Treatment outcome clustering patterns correspond to discrete asthma phenotypes in children

Despite widely and regularly used therapy asthma in children is not fully controlled. Recognizing the complexity of asthma phenotypes and endotypes imposed the concept of precision medicine in asthma treatment. By applying machine learning algorithms assessed with respect to their accuracy in predicting treatment outcome, we have successfully identified 4 distinct clusters in a pediatric asthma cohort with specific treatment outcome patterns according to changes in lung function (FEV1 and MEF50), airway inflammation (FENO) and disease control likely affected by discrete phenotypes at initial disease presentation, differing in the type and level of inflammation, age of onset, comorbidities, certain genetic and other physiologic traits. The smallest and the largest of the 4 clusters- 1 (N = 58) and 3 (N = 138) had better treatment outcomes compared to clusters 2 and 4 and were characterized by more prominent atopic markers and a predominant allelic (A allele) effect for rs37973 in the GLCCI1 gene previously associated with positive treatment outcomes in asthmatics. These patients also had a relatively later onset of disease (6 + yrs). Clusters 2 (N = 87) and 4 (N = 64) had poorer treatment success, but varied in the type of inflammation (predominantly neutrophilic for cluster 4 and likely mixed-type for cluster 2), comorbidities (obesity for cluster 2), level of systemic inflammation (highest hsCRP for cluster 2) and platelet count (lowest for cluster 4). The results of this study emphasize the issues in asthma management due to the overgeneralized approach to the disease, not taking into account specific disease phenotypes

Chemokines during anaphylaxis : the importance of CCL2 and CCL2-dependent chemotactic activity for basophils

Background: The role of chemokines in anaphylaxis is unclear. Methods: We prospectively recruited 49 patients presenting to the emergency department with an acute episode of anaphylaxis and 28 healthy subjects. We measured serum levels of the chemokines CCL2, CCL5, CCL7, CCL8, CCL11, CCL13, CCL17, CCL21, CCL22, CCL24, and CCL26, tryptase, the absolute number of circulating basophils, monocytes, lymphocytes, and PMNs, and whole blood FCER1A, CPA3 and HDC gene expression at two time points: during the anaphylactic episode and in convalescent samples collected approximately 3 months later. We then investigated the in vitro chemotactic activity of chemokines induced during anaphylaxis for the in vitro migration of the corresponding cells. Results: Only CCL2 chemokine levels were signifcantly increased in anaphylaxis samples (median 514 pg/ml) compared to convalescent samples (284 pg/ml, P<0.0001) and healthy subjects (279 pg/ml, P<0.0001)there was no signifcant diference in any of the other chemokines. There was a signifcant positive correlation between the rates of increase of serum CCL2 (median [range]: 106.0% [-44.7% to 557.4%]) and tryptase (133.8% [-6.6% to 893.4%]r=0.68, P<0.0001) and between the acute concentration of serum CCL2 and the acute concentration of serum tryptase (r=0.77, P<0.0001). The number of circulating basophils, but not other blood cells, signifcantly decreased during anaphylaxis (median 5.0 vs. 19.1 cells/[micro]l in convalescent samplesP<0.0001)a decrease in whole-blood gene expression of basophil markers (P</=0.0018) confirmed these changes. Anaphylactic serum enhances the in vitro migration of basophils via CCL2-dependent chemotactic activityin contrast, no CCL2-dependent chemotactic activity was observed for convalescent samples. Conclusions: Our findings imply an important and specifc role for CCL2-mediated chemotactic activity in the pathophysiology of human anaphylaxis

Integrative transcriptomic analysis in human and mouse model of anaphylaxis identifies gene signatures associated with cell movement, migration and neuroinflammatory signalling

BackgroundAnaphylaxis is an acute life-threatening allergic reaction and a concern at a global level; therefore, further progress in understanding the underlying mechanisms and more effective strategies for diagnosis, prevention and management are needed.ObjectiveWe sought to identify the global architecture of blood transcriptomic features of anaphylaxis by integrating expression data from human patients and mouse model of anaphylaxis.MethodsBulk RNA-sequencings of peripheral whole blood were performed in: i) 14 emergency department (ED) patients with acute anaphylaxis, predominantly to Hymenoptera venom, ii) 11 patients with peanut allergy undergoing double-blind, placebo-controlled food challenge (DBPCFC) to peanut, iii) murine model of IgE-mediated anaphylaxis. Integrative characterisation of differential gene expression, immune cell-type-specific gene expression profiles, and functional and pathway analysis was undertaken.Results1023 genes were commonly and significantly dysregulated during anaphylaxis in ED and DBPCFC patients; of those genes, 29 were also dysregulated in the mouse model. Cell-type-specific gene expression profiles showed a rapid downregulation of blood basophil and upregulation of neutrophil signature in ED and DBPCFC patients and the mouse model, but no consistent and/or significant differences were found for other blood cells. Functional and pathway analysis demonstrated that human and mouse blood transcriptomic signatures of anaphylaxis follow trajectories of upregulation of cell movement, migration and neuroinflammatory signalling, and downregulation of lipid activating nuclear receptors signalling.ConclusionOur study highlights the matched and extensive blood transcriptomic changes and suggests the involvement of discrete cellular components and upregulation of migration and neuroinflammatory pathways during anaphylaxis

POBOLJŠANJE ZBRINJAVANJA BOLESNIKA S RIJETKIM BOLESTIMA U HRVATSKOJ - TEMELJENO NA PROTOKOLU ZA HEREDITARNI ANGIOEDEM

Rare diseases affect up to 6%-8% of the population and pose a challenge to healthcare professionals and healthcare system. Hereditary angioedema is a rare life-threatening, debilitating disease characterized by recurrent edema attacks. It is essential to establish the diagnosis as quickly as possible. The Healthcare Network was created in o der to prevent mistreatment of hereditary angioedema. This study evaluated the usefulness of the Hospital Information System and the Healthcare Network in urgent management of hereditary angioedema. The Instructions for Hereditary Angioedema that contain information on the diagnosis, instructions for treatment, preventive measures prior to a dental procedure, endoscopy or surgery under general anesthesia, available regimens and storage location in the Hospital, specialist’s telephone number in emergency, and name of the family member to be contacted, were implemented in the Hospital Information System, Šibenik General Hospital as a protocol for urgent management. Data on the treatment before and after implementation of the Instructions for Hereditary Angioedema were compared. Comparing medical decisions before and after implementing the Instructions for Hereditary Angioedema in the Hospital Information System revealed that following implementation of the Instructions, correct therapy was administered more often (p=0.006, Fisher exact test) and shortterm prophylaxis applied more often before medical procedures (p=0.011, Fisher exact test). Healthcare Network raised the physician awareness of this disabling and potentially fatal disease, led to accurate diagnosis and timely treatment, enabled short hospital stay, prompt recovery, and reduced absenteeism from work due to hereditary angioedema. With specific modifications, it could also be extrapolated to other rare diseases.Rijetke bolesti su izazov zdravstvenim stručnjacima i sustavima. Smatra se da 6-8 % populacije boluje od rijetkih bolesti. Nasljedni angioedem (HAE) jedna je od rijetkih bolesti, karakterizirana ponavljajućim napadima oteklina (edema) različitih dijelova tijela te se vrlo često ne dijagnosticira pravodobno, a težina ponavljajućih napadaja se pogoršava i za život je opasno stanje. Bitno je posumnjati na rijetku bolest i postaviti dijagnozu što je brže moguće. Ova je probna studija procijenila korisnost implementacije podataka o bolesnicima s HAE u Bolnički informacijski sustav (BIS) Opće bolnice Šibenskokninske županije kao i primjenu protokola za hitno zbrinjavanje i formiranje mreže zdravstvene zaštite (MZZ). Upute u BIS-u za HAE sadrže podatke o dijagnozi HAE-a, upute za liječenje, preventivne mjere prije stomatološkog, endoskopskog ili kirurškog postupka (pod lokalnom/općom anestezijom); popis raspoloživih lijekova u hitnoći i mjesto njihove pohrane u bolnici; telefonski broj specijalista u hitnoći te ime člana obitelji (kontakt osoba). Podatci o liječenju prije primjene uputa za HAE uspoređeni su s podatcima prikupljenima u razdoblju nakon provedbe implementacije novog sustava. Uspoređujući medicinske odluke prije i nakon implementacije uputa za HAE u BIS-u, pravilna je terapija bila češće primijenjena (p =0,006, Fisherov test), kao i kratkotrajna profi laksa prije medicinskih postupaka (p = 0,011, Fisherov test). Uspostava MZZ pospješuje informiranost medicinskog osoblja, pravilan odabir liječenja i zbrinjavanje bolesnika s HAE. Omogućuje brži oporavak, kraći boravak u bolnici i smanjenje radne odsutnosti bolesnika s HAE. Temeljem ove studije otvara se mogućnost primjene MZZ i za druge rijetke bolesti

Escherichia coli Bacteriocins: Antimicrobial Efficacy and Prevalence among Isolates from Patients with Bacteraemia

Bacteriocins are antimicrobial peptides generally active against bacteria closely related to the producer. Escherichia coli produces two types of bacteriocins, colicins and microcins. The in vitro efficacy of isolated colicins E1, E6, E7, K and M, was assessed against Escherichia coli strains from patients with bacteraemia of urinary tract origin. Colicin E7 was most effective, as only 13% of the tested strains were resistant. On the other hand, 32%, 33%, 43% and 53% of the tested strains exhibited resistance to colicins E6, K, M and E1. Moreover, the inhibitory activity of individual colicins E1, E6, E7, K and M and combinations of colicins K, M, E7 and E1, E6, E7, K, M were followed in liquid broth for 24 hours. Resistance against individual colicins developed after 9 hours of treatment. On the contrary, resistance development against the combined action of 5 colicins was not observed. One hundred and five E. coli strains from patients with bacteraemia were screened by PCR for the presence of 5 colicins and 7 microcins. Sixty-six percent of the strains encoded at least one bacteriocin, 43% one or more colicins, and 54% one or more microcins. Microcins were found to co-occur with toxins, siderophores, adhesins and with the Toll/Interleukin-1 receptor domain-containing protein involved in suppression of innate immunity, and were significantly more prevalent among strains from non-immunocompromised patients. In addition, microcins were highly prevalent among non-multidrug-resistant strains compared to multidrug-resistant strains. Our results indicate that microcins contribute to virulence of E. coli instigating bacteraemia of urinary tract origin

T2-high asthma, classified by sputum mRNA expression of IL4, IL5, and IL13, is characterized by eosinophilia and severe phenotype

Asthma is a common chronic disease, with different underlying inflammatory mechanisms. Identification of asthma endotypes, which reflect a variable response to different treatments, is important for more precise asthma management. T2 asthma is characterized by airway inflammation driven by T2 cytokines including interleukins IL-4, IL-5, and IL-13. This study aimed to determine whether induced sputum samples can be used for gene expression profiling of T2-high asthma classified by IL4, IL5, and IL13 expression. Induced sputum samples were obtained from 44 subjects, among them 36 asthmatic patients and eight controls, and mRNA expression levels of IL4, IL5, and IL13 were quantified by RT-qPCR. Overall, gene expression levels of IL4, IL5, and IL13 were significantly increased in asthmatic patients\u27 samples compared to controls and there was a high positive correlation between expressions of all three genes. T2 gene mean was calculated by combining the expression levels of all three genes (IL4, IL5, and IL13) and according to T2 gene mean expression in controls, we set a T2-high/T2-low cutoff value. Twenty-four (67%) asthmatic patients had T2-high endotype and those patients had significantly higher eosinophil blood and sputum counts. Furthermore, T2-high endotype was characterized as a more severe, difficult-to-treat asthma, and often uncontrolled despite the use of inhaled and/or oral corticosteroids. Therefore, the majority of those patients (15 [63%] of 24) needed adjunct biological therapy to control their asthma symptoms/exacerbations. In conclusion, we found that interleukins IL4, IL5, and IL13 transcripts could be effectively detected in sputum from asthmatic patients. Implementation of T2 gene mean can be used as sputum molecular biomarker to categorize patients into T2-high endotype, characterized by eosinophilia and severe, difficult-to-treat asthma, and often with a need for biological treatment

Utility of telomerase gene mutation testing in patients with idiopathic pulmonary fibrosis in routine practice

Recent studies have suggested that causative variants in telomerase complex genes (TCGs) are present in around 10% of individuals with idiopathic pulmonary fibrosis (IPF) regardless of family history of the disease. However, the studies used a case-control rare variant enrichment study design which is not directly translatable to routine practice. To validate the prevalence results and to establish the individual level, routine clinical practice, and utility of those results we performed next generation sequencing of TCGs on a cohort of well-characterized consecutive individuals with IPF (diagnosis established according to ATS/ERS/JRS/ALAT guidelines). Of 27 IPF patients, three had a family history of idiopathic interstitial pneumonia (familial IPF) and 24 did not (sporadic IPF). Pathogenic/likely-pathogenic variants (according to American College of Medical Genetics criteria) in TCG were found in three individuals (11.1%) of the whole cohortspecifically, they were present in 2 out of 24 (8.3%) of the sporadic and in 1 out of 3 (33.3%) of the patients with familial IPF. Our results, which were established on an individual-patient level study design and in routine clinical practice (as opposed to the case-control study design), are roughly in line with the around 10% prevalence of causative TCG variants in patients with IPF

Damage function for poly(vinyl chloride) in heritage collections

Yellowing of plastic objects as a consequence of chemical degradation is a common heritage conservation challenge. In the case of poly(vinyl chloride) elimination of hydrogen chloride leads to the formation of polyene sequences that act as chromophores. The objective of this work was to quantitatively evaluate the rate of degradation observed as yellowing, as relevant to room conditions during long-term storage of heritage collections. Degradation was quantified as increase in the b* colour coordinate during accelerated degradation at 50 and 70 °C as a function of temperature, relative humidity, plasticizer content, and polymer molecular weight. The significance of each variable was investigated with multiple linear regression. Lower temperature, lower relative humidity, higher polymer molecular weight and higher plasticizer content were associated with lower degradation rates. The activation energy of 86 kJ/mol was calculated. The concept of ‘1- °C-equivalent’ is introduced to enable variable prioritisation from a heritage management aspect. The resulting model can be used to shape environmental management guidelines and identify the most vulnerable objects in heritage collections

Important and specific role for basophils in acute allergic reactions

IgE-mediated allergic reactions involve the activation of effector cells, predominantly through the high-affinity IgE receptor (FceRI) on mast cells and basophils. Although the mast cell is considered the major effector cell during acute allergic reactions, more recent studies indicate a potentially important and specific role for basophils and their migration which occurs rapidly upon allergen challenge in humans undergoing anaphylaxis. We review the evidence for a role of basophils in contributing to clinical symptoms of anaphylaxis, and discuss the possibility that basophil trafficking during anaphylaxis might be a pathogenic (to target organs) or protective (preventing degranulation in circulation) response. Finally, we examine the potential role of basophils in asthma exacerbations. Understanding the factors that regulate basophil trafficking and activation might lead to new diagnostic and therapeutic strategies in anaphylaxis and asthma