the health of migrant workers is needed to ascertain whether the prohibition on changing workplaces affects not only occupational injuries but also occupational diseases. Because this was basically an observational cross-sectional study, we could not control for missing or unmeasured factors, and were unable to establish causality. The importance of this study lies in its comparison of recruitment systems and it being the first investigation of the effect of the prohibition on migrant workers changing workplaces on fatal occupational injuries.We compared two groups of migrant workers of Chinese nationality with different types of employment permits under the EPS. Of the two types of permits, it was found that the FROI for the group with the dependent employment permit, which prohibits changing workplaces was significantly higher. The status of occupational injuries and diseases of migrant workers’ data revealed the prohibition on changing workplaces to be a risk factor for fatal occupational injury. Additionally, male sex; age ≥45 years; machine operator; construction; and the agriculture, livestock, and fishery industries were associated with fatal occupational injuries among migrant workers in South Korea. Policies to improve the recruiting system and to strengthen workplace safety measures for better OSH of migrant workers are the responsibility of the global community as well as local governments and businesses. Ultimately, preventing occupational injuries to migrant workers in high-risk occupations will improve the health of all workers.


Antibiotics are cytotoxic or cytostatic compounds, very effective, harmful and specific against pathogenic microorganisms that have saved millions of lives and increased human life expectancy and quality  pathogens  and biofilm-forming organisms, a significant lower outcome of resistance development, reduction of single doses and a decrease in side effects  with carbapenems. Such antibiotics belong to the class of broad-spectrum antimicrobials routinely used for the treatment of infections caused by multidrug-resistant P. aeuruginosa that leads to chronic diseases. Recently, a novel synthetic cyclolipopeptide analog of polymyxin (AMP38) was tested in combination with carbapenems, and the synergistic effect was observed to cause the killing of biofilm-forming and carbapenem-resistant P. aeruginosa , P. aeruginosa, and Escherichia coli (E. coli) , P. aeruginosa and Acinetobacter baumannii (A. baumannii), especially in combination with rifampin  pathogens, revealed the ability of AMPs to elicit degradation of ppGpp, avoiding the entry in an energy-starved state. This is a significant finding as potentially all microorganisms react to antibiotic treatment mediating persistence, and the fact that some AMPs can prevent it opens new doors for the development of alternative therapies that effectively decrease the resistance rate  from infected lungs, liver, and spleen, more efficiently in combination with the human neutrophil peptide (HNP)-1 in comparison to when they were employed alone , a database for therapeutic peptides and proteins, only 4 % of all approved therapeutic peptides and proteins are administered orally. The following sections focus on the stability issue and the low membrane permeability of peptides in general and present some successful strategies that overcome the practical limitations of peptides as orally administered drugs.In clinical therapy, the most desirable route of drug administration is orally due to the relatively low cost of production and patient compliance , and as clinical evidence has shown, its low oral bioavailability allows the gastrointestinal tract concentrations to considerably exceed its MIC for the pathogen (Knight-Connoni et al., In addition to the stability problem, oral delivery of AMPs is also challenging due to their poor cell membrane permeability. Once orally administered, AMPs should cross the gastrointestinal epithelium to reach the bloodstream. However, this is not so simple for hydrophilic molecules exceeding 700 Da (Fosgerau and Hoffmann, The combinatory therapy AMP-Antibiotic has increasingly attracted attention within contemporary studies due to its diverse benefits. The number of combination studies involving AMP-antibiotic has therefore been exponentially growing over the last few years (Jorge et al., LP-C and NO-S are doctoral students who contributed to the preparation of the article. All authors contributed equally to this work.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.


Stenosis or obstruction of the celiac artery (CA) is known as celiac artery stenosis (CAS) and is usually accompanied by the formation of arterial anastomosis between the superior mesenteric artery (SMA) system and the CA system. Arterial bypass is mainly achieved through the gastroduodenal artery (GDA); therefore, the division of the GDA during pancreaticoduodenectomy (PD) could pose a problem in patients with CAS.We reported a case of PD presenting complete occlusion of the CA, in which perfusion to organs in the CA system was maintained via peri-pancreatic arterial communication. There were complicated arterial anastomoses around the pancreas, which were clearly visualized on a three-dimensional reconstruction of the arterial system using multi-detector computed tomography. Among these complicated anastomoses, one well-developed anastomosis between the SMA and the splenic artery through the dorsal pancreatic artery (DPA) was identified. The DPA was considered to work as a potential collateral pathway from the SMA to organs in the CA system after division of the GDA. During surgery, Doppler ultrasonography detected hepatopetal arterial flow even after the GDA clamping; therefore, we performed typical PD with division of the GDA. The postoperative course of the patient was uneventful, and there was no sign of ischemic complications in the CA system organs including the liver, stomach or spleen.Three-dimensional reconstruction of the arterial system using multi-detector computed tomography and the intraoperative GDA clamping test were useful to determine whether it was possible to divide the GDA in PD, in the case of CAS. The celiac artery (CA) is the first major branch of the abdominal aorta supplying blood to the liver, stomach, spleen, duodenum, and pancreas. Stenosis or obstruction of the CA, known as celiac artery stenosis (CAS), can result from various factors, including median arcuate ligament syndrome (MALS), atherosclerosis, and invasion of malignancies or pancreatitis . HoweverPancreaticoduodenectomy (PD) is a surgical procedure to resect pancreatic head tumor, including pancreatic adenocarcinoma, distal bile duct cancer, and ampullary carcinoma . The proAn 80-year-old man with epigastric pain visited our hospital in January 2019. A magnetic resonance cholangiopancreatography revealed several stones in the common bile duct (CBD), and he was diagnosed with choledocholithiasis. Endoscopic lithotripsy was performed, and cholangiography showed narrowing and wall-irregularity of the lower CBD Fig. a. A 3D iPD was performed in April 2019. After taping of the GDA Fig. , CHA, anPostoperative laboratory data showed slight elevation of hepatic enzyme immediately after the surgery Fig. , but we CAS is a relatively common finding, identified in 12.5–24% of patients undergoing abdominal angiography . The typSevere stenosis or obstruction of the CA often induces the formation of collateral arterial pathways from the SMA system to the CA system to compensate the decrease of splanchnic arterial flow of the upper abdomen. The main collateral pathway is reported to be the GDA or DPA in most cases, whereas various arteries, such as the DPA, jejunal artery, omental artery, left gastric artery, arc of Buhlar, and Riolan or Drummond artery can be a collateral pathway , 5. SongIn the context of CAS, the formation of collateral arterial pathways is surgically relevant for arterial division in PD. Although the GDA is the main collateral pathway from the SMA system to the CA system in most of the cases of CAS, the GDA must be divided in PD. Therefore, various procedures were attempted to avoid organ ischemia after PD in patients with CAS . There aIn conclusion, we described a case of PD with complete occlusion of the CA, in which we maintained perfusion via the peri-pancreatic arterial communication. A 3D reconstruction of the arterial system from MDCT and intraoperative GDA clamping test were useful to determine whether it was possible to divide the GDA.


Hepatocellular carcinoma (HCC) incidence is rising. This paper summarises the current state of knowledge and recent discoveries in the cellular and physiological mechanisms leading to the development of liver cancer, especially HCC, and liver metastases. After reviewing normal hepatic cytoarchitecture and immunological characteristics, the paper addresses the pathophysiological factors that cause liver damage and predispose to neoplasia. Particular attention is given to chronic liver diseases, metabolic syndrome and the impact of altered gut microbiota, disrupted circadian rhythm and psychological stress. Improved knowledge of the multifactorial aetiology of HCC has important implications for the prevention and treatment of this cancer and of liver metastases in general. Hepatocellular carcinoma (HCC) is an aggressive tumour of the liver. Among all cancers, HCC is one of the fastest growing causes of death globally . HCC is When HCC develops in liver parenchyma, it is encircled by extracellular matrix, stromal cells, and secreted proteins within an immunosuppressive tumour microenvironment (TME). The TME is populated by immune cells that interact among themselves and with the tumour. The makeup of the TME depends on the biological characteristics of the neoplastic cells and on the patient’s pathophysiological conditions. The immune system’s role in promoting HCC development has long been a focus of study ,8,9. ParHCC recurs after primary liver resection with curative intent in about 70% of patients . The risThis paper reviews the normal hepatic cytoarchitecture and immunological features, and then discusses factors that, by causing liver damage, predispose to the development of HCC and liver metastases. These factors include chronic liver disease, an altered gut microbiota composition, and changes in the circadian rhythm. The main physiological functions of the liver are metabolism of nutrients absorbed by the gastrointestinal tract and detoxification of dangerous molecules present in the circulation. These functions are responsible for a delicate balance between immune tolerance and immune response, and their correct functioning requires a normal cytoarchitecture of hepatic lobules . This cyThe vascular endothelial growth factor (VEGF) signalling pathway regulates vascular morphogenesis, which is important for both hepatic cytoarchitecture and cancer growth. The concentration of VEGF determines the directionality of the angiogenic sprouting, and vessel extension and growth (reviewed in ). VEGF eHSCs are also the main cell type involved in liver fibrosis, the formation of thickened, scarred connective tissue subsequent to liver damage. Activated HSCs sustained by a profibrogenic environment differentiate into collagen-producing myofibroblasts. These cells become detectable in the liver during liver injury. They produce an abundant extracellular matrix that initially accumulates in the space of Disse .Another important cell is the Kupffer cell, a self-renewing macrophage that, like the myofibroblast, accumulates in injured liver tissue. Kupffer cells recognise and clear pathogenic microbes and dying or dead host cells that arrive via the portal circulation. Depending on the local tissue environment, they adapt an M1 (pro-inflammatory) or M2 (anti-inflammatory) polarisation. During liver injury, Kupffer cells are activated to release mediators such as TGF-β and platelet-derived growth factor (PDGF), which regulate the activation of HSCs into myofibroblasts, promoting hepatic fibrosis . The liver, due to its anatomical location and structure, participates in the induction of tolerance to food. However, after activation of an immune response and inflammatory processes, for example to a neoplasm, hepatic macrophages and HSCs release large amounts of cytokines and adhesion molecules. These proteins contribute to the activation of endothelial cells, which facilitates the extravasation of neoplastic cells into the liver. These cells, in context of the tolerant liver environment, can develop into metastases  7,9,20,,20,21. Izonula occludens that separates the bile duct from the blood, thus forming a blood–bile barrier that restricts the mixing of blood and bile. Thus, preservation of normal membrane polarity is vital for hepatocyte functionality, such as apical bile secretion and simultaneous basal secretion of large quantities of serum proteins into the blood. The main factor involved in the formation and maintenance of hepatocyte polarisation is the Wnt/β-catenin signalling pathway  cells, contribute to improved survival after liver transplantation, and the finding that a decrease in immune defences, due to old age, immunosuppressive therapy or autoimmune disease, increases the risk of liver metastasis ,29,30. EThe liver contains numerous adaptive and innate immune cells that recognise foreign substances, recruit leukocytes, and are present to lymphocytes the antigens taken up from the circulation. In order to be effective, the immune system needs to be activated only against foreign molecules and not activated against host molecules; to do this, the immune response needs to be continuously controlled. The control between activating and inhibiting signals from the immune response is what characterises the liver as an immunologic organ . The livThe liver harbours several immune cell populations, each with its own specific function and hepatic location  34]. Ku. Ku34]. Activated Kupffer cells have been implicated in hepatocarcinogenesis . During Dendritic cells (DCs) are specialised antigen-presenting cells that play key roles in the initiation and direction of T cell responses. Antigens circulating in blood and lymph are captured by DCs, and then loaded DCs migrate through the lymph towards the lymph nodes, where they present the antigen to T cells. In the liver, DCs are found in the subcapsular space and around vessels such as the central veins . Their lGranulocytes, neutrophils and eosinophils migrate from the blood to the injury site in the liver, following gradients of chemokines from activated immune cells and of fragments of dead cells  . These cLymphocytes are enriched and naturally activated in the liver . Here, mThere are several hypotheses regarding the pro-metastatic state of a damaged liver consequent to cirrhosis, steatosis and nonalchoholic fatty liver disease. Some authors sustain that the altered hepatic cytoarchitecture, consequent to cirrhosis and steatosis, creates an unfavourable environment for the development of metastasis ,55. OtheSteatosis is an accumulation of triglycerides in hepatocytes. Risk factors for steatosis (and steatofibrosis) include metabolic syndrome and hepatitis virus infections. Steatosis is a reversible phenomenon, but when it persists, it modifies the hepatic cytoarchitecture. These changes in the architecture of the liver reduces the flow of blood and blood cells in the liver. Lipid accumulation, combining with ballooning degeneration, chronic portal inflammation, fibrosis, neoplastic microthrombi and an imbalance between the production and reduction of free radical fatty acids all contribute to creating a pro-inflammatory state with altered blood flow. In this setting, a nonalcoholic fatty liver disease (NAFLD) like steatosis develops into nonalcoholic steatohepatitis (NASH), causing fibrosis which results in cirrhosis. Traditionally, HCC was thought to develop mostly from cirrhosis. However, it is now clear that HCC as well as liver metastasis can also develop directly from inflammation without cirrhosis; this disease evolution is associated with metabolic disorders ,63. SeveNASH is a type of NAFLD characterised by steatosis, inflammation and necrosis. If NASH persists over time, the body’s attempts to repair the damage can lead to fibrosis by the replacement of hepatocytes with connective tissue. NASH can only be diagnosed by liver biopsy, but it can be suspected in patients with obesity, insulin resistance or high serum levels of transaminases; it is associated with a large production of ROS and the risk of HCC ,68. Oxidative stress, an essential factor in the pathogenesis of gastrointestinal mucosal diseases, was found to occur early in chronic liver diseases and in liver inflammatory responses. Consequently, the extetd of oxidative stress may reflect the degree of liver damage and hence the pro-metastatic predisposition of the parenchyma . MarkersThe intestinal microbiota consists of a variety of bacteria, viruses and fungi, present in different amounts, depending on the site within the intestinal tract and the subject’s age, ethnicity and state of health . The micThe interest in gut microbiota has grown recently with the discovery that it interacts with several extra-intestinal organs, including those of the cardiovascular system, the kidneys, brain, bone and especially liver . The comThe liver receives nutrients, toxins and microorganisms via the portal vein from the gut. When the intestine is inflamed, it is more permeable, so a greater amount of toxins and bacteria can reach the liver, favouring the development of inflammation in the liver ,78,79,80The circadian rhythm modulates most cellular functions in relation to the light–dark and temperature cycles caused by rotation of the Earth. The circadian regulation of metabolism also affects carcinogenesis, including that of the liver, as shown in a study in mice . A chronAccording to the Warburg theory of cancer, cancer cells tend to use anaerobic glycolysis (glucose fermentation) for metabolism instead of the more effective oxidative phosphorylation, because of insufficient cellular respiration in damaged mitochondria. Oxidative phosphorylation produces up to 36 ATP molecules per molecule of glucose, while anaerobic glycolysis results in just two ATP molecules. Consequently, neoplastic cells need an increased glucose uptake from the microenvironment and secrete more lactic acid, the end product of glycolysis. Lactic acid upregulates the expression of hypoxia-inducible factors (HIFs) and VEGF, which stimulate neo-angiogenesis . The lactate acidification of the microenvironment also has an immunosuppressive action favouring tumour immune escape . ExtraceCircadian rhythm also influences the complex neuronal network and the release of neurohumoural factors  . It has In recent years, the incidence and development of HCC has changed substantially due to the introduction of an anti-hepatitis B virus (HBV) vaccine and anti-HCV therapy. However, there has been no decline in the incidence of HCC associated with other diseases . A difference concerning HCC occurring in a background of or without cirrhosis also is emerging as a reflection of a difference in HCC pathogenesis. The common denominator of HCC risk among these different diseases, is the development and maintenance of a hepatic inflammatory state, in response to a viral infection, the accumulation of fat, or irritating molecules that reach the liver via blood and bile circulations. The fact that this inflammatory or pro-inflammatory state develops by different mechanisms extends the strategies that can be used at least theoretically, to reduce the risk of recurrence or metastasis. Of particular interest is the alteration of the cytoarchitecture and the microcirculation of the liver associated with the development of fibrosis, an increase in the inflammation of the hepatic microenvironment resulting, in particular, from a greater intestinal permeability to bacteria and toxins and the lipotoxicity associated with the metabolic syndrome, the immunosuppressive status occurring in the hepatic environment deriving from social stress, secretion of adipokines, alteration of the bile acid and the liver metabolism, which favour the engraftment of HCC. Of noteworthy the same molecular, metabolic and cellular processes, appears to favour also the development of recurrence and metastasis in the liver. The definition of specific pathways leading to liver damage offers the possibility of exploring therapeutic or prevention strategies to modulate the risk of disease recovery. Many of the conditions presented in this review could be modified by personalised therapeutic plans aimed at a personalised study of nutritional programs, the glycemic and lipid control, the modification of the intestinal flora and the increase of host immunocompetence, in addition to the obvious and already obsolete treatment of hepatic viral infections. Moreover, targeted therapies on distinct metabolic pathways involved in the progression of liver damage are now in development as reviewed in . Promisi


Diabetes mellitus (DM), a group of diseases characterized by defective glucose metabolism, is the most widespread metabolic disorder affecting over 400 million adults worldwide. This pathological condition has been implicated in the pathogenesis of a number of central encephalopathies and peripheral neuropathies. In further support of this notion, recent epidemiological evidence suggests a link between DM and Parkinson’s disease (PD), with hyperglycemia emerging as one of the culprits in neurodegeneration involving the nigrostriatal pathway, the neuroanatomical substrate of the motor symptoms affecting parkinsonian patients. Indeed, dopaminergic neurons located in the mesencephalic substantia nigra appear to be particularly vulnerable to oxidative stress and degeneration, likely because of their intrinsic susceptibility to mitochondrial dysfunction, which may represent a direct consequence of hyperglycemia and hyperglycemia-induced oxidative stress. Other pathological pathways induced by increased intracellular glucose levels, including the polyol and the hexosamine pathway as well as the formation of advanced glycation end-products, may all play a pivotal role in mediating the detrimental effects of hyperglycemia on nigral dopaminergic neurons. In this review article, we will examine the epidemiological as well as the molecular and cellular clues supporting the potential susceptibility of nigrostriatal dopaminergic neurons to hyperglycemia. In this age of effervescent neuroscientific developments, clinical profiles are more accurately drawn, long-term patient management is improving, and the pursuit of etiological and pathophysiological explanations is starting to bear fruit. The preferential degeneration of the dopaminergic nigrostriatal pathway is well appreciated today, and additional pieces of the puzzle are continuously falling into place. More importantly, the apparent link between diabetes (DM) and PD is rising momentum, with hyperglycemia representing the metabolic culprit linking DM with PD. Even though a handful of medical conditions may cause hyperglycemia, we deliberately hold our focus on DM as it offers a tangible collection of experimental and clinical evidence interlaced with PD pathophysiology.Since the publication of the pioneering report of Dr. James Parkinson, “An Essay on the Shaking Palsy,” where he described six subjects suffering from a pathological state that he termed The present review will lay out the epidemiological evidence linking DM and PD as well as the molecular mechanisms underlying the exacerbated susceptibility of nigrostriatal dopaminergic neurons to hyperglycemia, by illustrating glucose transport and metabolism in the brain and the role of glucotoxicity in promoting neuronal damage and degeneration. The transport kinetics and metabolism of glucose are highly complex, especially in the central nervous system (CNS), and bear significant implications in the initiation of oxidative stress, induction of non-enzymatic glycation and activation of biochemical pathways mediating the effect of glucotoxicity in neurons.PD is the prototype of movement disorders. In fact, the most prominent feature of PD is the presence of motor symptoms expressed in a majority of patients as a classical triad of resting tremor, bradykinesia, and rigidity  as a key contributor to movement performance  as a mitochondria-targeting agent of parkinsonism, broke new ground for the exploration of other chemicals, such as pesticides  upon auto-oxidation  than most other catecholaminergic neurons, implying the protracted presence of dopamine in the cytosol leading to the inevitable formation of toxic oxidative by-products and, ultimately, potentially toxic neuromelanin  ubiquitously present at the surface of cells accomplish the function of taking up glucose from the extracellular environment. In peripheral tissues, such as the skeletal muscle and adipose tissue, glucose uptake is mediated by GLUT4, an insulin-dependent glucose transporter. On the contrary, in the vast majority of the CNS glucose transport is mediated by insulin-independent GLUTs  which allows this transporter to work efficiently even at glucose concentrations surrounding the neurons in the order of 1–2 mM  axis  with more fuel, culminating in increased production of electron donors which are funneled toward the electron transport chain to fuel oxidative phosphorylation. In a second scenario, taking into account the possibility of an astrocyte-neuron lactate shuttle in hyperglycemia, lactate derived from glycolysis in astrocytes may enter the neurons and be converted to pyruvate to power the TCA. The end result is electron transport chain overload and subsequent increase in the proton gradient across the mitochondrial inner membrane which, in turn, impairs the transport of electrons across the mitochondrial complexes favoring the leakage of electrons and the formation of ROS .+-dependent DNA repair enzyme, poly (adenosine diphosphate-ribose) polymerase (PARP1), which, in turn, by depleting intracellular NAD+, in concert with the polyol pathway, inhibits glyceraldehyde triphosphate dehydrogenase  for its rapid regeneration (Bolaños et al., via increased superoxide anion generation (Ziegler et al., Most importantly, the proposed molecular events leading to neuronal oxidative stress in hyperglycemia, explicitly pars compacta and that may render them more sensitive to oxidative stress.Today, little doubt remains as to the hazard that a diabetic prelude represents in the development of PD. Even if the etiopathogenesis of PD remains largely enigmatic, in part due to our lack of comprehension regarding the seemingly selective vulnerability of the nigrostriatal dopaminergic pathway to undergo degeneration, ascending hypotheses grant a preeminent role to the existence of a particular phenotypic liability that characterizes neurons of the substantia nigra Hyperglycemia represents an additional cause of superoxide anion production, mitochondrial dysfunction, antioxidative bankruptcy, ubiquitin-proteasome system failure and dopamine toxicity that may well favor the early demise of nigrostriatal dopaminergic neurons. Thus, considering the deleterious effects of neuronal glucotoxicity, improving glycemic control in diabetic patients should represent a valuable strategy to decrease the risk of developing PD.DS wrote and edited the manuscript and drew the figure. JR wrote the manuscript. NS revised the manuscript. M-GM designed the manuscript, drew the figure and revised the manuscript.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.


Witches’ Broom Disease (WBD) affecting cacao plant and fruit in South and Central America. The severity and extent of this disease is prospected to impact the growing global chocolate market in a few decades. WBD is caused by the basidiomycete fungus Moniliophthora perniciosa. The methods used to contain the fungus mainly rely on chemical fungicides, such as copper-based compounds or azoles. Not only are these highly ineffective, but also their utilization is increasingly restricted by the cacao industry, in part because it promotes fungal resistance, in part related to consumers’ health concerns and environmental awareness. Therefore, the disease is being currently tentatively controlled through phytosanitary pruning, although the full removal of infected plant material is impossible and the fungus maintains persistent inoculum in the soil, or using an endophytic fungal parasite of Moniliophthora perniciosa which production is not sustainable. The growth of Moniliophthora perniciosa was reported as being antagonized in vitro by some yeasts, which suggests that they could be used as biological control agents, suppressing the fungus multiplication and containing its spread. Concurrently, some yeast-based products are used in the protection of fruits from postharvest fungal spoilage, and the extension of diverse food products shelf-life. These successful applications suggest that yeasts can be regarded a serious alternative also in the pre-harvest management of WBD and other fungal plant diseases. Yeasts’ GRAS  nature adds to their appropriateness for field application, not raising major ecological concerns as do the present more aggressive approaches. Importantly, mitigating WBD, in a sustainable manner, would predictably have a high socioeconomic impact, contributing to diminish poverty in the cacao-producing rural communities severely affected by the disease. This review discusses the importance/advantages and the challenges that such a strategy would have for WBD containment, and presents the available information on the molecular and cellular mechanisms underlying fungi antagonism by yeasts.Plant diseases caused by fungal pathogens are responsible for major crop losses worldwide, with a significant socio-economic impact on the life of millions of people who depend on agriculture-exclusive economy. This is the case of the  A great number of plant diseases are responsible for major crop losses with huge socio-economic impact, causing each year a worldwide estimated losses of 40 billion dollars . ParticuTo prevent plant diseases and protect crops from pests and pathogens, widely spread methodologies mainly correspond to applying chemical fungicides. The continued use of chemical fungicides leads to the development of fungicide resistance in the fungal pathogen  and, in Witches’ Broom Disease (WBD). This is caused by the basidiomycete fungus Moniliophthora perniciosa (Crinipellis perniciosa). The severity and extent of its manifestation is endangering the rapidly expanding and very quality-demanding chocolate market. This review focuses on this disease, the methodologies presently used for containment, and the innovative approaches that could be explored  biotrophic and (ii) saprotrophic. (i) The initial infection occurs in young meristematic tissues and susceptible actively growing tissues . The fungus penetrates through the stomatal openings, the bases of damaged trichomes and the husk of young fruits -respiration was associated with this transition  that bind to sulfur-containing proteins, (iii) targeting the pathogen DNA, this way inducing cell death, (iv) interrupting electron transport, this way causing the collapse of membrane potential, (v) promoting the generation of ROS, or (vi) interfering with nutrient uptake  inhibited in vitro the growth of several wood decay basidiomycetes including Serpula lacrymans, Postia placenta, Lentinus lepideus and Ophiostoma ulmi and phytopathogenic fungi, such as Rhizoctonia solani, Fusarium equiseti, Botrytis fabae, and Phytophthora infestans. Importantly, Torulaspora globosa against the phytopathogenic mold Colletotrichum graminicola, the causal agent of anthracnose disease in maize. All of these cases were reported as in vitro studies, none were performed in planta or in field. Otherwise, yeasts have often been proposed and used for the control of microbial contaminations at the postharvest phase    , Wickerhanomala) , Williopsaturnus , and Klu phaffii . Yet oths lactis , and by  acaciae  and Wingbertsiae . The bloo maydis . AlthougPichia guilliermondii antagonizes Botrytis cinerea   .Metschnikowia pulcherrima to compete with Botrytis cinerea, Alternaria alternata, and Penicillium expansum  has produced and developed a semi-commercial product named Tricovab CEPLAC4 . Additio CEPLAC4 . Two yeaackfruit . These o5. This is an expensive and drastic solution for plantations still operating, implicating a production lag phase that not all producers can afford. Moreover, more resistant varieties cannot avoid the incoming of more virulent fungal strains, which appears to be a world-wide trend (Witches’ Broom Disease of cacao, contributing to ease the associated severe social and economic implications to the life of millions of people.The cacao producers from Central and South America, where the WBD is a threat, are being encouraged to substitute their plantations with more resistant cacao plant varietiesde trend . The useThis is a research area with plenty to explore and a predictable great importance in the field of plant pathology in the next years.PF wrote the manuscript together with FC and CL. All authors read and approved the final manuscript.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.


Conducting electrophysiological measurements from human brain function provides a medium for sending commands and messages to the external world, as known as a brain–computer interface (BCI). In this study, we proposed a smart helmet which integrated the novel hygroscopic sponge electrodes and a combat helmet for BCI applications; with the smart helmet, soldiers can carry out extra tasks according to their intentions, i.e., through BCI techniques. There are several existing BCI methods which are distinct from each other; however, mutual issues exist regarding comfort and user acceptability when utilizing such BCI techniques in practical applications; one of the main challenges is the trade-off between using wet and dry electroencephalographic (EEG) electrodes. Recently, several dry EEG electrodes without the necessity of conductive gel have been developed for EEG data collection. Although the gel was claimed to be unnecessary, high contact impedance and low signal-to-noise ratio of dry EEG electrodes have turned out to be the main limitations. In this study, a smart helmet with novel hygroscopic sponge electrodes is developed and investigated for long-term usage of EEG data collection. The existing electrodes and EEG equipment regarding BCI applications were adopted to examine the proposed electrode. In the impedance test of a variety of electrodes, the sponge electrode showed performance averaging 118 kΩ, which was comparable with the best one among existing dry electrodes, which averaged 123 kΩ. The signals acquired from the sponge electrodes and the classic wet electrodes were analyzed with correlation analysis to study the effectiveness. The results indicated that the signals were similar to each other with an average correlation of 90.03% and 82.56% in two-second and ten-second temporal resolutions, respectively, and 97.18% in frequency responses. Furthermore, by applying the proposed differentiable power algorithm to the system, the average accuracy of 21 subjects can reach 91.11% in the steady-state visually evoked potential (SSVEP)-based BCI application regarding a simulated military mission. To sum up, the smart helmet is capable of assisting the soldiers to execute instructions with SSVEP-based BCI when their hands are not available and is a reliable piece of equipment for strategical applications. Conducting electroencephalographic (EEG) activity or other electrophysiological measures from human brain function might provide a nonmuscular medium for sending commands and messages to the external world, as known as a brain–computer interface (BCI). The purpose of BCIs is to convert human intentions into mechanical commands in order to control devices and machines without peripheral nervous systems and muscle movements. In recent years, a variety of BCI techniques have been developed and researchers have dedicated themselves to applying BCIs into real-world applications; for instance, motor imagery (MI) for stroke rehabilitation , steady-All aforementioned BCI methods exhibit several advantages and weaknesses compared with each other. For example, SSVEP shows a great signal-to-noise ratio  and relaIn recent decades, the developments of dry electrodes have been springing up in the fields regarding EEG measurements and BCI applications ,10. BiopIn this study, we proposed a smart helmet, which comes with semi-dry electrodes called hygroscopic sponge electrodes; the skin-contacting part is composed of sponge, which is hygroscopic and soft for the purpose of satisfying conductivity and comfort during EEG acquisition. Several properties of the electrode regarding EEG acquisition and BCI applications were investigated through the study, including the impedance of the electrodes and the correlation of the signals acquired via the classic wet electrodes and the proposed sponge electrodes. This novel electrode exhibits the advantages of both wet and dry electrodes: it is convenient to use as a wireless EEG accessory, and the impedance is lower than that of dry electrodes, yielding better signal quality. During the training for and execution of military missions, soldiers are usually equipped with combat helmets to protect their head; meanwhile, their hands constantly need to hold equipment, such as a rifle or a walkie-talkie. In this study, we proposed a smart helmet which integrated the novel hygroscopic sponge electrodes and a commercially available combat helmet for SSVEP-based BCI applications; with the smart helmet, soldiers can carry out additional instructions according to their intentions, i.e., through BCI techniques. As described above, wet electrodes require skin preparation and conduction gel to reduce the skin–electrode interface impedance . The desIn order to achieve lower impedance, some specifications are required for the sponge, including high water absorption, large contact area, and low resistance of the sponge material itself. Compared with polyurethane (PU) foam, pulp sponge features good electrical conductivity. There are two reasons for this. First, pulp sponge is composed of cellulose, which is a biomaterial with a hygroscopic nature. In addition to the porosity of the sponge structure, the hydrophilic cellulose molecule possesses higher water retention capacity. Second, the flexibility of wood-pulp fibers is larger than that of PU, so the soft sponge part can fit body surfaces perfectly, even in the areas covered with hair. Moreover, the electrode was designed to contact the skin via the dipped sponge, supported by a foam pad in order to avoid water effluence.In order to validate the performance of the developed electrode, an experiment of the impedance test was administered; the following sections elaborate the details.Ten healthy volunteer participants were recruited for the experiment . None of them had any history of neurological defects. The experiment was approved by the institutional review board (IRB) of National Chiao Tung University, Hsinchu, Taiwan.The experimental setup of the impedance test is shown in Four healthy subjects participated in the experiment . None of them had any history of neurological defects.The impedance test experiment of a variety of positions was performed to validate the performance of the proposed electrodes. Eight sponge electrodes were attached to wireless EEG equipment, Mindo BR8, developed by the Brain Research Center of National Chiao Tung University, Hsinchu, Taiwan ,25,26,27The EEG dynamics were acquired from sponge electrodes via Mindo BR8 and from disk electrodes (AgCl sintered) via Neuroscan simultaneously to validate the signal performance of the sponge electrode. In this experiment, eight channels of EEG signals were investigated, which acquired from one subject .The EEG activities were retrieved through the sponge electrodes that attached on the wireless Mindo BR8, and the channels we adopted were Fp1, Fp2, Fz, C3, C4, Pz, O1 and O2. The disk electrodes were placed beside the sponge electrodes to record the same region of EEG signal through Neuroscan a. The saAfter the impedance test and signal validation, another experiment would be implemented in order to validate the performance of the SSVEP-based BCI application utilizing the smart helmet. Furthermore, an algorithm called differentiable power was proposed to enhance the performance of the BCI system. In total, 21 healthy subjects participated in the experiment . None of them had any history of neurological defects.One of the most common methods to detect a target, i.e., flickering the object being focused upon, is using a power spectral density analysis (PSDA) . In receAn established BCI application based on SSVEP was demonstrated to further strengthen the effectiveness of the proposed smart helmet. The framework was adapted from , which iIn this study, four experiments were performed in order to validate the performance of the proposed smart helmet, including two impedance tests, a comparison between signals acquired from different electrodes, and an SSVEP-based BCI application. Besides the proposed smart helmet, two devices were adopted during the experiments. One was the well-known wet sensor EEG equipment: Neuroscan. The other was an eight-channel wireless EEG equipment called Mindo. The results of the experiments are elaborated as follows. Lower impedance indicates better conductivity between the scalp and the electrode; hence, two impedance tests regarding a variety of electrodes and scalp positions were investigated. Regarding the subjects, the impedance trends of all four electrodes is shown in Among all adopted electrodes, the spring-loaded metal electrode exhibited the highest impedance, and its impedance illustrated a fluctuating trend over time. Both the hygroscopic sponge electrode and the foam-based electrode exhibited lower impedance, and their impedance magnitudes showed a downward trend. In this part, we revealed the comparison between two signals, which were recorded from sponge electrodes via Mindo BR8 and disk electrodes (AgCl sintered) via the Neuroscan system. In We also performed the correlation analysis to power spectral density (PSD). The EEG data was processed by fast Fourier transform (FFT) to get PSD; The proposed smart helmet was applied to the subject during the SSVEP-based BCI application regarding the simulated military mission. The application consists of 30 trials; In this study, we developed a smart helmet and performed several experiments to investigate its characteristics and to validate its performance. The most important feature regarding the smart helmet is that it adopted the proposed hygroscopic electrodes. Impedance is considered as one of the most important properties regarding evaluating an electrode. Electrode impedance is expected to impact the signal-to-noise ratio during EEG recording . In the In the impedance test of a variety of electrodes, four kinds of electrodes were compared. However, the measurements of the electrodes were not bases on the exact same criteria because of the characteristics of each electrode. Two sides of the scalp were considered to administer the test; on the frontal side, the foam-based and the wet patch electrodes were placed because both these electrodes are not appropriate for application to hairy sites; on the occipital side, the spring-loaded metal electrode and the proposed hygroscopic electrode were placed to record the EEG dynamics. Another advantage of choosing frontal and occipital sides is that when performing EEG acquisition, all four electrodes were vertical to the ground, which minimized the effect of pressure caused by gravity. The results suggested that the impedance of the proposed electrode was comparable with the foam-based electrode, which exhibited the lowest impedance among three existing electrodes. Furthermore, unlike the foam-based electrode (Fp1), the proposed sponge electrode can be placed on sites with hair (O1).The impedance test of a variety of positions revealed the influence of different channels regarding EEG acquisition. Due to the natural variabilities of humans and their skin conditions, such as skin thickness, the degree of skin hydration, and the number of hair follicles ,32, the The smart helmet with the attached hygroscopic sponge electrodes, which feature convenience and low impedance, was developed for EEG acquisition, especially for BCI applications. It possesses the characteristics of comfort, good accessibility, and high fidelity of signals because of the design, the materials, and the structure. Comparing the sponge electrode with existing dry contact electrodes, it is not only applicable for hairy sites on the scalp, but also exhibits lower impedance, which enhances the feasibility and performance regarding BCI applications. It also strengthens the effectiveness of the smart helmet with the low skin–electrode interface impedance and high correlation between the signals acquired from the proposed electrode and the classic Ag/AgCl electrode. To sum up, the smart helmet is capable of assisting soldiers to execute extra tasks with BCI techniques and is a reliable piece of equipment for strategical applications.


The advent of cell-cell and cell-extracellular adhesion enabled cells to interact in a coherent manner, forming larger structures and giving rise to the development of tissues, organs and complex multicellular life forms. The development of such organisms required tight regulation of dynamic adhesive structures by signaling pathways that coordinate cell attachment. Integrin-mediated adhesion to the extracellular matrix provides cells with support, survival signals and context-dependent cues that enable cells to run different cellular programs. One mysterious aspect of the process is how hundreds of proteins assemble seemingly spontaneously onto the activated integrin. An emerging concept is that adhesion assembly is regulated by autoinhibition of key proteins, a highly dynamic event that is modulated by a variety of signaling events. By enabling precise control of the activation state of proteins, autoinhibition enables localization of inactive proteins and the formation of pre-complexes. In response to the correct signals, these proteins become active and interact with other proteins, ultimately leading to development of cell-matrix junctions. Autoinhibition of key components of such adhesion complexes—including core components integrin, talin, vinculin, and FAK and important peripheral regulators such as RIAM, Src, and DLC1—leads to a view that the majority of proteins involved in complex assembly might be regulated by intramolecular interactions. Autoinhibition is relieved via multiple different signals including post-translation modification and proteolysis. More recently, mechanical forces have been shown to stabilize and increase the lifetimes of active conformations, identifying autoinhibition as a means of encoding mechanosensitivity. The complexity and scope for nuanced adhesion dynamics facilitated via autoinhibition provides numerous points of regulation. In this review, we discuss what is known about this mode of regulation and how it leads to rapid and tightly controlled assembly and disassembly of cell-matrix adhesion. The emergence of life is an enigmatic question that humankind has pondered for millennia. At some point millions of years ago, the last universal common ancestor (LUCA) appeared and all organisms on earth descended from this initial cell  to form cell-matrix junctions. The ability of cells to form sheets of cells and attach to an underlying matrix was central to the development of multicellular animal life.These attachment points also developed into sensitive sensory modules, able to feel the mechanics of the microenvironment and adopt the role of mechanotransduction centers—enabling cells to monitor and respond to mechanical cues and convert them into biological signals to elicit different cellular responses.Key to the development of complexity is the ability to regulate adhesion and control the proteins that assemble together to form adhesive structures. One way that proteins can be dynamically regulated is via formation of an intramolecular interaction that maintains the protein in an inactive state until adhesion assembly is required. The regulation of autoinhibition, and the factors that enable regulation of the activity of the protein, provides regulatable checkpoints in the system .The concept of autoinhibitory domains can be found throughout biology—these are regions of proteins which can form intramolecular interactions which regulate behavior and activity. We refer the reader to the review on autoinhibitory domains by Pufall and Graves as it provides an excellent, still highly relevant description of the fundamentals of autoinhibition  can relieve this autoinhibitory interaction and drive tail separation, propagating a conformational rearrangement of the ectodomains and unfurling to reveal the ligand-binding motifs  in fibroblasts -like region in F3—this domain is termed integrin binding site 1 (IBS1). As mentioned in the previous section, the interaction between the talin head domain with integrin is sufficient to relieve autoinhibition of the integrin, exposing the previously cryptic binding sites for integrin ligands. However, talin itself is regulated by multiple layers of autoinhibition.The first evidence that talin was autoinhibited came from the initial structural and biochemical characterization of talin in 1987  of R13 limits actin binding  RIAM is activated, which translocates talin to the plasma membrane where (ii) the talin head-tail autoinhibition is relieved. (iii) Once active, talin relieves the autoinhibition of integrin. (iv) By connecting integrins to the actin retrograde flow, talin autoinhibition is further relieved, first with activation of enhanced actin binding to ABS3. (v) As force increases, mechanical activation of vinculin-binding sites in talin occurs, initially by unfolding of the R3 domain, the least stable of the rod domains  to which Vd1 can bind  are well-characterized and, being a kinase, a striking example of coupling regulation of protein activity to downstream signaling cascades. FAK is another linear multidomain protein with numerous binding sites for ligands. As the name suggests, FAK contains an enzymatically active kinase domain able to phosphorylate tyrosine residues in target proteins. FAK comprises an N-terminal FERM domain, a kinase domain, a ligand-binding region and a C-terminal 4-helix bundle termed the focal adhesion targeting (FAT) domain .At rest, FAK adopts an autoinhibited, closed conformation where the FERM domain directly binds and occludes both the catalytic cleft and the activation loop of the kinase domain  and a mutation in the kinase domain (Phe596Asp) .An important regulator of the integrin adhesion complex assembly is Rap1-interacting adapter molecule (RIAM). RIAM is a Rap1 effector that can interact directly with talin and translocate it to the plasma membrane, and thus into close proximity to the integrins  . This INThis Rap1:RIAM:talin nexus is thus tightly controlled in a myriad of ways, and the dynamic balance of activity status of each protein is implicitly entangled. RIAM is likely activated by active Rap1 Bos, , and theRelief of autoinhibition of FAK following activation at adhesion sites leads to FAK autophosphorylation of Tyr397 Frisch,  which prSrc recruitment to, and phosphorylation of, downstream molecules allows many pathways to be regulated through Src, and drives crosstalk between integrin, Src and Rho-family GTPases  (One common mechanism for relieving autoinhibition is s (PTMs) . Here, as (PTMs) , sumoylas (PTMs) , etc. Thproteolytic cleavage  led to insight into the loss of essential developmental phenotypes  and, onto this hub, hundreds of proteins are recruited , will trigger changes to the system as it reorients to this new homeostasis, adopting an altered adhesion signaling complex which augments the programming of the cell.BG and RK wrote the manuscript.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.