PeerJ
type: channel
link: https://peerj.com/articles/
description: Articles published in PeerJ
publisher: PeerJ Inc.
publication name: PeerJ
issn: 2167-8359
items: Dissection of genetic architecture for tiller angle in rice (Oryza sativa. L) by multiple genome-wide association analyses
items: Genotypic variation in root architectural traits under contrasting phosphorus levels in Mediterranean and Indian origin lentil genotypes
items: A versatile isothermal amplification assay for the detection of leptospires from various sample types
items: Expression of IER3 in hepatocellular carcinoma: clinicopathology, prognosis, and potential regulatory pathways
items: First observations of ovary regeneration in an amphipod, Ampelisca eschrichtii Krøyer, 1842
items: Evaluation of the diagnostic and prognostic values of serum HSP90α in sepsis patients: a retrospective study
items: Effect of magnification factor by Galilean loupes on working posture of dental students in simulated clinical procedures: associations between direct and observational measurements
items: Functional analysis of ARF1 from Cymbidium goeringii in IAA response during leaf development
items: Effect of precipitation change on the photosynthetic performance of Phragmites australis under elevated temperature conditions
items: Feature screening for survival trait with application to TCGA high-dimensional genomic data

Dissection of genetic architecture for tiller angle in rice (Oryza sativa. L) by multiple genome-wide association analyses
type: item
type: AcademicArticle
description: Background The rice plant architecture is determined by spatially and temporally domesticated tiller angle. The deeper insight into the genetic mechanism for rice plant architecture will allow more efficient light capture by increasing the planting density, reproducibility, and the ability to survive in a stressful environment. Methods In this study, a natural population of 795 genotypes further divided into japonica and indica subpopulations, was evaluated for tiller angle. A significant variation with a wide range was observed. Genome-wide association analysis was performed by the general linear model (GLM), and compressed mix linear model (cMLM) for three populations to disclose the genomic associations. The population principal components and kinship matrix in 1,000 permutations were used to remove the false positives. The candidate genes were evaluated for their functional annotations and specific molecular pathways. The sequencing-based haplotype analysis was further performed to reveal the functional variation among candidate genomic regions. Results As a result, 37 significant QTLs with 93 annotated loci were identified. Among the loci, a known tiller angle controlling locus TAC1 was also identified. The introduction of the sequence pooling technique was observed fruitful to screen the 12 significant QTLs with 22 annotated loci. For ten of these loci, the functional variations were identified by haplotype analysis. These results were not only providing a better understanding of the genetic bases of rice plant architecture but also provide significant information for future breeding programs.
format: application/pdf
format: text/html
creator: Muhammad Abdul Rehman Rashid
creator: Rana Muhammad Atif
creator: Yan Zhao
creator: Farrukh Azeem
creator: Hafiz Ghulam Muhu-Din Ahmed
creator: Yinghua Pan
creator: Danting Li
creator: Yong Zhao
creator: Zhanying Zhang
creator: Hongliang Zhang
creator: Jinjie Li
creator: Zichao Li
subject: Agricultural Science
subject: Genomics
subject: Plant Science
description: doi:10.7717/peerj.12674
publisher: PeerJ Inc.
rights: © 2022 Rashid et al.
abstract: Background The rice plant architecture is determined by spatially and temporally domesticated tiller angle. The deeper insight into the genetic mechanism for rice plant architecture will allow more efficient light capture by increasing the planting density, reproducibility, and the ability to survive in a stressful environment. Methods In this study, a natural population of 795 genotypes further divided into japonica and indica subpopulations, was evaluated for tiller angle. A significant variation with a wide range was observed. Genome-wide association analysis was performed by the general linear model (GLM), and compressed mix linear model (cMLM) for three populations to disclose the genomic associations. The population principal components and kinship matrix in 1,000 permutations were used to remove the false positives. The candidate genes were evaluated for their functional annotations and specific molecular pathways. The sequencing-based haplotype analysis was further performed to reveal the functional variation among candidate genomic regions. Results As a result, 37 significant QTLs with 93 annotated loci were identified. Among the loci, a known tiller angle controlling locus TAC1 was also identified. The introduction of the sequence pooling technique was observed fruitful to screen the 12 significant QTLs with 22 annotated loci. For ten of these loci, the functional variations were identified by haplotype analysis. These results were not only providing a better understanding of the genetic bases of rice plant architecture but also provide significant information for future breeding programs.
bibliographic citation: Rashid et al. (2022) Dissection of genetic architecture for tiller angle in rice (Oryza sativa. L) by multiple genome-wide association analyses. PeerJ 10:e12674 https://doi.org/10.7717/peerj.12674
date submitted: 2021-08-12
date accepted: 2021-12-02
issued: 2022-03-10
is part of: PeerJ
license: https://creativecommons.org/licenses/by/4.0/
author list: Agent
  family name: Rashid
  given name: Muhammad Abdul Rehman
author list: Agent
  family name: Atif
  given name: Rana Muhammad
author list: Agent
  family name: Zhao
  given name: Yan
author list: Agent
  family name: Azeem
  given name: Farrukh
author list: Agent
  family name: Ahmed
  given name: Hafiz Ghulam Muhu-Din
author list: Agent
  family name: Pan
  given name: Yinghua
author list: Agent
  family name: Li
  given name: Danting
author list: Agent
  family name: Zhao
  given name: Yong
author list: Agent
  family name: Zhang
  given name: Zhanying
author list: Agent
  family name: Zhang
  given name: Hongliang
author list: Agent
  family name: Li
  given name: Jinjie
author list: Agent
  family name: Li
  given name: Zichao
doi: 10.7717/peerj.12674
volume: 10
issn: 2167-8359
publication name: PeerJ
publication date: 2022-03-10
copyright: © 2022 Rashid et al.
attribution name: Rashid et al.

Genotypic variation in root architectural traits under contrasting phosphorus levels in Mediterranean and Indian origin lentil genotypes
type: item
type: AcademicArticle
description: The development of phosphorus-efficient crop cultivars boosts productivity while lowering eutrophication in the environment. It is feasible to improve the efficiency of phosphorus (P) absorption in lentils by enhancing phosphorus absorption through root architectural traits. The root architectural traits of 110 diverse lentil genotypes of Indian and Mediterranean origin were assessed, and the relationships between traits were investigated. In a hydroponics experiment, the lentil lines were examined at the seedling stage under two conditions: adequate P supply and deficient P supply. The Pearson correlation coefficients between root architectural traits and genetic diversity among lentil lines were assessed. To estimate variance components, a model (fixed factor) was used. In this experiment, both phosphorus (P) and genotype were fixed variables. Our lentil lines showed significant genetic variability and considerable genetic diversity for all traits under both treatments. The TRL (total root length) and PRL (primary root length) showed strong positive associations with all other characteristics excluding root average diameter (RAD) in both P treatments. In both P treatments, the RAD revealed a negative significant association with Total Root Tips (TRT), as well as total root volume (TRV) and total root forks (TRF) in the deficit conditions of P. Total root volume (TRV), total surface area (TSA), and total root tips had higher coefficient variance values. The first two principal components represented 67.88% and 66.19% of the overall variance in the adequate and deficit P treatments respectively. The Shannon-Weaver diversity index (H′) revealed that RAD, PRL, and TSA had more variability than TRT and TRF under both treatments. According to the Comprehensive Phosphorus Efficiency Measure (CPEM), the best five highly efficient genotypes are PLL 18-09, PLS 18-01, PLL 18-25, PLS 18-23, and PLL 18-07, while IG112131, P560206, IG334, L11-231, and PLS18-67 are highly ineffi…
format: application/pdf
format: text/html
creator: Muraleedhar Aski
creator: Reena Mehra
creator: Gyan Prakash Mishra
creator: Dharmendra Singh
creator: Prachi Yadav
creator: Neha Rai
creator: Venkata Ravi Prakash Reddy
creator: Arun Kumar MB
creator: Renu Pandey
creator: Madan Pal Singh
creator: Gayacharan
creator: Ruchi Bansal
creator: Kuldeep Tripathi
creator: Sripada M. Udupa
creator: Shiv Kumar
creator: Ashutosh Sarker
creator: Harsh Kumar Dikshit
subject: Agricultural Science
subject: Plant Science
subject: Natural Resource Management
description: doi:10.7717/peerj.12766
publisher: PeerJ Inc.
rights: © 2022 Aski et al.
abstract: The development of phosphorus-efficient crop cultivars boosts productivity while lowering eutrophication in the environment. It is feasible to improve the efficiency of phosphorus (P) absorption in lentils by enhancing phosphorus absorption through root architectural traits. The root architectural traits of 110 diverse lentil genotypes of Indian and Mediterranean origin were assessed, and the relationships between traits were investigated. In a hydroponics experiment, the lentil lines were examined at the seedling stage under two conditions: adequate P supply and deficient P supply. The Pearson correlation coefficients between root architectural traits and genetic diversity among lentil lines were assessed. To estimate variance components, a model (fixed factor) was used. In this experiment, both phosphorus (P) and genotype were fixed variables. Our lentil lines showed significant genetic variability and considerable genetic diversity for all traits under both treatments. The TRL (total root length) and PRL (primary root length) showed strong positive associations with all other characteristics excluding root average diameter (RAD) in both P treatments. In both P treatments, the RAD revealed a negative significant association with Total Root Tips (TRT), as well as total root volume (TRV) and total root forks (TRF) in the deficit conditions of P. Total root volume (TRV), total surface area (TSA), and total root tips had higher coefficient variance values. The first two principal components represented 67.88% and 66.19% of the overall variance in the adequate and deficit P treatments respectively. The Shannon-Weaver diversity index (H′) revealed that RAD, PRL, and TSA had more variability than TRT and TRF under both treatments. According to the Comprehensive Phosphorus Efficiency Measure (CPEM), the best five highly efficient genotypes are PLL 18-09, PLS 18-01, PLL 18-25, PLS 18-23, and PLL 18-07, while IG112131, P560206, IG334, L11-231, and PLS18-67 are highly ineffi…
bibliographic citation: Aski et al. (2022) Genotypic variation in root architectural traits under contrasting phosphorus levels in Mediterranean and Indian origin lentil genotypes. PeerJ 10:e12766 https://doi.org/10.7717/peerj.12766
date submitted: 2021-07-07
date accepted: 2021-12-17
issued: 2022-03-10
is part of: PeerJ
license: https://creativecommons.org/licenses/by/4.0/
author list: Agent
  family name: Aski
  given name: Muraleedhar
author list: Agent
  family name: Mehra
  given name: Reena
author list: Agent
  family name: Mishra
  given name: Gyan Prakash
author list: Agent
  family name: Singh
  given name: Dharmendra
author list: Agent
  family name: Yadav
  given name: Prachi
author list: Agent
  family name: Rai
  given name: Neha
author list: Agent
  family name: Reddy
  given name: Venkata Ravi Prakash
author list: Agent
  family name: MB
  given name: Arun Kumar
author list: Agent
  family name: Pandey
  given name: Renu
author list: Agent
  family name: Singh
  given name: Madan Pal
author list: Agent
  family name: Gayacharan
author list: Agent
  family name: Bansal
  given name: Ruchi
author list: Agent
  family name: Tripathi
  given name: Kuldeep
author list: Agent
  family name: Udupa
  given name: Sripada M.
author list: Agent
  family name: Kumar
  given name: Shiv
author list: Agent
  family name: Sarker
  given name: Ashutosh
author list: Agent
  family name: Dikshit
  given name: Harsh Kumar
doi: 10.7717/peerj.12766
volume: 10
issn: 2167-8359
publication name: PeerJ
publication date: 2022-03-10
copyright: © 2022 Aski et al.
attribution name: Aski et al.

A versatile isothermal amplification assay for the detection of leptospires from various sample types
type: item
type: AcademicArticle
description: Background Leptospirosis is a zoonotic disease caused by bacteria of the genus Leptospira that affects both humans and animals worldwide. Early detection of the pathogen in humans is crucial for early intervention and control of the progression of the disease to a severe state. It is also vitally important to be able to detect the presence of the pathogen in carrier animals to control the spread of the disease from the environment. Here we developed a simple and rapid loop-mediated isothermal amplification (LAMP) assay targeting the leptospiral secY gene. Results Several reaction conditions of the LAMP reaction were optimized to ensure efficient amplification of the target DNA. The sensitivity of the developed LAMP assay obtained using a pure Leptospira culture was 2 × 104 copies of genomic DNA per reaction (equivalent to 0.1 ng) for a 40-minute reaction time. No cross-reactions were observed in the LAMP reaction against a series of non-leptospiral bacteria, indicating a specific reaction. The applicability of the LAMP assay was demonstrated on human blood and urine specimens collected from suspected leptospirosis patients and rat kidney specimens collected from suspected leptospirosis outbreak areas and high-risk areas. The developed LAMP assay demonstrated a higher detection rate for leptospiral DNA compared with the polymerase chain reaction (PCR) assay, possibly due to the presence of inhibitory substances, especially in rat kidney specimens, to which the PCR method is more susceptible. The present findings also highlight the importance of urine sample collection from patients for routine monitoring of the disease. Conclusions In short, the developed LAMP assay can serve as a feasible alternative tool for the diagnosis of leptospirosis and be used for epidemiological and environmental surveillance of the disease, considering its robustness, rapidity, sensitivity, and specificity, as demonstrated in this study.
format: application/pdf
format: text/html
creator: Shuhaidah Othman
creator: Pui-Yuei Lee
creator: Jia-Yong Lam
creator: Noraini Philip
creator: Nurul Natasya Azhari
creator: Norliza Bahtiar Affendy
creator: Siti Norbaya Masri
creator: Vasantha Kumari Neela
creator: Farah Shafawati Mohd-Taib
creator: Hui-Yee Chee
subject: Microbiology
subject: Molecular Biology
description: doi:10.7717/peerj.12850
publisher: PeerJ Inc.
rights: © 2022 Othman et al.
abstract: Background Leptospirosis is a zoonotic disease caused by bacteria of the genus Leptospira that affects both humans and animals worldwide. Early detection of the pathogen in humans is crucial for early intervention and control of the progression of the disease to a severe state. It is also vitally important to be able to detect the presence of the pathogen in carrier animals to control the spread of the disease from the environment. Here we developed a simple and rapid loop-mediated isothermal amplification (LAMP) assay targeting the leptospiral secY gene. Results Several reaction conditions of the LAMP reaction were optimized to ensure efficient amplification of the target DNA. The sensitivity of the developed LAMP assay obtained using a pure Leptospira culture was 2 × 104 copies of genomic DNA per reaction (equivalent to 0.1 ng) for a 40-minute reaction time. No cross-reactions were observed in the LAMP reaction against a series of non-leptospiral bacteria, indicating a specific reaction. The applicability of the LAMP assay was demonstrated on human blood and urine specimens collected from suspected leptospirosis patients and rat kidney specimens collected from suspected leptospirosis outbreak areas and high-risk areas. The developed LAMP assay demonstrated a higher detection rate for leptospiral DNA compared with the polymerase chain reaction (PCR) assay, possibly due to the presence of inhibitory substances, especially in rat kidney specimens, to which the PCR method is more susceptible. The present findings also highlight the importance of urine sample collection from patients for routine monitoring of the disease. Conclusions In short, the developed LAMP assay can serve as a feasible alternative tool for the diagnosis of leptospirosis and be used for epidemiological and environmental surveillance of the disease, considering its robustness, rapidity, sensitivity, and specificity, as demonstrated in this study.
bibliographic citation: Othman et al. (2022) A versatile isothermal amplification assay for the detection of leptospires from various sample types. PeerJ 10:e12850 https://doi.org/10.7717/peerj.12850
date submitted: 2021-10-05
date accepted: 2022-01-07
issued: 2022-03-10
is part of: PeerJ
license: https://creativecommons.org/licenses/by/4.0/
author list: Agent
  family name: Othman
  given name: Shuhaidah
author list: Agent
  family name: Lee
  given name: Pui-Yuei
author list: Agent
  family name: Lam
  given name: Jia-Yong
author list: Agent
  family name: Philip
  given name: Noraini
author list: Agent
  family name: Azhari
  given name: Nurul Natasya
author list: Agent
  family name: Affendy
  given name: Norliza Bahtiar
author list: Agent
  family name: Masri
  given name: Siti Norbaya
author list: Agent
  family name: Neela
  given name: Vasantha Kumari
author list: Agent
  family name: Mohd-Taib
  given name: Farah Shafawati
author list: Agent
  family name: Chee
  given name: Hui-Yee
doi: 10.7717/peerj.12850
volume: 10
issn: 2167-8359
publication name: PeerJ
publication date: 2022-03-10
copyright: ©2022 Othman et al.
attribution name: Othman et al.

Expression of IER3 in hepatocellular carcinoma: clinicopathology, prognosis, and potential regulatory pathways
type: item
type: AcademicArticle
description: Background Immediate early response 3 (IER3) is correlated to the prognosis of several cancers, but the precise mechanisms underlying the regulation by IER3 of the occurrence and development of hepatocellular carcinoma (HCC) remain unknown. Methods The expression level of IER3 was examined by using in-house immunohistochemistry (IHC), public gene chip, and public RNA-sequencing (RNA-seq). The standardized mean difference (SMD) was calculated to compare the expression levels of IER3 between HCC patients and controls. The summary receiver operating characteristics (sROC) was plotted to comprehensively understand the discriminatory capability of IER3 between HCC and non-HCC group. The Kaplan–Meier curves and the combined hazard ratios (HRs) were used to determine the prognostic value of IER3 in HCC. Moreover, differentially expressed genes (DEGs) and co-expression genes (CEGs) were used to explored the molecular mechanisms of IER3 underlying HCC. hTFtarget was used to predict the transcription factors (TFs) of IER3. The binding site of TFs and the IER3 promoter region was forecasted using the JASPAR website. The relevant ChIP-seq data were used to determine whether TF peaks were present in the IER3 transcription initiation. Results A significantly increased expression of IER3 protein was found in HCC tissue relative to non-HCC tissue as detected by IHC (p < 0.001). Compared to 1,263 cases of non-HCC tissues, IER3 in 1483 cases of HCC tissues was upregulated (SMD = 0.42, 95% confidence interval [CI] [0.09–0.76]). The sROC showed that IER3 had a certain ability at differentiating HCC tissues (area under the curve (AUC) = 0.65, 95% CI [0.61–0.69]). Comprehensive analysis of the effect of IER3 on the prognosis of patients with HCC demonstrated that higher IER3 expression was associated with poor prognosis in HCC (HRs = 1.30, 95% CI [1.03–1.64]). Pathway enrichment analysis revealed that IER3-related genes were mostly enriched in the PI3K-Akt signaling pathway, cancer-relat…
format: application/pdf
format: text/html
creator: Fei-Yan He
creator: Gang Chen
creator: Rong-quan He
creator: Zhi-Guang Huang
creator: Jian-Di Li
creator: Wei-Zi Wu
creator: Ji-Tian Chen
creator: Yu-Lu Tang
creator: Dong-Ming Li
creator: Shang-Ling Pan
creator: Zhen-Bo Feng
creator: Yi-wu Dang
subject: Bioinformatics
subject: Molecular Biology
subject: Gastroenterology and Hepatology
subject: Oncology
description: doi:10.7717/peerj.12944
publisher: PeerJ Inc.
rights: © 2022 He et al.
abstract: Background Immediate early response 3 (IER3) is correlated to the prognosis of several cancers, but the precise mechanisms underlying the regulation by IER3 of the occurrence and development of hepatocellular carcinoma (HCC) remain unknown. Methods The expression level of IER3 was examined by using in-house immunohistochemistry (IHC), public gene chip, and public RNA-sequencing (RNA-seq). The standardized mean difference (SMD) was calculated to compare the expression levels of IER3 between HCC patients and controls. The summary receiver operating characteristics (sROC) was plotted to comprehensively understand the discriminatory capability of IER3 between HCC and non-HCC group. The Kaplan–Meier curves and the combined hazard ratios (HRs) were used to determine the prognostic value of IER3 in HCC. Moreover, differentially expressed genes (DEGs) and co-expression genes (CEGs) were used to explored the molecular mechanisms of IER3 underlying HCC. hTFtarget was used to predict the transcription factors (TFs) of IER3. The binding site of TFs and the IER3 promoter region was forecasted using the JASPAR website. The relevant ChIP-seq data were used to determine whether TF peaks were present in the IER3 transcription initiation. Results A significantly increased expression of IER3 protein was found in HCC tissue relative to non-HCC tissue as detected by IHC (p < 0.001). Compared to 1,263 cases of non-HCC tissues, IER3 in 1483 cases of HCC tissues was upregulated (SMD = 0.42, 95% confidence interval [CI] [0.09–0.76]). The sROC showed that IER3 had a certain ability at differentiating HCC tissues (area under the curve (AUC) = 0.65, 95% CI [0.61–0.69]). Comprehensive analysis of the effect of IER3 on the prognosis of patients with HCC demonstrated that higher IER3 expression was associated with poor prognosis in HCC (HRs = 1.30, 95% CI [1.03–1.64]). Pathway enrichment analysis revealed that IER3-related genes were mostly enriched in the PI3K-Akt signaling pathway, cancer-relat…
bibliographic citation: He et al. (2022) Expression of IER3 in hepatocellular carcinoma: clinicopathology, prognosis, and potential regulatory pathways. PeerJ 10:e12944 https://doi.org/10.7717/peerj.12944
date submitted: 2021-08-27
date accepted: 2022-01-24
issued: 2022-03-10
is part of: PeerJ
license: https://creativecommons.org/licenses/by/4.0/
author list: Agent
  family name: He
  given name: Fei-Yan
author list: Agent
  family name: Chen
  given name: Gang
author list: Agent
  family name: He
  given name: Rong-quan
author list: Agent
  family name: Huang
  given name: Zhi-Guang
author list: Agent
  family name: Li
  given name: Jian-Di
author list: Agent
  family name: Wu
  given name: Wei-Zi
author list: Agent
  family name: Chen
  given name: Ji-Tian
author list: Agent
  family name: Tang
  given name: Yu-Lu
author list: Agent
  family name: Li
  given name: Dong-Ming
author list: Agent
  family name: Pan
  given name: Shang-Ling
author list: Agent
  family name: Feng
  given name: Zhen-Bo
author list: Agent
  family name: Dang
  given name: Yi-wu
doi: 10.7717/peerj.12944
volume: 10
issn: 2167-8359
publication name: PeerJ
publication date: 2022-03-10
copyright: ©2022 He et al.
attribution name: He et al.

First observations of ovary regeneration in an amphipod, Ampelisca eschrichtii Krøyer, 1842
type: item
type: AcademicArticle
description: Background Females of the gammaridean amphipod Ampelisca eschrichtii with signs of regenerating, previously atrophied ovaries were recovered from the northeastern shelf of Sakhalin Island (Okhotsk Sea, Russia). Ovarian regeneration was previously unknown for any amphipod species. A. eschrichtii have a predominantly 2-year life cycle (from embryo to adult death) and reproduce once between late winter or early spring at the age of 2 years. Occasionally, females survive to a third year. An adaptive value of extended survival among these females is likely to require that they are also reproductive. Methods Histological sections from a second-year female with ovarian atrophy, a female with normal ovaries, a third-year female with ovarian regeneration, as well as testes of an immature and a sexually mature male were compared to determine the sources of cells of the germinal and somatic lines necessary for ovarian regeneration. Results Ovarian regeneration in the third-year female began with the formation of a new germinal zone from germ cells preserved in the atrophied ovaries and eosinophilic cells of the previously starving second-year female. Eosinophilic cells form the mesodermal component of the germinal zone. A mass of these cells appeared in the second-year female that had atrophied ovaries and in large numbers on the intestine wall of the third-year female with regenerating ovaries. These eosinophilic cells appear to migrate into the regenerating ovaries. Conclusions All germ cells of the second-year female are not lost during ovarian atrophy and can be involved in subsequent ovarian regeneration. Eosinophilic cells involved in ovarian regeneration are of mesodermal origin. The eosinophilic cell morphologies are similar to those of quiescence cells (cells in a reversible state that do not divide but retain the ability to re-enter cell division and participate in regeneration). These histological data thus indicate that eosinophilic and germ cells of third-year fem…
format: application/pdf
format: text/html
creator: Valentina B. Durkina
creator: John W. Chapman
creator: Natalia L. Demchenko
subject: Conservation Biology
subject: Developmental Biology
subject: Ecology
subject: Marine Biology
subject: Histology
description: doi:10.7717/peerj.12950
publisher: PeerJ Inc.
rights: © 2022 Durkina et al.
abstract: Background Females of the gammaridean amphipod Ampelisca eschrichtii with signs of regenerating, previously atrophied ovaries were recovered from the northeastern shelf of Sakhalin Island (Okhotsk Sea, Russia). Ovarian regeneration was previously unknown for any amphipod species. A. eschrichtii have a predominantly 2-year life cycle (from embryo to adult death) and reproduce once between late winter or early spring at the age of 2 years. Occasionally, females survive to a third year. An adaptive value of extended survival among these females is likely to require that they are also reproductive. Methods Histological sections from a second-year female with ovarian atrophy, a female with normal ovaries, a third-year female with ovarian regeneration, as well as testes of an immature and a sexually mature male were compared to determine the sources of cells of the germinal and somatic lines necessary for ovarian regeneration. Results Ovarian regeneration in the third-year female began with the formation of a new germinal zone from germ cells preserved in the atrophied ovaries and eosinophilic cells of the previously starving second-year female. Eosinophilic cells form the mesodermal component of the germinal zone. A mass of these cells appeared in the second-year female that had atrophied ovaries and in large numbers on the intestine wall of the third-year female with regenerating ovaries. These eosinophilic cells appear to migrate into the regenerating ovaries. Conclusions All germ cells of the second-year female are not lost during ovarian atrophy and can be involved in subsequent ovarian regeneration. Eosinophilic cells involved in ovarian regeneration are of mesodermal origin. The eosinophilic cell morphologies are similar to those of quiescence cells (cells in a reversible state that do not divide but retain the ability to re-enter cell division and participate in regeneration). These histological data thus indicate that eosinophilic and germ cells of third-year fem…
bibliographic citation: Durkina et al. (2022) First observations of ovary regeneration in an amphipod, Ampelisca eschrichtii Krøyer, 1842. PeerJ 10:e12950 https://doi.org/10.7717/peerj.12950
date submitted: 2021-08-27
date accepted: 2022-01-26
issued: 2022-03-10
is part of: PeerJ
license: https://creativecommons.org/licenses/by/4.0/
author list: Agent
  family name: Durkina
  given name: Valentina B.
author list: Agent
  family name: Chapman
  given name: John W.
author list: Agent
  family name: Demchenko
  given name: Natalia L.
doi: 10.7717/peerj.12950
volume: 10
issn: 2167-8359
publication name: PeerJ
publication date: 2022-03-10
copyright: © 2022 Durkina et al.
attribution name: Durkina et al.

Evaluation of the diagnostic and prognostic values of serum HSP90α in sepsis patients: a retrospective study
type: item
type: AcademicArticle
description: Background Sepsis is a serious syndrome that is caused by immune responses dysfunction and leads to high mortality. The abilities of heat shock protein 90α (HSP90α) in assessing the diagnosis and prognosis in patients with sepsis remain ill-defined to date. We conducted a study to reveal the possible clinical applications of HSP90α as biomarker for the diagnosis and prognosis in patients with sepsis. Methods In total, 150 patients of sepsis, 110 patients without sepsis admitted to ICU and 110 healthy subjects were involved in this study. The serum HSP90α contents, sequential organ failure assessment (SOFA) scores, procalcitonin (PCT), and short-term survival status of the participants were measured and compared. Logistic and linear regression models adjusting for potential confounders were used to examine the association of HSP90α with sepsis survival. Moreover, serum IL-1β, IL-18, MIP-3α, and ENA-78 were also determined. Finally, Spearman correlation analysis was employed to reveal a possible mechanism that HSP90α contributed to the short-term deaths. Results Serum HSP90α levels in sepsis patients were higher than those in ICU controls and healthy controls (P < 0.001), and even increased in patients who died within 28 days (P < 0.001). Logistic and linear regression models identified HSP90α was an independent risk factors for sepsis mortality. Receiver operating characteristic (ROC) analysis displayed that HSP90α had a considerable predictive performance for sepsis outcome, with an area under curve (AUC) value up to 0.79. Survival analysis demonstrated that the mortality of sepsis individuals at 28 days was positively associated with HSP90α levels, especially the levels of HSP90α were greater than 120 ng/mL (P < 0.001). Moreover, among sepsis patients, those who died had notably elevated cytokines, IL-1β, IL-18, and chemokines, MIP-3α, ENA-78, relative to survivors. Further correlation analysis demonstrated that there was a nominally positive correlation between HS…
format: application/pdf
format: text/html
creator: Fuxing Li
creator: Yulin Zhang
creator: Bocheng Yu
creator: Zihua Zhang
creator: Yujuan Fan
creator: Li Wang
creator: Mingjing Cheng
creator: Ping Yan
creator: Weidong Zhao
subject: Emergency and Critical Care
subject: Hematology
subject: Infectious Diseases
subject: Translational Medicine
description: doi:10.7717/peerj.12997
publisher: PeerJ Inc.
rights: © 2022 Li et al.
abstract: Background Sepsis is a serious syndrome that is caused by immune responses dysfunction and leads to high mortality. The abilities of heat shock protein 90α (HSP90α) in assessing the diagnosis and prognosis in patients with sepsis remain ill-defined to date. We conducted a study to reveal the possible clinical applications of HSP90α as biomarker for the diagnosis and prognosis in patients with sepsis. Methods In total, 150 patients of sepsis, 110 patients without sepsis admitted to ICU and 110 healthy subjects were involved in this study. The serum HSP90α contents, sequential organ failure assessment (SOFA) scores, procalcitonin (PCT), and short-term survival status of the participants were measured and compared. Logistic and linear regression models adjusting for potential confounders were used to examine the association of HSP90α with sepsis survival. Moreover, serum IL-1β, IL-18, MIP-3α, and ENA-78 were also determined. Finally, Spearman correlation analysis was employed to reveal a possible mechanism that HSP90α contributed to the short-term deaths. Results Serum HSP90α levels in sepsis patients were higher than those in ICU controls and healthy controls (P < 0.001), and even increased in patients who died within 28 days (P < 0.001). Logistic and linear regression models identified HSP90α was an independent risk factors for sepsis mortality. Receiver operating characteristic (ROC) analysis displayed that HSP90α had a considerable predictive performance for sepsis outcome, with an area under curve (AUC) value up to 0.79. Survival analysis demonstrated that the mortality of sepsis individuals at 28 days was positively associated with HSP90α levels, especially the levels of HSP90α were greater than 120 ng/mL (P < 0.001). Moreover, among sepsis patients, those who died had notably elevated cytokines, IL-1β, IL-18, and chemokines, MIP-3α, ENA-78, relative to survivors. Further correlation analysis demonstrated that there was a nominally positive correlation between HS…
bibliographic citation: Li et al. (2022) Evaluation of the diagnostic and prognostic values of serum HSP90α in sepsis patients: a retrospective study. PeerJ 10:e12997 https://doi.org/10.7717/peerj.12997
date submitted: 2021-10-26
date accepted: 2022-02-02
issued: 2022-03-10
is part of: PeerJ
license: https://creativecommons.org/licenses/by/4.0/
author list: Agent
  family name: Li
  given name: Fuxing
author list: Agent
  family name: Zhang
  given name: Yulin
author list: Agent
  family name: Yu
  given name: Bocheng
author list: Agent
  family name: Zhang
  given name: Zihua
author list: Agent
  family name: Fan
  given name: Yujuan
author list: Agent
  family name: Wang
  given name: Li
author list: Agent
  family name: Cheng
  given name: Mingjing
author list: Agent
  family name: Yan
  given name: Ping
author list: Agent
  family name: Zhao
  given name: Weidong
doi: 10.7717/peerj.12997
volume: 10
issn: 2167-8359
publication name: PeerJ
publication date: 2022-03-10
copyright: © 2022 Li et al.
attribution name: Li et al.

Effect of magnification factor by Galilean loupes on working posture of dental students in simulated clinical procedures: associations between direct and observational measurements
type: item
type: AcademicArticle
description: Objectives To determine the effect of different levels of Galilean loupe magnification on working posture as measured by compliance with ergonomic posture positions, angular deviation from the neutral position of the neck, and muscle activation in the neck and upper back region during simulated clinical conditions. Methods An experimental laboratory study was performed in which the dependent variables were compliance with ergonomic posture requirements while performing simulated restorative procedures in Restorative Dentistry, angular deviation from the neutral position of the neck, and muscle activation in the neck and upper back. The independent variable was the level of Galilean loupe magnification, which was tested at four levels (naked eye, 2.5× magnification, 3.0× magnification, and 3.5× magnification). The cavity preparations and Class I composite resin restorations were performed on artificial first molars on a mannequin in a dental chair. The Compliance Assessment of Dental Ergonomic Posture Requirements (CADEP) was used for the postural analysis; as was an analysis of the angular deviation from the neutral position of the neck and surface electromyography. Working posture was recorded on video over the course of the procedure. Participants were filmed from three different angles. The Compliance Assessment of Dental Ergonomic Posture Requirements (CADEP) assessed compliance with ergonomic posture requirements. A locally produced posture assessment software analyzed angular deviation. Surface electromyography bilaterally assessed activation of the sternocleidomastoid, descending trapezius and ascending trapezius muscles. Two-factor analysis of variance (ANOVA) and either Tukey’s post-hoc test or the Games-Howell post-hoc test were performed (α = 0.05). Results During the cavity preparations and restorations, the use of Galilean loupes at all magnifications positively influenced working posture as measured by participants’ compliance with ergonomic posture po…
format: application/pdf
format: text/html
creator: Júlia Margato Pazos
creator: Simone Cecilio Hallak Regalo
creator: Paulo de Vasconcelos
creator: Juliana Alvares Duarte Bonini Campos
creator: Patricia Petromilli Nordi Sasso Garcia
subject: Dentistry
subject: Science and Medical Education
description: doi:10.7717/peerj.13021
publisher: PeerJ Inc.
rights: © 2022 Pazos et al.
abstract: Objectives To determine the effect of different levels of Galilean loupe magnification on working posture as measured by compliance with ergonomic posture positions, angular deviation from the neutral position of the neck, and muscle activation in the neck and upper back region during simulated clinical conditions. Methods An experimental laboratory study was performed in which the dependent variables were compliance with ergonomic posture requirements while performing simulated restorative procedures in Restorative Dentistry, angular deviation from the neutral position of the neck, and muscle activation in the neck and upper back. The independent variable was the level of Galilean loupe magnification, which was tested at four levels (naked eye, 2.5× magnification, 3.0× magnification, and 3.5× magnification). The cavity preparations and Class I composite resin restorations were performed on artificial first molars on a mannequin in a dental chair. The Compliance Assessment of Dental Ergonomic Posture Requirements (CADEP) was used for the postural analysis; as was an analysis of the angular deviation from the neutral position of the neck and surface electromyography. Working posture was recorded on video over the course of the procedure. Participants were filmed from three different angles. The Compliance Assessment of Dental Ergonomic Posture Requirements (CADEP) assessed compliance with ergonomic posture requirements. A locally produced posture assessment software analyzed angular deviation. Surface electromyography bilaterally assessed activation of the sternocleidomastoid, descending trapezius and ascending trapezius muscles. Two-factor analysis of variance (ANOVA) and either Tukey’s post-hoc test or the Games-Howell post-hoc test were performed (α = 0.05). Results During the cavity preparations and restorations, the use of Galilean loupes at all magnifications positively influenced working posture as measured by participants’ compliance with ergonomic posture po…
bibliographic citation: Pazos et al. (2022) Effect of magnification factor by Galilean loupes on working posture of dental students in simulated clinical procedures: associations between direct and observational measurements. PeerJ 10:e13021 https://doi.org/10.7717/peerj.13021
date submitted: 2021-08-02
date accepted: 2022-02-07
issued: 2022-03-10
is part of: PeerJ
license: https://creativecommons.org/licenses/by/4.0/
author list: Agent
  family name: Pazos
  given name: Júlia Margato
author list: Agent
  family name: Regalo
  given name: Simone Cecilio Hallak
author list: Agent
  family name: de Vasconcelos
  given name: Paulo
author list: Agent
  family name: Campos
  given name: Juliana Alvares Duarte Bonini
author list: Agent
  family name: Garcia
  given name: Patricia Petromilli Nordi Sasso
doi: 10.7717/peerj.13021
volume: 10
issn: 2167-8359
publication name: PeerJ
publication date: 2022-03-10
copyright: © 2022 Pazos et al.
attribution name: Pazos et al.

Functional analysis of ARF1 from Cymbidium goeringii in IAA response during leaf development
type: item
type: AcademicArticle
description: Background Cymbidium is an economically important genus of flowering orchids cultivated in China because of showing graceful leaf shapes and elegant flower coloration. However, the deterioration of the ecological environment and the difficulty of conservation management have become increasing challenges for maintaining its germplasm resources. ARFs are critical transcription factors in the auxin signaling pathway and have been found to play pivotal roles in leaf growth and development in previous studies. However, their functions and mechanisms in Cymbidium goeringii remain to be clarified. Methods The sequence of the CgARF1 gene was analyzed by bioinformatics. The expression of this gene in different tissues and under IAA treatment was detected by quantitative real-time PCR analysis. The CgARF1 gene was overexpressed in wild-type Arabidopsis and Nicotiana benthamiana via the Agrobacterium infection method. Acetone-ethanol solvent extraction was applied for the determination of chlorophyll contents, and the contents of endogenous hormones were determined using the enzyme-linked immunosorbent assay technique. Results CgARF1 cloned from C. goeringii ‘Songmei’ was 2,049 bp in length and encoded 682 amino acids containing three typical domains: a B3 DNA binding domain, an Aux_resp domain and an AUX/IXX family domain. The expression pattern of CgARF1 in different tissues of C. goeringii showed that its expression was highest in the leaves and changed greatly under IAA treatment. Subcellular localization studies showed that the protein was mainly localized in the cell nucleus. CgARF1-overexpressing lines exhibited leaf senescence and a decreased chlorophyll content. Under IAA treatment, CgARF1 regulates the rooting length, rooting number and rooting rate from detached leaves. The levels of endogenous hormones in transgenic leaves were also significantly changed. Conclusion These results indicated that CgARF1 overexpression is responsive to IAA treatment during leaf develo…
format: application/pdf
format: text/html
creator: Zihan Xu
creator: Fangle Li
creator: Meng Li
creator: Yuanhao He
creator: Yue Chen
creator: Fengrong Hu
subject: Developmental Biology
subject: Genetics
subject: Molecular Biology
subject: Plant Science
description: doi:10.7717/peerj.13077
publisher: PeerJ Inc.
rights: © 2022 Xu et al.
abstract: Background Cymbidium is an economically important genus of flowering orchids cultivated in China because of showing graceful leaf shapes and elegant flower coloration. However, the deterioration of the ecological environment and the difficulty of conservation management have become increasing challenges for maintaining its germplasm resources. ARFs are critical transcription factors in the auxin signaling pathway and have been found to play pivotal roles in leaf growth and development in previous studies. However, their functions and mechanisms in Cymbidium goeringii remain to be clarified. Methods The sequence of the CgARF1 gene was analyzed by bioinformatics. The expression of this gene in different tissues and under IAA treatment was detected by quantitative real-time PCR analysis. The CgARF1 gene was overexpressed in wild-type Arabidopsis and Nicotiana benthamiana via the Agrobacterium infection method. Acetone-ethanol solvent extraction was applied for the determination of chlorophyll contents, and the contents of endogenous hormones were determined using the enzyme-linked immunosorbent assay technique. Results CgARF1 cloned from C. goeringii ‘Songmei’ was 2,049 bp in length and encoded 682 amino acids containing three typical domains: a B3 DNA binding domain, an Aux_resp domain and an AUX/IXX family domain. The expression pattern of CgARF1 in different tissues of C. goeringii showed that its expression was highest in the leaves and changed greatly under IAA treatment. Subcellular localization studies showed that the protein was mainly localized in the cell nucleus. CgARF1-overexpressing lines exhibited leaf senescence and a decreased chlorophyll content. Under IAA treatment, CgARF1 regulates the rooting length, rooting number and rooting rate from detached leaves. The levels of endogenous hormones in transgenic leaves were also significantly changed. Conclusion These results indicated that CgARF1 overexpression is responsive to IAA treatment during leaf develo…
bibliographic citation: Xu et al. (2022) Functional analysis of ARF1 from Cymbidium goeringii in IAA response during leaf development. PeerJ 10:e13077 https://doi.org/10.7717/peerj.13077
date submitted: 2021-11-18
date accepted: 2022-02-16
issued: 2022-03-10
is part of: PeerJ
license: https://creativecommons.org/licenses/by/4.0/
author list: Agent
  family name: Xu
  given name: Zihan
author list: Agent
  family name: Li
  given name: Fangle
author list: Agent
  family name: Li
  given name: Meng
author list: Agent
  family name: He
  given name: Yuanhao
author list: Agent
  family name: Chen
  given name: Yue
author list: Agent
  family name: Hu
  given name: Fengrong
doi: 10.7717/peerj.13077
volume: 10
issn: 2167-8359
publication name: PeerJ
publication date: 2022-03-10
copyright: © 2022 Xu et al.
attribution name: Xu et al.

Effect of precipitation change on the photosynthetic performance of Phragmites australis under elevated temperature conditions
type: item
type: AcademicArticle
description: Background As a fundamental metabolism, leaf photosynthesis not only provides necessary energy for plant survival and growth but also plays an important role in global carbon fixation. However, photosynthesis is highly susceptible to environmental stresses and can be significantly influenced by future climate change. Methods In this study, we examined the photosynthetic responses of Phragmites australis (P. australis) to three precipitation treatments (control, decreased 30%, and increased 30%) under two thermal regimes (ambient temperature and +4 °C) in environment-controlled chambers. Results Our results showed that the net CO2 assimilation rate (Pn), maximal rate of Rubisco (Vcmax), maximal rate of ribulose-bisphosphate (RuBP) regeneration (Jmax) and chlorophyll (Chl) content were enhanced under increased precipitation condition, but were declined drastically under the condition of water deficit. The increased precipitation had no significant effect on malondialdehyde (MDA) content (p > 0.05), but water deficit drastically enhanced the MDA content by 10.1%. Meanwhile, a high temperature inhibited the positive effects of increased precipitation, aggravated the adverse effects of drought. The combination of high temperature and water deficit had more detrimental effect on P. australis than a single factor. Moreover, non-stomatal limitation caused by precipitation change played a major role in determining carbon assimilation rate. Under ambient temperature, Chl content had close relationship with Pn (R2 = 0.86, p < 0.01). Under high temperature, Pn was ralated to MDA content (R2 = 0.81, p < 0.01). High temperature disrupted the balance between Vcmax and Jmax (the ratio of Jmax to Vcmax decreased from 1.88 to 1.12) which resulted in a negative effect on the photosynthesis of P. australis. Furthermore, by the analysis of Chl fluorescence, we found that the xanthophyll cycle-mediated thermal dissipation played a major role in PSII photoprotection, resulting in no signi…
format: application/pdf
format: text/html
creator: Linhong Teng
creator: Hanyu Liu
creator: Xiaonan Chu
creator: Xiliang Song
creator: Lianhui Shi
subject: Agricultural Science
subject: Ecology
subject: Plant Science
description: doi:10.7717/peerj.13087
publisher: PeerJ Inc.
rights: © 2022 Teng et al.
abstract: Background As a fundamental metabolism, leaf photosynthesis not only provides necessary energy for plant survival and growth but also plays an important role in global carbon fixation. However, photosynthesis is highly susceptible to environmental stresses and can be significantly influenced by future climate change. Methods In this study, we examined the photosynthetic responses of Phragmites australis (P. australis) to three precipitation treatments (control, decreased 30%, and increased 30%) under two thermal regimes (ambient temperature and +4 °C) in environment-controlled chambers. Results Our results showed that the net CO2 assimilation rate (Pn), maximal rate of Rubisco (Vcmax), maximal rate of ribulose-bisphosphate (RuBP) regeneration (Jmax) and chlorophyll (Chl) content were enhanced under increased precipitation condition, but were declined drastically under the condition of water deficit. The increased precipitation had no significant effect on malondialdehyde (MDA) content (p > 0.05), but water deficit drastically enhanced the MDA content by 10.1%. Meanwhile, a high temperature inhibited the positive effects of increased precipitation, aggravated the adverse effects of drought. The combination of high temperature and water deficit had more detrimental effect on P. australis than a single factor. Moreover, non-stomatal limitation caused by precipitation change played a major role in determining carbon assimilation rate. Under ambient temperature, Chl content had close relationship with Pn (R2 = 0.86, p < 0.01). Under high temperature, Pn was ralated to MDA content (R2 = 0.81, p < 0.01). High temperature disrupted the balance between Vcmax and Jmax (the ratio of Jmax to Vcmax decreased from 1.88 to 1.12) which resulted in a negative effect on the photosynthesis of P. australis. Furthermore, by the analysis of Chl fluorescence, we found that the xanthophyll cycle-mediated thermal dissipation played a major role in PSII photoprotection, resulting in no signi…
bibliographic citation: Teng et al. (2022) Effect of precipitation change on the photosynthetic performance of Phragmites australis under elevated temperature conditions. PeerJ 10:e13087 https://doi.org/10.7717/peerj.13087
date submitted: 2021-10-26
date accepted: 2022-02-17
issued: 2022-03-10
is part of: PeerJ
license: https://creativecommons.org/licenses/by/4.0/
author list: Agent
  family name: Teng
  given name: Linhong
author list: Agent
  family name: Liu
  given name: Hanyu
author list: Agent
  family name: Chu
  given name: Xiaonan
author list: Agent
  family name: Song
  given name: Xiliang
author list: Agent
  family name: Shi
  given name: Lianhui
doi: 10.7717/peerj.13087
volume: 10
issn: 2167-8359
publication name: PeerJ
publication date: 2022-03-10
copyright: © 2022 Teng et al.
attribution name: Teng et al.

Feature screening for survival trait with application to TCGA high-dimensional genomic data
type: item
type: AcademicArticle
description: Background In high-dimensional survival genomic data, identifying cancer-related genes is a challenging and important subject in the field of bioinformatics. In recent years, many feature screening approaches for survival outcomes with high-dimensional survival genomic data have been developed; however, few studies have systematically compared these methods. The primary purpose of this article is to conduct a series of simulation studies for systematic comparison; the second purpose of this article is to use these feature screening methods to further establish a more accurate prediction model for patient survival based on the survival genomic datasets of The Cancer Genome Atlas (TCGA). Results Simulation studies prove that network-adjusted feature screening measurement performs well and outperforms existing popular univariate independent feature screening methods. In the application of real data, we show that the proposed network-adjusted feature screening approach leads to more accurate survival prediction than alternative methods that do not account for gene-gene dependency information. We also use TCGA clinical survival genetic data to identify biomarkers associated with clinical survival outcomes in patients with various cancers including esophageal, pancreatic, head and neck squamous cell, lung, and breast invasive carcinomas. Conclusions These applications reveal advantages of the new proposed network-adjusted feature selection method over alternative methods that do not consider gene-gene dependency information. We also identify cancer-related genes that are almost detected in the literature. As a result, the network-based screening method is reliable and credible.
format: application/pdf
format: text/html
creator: Jie-Huei Wang
creator: Cai-Rong Li
creator: Po-Lin Hou
subject: Bioinformatics
subject: Statistics
description: doi:10.7717/peerj.13098
publisher: PeerJ Inc.
rights: © 2022 Wang et al.
abstract: Background In high-dimensional survival genomic data, identifying cancer-related genes is a challenging and important subject in the field of bioinformatics. In recent years, many feature screening approaches for survival outcomes with high-dimensional survival genomic data have been developed; however, few studies have systematically compared these methods. The primary purpose of this article is to conduct a series of simulation studies for systematic comparison; the second purpose of this article is to use these feature screening methods to further establish a more accurate prediction model for patient survival based on the survival genomic datasets of The Cancer Genome Atlas (TCGA). Results Simulation studies prove that network-adjusted feature screening measurement performs well and outperforms existing popular univariate independent feature screening methods. In the application of real data, we show that the proposed network-adjusted feature screening approach leads to more accurate survival prediction than alternative methods that do not account for gene-gene dependency information. We also use TCGA clinical survival genetic data to identify biomarkers associated with clinical survival outcomes in patients with various cancers including esophageal, pancreatic, head and neck squamous cell, lung, and breast invasive carcinomas. Conclusions These applications reveal advantages of the new proposed network-adjusted feature selection method over alternative methods that do not consider gene-gene dependency information. We also identify cancer-related genes that are almost detected in the literature. As a result, the network-based screening method is reliable and credible.
bibliographic citation: Wang et al. (2022) Feature screening for survival trait with application to TCGA high-dimensional genomic data. PeerJ 10:e13098 https://doi.org/10.7717/peerj.13098
date submitted: 2021-09-29
date accepted: 2022-02-21
issued: 2022-03-10
is part of: PeerJ
license: https://creativecommons.org/licenses/by/4.0/
author list: Agent
  family name: Wang
  given name: Jie-Huei
author list: Agent
  family name: Li
  given name: Cai-Rong
author list: Agent
  family name: Hou
  given name: Po-Lin
doi: 10.7717/peerj.13098
volume: 10
issn: 2167-8359
publication name: PeerJ
publication date: 2022-03-10
copyright: © 2022 Wang et al.
attribution name: Wang et al.

PeerJ
type: Periodical
uri: PeerJ
issn: 2167-8359