bluuebunny/crossref_metadata_2025 huggingface dataset:
Documentation of Pharmaceutical Development. Part 1. Study of the Documentation of the Site
Authors: Smekhova I. E. , Shigarova L. V. , Meteleva V. D. , Flisyuk E. V.
Published: February 2021

Introduction. The trends in the modern pharmaceutical industry demonstrate the need for proper documentation of all stages of the life cycle of a medicinal product to ensure its quality and eliminate risks to patients. Therefore, documenting pharmaceutical development, as the first stage of this cycle, is relevant. It will allow regulating technological processes, product quality indicators, as well as ensuring the integrity of the data received. It is advisable to document the pharmaceutical development taking into account the requirements established at the enterprise where the industrial production of this drug is planned.

Aim. Development of documentation for pharmaceutical development within the pharmaceutical quality system of the enterprise. One of the objectives of the study was to analyze the document flow of the pharmaceutical quality system of the enterprise planning the production of suppositories for the proper development of documentation for the pharmaceutical development of a medicinal product in this dosage form.

Materials and methods. During the research the following methods were used: content analysis; system analysis; sociological (survey method); SWOT analysis technology; systems approach.

Results and discussion. In order to get acquainted with modern trends related to pharmaceutical development and pharmaceutical quality system, at the first stage, an analysis of the ICH (International Council on Harmonization) guidance document Quality Implementation Working Group on Q8, Q9 and Q10 Questions & Answers was carried out. An important preparatory stage for the optimal documentation of a pharmaceutical development was the study of the document flow of the pharmaceutical quality system of an enterprise planning to produce two-component suppositories. For this purpose, in the course of the research, a questionnaire was developed containing questions related to the functioning of the quality system, documentation processes and production. The analysis of the received responses made it possible to obtain information on documenting the pharmaceutical development of medicinal products, as well as to identify the main trends associated with this process within the pharmaceutical quality system.

Conclusion. The documentation of the pharmaceutical quality system of the enterprise planning the production of suppositories has been analyzed. The strengths and weaknesses of the documentation are identified, a number of proposals for improving the documentation system are highlighted. Based on the results obtained, proposals will be developed for the optimal documentation of pharmaceutical development and a set of documents for an enterprise planning to produce two-component suppositories will be formed.
Descrição epidemiológica da meningite no Nordeste brasileiro: casos notificados em 2019
Authors: Fontes Francisco Lucas de Lima , Silva Nayla Karine Barros da , Silva Hallyson Leno Lucas da , Marques Thalyta Maria Campelo , Barros Deylane de Melo , Oliveira Ilana Isla , Silva Denise Sabrina Nunes da , Silva Lucilene da Silva , Castro Mara Cléssia de Oliveira , Silva Camila Fernandes da , Silva Iraci de Sousa e , Lima Thiago Nascimento
Published: February 2021

Objetivo: Descrever os aspectos epidemiológicos da meningite no Nordeste brasileiro no ano de 2019. Métodos: Trata-se de um estudo descritivo, retrospectivo com abordagem quantitativa desenvolvido por meio do Sistema de Informação de Agravos de Notificação vinculado ao Departamento de Informática do Sistema Único de Saúde. As variáveis coletadas para a construção desta pesquisa foram: estado proveniente dos casos (com inclusão apenas dos casos da região Nordeste), sexo, raça/cor, faixa etária, etiologia dos casos confirmados e desfecho. Resultados: No ano investigado foram notificados 2.487 casos de meningite no Nordeste brasileiro. Dentre os estados da região, Pernambuco apresentou o maior número de casos, seguido do Ceará e da Bahia. Entre os indivíduos acometidos pela patologia evidenciou-se prevalência do sexo masculino em 60,64% dos casos. Houve maior manifestação em adultos jovens na faixa etária de 20 a 39 anos, o que representou 25,81% do total de casos. A alta foi vista em 76% dos desfechos. Observou-se a meningite viral como a forma mais prevalente, representando 38,16%. Conclusão: Os dados apresentados aqui possuem relevância. Mesmo com a existência de imunização oferecida gratuitamente pelo Sistema Único de Saúde para os agentes etiológicos mais comuns da meningite os casos na região Nordeste continuam a ocorrer. Torna-se relevante chamar a atenção também para a subnotificação, com existência de casos que não são reportados às autoridades competentes.
Enxerto de ramo mandibular para reconstrução maxilar em pacientes pós cirurgia ortognática: relato de caso
Authors: Costa Filho João Roberto Trindade , Rodrigues João Victor Soares , Siqueira Natália Barbosa de , Trindade Pedro Ivo Tavares , Monteiro Naara Gabriela , Pitol Leticia Palin , Silva Ana Cláudia Ervolino da , Hochuli-Vieira Eduardo , Vasconcelos Belmiro Cavalcanti do Egito
Published: February 2021

A ausência de tecido ósseo remanescente dos rebordos alveolares tem sido um grande problema para a reabilitação estética e funcional nos pacientes. O enxerto ósseo autógeno é o material ideal para reconstrução dos rebordos maxilares e mandibulares. A escolha da área doadora, seja intrabucal ou extrabucal, está relacionada com a quantidade e a qualidade do enxerto ósseo necessária para cada caso. O presente estudo tem por objetivo relatar um caso clínico de reconstrução maxilar em paciente após cirurgia ortognática utilizando-se como área doadora, a região de ramo mandibular, e revisar as principais características relacionadas à técnica cirúrgica.
Consulta de enfermagem sistematizada na atenção básica: relato de experiência
Authors: Azevedo Suely Lopes de , Parente Juliana da Silva , Souza Lorrany Viana de , Almeida Giulia Lemos de , Queiroz Priscila das Neves , Silva Jorge Luiz Lima da , Oliveira Aline Silva da Fonte Santa Rosa de
Published: February 2021

Objetivo: relatar as experiências da sistematização da assistência de enfermagem (SAE) junto aos usuários hipertensos, descrever as atividades assistenciais e educacionais durante a consulta de enfermagem. Método: trata-se de um relato de experiência sobre o uso da SAE na consulta de enfermagem realizada na unidade básica de saúde. Resultados: durante a consulta de enfermagem sistematizada foi estimulado o autocuidado, implementado escuta ativa e o plano assistencial e educacional como principais intervenções de enfermagem. Grande parte dos usuários passou a aderir ao tratamento medicamentoso e não medicamentoso, com significativa mudança de seu estilo de vida. Conclusão: A SAE apesar de ser o caminho para um atendimento organizado e de qualidade, é um grande desafio para a enfermagem. Esta oportunidade possibilitou a experiência da atuação do enfermeiro na atenção básica de saúde.
Resistance mechanisms of Cryptococcus spp. and plant compounds as tools to combat them
Authors: Silva Suéllen Pedrosa da , Costa Clarice Barbosa Lucena da , Silva José Dayvid Ferreira da , Alves Robson Raion de Vasconcelos , Silva Guilherme Antonio de Souza , Freitas Anderson Felipe Soares de , Guedes Caio César da Silva , Marinho Amanda de Oliveira , Paiva Patrícia Maria Guedes , Napoleão Thiago Henrique
Published: February 2021

Cryptococcus is a genus of dimorphic basidiomycete fungi found in the form of yeasts and filaments. Cryptococcosis has as main etiological agents the species Cryptococcus neoformans and Cryptococcus gattii. This disease is considered a public health problem and has becoming more alarming because of the limitations of antimicrobials available to its treatment, in addition to an increase in reports of fungal resistance. In this sense, the present review sought to survey information on the resistance mechanisms of Cryptococcus spp. against the main drugs used in cryptococcosis therapy as well as on the antimicrobial activities of plants against these fungi. Studies have reported that several mechanisms may be involved in fungal resistance to drugs including drug inactivation by enzymes, expression of efflux pumps and others drug transporters, as well as changes in the drug target and/or implementation of alternative metabolic pathways. As an alternative to conventional antimicrobials, substances and molecules extracted from plants have demonstrated potential for controlling these pathogens. These phytochemicals can trigger the inhibition and/or death of Cryptococcus through morphological changes on fungi cells, inhibition of ergosterol synthesis, cell leakage, capsular decrease, interference in cell division, reduction of activity of several enzymes such as laccase and urease, inhibition of biofilm formation, among others. In this sense, plants are an important source of bioactive compounds with antimicrobial activity that can be studied in the search for new drugs that are increasingly effective, specific and less toxic in the control of cryptococcosis.
in vitro and in silico studies of interaction of synthetic 2,6,9-trisubstituted purine kinase inhibitors BPA-302, BP-21 and BP-117 with liver drug-metabolizing cytochromes P450
Authors: Špičáková A , Kraus P , Gucký T , Kryštof V , Strnad M , Bazgier V , Otyepka M , Kubíčková V , Poruba M , Rácová Z , Zapletalová I , Anzenbacher P
Published: February 2021

An evaluation of possible interactions with enzymes of drug metabolism (cytochromes P450, CYP) is an important part of studies on safety and, in general, on the properties of any drug or biologically active compound. The article is focused on the preliminary metabolic study of selected 2,6,9-trisubstituted purine kinase inhibitors with significant anticancer activities which we have developed. The compounds BP-21 and BP-117 represent strong CDK inhibitors and the compound BPA-302 was developed as selective FLT3-ITD kinase inhibitor. Here, emphasis is placed on interactions of these compounds with the nine most important forms of CYP to evaluate the possibility of inhibition of these enzymes. The possibility of their inhibitory effect was studied in vitro on selected human liver microsomal CYP enzymes. The most affected enzyme was CYP2C19. Its activity dropped to 22 % of its original value by BPA 302, to 13 % by BP-21 and to 6 % by BP-117 at the highest concentration tested (250 µmol·l 1). The results suggest that the metabolism of concomitantly administered drugs should not be significantly affected at lower doses. Molecular docking of BPA-302 indicated that it can bind to active site of both CYP2C19 and CYP2D6 enzymes above the heme cofactor corroborating the experimental data.
Differential CircRNA Expression Signatures May Serve as Potential Novel Biomarkers in Prostate Cancer
Authors: Greene John , Baird Anne-Marie , Lim Marvin , Flynn Joshua , McNevin Ciara , Brady Lauren , Sheils Orla , Gray Steven G. , McDermott Raymond , Finn Stephen P.
Published: February 2021

Circular RNAs (circRNAs), a recently discovered non-coding RNA, have a number of functions including the regulation of miRNA expression. They have been detected in a number of malignancies including prostate cancer (PCa). The differential expression pattern of circRNAs associated with PCa and androgen receptor (AR) status was investigated in this study. circRNA profiling was performed using a high throughout microarray assay on a panel of prostate cell lines, which consisted of normal, benign, and malignant cells (n = 9). circRNAs were more commonly significantly up-regulated (p < 0.05) than downregulated in malignant cell lines (n = 3,409) vs. benign cell lines (n = 2,949). In a grouped analysis based on AR status, there were 2,127 down-regulated circRNAs in androgen independent cell lines compared to 2,236 in androgen dependent cell lines, thus identifying a potential circRNA signature reflective of androgen dependency. Through a bioinformatics approach, the parental genes associated with the top 10 differentially expressed circRNAs were identified such as hsa_circ_0064644, whose predicted parental gene target is RBMS3, and hsa_circ_0060539, whose predicted gene target is SDC4. Furthermore, we identified three circRNAs associated with the parental gene Caprin1 (hsa_circ_0021652, hsa_circ_0000288, and hsa_circ_0021647). Other studies have shown the importance of Caprin1 in PCa cell survival and drug resistance. Given the modified circRNA expression signatures identified here, these hypothesis generating results suggest that circRNAs may serve as potential putative diagnostic and predictive markers in PCa. However, further validation studies are required to assess the true potential of these markers in the clinical setting.
Leptin Receptor q223r Polymorphism Influences Clostridioides difficile Infection-Induced Neutrophil CXCR2 Expression in an Interleukin-1β Dependent Manner
Authors: Horrigan Olivia , Jose Shinsmon , Mukherjee Anindita , Sharma Divya , Huber Alexander , Madan Rajat
Published: February 2021

Neutrophils are key first-responders in the innate immune response to C. difficile infection (CDI) and play a central role in disease pathogenesis. Studies have clearly shown that tissue neutrophil numbers need to be tightly regulated for optimal CDI outcomes: while excessive colonic neutrophilia is associated with severe CDI, neutrophil depletion also results in worse outcomes. However, the biological mechanisms that control CDI-induced neutrophilia remain poorly defined. C-X-C chemokine receptor 2 (CXCR2) is a chemotactic receptor that is critical in neutrophil mobilization from bone marrow to blood and tissue sites. We have previously reported that a single nucleotide polymorphism (SNP) in leptin receptor (LEPR), present in up to 50% of people, influenced CDI-induced neutrophil CXCR2 expression and tissue neutrophilia. Homozygosity for mutant LEPR (i.e. RR genotype) was associated with higher CXCR2 expression and more tissue neutrophils. Here, we investigated the biological mechanisms that regulate neutrophil CXCR2 expression after CDI, and the influence of host genetics on this process. Our data reveal that: a) CXCR2 plays a key role in CDI-induced neutrophil extravasation from blood to colonic tissue; b) plasma from C. difficile-infected mice upregulated CXCR2 on bone marrow neutrophils; c) plasma from C. difficile-infected RR mice induced a higher magnitude of CXCR2 upregulation and had more IL-1β; and d) IL-1β neutralization reduced CXCR2 expression on bone marrow and blood neutrophils and their subsequent accrual to colonic tissue. In sum, our data indicate that IL-1β is a key molecular mediator that communicates between gastro-intestinal tract (i.e. site of CDI) and bone marrow (i.e. primary neutrophil reservoir) and regulates the intensity of CDI-induced tissue neutrophilia by modulating CXCR2 expression. Further, our studies highlight the importance of host genetics in affecting these innate immune responses and provide novel insights into the mechanisms by which a common SNP influences CDI-induced neutrophilia.
Exposure Elements in Disaster Databases and Availability for Local Scale Application: Case Study of Kuala Lumpur, Malaysia
Authors: Muhamad Nurfashareena , Arshad Siti Hasniza M. , Pereira Joy Jacqueline
Published: February 2021

Exposure elements in open-access disaster databases that are relevant to critical infrastructure and basic services in the Sendai Framework on Disaster Risk Reduction (SFDRR) were transformed into spatial data, to investigate the impact of flash flood hazards in Kuala Lumpur, Malaysia. In this era of big data and hyper-connectivity, the availability of open-access data on exposure elements across scales and systems is largely unknown. Information on exposure elements and hazard susceptibility provide important insights to enhance community resilience, to move away from merely managing disasters to managing the risk of disasters, in line with the SFDRR. The case study of Kuala Lumpur enabled an assessment of information availability in existing disaster databases and within the national system, to facilitate informed decision-making. Findings reveal that there are a total of 26 databases on the internet that provide information on disasters and related elements; of which 18 are global, three are regional and four provide information at the national scale. However, only ten databases are open access where the user is able to easily retrieve information while others provide a “view only option”. The coverage of exposure elements in disaster databases is very poor where only five databases carried such information; and it is not useful for local scale application. Thus, information was sought from multiple open data sources within the national system and transformed into spatial data, to develop an exposure element data inventory for the city. There are 509 exposure elements within Kuala Lumpur, covering 33 private and government hospitals and community clinics; 189 public and private schools and institutions higher education; 261 facilities that provide basic services; and 26 features that represent of social and economic aspects. The exposure elements, which is coherent with the SFDRR, benefits decision-making when overlain with existing flood hazard zones and susceptible areas. Moving forward, emerging hazards due to climate change will be evaluated to strengthen informed decision-making and build community resilience in the city. The empowerment of local level research has great potential to advance open sharing of information on disaster and climate risks in the region.
Do Molecular Profiles of Primary Versus Metastatic Radioiodine Refractory Differentiated Thyroid Cancer Differ?
Authors: Gomes-Lima Cristiane J. , Shobab Leila , Wu Di , Ylli Dorina , Bikas Athanasios , McCoy Matthew , Feldman Rebecca , Lee Wen , Rao Sarika N. , Jensen Kirk , Vasko Vasily , Castro Luiz Claudio , Jonklaas Jacqueline , Wartofsky Leonard , Burman Kenneth D.
Published: February 2021

Management of metastatic radioiodine refractory differentiated thyroid cancer (DTC) can be a therapeutic challenge. Generally, little is known about the paired molecular profile of the primary tumor and the metastases and whether they harbor the same genetic abnormalities. The present study compared the molecular profile of paired tumor specimens (primary tumor/metastatic sites) from patients with radioiodine refractory DTC in order to gain insight into a possible basis for resistance to radioiodine. Twelve patients with radioiodine refractory metastases were studied; median age at diagnosis of 61 years (range, 25–82). Nine patients had papillary TC (PTC), one had follicular TC (FTC), and two had Hürthle cell TC (HTC). Distant metastases were present in the lungs (n = 10), bones (n = 4), and liver (n = 1). The molecular profiling of paired tumors was performed with a panel of 592 genes for Next Generation Sequencing, RNA-sequencing, and immunohistochemistry. Digital microfluidic PCR was used to investigateTERTpromoter mutations. The genetic landscape of all paired sites comprisedBRAF,NRAS,HRAS,TP53,ATM,MUTYH,POLE, andNTRKgenes, includingBRAFandNTRKfusions.BRAFV600E was the most common point mutation in the paired specimens (5/12).TERTpromoter mutation C228T was detected in one case. PD-L1 expression at metastatic sites was highly positive (95%) for one patient with HTC. All specimens were stable for microsatellite instability testing, and the tumor mutation burden was low to intermediate. Therefore, the molecular profile of DTC primary and metastatic lesions can show heterogeneity, which may help explain some altered responses to therapeutic intervention.
Positive Effects of NPY1 Receptor Activation on Islet Structure Are Driven by Pancreatic Alpha- and Beta-Cell Transdifferentiation in Diabetic Mice
Authors: Lafferty Ryan A. , Tanday Neil , Moffett R. Charlotte , Reimann Frank , Gribble Fiona M. , Flatt Peter R. , Irwin Nigel
Published: February 2021

Enzymatically stable and specific neuropeptide Y1 receptor (NPYR1) agonists, such as sea lamprey PYY(1-36) (SL-PYY(1-36)), are believed to improve glucose regulation in diabetes by targeting pancreatic islets. In this study, streptozotocin (STZ) diabetic transgenic GluCreERT2;ROSA26-eYFP and Ins1Cre/+;Rosa26-eYFP mouse models have been used to study effects of sustained NPYR1 activation on islet cell composition and alpha- and beta-cell lineage transitioning. STZ induced a particularly severe form of diabetes in Ins1Cre/+;Rosa26-eYFP mice, but twice-daily administration (25 nmol/kg) of SL-PYY(1-36) for 11 days consistently improved metabolic status. Blood glucose was decreased (p < 0.05 - p < 0.001) and both fasted plasma and pancreatic insulin significantly increased by SL-PYY(1-36). In both GluCreERT2;ROSA26-eYFP and Ins1Cre/+; Rosa26-eYFP mice, STZ provoked characteristic losses (p < 0.05 - p < 0.001) of islet numbers, beta-cell and pancreatic islet areas together with increases in area and central islet location of alpha-cells. With exception of alpha-cell area, these morphological changes were fully, or partially, returned to non-diabetic control levels by SL-PYY(1-36). Interestingly, STZ apparently triggered decreased (p < 0.001) alpha- to beta-cell transition in GluCreERT2;ROSA26-eYFP mice, together with increased loss of beta-cell identity in Ins1Cre/+;Rosa26-eYFP mice, but both effects were significantly (p < 0.001) reversed by SL-PYY(1-36). SL-PYY(1-36) also apparently reduced (p < 0.05) beta- to alpha-cell conversion in Ins1Cre/+;Rosa26-eYFP mice and glucagon expressing alpha-cells in GluCreERT2;ROSA26-eYFP mice. These data indicate that islet benefits of prolonged NPY1R activation, and especially restoration of beta-cell mass, are observed irrespective of diabetes status, being linked to cell lineage alterations including transdifferentiation of alpha- to beta-cells.
Nerve Growth Factor: A Dual Activator of Noradrenergic and Cholinergic Systems of the Rat Ovary
Authors: Benitez Agustin , Riquelme Raul , del Campo Miguel , Araya Camila , Lara Hernan E.
Published: February 2021

The functioning of the ovary is influenced by the autonomic system (sympathetic and cholinergic intraovarian system) which contributes to the regulation of steroid secretion, follicular development, and ovulation. There is no information on the primary signal that activates both systems. The nerve growth factor (NGF) was the first neurotrophic factor found to regulate ovarian noradrenergic neurons and the cholinergic neurons in the central nervous system. The aim of this study was to determine whether NGF is one of the participating neurotrophic factors in the activation of the sympathetic and cholinergic system of the ovary in vivo and its role in follicular development during normal or pathological states. The administration of estradiol valerate (a polycystic ovary [PCO] phenotype model) increased norepinephrine (NE) (through an NGF-dependent mechanism) and acetylcholine (ACh) levels. Intraovarian exposure of rats for 28 days to NGF (by means of an osmotic minipump) increased the expression of tyrosine hydroxylase and acetylcholinesterase (AChE, the enzyme that degrades ACh) without affecting enzyme activity but reduced ovarian ACh levels. In vitro exposure of the ovary to NGF (100 ng/ml for 3 h) increased both choline acetyl transferase and vesicular ACh transporter expression in the ovary, with no effect in ACh level. In vivo NGF led to an anovulatory condition with the appearance of follicular cysts and decreased number of corpora lutea (corresponding to noradrenergic activation). To determine whether the predominance of a NE-induced polycystic condition after NGF is responsible for the PCO phenotype, rats were exposed to an intraovarian administration of carbachol (100 μM), a muscarinic cholinergic agonist not degraded by AChE. Decreased the number of follicular cysts and increased the number of corpora lutea, reinforcing that cholinergic activity of the ovary participates in controlling its functions. Although NGF increased the biosynthetic capacity for ACh, it was not available to act in the ovary. Hence, NGF also regulates the ovarian cholinergic system, implying that NGF is the main regulator of the dual autonomic control. These findings highlight the need for research in the treatment of PCO syndrome by modification of locally produced ACh as an in vivo regulator of follicular development.
Assessing Ecosystem Services in Mangroves: Insights from São Tomé Island (Central Africa)
Authors: Afonso Filipa , Félix Pedro M. , Chainho Paula , Heumüller Joshua A. , de Lima Ricardo F. , Ribeiro Filipe , Brito Ana C.
Published: February 2021

Mangroves are some of the most productive coastal systems on the planet and provide valuable ecosystem services (ES). They are especially important in threatened ecosystems and developing countries, where they are likely to have direct impacts on local communities. An approach based on ES allows assessing ecosystems across the domains of ecology, sociology and economy. This study focused on the evaluation of ES in mangroves and started by creating a comprehensive global list of mangrove ES based on the Millennium Ecosystem Assessment. These services were then quantified using the best available indicators for mangrove systems. The mangroves of Diogo Nunes, São João dos Angolares and Malanza, located in the São Tomé Island, were used to illustrate the challenges in applying ES indicators in this type of ecosystems. The obtained results confirmed that mangroves can provide important and diverse services. However, the high variability among mangrove systems affects their ability to deliver ES, requiring caution for the extrapolation across regions. This assessment emphasizes how the ES framework can be used as a tool to develop management plans that integrate conservation goals and human wellbeing.
Distribution of Japanese Eel Anguilla japonica Revealed by Environmental DNA
Authors: Kasai Akihide , Yamazaki Aya , Ahn Hyojin , Yamanaka Hiroki , Kameyama Satoshi , Masuda Reiji , Azuma Nobuyuki , Kimura Shingo , Karaki Tatsuro , Kurokawa Yuko , Yamashita Yoh
Published: February 2021

The abundance of Japanese eel Anguilla japonica has rapidly decreased in recent decades. Following a re-evaluation of the possibility of extinction, the Japanese Ministry of the Environment and the International Union for Conservation of Nature listed the Japanese eel as an endangered species in 2013 and 2014, respectively. However, their abundance and precise distribution have never been clarified owing to their nocturnality and difficulty in their capture. In this study, the distribution of Japanese eels was investigated by monitoring for environmental DNA (eDNA), a non-invasive and efficient detection method. A total of 365 water samples were collected from 265 rivers located throughout Japan. High concentrations of eDNA of Japanese eels were detected in rivers on the Pacific side, but were low in the Sea of Japan side. In particular, very little eDNA amplification was confirmed from Hokkaido and the north of the Sea of Japan. The eDNA distribution in Japanese rivers coincides with the transport of the larvae in the ocean, as estimated by numerical simulations. Generalized linear mixed models were developed to explain the distribution of eDNA concentrations. The total nitrogen concentration emerged as an important factor in the best model. These results indicate that the distribution of Japanese eel is mostly determined by the maritime larval transport, and their survival and growth depend on the abundance of food in the river. The findings of the present study are useful for the management of populations and in the conservation of Japanese eels.
Protein Phosphatase 2A Catalytic Subunit PP2A-1 Enhances Rice Resistance to Sheath Blight Disease
Authors: Lin Qiu Jun , Chu Jin , Kumar Vikranth , Yuan De Peng , Li Zhi Min , Mei Qiong , Xuan Yuan Hu
Published: February 2021

Rice (Oryza sativa) production is damaged to a great extent by sheath blight disease (ShB). However, the defense mechanism in rice against this disease is largely unknown. Previous transcriptome analysis identified a significantly induced eukaryotic protein phosphatase 2A catalytic subunit 1 (PP2A-1) after the inoculation of Rhizoctonia solani. Five genes encoding PP2A exist in rice genome, and these five genes are ubiquitously expressed in different tissues and stages. Inoculation of R. solani showed that the genome edited pp2a-1 mutants using the CRISPR/Cas9 were more susceptible to ShB than the wild-type control, but other PP2A gene mutants exhibited similar response to ShB compared to wild-type plants. In parallel, PP2A-1 expression level was higher in the activation tagging line, and PP2A-1 overexpression inhibited plant height and promoted the resistance to ShB. PP2A-1-GFP was localized in the cytoplasm and nucleus. In addition, R. solani-dependent induction kinetics of pathogen-related genes PBZ1 and PR1b was lower in pp2a-1 mutants but higher in PP2A-1 activation line compared to those in the wild-type. In conclusion, our analysis shows that PP2A-1 is a member of protein phosphatase, which regulates rice resistance to ShB. This result broadens the understanding of the defense mechanism against ShB and provides a potential target for rice breeding for disease resistance.
Bisulfite-Converted DNA Quantity Evaluation: A Multiplex Quantitative Real-Time PCR System for Evaluation of Bisulfite Conversion
Authors: Hong Sae Rom , Shin Kyoung-Jin
Published: February 2021

Bisulfite (BS) conversion, which includes a series of chemical reactions using bisulfite, is a prerequisite to most DNA methylation analysis methods, and thus is an essential step in the associated research process. Unfortunately, BS conversion leads to the degradation or loss of DNA, which can hinder further downstream analysis. In addition, it is well known that incomplete BS conversion is crucial, as it causes an exaggeration of the DNA methylation level, which can adversely affect the results. Therefore, there have been many attempts to measure three key features of BS conversion: BS conversion efficiency, recovery, and degradation level. In this study, a multiplex quantitative real-time PCR system named BisQuE was suggested to simultaneously analyze three important aspects of the conversion step. By adopting cytosine-free PCR primers for two differently sized multicopy regions, the short amplicon and long amplicon were obtained from both the genomic and BS-converted DNA, thus enabling the obtaining of reliable and sensitive results and the calculation of the degradation level of the conversion step. Also, probes for detecting converted/unconverted templates and C-T indicators for inducing the formula were included in this assay to quantify BS-converted DNA in order to compute the conversion efficiency and recovery. Six BS conversion kits (EZ DNA Methylation-Lightning Kit, Premium Bisulfite kit, MethylEdge® Bisulfite Conversion System, EpiJET Bisulfite Conversion Kit, EpiTect Fast DNA Bisulfite Kit, and NEBNext® Enzymatic Methyl-seq Conversion Module) were tested in 20 samples using 50 ng of genomic DNA as an input with the BisQuE. The conversion efficiency, degradation levels, as well as recovery rates of the kits were investigated. A total of 99.61–99.90% conversion efficiency was perceived for five of the kits, while the NEBNext kit showed about 94%. The lowest degradation level was shown by the NEBNext kit, whereas the other kits were quite similar. The recovery rates of the kits were found to be within the range of 18–50%. A Qubit assay was also used to compare the recovery rate of BisQuE.
Integrative Genomic and Transcriptomic Analyses of Tumor Suppressor Genes and Their Role on Tumor Microenvironment and Immunity in Lung Squamous Cell Carcinoma
Authors: Kim Ahreum , Lim Sun Min , Kim Joo-Hang , Seo Jeong-Sun
Published: February 2021

Non-small-cell lung cancers (NSCLCs) are largely classified into lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC), which have different therapeutic options according to its molecular profiles and immune checkpoint expression, especially PD-L1, which is a suppressive factor in the tumor microenvironment. The tumor microenvironment can be altered by the genomic mutations on specific innate immune genes as well as tumor suppressor genes, so it is essential to comprehend the association between tumor microenvironment and tumor suppressor genes to discover the promising immunotherapeutic strategy to overcome the resistance of immune check point blockade. In this study, we aimed to analyze how the somatic mutations in tumor suppressor genes affect the tumor immune microenvironment through a comprehensive analysis of mutational profiling on the representative tumor suppressor genes (TP53, CDKN2A, PTEN, RB1, BRCA1, BRCA2) and immune gene expression in The Cancer Genome Atlas (TCGA) 155 lung squamous cell carcinoma (LUSC) and 196 lung adenocarcinoma (LUAD) samples. Several microenvironmental factors, such as the infiltrating immune and stromal cells, were suppressed by the mutated tumor suppressor genes in LUSC, unlike in the LUAD samples. In particular, infiltrating immune cells such as macrophage, neutrophil, and dendritic cells were significantly reduced in tumors with mutated tumor suppressor genes’ group. In addition, the gene expressions for interleukin production and lymphocyte differentiation and PGC, C7, HGF, PLA2G2A, IL1RL1, CCR2, ALOX15B, CXCL11, FCN3 were significantly down-regulated, which were key immune genes for the cross-talk between LUSC microenvironment and tumor suppressors. Therefore, we generated evidence that TSG mutations in LUSC have an impact on tumor immune microenvironment, which suggests that TSG non-mutated patients will have the more inflamed tumors and are more likely to respond to immune checkpoint blockade therapy.
Tick-Tattoo: DNA Vaccination Against B. burgdorferi or Ixodes scapularis Tick Proteins
Authors: Klouwens Michelle J. , Trentelman Jos J. A. , Wagemakers Alex , Ersoz Jasmin I. , Bins Adriaan D. , Hovius Joppe W.
Published: February 2021

Introduction

Borrelia burgdorferi sensu lato (sl) is the causative agent of Lyme borreliosis. Currently there is no human vaccine against Lyme borreliosis, and most research focuses on recombinant protein vaccines. DNA tattoo vaccination with B. afzelii strain PKo OspC in mice has proven to be fully protective against B. afzelii syringe challenge and induces a favorable humoral immunity compared to recombinant protein vaccination. Alternatively, several recombinant protein vaccines based on tick proteins have shown promising effect in tick-bite infection models. In this study, we evaluated the efficacy of DNA vaccines against Borrelia OspC or tick antigens in a tick-bite infection model.

Method

We vaccinated C3H/HeN mice with OspC using a codon-optimized DNA vaccine or with recombinant protein. We challenged these mice with B. burgdorferi sensu stricto (ss)-infected Ixodes scapularis nymphs. Subsequently, we vaccinated C3H/HeN mice with DNA vaccines coding for tick proteins for which recombinant protein vaccines have previously resulted in interference with tick feeding and/or Borrelia transmission: Salp15, tHRF, TSLPI, and Tix-5. These mice were also challenged with B. burgdorferi ss infected Ixodes scapularis nymphs.

Results

DNA tattoo and recombinant OspC vaccination both induced total IgG responses. Borrelia cultures and DNA loads of skin and bladder remained negative in the mice vaccinated with OspC DNA vaccination, except for one culture. DNA vaccines against tick antigens Salp15 and Tix-5 induced IgG responses, while those against tHRF and TSLPI barely induced any IgG response. In addition, Borrelia cultures, and DNA loads from mice tattooed with DNA vaccines against tick proteins TSLPI, Salp15, tHRF, and Tix-5 were all positive.

Conclusion

A DNA tattoo vaccine against OspC induced high specific IgG titers and provided near total protection against B. burgdorferi ss infection by tick challenge. In contrast, DNA tattoo vaccines against tick proteins TSLPI, Salp15, tHRF, and Tix-5 induced low to moderate IgG titers and did not provide protection. Therefore, DNA tattoo vaccination does not seem a suitable vaccine strategy to identify, or screen for, tick antigens for anti-tick vaccines. However, DNA tattoo vaccination is a straightforward and effective vaccination platform to assess novel B. burgdorferi sl antigen candidates in a relevant tick challenge model.
Perspectives on Oncolytic Salmonella in Cancer Immunotherapy—A Promising Strategy
Authors: Wang Ding , Wei Xiaodong , Kalvakolanu Dhan V. , Guo Baofeng , Zhang Ling
Published: February 2021

Since the first reported spontaneous regression of tumors in patients withstreptococcusinfection, cancer biological therapy was born and it evolved into today’s immunotherapy over the last century. Although the original strategy was unable to impart maximal therapeutic benefit at the beginning, it laid the foundations for the development of immune checkpoint blockade and CAR-T which are currently used for cancer treatment in the clinics. However, clinical applications have shown that current cancer immunotherapy can cause a series of adverse reactions and are captious for patients with preexisting autoimmune disorders.Salmonellaewas first reported to exert antitumor effect in 1935. Until now, numerous studies have proved its potency as an antitumor agent in the near future. In this review, we summarize the currently available data on the antitumor effects ofSalmonella, and discussed a possibility of integratingSalmonellainto cancer immunotherapy to overcome current obstacles.
Preventing Atopic Diseases During Childhood – Early Exposure Matters
Authors: Pierau Mandy , Arra Aditya , Brunner-Weinzierl Monika C.
Published: February 2021

Atopic diseases in childhood are a major burden worldwide and there is still a lack of knowledge about treatable causes. In industrialized countries such as Germany, almost every second child is sensitized to at least one common allergen. Recent studies show that although the predisposition to allergies is inherited, the adaptive immune system of neonates and infants follows a developmental trajectory and whether an allergy actually occurs depends also on timing of allergen exposure including diet as well as environmental factors. New recommendations are far from being rigid of allergen avoidance; it is rather moving toward conditions that stand for more biodiversity. The observation that introduction of peanuts or eggs early in life significantly reduced the development of a later allergy will change our recommendations for the introduction of complementary foods. This is consistent with the hygiene hypothesis that early provocation shapes the developing immune system so that it reacts appropriately. Therefore, promoting the development of tolerance is at the heart of sensible allergy prevention - and this begins with the last trimester of pregnancy. In light of this concept, actual recommendations are discussed.
Parainfluenza Virus 5 Priming Followed by SIV/HIV Virus-Like-Particle Boosting Induces Potent and Durable Immune Responses in Nonhuman Primates
Authors: Xiao Peng , Dienger-Stambaugh Krista , Chen Xuemin , Wei Huiling , Phan Shannon , Beavis Ashley C. , Singh Karnail , Adhikary Nihar R. Deb , Tiwari Pooja , Villinger Francois , He Biao , Spearman Paul
Published: February 2021

The search for a preventive vaccine against HIV infection remains an ongoing challenge, indicating the need for novel approaches. Parainfluenza virus 5 (PIV5) is a paramyxovirus replicating in the upper airways that is not associated with any animal or human pathology. In animal models, PIV5-vectored vaccines have shown protection against influenza, RSV, and other human pathogens. Here, we generated PIV5 vaccines expressing HIV envelope (Env) and SIV Gag and administered them intranasally to macaques, followed by boosting with virus-like particles (VLPs) containing trimeric HIV Env. Moreover, we compared the immune responses generated by PIV5-SHIV prime/VLPs boost regimen in naïve vs a control group in which pre-existing immunity to the PIV5 vector was established. We demonstrate for the first time that intranasal administration of PIV5-based HIV vaccines is safe, well-tolerated and immunogenic, and that boosting with adjuvanted trimeric Env VLPs enhances humoral and cellular immune responses. The PIV5 prime/VLPs boost regimen induced robust and durable systemic and mucosal Env-specific antibody titers with functional activities including ADCC and neutralization. This regimen also induced highly polyfunctional antigen-specific T cell responses. Importantly, we show that diminished responses due to PIV5 pre-existing immunity can be overcome in part with VLP protein boosts. Overall, these results establish that PIV5-based HIV vaccine candidates are promising and warrant further investigation including moving on to primate challenge studies.
Apoptotic Donor Cells in Transplantation
Authors: Husain Irma , Luo Xunrong
Published: February 2021

Despite significant advances in prevention and treatment of transplant rejection with immunosuppressive medications, we continue to face challenges of long-term graft survival, detrimental medication side effects to both the recipient and transplanted organ together with risks for opportunistic infections. Transplantation tolerance has so far only been achieved through hematopoietic chimerism, which carries with it a serious and life-threatening risk of graft versus host disease, along with variability in persistence of chimerism and uncertainty of sustained tolerance. More recently, numerousin vitroandin vivostudies have explored the therapeutic potential of silent clearance of apoptotic cells which have been well known to aid in maintaining peripheral tolerance to self. Apoptotic cells from a donor not only have the ability of down regulating the immune response, but also are a way of providing donor antigens to recipient antigen-presenting-cells that can then promote donor-specific peripheral tolerance. Herein, we review both laboratory and clinical evidence that support the utility of apoptotic cell-based therapies in prevention and treatment of graftversushost disease and transplant rejection along with induction of donor-specific tolerance in solid organ transplantation. We have highlighted the potential limitations and challenges of this apoptotic donor cell-based therapy together with ongoing advancements and attempts made to overcome them.
The Factors Affecting Expansion of Reactive Tumor Infiltrating Lymphocytes (TIL) From Bladder Cancer and Potential Therapeutic Applications
Authors: Aydin Ahmet Murat , Bunch Brittany L. , Beatty Matthew , Hajiran Ali , Dhillon Jasreman , Sarnaik Amod A. , Pilon-Thomas Shari , Poch Michael A.
Published: February 2021

Tumor infiltrating lymphocytes (TIL) therapy was shown to provide durable objective response in patients with metastatic melanoma. As a fundamental first step to bring TIL therapy to clinical use, identification of patients whose tumors yield optimal numbers of reactive TIL is indispensable. We have previously shown that expansion of tumor reactive TIL from primary bladder tumors and lymph node metastases is feasible. Here, we performed TIL harvesting from additional surgical specimens (additional 31 primary tumors and 10 lymph nodes) to generate a heterogenous cohort of 53 patients with bladder cancer (BC) to evaluate the tumor characteristics that lead to tumor-reactive TIL expansion. Among a total of 53 patients, overall TIL growth from tumor samples were 37/53 (69.8%) and overall anti-tumor reactive TIL were 26/35 (74.3%). Mixed urothelial carcinoma is associated with higher anti-tumor reactivity of expanded TIL than pure urothelial carcinoma (89.5% vs. 56.3%, p=0.049). The anti-tumor reactivity of expanded TIL from primary tumors previously treated with BCG immunotherapy were lower (33.3% vs. 82.6%, p=0.027) although T-cell phenotype (CD3+, CD4+, CD8+, and CD56+) was similar regardless prior of BCG therapy. Addition of agonistic 4-1BB antibody in culture media with IL-2 improved the number of expanded TIL from primary tumors previously treated with BCG immunotherapy. There was no significant difference between basal and luminal subtype tumors in terms of viable and reactive TIL growth. Our study demonstrates that TIL expansion is feasible across all BC patients and BC subtypes, and we suggest that TIL therapy can be a reasonable treatment strategy for various manifestations of BC.
Superoxide Dismutase 3 Controls the Activation and Differentiation of CD4+T Cells
Authors: Agrahari Gaurav , Sah Shyam Kishor , Bang Chul Hwan , Kim Yeong Ho , Kim Tae-Yoon
Published: February 2021

Superoxide dismutase 3 (SOD3), a well-known antioxidant has been shown to possess immunomodulatory properties through inhibition of T cell differentiation. However, the underlying inhibitory mechanism of SOD3 on T cell differentiation is not well understood. In this study, we investigated the effect of SOD3 on anti-CD3/CD28- or phorbol myristate acetate (PMA) and ionomycin (ION)-mediated activation of mouse naive CD4+T cells. Our data showed that SOD3 suppressed the expression of activation-induced surface receptor proteins such as CD25, and CD69, and cytokines production. Similarly, SOD3 was found to reduce CD4+T cells proliferation and suppress the activation of downstream pathways such as ERK, p38, and NF-κB. Moreover, naïve CD4+T cells isolated from global SOD3 knock-out mice showed higher expression of CD25, CD69, and CD71, IL-2 production, proliferation, and downstream signals compared to wild-type CD4+T cells. Whereas, the use of DETCA, a known inhibitor of SOD3 activity, found to nullify the inhibitory effect of SOD3 on CD4+T cell activation of both SOD3 KO and wild-type mice. Furthermore, the expression of surface receptor proteins, IL-2 production, and downstream signals were also reduced in Th2 and Th17 differentiated cells upon SOD3 treatment. Overall, our data showed that SOD3 can attenuate CD4+T cell activation through modulation of the downstream signalings and restrict CD4+T cell differentiation. Therefore, SOD3 can be a promising therapeutic for T cell-mediated disorders.