bluuebunny/crossref_metadata_2025 huggingface dataset:
Pankolitis Akibat Kolitis Ulseratif
Authors: Amatullah Afifah , Miro Saptino
Published: July 2021

Introduction: Ulcerative colitis is a chronic disease affecting large intestine, characterized by rectal bleeding, diarrhea, abdominal pain, and could be accompanied by extraintestinal manifestation such as fever, anemia, and weight loss. Ulcerative colitis is causing an inflammation even ulceration of the rectum which can extend to proximal colon. The incidence of this disease is increasing worldwide and its frequency is variable depend on age, race and geographic area. Thorough history and examination are needed in diagnosing ulcerative colitis, since its symptoms are similar to many other gastrointestinal diseases. Definitive diagnosis of ulcerative colitis can be obtained by colonoscopy and histopathology. Case Report: We reported a case of 53 years old man who suffered from diarrhea and weight loss for one month before admission. There were also blood and mucus in his stool. The result of colonoscopy was hyperemia and ulcers along the rectum to the caecum. Histopathology examination showed pieces of intestinal mucosal tissue covered by columnar epithelium, with goblet cell and round-oval core. The lamina propria is densely packed with lymphocyte cells, plasma cells and PMN leukocytes. This result represents chronic colitis with acute exacerbation. Conclusion: Patient was treated with mesalazine 3x1000 mg per oral, methyl prednisolone 3x16 mg per oral, and low fiber diet to achieve remission.
Salinity Stress in Wheat (Triticum aestivum L.) in the Changing Climate: Adaptation and Management Strategies
Authors: EL Sabagh Ayman , Islam Mohammad Sohidul , Skalicky Milan , Ali Raza Muhammad , Singh Kulvir , Anwar Hossain Mohammad , Hossain Akbar , Mahboob Wajid , Iqbal Muhammad Aamir , Ratnasekera Disna , Singhal Rajesh Kumar , Ahmed Sharif , Kumari Arpna , Wasaya Allah , Sytar Oksana , Brestic Marian , ÇIG Fatih , Erman Murat , Habib Ur Rahman Muhammad , Ullah Najeeb , Arshad Adnan
Published: July 2021

Wheat constitutes pivotal position for ensuring food and nutritional security; however, rapidly rising soil and water salinity pose a serious threat to its production globally. Salinity stress negatively affects the growth and development of wheat leading to diminished grain yield and quality. Wheat plants utilize a range of physiological biochemical and molecular mechanisms to adapt under salinity stress at the cell, tissue as well as whole plant levels to optimize the growth, and yield by off-setting the adverse effects of saline environment. Recently, various adaptation and management strategies have been developed to reduce the deleterious effects of salinity stress to maximize the production and nutritional quality of wheat. This review emphasizes and synthesizes the deleterious effects of salinity stress on wheat yield and quality along with highlighting the adaptation and mitigation strategies for sustainable wheat production to ensure food security of skyrocketing population under changing climate.
Artificial Intelligence Enhances Studies on Inflammatory Bowel Disease
Authors: Chen Guihua , Shen Jun
Published: July 2021

Inflammatory bowel disease (IBD), which includes ulcerative colitis (UC) and Crohn’s disease (CD), is an idiopathic condition related to a dysregulated immune response to commensal intestinal microflora in a genetically susceptible host. As a global disease, the morbidity of IBD reached a rate of 84.3 per 100,000 persons and reflected a continued gradual upward trajectory. The medical cost of IBD is also notably extremely high. For example, in Europe, it has €3,500 in CD and €2,000 in UC per patient per year, respectively. In addition, taking into account the work productivity loss and the reduced quality of life, the indirect costs are incalculable. In modern times, the diagnosis of IBD is still a subjective judgment based on laboratory tests and medical images. Its early diagnosis and intervention is therefore a challenging goal and also the key to control its progression. Artificial intelligence (AI)-assisted diagnosis and prognosis prediction has proven effective in many fields including gastroenterology. In this study, support vector machines were utilized to distinguish the significant features in IBD. As a result, the reliability of IBD diagnosis due to its impressive performance in classifying and addressing region problems was improved. Convolutional neural networks are advanced image processing algorithms that are currently in existence. Digestive endoscopic images can therefore be better understood by automatically detecting and classifying lesions. This study aims to summarize AI application in the area of IBD, objectively evaluate the performance of these methods, and ultimately understand the algorithm–dataset combination in the studies.
Bruton’s Tyrosine Kinase and Its Isoforms in Cancer
Authors: Wang Xianhui , Kokabee Leila , Kokabee Mostafa , Conklin Douglas S.
Published: July 2021

Bruton’s tyrosine kinase (BTK) is a soluble tyrosine kinase with central roles in the development, maturation, and signaling of B cells. BTK has been found to regulate cell proliferation, survival, and migration in various B-cell malignancies. Targeting BTK with recently developed BTK inhibitors has been approved by the Food and Drug Administration (FDA) for the treatment of several hematological malignancies and has transformed the treatment of several B-cell malignancies. The roles that BTK plays in B cells have been appreciated for some time. Recent studies have established that BTK is expressed and plays pro-tumorigenic roles in several epithelial cancers. In this review, we focus on novel isoforms of the BTK protein expressed in epithelial cancers. We review recent work on the expression, function, and signaling of these isoforms and their value as potential therapeutic targets in epithelial tumors.
Potential Impact of Diabetes and Obesity on Alveolar Type 2 (AT2)-Lipofibroblast (LIF) Interactions After COVID-19 Infection
Authors: Nouri-Keshtkar Marjan , Taghizadeh Sara , Farhadi Aisan , Ezaddoustdar Aysan , Vesali Samira , Hosseini Roya , Totonchi Mehdi , Kouhkan Azam , Chen Chengshui , Zhang Jin-San , Bellusci Saverio , Tahamtani Yaser
Published: July 2021

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a new emerging respiratory virus, caused evolving pneumonia outbreak around the world. In SARS-Cov-2 infected patients, diabetes mellitus (DM) and obesity are two metabolic diseases associated with higher severity of SARS-CoV-2 related complications, characterized by acute lung injury requiring assisted ventilation as well as fibrosis development in surviving patients. Different factors are potentially responsible for this exacerbated response to SARS-CoV-2 infection. In patients with DM, base-line increase in inflammation and oxidative stress represent preexisting risk factors for virus-induced damages. Such factors are also likely to be found in obese patients. In addition, it has been proposed that massive injury to the alveolar epithelial type 2 (AT2) cells, which express the SARS-CoV-2 receptor angiotensin-converting enzyme 2 (ACE2), leads to the activation of their stromal niches represented by the Lipofibroblasts (LIF). LIF are instrumental in maintaining the self-renewal of AT2 stem cells. LIF have been proposed to transdifferentiate into Myofibroblast (MYF) following injury to AT2 cells, thereby contributing to fibrosis. We hypothesized that LIF’s activity could be impacted by DM or obesity in an age- and gender-dependent manner, rendering them more prone to transition toward the profibrotic MYF status in the context of severe COVID-19 pneumonia. Understanding the cumulative effects of DM and/or obesity in the context of SARS-CoV-2 infection at the cellular level will be crucial for efficient therapeutic solutions.
The Sensitivity of the Marine Carbonate System to Regional Ocean Alkalinity Enhancement
Authors: Burt Daniel J. , Fröb Friederike , Ilyina Tatiana
Published: July 2021

Ocean Alkalinity Enhancement (OAE) simultaneously mitigates atmospheric concentrations of CO2 and ocean acidification; however, no previous studies have investigated the response of the non-linear marine carbonate system sensitivity to alkalinity enhancement on regional scales. We hypothesise that regional implementations of OAE can sequester more atmospheric CO2 than a global implementation. To address this, we investigate physical regimes and alkalinity sensitivity as drivers of the carbon-uptake potential response to global and different regional simulations of OAE. In this idealised ocean-only set-up, total alkalinity is enhanced at a rate of 0.25 Pmol a-1 in 75-year simulations using the Max Planck Institute Ocean Model coupled to the HAMburg Ocean Carbon Cycle model with pre-industrial atmospheric forcing. Alkalinity is enhanced globally and in eight regions: the Subpolar and Subtropical Atlantic and Pacific gyres, the Indian Ocean and the Southern Ocean. This study reveals that regional alkalinity enhancement has the capacity to exceed carbon uptake by global OAE. We find that 82–175 Pg more carbon is sequestered into the ocean when alkalinity is enhanced regionally and 156 PgC when enhanced globally, compared with the background-state. The Southern Ocean application is most efficient, sequestering 12% more carbon than the Global experiment despite OAE being applied across a surface area 40 times smaller. For the first time, we find that different carbon-uptake potentials are driven by the surface pattern of total alkalinity redistributed by physical regimes across areas of different carbon-uptake efficiencies. We also show that, while the marine carbonate system becomes less sensitive to alkalinity enhancement in all experiments globally, regional responses to enhanced alkalinity vary depending upon the background concentrations of dissolved inorganic carbon and total alkalinity. Furthermore, the Subpolar North Atlantic displays a previously unexpected alkalinity sensitivity increase in response to high total alkalinity concentrations.
Combinatorial Analysis of Phenotypic and Clinical Risk Factors Associated With Hospitalized COVID-19 Patients
Authors: Das Sayoni , Pearson Matthew , Taylor Krystyna , Bouchet Veronique , Møller Gert Lykke , Hall Taryn O. , Strivens Mark , Tzeng Kathy T. H. , Gardner Steve
Published: July 2021

Characterization of the risk factors associated with variability in the clinical outcomes of COVID-19 is important. Our previous study using genomic data identified a potential role of calcium and lipid homeostasis in severe COVID-19. This study aimed to identify similar combinations of features (disease signatures) associated with severe disease in a separate patient population with purely clinical and phenotypic data. The PrecisionLife combinatorial analytics platform was used to analyze features derived from de-identified health records in the UnitedHealth Group COVID-19 Data Suite. The platform identified and analyzed 836 disease signatures in two cohorts associated with an increased risk of COVID-19 hospitalization. Cohort 1 was formed of cases hospitalized with COVID-19 and a set of controls who developed mild symptoms. Cohort 2 included Cohort 1 individuals for whom additional laboratory test data was available. We found several disease signatures where lower levels of lipids were found co-occurring with lower levels of serum calcium and leukocytes. Many of the low lipid signatures were independent of statin use and 50% of cases with hypocalcemia signatures were reported with vitamin D deficiency. These signatures may be attributed to similar mechanisms linking calcium and lipid signaling where changes in cellular lipid levels during inflammation and infection affect calcium signaling in host cells. This study and our previous genomics analysis demonstrate that combinatorial analysis can identify disease signatures associated with the risk of developing severe COVID-19 separately from genomic or clinical data in different populations. Both studies suggest associations between calcium and lipid signaling in severe COVID-19.
Attitudes Toward Health, Healthcare, and eHealth of People With a Low Socioeconomic Status: A Community-Based Participatory Approach
Authors: Faber Jasper S. , Al-Dhahir Isra , Reijnders Thomas , Chavannes Niels H. , Evers Andrea W. M. , Kraal Jos J. , van den Berg-Emons H. J. G. , Visch Valentijn T.
Published: July 2021

Low socioeconomic status (SES) is associated with a higher prevalence of unhealthy lifestyles compared to a high SES. Health interventions that promote a healthy lifestyle, like eHealth solutions, face limited adoption in low SES groups. To improve the adoption of eHealth interventions, their alignment with the target group's attitudes is crucial. This study investigated the attitudes of people with a low SES toward health, healthcare, and eHealth. We adopted a mixed-method community-based participatory research approach with 23 members of a community center in a low SES neighborhood in the city of Rotterdam, the Netherlands. We conducted a first set of interviews and analyzed these using a grounded theory approach resulting in a group of themes. These basic themes' representative value was validated and refined by an online questionnaire involving a different sample of 43 participants from multiple community centers in the same neighborhood. We executed three focus groups to validate and contextualize the results. We identified two general attitudes based on nine profiles toward health, healthcare, and eHealth. The first general attitude, optimistically engaged, embodied approximately half our sample and involved light-heartedness toward health, loyalty toward healthcare, and eagerness to adopt eHealth. The second general attitude, doubtfully disadvantaged, represented roughly a quarter of our sample and was related to feeling encumbered toward health, feeling disadvantaged within healthcare, and hesitance toward eHealth adoption. The resulting attitudes strengthen the knowledge of the motivation and behavior of people with low SES regarding their health. Our results indicate that negative health attitudes are not as evident as often claimed. Nevertheless, intervention developers should still be mindful of differentiating life situations, motivations, healthcare needs, and eHealth expectations. Based on our findings, we recommend eHealth should fit into the person's daily life, ensure personal communication, be perceived usable and useful, adapt its communication to literacy level and life situation, allow for meaningful self-monitoring and embody self-efficacy enhancing strategies.
CRISPR/Cas9-Mediated Multi-Allelic Gene Targeting in Sugarcane Confers Herbicide Tolerance
Authors: Oz Mehmet Tufan , Altpeter Angelika , Karan Ratna , Merotto Aldo , Altpeter Fredy
Published: July 2021

Sugarcane is the source of 80% of the sugar and 26% of the bioethanol produced globally. However, its complex, highly polyploid genome (2n= 100 – 120) impedes crop improvement. Here, we report efficient and reproducible gene targeting (GT) in sugarcane, enabling precise co-editing of multiple allelesviatemplate-mediated and homology-directed repair (HDR) of DNA double strand breaks induced by the programmable nuclease CRISPR/Cas9. The evaluation of 146 independently transformed plants from five independent experiments revealed a targeted nucleotide replacement that resulted in both targeted amino acid substitutions W574L and S653I in the acetolactate synthase (ALS) in 11 lines in addition to single, targeted amino acid substitutions W574L or S653I in 25 or 18 lines, respectively. Co-editing of up to three ALS copies/alleles that confer herbicide tolerance was confirmed by Sanger sequencing of cloned long polymerase chain reaction (PCR) amplicons. This work will enable crop improvement by conversion of inferior alleles to superior alleles through targeted nucleotide substitutions.
Exact Probability Landscapes of Stochastic Phenotype Switching in Feed-Forward Loops: Phase Diagrams of Multimodality
Authors: Terebus Anna , Manuchehrfar Farid , Cao Youfang , Liang Jie
Published: July 2021

Feed-forward loops (FFLs) are among the most ubiquitously found motifs of reaction networks in nature. However, little is known about their stochastic behavior and the variety of network phenotypes they can exhibit. In this study, we provide full characterizations of the properties of stochastic multimodality of FFLs, and how switching between different network phenotypes are controlled. We have computed the exact steady-state probability landscapes of all eight types of coherent and incoherent FFLs using the finite-butter Accurate Chemical Master Equation (ACME) algorithm, and quantified the exact topological features of their high-dimensional probability landscapes using persistent homology. Through analysis of the degree of multimodality for each of a set of 10,812 probability landscapes, where each landscape resides over 105–106 microstates, we have constructed comprehensive phase diagrams of all relevant behavior of FFL multimodality over broad ranges of input and regulation intensities, as well as different regimes of promoter binding dynamics. In addition, we have quantified the topological sensitivity of the multimodality of the landscapes to regulation intensities. Our results show that with slow binding and unbinding dynamics of transcription factor to promoter, FFLs exhibit strong stochastic behavior that is very different from what would be inferred from deterministic models. In addition, input intensity play major roles in the phenotypes of FFLs: At weak input intensity, FFL exhibit monomodality, but strong input intensity may result in up to 6 stable phenotypes. Furthermore, we found that gene duplication can enlarge stable regions of specific multimodalities and enrich the phenotypic diversity of FFL networks, providing means for cells toward better adaptation to changing environment. Our results are directly applicable to analysis of behavior of FFLs in biological processes such as stem cell differentiation and for design of synthetic networks when certain phenotypic behavior is desired.
Whole-Genome Resequencing to Study Brucellosis Susceptibility in Sheep
Authors: Li Xiaolong , Wu Qingmin , Zhang Xiaoxue , Li Chong , Zhang Deyin , Li Guoze , Zhang Yukun , Zhao Yuan , Shi Zhaoguo , Wang Weimin , Li Fadi
Published: July 2021

Brucellosis is a zoonotic disease and a major public health problem. However, the genetic mechanism of brucellosis in sheep remains unclear. In this study, serum samples were collected from 6,358 sheep from the F2 population (Dorper sheep ♂ × Hu sheep ♀), and antibody levels were continuously measured at 14 days and 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10 months after administration of brucellosis vaccine. Finally, 19 brucellosis-resistant group (BRG) sheep and 22 brucellosis-susceptible group sheep (BSG) were screened for whole-genome sequencing. Using the fixation index, Fisher’s exact test, and chi-square test, a total of 205 candidate SNP sites were identified. Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis suggested that 138 candidate genes were significantly enriched in adherens junction (CTNNA3, PARD3, and PTPRM), cell adhesion molecules (NLGN1, CNTNAP2, NCAM1, and PTPRM), salivary secretion (LOC101102109, PRKG1, and ADCY2), and hippo signaling pathway (CTNNA3, YAP1, and PARD3). These findings provide valuable molecular markers for brucellosis resistance breeding in sheep and novel insights into the genetic mechanism of brucellosis resistance.
Association Analysis of Candidate Variants in Admixed Brazilian Patients With Genetic Generalized Epilepsies
Authors: Kaibara Felipe S. , de Araujo Tânia K. , Araujo Patricia A. O. R. A. , Alvim Marina K. M. , Yasuda Clarissa L. , Cendes Fernando , Lopes-Cendes Iscia , Secolin Rodrigo
Published: July 2021

Genetic generalized epilepsies (GGEs) include well-established epilepsy syndromes with generalized onset seizures: childhood absence epilepsy, juvenile myoclonic epilepsy (JME), juvenile absence epilepsy (JAE), myoclonic absence epilepsy, epilepsy with eyelid myoclonia (Jeavons syndrome), generalized tonic–clonic seizures, and generalized tonic–clonic seizures alone. Genome-wide association studies (GWASs) and exome sequencing have identified 48 single-nucleotide polymorphisms (SNPs) associated with GGE. However, these studies were mainly based on non-admixed, European, and Asian populations. Thus, it remains unclear whether these results apply to patients of other origins. This study aims to evaluate whether these previous results could be replicated in a cohort of admixed Brazilian patients with GGE. We obtained SNP-array data from 87 patients with GGE, compared with 340 controls from the BIPMed public dataset. We could directly access genotypes of 17 candidate SNPs, available in the SNP array, and the remaining 31 SNPs were imputed using the BEAGLE v5.1 software. We performed an association test by logistic regression analysis, including the first five principal components as covariates. Furthermore, to expand the analysis of the candidate regions, we also interrogated 14,047 SNPs that flank the candidate SNPs (1 Mb). The statistical power was evaluated in terms of odds ratio and minor allele frequency (MAF) by the genpwr package. Differences in SNP frequencies between Brazilian and Europeans, sub-Saharan African, and Native Americans were evaluated by a two-proportion Z-test. We identified nine flanking SNPs, located on eight candidate regions, which presented association signals that passed the Bonferroni correction (rs12726617; rs9428842; rs1915992; rs1464634; rs6459526; rs2510087; rs9551042; rs9888879; and rs8133217; p-values <3.55e–06). In addition, the two-proportion Z-test indicates that the lack of association of the remaining candidate SNPs could be due to different genomic backgrounds observed in admixed Brazilians. This is the first time that candidate SNPs for GGE are analyzed in an admixed Brazilian population, and we could successfully replicate the association signals in eight candidate regions. In addition, our results provide new insights on how we can account for population structure to improve risk stratification estimation in admixed individuals.
Causal Associations of Urate With Cardiovascular Risk Factors: Two-Sample Mendelian Randomization
Authors: Lukkunaprasit Thitiya , Rattanasiri Sasivimol , Ongphiphadhanakul Boonsong , McKay Gareth J. , Attia John , Thakkinstian Ammarin
Published: July 2021

Background

Mendelian Randomization (MR) studies show conflicting causal associations of genetically predicted serum urate with cardiovascular risk factors (i.e., hypertension, diabetes, lipid profile, and kidney function). This study aimed to robustly investigate a causal relationship between urate and cardiovascular risk factors considering single nucleotide polymorphisms (SNPs) as instrumental variables using two-sample MR and various sensitivity analyses.

Methods

Data on SNP-urate associations were taken from the Global Urate Genetics Consortium and data on SNP-cardiovascular risk factor associations were taken from various consortia/UK Biobank. SNPs were selected by statistically and biologically driven approaches as instrumental variables. Various sensitivity analyses were performed using different MR methods including inverse variance weighted, MR-Egger, weighted median/mode, MR-PRESSO, and the contamination mixture method.

Results

The statistically driven approach showed significant causal effects of urate on HDL-C and triglycerides using four of the six MR methods, i.e., every 1 mg/dl increase in genetically predicted urate was associated with 0.047 to 0.103 SD decrease in HDL-C and 0.034 to 0.207 SD increase in triglycerides. The biologically driven approach to selection of SNPs from ABCG2, SLC2A9, SLC17A1, SLC22A11, and SLC22A12 showed consistent causal effects of urate on HDL-C from all methods with 0.038 to 0.057 SD decrease in HDL-C per 1 mg/dl increase of urate, and no evidence of horizontal pleiotropy was detected.

Conclusion

Our study suggests a significant and robust causal effect of genetically predicted urate on HDL-C. This finding may explain a small proportion (7%) of the association between increased urate and cardiovascular disease but points to urate being a novel cardiac risk factor.
CYP3A4∗22 Genotyping in Clinical Practice: Ready for Implementation?
Authors: Mulder Tessa A. M. , van Eerden Ruben A. G. , de With Mirjam , Elens Laure , Hesselink Dennis A. , Matic Maja , Bins Sander , Mathijssen Ron H. J. , van Schaik Ron H. N.
Published: July 2021

Cytochrome P450 3A4 (CYP3A4) is the most important drug metabolizing enzyme in the liver, responsible for the oxidative metabolism of ∼50% of clinically prescribed drugs. Therefore, genetic variation inCYP3A4could potentially affect the pharmacokinetics, toxicity and clinical outcome of drug treatment. Thus far, pharmacogenetics for CYP3A4 has not received much attention. However, the recent discovery of the intron 6 single-nucleotide polymorphism (SNP) rs35599367C > T, encoding theCYP3A4∗22allele, led to several studies into the pharmacogenetic effect ofCYP3A4∗22on different drugs. This allele has a relatively minor allele frequency of 3-5% and an effect on CYP3A4 enzymatic activity. Thus far, no review summarizing the data published on several drugs is available yet. This article therefore addresses the current knowledge onCYP3A4∗22. This information may help in deciding if, and for which drugs,CYP3A4∗22genotype-based dosing could be helpful in improving drug therapy.CYP3A4∗22was shown to significantly influence the pharmacokinetics of several drugs, with currently being most thoroughly investigated tacrolimus, cyclosporine, and statins. Additional studies, focusing on toxicity and clinical outcome, are warranted to demonstrate clinical utility ofCYP3A4∗22genotype-based dosing.
Early Pup Removal Leads to Social Dysfunction and Dopamine Deficit in Late Postpartum Rats: Prevention by Social Support
Authors: Rincón-Cortés Millie , Grace Anthony A.
Published: July 2021

Offspring interaction is among the most highly motivated behaviors in maternal mammals and is mediated by mesolimbic dopamine (DA) system activation. Disruption or loss of significant social relationships is among the strongest individual predictors of affective dysregulation and depression onset in humans. However, little is known regarding the effects of disrupted mother–infant attachment (pup removal) in rat dams. Here, we tested the effects of permanent pup removal in rat dams, which were assigned to one of three groups on postpartum day (PD) 1: pups; pups removed, single-housed; or pups removed, co-housed with another dam who also had pups removed; and underwent a behavioral test battery during PD 21–23.In vivoelectrophysiological recordings of ventral tegmental area (VTA) DA neurons were performed on PD 22 and 23 in a subset of animals. Pup removal did not impact sucrose consumption or anxiety-like behavior, but increased passive forced swim test (FST) coping responses. Pup-removal effects on social behavior and VTA activity were sensitive to social buffering: only single-housed dams exhibited reduced social motivation and decreased numbers of active DA neurons. Dams that had pups removed and were co-housed did not exhibit changes in social behavior or VTA function. Moreover, no changes in social behavior, FST coping, or VTA activity were found in socially isolated adult virgin females, indicating that effects observed in dams are specific to pup loss. This study showed that deprivation of species-expected social relationships (pups) during the postpartum precipitates an enduring negative affect state (enhanced passive coping, blunted social motivation) and attenuated VTA DA function in the dam, and that a subset of these effects is partially ameliorated through social buffering.
The Role of Regulatory T Cells in Epicutaneous Immunotherapy for Food Allergy
Authors: Liu Guirong , Liu Manman , Wang Junjuan , Mou Yao , Che Huilian
Published: July 2021

In recent decades, a rapid increase in the prevalence of food allergies has led to extensive research on novel treatment strategies and their mechanisms. Mouse models have provided preliminary insights into the mechanism of epicutaneous immunotherapy (EPIT)-induced immune tolerance. In EPIT, antigen applied on the skin surface can be captured, processed, and presented in the lymph nodes (LNs) by Antigen-presenting cells (APCs). In the LNs, induction of regulatory T cells (Treg cells) requires both direct contact during antigen presentation and indirect mechanisms such as cytokines. Foxp3+CD62L+ Treg cells can exhibit the characteristics of hypomethylation of Foxp3 TSDR and Foxp3-LAP+ Treg cells, which increase the expression of surface tissue-specific homing molecules to exert further sustained systemic immune tolerance. Studies have shown that EPIT is a potential treatment for food allergies and can effectively induce immune tolerance, but its mechanism needs further exploration. Here, we review Treg cells’ role in immune tolerance induced by EPIT and provide a theoretical basis for future research directions, such as the mechanism of EPIT and the development of more effective EPIT treatments.
A Stable Cell Line Expressing Clustered AChR: A Novel Cell-Based Assay for Anti-AChR Antibody Detection in Myasthenia Gravis
Authors: Cai Yu , Han Lu , Zhu Desheng , Peng Jing , Li Jianping , Ding Jie , Luo Jiaying , Hong Ronghua , Wang Kan , Wan Wenbin , Xie Chong , Zhou Xiajun , Zhang Ying , Hao Yong , Guan Yangtai
Published: July 2021

Cell-based assays (CBAs) and radioimmunoprecipitation assay (RIPA) are the most sensitive methods for identifying anti-acetylcholine receptor (AChR) antibody in myasthenia gravis (MG). But CBAs are limited in clinical practice by transient transfection. We established a stable cell line (KL525) expressing clustered AChR by infecting HEK 293T cells with dual lentiviral vectors expressing the genes encoding the human AChR α1, β1, δ, ϵ and the clustering protein rapsyn. We verified the stable expression of human clustered AChR by immunofluorescence, immunoblotting, and real-time PCR. Fluorescence-activated cell sorting (FACS) was used to detect anti-AChR antibodies in 103 MG patients and 58 healthy individuals. The positive results of MG patients reported by the KL525 was 80.6% (83/103), 29.1% higher than the 51.4% (53/103) of RIPA. 58 healthy individuals tested by both the KL525 CBA and RIPA were all negative. In summary, the stable expression of clustered AChR in our cell line makes it highly sensitive and advantageous for broad clinical application in CBAs.
Clickable Vitamins as a New Tool to Track Vitamin A and Retinoic Acid in Immune Cells
Authors: Bos Amelie V. , Erkelens Martje N. , Koenders Sebastiaan T.A. , van der Stelt Mario , van Egmond Marjolein , Mebius Reina E.
Published: July 2021

The vitamin A derivative, retinoid acid (RA) is key player in guiding adaptive mucosal immune responses. However, data on the uptake and metabolism of vitamin A within human immune cells has remained largely elusive because retinoids are small, lipophilic molecules which are difficult to detect. To overcome this problem and to be able to study the effect of vitamin A metabolism in human immune cell subsets, we have synthesized novel bio-orthogonal retinoid-based probes (clickable probes), which are structurally and functionally indistinguishable from vitamin A. The probes contain a functional group (an alkyne) to conjugate to a fluorogenic dye to monitor retinoid molecules in real-time in immune cells. We demonstrate, by using flow cytometry and microscopy, that multiple immune cells have the capacity to internalize retinoids to varying degrees, including human monocyte-derived dendritic cells (DCs) and naïve B lymphocytes. We observed that naïve B cells lack the enzymatic machinery to produce RA, but use exogenous retinoic acid to enhance CD38 expression. Furthermore, we showed that human DCs metabolize retinal into retinoic acid, which in co-culture with naïve B cells led to of the induction of CD38 expression. These data demonstrate that in humans, DCs can serve as an exogenous source of RA for naïve B cells. Taken together, through the use of clickable vitamins our data provide valuable insight in the mechanism of vitamin A metabolism and its importance for human adaptive immunity.
Development of a Potential Yeast-Based Vaccine Platform for Theileria parva Infection in Cattle
Authors: Goh Shan , Kolakowski Jeannine , Holder Angela , Pfuhl Mark , Ngugi Daniel , Ballingall Keith , Tombacz Kata , Werling Dirk
Published: July 2021

East Coast Fever (ECF), caused by the tick-borne apicomplexan parasiteTheileria parva, remains one of the most important livestock diseases in sub-Saharan Africa with more than 1 million cattle dying from infection every year. Disease prevention relies on the so-called “Infection and Treatment Method” (ITM), which is costly, complex, laborious, difficult to standardise on a commercial scale and results in a parasite strain-specific, MHC class I-restricted cytotoxic T cell response. We therefore attempted to develop a safe, affordable, stable, orally applicable and potent subunit vaccine for ECF using five differentT. parvaschizont antigens (Tp1, Tp2, Tp9, Tp10 and N36) andSaccharomyces cerevisiaeas an expression platform. Full-length Tp2 and Tp9 as well as fragments of Tp1 were successfully expressed on the surface ofS. cerevisiae.In vitroanalyses highlighted that recombinant yeast expressing Tp2 can elicit IFNγ responses using PBMCs from ITM-immunized calves, while Tp2 and Tp9 induced IFNγ responses from enriched bovine CD8+T cells. A subsequentin vivostudy showed that oral administration of heat-inactivated, freeze-dried yeast stably expressing Tp2 increased total murine serum IgG over time, but more importantly, induced Tp2-specific serum IgG antibodies in individual mice compared to the control group. While these results will require subsequent experiments to verify induction of protection in neonatal calves, our data indicates that oral application of yeast expressing Theileria antigens could provide an affordable and easy vaccination platform for sub-Saharan Africa. Evaluation of antigen-specific cellular immune responses, especially cytotoxic CD8+T cell immunity in cattle will further contribute to the development of a yeast-based vaccine for ECF.
Comparative Analysis of the Measles Antibody Levels in Healthy Medical Personnel of Maternity Ward and Women in Labor
Authors: Kostinov Mikhail Petrovich , Zhuravlev Pavel Ivanovich , Gladkova Lylia Solomonovna , Mashilov Kirill Vadimovich , Polishchuk Valentina Borisovna , Shmitko Anna Dmitrievna , Zorina Veronika Nikolaevna , Blagovidov Dmitriy Alexeyevich , Pahomov Dmitriy Vladimirovich , Vlasenko Anna Egorovna , Ryzhov Alexey Anatolevich , Khromova Ekaterina Alexandrovna
Published: July 2021

It has been proven that post-vaccination immunity to measles virus after two doses of vaccine is not able to persistently protect against infection throughout life. The goal of this research was to determine the immune layer to the measles virus among women in labor and maternity ward personnel in the same medical institution. The levels of IgG antibodies to measles virus in the umbilical cord blood of 594 women in labor and 88 workers of the maternity ward were studied by ELISA. It was revealed that 22.7% of umbilical cord blood serum samples from parturient women and 21.4% of blood serum samples from maternity ward personnel were seronegative (<0.18 IU/ml). Levels of IgG antibodies to measles virus in low values (<1.0 IU/ml) were detected in 67% of blood serum samples among women in labor and 68.9% among employees of the maternity ward. Among women in labor, women under 35 years of age are at the highest risk of contracting measles; the proportion of women with low levels of protective antibodies in this age group was almost 70%, and the proportion of women without protective levels of antibodies was 23%. Compared with the age group 36–43, the age of women in labor under 35 was associated with a higher chance of not having immune protection against infection with measles virus OR [95% CI] = 2.2 [1.1–4.5] (p = 0.02) or had a low level of protection OR [95% CI] = 1.9 [1.2–3.0] (p = 0.001). It was also found that among women over 35 years of age, the proportion of persons with a high level of antibodies in women in labor was statistically significantly higher than among members of the maternity ward staff (13 and 0%, respectively, p = 0.007). Thus, maternity ward employees and women in labor constitute a risk group for measles due to the presence of a high proportion of seronegative persons among women of childbearing age (both maternity ward employees and women in labor). These conditions create the need to revise current approaches to present vaccination procedures, especially in the current epidemiological situation with COVID-19.
Transgenic Expression of a Mutant Ribonuclease Regnase-1 in T Cells Disturbs T Cell Development and Functions
Authors: Kong Gangcheng , Dou Yaling , Xiao Xiang , Wang Yixuan , Ming Yingzi , Li Xian C.
Published: July 2021

Regnase-1 is an RNA-binding protein with ribonuclease activities, and once induced it controls diverse immune responses by degrading mRNAs that encode inflammatory cytokines and costimulatory molecules, thus exerting potent anti-inflammatory functions. However, Regnase-1 is extremely sensitive to degradation by proteases and therefore short-lived. Here, we constructed a mutant Regnase-1 that is resistant to degradation and expressed this mutantin vivoas a transgene specifically in T cells. We found that the mutant Regnase-1 transgenic mice exhibited profound lymphopenia in the periphery despite grossly normal spleen and lymph nodes, and spontaneously accepted skin allografts without any treatment. Mechanistic studies showed that in the transgenic mice thymic T cell development was disrupted, such that most of the developing thymocytes were arrested at the double positive stage, with few mature CD4+and CD8+T cells in the thymus and periphery. Our findings suggest that interfering with the dynamic Regnase-1 expression in T cells disrupts T cell development and functions and further studies are warranted to uncover the mechanisms involved.
Altered Functions of Neutrophils in Two Chinese Patients With Severe Congenital Neutropenia Type 4 Caused by G6PC3 Mutations
Authors: Dai Rongxin , Lv Ge , Li Wenyan , Tang Wenjing , Chen Junjie , Liu Qiao , Yang Lu , Zhang Min , Tian Zhirui , Zhou Lina , Yan Xin , Wang Yating , Ding Yuan , An Yunfei , Zhang Zhiyong , Tang Xuemei , Zhao Xiaodong
Published: July 2021

Background

SCN4 is an autosomal recessive disease caused by mutations in the G6PC3 gene. The clinical, molecular, and immunological features; function of neutrophils; and prognosis of patients with SCN4 have not been fully elucidated.

Methods

Two Chinese pediatric patients with G6PC3 mutations were enrolled in this study. Clinical data, genetic and immunologic characteristics, and neutrophil function were evaluated in patients and controls before and after granulocyte colony-stimulating factor (G-CSF) treatment.

Results

Both patients had histories of pneumonia, inguinal hernia, cryptorchidism, and recurrent oral ulcers. Patient 1 also had asthma and otitis media, and patient 2 presented with prominent ectatic superficial veins and inflammatory bowel disease. DNA sequencing demonstrated that both patients harbored heterozygous G6PC3 gene mutations. Spontaneous and FAS-induced neutrophil apoptosis were significantly increased in patients, and improved only slightly after G-CSF treatment, while neutrophil respiratory burst and neutrophil extracellular traps production remained impaired in patients after G-CSF treatment.

Conclusion

G-CSF treatment is insufficient for patients with SCN4 patients, who remain at risk of infection. Where possible, regular G-CSF treatment, long-term prevention of infection, are the optimal methods for cure of SCN4 patients. It is important to monitor closely for signs of leukemia in SCN4 patients. Once leukemia occurs in SCN4 patients, hematopoietic stem cell transplantation is the most important choice of treatment.
Desiccation Induced Conjunctival Monocyte Recruitment and Activation - Implications for Keratoconjunctivitis
Authors: Alam Jehan , de Paiva Cintia S. , Pflugfelder Stephen C.
Published: July 2021

Background

Lacrimal gland secretory dysfunction in Sjögren syndrome (SS) causes ocular surface desiccation that is associated with increased cytokine expression and number of antigen-presenting cells (APCs) in the conjunctiva. This study evaluated the hypothesis that desiccating stress (DS) alters the percentage and gene expression of myeloid cell populations in the conjunctiva.

Methods

DS was induced by pharmacologic suppression of tear secretion and exposure to drafty low humidity environment. Bone marrow chimeras and adoptive transfer of CD45.1+ bone marrow cells to CD45.2+ C-C chemokine receptor 2 knockout (CCR2-/-) mice were used to track DS-induced myeloid cell recruitment to the conjunctiva. Flow cytometry evaluated myeloid cell populations in conjunctivae obtained from non-stressed eyes and those exposed to DS for 5 days. CD11b+ myeloid lineage cells were gated on monocyte (Ly6C), macrophage (CD64, MHCII) and DC (CD11c, MHCII) lineage markers. NanoString immune arrays were performed on sorted MHCII+ and MHCII- monocyte/macrophage cell populations.

Results

DS significantly increased the recruitment of adoptively transferred MHCII positive and negative myeloid cells to the conjunctiva in a CCR2 dependent fashion. The percentage of resident conjunctival monocytes (Ly6C+CD64-) significantly decreased while CD64+MHCII+ macrophages increased over 5 days of DS (P<0.05 for both). Comparison of gene expression between the MHCII- monocyte and MHCII+ populations in non-stressed conjunctiva revealed a ≥ 2 log2 fold increase in 95 genes and decrease in 46 genes. Upregulated genes are associated with antigen presentation, cytokine/chemokine, M1 macrophage and NLRP3 inflammasome pathways. DS increased innate inflammatory genes in monocytes and MHCII+ cells and increased M1 macrophage (Trem1, Ido1, Il12b, Stat5b) and decreased homeostasis (Mertk) and M2 macrophage (Arg1) genes in MHCII+ cells.

Conclusions

There are myeloid cell populations in the conjunctiva with distinct phenotype and gene expression patterns. DS recruits myeloid cells from the blood and significantly changes their phenotype in the conjunctiva. DS also alters expression of an array of innate inflammatory genes. Immature monocytes in the unstressed conjunctiva appear to cascade to MHCII+ macrophages during DS, suggesting that DS promotes maturation of monocytes to antigen presenting cells with increased expression of inflammatory genes that may contribute to the pathogenesis of SS keratoconjunctivitis sicca.
IL-10-Dependent Amelioration of Chronic Inflammatory Disease by Microdose Subcutaneous Delivery of a Prototypic Immunoregulatory Small Molecule
Authors: Tabares-Guevara Jorge H. , Jaramillo Julio C. , Ospina-Quintero Laura , Piedrahíta-Ochoa Christian A. , García-Valencia Natalia , Bautista-Erazo David E. , Caro-Gómez Erika , Covián Camila , Retamal-Díaz Angello , Duarte Luisa F. , González Pablo A. , Bueno Susan M. , Riedel Claudia A. , Kalergis Alexis M. , Ramírez-Pineda José R.
Published: July 2021

One of the interventional strategies to reestablish the immune effector/regulatory balance, that is typically altered in chronic inflammatory diseases (CID), is the reinforcement of endogenous immunomodulatory pathways as the one triggered by interleukin (IL)-10. In a recent work, we demonstrated that the subcutaneous (sc) administration of an IL-10/Treg-inducing small molecule-based formulation, using a repetitive microdose (REMID) treatment strategy to preferentially direct the effects to the regional immune system, delays the progression of atherosclerosis. Here we investigated whether the same approach using other IL-10-inducing small molecule, such as the safe, inexpensive, and widely available polyphenol curcumin, could induce a similar protective effect in two different CID models. We found that, in apolipoprotein E deficient mice, sc treatment with curcumin following the REMID strategy induced atheroprotection that was not consequence of its direct systemic lipid-modifying or antioxidant activity, but instead paralleled immunomodulatory effects, such as reduced proatherogenic IFNγ/TNFα-producing cells and increased atheroprotective FOXP3+Tregs and IL-10-producing dendritic and B cells. Remarkably, when a similar strategy was used in the neuroinflammatory model of experimental autoimmune encephalomyelitis (EAE), significant clinical and histopathological protective effects were evidenced, and these were related to an improved effector/regulatory cytokine balance in restimulated splenocytes. The essential role of curcumin-induced IL-10 for neuroprotection was confirmed by the complete abrogation of the clinical effects in IL-10-deficient mice. Finally, the translational therapeutic prospection of this strategy was evidenced by the neuroprotection observed in mice starting the treatment one week after disease triggering. Collectively, results demonstrate the power of a simple natural IL-10-inducing small molecule to tackle chronic inflammation, when its classical systemic and direct pharmacological view is shifted towards the targeting of regional immune cells, in order to rationally harness its immunopharmacological potential. This shift implies that many well-known IL-10-inducing small molecules could be easily reformulated and repurposed to develop safe, innovative, and accessible immune-based interventions for CID.
How to Synchronize Longitudinal Patient Data With the Underlying Disease Progression: A Pilot Study Using the Biomarker CRP for Timing COVID-19
Authors: Maibach Martina A. , Allam Ahmed , Hilty Matthias P. , Perez Gonzalez Nicolas A. , Buehler Philipp K. , Wendel Garcia Pedro D. , Brugger Silvio D. , Ganter Christoph C. ,   The CoViD-19 ICU-Research Group Zurich,   The RISC-19-ICU Investigators, Krauthammer Michael , Schuepbach Reto A. , Bartussek Jan
Published: July 2021

The continued digitalization of medicine has led to an increased availability of longitudinal patient data that allows the investigation of novel and known diseases in unprecedented detail. However, to accurately describe any underlying pathophysiology and allow inter-patient comparisons, individual patient trajectories have to be synchronized based on temporal markers. In this pilot study, we use longitudinal data from critically ill ICU COVID-19 patients to compare the commonly used alignment markers “onset of symptoms,” “hospital admission,” and “ICU admission” with a novel objective method based on the peak value of the inflammatory marker C-reactive protein (CRP). By applying our CRP-based method to align the progression of neutrophils and lymphocytes, we were able to define a pathophysiological window that improved mortality risk stratification in our COVID-19 patient cohort. Our data highlights that proper synchronization of longitudinal patient data is crucial for accurate interpatient comparisons and the definition of relevant subgroups. The use of objective temporal disease markers will facilitate both translational research efforts and multicenter trials.