bluuebunny/crossref_metadata_2025 huggingface dataset:
Radiation-induced abscopal effect and its enhancement by programmed cell death 1 blockade in the hepatocellular carcinoma: A murine model study
Authors: Yoo Gyu Sang , Ahn Won-Gyun , Kim Shin-Yeong , Kang Wonseok , Choi Changhoon , Park Hee Chul
Published: December 2020

Background/Aims: The abscopal effect, a rare phenomenon induced by radiation, can be reinforced by immunotherapy. Although radiation therapy and immunotherapy are increasingly being utilized for the treatment of hepatocellular carcinoma (HCC), whether immunotherapy could boost the abscopal effect remains unclear. In this study, we aimed to elucidate the immunological mechanisms underlying the abscopal effect induced by the combination of irradiation and immunotherapy in a murine HCC model.Methods: A syngeneic HCC mouse model was established by transplanting murine Hepa 1–6 HCC cells into both hind legs of immunocompetent C57BL/6 mice. The tumors on the right hind legs were irradiated, and abscopal effects were observed in the non-irradiated tumors on the left hind leg with or without the coadministration of anti-programmed cell death 1 (PD-1) antibodies. Flow cytometric analyses were performed to analyze the distributions of immune cells infiltrating both irradiated and non-irradiated tumors and the tumor-draining lymph nodes (TDLNs).Results: Administration of 16 Gy in two fractions more effectively inhibited the growth of both irradiated and nonirradiated tumors with higher tumor infiltration of cytotoxic T cells than 8 Gy did in a single fraction. The higher dose also increased activated dendritic cells in TDLNs, which had higher expression of the programmed cell death ligand 1. Coadministration of anti-PD-1 antibodies significantly enhanced the abscopal effect and increased infiltration of activated cytotoxic T cells in both irradiated and non-irradiated tumors.Conclusions: Our findings show that adding anti-PD-1 therapy to radiation enhanced the abscopal effect in a syngeneic murine model of HCC.
“Living like Queens”
Authors: Sergidou Katerina
Published: December 2020

This article focuses on the feminist mobilization that has characterized Cádiz Carnival since 2011, leading to the elimination of the Ninfas y Diosas (Nymphs and Goddesses) custom, a variant of the Reina de las Fiestas (Queen of Traditional Fiestas) ceremony introduced under Francisco Franco’s dictatorship (1939–75). By calling into question the representation of women in Carnival celebrations, female festive organizations have challenged the old, male-dominated festival traditions and transformed Cádiz Carnival. Their activism has carried over into everyday life, as female Carnival groups have created their own community and translated the artistic manifestations of their desire for equality into public policy. Using oral testimonies and archival material gathered during ethnographic fieldwork in the city, I trace the history of the reina and ninfas customs and analyze a variety of material related to their birth, evolution, and recent discontinuation. The ultimate purpose of this article is to map the tensions embedded in both the festival and contemporary Spanish society and to show how the Carnival stage can become a space where embodied feminist counter-hegemony is performed, thus contributing to the slow democratization of Spanish society.
Carnival against the Capital of Capital
Authors: Hammond John L.
Published: December 2020

The medieval carnival, according to Russian literary scholar Mikhail Bakhtin, was a public festivity of excess in which people were free to violate social norms and subvert prevailing authority. Recent analysts have applied Bakhtin’s concept of carnival to contemporary political protests that incorporate a playful, culture-defying element. But the term has been used in multiple and contradictory ways. For Bakhtin, carnival is an expressive pattern pervasive in a culture and has no instrumental purpose (what I call “communal carnival”), while carnivalesque protest consists of specific practices with an explicit political agenda (“intentional carnival”). The Occupy Wall Street movement can be analyzed as both communal and intentional carnival. Protest movements use humor to subvert received doctrines; humorous performances are addressed to participants, the public, and repressive forces. Some critics regard carnivalesque performances as frivolous and demeaning of serious political causes. I conclude by discussing the effect of carnival on the Occupy movement.
Å utvikle yrkeskompetanse gjennom arbeidslivspraksis i helsesekretærutdanningen
Authors: Johnsen Kjersti Lisbeth , Hiim Hilde
Published: December 2020

This article presents findings from a three-year research and development project where a high school and a hospital collaborated on the education of medical secretaries. The aim was to strengthen the medical secretary students’ development of vocational competence through close  collaboration between the school and the hospital on practice periods for the students. The results are based  on qualitative data from observations of the students during practice, observations of students when experiences from practice were presented at school, group interviews with students, teachers and instructors, and analyses of students’ reflection logs. The findings show that cooperation between the parties to define key functions and tasks in the work of medical secretaries, and to structure practice periods around these tasks, contributed to the students getting a great learning benefit from practice. The students expressed that they understood what the vocation was about, began to identify with it and felt the need to learn theory. However, they wanted better coherence between learning in practice and at school. The results suggest that mandatory practice periods and common structure for learning content in practice and at school should be main principles in the new curriculum for education of medical secretaries from autumn 2020.
Nanoparticles for Stem Cell Therapy Bioengineering in Glioma
Authors: Ruiz-Garcia Henry , Alvarado-Estrada Keila , Krishnan Sunil , Quinones-Hinojosa Alfredo , Trifiletti Daniel M.
Published: December 2020

Gliomas are a dismal disease associated with poor survival and high morbidity. Current standard treatments have reached a therapeutic plateau even after combining maximal safe resection, radiation, and chemotherapy. In this setting, stem cells (SCs) have risen as a promising therapeutic armamentarium, given their intrinsic tumor homing as well as their natural or bioengineered antitumor properties. The interplay between stem cells and other therapeutic approaches such as nanoparticles holds the potential to synergize the advantages from the combined therapeutic strategies. Nanoparticles represent a broad spectrum of synthetic and natural biomaterials that have been proven effective in expanding diagnostic and therapeutic efforts, either used alone or in combination with immune, genetic, or cellular therapies. Stem cells have been bioengineered using these biomaterials to enhance their natural properties as well as to act as their vehicle when anticancer nanoparticles need to be delivered into the tumor microenvironment in a very precise manner. Here, we describe the recent developments of this new paradigm in the treatment of malignant gliomas.
Tumor Targeted Multifunctional Magnetic Nanobubbles for MR/US Dual Imaging and Focused Ultrasound Triggered Drug Delivery
Authors: Jin Zhen , Chang Jinlong , Dou Peipei , Jin Shang , Jiao Min , Tang Heyun , Jiang Wenshuai , Ren Wu , Zheng Shaohui
Published: December 2020

The development of multifunctional nanoplatforms that are safe and have multiple therapeutic functions integrated with dual- or multi-imaging modality is one of the most urgent medical requirements for active cancer therapy. In our study, we prepared multifunctional magnetic nanobubbles (MF-MNBs) by co-encapsulating superparamagnetic iron oxide nanoparticles (SPIONs) and doxorubicin into polylactideco–glycolide–polyethylene glycol–folate (PLGA-PEG-FA) polymer-based nanobubbles for tumor-targeted ultrasound (US)/magnetic resonance (MR) imaging and focused ultrasound (FUS)-triggered drug delivery. Hydrophobic SPIONs were successfully embedded into MF-MNBs by a typical double emulsion process. The MF-MNBs were highly dispersed with well-defined spherical morphology and an average diameter of 208.4 ± 12.58 nm. The potential of MF-MNB as a dual-modal contrast agent for US and MR imaging was investigated via in vitro study, and the MF-MNB exhibits promising US/MR contrast ability. Moreover, tumor targeting ability was further enhanced by folate conjugation and assessed through in vitro cell test. Furthermore, FUS, as a non-invasive and remote-control technique, was adopted to trigger the release of doxorubicin from MF-MNB and generate the sonoporation effect to enhance drug release and cellular uptake of MF-MNBs. The 4T1 cell viability was significantly decreased by FA ligand-receptor-mediated targeting and FUS sonication. In addition, the developed MF-MNB also exhibits enhanced accumulation in tumor site by FA ligand-receptor-mediated tumor targeting, in which the accumulation of MF-MNB was further enhanced by FUS sonication. Hence, we believe that the MF-MNB could be a promising drug nanocarrier for US/MR-guided anticancer drug delivery to improve cancer treatment efficacy.
A Systematic Review of the Associations Between Inverse Dynamics and Musculoskeletal Modeling to Investigate Joint Loading in a Clinical Environment
Authors: Holder Jana , Trinler Ursula , Meurer Andrea , Stief Felix
Published: December 2020

The assessment of knee or hip joint loading by external joint moments is mainly used to draw conclusions on clinical decision making. However, the correlation between internal and external loads has not been systematically analyzed. This systematic review aims, therefore, to clarify the relationship between external and internal joint loading measures during gait. A systematic database search was performed to identify appropriate studies for inclusion. In total, 4,554 articles were identified, while 17 articles were finally included in data extraction. External joint loading parameters were calculated using the inverse dynamics approach and internal joint loading parameters by musculoskeletal modeling or instrumented prosthesis. It was found that the medial and total knee joint contact forces as well as hip joint contact forces in the first half of stance can be well predicted using external joint moments in the frontal plane, which is further improved by including the sagittal joint moment. Worse correlations were found for the peak in the second half of stance as well as for internal lateral knee joint contact forces. The estimation of external joint moments is useful for a general statement about the peak in the first half of stance or for the maximal loading. Nevertheless, when investigating diseases as valgus malalignment, the estimation of lateral knee joint contact forces is necessary for clinical decision making because external joint moments could not predict the lateral knee joint loading sufficient enough. Dependent on the clinical question, either estimating the external joint moments by inverse dynamics or internal joint contact forces by musculoskeletal modeling should be used.
Numerical Modeling of RC Columns and a Modified Steel Model Proposal for Elements With Plain Bars
Authors: Melo José , Varum Humberto , Rossetto Tiziana
Published: December 2020

The cyclic earthquake loads may accelerate the bond degradation and consequently lead to important bar slippage. The bond-slip mechanism is assumed as a common cause of damage or collapse of existing RC structures loaded by seismic leads. The RC structures designed and built before the implementation of the modern seismic codes and with plain reinforcing bars are particularly affected by the bond degradation. However, perfect bond is assumed in most of the numerical models. The numerical modeling results of two RC columns tested under cyclic lateral load are presented in this paper. One column is built with plain reinforcing bars and the other with deformed reinforcing bars and both have structural detailing typically adopted in pre-1970's structures. For each column, different software and modeling strategies to simulate the cyclic response were adopted. The frameworks OpenSees and SeismoStruct were used to develop the numerical models which were calibrated based on the experimental results. A simple modeling strategy was adopted in the OpenSees models to consider the bond-slip effects. A modified tri-linear steel material model is proposed and adopted to contemplate the slippage of plain reinforcing bars by reducing the steel Young modulus. The tri-linear steel model parameters were obtained empirically based on the experimental results.
Current Methods for Detecting Cell Membrane Transient Interactions
Authors: Bagheri Yousef , Ali Ahsan Ausaf , You Mingxu
Published: December 2020

Short-lived cell membrane complexes play a key role in regulating cell signaling and communication. Many of these complexes are formed based on low-affinity and transient interactions among various lipids and proteins. New techniques have emerged to study these previously overlooked membrane transient interactions. Exciting functions of these transient interactions have been discovered in cellular events such as immune signaling, host–pathogen interactions, and diseases such as cancer. In this review, we have summarized current experimental methods that allow us to detect and analyze short-lived cell membrane protein–protein, lipid–protein, and lipid–lipid interactions. These methods can provide useful information about the strengths, kinetics, and/or spatial patterns of membrane transient interactions. However, each method also has its own limitations. We hope this review can be used as a guideline to help the audience to choose proper approaches for studying membrane transient interactions in different membrane trafficking and cell signaling events.
Revealing the Disturbed Vaginal Micobiota Caused by Cervical Cancer Using High-Throughput Sequencing Technology
Authors: Xie Yupei , Feng Ying , Li Wenyu , Zhan Fuliang , Huang Genhua , Hu Hui , Xiong Yifei , Tan Buzhen , Chen Tingtao
Published: December 2020

Cervical cancer is the fourth most prevalent cancer type among all malignancies, so it is of great significance to find its actual pathogenesis mechanisms. In the present study, 90 women were enrolled, and high-throughput sequencing technology was firstly used to analyze the vaginal microbiota of healthy women (C group), cervical intraepithelial neoplasia patients (CIN group) and cervical cancer patients (CER group). Our results indicates that compared with C group, a higher HPV infection rate as well as increased Neutrophil ratio and tumor marker squamous cell carcinoma antigen (SCCA) were obtained, and a decrease in Lymphocyte ratio and Hemoglobin were also present. In addition, the cervical cancer showed a strong association with reduced probiotics Lactobacillus, increased pathogens Prevotella spp., Sneathia spp. and Pseudomonas spp. These results prove that the immunological changes generated by the cervical cancer and the vaginal microbiota can interact with each other. However, further study investigating the key bacteria for cervical cancer is still needed, which can be a clue for the diagnosis or treatment of cervical cancer.
Clinical Characteristics and Immune Responses of 137 Deceased Patients With COVID-19: A Retrospective Study
Authors: Cui Ning , Yan Rongdi , Qin Chunyuan , Zhao Jingming
Published: December 2020

Objective

This study aimed to evaluate the factors associated with death in patients with coronavirus disease 2019 by clarifying the clinical characteristics and immune responses.

Methods

The clinical characteristics and laboratory findings, including cytokine and lymphocyte subsets, were obtained from the electronic medical records of patients in Wuhan Tongji Hospital.

Results

This study included 836 patients with confirmed COVID-19. In total, 699 (83.6%) were cured and discharged, and 137 (16.4%) died. Our analysis revealed that age ≥ 65 years, male sex, malignancy, chronic obstructive pulmonary disease, dyspnea, dizziness, respiratory rate > 20 bpm, heart rate > 100 bpm, systolic blood pressure < 90 mmHg, neutrophils > 6.3×109/L, lymphopenia, thrombocytopenia, D-dimer ≥ 0.5 mg/L, lactate dehydrogenase > 250 U/L, aspartate aminotransferase > 40 U/L, total bilirubin > 26 μmol/L, albumin < 35 g/L, blood urea nitrogen > 9.5 mmol/L, estimated glomerular filtration rate < 90 ml/min/1.73, elevated cardiac troponin I, N-terminal pro-brain natriuretic peptide ≥ 900 pg/ml, C-reactive protein ≥ 25 mg/L, procalcitonin ≥ 0.05 ng/ml and ferritin > 400 μg/L were associated with death in patients with COVID-19. The multivariate logistic regression analysis revealed that an estimated glomerular filtration rate < 90 ml/min/1.73, elevated cardiac troponin I, C-reactive protein ≥ 25 mg/L and procalcitonin ≥ 0.05 ng/ml were predictive of mortality. Regarding immune responses, IL-2R, IL-6, IL-8, IL-10, and TNFα were remarkably higher in the deceased group at admission, and the levels of IL-2R, IL-6, IL-8, IL-10, and TNFα in the deceased group showed a rapid increase; the dynamics of these cytokines were highly consistent with disease deterioration. Lymphocyte subset analysis revealed that the deceased patients showed significant decreases in lymphocyte counts, especially helper T cells, suppressor T cells and NK cells.

Conclusions

This study identified that an estimated glomerular filtration rate < 90 ml/min/1.73, elevated cardiac troponin I, C-reactive protein ≥ 25 mg/L and procalcitonin ≥ 0.05 ng/ml were predictors of mortality in COVID-19 patients. Elevated cytokine levels and a continued increasing trend, including in IL-2R, IL-6, IL-8, IL-10 and TNFα, and a decrease in lymphocyte subsets, especially helper T cells, suppressor T cells and NK cells, were associated with a poor prognosis.
Cardioprotective Effect of Decorin in Type 2 Diabetes
Authors: Chen Fuqiong , Lai Jinsheng , Zhu Yanfang , He Mengying , Hou Huiying , Wang Jin , Chen Chen , Wang Dao Wen , Tang Jiarong
Published: December 2020

Cardiomyopathy is the leading cause of increased mortality in diabetes. In the present study, we investigated the effects of decorin (DCN) gene therapy on left ventricular function, cardiac inflammation and fibrosis in type 2 diabetes. Type 2 diabetes was induced in male Wistar rats by high fat diet (HFD, 60% of calories as fat) and STZ (20 mg/kg, intraperitoneal). Diabetic rats were divided into (n=6 for each group) the control group, the GFP-treated group and the DCN-treated group, received intravenous injection of saline solution, recombinant adeno-associated viral (rAAV)-GFP, and rAAV-DCN, respectively. We evaluated cardiac inflammation, fibrosis, left ventricular function at 6 months after gene delivery. Results turned out that rAAV-DCN treatment attenuated diabetic cardiomyopathy with improved LV function compared with control animals, which might be related to the reduced cardiac inflammation and fibrosis. These protective effects were associated with TGFβ1 pathway (ERK1/2 and smad-2) and NF-κB pathway, which may due to the decreased activation level of IGF-IR, increased expression of PKC-α and Hsp70. In conclusion, our results show that rAAV-mediated DCN therapy may be beneficial in the treatment of Diabetic Cardiomyopathy.
Interactions Between Maternal, Paternal, Developmental, and Immediate Environmental Effects on Anti-predator Behavior of the Snail Physa acuta
Authors: Tariel Juliette , Luquet Émilien , Plénet Sandrine
Published: December 2020

Transgenerational plasticity, which occurs when the environment experienced by parents changes the phenotype of offspring, is widespread in animal and plant species. Both maternal and paternal environments can underlie transgenerational plasticity, but experimental studies unraveling how their effects interact together and with the personal (both developmental and immediate) environments are still rare. Yet unraveling these interactions is fundamental to understanding how offspring integrate past and present environmental cues to produce adaptive phenotype. Using the hermaphroditic and freshwater snail Physa acuta, we tested how predator cues experienced by offspring, mothers and fathers interact to shape offspring anti-predator behavior. We raised a first generation of snails in the laboratory with or without chemical predator cues and realized full-factorial crosses to disentangle maternal and paternal cues. We then raised the second generation of snails with or without predator cues and assessed, when adults, their escape behavior in two immediate environments (with or without predator cues) and activity in the immediate environment without predator cues. We found that personal, maternal, and paternal predator cues interacted to shape offspring escape behavior and activity. Firstly, for escape behavior, snails integrated the cues from developmental and parental environments only when exposed to predator cues in their immediate environment, suggesting that personal immediate experience must corroborate the risky parental environment to reveal transgenerational plasticity. For activity, this same hypothesis helps explain why no clear pattern of transgenerational plasticity was revealed, as activity was only measured without predator cues in the immediate environment. Secondly, a single maternal exposure to predator cues decreased offspring escape behavior while a single paternal exposure had no effect, surprisingly demonstrating sex-specific transgenerational plasticity for a simultaneous hermaphroditic species. Thirdly, when both mother and father were exposed, paternal cues were integrated by offspring according to their own developmental environment. The paternal exposure then mitigated the reduction in escape behavior due to the maternal exposure only when offspring developed in control condition. Overall, our study highlighted complex patterns of sex-specific transgenerational plasticity resulting from non-additive interactions between parental, developmental and immediate experiences.
Species Delimitation of the Eisenia nordenskioldi Complex (Oligochaeta, Lumbricidae) Using Transcriptomic Data
Authors: Shekhovtsov Sergei V. , Shipova Aleksandra A. , Poluboyarova Tatiana V. , Vasiliev Gennady V. , Golovanova Elena V. , Geraskina Anna P. , Bulakhova Nina A. , Szederjesi Tímea , Peltek Sergei E.
Published: December 2020

Eisenia nordenskioldi (Eisen, 1879) is the only autochthonous Siberian earthworm with a large distribution that ranges from tundra to steppe and broadleaved forests. This species has a very high morphological, ecological, karyological, and genetic diversity, so it was proposed that E. nordenskioldi should be split into several species. However, the phylogeny of the complex was unclear due to the low resolution of the methods used and the high diversity that should have been taken into account. We investigated this question by (1) studying the diversity of the COI gene of E. nordenskioldi throughout its range and (2) sequencing transcriptomes of different genetic lineages to infer its phylogeny. We found that E. nordenskioldi is monophyletic and is split into two clades. The first one includes the pigmented genetic lineages widespread in the northern and western parts of the distribution, and the second one originating from the southern and southeastern part of the species' range and representing both pigmented and non-pigmented forms. We propose to split the E. nordenskioldi complex into two species, E. nordenskioldi and Eisenia sp. 1 (aff. E. nordenskioldi), corresponding to these two clades. The currently recognized non-pigmented subspecies E. n. pallida will be abolished as a polyphyletic and thus a non-natural taxon, while Eisenia sp. 1 will be expanded to include several lineages earlier recognized as E. n. nordenskioldi and E. n. pallida.
Chromatin Structure and Function in Mosquitoes
Authors: Lezcano Óscar M. , Sánchez-Polo Miriam , Ruiz José L. , Gómez-Díaz Elena
Published: December 2020

The principles and function of chromatin and nuclear architecture have been extensively studied in model organisms, such as Drosophila melanogaster. However, little is known about the role of these epigenetic processes in transcriptional regulation in other insects including mosquitoes, which are major disease vectors and a worldwide threat for human health. Some of these life-threatening diseases are malaria, which is caused by protozoan parasites of the genus Plasmodium and transmitted by Anopheles mosquitoes; dengue fever, which is caused by an arbovirus mainly transmitted by Aedes aegypti; and West Nile fever, which is caused by an arbovirus transmitted by Culex spp. In this contribution, we review what is known about chromatin-associated mechanisms and the 3D genome structure in various mosquito vectors, including Anopheles, Aedes, and Culex spp. We also discuss the similarities between epigenetic mechanisms in mosquitoes and the model organism Drosophila melanogaster, and advocate that the field could benefit from the cross-application of state-of-the-art functional genomic technologies that are well-developed in the fruit fly. Uncovering the mosquito regulatory genome can lead to the discovery of unique regulatory networks associated with the parasitic life-style of these insects. It is also critical to understand the molecular interactions between the vectors and the pathogens that they transmit, which could hold the key to major breakthroughs on the fight against mosquito-borne diseases. Finally, it is clear that epigenetic mechanisms controlling mosquito environmental plasticity and evolvability are also of utmost importance, particularly in the current context of globalization and climate change.
NOD2/RIG-I Activating Inarigivir Adjuvant Enhances the Efficacy of BCG Vaccine Against Tuberculosis in Mice
Authors: Khan Arshad , Singh Vipul K. , Mishra Abhishek , Soudani Emily , Bakhru Pearl , Singh Christopher R. , Zhang Dekai , Canaday David H. , Sheri Anjaneyulu , Padmanabhan Seetharamaiyer , Challa Sreerupa , Iyer Radhakrishnan P. , Jagannath Chinnaswamy
Published: December 2020

Tuberculosis (TB) caused by Mycobacterium tuberculosis (MTB) kills about 1.5 million people each year and the widely used Bacille Calmette-Guérin (BCG) vaccine provides a partial protection against TB in children and adults. Because BCG vaccine evades lysosomal fusion in antigen presenting cells (APCs), leading to an inefficient production of peptides and antigen presentation required to activate CD4 T cells, we sought to boost its efficacy using novel agonists of RIG-I and NOD2 as adjuvants. We recently reported that the dinucleotide SB 9200 (Inarigivir) derived from our small molecule nucleic acid hybrid (SMNH)® platform, activated RIG-I and NOD2 receptors and exhibited a broad-spectrum antiviral activity against hepatitis B and C, Norovirus, RSV, influenza and parainfluenza. Inarigivir increased the ability of BCG-infected mouse APCs to secrete elevated levels of IL-12, TNF-α, and IFN-β, and Caspase-1 dependent IL-1β cytokine. Inarigivir also increased the ability of macrophages to kill MTB in a Caspase-1-, and autophagy-dependent manner. Furthermore, Inarigivir led to a Capsase-1 and NOD2- dependent increase in the ability of BCG-infected APCs to present an Ag85B-p25 epitope to CD4 T cells in vitro. Consistent with an increase in immunogenicity of adjuvant treated APCs, the Inarigivir-BCG vaccine combination induced robust protection against tuberculosis in a mouse model of MTB infection, decreasing the lung burden of MTB by 1-log10 more than that afforded by BCG vaccine alone. The Inarigivir-BCG combination was also more efficacious than a muramyl-dipeptide-BCG vaccine combination against tuberculosis in mice, generating better memory T cell responses supporting its novel adjuvant potential for the BCG vaccine.
Key Metabolic Pathways in MSC-Mediated Immunomodulation: Implications for the Prophylaxis and Treatment of Graft Versus Host Disease
Authors: Burnham Andre J. , Foppiani Elisabetta Manuela , Horwitz Edwin M.
Published: December 2020

Mesenchymal stromal cells (MSCs) are spindle-shaped, plastic-adherent cells in vitro with potent immunosuppressive activity both in vitro and in vivo. MSCs have been employed as a cellular immunotherapy in diverse preclinical models and clinical trials, but most commonly as agents for the prophylaxis or therapy of graft versus host disease after hematopoietic cell transplantation. In addition to the oft studied secreted cytokines, several metabolic pathways intrinsic to MSCs, notably indoleamine 2,3-dioxygenase, prostaglandin E2, hypoxia-inducible factor 1 α, heme oxygenase-1, as well as energy-generating metabolism, have been shown to play roles in the immunomodulatory activity of MSCs. In this review, we discuss these key metabolic pathways in MSCs which have been reported to contribute to MSC therapeutic effects in the setting of hematopoietic cell transplantation and graft versus host disease. Understanding the contribution of MSC metabolism to immunomodulatory activity may substantially inform the development of future clinical applications of MSCs.
Management of Cervical Cancer in Pregnant Women: A Multi-Center Retrospective Study in China
Authors: Li Mingzhu , Zhao Yun , Qie Mingrong , Zhang Youzhong , Li Longyu , Lin Bei , Guo Ruixia , You Zhixue , An Ruifang , Liu Jun , Zhang Zhijun , Bi Hui , Hong Ying , Chang Shufang , He Guoli , Hua Keqin , Zhou Qi , Liao Qinping , Wang Yue , Wang Jianliu , Li Xiaoping , Wei Lihui
Published: December 2020

Background: This retrospective multi-center study aimed to describe the epidemiological characteristics, clinical features, and management of patients with cervical cancer in pregnancy (CCIP) and evaluate maternal and infant outcomes.

Methods: The data of patients with CCIP were retrospectively collected from those diagnosed and treated in 17 hospitals in 12 provinces in China between January 2009 and November 2017. The information retrieved included patients' age, clinical features of the tumor, medical management (during pregnancy or postpartum), obstetrical indicators (i.e., gestational age at diagnosis, delivery mode, and birth weight), and maternal and neonatal outcomes. Survival analyses were performed using Kaplan–Meier survival curves and log-rank tests that estimated the overall survival of patients.

Results: One-hundred and five women diagnosed with CCIP (median age = 35 years) were identified from ~45,600 cervical cancer patients (0.23%) and 525,000 pregnant women (0.020%). The median gestational age at cancer diagnosis was 20.0 weeks. The clinical-stage of 93.3% of the patients with CCIP was IB1, 81.9% visited the clinic because of vaginal bleeding during pregnancy, and 72.4% had not been screened for cervical cancer in more than 5 years. To analyze cancer treatments during pregnancy, patients were grouped into two groups, termination of pregnancy (TOP, n = 67) and continuation of pregnancy (COP, n = 38). Analyses suggested that the TOP group was more likely to be diagnosed at an earlier gestational stage than the COP group (14.8 vs. 30.8 weeks, p < 0.001). The unadjusted hazard ratio for the COP group's overall survival was 1.063 times that of the TOP group (95% confidence interval = 0.24, 4.71). There were no significant differences between the TOP and COP groups in maternal survival (p = 0.964). Thirty-three of the infants of patients with CCIP were healthy at the end of the follow-up period, with a median age of 18 ± 2.8 months.

Conclusions: Most patients with CCIP had not been screened for cervical cancer in over 5 years. The oncologic outcomes of the TOP and COP groups were similar. A platinum-based neoadjuvant chemotherapy regimen could be a favorable choice for the management of CCIP during the second and third trimesters of pregnancy.
Bone Marrow Mesenchymal Stem Cell-Derived Exosomal miR-25 Regulates the Ubiquitination and Degradation of Runx2 by SMURF1 to Promote Fracture Healing in Mice
Authors: Jiang Yikun , Zhang Jun , Li Zhengwei , Jia Guoliang
Published: December 2020

Recent evidence has demonstrated that mesenchymal stem cells (MSCs) can release a large number of functionally specific microRNA (miRNA) microvesicles that play a role in promoting osteogenic differentiation, but the specific mechanism is not yet clear. Under such context, this study aims to elucidate the mechanism of bone marrow mesenchymal stem cell-derived exosomes (BMSC-Exo) promoting fracture healing in mice. We isolated and identified the BMSC-Exo. Bioinformatics analysis predicted high expression of miRNA in exosomes and verified the transfer of miR-25 in exosomes by immunofluorescence. Targeting relationship between miR-25 and Smad ubiquitination regulatory factor-1 (SMURF1) was predicted and verified by dual-luciferase reporter gene assay. Immunoprecipitation and protein stability assays were used to detect Runt-related transcription factor 2 (Runx2) ubiquitination and the effect of SMURF1 on Runx2 ubiquitination, respectively. The effect of miR-25 in BMSC-Exo on fracture healing in mice was assessed using X-ray imaging. alkaline phosphatase, alizarin red staining, EdU, CCK-8, and Transwell were used to evaluate the effects of exosomes transferred miR-25 on osteogenic differentiation, proliferation, and migration of osteoblasts. Bioinformatics analysis predicted that miR-25 expression in exosomes increased significantly. Moreover, the targeted regulation of SMURF1 by miR-25 was verified. SMURF1 inhibited Runx2 protein expression by promoting ubiquitination degradation of Runx2. Notably, miR-25 secreted by BMSC-Exo can accelerate osteogenic differentiation, proliferation, and migration of osteoblasts through SMURF1/Runx2 axis. Our results demonstrate that miR-25 in BMSC-Exo regulates the ubiquitination degradation of Runx2 by SMURF1 to promote fracture healing in mice.
An Educational Bioinformatics Project to Improve Genome Annotation
Authors: Amatore Zoie , Gunn Susan , Harris Laura K.
Published: December 2020

Scientific advancement is hindered without proper genome annotation because biologists lack a complete understanding of cellular protein functions. In bacterial cells, hypothetical proteins (HPs) are open reading frames with unknown functions. HPs result from either an outdated database or insufficient experimental evidence (i.e., indeterminate annotation). While automated annotation reviews help keep genome annotation up to date, often manual reviews are needed to verify proper annotation. Students can provide the manual review necessary to improve genome annotation. This paper outlines an innovative classroom project that determines if HPs have outdated or indeterminate annotation. The Hypothetical Protein Characterization Project uses multiple well-documented, freely available, web-based, bioinformatics resources that analyze an amino acid sequence to (1) detect sequence similarities to other proteins, (2) identify domains, (3) predict tertiary structure including active site characterization and potential binding ligands, and (4) determine cellular location. Enough evidence can be generated from these analyses to support re-annotation of HPs or prioritize HPs for experimental examinations such as structural determination via X-ray crystallography. Additionally, this paper details several approaches for selecting HPs to characterize using the Hypothetical Protein Characterization Project. These approaches include student- and instructor-directed random selection, selection using differential gene expression from mRNA expression data, and selection based on phylogenetic relations. This paper also provides additional resources to support instructional use of the Hypothetical Protein Characterization Project, such as example assignment instructions with grading rubrics, links to training videos in YouTube, and several step-by-step example projects to demonstrate and interpret the range of achievable results that students might encounter. Educational use of the Hypothetical Protein Characterization Project provides students with an opportunity to learn and apply knowledge of bioinformatic programs to address scientific questions. The project is highly customizable in that HP selection and analysis can be specifically formulated based on the scope and purpose of each student’s investigations. Programs used for HP analysis can be easily adapted to course learning objectives. The project can be used in both online and in-seat instruction for a wide variety of undergraduate and graduate classes as well as undergraduate capstone, honor’s, and experiential learning projects.
Transcriptome Profiling Provides Insights Into Potential Antagonistic Mechanisms Involved in Chaetomium globosum Against Bipolaris sorokiniana
Authors: Darshan K. , Aggarwal Rashmi , Bashyal Bishnu Maya , Singh Jagmohan , Shanmugam V. , Gurjar Malkhan S. , Solanke Amolkumar U.
Published: December 2020

Chaetomium globosum Kunze is recognized as a potential biocontrol fungus against spot blotch of wheat caused by Bipolaris sorokiniana. Its molecular mechanism of biocontrol activity and the biosynthetic pathways involved have not been yet elucidated. Here, global transcriptome profiling of C. globosum strain Cg2 during interaction with B. sorokiniana isolate BS112 using RNA-seq was performed in order to gain insights into the potential mechanisms of antagonism. The Illumina HiSeq platform (2 × 150 bp) yielded an average of 20–22 million reads with 50–58% GC. De novo assembly generated 45,582 transcripts with 27,957 unigenes. Transcriptome analysis displayed distinct expression profiles in the interaction (Cg2–BS112), out of which 6,109 unique differentially expressed genes were present. The predominant transcripts classified as genes involved in “catalytic activity” constituted 45.06%, of which 10.02% were associated with “hydrolytic activity” (GO:0008152), and similarly, in the biological process, 29.18% of transcripts were involved in “metabolic activity” (GO:0004096 and GO:0006979). Heat map and cluster categorization suggested an increase in the expression levels of genes encoding secondary metabolites like polyketide synthase (GO:0009058), S-hydroxymethyl glutathione dehydrogenase (GO:0006069), terpene cyclase (EC 4.2.3.-), aminotran_1_2 domain-containing protein (GO:0009058), and other hydrolytic CAZYmes such as the glycosyl hydrolase (GH) family (GH 13, GH 2, GH 31, and GH 81; GO:0005975), cellulase domain-containing protein, chitinases, β-1, 3-glucanases (GO:0004565), glucan endo-1,3-beta-glucanase (GO:0052861), and proteases (GO:0004177). The obtained RNA-seq data were validated by RT-qPCR using 20 randomly chosen genes, showing consistency with the RNA-seq results. The present work is worldwide the first effort to unravel the biocontrol mechanism of C. globosum against B. sorokiniana. It generated a novel dataset for further studies and facilitated improvement of the gene annotation models in the C. globosum draft genome.
Protein Residues and a Novel Motif Involved in the Cellular Localization of CheZ in Azorhizobium caulinodans ORS571
Authors: Liu Xiaolin , Liu Yanan , Johnson Kevin Scot , Dong Xiaoyan , Xie Zhihong
Published: December 2020

Chemotaxis is essential for the competitiveness of motile bacteria in complex and harsh environments. The localization of chemotactic proteins in the cell is critical for coordinating a maximal response to chemotactic signals. One chemotaxis protein with a well-defined subcellular localization is the phosphatase CheZ. CheZ localizes to cell poles by binding with CheA inEscherichia coliand other enteric bacteria, or binding with a poorly understood protein called ChePep in epsilon-Proteobacteria. In alpha-Proteobacteria, CheZ lacks CheA-binding sites, and its cellular localization remains unknown. We therefore determined the localization of CheZ in the alpha-Proteobacteria Azorhizobium caulinodansORS571.A. caulinodansCheZ, also termed as CheZAC,was found to be located to cell poles independently of CheA, and we suspect that either the N-terminal helix or the four-helix bundle of CheZACis sufficient to locate to cell poles. We also found a novel motif, AXXFQ, which is adjacent to the phosphatase active motif DXXXQ, which effects the monopolar localization of CheZAC. This novel motif consisting of AXXFQ is conserved in CheZ and widely distributed amongProteobacteria. Finally, we found that the substitution of phosphatase active site affects the polar localization of CheZAC. In total, this work characterized the localization pattern of CheZ containing a novel motif, and we mapped the regions of CheZACthat are critical for its polar localization.
Discovery of Novel Biosynthetic Gene Cluster Diversity From a Soil Metagenomic Library
Authors: Santana-Pereira Alinne L. R. , Sandoval-Powers Megan , Monsma Scott , Zhou Jinglie , Santos Scott R. , Mead David A. , Liles Mark R.
Published: December 2020

Soil microorganisms historically have been a rich resource for natural product discovery, yet the majority of these microbes remain uncultivated and their biosynthetic capacity is left underexplored. To identify the biosynthetic potential of soil microorganisms using a culture-independent approach, we constructed a large-insert metagenomic library inEscherichia colifrom a topsoil sampled from the Cullars Rotation (Auburn, AL, United States), a long-term crop rotation experiment. Library clones were screened for biosynthetic gene clusters (BGCs) using either PCR or a NGS (next generation sequencing) multiplexed pooling strategy, coupled with bioinformatic analysis to identify contigs associated with each metagenomic clone. A total of 1,015 BGCs were detected from 19,200 clones, identifying 223 clones (1.2%) that carry a polyketide synthase (PKS) and/or a non-ribosomal peptide synthetase (NRPS) cluster, a dramatically improved hit rate compared to PCR screening that targeted type I polyketide ketosynthase (KS) domains. The NRPS and PKS clusters identified by NGS were distinct from known BGCs in the MIBiG database or those PKS clusters identified by PCR. Likewise, 16S rRNA gene sequences obtained by NGS of the library included many representatives that were not recovered by PCR, in concordance with the same bias observed in KS amplicon screening. This study provides novel resources for natural product discovery and circumvents amplification bias to allow annotation of a soil metagenomic library for a more complete picture of its functional and phylogenetic diversity.
Brain Temperature Alters Contributions of Excitatory and Inhibitory Inputs to Evoked Field Potentials in the Rat Frontal Cortex
Authors: Gotoh Mizuho , Nagasaka Kazuaki , Nakata Mariko , Takashima Ichiro , Yamamoto Shinya
Published: December 2020

Changes in brain temperature have been reported to affect various brain functions. However, little is known about the effects of temperature on the neural activity at the network level, where multiple inputs are integrated. In this study, we recorded cortical evoked potentials while altering the local brain temperature in anesthetized rats. We delivered electrical stimulations to the midbrain dopamine area and measured the evoked potentials in the frontal cortex, the temperature of which was locally altered using a thermal control device. We focused on the maximum negative peaks, which was presumed to result mainly from polysynaptic responses, to examine the effect of local temperature on network activity. We showed that focal cortical cooling increased the amplitude of evoked potentials (negative correlation, >17°C); further cooling decreased their amplitude. This relationship would be graphically represented as an inverted-U-shaped curve. The pharmacological blockade of GABAergic inhibitory inputs eliminated the negative correlation (>17°C) and even showed a positive correlation when the concentration of GABAA receptor antagonist was sufficiently high. Blocking the glutamatergic excitatory inputs decreased the amplitude but did not cause such inversion. Our results suggest that the negative correlation between the amplitude of evoked potentials and the near-physiological local temperature is caused by the alteration of the balance of contribution between excitatory and inhibitory inputs to the evoked potentials, possibly due to higher temperature sensitivity of inhibitory inputs.
Viruses Causing Aseptic Meningitis: A Tertiary Medical Center Experience With a Multiplex PCR Assay
Authors: Aldriweesh Mohammed A. , Shafaay Edi A. , Alwatban Saud M. , Alkethami Obeid M. , Aljuraisi Faisal N. , Bosaeed Mohammad , Alharbi Naif Khalaf
Published: December 2020

Background: Central nervous system (CNS) infection is associated with high rates of morbidity and mortality, and despite advancements in molecular testing, aseptic meningitis remains challenging to diagnose. Aseptic meningitis cases are often underreported worldwide, which impacts the quality of patient care. Therefore, we aimed to assess the results of BioFire® FilmArray® meningitis/encephalitis (ME) PCR panel, clinical characteristics, and etiologies of aseptic meningitis patients.

Methods: From January 2018 to January 2020, all pediatric and adult patients in a large tertiary medical center who underwent lumbar puncture and cerebrospinal fluid (CSF) testing by a ME multiplex PCR panel and who fit the aseptic meningitis definition were retrospectively reviewed.

Results: Data were reviewed from 1,607 patients; 240 met the inclusion criteria (54.6% males; 68.8% <4 years of age). The rate of detected viral causes of aseptic meningitis was 40.4%; therefore, 59.6% of the patients remained with unidentified etiology. Among the identified viral meningitis, enterovirus and human herpesvirus 6 (HHV-6) were the most common (25 and 7.9%, respectively). The median length of hospital stay was 6 days, and it was longer in patients with unidentifiable aseptic meningitis (p < 0.0001).

Conclusion: Aseptic meningitis is common among suspected meningitis patients, but most cases remained of unknown etiology. The most common identified viruses were enterovirus followed by HHV-6, and there is predominance in males and the pediatric age group. These results highlight that further research is needed to identify other etiologies and possible additional viral pathogens for aseptic meningitis in the current diagnostic methods.