Hosts must recognize pathogen invasion and respond rapidly. The need for speed requires a low threshold for triggering a response, causing occasional false alarms. Host immune systems therefore require strong negative regulators to shut down unnecessary responses. The evolutionary consequences of rapid trigger dynamics balanced by negative regulators have received little attention. Here, we emphasize four aspects that influence the evolutionary genetics of immunity: diverse gene families of rapidly acting triggers opposed by slower-acting negative regulators, pathogen attack against negative regulators, diversifying selection of negative regulators by pathogen pressure, and heritability of immune-related disease from imbalance between triggers and negative regulators.Acropora digitifera has around 500 genes, sea urchins have approximately 200, and zebrafish have about 385 [A broad sensor array of receptors, located on the cell surface or in the cytosol, recognize pathogen-associated molecular patterns and trigger innate immunity. Common triggers include the peptidoglycan recognition proteins, gram-negative binding proteins, Toll-like receptors (TLRs), nucleotide-binding domain leucin-rich repeats (NLRs), and the cyclic GMP-AMP synthase (cGAS) system that detects double-stranded DNA (dsDNA) , 2. In tbout 385 . SpongesThese receptors trigger immune cascades that are often evolutionarily ancient within groups or across broad domains of life. Key components of such cascades include nuclear factor-κB (NF-κB) transcription factors, interferons (IFNs), interleukins, and Toll and immune deficiency (IMD) pathways –9. The pThe need for speed in immune response  demands All organisms have an impressive array of negative regulators. Post-transcriptional modification of innate sensors and downstream molecules suppress the effect of the pattern recognition receptor triggers , 14. TriDifferent organisms will tune the tradeoff between faster response and lower error rates in different ways. A focus on the tuning of that tradeoff may lead to interesting comparative insights about the design of immune regulation.Leishmania exploits a negative TLR regulator to suppress production of key proinflammatory cytokines [Attackers use diverse mechanisms to block the expression of host innate immunity , 25–27. ytokines , virusesytokines , and manytokines .Invaders of insects alter the expression of an impressive variety of miRNAs, including negative regulators. Some pathogens express their own miRNAs to alter host immunity. The function of most miRNAs remains unknown, providing a rich topic for further study .Tumors also evolve to down-regulate immunity against cancer cells and often manipulate the normal repressors of immunity. For example, Vinay and colleagues  note thaHosts gain by altering the specific sequences of their negative regulators to avoid recognition or mimicking by pathogen effectors. Negative immune regulators that are under attack may form diverse gene families and evolve relatively rapidly. Few prior studies have focused explicitly on the diversity and evolutionary rate of negative regulators.Drosophila, Lee and Ferrandon [Drosophila, genes involved in the response signaling cascade tended to be more conserved than recognition proteins, but those that functioned primarily in modulating the immune response showed evidence for positive selection far above the genomic average [In errandon  note tha average .The nature of diversifying selection depends on the specificity of mechanisms by which attackers manipulate host immunity. In human immunity, attack against negative immune regulators may be relatively nonspecific, in the sense that the attackers may not be differentially targeting molecules based on sequence variability. By contrast, for the small RNAs that regulate immunity in plants, insects, and other organisms, the RNA sequences and the mechanisms of manipulation are likely to be highly specific. The more sequence-specific the mechanisms that attackers use to manipulate negative immune regulators, the stronger the diversifying selection will be.Regulation by strongly opposed triggers and repressors is prone to failure ( discussePlants may use miRNAs to tune the threshold for the triggering of immunity by the NLR pattern recognition receptors. Canto-Pastor and colleagues  found thSimilarly, constitutively expressed inhibitors of the type I IFN response in human immunity may “determine a threshold of activation of an antiviral response” . A quantGenetic variability in triggers and negative regulators can lead to mismatches in the opposing regulatory components of innate immunity and various types of immune-related disease. One possibility concerns the various immune-related disorders associated with misexpression of type I IFN . AnotherOur hypothesis and these preliminary examples suggest that further study of genetic variability in the positive and negative regulators of immune cascades may lead to interesting insights about the heritability of immune misregulation.


Social capital is essential for healthy living in later life, and evidence indicates the importance of neighborhood context in facilitating access to this resource. However, additional research is needed to compare how objective and subjective indicators of context shape this association. We examine links between objective (census tract-level population density and composition) and subjective (perceived safety and quiet) indicators of neighborhoods and social capital . Data include participants age 65+ (N=259) from Wave 3 (2015) of the longitudinal Social Relations Study based in Detroit, Michigan. Multivariate analyses indicate respondents living in more densely populated neighborhoods reported more frequent contact with network members. Those perceiving their neighborhood as safe reported larger networks, more trust in and greater support exchanged among neighbors. Findings suggest both objective and subjective indicators of neighborhood context shape access to social capital, but perceptions play a larger role.


OBJECTIVES/SPECIFIC AIMS: The objective of this project is the application of complex fusion models, which combine observed and modeled data, to areas with sparse monitoring networks with multiple chemical components is under-developed. Such models could provide improved accuracy and coverage for air quality measurement predictions, an area greatly limited by the amount of missing data. METHODS/STUDY POPULATION: This project focuses on the development of methods for improved estimation of pollutant concentrations when only sparse monitor networks are found. Sparse monitoring networks are defined as areas where fewer than three criteria air pollutants (based on EPA standards) are monitored. Particularly, a multivariate air pollutant statistical model to predict spatio-temporally resolved concentration fields for multiple pollutants simultaneously is developed and evaluated. The multivariate predictions allow monitored pollutants to inform the prediction of nonmonitored pollutants in sparse networks. RESULTS/ANTICIPATED RESULTS: Daily, ZIP code level pollutant concentration estimates will be provided for 8 pollutants across South Carolina, and goodness of fit metrics for model variants and previously established methods will be compared. DISCUSSION/SIGNIFICANCE OF IMPACT: These methods utilize only widely available data resources, meaning that the improved predictive accuracy of sparsely monitored pollutant concentrations can benefit future studies in any US area by improving estimation of health effects and saving resources needed for supplemental air pollutant monitoring campaigns. Our method for estimation attempts to improve predictive accuracy and data availability for sparsely monitored pollutants and areas.


A 78-year old male presented to the emergency department after accidental dislodgement of his chronic gastrostomy tube. A replacement gastrostomy tube was passed easily through the existing stoma and flushed without difficulty. Confirmatory abdominal radiography demonstrated contrast in the proximal small bowel, but the patient subsequently developed epigastric pain and refractory vomiting. Computed tomography revealed the tip of the gastrostomy tube terminating in the pylorus or proximal duodenum. This case highlights gastric outlet obstruction complicating the replacement of a gastrostomy tube and the associated radiographic findings. A 78-year-old male with a history of stroke presented to the emergency department after accidental dislodgement of his chronic gastrostomy tube approximately five hours prior. The patient offered no other complaints and denied abdominal pain, nausea, or vomiting. On examination, he appeared comfortable with unremarkable vital signs. His abdomen was non-tender and demonstrated a patent, mature gastrostomy stoma. A replacement gastrostomy tube was passed easily through the existing stoma and flushed without difficulty. Confirmatory abdominal radiography revealed contrast in the duodenum and proximal jejunum, but no portion of the stomach was outlined . ShortlyGastric outlet obstruction related to gastrostomy tubes is rare.4What do we already know about this clinical entity?Dislodged gastrostomy tubes are commonly repositioned or replaced in the emergency department. Gastric outlet obstruction is a potential, albeit rare, complication.What is the major impact of the image(s)?Emergency physicians should be familiar with the appearance of gastric outlet obstruction due to gastrostomy tube malposition on contrast-enhanced plain radiography.How might this improve emergency medicine practice?Early identification of gastric outlet obstruction on plain radiography can prevent patient discomfort and the need for additional advanced imaging.


Notopteridae  represents an old fish lineage with ten currently recognized species distributed in African and Southeastern Asian rivers. Their karyotype structures and diploid numbers remained conserved over long evolutionary periods, since African and Asian lineages diverged approximately 120 Mya. However, a significant genetic diversity was already identified for these species using molecular data. Thus, why the evolutionary relationships within Notopteridae are so diverse at the genomic level but so conserved in terms of their karyotypes? In an attempt to develop a more comprehensive picture of the karyotype and genome evolution in Notopteridae, we performed comparative genomic hybridization (CGH) and cross-species (Zoo-FISH) whole chromosome painting experiments to explore chromosome-scale intergenomic divergence among seven notopterid species, collected in different African and Southeast Asian river basins. CGH demonstrated an advanced stage of sequence divergence among the species and Zoo-FISH experiments showed diffuse and limited homology on inter-generic level, showing a temporal reduction of evolutionarily conserved syntenic regions. The sharing of a conserved chromosomal region revealed by Zoo-FISH in these species provides perspectives that several other homologous syntenic regions have remained conserved among their genomes despite long temporal isolation. In summary, Notopteridae is an interesting model for tracking the chromosome evolution as it is (i) ancestral vertebrate group with Gondwanan distribution and (ii) an example of animal group exhibiting karyotype stasis. The present study brings new insights into degree of genome divergence vs. conservation at chromosomal and sub-chromosomal level in representative sampling of this group. This ancestral teleost lineage retained primitive anatomical features 4 and, considering their very ancient origin (200 Mya), their current distribution pattern could reflect the vicariance events occurring after the Gondwana’s break-up6. While some osteoglossiform families  are restricted to Africa, other ones  exhibit a patchy distribution, with species endemic to different continents7.The monophyletic fish order Osteoglossiformes represents an ancient teleost group with a geographic distribution restricted to the freshwater river basins13]. This lack of data is probably due to the worldwide distribution of the whole group and, on the other hand, the endemic status of majority of its species, thus hindering integrative studies that allow a globalized view of its evolutionary process.Their wide geographic distribution and basal position in the teleost phylogeny, qualifies this group as an excellent model for systematic, genomic, cytogenetic and evolutionary studies. However, only few cytogenetic and genomic studies have been undertaken so far, covering a limited number of osteoglossiform species , and CGH homology (intraclade genomic similarity). The groups/species relationships  followed phylogenetic hypothesis based on DNA molecular markers available for family


We highlight a range of strategies for establishment of three dimensional (3-D) airway and intestinal organoid models and evaluate the limitations and potential improvements in each system, focusing on their application in CF. The in vitro CFTR functional assays in patient-derived organoids allow for preclinical pharmacotherapy screening to identify responsive patients. It is likely that organoids will be an invaluable preclinical tool to unravel disease mechanisms, design novel treatments, and enable clinicians to provide personalized management for patients with CF.Cystic fibrosis (CF) is an inherited disorder where individual disease etiology and response to therapeutic intervention is impacted by CF transmembrane regulator (CFTR) mutations and other genetic modifiers. CFTR regulates multiple mechanisms in a diverse range of epithelial tissues. In this Review, we consolidate the latest updates in the development of primary epithelial cellular model systems relevant for CF. We discuss conventional two-dimensional (2-D) airway epithelial cell cultures, the backbone of  It is cMajor advances in symptom management have been instrumental in delaying disease progression in CF. Administration of enzyme replacement therapy, nutritional support, mucolytic agents airway clearance techniques, and antibiotics for bacterial lung infection, along with early detection of disease  has dramatically improved life expectancy in CF patients over the last 4 decades. The median age of death is now above 40 years  airway and rectal organoids. We also discuss their emerging use as More than 2,000 CFTR mutations have been identified so far and at least 336 of these are reported to lead to symptoms characteristic of CF , reduced episodes of pulmonary exacerbations and improved body mass index (BMI)   restored F508del-CFTR channel activity to 15% of wild-type CFTR in in vitro  epithelial cell lines. It is thus not surprising that this model has a higher propensity of false-positive and false-negative “hits” compared to that performed in primary human bronchial epithelial (HBE) cells.One of the major hurdles to the development of novel treatment regimens in CF is the bench-to-bedside translation of scientific knowledge. Many drugs that perform well in a laboratory setting fail to advance in clinical trials, largely due to inappropriate selection of Although CF is a monogenic disease, the diverse mutation variants identified within the CFTR gene as well as the presence of modifier genes (known and unknown), warrants adoption of new technologies to extend research capabilities. There is a clear unmet need for a representative library of patient-specific epithelial cell models for disease modeling, preclinical testing of drug response, and biobanking for future drug discovery. The cell models can be derived from embryonic stem cells (ESCs), induced pluripotent stem cells (iPSCs), or tissue-resident adult stem cells. Whereas, ethical concerns pertaining the source of human ESC, limit their use in research, generation of CFTR gene corrected iPSC lines enable disease modeling, drug discovery and toxicology studies [reviewed in  fluid and induced sputum samples usually do not provide sufficient cell counts to initiate culture. Therefore, supply of CF patient-derived HBE cells are often limited and hard to come by.The pulmonary epithelium is divided into three regions; upper , lower (trachea and primary bronchi), and distal small airway epithelia . Persistent inflammation, bacterial colonization, and airway structural changes in CF occur in the lower respiratory tract. Primary HBE cells are therefore the gold standard for studying CF disease pathogenesis and evaluating CFTR functional response  cells are increasingly used as surrogates for the lower airway epithelium in CF research  values . Expansion of epithelial cells is often necessary in the initial passages for biobanking purposes and to generate enough cell numbers needed to differentiate cells under air-liquid interface (ALI) conditions. HBE cells are most “− transport in these cells is also preserved.Conditionally reprogrammed cells (CRCs) are generated by co-culturing patient-derived airway epithelial cells with irradiated fibroblast feeder cells. Specialized conditioned media (termed F-Media) which contains Rho-associated protein kinase (ROCK) inhibitor, promotes serial passages of airway epithelial cells and enhances population doubling without compromising the characteristic epithelial cell morphology. Both ROCK inhibitor and the feeder layer are essential in maintaining the stem cell-like phenotype evident in CRCs  demonstrated some advantage over the standard CRCs. The modified CRCs could support airway epithelial cell growth up to passage 10 with robust formation of pseudostratified epithelium at the extended passage, although a modest decrease in CFTR-dependent Cl+ currents and Cl− conductance declined fairly rapidly after serial passage 6  and bone morphogenic protein (BMP), showed airway epithelial cells could be expanded up to 18–25 passages without loss of proliferative potential  or are cultured in such a manner that they develop ECM-like scaffolds between them, thus mimicking the in vivo phenotype more faithfully. While cell-derived ECMs such as Matrigel have been most commonly used for development of organoids, their undefined components introduce inconsistency in replicating the native 3-D culture environment , which establishes intercellular cues or network in the Different methods for generating organoids (or spheroids) from human lung cells have been described to date, each producing organoids with distinct definitive structure and cellular compositions. Barkauskas and colleagues have elegantly reviewed the different lung organoids (airway organoids inclusive) established from varying lung progenitor cell populations, including basal cells in the proximal airways, secretory club Clara cells in bronchioles and alveolar type II cells in the alveoli, as well as those from embryonic and iPSCs , patient-derived intestinal epithelial cells also present an invaluable resource in characterizing the relationship between the CFTR gene mutation and the disease phenotype. More recently, they play an important role in the development of HTS strategies to elucidate novel drugs for the treatment of CF.ex vivo conditions. Other limitations to the use of rectal biopsies include the small number of biopsies collected (4–8 biopsies) and the need for all biopsies to be tested on collection day i.e., the biopsies cannot be preserved for further testing.Compared to human airway tissue, colon tissue damage in CF patients is minimal and the rectal epithelium is accessible in a less invasive manner. The abundant expression of CFTR in the distal colon makes rectal biopsies an attractive cellular source for interrogating CFTR function , inhibitors of TGF-β and BMP signaling and the basement membrane matrix (matrigel). Intestinal organoids can be indefinitely cultured and remain genetically and phenotypically stable upon repeated passaging and long term culture  overcome these limitations by extending the use of rectal biopsies in cultures. Intestinal organoids can be grown from crypts isolated from freshly isolated rectal biopsies. Crypts are rich in Lgr5The primary functional assay to assess CFTR activity in organoids, the forskolin-induced swelling (FIS) assay is CFTR dependent. This approach does not measure the net transepithelial ionic transport. Rather, forskolin is used to stimulate intracellular cAMP production which then activates CFTR at the plasma membrane. CFTR activation drives chloride and fluid flux to the organoid lumen , causing rapid swelling of organoids with functional CFTR or those derived from non-CF subjects. This swelling effect is significantly reduced or absent in organoids derived from CF subjects who exhibit differing rates of FIS with different classes of CFTR mutations and also between individuals with the same CFTR mutation , are either absent or present in negligible amounts in the gut to be functional (Rajendran et al., SAW, NTA, and SLW wrote the manuscript with critical input from CKH, LKF, AK and AJ.AK is a scientific advisor for Unizyme Laboratories A/S. The remaining 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.


Acute traumatic patellar dislocations are encountered with relative frequency, making up 3% of all knee injuries. Typically witnessed in younger patients following sporting injuries, this injury can be debilitating, potentially leading to recurrent dislocation, pain, reduction in activity and patellofemoral osteoarthritis.Management of this injury remains controversial, and as such detailed magnetic resonance imaging (MRI) is increasingly recommended to help illustrate the exact nature of osteochondral and soft tissue injury, with a view to assessing the anatomical sequelae of patellar dislocation as well as the potential of recurrence and dictating the need for either conservative or surgical management in the acute setting. As such, awareness of the typical MRI findings in traumatic patellar dislocations may potentially aid in pursuing appropriate intervention for this pathology.This case describes a 33-year-old gentleman presenting to the emergency department following patellar dislocation. After failed departmental closed reduction, this patient progressed on to definitive anatomical MRI assessment followed by acute surgical intervention in the form of medial patellofemoral ligament (MPFL) repair. This case allows for both illustration and discussion of typical radiological features associated with traumatic patellar dislocation. Acute traumatic patellar dislocations are encountered with relative frequency, making up 3% of all knee injuries . TypicalAssessment of a primary traumatic patellar dislocation is multifaceted, involving initial clinical history and examination followed by initial plain radiographic evaluation involving anteroposterior, lateral and axial views . While pThe following case report allows for illustration and discussion of some of the typical MRI findings one may encounter following acute patellar dislocation that may potentially precipitate subsequent definitive surgical intervention.A 33-year-old gentleman presented to the emergency department describing substantial right knee pain and swelling following a rotational injury. Inspection indicated significant joint effusion and subsequent examination showed absence of straight leg raise on the affected side. Plain radiographic evaluation of the knee showed lateral displacement of the patella Figure  confirmiRadiological investigations displayed evidence of lateral patellar dislocation on plain radiograph Figure , as wellIn the context of failed closed reduction as well as confirmation of the diagnosis, associated soft-tissue injuries and presence of risk factors for recurrent dislocation, this patient subsequently proceeded to definitive surgical treatment in the form of MPFL repair, with excellent postoperative function and no further dislocation at 46 months postoperatively.Acute patellar dislocations are debilitating, with up to 58% of patients noting persistent limitation to strenuous activity and 55% failing to return to sport at six months following index injury ,4. WhileDetailed anatomical illustration of the patellofemoral joint and surrounding structures at time of injury has thus become increasingly important aspect of initial evaluation following this injury -8. This Given both the increased diagnostic role of MRI and the relative frequency in which this injury is encountered, particularly in the adult population, awareness of both the plain radiographic features and more importantly the MRI findings associated with acute traumatic patellar dislocation as discussed above may aid in improved detection, earlier management and prevention of recurrent dislocations, hence reducing the potential debilitating effect of this injury ,2.Traumatic patellar dislocations most commonly occur laterally and are associated with injury to both the medial patellofemoral ligament and medial patellar retinaculum. MRI is becoming an increasingly important diagnostic tool in assessment of traumatic patellar dislocations, useful in both detecting associated cartilage and soft tissue disruption as well as risk factors for recurrent dislocation. As such, awareness of the typical MRI findings in traumatic patellar dislocations outlined in the above case may potentially aid in pursuing either definitive conservative or surgical treatment in this pathology.


Innate immune cells trigger sophisticated intracellular signaling pathways via innate immune receptors, including membrane-bound or cytosolic receptors3. Host innate immune activation results in the production of multiple effector molecules, including cytokines and chemokines as well as antimicrobial proteins, to combat invading pathogens and parasites4. In addition, innate immune cells, such as macrophages and dendritic cells, are the principal antigen-presenting cells that activate T cells, which are involved in more specific and delicate immune responses6. In particular, Th1 cell-mediated interferon (IFN)-γ can efficiently promote cell-autonomous host defenses against intracellular parasitic and mycobacterial infections through various effectors, including IFN-inducible GTPases, inducible nitric oxide synthase, and autophagy proteins8.Host–pathogen interaction is considered a highly dynamic process between diverse microbial pathogens and hosts in all stages of pathogenic infection, from invasion to dissemination. Upon pathogenic infection, innate immune systems respond to pathogen-associated molecular patterns and activate immediate host inflammatory and antimicrobial responsesHere, a special issue of articles presents recent findings on how host immune and pathological responses are generated by intracellular signaling machinery, which can be activated at various stages of host–pathogen interaction, and further describes how pathogens subvert host protective responses to establish an infection in the harsh environment inside host cells. In addition, this special issue includes invited reviews that discuss current knowledge of host defensive components and pathways, i.e., immune receptors, cytokines, signaling molecules, autophagy, and the microbiota. We further describe recent innovative trials based on studies of the host–pathogen interface involving the therapeutic utilization of bacteria in the context of anticancer treatments. A comprehensive understanding of host–pathogen interaction will provide new insights into the identification of novel targets for both host effectors and microbial factors and will lead to new therapeutic treatments for infections and other human diseases.9. Lee and coworkers provide a comprehensive summary of viral strategies that evade host cytosolic sensing to facilitate intracellular infection and replication of viruses. Although innate immune responses are essential for combating viral infections, they should be tightly regulated to prevent harmful host responses. Lee and coworkers also review recent findings on the positive and negative regulatory mechanisms for intracellular sensors in-host cells. Recent studies have uncovered key signaling molecules that participate in the recognition of cytosolic pathogen-associated molecular patterns (PAMPs) and the activation of innate immune responses against viral infection11. Ahn and Barber highlight the immune regulatory functions of stimulators of interferon genes and cyclic dinucleotides that can recognize microbial DNA from various pathogens, including bacteria, viruses, and parasites, as well as cytosolic DNA of self-origin from host cells. Investigations on the abundance of in-host effector mechanisms and pathogenic strategies will provide an ideal opportunity to develop innovative tools that can be utilized to modulate microbial pathogenesis.When encountering the host defense system, numerous intracellular pathogens employ a variety of evolved strategies to escape, modulate, and hijack host immunity during infection14. Jo and coworkers summarize the function of autophagy and xenophagy in mycobacterial infection and how to manipulate autophagy pathways to promote host-targeted therapeutics against mycobacterial infection. A more comprehensive understanding of the elaborate mechanisms by which host autophagy pathways overcome bacterial pathogenesis strategies may lead to more efficient antimicrobial defense strategies against numerous intracellular pathogens that can reside and replicate within the host cell cytoplasm16.Autophagy, a cell-autonomous catabolic pathway involving lysosomal degradation of cargos, is becoming recognized as an innate effector mechanism to enhance pathogen control and resolution of tissue pathology associated with infection18. Recent studies have also revealed the function of cytokines, including type I IFNs, IL-15, and IL-18, in the bystander activation of CD8 + T cells, which are responsible for tissue injury in viral infections20. Shin and Kim will present a new concept of bystander response, which leads to pathological or protective outcomes, depending on the context. An understanding of the IFN-inducible responses in various aspects of host–pathogen interactions will be essential for the development of effective therapeutics that also minimize tissue damage.When considering the role of type I and II interferons (IFNs) in the activation of antiviral and Th1 immune responses, respectively, it is generally accepted that IFN actions are typically required for protective immune responses against viruses and intracellular bacteria. Yamamoto and Sasai discuss recent progress on the cell-intrinsic defense mechanisms by which IFN-inducible GTPase-dependent host defenses exhibit antiparasitic and antibacterial responses. IFN-γ-inducible GTPases, including guanylate binding proteins (GBPs) and immunity-related GTPases (IRGs), are essential in various aspects of immune protective functions, including the release of bacterial components, production of reactive nitrogen species, inflammasome activation, and autophagy21. To date, the majority of studies on host–pathogen interactions have focused on comprehensive elucidation of the innate and adaptive immune responses during infection. Recently, several efforts have been made to manipulate genetically modified bacteria for utilization as a new form of therapeutic agent for the treatment of cancer22. Min and coworkers review the current trials, perspectives, and limitations of therapeutic approaches using live bacteria with tumor-targeting ability and discuss recent advances in our understanding of bacterium-tumor cell interactions.Accumulating evidence supports the importance of the gut microbiota in human health and diseases. Takeda and Maeda summarize the recent findings of molecular details in-host-microbe interactions in rheumatoid arthritis and how alterations at the interface between the host and microbes result in excessive inflammation and infection. In addition, this review will introduce multiple metabolic and immunological mechanisms by which the gut and oral microbiota induce the development of arthritisIn revealing a vast array of strategies adopted by host cells and pathogens during various stages of infection, paradigms are shifting. Accumulating evidence indicates a more diverse impact of pathogenic manipulations of host cells than previously known. This special issue highlights recent advances in studies that extend traditional concepts of host–pathogen interactions toward new insights on innate and adaptive immune signaling, molecular pathogenesis, host-directed therapy, and manipulation of bacteria for anticancer therapy. We believe that the cutting-edge reviews presented here will provide novel insights into the growing area of host–pathogen interface signaling, thereby expanding basic knowledge for use in clinical applications. Studying the basic and applied fields relating to this topic will promote our knowledge of highly complex host–pathogen relationships and facilitate future investigations in this area for the development of improved therapies against infections.


PARKIN gene cause early-onset Parkinson’s disease (PD). Despite the high proportion of still missing phenotyping data in the literature devoted to early-onset PD, studies suggest that, as compared with late-onset PD, PARKIN patients show dystonia at onset and extremely dose-sensitive levodopa-induced dyskinesia (LID). What pathophysiological mechanisms underpin such early and atypical dyskinesia in patients with PARKIN mutations? Though the precise mechanisms underlying dystonia and LID are still unclear, evidence suggests that hyperkinetic disorders in PD are a behavioral expression of maladaptive functional and morphological changes at corticostriatal synapses induced by long-term dopamine (DA) depletion. However, since the dyskinesia in PARKIN patients can also be present at onset, other mechanisms beside the well-established DA depletion may play a role in the development of dyskinesia in these patients. Because cortical and striatal neurons express parkin protein, and parkin modulates the function of ionotropic glutamatergic receptors (iGluRs), an intriguing explanation may rest on the potential role of parkin in directly controlling the glutamatergic corticostriatal synapse transmission. We discuss the novel theory that loss of parkin function can dysregulate transmission at the corticostriatal synapses where they cause early maladaptive changes that co-occur with the changes stemming from DA loss. This hypothesis suggests an early striatal synaptopathy; it could lay the groundwork for pharmacological treatment of dyskinesias and LID in patients with PARKIN mutations.Mutations in the  PARKIN gene cause autosomal recessive juvenile parkinsonism (ARJP) (PARKIN mutations differ from those observed in idiopathic Parkinson’s disease (iPD). Unlike iPD, neuronal loss is greater in the substantia nigra pars compacta (SNpc) than in the locus coeruleus in most parkin cases. Also, Lewy bodies are not detected in the majority of parkin cases, suggesting a difference in the pathogenic processes between PARKIN-related disease and iPD  . Autopsy and iPD . Also, Pfeatures , 2018. Efeatures . Early d and DJ1 . Since PPARKIN mutations and the recent evidence that parkin modulates glutamatergic neurotransmission. We discuss the hypothesis that early hyperkinetic symptoms of PD patients with PARKIN mutation may be considered manifestations of corticostriatal synaptopathy. These features can be linked to dopamine depletion, a well-recognized mechanism in the literature, but they also may be related to a potential direct effect of parkin on the corticostriatal synapse.Here, we review the clinical evidence for early hyperkinetic symptoms in patients with PARKIN gene in two families , prompts screening for PARKIN mutations along with GTP cyclohydrolase 1 and tyrosine hydroxylase in clinically diagnosed DRD patients  . MSNs arThe molecular signatures that have recently emerged as key signals in LIDs are ERK1/2 phosphorylation , abnormaPARKIN patients. Clinical studies with [18F] Dopa as a marker of the functional integrity of pre-synaptic dopamine nerve terminals have shown that Parkin mutated patients have widespread pre-synaptic dopaminergic deficits  and thus regulates its function: loss of parkin increases KAR levels in the human cortex and KAR function in glutamatergic hippocampal neurons in vitro (in vitro study showing that parkin-deficient hippocampal neurons exhibit significantly reduced AMPA receptor-mediated currents (PARKIN mutations disturb glutamatergic synaptic transmission by directly changing NMDAR, AMPAR, and KAR receptor function. Since parkin is widely expressed in various brain regions and neuron types, including cortical neurons and MSNs, (in vitro . At the in vitro . The rolcurrents . The sugcurrents . Hence, nd MSNs,  it is coParkin gene. Unfortunately, studies on the nigrostriatal pathway revealed many discrepancies probably deriving from different experimental conditions and/or from in vivo age-dependent compensatory mechanisms. Increased spontaneous DA release from SNc DA neurons was found in Parkin-knockout mice aged 8 to 9 months (Parkin knockout mice (Parkin-knockout mice, DA levels were reported to be indistinguishable (Parkin-knockout mice (Parkin-knockout mice. Studies on corticostriatal synapses performed by intracellular recordings of MSNs revealed impairment in long-term depression and long-term potentiation in Parkin-knockout mice (Parkin-knockout mice do not show DA neuron loss, motor impairment, dystonic phenotype or hypersensitivity to levodopa (PARKIN mutations were observed to potentiate glutamatergic transmission by modifying synaptic vesicle function at the pre-synaptic site of the corticostriatal synapse (PARKIN mutations. In particular, as compared with control neurons, the parkin-deficient neurons had a stronger response to the activation of D1-class dopamine receptors (These studies on the localization of parkin and its capacity to directly regulate excitatory transmission constitute an important step toward understanding the functions of parkin at the synapses . The com9 months . Decreasout mice . In the uishable . DA overout mice . Indeed,out mice . These rlevodopa . The autlevodopa ; nonethe synapse . Moreoveeceptors .This raises the possibility that parkin could be involved in D1 receptor hypersensitization after dopaminergic denervation, an important mechanism underlying the dyskinesia in iPD . AccordiTaken together, these findings suggest that parkin orchestrates a complex remodeling of corticostriatal synapses through multiple mechanisms. These mechanisms are likely to include changes due to DA depletion as well as those due to loss of parkin function at these synapses: alterations in the trafficking of glutamate vesicles at the glutamatergic pre-synapse and/or dysregulation of glutamate receptor levels on MSNs and/or dysregulation of D1-signaling. These mechanisms may be responsible for the precocious maladaptive plasticity of the corticostriatal synapses that leads to early dyskinesia .PARKIN mutation affects corticostriatal transmission, leading to early maladaptive changes in this glutamatergic synapse. This alteration could produce changes due to corticostriatal-induced depletion of DA at the synapses. What does this scenario imply for the development of specific and effective treatments? The only currently available drug for symptomatic treatment of dyskinesia is amantadine, an antiviral agent introduced in the early 1970s that also displays NMDA receptor antagonism (PARKIN patients and late-onset PD patients could be relevant for devising precision medicine strategies for parkin patients. The recent discovery that LID is caused by a distinct and stable group of striatal neurons (Since the parkin protein modulates glutamatergic receptor function and possibly glutamate release as well, it is conceivable that tagonism . With a tagonism . Moreove neurons  paves thPARKIN mutation.In conclusion, published data support the hypothesis of an early corticostriatal synaptopathy in parkin patients caused by parkin modulating the glutamatergic synapse. This hypothesis could provide a biological basis for studying the disease phenotype and lay the groundwork for the pharmacological treatment of PD patients with JS wrote first draft of the manuscript. FV and AC reviewed and critically 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.


Background: The current study examines the cross-sectional association between everyday discrimination and kidney function among older adults. Methods: We use cross-sectional data from a nationally representative sample of older adults to examine this relationship. Our measure of kidney function derives from the estimated glomerular filtration rate (eGFR) obtained by the Chronic Kidney Disease Epidemiology Collaboration equation, while our indicator of everyday discrimination is drawn from self-reports. Results: Results from our ordinary least squared regression models reveals that, after adjusting for demographic characteristics, everyday discrimination was associated with lower mean eGFR . The relationship between everyday discrimination and kidney function was not explained by cardiovascular, metabolic, or economic factors. Conclusions: Findings suggest this study suggest that everyday discrimination may be a unique risk factor for poorer kidney function among older adults. Because these findings are cross-sectional, additional research is needed to determine whether the observed associations persist over time.


Molecular Cancer, Wang and colleagues’ work revealed the common and histopathological subtype-specific circular RNA (circRNA) expression patterns between LUAD and LUSC and highlighted the important diagnostic potential of circRNAs in lung cancer [Few studies have used high-throughput RNA sequencing for functional characterization of patients with non-small cell lung cancer (NSCLC) including lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). In a study recently published in g cancer .p-value ≤0.05 and fold change ≥2), differential expression of 50 and 172 circRNAs was identified in tumor tissues of LUAD and LUSC, respectively. In addition, 26 circRNAs were commonly deregulated in both tumors. Moreover, another independent cohort of 67 lung cancer patients’ samples were subjected to experimentally validation of RNA-sequencing data. Results were consistent and validated the accuracy and robustness of the circRNA analysis pipeline.To obtain a comprehensive and accurate quantification of circRNAs, the authors analyzed high-throughput RNA sequencing data from 10 pairs of lung tumors and their adjacent normal tissues using five computational tools. Results revealed 17,952 circRNAs including 2593 previously unidentified circRNAs. Using stringent criteria  curve analysis of several selected circRNAs and revealed high diagnostic potential of hsa_circ_0077837 and hsa_circ_0001821 in discriminating NSCLC from normal tissues. Authors’ results also highlighted the potential application of hsa_circ_0001073 and hsa_circ_0001495 for diagnostic in the histopathological subtyping of NSCLC. Collectively, this study demonstrated that LUAD and LUSC as the major subtype of NSCLC share a common differential expression pattern, but also have their unique circRNA expression signatures. Identified circRNAs may serve as and new clinical tools for diagnosis of pathological subtypes of NSCLC and therapeutic intervention.


Dear Editor,The study by Terregino et al , outlineThe ECRS scale was tested with videos of students participating in objective structured clinical examinations (OSCEs) . CompariThe ECRS could also be used for a wide range of scenarios, including those appearing acutely medical. For example, a case about anaphylaxis allows the student to communicate with the patient and their family whilst prioritising acute management of the life-threatening condition. Additional notes could be added to the ECRS about how students balance emergency medical management with their communication towards the patient and family. This would allow students to act like physicians who manage medical problems alongside patient ideas, concerns and expectations. Focusing training on empathy ensures students display empathy in real-life scenarios as future doctors .Additionally, a modified ECRS could be used to practice other realistic clinical encounters including consultations across a language barrier. Although interpreters should be available for consultations, issues including administrative errors mean this may not happen. Non-native speakers may not pick up on empathetic phrases and therefore rely on nonverbal, compassionate behaviours that cross-cultural boundaries .To conclude, the ECRS appears to be a useful tool to assess medical student empathy in OSCEs to ensure students develop patient-centred communication skills . Whilst


Circulating tumor cells (CTCs) slough off primary tumor tissues and are swept away by the circulatory system. These CTCs can remain in circulation or colonize new sites, forming metastatic clones in distant organs. Recently, CTC analyses have been successfully used as effective clinical tools to monitor tumor progression and prognosis. With advances in next-generation sequencing (NGS) and single-cell sequencing (SCS) technologies, scientists can obtain the complete genome of a CTC and compare it with corresponding primary and metastatic tumors. CTC sequencing has been successfully applied to monitor genomic variations in metastatic and recurrent tumors, infer tumor evolution during treatment, and examine gene expression as well as the mechanism of the epithelial-mesenchymal transition. However, compared with cancer biopsy sequencing and circulating tumor DNA sequencing, the sequencing of CTC genomes and transcriptomes is more complex and technically difficult. Challenges include enriching pure tumor cells from a background of white blood cells, isolating and collecting cells without damaging or losing DNA and RNA, obtaining unbiased and even whole-genome and transcriptome amplification material, and accurately analyzing CTC sequencing data. Here, we review and summarize recent studies using NGS on CTCs. We mainly focus on CTC genome and transcriptome sequencing and the biological and potential clinical applications of these methodologies. Finally, we discuss challenges and future perspectives of CTC sequencing. Circulating tumor cell (CTC) studies began in 1869 Ashworth , which hPrevious CTC studies have mainly focused on the development of CTC enrichment technology  technology in NCBI PUBMED and found 19 CTC publications  or to delete immune cells (CD45−) , genome or transcriptome amplification, and sequencing and analysis Fig. . SeveralAfter CTC enrichment from the blood, zero to several hundred CTCs may be retained in a thousand to ten thousand background cells, resulting in a low efficiency for sequencing and analysis of these cell pools  CTC variation detection rate in a mean depth of 1500X of 14 CTCs. These authors concluded that there exists a high inter- and intra-patient heterogeneity in CTC mutational status  is also frequently altered during cancer evolution. Ni et al. surveyed the CTC CNVs from a small-cell lung cancer (SCLC) patient during sequential chemotherapy treatment and observed that the evolution of CNV was consistent along the therapeutic stage, indicating that the reproducible CNV pattern was not affected by drug treatment (Ni et al. Previous studies have highlighted particular molecular pathways involved with cancer metastasis, such as TGF-β signaling (Akhurst and Derynck WNT2 gene mediated the metastasis-associated survival signal, consistent with observations of the upregulation of multiple Wnt genes in pancreatic patients (Yu et al. Yu et al. first utilized single-molecule RNA sequencing and a mouse pancreatic cancer model and found that the The EMT of adherent epithelial cells to a migratory mesenchymal state has been implicated in tumor metastasis in pre-clinical models. Yu et al. characterized the dynamic cell fates in breast cancer CTCs and found an association of mesenchymal CTCs with disease progression (Yu et al. As shown in this review, CTC sequencing can now be used as an efficient liquid biopsy tool to investigate the spectrum of somatic variation and gene expression changes in primary, metastatic, and recurrent cancer patient tumors non-invasively. The somatic alterations could either be used to understand the origin, ITH, and evolution of tumors or to monitor disease progression during cancer treatment. The most important clinical implication of CTC sequencing is for personalized medicine, or so-called precision medicine, according to the variation spectrum detected, which indicates the selection of target therapy based on the CTC variation spectrum. With the development of CTC capture and SCS technologies, we can expect the establishment of more comprehensive cancer origins and evolution models in the near future. Moreover, more novel biomarkers or potential drug targets for cancer metastasis or drug resistance prevention or treatment can be identified through CTC sequencing.However, as mentioned above, sequencing a CTC genome or transcriptome faces technical challenges. Obtaining enough cells for library preparation and sequencing is the first critical step in CTC sequencing. However, various conclusions can be drawn from different studies based on different isolation method and cancer types. Some cancer types tend to generate more CTCs than other cancer types (Allard et al. More importantly, bioinformatics analyses of CTC sequencing data require additional quality evaluation and assessment, particularly the biases introduced during sample and sequencing library preparation Fig. . Allele Despite the development of advanced microfluidics approaches, several novel sequencing technologies show promise for solving the technological hurdles to CTC sequencing. For example, scientists can now perform in situ DNA or RNA sequencing, even on samples fixed on slides (Lee et al.


Both affective and blood pressure (BP) reactivity are associated with long term risk of chronic disease and mortality. Thus, understanding age-related changes in negative affect and BP responses to everyday demands is vital for promoting healthy aging. However, few studies have examined both psychological and BP reactivity simultaneously, which would provide more comprehensive understanding of regulatory processes at play. For the present study, 232 adults aged 50-70 years were assessed at baseline and 6 years later with ambulatory BP monitoring and momentary electronic diaries. Reactivity coefficients were output from multilevel models and used to test changes in negative affective and ambulatory BP reactivity to task demand, longitudinally. Results indicate that both systolic and diastolic BP reactivity increase with age, while negative affect reactivity does not change with age. Results are discussed in the context of life course theories of role strain and role changes and socioemotional theories of aging.


Microbial consortia execute collaborative molecular processes with contributions from individual species, on such basis enabling optimized molecular function. Such collaboration and synergies benefit metabolic flux specifically in extreme environmental conditions as seen in acid mine drainage, with biofilms as relevant microenvironment. However, knowledge about community species composition is not sufficient for deducing presence and efficiency of composite molecular function. For this task molecular resolution of the consortium interactome is to be retrieved, with molecular biomarkers particularly suited for characterizing composite molecular processes involved in biofilm formation and maintenance. A microbial species set identified in 18 copper environmental sites provides a data matrix for deriving a cross-species molecular process model of biofilm formation composed of 191 protein coding genes contributed from 25 microbial species. Computing degree and stress centrality of biofilm molecular process nodes allows selection of network hubs and central connectors, with the top ranking molecular features proposed as biomarker candidates for characterizing biofilm homeostasis. Functional classes represented in the biomarker panel include quorum sensing, chemotaxis, motility and extracellular polysaccharide biosynthesis, complemented by chaperones. Abundance of biomarker candidates identified in experimental data sets monitoring different biofilm conditions provides evidence for the selected biomarkers as sensitive and specific molecular process proxies for capturing biofilm microenvironments. Topological criteria of process networks covering an aggregate function of interest support the selection of biomarker candidates independent of specific community species composition. Such panels promise efficient screening of environmental samples for presence of microbial community composite molecular function. Microbial communities display cooperation and leverage on synergies, all embedded on the level of entangled molecular processes across individual species. Occurrence of such traits is tightly controlled via given species genotypes and environmental conditions, and centers on cost and benefit . ExtremeCooperation displays as higher order function, i.e. molecular processes executed by the community being not reducible to any of its individual species alone. Higher order molecular functions may alter local environmental parameters, in turn imposing a feedback on cooperation mechanisms, individual species abundance and specific involvement. Examples for such community properties include occupation of sites lacking viable conditions for individual species, division of labor, and fostering significant increase in metabolic flux .Microbial communities hold profound relevance in industrial processes. One example is bioleaching, i.e. extraction of base metals from sulfide ores . A seconRational design and optimization of community function was recently demonstrated for copper bioleaching . HoweverExtrapolation from a species inventory to capabilities of embedded community molecular function is limited, as resolution of species interaction on a molecular function level is needed. Next to allowing optimization of given consortia properties such knowledge promises particular utility in molecular function prospecting. Such scenario focuses on identification of communities in environmental samples exhibiting specific aggregate molecular function of interest .Recalling that increased efficiency in bioleaching resembles a composite molecular process in the sense of a microbial community aggregate function, a relevant proxy for such molecular configuration is given as molecular biomarkers. Use of molecular biomarkers is further supported by recently identified evidence that function and underlying molecular processes of a community may be preserved while the species composition of a community exhibits a dynamic flux . FurtherBiomarker identification approaches include explorative methods, involving comprehensive functional gene arrays as well as various omics tools. For example, Roume et al. integrated omics profiling results on molecular interaction networks for characterizing the impact of environmental conditions on microbial communities relevant in biological waste water treatment . On the M composed of n individual microbial species mi, mj, … mn which together constitute the community. A higher order function results out of a composite molecular process pi, principally grounded on the set of protein coding genes gi, gj of all community species, hence the community genome G.Analysis of molecular context is of particular value when aiming at characterizing composite function. Microbial community representation has to focus on the species set G for deriving the specific set of cross-species processes pi, pj responsible for triggering phenotypic community characteristics. A biomarker in its definition serves as proxy for the status of a molecular process. Having identified candidate processes pi, pj responsible for a molecular function of interest in turn allows selecting corresponding candidate biomarkers bi, bj, where bi monitors the state of process pi, and bj of pj, respectively. Recalling the finding of community stability in terms of composite function but not in terms of species composition, such a biomarker panel may offer generalization in prospecting of environmental samples. Further, such biomarker panel approach realized as multiplexed assay may prove efficient for high throughput analysis of environmental samples.The composite molecular process sees involvement of molecular functionality (gene products) from different species, needing modeling of the microbial community on the level of in silico workflow for identifying biomarker candidate panels aimed at reflecting aggregate microbial community properties. Starting with a reference inventory of a functionally relevant subset of G further combined with interaction information allows deriving a molecular process graph. On the graph level, topological parameters aiming at resembling molecular process proxies as well as species coverage considerations are used for biomarker candidate selection. The resulting candidate biomarker panel is thus specific for the function in focus but to some degree agnostic regarding actual species composition. Such candidate panel promises generic applicability in bioprospecting regarding a composite molecular function of interest.On this basis we propose an While the workflow is applicable for studying various instances of microbial community aggregate function we in the following focus on community cooperation in forming a biofilm microenvironment in copper bioleaching/AMD. Biofilms can be instantiated by certain individual microbial species, however, in most environmental settings multiple species contribute molecular function to this microenvironment. In selected scenarios such environment provides grounds for emergent community phenotypes, including significant increase in bioleaching efficiency or initiation of AMD –15.The workflow illustrated in In a first step scientific literature is screened for microbial communities reported as relevant in a given focus (as copper bioleaching/AMD) for cataloguing constituent species genomes. In addition, biological community aggregate molecular functions reported as relevant are extracted. In a second step, catalogued genes are assigned to community molecular functions. Third, an all-against-all ortholog map is created for the gene set assigned to the biological function term in focus (e.g. biofilm formation), followed by consolidating protein-protein interaction information for the set of orthologs (step 4). With a gene set and respective interaction data at hand a molecular process model approximating the function term is retrieved (step 5), on this basis deriving topological parameters of the interaction network for guiding biomarker candidate selection (step 6). Finally, candidate biomarkers are forwarded to experimental evaluation (step 7).A list of natural acid mine drainage sites, major industrial bioleaching operations as well as suitable laboratory scale experiments, in the following referred to as ‘sites’, was compiled by mining the National Library of Medicine (NCBI) PubMed scientific literature repository in publication title and abstract for the terms ‘bioleaching’ and ‘acid mine drainage’. For all identified sites respective mineral endowment was extracted. For industrial mining sites additional mineral composition data were sourced from the Mindat database . AltogetNext, scientific literature covering the sites was screened for inventorying involved microbial species. 133 microbial species could be retrieved, of which 42 are specifically associated with one or more of the 18 copper sites.For each of the copper site-associated microbial species gene-centric annotation was retrieved from public databases as described previously . BrieflyEach copper site-associated species was evaluated for availability of annotation data for assigning protein coding genes to functional groups (COG) , and forA matrix of biological function terms and genes of relevance in copper bioleaching was obtained by manual data extraction from scientific publications reporting species given in Leptospirillum ssp. Group IV ‘UBA BS’ and T. arsenitoxydans) were sourced from the NCBI RefSeq database.A comprehensive all-against-all ortholog screening of the 25 species holding COG annotation was executed using the InParanoid pipeline (version 4.1) with default settings . As inpuProtein interaction information was retrieved from STRING  and subsequently aggregated across all 25 species using ortholog screening results to obtain an interactome on the ortholog level. This procedure allowed inclusion of protein coding genes of species as such not represented in the STRING database on the level of identified ortholog sequences from respective species being covered in STRING. The resulting interaction matrix was used to derive an ortholog network.Topological parameters of the ortholog graph were calculated using the NetworkAnalyzer application of Cytoscape version 3.2.1 , networkFor evaluation of candidate biomarkers selected on the ortholog graph level resembling the composite process of biofilm formation available omics profiles were used. A scientific literature review identified omics profiles in scope of bioleaching/AMD providing a resolution on the level of protein coding gene products. Data sets of specific interest needed to resemble meta-transcriptomics or -proteomics profiles covering microbial communities exhibiting biofilms in various environmental or laboratory conditions.Leptospirillum and Ferroplasma species. To assess biofilm functional traits quantitative proteomics comparison of natural versus laboratory settings was conducted, complemented by determining biomass to estimate in situ chemoautotrophic production. Productivity in the laboratory biofilm community was nearly five times lower compared to its natural equivalent, being reflected in differential abundance of a range of metabolic proteins. The authors concluded that the differences in both production as well as protein expression profiles reflect metabolic stress in the laboratory biofilm. After optimization of laboratory culture conditions , community proteomics profiles resembled the environmental setting particularly in respect to expression of stress response proteins and community growth rates.Two data sets appeared particularly suitable with respect to biofilm formation serving as surrogate of bioleaching/AMD. The first study conducted by Belnap et al. covers proteomics profiling . The autFor calculation of differences in expression in the proteomics data the respective log2-ratios of abundance when comparing the optimized laboratory culture with a natural biofilm sample were subtracted from the respective abundance values comparing a standard laboratory culture with a natural biofilm. Respective fold changes are therefore indicative for the level of biofilm formation and are used in biomarker candidate evaluation with respect to prospecting the capacity of forming a composite biofilm microenvironment.A. ferrooxidans as well as L. ferriphilum and L. ferrooxidans, respectively. Further community members worth noting include Acidimicrobium ferrooxidans as well as several Acidiphilum, Ferroplasma and Sulfobacillus species, respectively. The study compared transcriptional profiles of natural planktonic and biofilm-associated consortia to gain insights into physiological differences between free and the sessile community fractions. This approach allowed a precise assessment of biofilm physiology determined on a genomic array of L. ferrooxidans, identifying as main contributors to the biofilm situation genes coding for quorum sensing, motility and chemotaxis, as well as matrix component production and transport.A second data source originates from a transcriptomics profiling study on bacterial communities isolated from the Rio Tinto acid mine drainage system  published by Moreno-Paz et al. . The comIn biomarker evaluation the fold changes (log2-ratios) are used contrasting planktonic and biofilm status in focus of biofilm formation capacity.L. ferriphilum in continuous vs. bioleaching conditions, the latter being grown on 2% (wt/vol) chalcopyrite [A third study conducted by Christel et al. compared a culture of copyrite . The autThis study serves as negative control experiment for candidate biomarker evaluation, utilizing fold changes (log2-ratios) comparing cultures in absence and presence of chalcopyrite.Protein coding genes addressed in the three experimental studies were assigned to the ortholog graph via Basic Local Alignment Search Tool 2 (BLAST2) , selectiA. ferrooxidans (identified in 11 sites), L. ferrooxidans (9 sites), L. ferriphilum (7 sites) and A. cryptum, A. ferrivorans, A. thiooxidans and F. myxofaciens (4 sites each). Each site further reports a diverse and distinctive array of less frequent species.An important prerequisite for candidate biomarkers to be used in microbial community screening is applicability in absence of prior information about specific community species composition. Therefore a meta-analysis approach is applied starting with an inventory of microbial communities being involved in either bioleaching or acid mine drainage. Consortia retrieved from 70 environmental sites in focus of bioleaching/AMD provide 133 individual species. 18 sites report on copper sulfite ores with in total 42 individual microbial species. The majority (32) is comprised of Bacteria, complemented by Archaea. The four most prevalent species are Manual literature curation of molecular features involved in molecular processes relevant in acid mine drainage and/or bioleaching together with their molecular functional annotation identifies 14 molecular function categories holding 1,696 protein coding genes . The Clu20 of the 26 COG terms are addressed in the AMD/bioleaching context, with specific function terms seeing varying assignment to COG terms. For instance, chemotaxis is associated with 5 COG terms, on a gene count level mainly involving cell motility (N) and signal transduction (T). Other terms are considerably more complex, specifically the function term biofilm formation involving 19 COG terms. Main contributions come from cell wall/membrane/envelope biogenesis (M), cell motility (N), signal transduction (T), intracellular trafficking and secretion (U) as well as extracellular structures (W). Other complex functions with diverse COG term involvement include quorum sensing together with toxicant and copper resistance. Genes lacking COG annotation or being assigned to the unspecific COG terms R  and S (Function Unknown) are not further considered.Related function terms map to similar COG term combinations. For biofilm formation, COG terms include cell wall biogenesis, signal transduction and trafficking/secretion categories. Comparable patterns of COG category involvement are found for quorum sensing and chemotaxis, reflecting close biological relationship.For deriving a biomarker candidate panel capturing major molecular aspects of relevance in both AMD and copper bioleaching, function terms covering a broad set of COG terms appear preferential, ideally further seeing broad involvement of microbial species. Of the 14 function terms, biofilm formation exhibits particularly broad COG term assignment as well as species coverage across the original catalogue of 25 copper mineral associated microbes, providing all ingredients for establishment of higher order molecular function. Furthermore, from a biological function perspective, establishment of biofilms is relevant in both acid mine drainage as well as bioleaching. As demonstrated, enrichment of consortia with microbes particularly competent in formation of biofilms significantly boosts bioleaching process efficiency . BiofilmAccordingly, the set of genes assigned to the aggregate function of biofilm formation is used as basis for deriving a protein coding gene interaction network, subsequently serving for biomarker panel selection.According to data retrieval and functional category assignment 272 protein coding genes are identified in the molecular context of biofilm formation. Ortholog screening of this set in the genomes of the 25 species provides 3,106 orthologs, identifying at least one ortholog for all but 23 genes. 57 of the 272 genes hold orthology to other members of the biofilm gene set and are removed to avoid biases from multiple sequence representation in subsequent molecular network retrieval.Ortholog information allows utilizing interaction information across species to derive an ortholog graph, thereby complementing interaction data by aggregation. This procedure enables inclusion of interaction information for finally 191 unique molecular features of the biofilm gene set  in a sinL. ferrodiazotrophum, a diazotrophic mesophilic bacterium also found in AMD biofilms [L. ferrodiazotrophum efficiently fixes nitrogen, but its genome lacks annotation including COG term assignment and interaction data in STRING. The nitrogenase cassette is highly homolog to features found in other diazotrophic bacteria such as L. ferrooxidans and Acidithiobacillus ssp., i.e. the cassette is considered at the ortholog level.The ortholog network allows capturing molecular function of species as such not included in a species set. Representative for such an organism is biofilms . L. ferrThe ortholog graph serves as basis for identifying biomarker candidates according to network topology. Topological parameters characterizing the centrality of a given node capture properties related to the functional importance within its network context. Specifically, node degree and stress centrality resemble properties prone to biomarker candidate selection. Both centrality measures reflect affectedness of state changes in molecular processes, hence serving as proxies for molecular process status. Nodes with high degree centrality establish a large number of interactions, i.e. serve as network hubs . Nodes eThe relevance of degree and stress becomes apparent from Accordingly, biomarker candidates from the ortholog graph of biofilm formation showing high degree and stress centrality are deemed to serve as relevant proxy for the entire process, and promise sensitivity in monitoring the presence of biofilm formation.Relevance of network topology aspects is evaluated using proteomics profiles published by Belnap et al., contrasting a generic and an optimized laboratory setting with environmental samples in focus of biofilm formation efficiency . ProteomAnnotation of nodes of the ortholog graph with fold change values assigned to biofilm formation in combination with computation of the topological parameters degree as well as stress centrality allow determining significance in difference of graph measure distributions. Degree as well as stress centrality are found to be different for nodes holding a significant fold change compared to nodes not found to be affected according to the proteomics data  positive correlation is identified , aside lBased on the observed association of degree and stress centrality with a probability of abundance change under varying biofilm conditions, a rank score of both graph measures and species coverage is determined. Each individual parameter is ranked starting with the maximum value, with the rank score reflecting the mean of the sum of each individual parameter rank for a given protein. L. ferrooxidans, significant orthology of sequences selected in the biomarker panel render fold changes as indicative for biomarker expression on the background of the ortholog network.For evaluation of the biomarker candidate panel two omics studies appear particularly useful due to providing i) substantial coverage of the entire molecular feature space represented in the biofilm network, and ii) showing differential readout for biofilm formation under varying conditions. The first study conducted by Moreno-Paz et al. resembles a prospecting situation and reports transcriptomics profiling comparing planktonic versus biofilm associated bacterial consortia in a natural AMD site . The stuA monitoring/process optimization situation can be approximated via comparing proteomics data of the generic and the optimized laboratory condition as provided in Belnap et al. . With opL. ferrooxidans cultivated in an agitated-tank bioreactor [The third data set prepared by Christel et al. compared gene expression of a continuous and a bioleaching  culture of oreactor . This sporeactor  and Moreoreactor ), and isA. ferrooxidans on pyrite [A further data set provided by Vera et al. implemented shotgun proteomics for characterizing early biofilm formation of n pyrite . In thisThe most prominent function category involved in the biomarker set is chaperones with four protein-coding genes. High ranking in degree centrality reflects broad involvement in folding and arrangement of macromolecular structures as well as their propensity to associate with hub proteins. This property meets with the processes of biofilm formation, but questions specificity, as activity of these proteins is supposedly relevant in a variety of molecular processes beyond the aggregate function in focus.Still, given chaperones hold specific evidence in the biofilm context in an acidic environment. GroEL chaperones are generally known as stress response proteins and are frequently found in acidophilic bacteria  when grown on elementary sulfur. A more specific role of the protein family is modulation of fatty acid components of bacterial cell walls deemed essential during biofilm maturation . The putA second prominent function term covers central biofilm components involved in the synthesis of extracellular polymeric substances. Members include a uridine diphosphate (UDP)-glucose 4-epimerase converting uridine diphosphate galactose to UDP-glucose, a precursor for the osmoprotectant trehalose, as well as several exopolysaccharides , 39. FurThe third term set covers chemotaxis, sensing and motility, of apparent relevance in structuring the community microenvironment in a biofilm. For dimethyladenosine transferase a more specific function in quorum sensing was discovered and involves environmental adaptation through cross-species synchronization . As secoTaken together, the panel members represent core functional elements required in establishing a biofilm microenvironment, covering chemotaxis, motility, quorum sensing as well as the production of extracellular matrix components. The prominent representation of chaperones of the GroEL-Dnak-GrpE system not only points to adaptation processes allowing communities to establish in the extreme environments, but also reflects the need for a proper orchestration of the biofilm formation processes . Genes fStill, the exposed network topology of the panel members being hubs and connectors and on top being encoded in a wide range of species, in various instances covering more than 20 of the in total 25 microbial species involved, may in part be driven by annotation biases of network nodes and interactions, at present not allowing for evidence-based correction. Aside this aspect the need for specialist species for driving aggregate molecular processes is to be addressed. Analyzing species coverage of nodes in the network neighborhood of degree and stress centrality hubs sees more sparse species coverage, indicating the need for marginal consortia members on the molecular process level . As examMolecular features involved in biofilm formation are identified in a range of organisms commonly isolated in AMD and bioleaching sites. For a subset of organisms a sufficient depth of annotation is available both on functional as well as on protein interaction level. Lack of annotation per design removes a subset of protein coding genes from integrative analysis, although eventually resembling excellent biomarker candidates. However, coding regions displaying no orthology to annotated sequences of an extended species set retrieved in a meta-analysis approach may be fairly specific for certain species. In consequence, biomarkers from such subset may lack generalization capacity, in turn limiting sensitivity in a species-agnostic prospecting approach.Available interaction data allow deriving an ortholog network to approximate the community molecular process of biofilm formation. By integrating orthology information in the network some major limitations, given by unknown or incompletely characterized genes, lack of functional annotation as well as of interaction data can be addressed to some extent. The interaction data source used in this work provides interactions holding experimental evidence and candidate interactions resting on computational inference. With lack of completeness of experimentally derived interactomes together with biases introduced by experimental protocols, inference of interactions from heterogeneous data sources in part addresses given limitations and biases. This approach at the same time introduces probabilistic assignments. In consequence, resulting networks primarily serve hypothesis generation, demanding subsequent experimental evaluation. On the other hand, correlation of abundance of sequences with unknown function and annotated nodes of a mechanistic network model may support candidate function assignment .Combining various functionalities deemed necessary for effective biofilm formation in a multimarker panel promises improved sensitivity as well as specificity. The functional diversity of the process of forming a biofilm microenvironment, specifically beyond enzymes, points to an important aspect for selecting a comprehensive network model. For example, approaches restricted to metabolic networks by e.g. utilizing KEGG orthology (KO) terms of enzymes (nodes) connected by metabolites (edges) cover less than half of the network nodes included in the biofilm orthology graph . In addiThe workflow discusses candidate biomarker retrieval for prospecting of an aggregate biological function executed by microbial consortia in environmental samples. The approach leverages on cross-species interaction consolidated as ortholog network, and utilizes topological properties of network nodes for molecular feature selection.Identification of candidate panel members is based on a molecular process model tailored at representing the molecular context of biofilm formation. The candidate biomarkers are selected according to a rank score of degree and stress centrality as well as species coverage, thus focusing on prevalent proteins being deemed essential for establishing a biofilm microenvironment. Evaluating candidate biomarkers via utilizing public domain omics profiles indicates relevance in biofilm formation. Applicability of the method is not restricted to biofilm prospecting in copper mineral decomposition, but is expandable to alternative microbial community functions including as examples biofertilization in agriculture or microbial communities serving in oil spill bioremediation.de novo functional characterization of unknown species.Practical implementation on the mRNA level, e.g. via quantitative polymerase chain reaction (qPCR)-based methods, demands conserved sequence regions as probes. The approach of defining a set of biomarkers designed to screen for a given molecular function being at least to some extent independent of the executing microbes bears the promise to significantly facilitate prospecting of novel bacterial consortia. The precise composition of consortia is in most cases unknown at the point of screening. A species-robust screening tool supersedes the need for any prior knowledge on likely species occurrence. In line with prospecting goals a biomarker panel allows a first evaluation of samples, and in case of positive biomarker readout providing grounds towards a more detailed molecular analysis e.g. via metagenomics. This is particularly useful in cases where potential prospecting areas are large and diverse, or where a fast and frequent read-out, e.g. in a monitoring situation, is needed. Further, the approach facilitates the targeted discovery of new species in a functional context in focus, particularly if combined with novel techniques such as methods for In conclusion, interaction networks offer a methodology for characterization of community aggregate molecular functions via integrating individual species contributions into a molecular process network. Next to adding to our understanding of molecular mechanisms of aggregate function such networks allow selecting candidate biomarkers grounded on network topology characteristics. Such biomarkers promise support in microbial function prospecting as well as in industrial process monitoring in a range of application areas, with bioleaching and bioremediation as examples.S1 TableSpreadsheet “sites”: Copper site-specific scientific references; spreadsheet “species”: Microbial species-specific scientific references; spreadsheet “functions”: key scientific references covering molecular function terms.(XLSX)Click here for additional data file.S2 TableProvided is the protein name together with identifiers from database references of UniProt, PATRIC and IMG.(XLSX)Click here for additional data file.S3 TableThe listing holds genes assigned to the functional term biofilm formation embedded in the ortholog network, respective topology measures of degree and stress centrality, species coverage, and ranked sum of topological parameters and species coverage.(XLSX)Click here for additional data file.


In Sub-Saharan Africa, there is an increase in trypanosome non-susceptibility to multiple trypanocides, but limited information on judicious trypanocide use is accessible to smallholder farmers and agricultural stakeholders in disease endemic regions, resulting in widespread multi-drug resistance. Huge economic expenses and the laborious nature of extensive field studies have hindered collection of the requisite large-scale prospective datasets required to inform disease management. We examined the efficacy of community-led data collection strategies using smartphones by smallholder farmers to acquire robust datasets from the trypanosomiasis endemic Shimba hills region in Kenya. We used Open Data Kit, an open-source smartphone application development software, to create a data collection App.Our study provides proof of concept for the viability of using smartphone Apps to remotely collect reliable large-scale information from smallholder farmers and veterinary health care givers in resource poor settings. We show that these datasets can be reliably collated remotely, analysed, and the findings can inform policies that improve farming practices and economic wellbeing while restricting widespread multi-drug resistance. Moreover, this strategy can be used to monitor and manage other infectious diseases in other rural, resource poor settings.The online version of this article (10.1186/s13104-019-4198-z) contains supplementary material, which is available to authorized users. Trypanosoma congolense, Trypanosoma brucei, and Trypanosoma vivax) is endemic to Shimba hills, in Kwale County, Kenya, a resource poor setting inhabited by smallholder farmers [2 (23% of the country), putting livestock and people at risk of infection [Animal African trypanosomiasis (AAT), a life-threatening neglected tropical disease that affects cattle, goats and sheep, caused by trypanosomes parasites  , 22. IntAlthough performing financial transactions using mobile phones is predominant , there iCommunity engagement when implementing mobile phone-based strategies is important for gaining trust, educating on use and importance of data collected, and easy reporting of feedback . Kwale CThis pilot study was part of a larger study evaluating evolutionary markers of trypanocide non-susceptibility conducted among two smallholder livestock farming communities of different ethnicities from Mbegani and Kizibe in Shimba hills, Kwale County, Kenya . We obtaWe used Open Data Kit (ODK) an open-source smartphone App development software to create a data collection App . ODK hasWe used multiple correspondence analysis (MCA), a data reduction technique which permits the identification of complex patterns in a dataset of categorical variables , to deteWe conducted a pilot study on the efficacy of a community-led data collection approach by conveniently sampling from two communities of smallholder farmers (n = 47) from: Kizibe (n = 23) and Mbegani (n = 24) to acquire information on trypanocide non-susceptibility and perceptions of smallholder farmers on trypanosomiasis. There was a disproportionately lower number of female smallholder farmers , who also had less cattle ownership  compared to their male counterparts Fig. a. These A large proportion of smallholder farmers and veterinary health care givers interviewed  welcomed the smartphone App developed on the ODK platform as a method of trypanosomiasis data collection. The farmers with dissenting views about using the App were illiterate. Our multiple correspondence analysis results show variation between the two communities from the overall mean, which would not be detected if knowledge on community ethnicity was not considered.Kwale is one of the most resource poor Counties in Kenya. Kizibe and Mbegani, in Shimba hills, Kwale, are trypanosomiasis endemic regions with varying economic capacities and different farming practices, which have made understanding temporal trends in trypanocide non-susceptibility challenging. Previous studies have used generalised cross-sectional survey approaches that lumped together findings from different communities in Kwale making it challenging to pinpoint subtle differences in farmers perceptions and disease management practices . We estaWe used multiple correspondence analysis (MCA) to highlight subtle variation in farming practices by the smallholder farmers in Kizibe and Mbegani, which could be used to inform trypanosomiasis management. Similar approaches have been used to assess animal African trypanosomiasis (AAT) vulnerability in cattle-owning communities of sub-Saharan Africa . Our resMobile phone affordability and technology uptake has increased tremendously in Kenya , suggestThe limitation in this pilot study was focus on a restricted set of questions to establish proof of concept, but other pertinent research questions based on study design can be included for prospective research. We also anticipate some recall bias during data collection. We currently underpowered to estimate the extent of this bias, but anticipate that a much larger dataset from prospective data collection will reduce confounding during analysis. This study was conducted using datasets from smallholder farmers due to insufficient data from the small number of veterinary healthcare workers. Another setback was the intermittent mobile network coverage by some mobile service providers. We were able to circumvent this challenge by transmitting information to the central database once connected to a strong network.Additional file 1. Questionnaire used to develop the ODK-based mobile App. The App questions were divided into two: (i) questions to animal health care workers (ii) questions to smallholder farmers.


The addictive brain undergoes dramatic changes as a result of drug exposure. A growing body of evidence suggests a wide range of brain areas, including ventromedial prefrontal cortex (VMPFC), orbitofrontal cortex (OFC), dorsolateral prefrontal cortex (DLPFC) and other subcortical regions, namely anterior cingulate cortex (ACC), striatum, and anterior insula, are involved in drug-related cue processing  over left VMPFC could change the cue-related FC, which was examined by fMRI scans before and after cTBS (Kearney-Ramos et al., These understandings were addressed in a recent study published on the journal of Another extensive line of research has demonstrated VMPFC-limbic connections are altered in addicts (Ma et al., In conclusion, the current study demonstrated that cTBS of VMPFC-limbic circuits could change drug cue reactivity. Their results are likely to inspire more therapeutic interventions targeting drug-bias attention and self-awareness to modify previously inappropriate rewarded behavior for this chronic repeated relapsing disease. The approach combining fMRI with rTMS will be widely used to understand the underlying pathophysiologic neural circuits, and to design individualized treatment intervention. Nevertheless, although such evidence seems to be informative to us, it cannot effectively reduce drug-induced craving or relieve addiction syndrome. Our previous results indicated more participants respond to five repeated sessions with a better effect on craving in comparison to those who only receive a single session (Shen et al., All authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication.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.


Middle-age and older adults vary widely in their physical health. This symposium describes studies that identify diverse daily experiences that account variation in health using multiple indicators of well-being . Fingerman et al. assessed daily TV viewing among older adults and found that more frequent television watching was associated with poorer physical health, worse health behaviors, and less energy expenditure . Leger et al., examined links between daily affect and sleep. Greater variability in daily positive affect is associated with fewer hours of sleep and greater morning tiredness even after adjusting for mean levels of affect. Luong et al. examined links between daily stress and affect. Interpersonal stressors were associated with greater affect reactivity than non-interpersonal stressors and links were reduced among older adults. Birditt et al. assessed links between daily social interactions and cardiovascular reactivity. More frequent social interactions and negative social interactions were associated with increased heart rate and links varied by gender and race. Polenick et al. examined links between daily social interactions and salivary DHEA-S (a marker of the stress response). Positive interactions predicted greater DHEA-S over the course of the day and links between negative interactions and DHEA-S varied by age group such that younger individuals appeared to be more reactive. These studies offer important clues regarding how daily experiences get under the skin to influence health and well-being.


OBJECTIVES/SPECIFIC AIMS: Listening effort is needed to understand speech that is degraded by hearing loss and/or a noisy environment. Effortful listening reduces cognitive spare capacity (CSC). Predictive contexts aid speech perception accuracy, but it is not known whether the use of context reduces or preserves CSC. Here, we compare the impact of predictive context and cognitive load on behavioral indices of CSC in elderly, hearing-impaired adults. METHODS/STUDY POPULATION: Elderly, hearing-impaired adults listened in a noisy background to spoken sentences in which sentence-final words were either predictable or not predictable based on the sentence context. Cognitive load was manipulated by asking participants to remember either short or long sequences of visually presented digits. Participants were divided into low or high cognitive capacity groups based on a pretest of working memory. Accuracy and response times were examined for report of both sentence-final words and digit sequences. RESULTS/ANTICIPATED RESULTS: Preliminary results indicate that accuracy and response times for both words and digits were facilitated by sentence predictability, suggesting that the use of predictive sentence context preserves CSC. Response times for both words and digits and accuracy for digits were impaired under cognitive load. Trends were similar across high and low cognitive capacity groups. The preliminary results support the idea that habilitation strategies involving context use could potentially support CSC in elderly, hearing-impaired adults. DISCUSSION/SIGNIFICANCE OF IMPACT: These preliminary results support the concept that habilitation strategies involving context use could potentially support CSC in elderly, hearing-impaired adults.


While caloric restriction (CR) provides highly robust improvements to longevity and health, dietary restriction of the essential amino acid methionine can provide similar benefits including improved metabolic function and increased longevity. Despite these similarities between CR and methionine restriction (MR), there is growing evidence to suggest they may be mediated by different mechanisms that require further elucidation. The sulfur side-chain of methionine is highly prone to oxidation, even in vivo, with redox changes of these residues potentially altering protein function and interfering with its use as a substrate. An entire family of enzymes, methionine sulfoxide reductases, have evolved in aerobic organisms to regulate the redox status of methionine. We tested the role of methionine sulfoxide reductase A (MsrA) in the physiological and metabolic benefits of MR. After three months of MR, mice lacking MsrA (MsrA KO) showed significant loss of weight, including both fat and lean mass, in comparison to wild-type mice under MR. Both MsrA KO and wild-type mice responded to MR with improvements to both glucose and insulin tolerance. However, MR MsrA KO mice showed lower HbA1c and reduced leptin compared to MR wild-type mice. Overall, our results show mice lacking MsrA have a stronger response to MR suggesting that methionine redox may play an important role in some of the mechanisms responsible for these metabolic outcomes. Further studies clarify whether MsrA could also be a potential regulator of the longevity benefits of MR.


In meiosis, DNA break formation and repair are essential for the formation of crossovers between homologous chromosomes. Without crossover formation, faithful meiotic chromosome segregation and sexual reproduction cannot occur. Crossover formation is initiated by the programmed, meiosis‐specific introduction of numerous DNA double‐strand breaks, after which specific repair pathways promote recombination between homologous chromosomes. Despite its crucial nature, meiotic recombination is fraud with danger: When positioned or repaired inappropriately, DNA breaks can have catastrophic consequences on genome stability of the resulting gametes. As such, DNA break formation and repair needs to be carefully controlled. Within centromeres and surrounding regions , meiotic crossover recombination is repressed in organisms ranging from yeast to humans, and a failure to do so is implicated in chromosome missegregation and developmental aneuploidy. (Peri)centromere sequence identity and organization diverge considerably across eukaryotes, yet suppression of meiotic DNA break formation and repair appear universal. Here, we discuss emerging work that has used budding and fission yeast systems to study the mechanisms underlying pericentromeric suppression of DNA break formation and repair. We particularly highlight a role for the kinetochore, a universally conserved, centromere‐associated structure essential for chromosome segregation, in suppressing (peri)centromeric DNA break formation and repair. We discuss the current understanding of kinetochore‐associated and chromosomal factors involved in this regulation and suggest future avenues of research. Fusion of two gametes upon fertilization reconstitutes the diploid genome of the zygote. Meiosis relies on the same basic molecular machinery as mitosis but requires additional dedicated events ‐sized regions directly adjacent to centromeres  pericentromeres prevented? Ectopic targeting experiments by Robine et al.  revealedTaken together, a localized regulation of Red1 function within budding yeast pericentromeres could play a role in controlling DSB formation, in a manner that might share foundational principles within the fission yeast system. However, clear differences between fission and budding yeast regulation of DSB formation and repair exist. Most importantly, whereas in fission yeast, DSB regulation involves cohesin (Nambiar & Smith, 4As discussed here, fascinating new aspects of localized control of meiotic DSB formation and repair have recently been revealed. Despite structural differences within pericentromeres, a common thread is a key role for cohesin in controlling local DSB formation and repair (Nambiar & Smith,