To establish whether prevalence and severity of long-COVID symptoms vary by industry and occupation.We used Office for National Statistics COVID-19 Infection Survey (CIS) data (February 2021–April 2022) of working-age participants (16–65 years). Exposures were industry, occupation and major Standard Occupational Classification (SOC) group. Outcomes were self-reported: (1) long-COVID symptoms and (2) reduced function due to long-COVID. Binary (outcome 1) and ordered (outcome 2) logistic regression were used to estimate odds ratios (OR)and prevalence .Public facing industries, including teaching and education, social care, healthcare, civil service, retail and transport industries and occupations, had the highest likelihood of long-COVID. By major SOC group, those in caring, leisure and other services  had substantially elevated odds than average. For almost all exposures, the pattern of ORs for long-COVID symptoms followed SARS-CoV-2 infections, except for professional occupations  . The probability of reporting long-COVID for industry ranged from 7.7%  to 11.6% (teaching and education); whereas the prevalence of reduced function by ‘a lot’ ranged from 17.1%  to 22%–23% (teaching and education and armed forces) and to 27% (not working).The risk and prevalence of long-COVID differs across industries and occupations. Generally, it appears that likelihood of developing long-COVID symptoms follows likelihood of SARS-CoV-2 infection, except for professional occupations. These findings highlight sectors and occupations where further research is needed to understand the occupational factors resulting in long-COVID. SARS-CoV-2 infection and COVID-19 mortality in the UK varied by occupational group; yet it is not known if any occupational groups are more susceptible to long-COVID than others.This is the first study to examine how prevalence of long-COVID and its impacts on functional capacity differ by industry and occupational groups.Prevalence of self-reported long-COVID increased with time across all exposure groups and mostly followed SARS-CoV-2 infection trends; with the exception of professional occupations  that demonstrated notable differences in the direction of odds of long-covid when compared with odds of SARS-CoV-2 infection.Those working in teaching and education, and social care industries showed the highest likelihood of having long-COVID symptoms. The exact same pattern was observed when analysis was performed using occupational groups. When we used Standard Occupational Classification groups, the likelihood was higher in Caring, leisure and other services.The findings contribute to the evidence base that long-COVID differences occur across industries and occupations, provides insights for employees, employers, occupational and healthcare for the industries and occupations that may need additional support for return-to-work policies and highlights sectors and occupations where further research is needed to understand the mechanisms resulting in long-COVID, how occupational factors influence the risk of developing long-COVID or interact with long-COVID to increase the impact on activities and where further protective measures are needed.In the UK, the risk of SARS-CoV-2 infection and COVID-19 mortality has varied by occupational group, although differences appear to have declined over the duration of the pandemic.et al showed that almost half of the study respondents suffering with long-COVID required a reduced work schedule compared with preillness.Being out of work is associated with poor healthLong-COVID has a disproportionate impact on groups already disadvantaged in terms of work and health. Returning to work is part of rehabilitation from illness and is important to those recovering from long-COVID.Our study aims to establish whether the prevalence and severity of long-COVID varied by occupation and if any differences are in excess of differences in the risk of SARS-CoV-2 infection.We used data from the Office for National Statistics (ONS) COVID-19 Infection Survey (CIS) from February 2021 (long-Covid questions first included in CIS) to the end of April 2022. The CIS began in April 2020, used random sampling and aimed to be representative of the UK population. Repeated PCR testing for all participants was carried out during weekly home visits for the first month of entry to the survey, and then monthly until 30 April 2022 when testing samples were sent and returned by post and survey questions answered on-line or by phone.10.1136/oemed-2023-108930.supp1Supplementary dataAnalyses were restricted to participants aged ≥16 to ≤65 years old on the day of their first home visit and to those who turned 16 during the study period and had answered long-COVID questions at least once.We used the first available observation per occupational exposure and other covariates, assuming no change across time. In sensitivity analyses, we estimated panel models to allow for time variation.We used three different industry and occupational groupings. Industry was classified based on the ONS sector groupings used within CIS.Primary outcome measures were self-reported (1) cases of long-COVID and (2) reduced function  due to long-COVID among those reporting long-COVID, using the following CIS questions:Self-reported long-COVID: ‘Would you describe yourself as having ‘long-COVID’, that is, you are still experiencing symptoms more than 4 weeks after you first had COVID-19, that are not explained by something else?’. For our primary analysis, we reported the n (%) of people who self-reported having long-COVID in at least one survey, irrespective of any COVID-19 test result or previous infection reported.Self-reported reduced function (severity) of long-COVID: ‘How much did long-COVID reduce your ability to carry out daily activities?’ Respondents could answer 0: not at all, 1: yes, a little, 2: yes, a lot. This analytical sample was restricted to those self-reporting long-covid symptoms.Analysis was completed for the two outcome variables using all three exposure groups, irrespective of whether they have tested or reported being previously infected by SARS-CoV-2. For comparison, we also performed analyses using the outcome ‘SARS-CoV-2 infection (self-reported or positive PCR test).Sample characteristics were summarised using frequencies and proportions. Descriptive statistics were used to show the change in prevalence of self-reported long-COVID over time. Binary (outcome 1: long-COVID symptoms) and ordered (outcome 2: reduced function due to long-COVID) logistic regression models with robust standard errors to account for heteroskedasticity were used. For the binary logistic regression analyses, data were collapsed by person using the first reported values for exposure and covariates and taking ‘at least one reported episode of long-COVID’ as the outcome. For the ordered logistic regression, the highest reported value for the outcome variable  was used per person. We used effects coding where ORs are contrasted to each group mean compared with the grand mean showing the likelihood of having long-COVID in each group. To assess for potential confounding or mediating differences, we estimated unadjusted and fully adjusted models for all the covariates .Weighting was not used as the CIS derived weights are cross-sectional. Testing for patterns of missingness in the outcome variable did not show any differences . ParticiSensitivity analyses were conducted using two additional analytical samples: (1) those with a positive PCR test and (2) those with a positive PCR test and/or self-reported infection before entering the CIS. Furthermore, using the specifications of our main sample, a panel dataset was created where multiple participant visits and records were merged to create a dataset with one observation per month per person to allow for panel analysis that accounts for variation across time for time-varying variables, for example, ccupation, employment status . We usedAll analyses were calculated by using STATA V.17.Our final analytical samples included 323 766 individuals by industry, 277 487 individuals by occupational group and 307 363 individuals by major SOC group . Online Monthly prevalence of self-reported long-COVID and SARS-CoV-2 infection across time and across all three occupational exposures, demonstrates an increase of the prevalence of self-reported long-COVID with time, following similar trends in SARS-CoV-2 infection . PrevaleIn the unadjusted model , teachinBy occupational group, in the unadjusted model most demonstrated higher likelihood of having long-COVID; however, for some CIs are wide . SimilarUsing major SOC group as the exposure, in the unadjusted model , ORs forComparing the likelihood of long-COVID with the likelihood of SARS-CoV-2 infection  demonstrIn order to examine the extent to which long-COVID symptoms impacted daily activities , we used a reduced sample for all three exposure groups with analyses restricted to those self-reporting at least one long-COVID symptom over the study period. Final analytical samples were 30 543 participants for industry, 25 888 participants for occupational groups and 28 979 participants for major SOC groups .In the unadjusted model  all induAnalysis by occupational groups , similarBy major SOC group, in the unadjusted model  for almoPrevalence of reduced function by a ‘a little’ ranged between 47% and 49% for all three exposure groups . PrevaleAnalysis run on the specified two additional samples, that is, (1) confirmed PCR test or (2) confirmed PCR test or prior report of infection  did not For long-COVID, ORs from the panel analysis were relatively larger compared with our main analysis for almost all industrial and occupational groups. The opposite was observed for major SOC groups, although the magnitude of the ORs were closer to those in our main sample. Similar patterns were observed for the outcome of reduced daily function , suggestThe prevalence of self-reported long-COVID increased through the study period. The social care and education industry (~6%) and hospitality (~4%) occupations recorded the highest prevalence. Industries and occupations with high levels of office-based working , displayed lowest odds of long-COVID compared with more public facing industries, including teaching and education, social care, healthcare, civil service, retail and transport industries and occupations. By major SOC group, those in caring, leisure and other services  and education displayed  substantially higher odds than average. The likelihood of reporting long-COVID symptoms followed the trend of risk of infection for most industries, occupational groups and SOC groups. The exception was for professional occupations, where the odds of reporting long-COVID symptoms was higher than that of infections. Indications of differing patterns between infection and long-COVID were also observed in other occupational groups, but uncertainty was high. Long-COVID symptoms impacted almost half of all affected participants’ ability to do daily activities by ‘a little’. Participants in healthcare and social care industries were most impacted by ‘a lot’. Similarly, occupational groups most affected were public-facing occupations with those working in transport occupations having the highest prevalence of all (30%), followed by the not working (28.1%) and personal care (27.4%). For SOC groups the highest prevalence (24%) was observed in caring, leisure and other services and process plant and machine operatives.To our knowledge, no other studies examine long-COVID across industries and/or occupations for direct comparisons. However, previous work estimated that ~10% of people infected with SARS-CoV-2 may have significant postacute or chronic symptoms persisting >12 weeksOur study has several strengths. We used a large nationally representative dataset to examine prevalence and risk of long-COVID across industries, and occupations, which fills an important knowledge gap. Our analysis included a relatively long study period (February 2021–April 2022), allowing us to examine trends over time. Some limitations should be noted. Any analysis by occupation/industry requires aggregation of different types of workers. It is possible that analyses mask differences within particular sectors or occupations. Opting to use effect coding allows to compare each exposure group to the grand mean. Other methodological approaches may likely produce slightly different results driven by multiple comparison adjustment testing. Furthermore, differences in mitigation and protective measures across countries may lead to different results and comparable nationally studies are needed. In our main analyses, we used occupational status at the beginning of the study period, which may mask effects due to changes in employment status. However, our sensitivity analysis using panel data addressed this. At the time of data collection, there was no universal definition of long-COVID, and the results rely on self-reported measures which are prone to bias. It is possible that bias in this measure is related to occupation or due to other health condition. Furthermore, the discrepancy in duration of symptoms asked in the CIS  vs the latest WHO definition  means that some overestimation of cases may have occurred.The SARS-CoV-2 pandemic created new challenges for workers, employers, occupational and public health. Long-COVID is compounding to these challenges, mainly related to sustaining employment and return-to-work for those affected. Our findings show that long-COVID differences are evident across industries and occupations. Professional occupations, including jobs in teaching and education, IT, welfare and healthcare, demonstrated that the risk of infection may not be the only driving force. Our findings of increased risk of long-COVID in the health and social care industries support that some health conditions are prescribed as industrial diseases


Ultraviolet (UV) radiation directly affects plants and microorganisms, but also alters the species-specific interactions between them. The distinct bands of UV radiation, UV-A, UV-B, and UV-C have different effects on plants and their associated microorganisms. While UV-A and UV-B mainly affect morphogenesis and phototropism, UV-B and UV-C strongly trigger secondary metabolite production. Short wave (<350 nm) UV radiation negatively affects plant pathogens in direct and indirect ways. Direct effects can be ascribed to DNA damage, protein polymerization, enzyme inactivation and increased cell membrane permeability. UV-C is the most energetic radiation and is thus more effective at lower doses to kill microorganisms, but by consequence also often causes plant damage. Indirect effects can be ascribed to UV-B specific pathways such as the UVR8-dependent upregulated defense responses in plants, UV-B and UV-C upregulated ROS accumulation, and secondary metabolite production such as phenolic compounds. In this review, we summarize the physiological and molecular effects of UV radiation on plants, microorganisms and their interactions. Considerations for the use of UV radiation to control microorganisms, pathogenic as well as non-pathogenic, are listed. Effects can be indirect by increasing specialized metabolites with plant pre-treatment, or by directly affecting microorganisms. Life on Earth is exposed to the light spectrum ranging from ultraviolet-B (UV-B) to infrared wavelengths (295–2500 nm), hereafter referred to as natural radiation. The ultraviolet (UV) part of the electromagnetic spectrum comprises three classes: UV-C (200–280 nm), UV-B (280–315 nm) and UV-A (315–400 nm) with only UV-B and UV-A reaching the earth’s surface. Radiation with wavelengths below 290 nm declines to undetectable levels . AlthougLand plants, fungi, and bacteria have several fundamental characteristics in common. While growing, they have minimal possibilities for rapid long-distance displacement and therefore they usually adapt their growth and metabolism to incoming light with functionally overlapping pigments. Plants and microorganisms most often occur in the same terrestrial ecosystems with an equivalent natural climatic environment. The plant phyllosphere gets exposed to UV radiation but the optical properties – as determined by the architecture – of plants, fungi and bacteria differ. Plants have structurally organized multilayer tissues throughout their life cycle, fungi usually have mono-layered mycelia (with fruiting bodies as multi-layered exceptions), and bacteria exhibit significantly less complex organization, as single cells or biofilms. Hence the effect and impact of UV radiation on these organisms will be fundamentally different, ranging from biomolecule damage to secondary metabolite production . InteresHere we review how plants, fungi and bacteria perceive and respond to UV with focus on plants and plant-microbe interactions. An overview of their response mechanisms is presented, and the consequences for their growth and survival are discussed and summarized in Living organisms developed photoreceptors in order to respond to light and optimize growth. Photoreceptors absorb specific wavelengths of radiation, triggering a cascade of events, leading to biological responses. Here we illustrate how plants and microorganisms perceive UV and summarize subsequent downstream responses. Plants have a wide set of photoreceptors, each of which has a specific absorption spectrum. Some of the known photoreceptors absorb in the ultraviolet region. UV responses are ascribed to the sensing by cryptochromes (CRYs) , phototrPhycomyces photobiology has received a lot of attention, including its responses in the UV-range. While positive phototropism is reported for UV-A, Phycomyces bends away from UV-B and UV-C. These opposite responses are genetically independent  and flavonol synthase (FLS1) and chalcone flavanone isomerase (CHI). The UVR8 dependent upregulation of flavonoid biosynthesis, mainly serves to create a sunscreen and thus contributes to the protection of macromolecules against UV-B , supporting the existence of a UVR8-independent pathway in UV-B signaling. Furthermore, this regulation is also independent of PHOT, CRY, phytochromes A, B, D, COP1, and HY5/HYH  and F. fujikuroi (formation of macroconidia). Moreover, CRY-DASH in Neurospora crassa influences light-regulated transcription and is necessary for phase entrainment of the circadian clock  photolyase represses sexual development  induces conidiation. However, the mechanism behind this induction remains elusive and seems to be independent from B. cinerea CRY receptors. In contrast, BcCRY2 even acts as a repressor of conidiation  is responsible for positive phototropism in blue light and UV-A , homologs of WC1 and WC2, are required for photoinduction of conidiation and gene expression regulated by blue light , a complex formed by WC1 and WC2 and named after the original mutation found in rception . As thisrception . WC-likerception , indicatand UV-A . Mechaniand UV-A . Additioue light . Apart fue light .Ralstonia, Pseudomonas and Xanthomonas have LOV and/or BLUF proteins   . On the 1O2) , causing–2s–1), which are sufficient to stimulate photomorphogenic and protective responses  is predominantly controlled by UV-B in a UVR8 dependent manner, but also by UV-A and blue light in a CRY-dependent way  ortholog, and a MUS-18 ortholog (Bcin01g08960).Among plant pathogens, the photodamage responses in ttention . Like otonal WCC . Furtheronal WCC . Apart f. crassa  and T. aroviride , fungi aroviride . The resutations . Of partre found . The firre found . These ere found . BecausePseudomonas sp. have an additional method for DNA repair called mutagenic DNA repair, which helps in the protection against UV-B , flavonoids (plants), mycosporines (fungi) and mycosporine-like amino acids  . The accArabidopsis seedlings  expression , with noHelicobacter pylori activity, namely violaxanthin, luteoxanthin, neoxanthin, antheraxanthin, and lutein , brassinosteroids and abscisic acid, some of which are discussed below. Carotenoids are tetraterpenes that function as accessory pigments during photosynthesis and protect against photooxidation. Carotenoids are strongly induced upon UV exposure. Several plant induced carotenoids were ascribed potent anti-d lutein . Some ted lutein . Recentld lutein , QH6 cond lutein . Moreoved lutein . Antimicgeraniol . Many ofgeraniol . Anothergeraniol , with nogeraniol . Artemisgeraniol  and antigeraniol .Cochliobolus heterostrophus, a pathogen of maize, was for example unable to survive in the field  or mustard oil glycosides break down to release defensive substances. GS are generally found in the Brassicaceae and related families. GS are located within the vacuoles and are physically separated from β-thioglucosidase enzymes (myrosinases). Upon disruption of the plant tissue, myrosinases and GS get in contact and hydrolysis occurs, generating sulfate, glucose and an aglycone moiety. The aglycone moiety is unstable and is converted in either thiocyanates (TCs), nitriles and isothiocyanates (ITCs) . GS and mpestris . The modmpestris .Aspergillus flavus and Penicillium verrucosum, respectively , colorless, water-soluble compounds composed of a cycloheximine or cyclohexenone chromophore which is conjugated with a nitrogen or imino alcohol substituent . Mycospoectively .Arabidopsis  are suppressed by exposing cucumber plants daily for 10 min to UV-B. Interestingly, the observed suppression effect is most significant when only UV-B is used with no background light and when UV-B is combined with red light exposure. The effect was least significant when UV-A or blue light was administered together with UV-B. Given that no additional suppression was observed when treating plants to UV-B, prior to inoculation, the observed effect is mainly directly on the pathogen, rather than induced plant resistance (P. syringae does both), because of poor penetration of UV-C radiation in plant tissues. The practical implementations in greenhouses and in the field are still under development or refinement . Also fltivated .Perhaps the most promising for future applications changing plant-microbe interactions is the modulation of UV-B in the spectrum of the plant environment. The control of UVR8 in plants, and the associated specialized metabolism, combined with detrimental effects of UV-B radiation in microorganisms could provide a part of a steering mechanism for optimal plant growth and defense, which would rely on the capability of the plant of adapting much better to UV-B than the microorganisms. With the quickly developing UV-emitting LED light market, it is ever more possible to apply UV treatments to horticultural crops for various reasons. One major advantage of using UV-B supplementation is that the “treatment” can be instantaneously terminated, in contrast to the lasting impact of chemical plant growth regulators once taken up by the plant. However, the positive effects of UV to induce plant resistance should be taken with caution since some of the induced specialized metabolites which ward off microorganisms and insects may also be undesirable for human consumption, e.g., bitter taste.We can thus conclude that UV-A is mainly neutral or even beneficial for the microorganisms, including the colonization of plants by plant pathogens, for DNA repair and processes associated with efficient infection. By contrast, UV-B and UV-C exposure leads to detrimental effects, often resulting in the death of the microbe and thus interesting for application in horticulture, provided the plant or beneficial organisms are damaged less by UV than its pathogen. Besides these detrimental effects on microorganisms, the upregulation of specialized metabolites in plants and microorganisms can affect the plant-microbe interactions in various ways. The list of advantages versus disadvantages on the use of different UV treatments in horticulture needs to be evaluated depending on a species and purpose-specific basis.FV and LV conceived the project. All authors wrote the article and contributed to scientific discussion.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. The handling editor declared a past co-authorship with one of the authors FV.


Many immuno-therapeutic strategies are currently being developed to fight cancer. In this scenario, oncolytic adenoviruses (Onc.Ads) have an interesting role for their peculiar tumor selectivity, safety, and transgene-delivery capability. The major strength of the Onc.Ads is the extraordinary immunogenicity that leads to a strong T-cell response, which, together with the possibility of the delivery of a therapeutic transgene, could be more effective than current strategies. In this review, we travel in the adenovirus (Ads) and Onc.Ads world, focusing on a variety of strategies that can enhance Onc.Ads antitumoral efficacy, passing through tumor microenvironment modulation. Onc.Ads-based therapeutic strategies constitute additional weapons in the fight against cancer and appear to potentiate conventional and immune checkpoint inhibitors (ICIs)-based therapies leading to a promising scenario. The history of adenoviruses (Ads) begins in the 20th century when the first Ad was discovered and characterized . Over thMastadenovirus. There are 51 human Ad serotypes originally classified based on their ability to be neutralized by specific animal antisera. These can be further subdivided into seven species—or subgroups—(A to G), with a further subdivision of species B into subspecies B1 and B2 on the basis of their capacity to clump erythrocytes of humans, rats and monkeys as well as on the basis of their oncogenicity in rodents. More than 30 simian adenoviruses (sAds) display sequence identities to their human counterparts to such an extent that they have also been included in the taxonomy of human adenoviruses, within species B, C, E, and G [Human adenoviruses (hAds) are nonenveloped viruses with a diameter of 70 to 100 nm. The external protein shell of the virus is icosahedral, with 20 triangular faces, 30 edges, and 12 vertices, and this symmetry is made up in large parts by the major virus protein, hexon. hAds are members of the family Adenoviridae and are classified into the genus E, and G . hAds weE, and G . hAds inE, and G .Ads are usually modified in specific regions , such asAlthough Ads have been originally developed for transgene delivery in gene therapy applications, they have become interesting candidates as an anticancer agent. In fact, the development of replicative Onc.Ads to selectively replicate in cancer cells has been one of the early innovative strategies for cancer gene therapy . ExpressThe main feature of Onc.Ads that leads to their antitumoral effects consists in the ability to induce an immune response against cancer cells in different ways, both modulating the tumor microenvironment (TME) and through the release of tumor-associated antigens (TAAs) and neoantigens that are subsequently processed principally by antigen-presenting cells (APCs). In fact, Onc.Ads induce different types of immunogenic cell death (ICD), such as necrosis, necroptosis, pyroptosis, autophagic cell death, and immunogenic apoptosis. Although Onc.Ads have a significant effect on tumor cells’ growth and tumor size in vivo, they are usually unable to eradicate the whole tumor. In order to potentiate Onc.Ads effects, a local co-administration with HD-Ad vectors expressing a variety of antitumoral proteins revealed to be a promising strategy . Indeed,Since Rudolf Virchow discovered the presence of leukocytes in neoplastic tissues and proposed the link between chronic inflammation and tumorigenesis , a comprAs previously mentioned, the main advantage of using oncolytic viruses (OVs) is their ability to modulate the TME rendering it less immunosuppressive . OVs canThe current scientific trend is to attempt an increase of the antitumoral immune response taking advantages of different agents that can counteract cancer immune escape. Although OVs are able to induce anti-cancer immunity by multiple mechanisms, as described in the previous section, recent updates of clinical trials involving OVs confirm their modest activity as a monotherapy. This can be explained by the inability to optimally infect cancer cells due to (i) neutralizing antibodies, (ii) other antiviral clearance mechanisms, (iii) physical barriers that prevents OVs to reach their entry receptors, or due to viral intrinsic factors such as (iv) engineered cancer-selectivity or transgene expression that can reduce viral fitness and (v) expression of potent transgene(s) that may result in a significant immune response with premature clearance of the OV . In thisUsing immunostimulatory cytokines has become an increasingly promising approach in cancer immunotherapy because they indirectly activate tumor-specific T-lymphocytes capable of rejecting tumor cells from patients with a low tumor burden or because they protect patients from a recurrence of the disease. One of the most promising cytokines for arming oncolytic viruses is granulocyte macrophage colony stimulating factor (GM-CSF) ,36. Its Originally characterized as a potent inducer of natural killer  cell cyt+ CTL priming by signaling through CD40 [+ T-cells, B-cells, and NK-cells as well as memory CD8+ T-cells [OVs have been shown to exert beneficial immunologic responses, including induction of anti-tumor T-cells and modulation of the tumor microenvironment from Th2 to Th1, which has been suggested to contribute to breakage of tolerance in tumors ,48,49. Nugh CD40 . In addi T-cells . Many OV T-cells ,54,55,56 T-cells ,57,58 se T-cells . Further+ T-cells in two mouse glioma models [The tumor necrosis factor receptor superfamily, member 4 (TNFRSF4), also known as CD134 and OX40 receptor, is another member of the TNFR-superfamily of receptors that have gained interest as therapeutic target molecules for cancer immunotherapy. OX40 is not constitutively expressed on resting naïve T-cells and plays a key role in the survival and homeostasis of effector and memory T-cells, and it regulates the differentiation and function of Foxp3+ Tregs . H. Jiana models . In the a models . Therapea models . These dCD40L has also been evaluated in combination with an additional co-stimulatory molecule, named 4-1BBL, to arm OVs . 4-1BBL BiTE molecules are a novel class of immunotherapeutic agents that can activate T-cells independently of MHC expression to lyse target cells. One arm of the BiTE molecule binds CD3-epsilon on the T-cell receptor, whereas the other arm can bind a defined target antigen. Binding of both arms to their corresponding target antigens triggers T-cell activation leading to target cell lysis by apoptosis . Recentl+ T cells, leading to an effective IFNγ-mediated antitumor immune response and a long-term cure of glioma-bearing mice [The recent successes of anti-tumoral immunotherapy approaches, such as ICIs and chimeric antigen receptor T-cell (CAR-T) therapy, have revolutionized cancer treatment, improving efficacy and extending treatment to a larger proportion of cancer patients . Howevering mice . The virIn the process of writing this review, we searched for OVs combined with ICIs clinical trials on the clinicaltrials.gov website; as shown in To date, immunotherapy confirms its powerful efficacy against cancer because it can be long-lasting because of the generation and maintenance of tumor-specific memory T-cells . Among tConsidering the Onc.Ads in the larger set of Ad vectors, we can surprisingly appreciate their utility during the COVID-19 pandemic. A recent work has been published about an Ad5-vectored COVID-19 vaccine that is tolerable and immunogenic at 28 days post-vaccination, even if these findings need further investigation .Cancer cell metabolism is strictly regulated by the TME and cells tend to use aerobic glycolysis to support biosynthetic pathways even in the presence of oxygen . Like tuA novel component in the TME that alters cancer cell metabolism is the microbiome . EvidencTaken together, we think that one of the next goals of viro-immunotherapy could be to develop new strategies useful for a metabolic reprogramming, in order to shape the TME for an effective antitumor T-cell immunity. Understanding metabolic communication between tumor cells and other cellular components  within the TME could have great therapeutic value, and targeting the metabolic crosstalk may directly affect the biological activity of tumor cells and immune cells. In addition, considering the microbiome effects on tumor cell metabolism, we believe that identifying the bacteria or bacterial-derived metabolites connected with a metabolic stress condition within the TME could contribute to the design of novel OVs.


New-onset atrial fibrillation complicating acute myocardial infarction represents an important challenge, with prognostic significance.To study the incidence, impact on therapy and mortality, and to identify predictors of development of new-onset atrial fibrillation during hospital stay for ST-segment elevation myocardial infarction.We studied all patients with ST-elevation myocardial infarction included consecutively, between 2010 and 2017, in a Portuguese national registry and compared two groups: 1 - no atrial fibrillation and 2 - new-onset atrial fibrillation. We adjusted a logistic regression model data analysis to assess the impact of new-onset atrial fibrillation on in-hospital mortality and to identify independent predictors of its development. A p value < 0.05 was considered significant.We studied 6325 patients, and new-onset atrial fibrillation was found in 365 (5.8%). Reperfusion was successfully accomplished in both groups with no difference regarding type of reperfusion. In group 2, therapy with beta-blockers and angiotensin-conversion enzyme (ACE) inhibitors/angiotensin receptor blockers (ARBs) was less frequent, 20.6% received anticoagulation at discharge and 16.1% were on triple therapy. New-onset atrial fibrillation was associated with more in-hospital complications and mortality. However, it was not found as an independent predictor of in-hospital mortality. We identified age, prior stroke, inferior myocardial infarction and complete atrioventricular block as independent predictors of new-onset atrial fibrillation.New-onset atrial fibrillation remains a frequent complication of myocardial infarction and is associated with higher rate of complications and in-hospital mortality. Age, prior stroke, inferior myocardial infarction and complete atrioventricular block were independent predictors of new onset atrial fibrillation. Only 36.7% of the patients received anticoagulation at discharge. The etiology of AF in this context, as in critical illnesses, is most likely multifactorial and includes ischemia, reduced atrial perfusion, glycolytic anaerobic pathways, inflammation, neurohumoral factors, autonomic regulation abnormalities, high left ventricular end-diastolic pressure and elevated atrial pressure.4 The bulk of evidence demonstrates that AF in patients hospitalized for AMI represents an important challenge, concerning the role of antiarrhythmic drugs and antithrombotic treatment.2 The development of AF is linked to poorer prognosis and adverse impact on in-hospital and long-term mortality.6 This seems to also apply for transient AF, which has reversed back to sinus rhythm at the time of discharge.Atrial fibrillation (AF) is the most commonly encountered arrhythmia and often complicates acute myocardial infarction (AMI) with new-onset atrial fibrillation of 6-21%.2With advancements in reperfusion strategies and contemporary treatment with angiotensin-conversion enzyme (ACE) inhibitors, statins and antiplatelet therapy, it could be anticipated that there would be a change in incidence and prognostic impact of AF in the setting of AMI.The purpose of this study was to determine the incidence of AF in patients hospitalized for ST-segment elevation myocardial infarction (STEMI), identify predictors of AF development and analyze its impact on therapy and in-hospital mortality, in the setting of AMI managed according to the most recent guideline strategies.We performed a multicenter retrospective cohort study and identified all hospital admissions from October 2010 to August 2017 with primary discharge diagnosis of ST-segment elevation myocardial infarction included consecutively in a national multicenter Portuguese registry on Acute Coronary Syndromes (ProACS). This Portuguese registry includes 25 cardiology departments and has received the approval and authorization from the National Committee of Data Protection (authorization number 3140/2010) and is registered at ClinicalTrials.gov with identification number NCT 01642329.We used the third universal definition of STEMI as a new ST-segment elevation at the J point ≥ 0.1 mV in two contiguous leads or new left bundle branch block, for over 30 minutes, in a clinical setting consistent with acute myocardial ischemia.New-onset AF was defined as first occurrence of AF at the time of the myocardial infarction (MI) or any time after MI onset, in the absence of prior diagnosis.We divided the patients into 2 groups: Group 1 - Patients who did not develop AF during in-hospital stay for STEMI and Group 2 - Patients with new-onset AF during in-hospital stay for STEMI. We excluded patients with previous diagnosis of AF or previous anticoagulation therapy.In each patient, baseline clinical characteristics, including demographics and patient history (cardiovascular and non-cardiovascular comorbidities) were collected. Data relating to coronary angiography, percutaneous coronary intervention  left ventricular function  and in-hospital and post-discharge medical therapy was also analyzed. The outcome variables studied were in-hospital mortality and complications, namely: heart failure, cardiogenic shock, stroke, major bleeding, atrioventricular (AV) block, ventricular tachycardia, cardiac arrest, mechanical complications of AMI and need for invasive mechanical ventilation, Swan-Ganz catheter, transvenous cardiac pacing, intra-aortic balloon pump or ventricular assist device.We compared the defined outcomes in both groups and performed multivariate data analysis to assess independent predictors for new-onset AF and the impact of new-onset AF on in-hospital mortality.The study protocol is in accordance with the Declaration of Helsinki.Categorical variables were characterized by absolute and relative frequencies, and numerical variables by means and standard deviations for symmetric distributions; for asymmetric distributions, medians and interquartile intervals were used. Normality was assessed by the Kolmogorov-Smirnov test.U test was used to compare the medians.Comparisons between both groups regarding categorical variables were conducted using the chi-square Test or Fisher’s exact Test. For the continuous variables, unpaired t-tests were used to compare the means whenever possible, otherwise, the Mann-Whitney Predictors were determined by adjusting the logistic regression model. Variables were selected to be included in the model using the stepwise (forward) method, together with the likelihood-ratio test. For each variable included in the regression model, adjusted odds ratio and 95% confidence interval (CI95%) were also estimated. The quality of the adjustment of the models was assessed by determining the area under the Receiver Operating Characteristic curve (AUC) and its sensitivity and specificity.A Kaplan-Meier estimate was obtained for time to death. Comparison of two survival functions was conducted by log-rank test.® at a 5% significance level for hypothesis-testing.Statistical analyses were conducted using SPSS 19.0A total of 6957 patients with primary diagnosis of ST-segment elevation myocardial infarction were identified. We excluded 632 patients due to previous diagnosis of AF, previous anticoagulation therapy or missing information. A total of 6325 patients were enrolled . New-onsRegarding patient demographics and comorbidities, patients who developed new-onset AF were older, with higher prevalence of female sex, higher prevalence of hypertension, diabetes mellitus, prior coronary artery bypass grafting (CABG), valvular heart disease, prior heart failure (HF), prior stroke, peripheral arterial disease, chronic obstructive pulmonary disease (COPD) and dementia. There was lower prevalence of smoking habits and family history of coronary disease in group 2. No difference between both groups was found regarding the other characteristics analyzed.Clinical data at admission is shown in Regarding therapy, reperfusion was successfully accomplished in both groups , with noMedical therapy during in-hospital stay and at discharge are shown in New-onset AF was associated with higher rate of in-hospital complications, namely: worse left ventricular function, higher prevalence of HF, cardiogenic shock, AV block, ventricular tachycardia, cardiac arrest, mechanical complications, stroke and major bleeding .It was also associated to a more frequent need of Swan-Ganz catheter, intra-aortic balloon pump, invasive mechanical ventilation and transvenous cardiac pacing . The durWe identified age, prior stroke, inferior located STEMI and complete AV block as independent predictors of new-onset AF .These population-based data reflect the comprehensive experience of a community in a “real-life” setting and is more representative of those treated in clinical practice than populations enrolled in randomized controlled trials.7 However, this number is lower than most older studies, suggesting that despite an increasingly older population with higher prevalence of comorbidities, the incidence of AF in the setting of acute MI has declined.9In our population, new-onset AF was found in 5.8% of patients hospitalized for STEMI, which is in line with recent published data.2DS2-VASc scores. According to Schmitt et al.,1 the use of interventional coronary revascularization has been associated with a notable decline in the AF incidence.1 However, one of the major findings of this study is that new-onset AF complicating STEMI is independent of reperfusion strategy. Reperfusion was successfully accomplished in both groups with no difference regarding the type of reperfusion and no difference in the number of affected or treated vessels between both groups. This is consistent with the OACIS study, which states that AF complicating the clinical course of MI appears to be irrespective of treatment by thrombolytics or PCI.11 However, there seems to be a favorable effect of early reperfusion therapy on preventing new-onset AF, as it also was suggested by a previous study.8 The use of thrombectomy devices, which may suggest a cardioembolic cause, were more frequently used in patients who developed AF, raising the question that some patients with new-onset AF may have had undiagnosed paroxysmal AF.12New-onset AF was found more frequently in older patients, female sex, with higher prevalence of comorbidities, including hypertension, diabetes, previous stroke and previous heart failure and, therefore, higher CHA13 In our population, two-thirds of the patients who developed new onset AF were started on amiodarone.There is insufficient data indicating preferences for rate or rhythm control in AF complicating STEMI. Besides beta-blockers, antiarrhythmic drug use is usually limited to amiodarone.14 As shown in 15 Seventeen percent of patients in group 2 were on P2Y12 inhibitors other than clopidogrel and none of these were on anticoagulants at discharge. It is noteworthy that 6.1% of patients on aspirin during in-hospital stay were not on aspirin when discharged, suggesting that a WOEST trial-like strategy was chosen, which is also supported by a meta-analysis that suggests that a combination of vitamin K antagonists and single antiplatelet therapy is the best choice for AF patients undergoing percutaneous coronary intervention (PCI) considering both efficacy and safety.17 Stenestrand et al.18 and Lopes et al.15 showed that only a small percentage of patients with AF and acute MI received oral anticoagulation therapy at discharge . Our results show a lower rate of anticoagulation: only 20.6% of patients diagnosed with AF during in-hospital stay received oral anticoagulation at discharge. Forty-five percent of patients on anticoagulation at discharge were on NOACs.Selection of antithrombotic therapy was at the discretion of the attending physician and reflects a wide range of therapeutic strategies in a “real-life” setting. Considering the most recent guidelines published by the European Society of Cardiology, the recommendation is that STEMI patients with an indication for oral anticoagulation, as in AF, should be considered for triple therapy with aspirin, clopidogrel and oral anticoagulant for 1-6 months.20 No difference was found regarding statin therapy, unlike what was suggested in previous studies.21Therapy with beta-blockers and angiotensin-conversion enzyme (ACE) inhibitors/angiotensin receptor blockers (ARBs) was less frequently found in patients who developed new-onset AF and although the higher incidence of complications may have limited therapeutic options, this could suggest that renin-angiotensin-aldosterone system inhibitors and beta-blockers may have a protective role, which is in line with previous studies.22 Studies on critically ill patients also suggest new-onset AF is an indicator of clinical severity and poorer prognosis.25New-onset AF was associated with higher incidence of complications, namely stroke, heart failure, cardiogenic shock, arrhythmias, cardiac arrest, mechanical complications and major bleeding, and in-hospital mortality, but was not found as an independent predictor of mortality, suggesting that new-onset AF complicating STEMI may simply be an indicator of poor overall clinical status, which is consistent with previous studies.Age, prior stroke, inferior STEMI and complete AV block were found as independent predictors of new-onset AF.1 of 20 publications also found that new-onset AF is more likely to complicate AMI in older patients.1 Anatomic atrial abnormalities with aging, such as atrial fibrosis and smooth muscle cell proliferation, may provide anatomic substrate for the multiple wavelet re-entry mechanism of AF.26Age and prior stroke reflect patients with more comorbidities. A systematic review by Schmitt et al.27 found that all patients that developed supraventricular arrhythmia had inferior STEMI and right and left atrial ischemia, and concluded that ischemia of the sinus node due to coronary occlusion proximal to the origin of the sinus node artery was probably one of the underlying causes of atrial fibrillation.27 According to Tjandrawidjaja et al.,28 the compromised anatomy of the principal coronary atrial branches is associated with the development of atrial arrhythmia.28Logistic regression analysis showed that inferior STEMI was an independent predictor of new-onset AF in our population. Kyriakidis et al.26 An association between complete AV block and incidence of AF has also been reported previously, particularly in AH type of AV block, due to lesions observed in the atrial muscles including the internodal tracts.29The GUSTO III study found that the development of AF could be due to other post-AMI complications occurring before AF, such as complete AV block.3 In our population, the therapeutic strategy seems to have been based on the notion that new-onset AF in STEMI could represent a transient complication of acute coronary syndrome, as suggested by some studies that have shown that in patients with coronary artery disease, atrial ischemia independently promotes the formation of a substrate for AF and that atrial impulse conduction abnormalities were abolished after reperfusion was achieved with either thrombolysis or primary percutaneous coronary intervention (PCI).31The main question of long-term management of new-onset AF in STEMI and whether to start anticoagulation therapy in these patients remains unanswered, as there is not enough data. In our population, only one in five patients was discharged on oral anticoagulation. Follow-up data on these patients is needed to assess long-term complications. According to some studies, new-onset AF in the acute setting may indicate a propensity to develop arrhythmia again, potentially because of associated comorbidities or other predisposing factors.Although data have been prospectively acquired, the main limitations of this study derive from its retrospective design, which limits some available data.Another limitation is that some patients diagnosed with new-onset AF may in fact have previous yet undiagnosed paroxysmal AF, which may lead to an overestimated incidence. These patients may also represent a different subtype of patients, compared to actual new-onset AF due to the ischemic event.This study reflects a very large registry of new-onset AF in patients hospitalized for STEMI. It shows that in a modern revascularization era, AF is still a frequent complication of acute myocardial infarction. It can be reduced by early reperfusion independently of reperfusion strategy. These registry data confirm the high mortality and morbidity associated with AF, which may suggest the importance of identifying this subgroup of STEMI patients. Age, prior stroke, inferior STEMI and complete AV block were found as independent predictors of new-onset AF in the setting of STEMI. Furthermore, data indicates that only a minority of patients receive oral anticoagulants at discharge. Although the incidence has decreased, we can still expect AF to remain a frequent and worrisome complication of AMI. Further studies answering questions such as how to reduce the risk of AF development, whether the use of new generation drug-eluting stents and modern antithrombotic strategies aimed at better reperfusion will reduce the incidence of new-onset AF during AMI and how to determine the optimal use of oral anticoagulation therapy in combination with antiplatelet therapy in this setting.


These data may provide a reference for incorporating additional core stability exercises in the warm-up routine of physical education classes in school-aged children in the future.Whether the implementation of feasible, equipment-free, and simple core exercises in warm-up routines in physical education classes for school-aged children is beneficial remains unclear. Therefore, this study investigated the effects of a core conditioning in the warm-up routine of physical education classes on trunk muscular endurance, movement capability, and flexibility in this population. In these pre- and post-test control group experiments, 52 healthy, school-aged children (aged 10–11 years) were cluster randomized allocated to either the dynamic core exercise (DCE) group or general physical education (GPE) group. The DCE group performed a 10-min core exercise routine twice per week for six consecutive weeks; the GPE group performed traditional physical education warm-up exercises regularly. The children were assessed by conducting the trunk muscular endurance test , functional movement screen (FMS), and single-leg balance test before and after the intervention. At the end of the intervention, the DCE group demonstrated a significant effect on trunk muscular endurance, movement capability , flexibility, and balance . Furthermore, the DCE group showed significant improvements in all outcome measurements compared with the GPE group ( Numerous studies have highlighted that exercises, particularly in physical education classes, in school-aged children have a key effect on their development ,2,3. BefIn the human body, core muscles include global axial stabilizers, such as rectus abdominis, external oblique abdominis, erector spinae, gluteus, and local axial stabilizers, such as multifidus, transverse abdominis, and internal oblique abdominis . These tp = 0.006) in children aged 8–11 years . Data were assessed visually and statistically for normality  compared with the baseline data. The effect sizes (Cohen’s d) were 0.79 for the dynamic curl-up test, 0.97 for the static curl-up test, 0.91 for the plank test, 1.15 for the side plank test, 1.3 for the FMS total scores, 0.38 for flexibility test, and 1.05 for balance test. The results demonstrated that the DCE group had a greater effect on all muscular fitness test items, FMS total score, and balance tests. The GPE group had significant improvements in the side plank test (p = 0.001), FMS total scores (p < 0.001), and flexibility test (p = 0.012). Conversely, the GPE group had significantly decreased values in the static curl-up (p < 0.009) and plank (p = 0.001) test when compared with the baseline data. Furthermore, the post hoc power analysis was performed using effect size between time points on each parameter   .To the best of our knowledge, this is the first study to investigate the effects of performing dynamic core routines in physical education classes on trunk muscular endurance, movement capability, flexibility, and balance in school-aged children. Our results indicated that a six-week DCE program resulted in significantly superior improvements on all muscular fitness test items, FMS total scores, flexibility, and balance tests. Furthermore, the overall interventional effectiveness was significantly higher in the DCE group than in the GPE group, thus suggesting that the DCE intervention has a greater effect.The core stability affects the effective use of the strength, power, and endurance required. A weak core is a fundamental problem of inefficient movements that ultimately results in injuries .In contrast to previous studies on core strength training programs conducted among pediatric age groups, we performed this experimental control study on school-aged children and required minimal time and no equipment to enhance outcomes efficiently. Ozmen et al. found that a six-week core strength training program significantly improved balance and core endurance . Howeverp < 0.001, effect size: 0.38–1.3), indicating enhancement of core stability. However, we cannot rule out the possibility that the measurements were influenced by learning effects associated with the involved movements . Furthermore, our findings demonstrated that the FMS scores and single-leg balance test results were considerably better following DCE . Furthermore, the DCE group showed significant improvements with moderate-to-large effects in movement capability and balance tests compared with the control group . This indicates that children can increase their core strength and endurance over time, enabling them to maintain postural control, reduce inefficiency in their movement patterns [The dynamic core stability intervention can be beneficial from the implementation of dynamic whole-body movements rather than trunk muscles alone. Additionally, such a moderate intensity warm-up can engage the major muscles for more vigorous movements by increasing the core body temperature and motor unit recruitment . In the patterns , and imppatterns ,40.Interestingly, the GPE group displayed improvements in the sit-and-reach test, lateral plank test, and FMS total scores. In contrast to previous studies, our study included a control group  to verify the effects of the intervention. Our data indicated that including only a GPE warm-up routine may be suboptimal for improving physical fitness in school-aged children. Furthermore, in this study, the GPE group performed routine jogging, light intensity of general active range-of-motion exercises, and static stretching for upper and lower extremities. We postulated that these may result in unanticipated adaptations to one’s flexibility and core stability. Therefore, this is not to say that the general warm-up protocol should be completely replaced by dynamic core stability exercises in a child’s pre-event program but rather that physical education teachers should consider the potential additional effects on exercise performance.For practical application, we recommend that school-aged children begin their exercise program with a 10-min warm-up DCE routine (twice a week for six weeks), which requires minimal time and equipment to complete. This program is designed to train the major trunk and lower extremity muscles necessary for motor competence and functional performance. This program can potentially be adapted to children of other age groups and may considerably improve children’s physical fitness, which is particularly imperative given the global crisis of pediatric obesity.A six-week DCE program as a warm-up exercise significantly improved trunk muscular endurance, movement capability, flexibility, and balance in school-aged children. These results may provide a reference for the incorporation of core stability exercises as part of a warm-up routine for school-aged children in physical education classes. The exercise can feasibly be performed in physical education classes. Future studies should investigate the long-term effects of this program on a larger population.


Our group recently developed a urinary 6-biomarker panel for the diagnosis of prostate cancer (PCa) which has a higher level of accuracy compared to the serum prostate specific antigen (PSA) test. Herein, urine from an independent cohort of PCa patients and cancer-free controls was analyzed to further validate the discriminative power of that panel. Additionally, urine from patients diagnosed with bladder cancer (BC) and renal cancer (RC) were included to evaluate the site-specificity of the panel. Results confirmed the ability of the 6-biomarker panel to discriminate PCa patients from controls, but not from other urological cancers. To overcome this limitation, an untargeted approach was performed to unveil discriminant metabolites among the three cancer types. A 10-biomarker panel comprising the original panel plus four new metabolites was established to discriminate PCa from controls, BC, and RC, with 76% sensitivity, 90% specificity, and 92% accuracy. This improved panel also disclosed better accuracy than serum PSA test and provides the basis for a new non-invasive early detection tool for PCa. Globally, prostate cancer (PCa) ranks first amongst all male urological cancers and second in incidence of all cancers in men . Due to Metabolic rewiring has recently been recognized as a hallmark of cancer cells , boostinA targeted metabolomics approach a was firThe untargeted approach consisted on the comparison of the urinary volatile profiles (VOCs and volatile carbonyl compounds (VCCs)) of PCa patients with that of BC and RC patients. Results disclosed a good separation of PCa from BC and RC in the PLS-DA models . OverallThe seven candidate biomarkers disclosed by untargeted analysis were assessed for their ability to perfect an extended biomarker panel providing the best combination of sensitivity, specificity, and accuracy for PCa detection. Considering a PLS-DA based algorithm, four metabolites were selected, namely ethylbenzene, heptan-2-one, methyl benzoate, and 3-methylbenzaldehyde . TherefoThe major novelty of this study design was the inclusion of patients with other urological cancers to evaluate the performance of the biomarker panel to discriminate not only PCa vs. cancer-free individuals, but also PCa vs. BC and RC  . This crThose results demonstrated that BC or RC patients may be false positives in a PCa screening strategy based on the 6-biomarker panel, constituting a relevant limitation in clinical practice. To overcome this, we performed an untargeted metabolomic study to look for new biomarkers capable of discriminating PCa from the other urological cancers. This approach allowed the improvement of the previously established biomarker panel through the addition of four new volatile compounds, discriminative of PCa vs. BC and RC. Thus, the improved panel included five aldehydes , two ketones , two aromatic hydrocarbons (methyl benzoate and ethylbenzene), and one polycyclic organic compound (dihydroedulan IA). From these, only hexanal ,22,23,24In the last decade, several candidate biomarkers have been proposed using metabolomic approaches, including mainly amino acids and derivatives, organic acids, and sugars . OverallThese results emphasize the potential of volatile biomarkers for development of a non-invasive screening tool for clinical diagnosis. However, the analytical techniques used in metabolomic approaches might not be feasible for real-time diagnostic applications due to several limitations, such as low sample throughput, high costs, and the requirement for trained personnel and sophisticated software. Thus, future research may rely on the translation of the volatile signatures detected by GC-MS analysis to the application of a fast, cheap, and portable e-nose device. In this regard, our defined panel holds the potential for development of an optimized e-nose—with sensor arrays targeting specific PCa-related volatile compounds—for routine clinical use with high accuracy for PCa diagnosis.Urine samples were collected at the Portuguese Oncology Institute of Porto from a total of 80 men, comprising 20 PCa patients, 20 BC patients, 20 RC patients, and 20 cancer-free individuals (controls) . This stPreparation of urine samples and analytical conditions followed the protocol described by Lima et al. . To deteVOCs and VCCs analyses were performed in a 436-GC  coupled to a Bruker Scion SQ MS detector  and a 436-GC  coupled to an EVOQ TQ-MS , respectively. Details on software, GC and MS acquisition parameters, and metabolite identification are described in Lima et al. .t-test  [p-value > 0.05 were removed from the matrix. Finally, the untargeted models were scaled to pareto and the models based in the biomarker panel were scaled to unit variance (UV).Data pre-processing was performed in MZmine 2.18  using thoftware)  and all Multivariate statistical analysis (MVA) included principal component analysis (PCA) to detect trends and possible outliers, followed by partial least squares discriminant analysis (PLS-DA). In addition, the robustness of all PLS-DA models was confirmed through 7-fold cross validation and permutation test in SIMCA-P 15 .t-test with Welch correction test  or unpaired Mann–Whitney U-test  were applied to all metabolites with VIP higher than 1 in the untargeted models (VOCs and VCCs) . In addition, percentage of variation and effect size were computed for all statistically significant metabolites. The metabolites significantly altered in all comparisons  were considered specific for PCa. To select the most important metabolites among these, a PLS-DA algorithm was applied. The selected biomarker panel was used to construct a classification model and the corresponding receiver operating characteristic (ROC) curve using the MetaboAnalyst 4.0 software [Unpaired Student’s software .In conclusion, we propose an improved volatile urinary biomarker panel to simultaneously discriminate PCa from cancer-free subjects and carriers of other common urological cancers, holding the potential for the development of a non-invasive early detection tool for PCa that outperforms serum PSA sensitivity and accuracy. In the future, we plan to extend the PCa-specificity of this novel panel, testing larger cohorts of cancer patients and controls, including common non-urological cancers in males . Furthermore, taking into consideration that benign prostate hyperplasia, urinary tract infection, and prostatitis are important confounding factors in PSA test, we also plan to include urine samples from patients with these diseases.


The present study was conducted to develop a predictive type of PC-SAFT EOS by incorporating the COSMO computations. With the proposed model, the physical adjustable inputs to PC-SAFT EOS were determined from the suggested correlations with dependency to COSMO computation results. Afterwards, we tested the reliability of the proposed predictive PC-SAFT EOS by modeling the solubility data of certain pharmaceutical compounds in pure and mixed solvents and their octanol/water partition coefficients. The obtained RMSE based on logarithmic scale for the predictive PC-SAFT EOS was 1.435 for all of the solubility calculations. The reported values (1.435) had a lower value than RMSE for COSMO-SAC model (4.385), which is the same as that for RMSE for COSMO-RS model (1.412). The standard RMSE for octanol/water partition coefficient of the investigated pharmaceutical compounds was estimated to be 1.515. The Conductor-like Screening Model (COSMO), a continuum solvation model based on quantum chemistry, was firstly introduced by Klamt and Schuurmann2. The remarkable advantages of COSMO calculation in predicting physical and chemical properties were a motivation to expand its applications by employing different software. Several software packages, such as GAMESS3 and Dmol34, were designed for COSMO-based quantum chemistry calculations. Consequently, some thermodynamic models have been developed based on COSMO calculations. The activity coefficient models based on COSMO, such as COSMO-RS7 and COSMO-SAC8, have been known as successful tools in thermodynamic scope. Studies by Tung et al.9, Zhou et al.10, Paese et al.11, Xavier et al.12, Bouillot et al.13, Shu and Lin14, Buggert et al.15, and Mahmoudabadi and Pazuki16 are some examples. Despite the remarkable characteristic of COSMO-based model, the vapor–liquid equilibria in COSMO-based model are computed by ideal gas assumption for vapor phase. Meanwhile, compressibility factor, viscosity, and different chemical and physical properties for various phases could be obtained via the equations derived from equations of state. The above-mentioned discussions have encouraged scientists to explore the remarkable properties of quantum chemistry calculations in combination with other equations of state in two forms: (i) combination with other equations of state (EOS) in the form of contribution in order to increase accuracy and (ii) combination with other equations of state in order to perform a predictive model.Computation of phase equilibria and thermodynamic properties is believed to be one of the significant objects in thermodynamic investigations, which directly relates to molecular structure, functional group, and chemistry of molecule. Quantum chemistry, as an approach to calculating the mechanical properties of molecules, has been extensively applied in thermodynamic computations. Quantum chemistry computation based on 7 and COSMO-SAC8, to other equations of state. Lee and Lin17 considered a combination of COSMO-SAC and Peng-Robinson (PR) for thermodynamic modeling. Pereira et al.18 examined the COSMO-RS model in selecting an association scheme and molecular models for soft-SAFT EOS in the ionic liquids system. Cai et al.19 investigated the PR + COSMO-SAC model for solid solubilities in supercritical carbon dioxide.In method (i), several researchers have examined adding quantum chemistry models, for instance, COSMO-RS20 considered two alternative methods based on COSMO calculations for estimating the PHSCT EOS parameters in order to predict vapor–liquid equilibria (VLE) behavior for refrigerants: (i) PHSCT EOS parameters were fitted by the computed activity coefficients from COSMO-RS model and (ii) PHSCT EOS parameters were obtained directly from quantum calculations. Cassens et al.21 utilized electrostatic properties of pharmaceutical compounds to identify Perturbed-Chain Polar Statistical-Associating Fluid Theory (PCP-SAFT EOS) parameters. The PCP-SAFT pure-component size and shape parameters of solutes were obtained from quantum chemistry by considering available size and shape parameters for PCP-SAFT EOS and introducing some relations between dispersion energy and reduced dipole moment to size and shape parameters. Ferrando et al.22 proposed a new methodology based on Monte Carlo simulations in order to determine the PPC-SAFT EOS-associated scheme and parameters by focusing on 1-alkanol molecules. Van Nhu et al.23 computed PCP-SAFT EOS parameters for 67 compounds using quantum chemical ab initio and density functional theory methods. Van Nhu et al.23 induced a relationship between quantum calculations and pure parameters for PCP-SAFT EOS. They tuned PCP-SAFT EOS parameters with vapor pressure and density data and changed atomic radii inputs to COSMO calculations so that it match with the obtained parameters of PCP-SAFT EOS. Singh et al.24 employed the Gaussian03 software package in performing quantum results. They utilized different theories for obtaining the dispersion energy, the dipole moment, the quadrupole moment, and the dipole–dipole dispersion coefficient. Leonhard et al.26 proposed a novel method for computing PCP-SAFT parameters and introduced a combining rule based on quantum chemistry. They calculated the dispersion energy, the dipole moment, the quadrupole moment, and the dipole–dipole dispersion coefficient with quantum calculations and tuned the remaining parameters by experimental data.In method (ii), several studies have considered quantum chemistry computations combined with other commonly used equations of state toward a new predictive model for thermodynamic computations. In this method, the quantum computations are not directly incorporated in equations of state. The popularly applied equations of state have some adjustable parameters with physical meanings, which are tuned by experimental data. However, in this method, the adjustable physical parameters of EOS are determined based on quantum chemistry computations. There are certain studies in this regard as reported in Table 3 module in Material studios 2017 with the discussed characteristics. The newly performed predictive PC-SAFT EOS was applied for estimating pharmaceutical compound solubilities in various pure and binary solvents. The octanol/water partition coefficients of the considered pharmaceutical compounds were also obtained with the proposed predictive PC-SAFT EOS. They were then compared to the experimental data. Through this modification to PC-SAFT EOS, the domains of applicability of COSMO computations would expand and the adjustable parameters of PC-SAFT EOS could be estimated.According to the literature review, the quantum chemistry was mostly manipulated in PC-SAFT type EOS to perform a predictive model. However, a few studies considered original PC-SAFT EOS and constructed a predictive PC-SAFT EOS using quantum chemistry. Based on the methods implemented for performing predictive EOS, the majority of the studies have relied on the complicated equations and methods to estimate the parameters. Some others utilized the previously tuned parameters to define their equations. There is no study in which COSMO results are directly utilized for setting up their EOS. In this work, the adjustable parameters for PC-SAFT EOS could directly calculate through simple equations with decency to obtain the results based on COSMO calculations. The current study aimed to make a predictive PC-SAFT EOS by incorporating COSMO calculations from the Dmol3:4.In quantum chemistry, molecular properties are determined via several electron time-independent equations3 module in Material studios 2017 software were generated. According to Lin and Sandler 8, GGA (VWN-BP) is considered as a density function, and the accuracy of computation is set by quality fine. In electronic options, multipolar expansion is selected octupole. The calculations are run at four parallel cores. The other options are set to the default values of DMol3.In this study, 35 pharmaceutical compounds and 15 solvents were considered for solubility calculations. Performing the predictive PC-SAFT EOS, we examined 26 non-associating organic compounds, such as alkanes and alkenes, and 13 associating organic compounds, alkanols for instance. For all of the 80 surveyed compounds, the COSMO files from the DmolA) and the total volume of the cavity (V) were the desirable properties gathered for all of the 80 examined compounds. Prior to using the surface area of the cavity and total volume of the cavity, a unit conversion from Bohr to angstrom is required (1 Bohr = 0.52918 Angstrom).Figure 27. In PC-SAFT EOS, the perturbation theory for dispersion forces and hydrogen bonding were incorporated into a hard-chain reference fluid through the following equation:aPC-SAFT) is obtained by the Helmholtz energy contributions from the reference system, hard chain (ahc), dispersion force (adisp), and hydrogen bonding (aassoc) as follows28:i at site A. i in its mixture with hard-sphere contacts. M is the number of associated sites. The details of formulations of the above-mentioned equations were presented for non-associating contributions in Gross and Sadowski27 and for associating contributions in Huang and Radosz29.Perturbed-Chain Statistical-Associating Fluid Theory (PC-SAFT) was firstly developed by Gross and Sadowskim, segment diameter The total five parameters of PC-SAFT EOS for each molecule, segment number i in the mixture and pure states at the same system temperature and pressure, respectively. The fugacity coefficient for component k (z) were estimated using the PC-SAFT EOS compute as follows:In this study, the solubility of pharmaceutical compounds in different solvents were desirable properties determined by knowing the activity coefficient of pharmaceutical compounds (kij) is defined for correcting the segment-segment interactions of unlike chains and calculated with the combining rule below:In order to obtain accurate results for the mixtures, the binary interaction parameter (kij) increases the flexibility of the PC-SAFT EOS along with accurate modeling of more complicated systems.The addition of an extra binary interaction parameter , and water (W) at infinite dilution, is measured with partition coefficient as follows31:Once pharmaceutical compounds are dissolved in two immiscible liquid phases containing octanol and water, the components will be distributed between two phases. The distribution of component i  and the segment diameter (A) and the total volume of the cavity (V) was obtained via the COSMO file. The following two equations could be written based on definitions for the surface area of the cavity32 and the total volume of the cavity27:A) and total volume of the cavity (V)) as follows:By incorporating two equations, the segment diameter (m) for PC-SAFT EOS is obtained as follows:Knowing the segment diameter, the segment number  and the segment diameter  was firstly introduced by Klamti in solvent S  the dissolution of the solute into a perfect conductor and (ii) the restoration of the conductor to the actual solvent. The residual part of the COSMO-SAC was defined as follows36:The conductor-like screening models-segment activity coefficient (COSMO-SAC) was proposed by Lin and Sandler27, we observed that the predictive PC-SAFT EOS has a lower segment number and larger segment diameter. The dispersion energies obtained from Eq. 17 had larger values with respect to dispersion energies reported by Gross and Sadowski27. With the increase in the molecule volume and molar mass, both the segment number and segment diameter were enhanced, which obeys the same trend as by Gross and Sadowski27. The increment of molecule size for each hydrogen bonding site reduced association energies while Gross and Sadowski28 presented various trends for association energy.The Predictive PC-SAFT EOS parameters for all the 35 considered pharmaceutical compounds and 15 solvents were obtained employing Eqs. 37 demonstrated the practically unrealistic and even nonphysical predictions of different version of SAFT EOS, such as PC-SAFT EOS at certain conditions. Privat et al.38 exhibited that the PC-SAFT EOS may have up to five different volume roots, some of which are not realistic. To test the above-mentioned statement for predictive PC-SAFT EOS, we followed the same approach as discussed in Privat et al.38. The Polishuk16. The COSMO-RS results were obtained in this study utilizing COSMOtherm software. Based on Fig. 16. The RMSEs for COSMO-RS model was 1.412. According to Table 23Afterwards, to examine the reliability of the obtained PC-SAFT parameters, experimental solubility data of all the 35 surveyed pharmaceutical compounds in 15 pure solvents were simulated with PC-SAFT EOS. For solubility in the binary system, 918 data for 110 systems over the temperature range of 262–360 K were explored. The mole fractions of the considered solubility data varied from Figure It could be concluded that the observed errors in predictive PC-SAFT were originated from different sources, including (i) various quantum chemistry theories manipulated in COSMO file generation, (ii) inaccuracy in values of atomic radii input for COSMO calculations, (iii) errors from neglecting binary interaction parameters in PC-SAFT EOS, and (iv) deficiencies in original PC-SAFT EOS theories.i. The binary interaction parameters were regressed employing fminsearch algorithm in MATLAB programming software. The regressed binary interaction parameters for dapsone-acetone, dapsone-methyl acetate, and dapsone-methanol were − 0.0455, 0.0156, and 0.0758, respectively. The calculated solubilities of dapsone in acetone, methyl acetate, and methanol are plotted in Fig. For instance, the calculated dapsone solubilities in acetone, methyl acetate, and methanol from predictive PC-SAFT illustrated 50–60 AAD% compared to experimentally reported data. As described before, tuning a binary interaction parameter was suggested for correcting the reported error. Therefore, the binary interaction parameters between dapsone-acetone, dapsone-methyl acetate, and dapsone-methanol were fitted by justifying the experimental data and the following objective function:In addition to pure solvents, experimental solubility data in binary solvents was also tested. For solubility in the ternary system, 453 data for 10 systems over the temperature range of 277–325 K were explored. The mole fractions of the considered solubility data varied from In Fig. Octanol/water partition coefficients for all the surveyed pharmaceutical compounds were calculated by predictive PC-SAFT and compared with the reported experimental data in Table ab initio, Monte Carlo, and COSMO-based models are certain approaches to quantum chemistry calculations.According to online literature, various attempts were made in order to attain a predictive-type PC-SAFT EOS. In predictive-type PC-SAFT EOS, the input parameters to the model are determined from another procedure instead of tuning with experimental data. Group contribution methods and correlation with molecular mass are a few examples. Among these approaches, developing a predictive model based on quantum chemistry calculation has become incredibly popular. The In the current study, quantum chemistry calculations based on COSMO were selected in order to make a predictive-type PC-SAFT EOS. The surface area of the cavity and total volume of the cavity were obtained with the COSMO file of 80 compounds consisting of pharmaceutical compounds, non-associating organic compounds, associating organic compounds, and popular solvents for pharmaceuticals. Two mentioned properties were incorporated into the adjustable input parameters for the PC-SAFT EOS by introducing some correlations based on COSMO results. Thus, a predictive PC-SAFT EOS was introduced utilizing the above-mentioned methods and correlations. To highlight the operability of the newly modified EOS, the solubility data of pharmaceutical compounds in pure/mixed solvents were simulated by the predictive PC-SAFT EOS and compared to COSMO-SAC and COSMO-RS models. According to the results of predictive PC-SAFT EOS, a good agreement was observed between model and experimental data in both binary and ternary systems. The predictive PC-SAFT EOS presented a better approximation of the experimental data when compared to COSMO-SAC model. The predictive PC-SAFT EOS and COSMO-RS model demonstrated the same accuracies. The observed deviations of predictive PC-SAFT from experimental data could be elaborated with the basic theory of applied density function, default values for atomic radii in COSMO calculations, the theory of PC-SAFT EOS, and elimination of binary interaction parameters. As in one case for dapsone, a good representation of experimental data with predictive PC-SAFT was concluded by fitting binary interaction parameters. It shed light to the fact that the volume and surface of molecules could be calculated employing any methods on top of COSMO, which is a crucial feature of the developed predictive PC-SAFT EOS.Supplementary Information


Due to COVD-19, many health/wellness programs transitioned from in-person to virtual. This mixed methods study aims to explore older adults with Parkinson’s disease (PD) perceptions of in-person versus virtual Parkinson’s-specific exercise classes. Attitudes, perceptions, and experiences were determined through focus groups  among older adults with PD and an online survey . Eighteen respondents attended both in-person and virtually . Four respondents only attended in-person, citing reasons such as difficulty with computer access to virtual classes, limited internet, easier accessibility to in-person classes, and physical injury preventing attendance to any classes. Respondents who participated in both delivery methods preferred virtual classes. Time, convenience, comfort at home, and not having to navigate transportation barriers supported participants’ preference for virtual classes. The majority of respondents indicated their fatigue and mental health were either unchanged or improved. Eighty-nine percent reported improved mobility since attending either class; specifically, in balance (n=8), flexibility (n=7), and coordination (n=3). Older adults with PD who attended both classes had minimal difficulty with computer usage and accessing the virtual program with only one participant reporting difficulty transitioning from in-person to virtual classes. Seventy percent stated they would enjoy a combination of on-site and virtual programming. Eighty-nine percent and seventy-seven perfect felt the virtual-based program was safe and beneficial, respectively. Participants who transitioned from an in-person to a virtual exercise program for people with Parkinson’s disease felt the program was safe, effective, and improved or prevented declines in their mobility.


The bone marrow is home to well-balanced normal hematopoiesis, but also the stage of leukemia’s crime. Marrow adipose tissue (MAT) is a unique and versatile component of the bone marrow niche. While the importance of MAT for bone health has long been recognized, its complex role in hematopoiesis has only recently gained attention. In this review article we summarize recent conceptual advances in the field of MAT research and how these developments impact our understanding of MAT regulation of hematopoiesis. Elucidating routes of interaction and regulation between MAT and cells of the hematopoietic system are essential to pinpoint vulnerable processes resulting in malignant transformation. The concept of white adipose tissue contributing to cancer development and progression on the cellular, metabolic, and systemic level is generally accepted. The role of MAT in malignant hematopoiesis, however, is controversial. MAT is very sensitive to changes in the patient’s metabolic status hampering a clear definition of its role in different clinical situations. Here, we discuss future directions for leukemia research in the context of metabolism-induced modifications of MAT and other adipose tissues and how this might impact on leukemia cell survival, proliferation, and antileukemic therapy. Dicer1, the key factor of microRNA maturation in osteoprogenitors, but not in mature osteoblasts, led to disruption of hematopoietic integrity and resulted in myelodysplasia and emergence of acute myelogenous leukemia (AML) in mice [via endocrine routes of interaction.The bone marrow microenvironment (BMM) is characterized by a complex network of different cell types providing key signaling events for the regulation of hematopoiesis. Although leukemia is caused by a series of genetic hits to hematopoietic stem and progenitor cells leading to malignant transformation –3, evide in mice . Further in mice . The che in mice . On thes in mice , and it  in mice . There iWhen talking about AT, we usually refer to the two classical types, i.e., white adipose tissue (WAT) and brown adipose tissue (BAT) Fig. . WAT covnomen est omen, uncouples the oxidative phosphorylation from the ATP-synthase to generate heat instead of ATP [BAT, instead of saving fat, has the unique capability of using up stored triglycerides for production of heat in a process called nonshivering thermogenesis . This prd of ATP . The broBeige  adipocytes can arise within WAT depots upon chronic cold exposure (reviewed in ) Fig. 11. They rThe color yellow was assigned to marrow adipose tissue (MAT) Fig. . In a heThe bone cavity formed by osteoblasts harbors the BM and is home to hematopoiesis Fig.  maintainRecently, the murine BM was analyzed by single-cell RNA-sequencing (scRNA-seq) either under normal or stressed conditions  or in thHSCs critically depend on factors provided by the BMM such as stem cell factor (SCF) and CXCL12 –25. PeriHSCs reside in special “niches” in which their self-renewal and differentiation are influenced by signals from the surrounding BMM, a concept dating back to 1978 . Most HSIn utero and at the time of birth, the bone cavities are predominantly occupied by hematopoietic red marrow . The forThe existence of two different types of MAT was suggested already in 1976 . RecentlFor decades MAT was considered a mere filler of the otherwise empty BM space and we have just begun to understand that its function goes far beyond. Comparable to WAT, MAT was identified as an endocrine organ . BM adipInterestingly, MAT increases under certain clinical conditions including osteoporosis , 45, ageTaken together, MAT is characterized by a dynamic plasticity and region-specific differences. The large variety of influencing factors seem to hamper systematic investigations and may account for controversies found in the literature.We are only beginning to understand the role of MAT in normal hematopoiesis. In a landmark paper, Naveiras et al. identified MAT as negative regulator of hematopoiesis . They shThere is accumulating evidence that MAT also influences survival and proliferation of leukemic blasts. Decline of normal hematopoiesis during leukemia progression is a well-known phenomenon, but has only recently been attributed to a disturbance of MAT function by AML cells resulting in compromised myelo-erythropoiesis . In contLepr expression and activation of the downstream signaling molecule PR/SET domain 1 in leukemic cells [Lepr and might therefore be resistant to fasting-induced upregulation of this pathway. In ALL, fasting not only prevented leukemia development, but also resulted in differentiation and depletion of leukemic cells. Most importantly though, fasting also effectively inhibited the growth of human patient-derived B-ALL in a xenograft model and the expression of LEPR signaling-related genes was correlated to prognosis in pediatric pre-B-ALL [Caloric restriction is known to result in an increase of MAT . Interesic cells . In contre-B-ALL . Clearlyre-B-ALL . The conre-B-ALL ). The fire-B-ALL .Back in 1962, infiltrations of leukemia or lymphoma cells were identified in subcutaneous AT of patients . In miceEvidence is accumulating that leukemic cells, when in close proximity to adipocytes, are protected from chemotherapy-induced cytotoxicity. Mechanisms by which adipocytes might provide shelter to leukemic blasts were intensively studied by Mittelman et al. in both, in vitro and in vivo models –73. Muril-asparaginase (ASNase) [The same syngeneic HFD-ALL mouse model was used for the analysis of the response to (ASNase) , a first(ASNase)  and it i(ASNase) , obesity(ASNase) . Via sec(ASNase) .Most importantly, WAT is not only able to attract leukemia cells via the secretion of chemotactic factors  but seemThe concept of targeting the interaction of leukemia cells with their specific niche in the BM has emerged only recently , 75 and +) cells was analyzed, which contained four different clusters of which two expressed adipogenesis-associated marker genes [+ cells were identified as major source of prohematopoietic factors in the BMM, amongst them SDF-1 (Cxcl12), SCF (Kitl), IL-7 (Il7), IL-15 (Il15), IL-34 (Il-34), M-CSF (Csf1), BMP-4 (Bmp4), CCL-19 (Ccl19), and CCL-2 (Ccl-2) [Recent scRNA-seq studies on murine BM stroma identified factors with specific relevance for leukemia treatment. For example, the compartment of leptin receptor-positive (Leprer genes . Interes (Ccl-2) . Gene ex (Ccl-2) . Several (Ccl-2) ). Whethe (Ccl-2) , the dia (Ccl-2) , mechani (Ccl-2) , and exe (Ccl-2) . One stu (Ccl-2) .Lepr expression in leukemia cells and activation of downstream signaling routes as main cause leading to inhibition of leukemia initiation and progression. Vice versa, attenuated Lepr expression was identified as essential for development and maintenance of ALL [Fasting, a condition known to enhance MAT, inhibited the engraftment and progression of B-ALL and T-ALL in several mouse model systems tested . From the of ALL . All in Besides acting as a member of the local BMM, adipocytes can certainly also interact with leukemia cells via their secreted factors in an endocrine manner. This assumption is underlined by the fact that obesity, which is characterized by the excessive accumulation of WAT, leads to an increased risk of developing and dying from leukemia as recently revealed in a meta-analysis of prospective cohort studies . The facFasting is currently propagated as the holy grail of cancer treatment by some researchers . Since aLepr was already identified as protective mechanism in B-ALL and T-ALL development [It is very obvious that the key alterations induced by fasting are related to altered function of AT. A decrease in leptin and a subsequent upregulation of elopment . It willelopment . How thielopment . RSV inhelopment . At the elopment . It is celopment , reduceselopment  and, of elopment . By the elopment  and leadelopment  and, impelopment .Another important finding that requires appropriate follow-up is the accumulation of leukemia cells in WAT found in several mouse models , 68, 71 Taken together, although neglected in the past, MAT as well as WAT should be considered important and relevant in the context of leukemia. Their secretion products such as growth factors, cytokines, chemokines, matrix molecules, or metabolites have an impact on the function of leukemic cells opening up new therapeutic opportunities to fight hematological malignancy.


A combination of X-ray crystallography, NMR, and mass spectrometry has revealed how diverse small-molecule inhibitors bind Bruton’s tyrosine kinase and alter the conformation of this enzyme. Related research article Joseph RE, Amatya N, Fulton DB, Engen JR, Wales TE, Andreotti A. 2020. Differential impact of BTK active site inhibitors on the conformational state of full-length BTK. eLife9:e60470. doi: 10.7554/eLife.60470The switching of enzymes between active and inactive states, a process known as enzyme regulation, is crucial in cell biology, and the breakdown of the process has been implicated in many diseases. A number of small-molecule inhibitors work by blocking enzyme function, but efforts to evaluate the efficacy of such inhibitors have been hampered by the lack of a detailed understanding of how they work. For example, some small-molecule inhibitors work by making localized changes to the shape of the enzyme at the site where they bind, whereas others work by inducing changes in another part of the enzyme, a phenomenon known as allostery.A clearer picture of allostery requires detailed knowledge of enzyme function and the underlying protein dynamics . Now, inBTK is a kinase that regulates the immune responses of B- and T-cells, and blocking its activity can help suppress inflammatory responses and treat lymphomas and leukemias . High-reWhile X-ray crystallography can provide information on the active and inactive conformations of enzymes at high resolution, additional techniques are needed to understand how enzymes and small-molecule inhibitors interact in solution, so Joseph et al. combined X-ray crystallography with nuclear magnetic resonance (NMR) and amide hydrogen/deuterium exchange mass spectrometry (HDXMS). NMR provides a global overview of conformation, including the transitions between the active and inactive state, while HDXMS localizes conformational changes at a peptide level.The experiments revealed that BTK exists in an ensemble of conformations, encompassing the inactive and active states of the enzyme . Joseph This work underscores the limitations of using structural snapshots from X-ray crystallography or cryo-electron microscopy alone to map small-molecule interaction sites, or to describe allosteric effects. It also highlights how combining NMR and HDXMS with static structural data will lead to more complete descriptions of drug-enzyme interactions. More generally, combining structural, dynamic and computational approaches will help researchers to design inhibitor drugs that are not rendered ineffective by disease-resistant mutations.