bluuebunny/crossref_metadata_2025 huggingface dataset:
Lentinan combined with cisplatin for the treatment of non-small cell lung cancer
Authors: Zhao Chenchen , Yan Haifeng , Pang Wentai , Wu Tong , Kong Xianbin , Li Xiaojiang , Liu Honggen , Zhao Linlin , Liang Feng , Jia Yingjie
Published: March 2021

Background:

A growing number of studies suggest that lentinan combined with cisplatin thoracic injection for the treatment of lung cancer is an effective combination of traditional Chinese and Western medicine, which has a continuous and beneficial effect on eliminating clinical symptoms and improving cachexia in lung cancer patients. However, whether this treatment is effective and safe for lung cancer patients or not, evidence supporting the effectiveness and safety of this treatment is still incomplete. Besides, there is lack of systematic review to assess the detailed situation (including risk of bias and methodology) of current related clinical studies.

Objective:

This study aimed to evaluate the effectiveness and safety of lentinan combined with cisplatin thoracic injection in the treatment of lung cancer.

Methods:

The major databases (Embase, PubMed, the Cochrane Library, China National Knowledge Infrastructure, Chinese Scientific Journals Database [VIP] Database, Chinese Biomedical Literature Service System [SinoMed], and Wanfang Database) were searched from inception to March 1, 2020. Randomized controlled trials (RCTs) of lentinan combined with cisplatin chest injection on patients with non-small cell lung cancer (NSCLC) were identified. Two assessors reviewed each trial independently. The methodological quality of the eligible studies was evaluated according to the Cochrane Collaboration's tool for assessing risk of bias. Both the data extraction and the literature quality screening evaluation were conducted independently by 2 researchers.

Results:

Totally 17 clinical RCTs were included in this study, involving 1390 patients. Meta-analysis results showed that the clinical efficacy (risk ratio [RR] = 1.34, 95% confidence interval [CI] 1.21–1.48), effective subgroup analysis (RR = 1.51, 95% CI 1.3–1.77), and quality of life (RR = 1.48, 95% CI 1.27–1.72), the differences are statistically significant. In terms of adverse reactions, mainly related to gastrointestinal reactions and bone marrow suppression, the incidence and degree of adverse reactions of lentinan combined with cisplatin thoracic injection group were lower than those of cisplatin thoracic injection group alone.

Conclusions:

The current evidence prompted that Lentinan combined with cisplatin in thoracic injection might benefit patients with NSCLC on a certain extent; this systematic review revealed some definite conclusions about the application of Lentinan combined with cisplatin in thoracic injection for NSCLC. Due to the low methodological quality, high risk of bias, and inadequate reporting on clinical data, these results still require verification by a large number of well designed, heterogeneous RCT studies. More rigorous, multicenter, sufficient-sample, and double-blind RCTs are warranted.
The risk assessment model of fracture nonunion after intramedullary nailing for subtrochanteric femur fracture
Authors: Wang ZhengHao , Li KaiNan , Gu ZuChao , Fan HaiQuan , Li HaiBo
Published: March 2021

To investigate the influencing factors of fracture nonunion after intramedullary nailing for subtrochanteric fractures and to construct a risk assessment model.

Based on the multicenter retrospective analysis of 251 patients, all patients were divided into modeling group and verification group. In the modeling group, postoperative fracture nonunion rate, general data, fracture-related factors, surgical reduction-related factors, mechanical and biological factors were calculated, and the influencing factors of fracture nonunion were screened by univariate analysis. Logistic regression model was used for multifactor analysis to construct the risk assessment model. Based on the logistic regression model, the risk prediction model was constructed by drawing the Nomogram diagram. Through the verification group, the influencing factors were evaluated again, and the differentiation and calibration of the model were evaluated. The calibration degree was evaluated by Hosmer-Lemeshow test, goodness of fit test, and calibration curve. The discriminant degree was evaluated by the receiver operating characteristic curve.

Fracture nonunion occurred in 34 of 149 patients in the modeling group. Among the 14 potential influencing factors, univariate analysis and logistic regression analysis showed that postoperative hip varus, intramedullary nail fixation failure, and reduction of fracture with large incision were the risk factors of fracture nonunion. The medial cortex fracture was seen reduced on X-Ray was a protective factor for fracture nonunion, and a regression equation was established. Based on the logistic regression model, the Nomogram diagram is drawn. Twenty-four cases of fracture nonunion occurred in the verification group. The area under the receiver operating characteristic curve was area under curve =0.883 > 0.7, indicating that there was a moderate differentiation to evaluate the occurrence of fracture nonunion after operation. The goodness of fit test: the Hosmers-Lemeshow test (X
2 = 2.921, P = .712 > .05) showed that the model had a good calibration.

After intramedullary nailing of subtrochanteric fracture, hip varus, failure of intramedullary nail fixation and wide surgical dissection are the risk factors of fracture nonunion, and the postoperative reduction of medial cortex fracture is protective factor.

National key research and development projects: 2016YFC0105806
The Relationship Between Pain and Quality of Life Among Adults With Knee Osteoarthritis
Authors: Park Youngrye , Park Sunah , Lee Mi Young
Published: March 2021

Pain, lower extremity functional status, and depression have been associated with quality of life among patients with knee osteoarthritis (KOA). However, little is known about the manner in which these factors influence quality of life. The present study examined the mediating effects of lower extremity functional status and depression on the relationship between pain and quality of life among adults with KOA. A descriptive cross-sectional study was conducted among 135 adults with KOA who visited an orthopaedic outpatient clinic in Korea. The participants responded to the Numeric Rating Scale, Lower Extremity Functional Scale, Beck Depression Inventory, and the Korean version of the World Health Organization Quality of Life-BREF. Mediation analysis was conducted using the PROCESS macro for SPSS. The serial mediating effects of lower extremity functional status and depression on the relationship between pain and quality of life were significant. These findings suggest that the assessment and management of lower extremity functional status and depression are important means by which the quality of life of adults with painful KOA can be improved.
Evaluating Tourniquet Efficacy Comparing Exsanguination Techniques for the Upper Extremity
Authors: Goodwin Tyler M. , Davies James P. , Jackson J. Benjamin , Millikin Mary , Walsh John J.
Published: March 2021

Tourniquets have been used in the medical setting for centuries and have become the gold standard when preparing patients for surgery, particularly in orthopaedic surgery. Upper extremity tourniquet use improves intraoperative visibility and identification of anatomy. It also decreases blood loss intraoperatively and improves the safety of orthopaedic procedures. Despite the widespread use of tourniquets and differing methods of limb exsanguination, little research has been done quantifying its efficacy. The purpose of this study was to compare gravity exsanguination to Esmarch exsanguination of the upper extremity prior to tourniquet inflation in a large patient sample. A plethysmographic method based on water displacement served as a surrogate for the blood volume exsanguinated. Control measurements of water displacement were obtained from both upper extremities without tourniquet inflation. Water displacement was then measured with both gravity and Esmarch exsanguination techniques. Gender, handedness, height, weight, body mass index, and age were recorded for volunteers and used as covariates. Change in mean water displacement from control (un-exsanguinated) arm and gravity alone measurement was 37.2 ml. Change in mean water displacement between control arm and mean Esmarch measurement was 56.3 ml. Exsanguination using Esmarch compared to gravity alone resulted in a 51.2% increase in blood removal. Only age had a significant interaction effect for the Esmarch method. Analysis revealed that age accounted for 21.4% of all variance in blood exsanguinated using the Esmarch method when compared to the control group. The Esmarch technique was more efficacious for all demographics measured, but most efficacious in subjects who were older than 40 years. This data reaffirms that gravity exsanguination is more efficacious than no tourniquet use at all, and that the Esmarch technique is more efficacious than gravity. To our knowledge, this study is the most robust of its kind to critically and objectively compare upper extremity exsanguination methods and overall tourniquet use by age and supports the common practice of Esmarch exsanguination in orthopaedic extremity surgery.
Preconception, Antepartum, and Peripartum Care for the Woman With a Spinal Cord Injury: A Review of the Literature
Authors: Wendel Michael P. , Whittington Julie R. , Pagan Megan E. , Whitcombe Dayna D. , Pates Jason A. , McCarthy Richard E. , Magann Everett F.
Published: March 2021

Importance

Spinal cord injury (SCI) may result in temporary or permanent loss of sensory, motor, and autonomic function, presenting unique medical and psychosocial challenges in women during their childbearing years.

Objective

The aim of this study was to review the literature and describe the spectrum of pregnancy considerations, complications, and evidence-based obstetric practices in women with SCI.

Evidence Acquisition

A literature search was undertaken using the search engines of PubMed and Web of Science using the terms “spinal cord injury” or “spinal cord complications” and “pregnancy outcomes” or “pregnancy complications.” The search was limited to the English language, and there was no restriction on the years searched.

Results

The search identified 174 abstracts, 50 of which are the basis for this review. Pregnancy in women who have experienced an SCI requires a multidisciplinary approach. Common complications during pregnancy include recurrent urinary tract infection, upper respiratory tract infection, hypertension, venous thromboembolism, and autonomic dysreflexia (AD), which is a potentially life-threatening complication. Obstetricians should avoid potential triggers and be familiar with acute management of AD. Postpartum complications include difficulty initiating and maintaining breastfeeding and increased risk of postpartum depression and other mental health issues.

Conclusions and Relevance

Obstetricians caring for women with an SCI must be familiar with the unique challenges and complications that may occur during pregnancy and puerperium.

Relevance Statement

An evidence-based literature review of the care of pregnant women with spinal cord injury.

Target Audience

Obstetricians, Obstetric Anesthesiologists

Learning Objectives

After completing this activity, the learner should be better able to evaluate published literature regarding SCI in pregnant patients; explain the unique complications that can occur in these patients; identify potential risk factors and management strategies of AD; and devise treatment and recommendations for optimal patient outcomes.
Spinal Muscular Atrophy: Inheritance, Screening, and Counseling for the Obstetric Provider
Authors: Hopkins Maeve K. , Dugoff Lorraine , Kuller Jeffrey A.
Published: March 2021

Importance

Spinal muscular atrophy (SMA) confers significant risk of neonatal and infant morbidity and mortality. Screening women during or before pregnancy for carrier status of SMA presents an opportunity to identify pregnancies at risk for this potentially devastating condition.

Objective

The objective of this review is to describe the different forms of SMA and their inheritance. In addition, this review guides obstetric providers in interpreting results of carrier screening.

Evidence Acquisition

A MEDLINE search of “prenatal genetic testing,” “spinal muscular atrophy,” and “inheritance of spinal muscular atrophy” in the review was performed.

Results

The evidence cited in this review includes 4 medical society committee opinions and 14 additional peer-reviewed journal articles that were original research or expert opinion summaries.

Conclusions and Relevance

Spinal muscular atrophy is a severe, heterogeneous neurodegenerative disorder. The American College of Obstetricians and Gynecologists recommends that obstetricians offer carrier screening for SMA to all pregnant women. Given the different types and inheritance of SMA, understanding of the disease and interpreting carrier screening results is of paramount importance to the prenatal care provider.

Target Audience

Obstetrician-gynecologist, family medicine, or other practitioner(s) providing prenatal care.

Learning Objectives

After participating in this activity, physicians should be better able to identify indications for carrier testing for SMA; compare types of SMA based on age of onset and phenotype; interpret results of SMA carrier screening; counsel patients regarding residual risk after negative SMA carrier screening; and refer patients with positive SMA carrier screening.
Maternal Plasma Genome-Wide Cell-Free DNA Can Detect Fetal Aneuploidy in Early and Recurrent Pregnancy Loss and Can Be Used to Direct Further Workup
Authors: Yaron Yuval , Pauta Montse , Badenas Celia , Soler Anna , Borobio Virginia , Illanes Carmen , Paz-y-Miño Fernanda , Martinez-Portilla Raigam , Borrell Antoni
Published: March 2021

(Abstracted from Hum Reprod 2020;35(5):1222–1229)

Traditional diagnostic investigations of recurrent pregnancy loss (RPL) identify an etiology in fewer than half of cases. The single most common etiology is random chromosomal anomalies, and it has been established that 50% to 70% of early pregnancy losses (EPLs) are caused by numeric chromosomal anomalies.
Endometrial Cancer
Authors: Lu Karen H. , Broaddus Russell R.
Published: March 2021

(Abstracted from N Engl J Med 2020;383:2053–2064)

Incidence rates and associated mortality of endometrial cancer continue to rise in the United States, including a sustained rise in cases among women younger than 50 years. This trend is partially driven by the rising rates of obesity, one of the most important risk factors for this disease.
Vaginal Estrogen Therapy Is Associated With Increased Lactobacillus in the Urine of Postmenopausal Women With Overactive Bladder Symptoms
Authors: Thomas-White Krystal , Taege Susanne , Limeira Roberto , Brincat Cynthia , Joyce Cara , Hilt Evann E. , Mac-Daniel Laura , Radek Katherine A. , Brubaker Linda , Mueller Elizabeth R. , Wolfe Alan J.
Published: March 2021

(Abstracted from Am J Obstet Gynecol 2020;223:727.e1–727.e11)

Menopause is primarily the consequence of estrogen loss. It is associated with a wide variety of uncomfortable symptoms that affect the quality of life of postmenopausal women.
Associations Between Elevated Systolic Blood Pressure and Outcomes in Critically Ill Patients: A Retrospective Cohort Study and Propensity Analysis
Authors: Hu An-Min , Hai Chao , Wang Hai-Bo , Zhang Zhongheng , Sun Ling-Bin , Zhang Zhong-Jun , Li Hui-Ping
Published: March 2021

Introduction:

Studies have shown nonlinear relationships between systolic blood pressure (SBP) and outcomes, with increased risk observed at both low and high blood pressure levels. However, the relationships between cumulative times at different SBP levels and outcomes in critically ill patients remain unclear. We hypothesized that an appropriate SBP level is associated with a decrease in adverse outcomes after intensive care unit (ICU) admission.

Methods:

This study was a retrospective analysis of data from the Medical Information Mart for Intensive Care (MIMIC) III database, which includes more than 1,000,000 SBP records from 12,820 patients. Associations of cumulative times at four SBP ranges (<100 mm Hg, 100–120 mm Hg, 120–140 mm Hg, and ≥140 mm Hg) with mortality (12-, 3-, 1-month mortality and in-hospital mortality) were evaluated. Restricted cubic splines and multivariable Cox regression models were employed to assess associations between mortality and cumulative times at SBP levels (4 levels: <2, 2–12, 12–36, and ≥36 h) over 72 h of ICU admission. Additionally, 120 mm Hg to 140 mm Hg was subdivided into <12 h (Group L) and ≥12 h (Group M) subsets and subjected to propensity-score matching and subgroup analyses.

Results:

At 120 mm Hg to 140 mm Hg, level-4 SBP was associated with lower adjusted risks of mortality at 12 months (OR, 0.71; CI, 0.61–0.81), 3 months (OR, 0.72; CI, 0.61–0.85), and 1 month (OR, 0.61; CI, 0.48–0.79) and in the hospital (OR, 0.71; CI, 0.58–0.88) than level-1 SBP. The cumulative times at the other 3 SBP ranges (<100 mm Hg, 100–120 mm Hg, and ≥140 mm Hg) were not independent risk predictors of prognosis. Furthermore, Group M had lower 12-month mortality than Group L, which remained in the propensity-score matched and subgroup analyses.

Conclusions:

SBP at 120 mm Hg to 140 mm Hg was associated with decreased adverse outcomes. Randomized trials are required to determine whether the outcomes in critically ill patients improve with early maintenance of a SBP level at 120 mm Hg to 140 mm Hg.
Irisin Protects Against LPS-Stressed Cardiac Damage Through Inhibiting Inflammation, Apoptosis, and Pyroptosis
Authors: Li Qian , Zhang Mingming , Zhao Ying , Dong Maolong
Published: March 2021

Septic cardiac dysfunction remains a clinical problem due to its high morbidity and mortality. Uncontrolled cell death and excessive inflammatory response are closely related to sepsis-induced cardiac dysfunction. Irisin has been found to play cardioprotective roles in sepsis. However, there is enough uncertainty in the mechanism of irisin-mediated cardioprotection. We hypothesized that irisin may ameliorate myocardial dysfunction via reducing cardiac apoptosis, pyroptosis, and inflammation during LPS-induced sepsis. Mice were subjected to LPS with or without irisin treatment. After stimuli of LPS, the function of myocardium was distinctly impaired, which was closely related to increased level of apoptosis (decreased expression of Bcl-2 and elevated expression of Caspase-3 and Bax), pyroptosis (increased expression of Caspase1, NLR family pyrin domain containing 3 (NLRP3), and gasdermin D) and inflammatory mediators (increased level of IL-1β, TNF-α, and IL-6). This process is consistent with increased toll-like receptor 4 (TLR4)/nuclear factor-kappa B signal, apoptotic signal, and NLRP3-mediated pyroptotic signal. Activation of apoptosis and pyroptosis enhanced the expression of proinflammatory cytokines and further exacerbated septic myocardial damage. However, irisin can inhibit the expression of TLR4 and its downstream signaling molecules and also lower the level of apoptosis and pyroptosis. Besides, similar results were also found in vitro model of LPS-induced H9c2 cardiomyocyte injury. In general, irisin suppressed inflammation, apoptosis, and pyroptosis by blocking the TLR4 and NLRP3 inflammasome signalings to mitigate myocardial dysfunction in sepsis.
Estrogen Enhances the Microvascular Reactivity Through RhoA-ROCK Pathway in Female Mice During Hemorrhagic Shock
Authors: Yue Yun-Xue , Zhai Jia-Yi , Du Hui-Bo , Jiang Li-Na , Zhang Li-Min , Wang Chen , Zhao Zhen-Ao , Zhang Chun-Hui , Zhao Zi-Gang
Published: March 2021

Vascular hypo-reactivity plays a critical role inducing organ injury during hemorrhagic shock. 17β-estradiol (E2) can induce vasodilation to increase blood flow in various vascular beds. This study observed whether E2 can restore vascular hypo-reactivity induced by hemorrhagic shock, and whether E2 effects are associated with RhoA-Rho kinase (ROCK)-myosin light chain kinase phosphatase (MLCP) pathway. The hemorrhagic shock model (40 ± 2 mm Hg for 1 h, resuscitation for 4 h) was established in ovary intact sham operation (OVI), ovariectomized (OVX), and OVX plus E2 supplement female mice. Intestinal microvascular loop was used to assess blood flow in vivo, mRNA expression and vascular reactivity in vitro. Hemorrhagic shock significantly reduced norepinephrine microvascular reactivity. Decreased microvascular reactivity was exacerbated by OVX and reversed by E2 supplement. U-46619 (RhoA agonist) increased microvascular reactivity, and C3 transferase (an ADP ribosyl transferase that selectively induces RhoA ribosylation) or Y-27632 (ROCK inhibitor) inhibited sham mice microvascular reactivity. Similarly, U-46619 increased microvascular reactivity in OVI and OVX mice following hemorrhagic shock, which was abolished by Y-27632 or concomitant incubation of okadaic acid (OA) (MLCP inhibitor) and Y-27632. In OVX plus E2 supplement mice with hemorrhagic shock, Y-27632 inhibited microvascular reactivity, which was abolished by concomitant U-46619 application. Lastly, hemorrhagic shock remarkably decreased intestinal loop blood flow, RhoA and ROCK mRNA expressions in vascular tissues in OVX females, but not in OVI females, which were reversed by E2 supplement. These results indicate that estrogen improves microvascular reactivity during hemorrhagic shock, and RhoA-ROCK signaling pathway may mediate E2 effects.
Could Soluble Endothelial Protein C Receptor Levels Recognize SARS-CoV2-Positive Patients Requiring Hospitalization?
Authors: Vassiliou Alice G. , Keskinidou Chrysi , Jahaj Edison , Gallos Parisis , Zacharis Alexandros , Athanasiou Nikolaos , Tsipilis Stamatios , Mastora Zafeiria , Dimopoulou Ioanna , Kotanidou Anastasia , Orfanos Stylianos E.
Published: March 2021

Introduction:

The endothelial protein C receptor (EPCR) is a protein that regulates the protein C anticoagulant and anti-inflammatory pathways. A soluble form of EPCR (sEPCR) circulates in plasma and inhibits activated protein C (APC) activities. The clinical impact of sEPCR and its involvement in COVID-19 has not been explored. In this study, we investigated whether sEPCR levels were related to COVID-19 patients’ requirement for hospitalization.

Methods:

Plasma sEPCR levels were measured on hospital admission in 84 COVID-19 patients, and in 11 non-hospitalized SARS-CoV2-positive patients approximately 6 days after reported manifestation of their symptoms. Multiple logistic regression analysis was performed to identify potential risk factors for hospitalization and receiver operating characteristic (ROC) curves were generated to assess their value.

Results:

In our cohort, hospitalized patients had considerably higher sEPCR levels upon admission compared with outpatients [107.5 (76.7–156.3) vs. 44.6 (12.1–84.4) ng/mL; P < 0.0001)]. The ROC curve using hospitalization as the classification variable and sEPCR levels as the prognostic variable generated an area under the curve at 0.845 (95% CI = 0.710–0.981, P < 0.001). Additionally, we investigated the predictive value of sEPCR combined with BMI, age, or D-dimers.

Conclusions:

In our cohort, sEPCR levels in COVID-19 patients upon hospital admission appear considerably elevated compared with outpatients; this could lead to impaired APC activities and might contribute to the pro-coagulant phenotype reported in such patients. sEPCR measurement might be useful as a point-of-care test in SARS-CoV2-positive patients.
Detection of Mutations Associated with Variants of Concern Via High Throughput Sequencing of SARS-CoV-2 Isolated from NYC Wastewater
Authors: Smyth Davida S. , Trujillo Monica , Cheung Kristen , Gao Anna , Hoxie Irene , Kannoly Sherin , Kubota Nanami , Markman Michelle , San Kaung Myat , Sompanya Geena , Dennehy John J.
Published: March 2021

Monitoring SARS-CoV-2 genetic diversity is strongly indicated because diversifying selection may lead to the emergence of novel variants resistant to naturally acquired or vaccine-induced immunity. To date, most data on SARS-CoV-2 genetic diversity has come from the sequencing of clinical samples, but such studies may suffer limitations due to costs and throughput. Wastewater-based epidemiology may provide an alternative and complementary approach for monitoring communities for novel variants. Given that SARS-CoV-2 can infect the cells of the human gut and is found in high concentrations in feces, wastewater may be a valuable source of SARS-CoV-2 RNA, which can be deep sequenced to provide information on the circulating variants in a community. Here we describe a safe, affordable protocol for the sequencing of SARS-CoV-2 RNA using high-throughput Illumina sequencing technology. Our targeted sequencing approach revealed the presence of mutations associated with several Variants of Concern at appreciable frequencies. Our work demonstrates that wastewater-based SARS-CoV-2 sequencing can inform surveillance efforts monitoring the community spread of SARS-CoV-2 Variants of Concern and detect the appearance of novel emerging variants more cheaply, safely, and efficiently than the sequencing of individual clinical samples.

IMPORTANCE

The SARS-CoV-2 pandemic has caused millions of deaths around the world as countries struggle to contain infections. The pandemic will not end until herd immunity is reached, that is, when most of the population has either recovered from SARS-CoV-2 infection or is vaccinated against SARS-CoV-2. However, the emergence of new SARS-CoV-2 variants of concern threatens to erase gains. Emerging new variants may re-infect persons who have recovered from COVID-19 or may evade vaccine-induced immunity. However, scaling up SARS-CoV-2 genetic sequencing to monitor Variants of Concern in communities around the world is challenging. Wastewater-based sequencing of SARS-CoV-2 RNA can be used to monitor the presence of emerging variants in large communities to enact control measures to minimize the spread of these variants. We describe here the identification of alleles associated with several variants of concern in wastewater obtained from NYC watersheds.
A baton-relay and proofreading mechanism for selective ER retrieval signal capture by the KDEL receptor
Authors: Gerondopoulos Andreas , Bräuer Philipp , Sobajima Tomoaki , Wu Zhiyi , Parker Joanne L. , Biggin Philip C. , Barr Francis A. , Newstead Simon
Published: March 2021

The KDEL-retrieval pathway captures escaped ER proteins with a KDEL or variant C-terminal signal at acidic pH in the Golgi and releases them at neutral pH in the ER. To address the mechanism of signal binding and the molecular basis for differences in signal affinity, we determined the HDEL and RDEL bound structures of the KDEL-receptor. Affinity differences are explained by interactions between the variable −4 position of the signal and W120, whereas initial capture of retrieval signals by their carboxyl-terminus is mediated by a baton-relay mechanism involving a series of conserved arginine residues in the receptor. This explains how the signal is first captured and then pulled into the binding cavity. During capture, retrieval signals undergo a selective proofreading step involving two gatekeeper residues D50 and E117 in the receptor. These mechanisms operate upstream of the pH-dependent closure of the receptor and explain the selectivity of the KDEL-retrieval pathway.
A novel approach to comparative RNA-Seq does not support a conserved set of genes underlying animal regeneration
Authors: Sierra Noemie , Olsman Noah , Yi Lynn , Pachter Lior , Goentoro Lea , Gold David A.
Published: March 2021

Molecular studies of animal regeneration typically focus on conserved genes and signaling pathways that underlie morphogenesis. To date, a holistic analysis of gene expression across animals has not been attempted, as it presents a suite of problems related to differences in experimental design and gene homology. By combining orthology analysis with a novel statistical method for testing gene enrichment across large datasets, we are able to test whether biological processes across organisms share transcriptional regulation. We applied this method to six publicly available RNA-seq datasets from diverse examples of animal regeneration. We recovered 160 conserved orthologous gene clusters, which are enriched in structural genes as opposed to those regulating morphogenesis. A breakdown of gene presence/absence provides only limited support for the conservation of pathways typically implicated in regeneration, such as Wnt signaling and cell pluripotency. Specifically, these pathways are only conserved if we allow gene paralogs to be interchangeable through evolution. Overall, our analysis does not support the hypothesis that a shared set of ancestral genes underlie regeneration mechanisms in animals. The methods described in this paper will be broadly applicable for studying the genetic underpinnings of traits across distantly related organisms.
Over-expression of the Regulator of the Signaling of G Protein-Coupled Receptors RGS9-2 reduces the signaling elicited by the human histamine H3receptor in HEK-293T cells
Authors: Nieto-Alamilla Gustavo , Escamilla-Sánchez Juan , Arias-Montaño José-Antonio
Published: March 2021

In HEK-293T cells transiently transfected with the human histamine H3receptor (hH3R), we studied the effect of over-expressing the human RGS9-2 protein on H3R-mediated stimulation of [35S]-GTPγS binding and inhibition of forskolin-induced cAMP formation. Maximal specific binding (Bmax) of [3H]-N-methyl-histamine to cell membranes was 468 ± 12 and 442 ± 38 fmol/mg protein for HEK-293T-hH3R and HEK-293T-hH3R/hRGS9-2 cells, respectively, with dissociation constants (Kd) 2.57 nM and 3.38 nM. The H3R agonist immepip stimulated [35S]-GTPγS binding with similar potency and efficacy (Emax146.3 ± 4.4 % and 150.0 ± 5.3 % of basal, pEC508.57 ± 0.26 and 9.00 ± 0.33, respectively), but was significantly less efficacious to inhibit forskolin-induced cAMP accumulation in HEK-293T-hH3R/hRGS9 cells (−19.2 ± 5.3 versus −37.7 ± 5.1 % in HEK-293T-hH3R cells) with no significant difference in potency (pEC509.60 ± 0.14 and 9.07 ± 0.29, respectively). These results indicate that in HEK-293T cells hRGS9-2 regulates hH3R445signaling downstream G protein activation.
TWIST1 expression is associated with high-risk Neuroblastoma and promotes Primary and Metastatic Tumor Growth
Authors: Sepporta Maria-Vittoria , Praz Viviane , Bourloud Katia Balmas , Joseph Jean-Marc , Jauquier Nicolas , Riggi Nicolo’ , Nardou-Auderset Katya , Petit Audrey , Scoazec Jean-Yves , Sartelet Hervé , Renella Raffaele , Mühlethaler-Mottet Annick
Published: March 2021

The embryonic transcription factors TWIST1/2 are frequently overexpressed in cancer, acting as multifunctional oncogenes. Here we investigate their role in neuroblastoma (NB), a heterogeneous childhood malignancy ranging from spontaneous regression to dismal outcomes despite multimodal therapy. We first reveal the association of TWIST1 expression with poor survival and metastasis in primary NB, while TWIST2 correlates with good prognosis. Secondly, suppression of TWIST1 by CRISPR/Cas9 results in a reduction of tumor growth and metastasis colonization in immunocompromised mice. Moreover, TWIST1 knockout tumors display a less aggressive cellular morphology and a reduced disruption of the extracellular matrix (ECM) reticulin network. Additionally, we identify a TWIST1-mediated transcriptional program associated with dismal outcome in NB and involved in the control of pathways mainly linked to the signaling, migration, adhesion, the organization of the ECM, and the tumor cells versus tumor stroma crosstalk. Taken together, our findings suggest TWIST1 as novel therapeutic target in NB.
The plant circadian clock gene LHY influences Medicago truncatula nodulation
Authors: Achom M , Roy P , Lagunas B , Bonyadi-Pour R , Pardal AJ , Baxter L , Richmond B , Aschauer N , Fletcher E , Picot E , Rowson M , Blackwell J , Rich-Griffin C , Mysore KS , Wen J , Ott S , Carré IA , Gifford ML
Published: March 2021

Legumes house nitrogen-fixing endosymbiotic rhizobia in specialized polyploid cells within root nodules, which are factories of metabolic activity. We discovered that the circadian clock-associated transcriptional factor LATE ELONGATED HYPOCOTYL (LHY) affects nodulation in Medicago truncatula. By carrying out expression analysis of transcripts over time in nodules we found that the clock enables coordinated control of metabolic and regulatory processes linked to nitrogen fixation. Rhythmic transcripts in root nodules include a subset of Nodule-specific Cysteine Rich peptides (NCRs) that have the LHY-bound conserved Evening Element in their promoters. Until now, studies have suggested that NCRs act to regulate bacteroid differentiation and keep the rhizobial population in check. However, these conclusions came from the study of a few members of this very large gene family that has complex diversified spatio-temporal expression. We suggest that rhythmic expression of NCRs may be important for temporal coordination of bacterial activity with the rhythms of the plant host, in order to ensure optimal symbiosis.

Highlights

The circadian clock-associated transcriptional factor LATE ELONGATED HYPOCOTYL (LHY) impacts on successful Medicago truncatula-rhizobial symbiosis

The plant clock coordinates rhythmic patterns of metabolic and regulatory activity in nodules and drives rhythmic expression of a subset of Nodule-specific Cysteine Rich (NCR) genes.

Rhythmic expression of NCRs may be important for temporal coordination of bacterial activity with plant host rhythms to ensure optimal symbiosis.
NHE6-Depletion Corrects ApoE4-Mediated Synaptic Impairments and Reduces Amyloid Plaque Load
Authors: Pohlkamp Theresa , Xian Xunde , Wong Connie H , Durakoglugil Murat , Werthmann Gordon C , Saido Takaomi , Evers Bret M , White Charles L , Connor Jade , Hammer Robert E , Herz Joachim
Published: March 2021

Apolipoprotein E4 (ApoE4) is the most important and prevalent risk factor for late-onset Alzheimer’s disease (AD). The isoelectric point of ApoE4 matches the pH of the early endosome (EE), causing its delayed dissociation from ApoE receptors and hence impaired endolysosomal trafficking, disruption of synaptic homeostasis and reduced amyloid clearance. We have shown that enhancing endosomal acidification by inhibiting the EE-specific sodium-hydrogen exchanger NHE6 restores vesicular trafficking and normalizes synaptic homeostasis. Remarkably and unexpectedly, loss of NHE6 effectively suppressed amyloid deposition even in the absence of ApoE4, suggesting that accelerated acidification of early endosomes caused by the absence of NHE6 occludes the effect of ApoE on amyloid plaque formation. NHE6 suppression or inhibition may thus be a universal, ApoE-independent approach to prevent amyloid buildup in the brain. These findings suggest a novel therapeutic approach for the prevention of AD by which partial NHE6 inhibition reverses the ApoE4 induced endolysosomal trafficking defect and reduces amyloid.
Metabolic-sensing in AgRP neurons integrates homeostatic state with dopamine signalling in the striatum
Authors: Reichenbach Alex , Clarke Rachel E , Stark Romana , Lockie Sarah Haas , Mequinion Mathieu , Reed Felicia , Rawlinson Sasha , Dempsey Harry , Sepehrizadeh Tara , DeVeer Michael , Munder Astrid C , Nunez-Iglesias Juan , Spanswick David C. , Mynatt Randall , Kravitz Alexxai V. , Dayas Christopher V. , Brown Robyn , Andrews Zane B.
Published: March 2021

Hunger increases the motivation of an organism to seek out and consume highly palatable energy dense foods. While hunger-sensing Agouti-related peptide (AgRP) neurons influence this process, whether metabolic detection of homeostatic state via metabolic sensing in AgRP neurons potentiates motivation through the midbrain dopamine system is unexplored. Here, we used the AgRP-specific deletion of carnitine acetyltransferase (Crat), a metabolic enzyme regulating glucose and fatty acid oxidation, as a model of impaired metabolic-sensing in AgRP neurons. We then tested the hypothesis that appropriate metabolic-sensing in AgRP neurons is required to increase food reward motivation by modulating accumbal or striatal dopamine release. Electrophysiological studies confirm that Crat deletion in AgRP neurons (KO) impairs normalex vivoglucose-sensing, andin vivophotometry experiments show that AgRP neurons in KO mice do not exhibit normal responses to repeated palatable food presentation and consumption, highlighting that this model is appropriate to test the hypothesis. Fiber photometry experiments, using the dopamine sensor GRAB-DA, revealed that impaired metabolic-sensing reduces acute dopamine release (seconds) in the nucleus accumbens, but not the dorsal striatum, to palatable food consumption and during operant responding. Positron electron tomography (PET) methods indicated that impaired metabolic-sensing in AgRP neurons suppressed radiolabelled 18F-fDOPA accumulation after ∼30 minutes in the dorsal striatum but not the ventral striatum, suggesting a role for AgRP neurons to restrict a long term post-ingestive dopamine response in the dorsal striatum. Finally, impaired metabolic-sensing in AgRP neurons suppresses motivated operant responding for sucrose rewards. Notably, these behavioural effects are potentiated in the hungry state and therefore highlight that metabolic-sensing in AgRP neurons is required for the appropriate temporal integration and transmission of homeostatic hunger-sensing to dopamine signalling in the striatum.
Grp78 alleviates sodium iodate-induced retinal cell injury in vivo and in vitro
Authors: Shuang Jiang , Yongpeng Guo , Ning Yi , Hongdan Li , Hua Liu
Published: March 2021

Objective

Glucose-regulated protein 78 (Grp78) has been regarded as a main member of the endoplasmic reticulum proteins, Grp78 could protect cells from apoptosis under stress conditions. However, whether Grp78 could protect retinal pigment epithelium (RPE) cells from oxidative injury and then protect retinas from morphological changes and functional abnormalities remain undetermined. Here, we try to explore the effect of Grp78 on retinal cell injury induced by sodium iodate in vivo and in vitro.

Methods

To investigate whether Grp78 has a protective effect on RPE injury in vitro, human retinal pigment epithelium (ARPE-19) cells were treated with sodium iodate. The cell proliferation, morphology, apoptosis and ROS production assays were detected. In vivo, We established sodium iodate-induced retinal injury model in mice by intravenous injection of sodium iodate into tail vein. After that, we examined the morphology and function of retina in mice by fundus photography, OCT and ERG. Finally, we removed the retina of mice for histological examination.

Results

Grp78 significantly inhibited sodium iodate-induced reactive oxygen species (ROS), and decreased apoptosis of RPE in vitro. Furthermore, Grp78 significantly decreased the apoptosis of retinal cells in vivo, resulting in the inhibition of morphological changes of retina, and improving the function of retina. The underlying mechanisms included inhibited caspase3 and Nos, and increased expression of Bcl2, thereby protecting RPE from SI-induced ROS and apoptosis.

Conclusion

Grp78 could reduce the injury of retinal cells induced by sodium iodate in vitro and in vivo. These findings suggested Grp78 may become a new therapeutic target for retinal injury in clinical practice.
Recurrence quantification analysis of heart rate variability is a COVID-safe alternative to gas analysis in the detection of metabolic thresholds
Authors: Zimatore G. , Falcioni L. , Gallotta M.C. , Bonavolontà V. , Campanella M. , De Spirito M. , Guidetti L. , Baldari C.
Published: March 2021

The first aim of the study was to verify if in individuals with different physical fitness levels the Recurrence Quantification Analysis (RQA) of Heart Rate Variability (HRV) time series could be an alternative to Gas Exchange (GE) analysis in the determination of metabolic thresholds. The second aim was to investigate the validity of the RQA method compared to the GE method in thresholds detection. The two metabolic thresholds were estimated in thirty young individuals during Cardiopulmonary Exercise Testing on a cycle-ergometer and HR, VO2and Workload were measured by the two different methods (RQA and GE methods). RM ANOVA was used to assess main effects of methods and methods-by-groups interaction effects for HR, VO2and Workload at the aerobic (AerT) and the anaerobic (AnT) thresholds. Validity of the RQA at both thresholds was assessed for HR, VO2and Workload by Ordinary Least Products (OLP) regression analysis, Typical Percentage Errors (TE), Intraclass Correlation Coefficients (ICC) and the Bland Altman plots. No methods-by-groups interaction effects were detected for HR, VO2and Workload at the AerT and the AnT. The OLP regression analysis showed that at both thresholds RQA and GE methods had very strong correlations (r>0.8) in all variables (HR, VO2and Workload). Slope and intercept values always included the 1 and the 0, respectively. At the AerT the TE ranged from 4.02% to 10.47% (HR and Workload, respectively) and in all variables ICC values were excellent (≥0.85). At the AnT the TE ranged from 2.61% to 6.64% (HR and Workload, respectively) and in all variables ICC values were excellent (≥0.89). Our results suggest that the RQA of HRV time series is a COVID-safe approach for the determination of metabolic thresholds in individuals with different physical fitness levels, therefore, it can be used as a valid method for threshold detection alternative to gas analysis.
SNP- Based Assessment of Genetic Purity and Diversity in Maize Hybrid Breeding
Authors: Josia Chimwemwe , Mashingaidze Kingstone , Amelework Assefa B. , Kondwakwenda Aleck , Musvosvi Cousin , Sibiya Julia
Published: March 2021

Assessment of genetic purity of inbred lines and their resultant F1hybrids is an essential quality control check in maize hybrid breeding, variety release and seed production. In this study, genetic purity, parent-offspring relationship and diversity among the inbred lines were assessed using 92 single-nucleotide polymorphism (SNP) markers. A total of 188 maize genotypes, comprising of 26 inbred lines, four doubled haploid (DH) lines and 158 single-cross maize hybrids were investigated in this study using Kompetitive Allele Specific Polymerase Chain Reaction (KASP) genotyping assays. The bi-allelic data was analyzed for genetic purity and diversity parameters using GenAlex software. The SNP markers were highly polymorphic and 90% had polymorphic information content (PIC) values of > 0.3. Pairwise genetic distances among the lines ranged from 0.05 to 0.56, indicating a high level of dissimilarity among the inbred lines. Maximum genetic distance of (0.56) was observed for CKDHL0089, CML443 and CB323, while the lowest (0.05) was between I-42 and I-40. The majority (67%) of the inbred lines studied were genetically pure with residual heterozygosity of <5%, while only 33% were had heterozygosity levels of >5%. Inbred lines, which were not pure, require purification through further inbreeding. Cluster analysis partitioned the lines into three distinct genetic clusters with the potential to contribute new beneficial alleles to the maize breeding program. Out of the 68 hybrids (43%) that passed the parent-offspring test, seven hybrids namely; SCHP29, SCHP95, SCHP94, SCHP134, SCHP44, SCHP114 and SCHP126, were selected as potential candidates for further evaluation and release due to their outstanding yield performance.
The involvement of type IV pili and phytochrome in gliding motility, lateral motility and phototaxis of the cyanobacteriumPhormidium lacuna
Authors: Lamparter Tilman , Babian Jennifer , Fröhlich Katrin , Mielke Marion , Weber Nora , Wunsch Nadja , Zais Finn , Schulz Kevin , Aschmann Vera , Spohrer Nina , Krauß Norbert
Published: March 2021

Phormidium lacuna, a filamentous cyanobacterium without heterocysts, can be transformed by natural transformation. These filaments are motile on agar and other surfaces and display rapid lateral movements in liquid culture. Furthermore, they exhibit phototactic response under vertical illumination in Petri dishes. We generated mutants in which a KanR resistance cassette was integrated in the phytochrome genecphAand in various genes of the type IV pilin apparatus.pilM, pilN, pilQandpilTmutants were defective in all three responses, indicating that type IV pili are involved in all three kinds of motility. Rapid movements of wild type in liquid culture requires an extracellular matrix with type IV pili as central player.pilBmutants are only partially blocked in their responses.pilBis the proposed ATPase for expelling of the filament. In the mutant, this function could be overtaken by an alternative protein, likepilT, which regularly mediates retraction of pili. ThecphAmutant revealed a significantly reduced phototactic response towards red light. We assume that together with other photoreceptors, CphA regulates the phototactic response by down regulation of surface attachment.
Salty surfaces deter feeding in a blood-sucking disease vector
Authors: Pontes G , Latorre-Estivalis JM , Gutiérrez ML , Cano A , Berón de Astrada M , Lorenzo MG , Barrozo RB
Published: March 2021

Salts are essential nutrients required for many physiological processes, and deficient or excessive salt results in adverse health problems. Taste is the ultimate sensory modality involved in resource quality assessment, resulting in acceptance or rejection. Here, we show that detection of high-salt substrates by a salt-sensitive antennal gustatory receptor neuron, S1-GRN, results in feeding avoidance in the hematophagous bugRhodnius prolixus. Knock-down of two antennal-expressed amiloride-sensitive pickpocket channel receptors (PPKs;RproPPK014276andRproPPK28) using RNA interference, prevents avoidance of bugs to high-salt substrates. Tracing antennal GRNs to the central nervous system reveals the antennal lobes as a gustatory processing center. The identification of the gustatory basis of high-salt detection in a blood feeder provides novel targets to prevent biting and feeding, as well as to promote substrate avoidance in a relevant disease vector.

Significance Statement

Detection of aversive gustatory stimuli induces avoidance responses in animals. Avoidance acquires particular interest if it reduces the biting rates of blood-feeding insects of medical relevance. Here we describe the molecular and physiological basis of high-salt detection in the blood-sucking disease vectorRhodnius prolixus. We show that detection of high-salt substrates through two PPK receptors expressed in an antennal gustatory receptor neuron produces feeding avoidance. Understanding these gustatory-driven aversive responses allows the hitherto overlooked use of gustatory molecules as a complement to known olfactory repellents.