bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification of TBX15 as an adipose master trans regulator of abdominal obesity genes
Authors: Pan David Z. , Miao Zong , Comenho Caroline , Rajkumar Sandhya , Koka Amogha , Lee Seung Hyuk T. , Alvarez Marcus , Kaminska Dorota , Ko Arthur , Sinsheimer Janet S. , Mohlke Karen L. , Mancuso Nicholas , Muñoz-Hernandez Linda Liliana , Herrera-Hernandez Miguel , Tusié-Luna Maria Teresa , Aguilar-Salinas Carlos , Pietiläinen Kirsi H. , Pihlajamäki Jussi , Laakso Markku , Garske Kristina M. , Pajukanta Päivi
Published: August 2021

Background

Obesity predisposes individuals to multiple cardiometabolic disorders, including type 2 diabetes (T2D). As body mass index (BMI) cannot reliably differentiate fat from lean mass, the metabolically detrimental abdominal obesity has been estimated using waist-hip ratio (WHR). Waist-hip ratio adjusted for body mass index (WHRadjBMI) in turn is a well-established sex-specific marker for abdominal fat and adiposity, and a predictor of adverse metabolic outcomes, such as T2D. However, the underlying genes and regulatory mechanisms orchestrating the sex differences in obesity and body fat distribution in humans are not well understood.

Methods

We searched for genetic master regulators of WHRadjBMI by employing integrative genomics approaches on human subcutaneous adipose RNA sequencing (RNA-seq) data (n ~ 1400) and WHRadjBMI GWAS data (n ~ 700,000) from the WHRadjBMI GWAS cohorts and the UK Biobank (UKB), using co-expression network, transcriptome-wide association study (TWAS), and polygenic risk score (PRS) approaches. Finally, we functionally verified our genomic results using gene knockdown experiments in a human primary cell type that is critical for adipose tissue function.

Results

Here, we identified an adipose gene co-expression network that contains 35 obesity GWAS genes and explains a significant amount of polygenic risk for abdominal obesity and T2D in the UKB (n = 392,551) in a sex-dependent way. We showed that this network is preserved in the adipose tissue data from the Finnish Kuopio Obesity Study and Mexican Obesity Study. The network is controlled by a novel adipose master transcription factor (TF), TBX15, a WHRadjBMI GWAS gene that regulates the network in trans. Knockdown of TBX15 in human primary preadipocytes resulted in changes in expression of 130 network genes, including the key adipose TFs, PPARG and KLF15, which were significantly impacted (FDR < 0.05), thus functionally verifying the trans regulatory effect of TBX15 on the WHRadjBMI co-expression network.

Conclusions

Our study discovers a novel key function for the TBX15 TF in trans regulating an adipose co-expression network of 347 adipose, mitochondrial, and metabolically important genes, including PPARG, KLF15, PPARA, ADIPOQ, and 35 obesity GWAS genes. Thus, based on our converging genomic, transcriptional, and functional evidence, we interpret the role of TBX15 to be a main transcriptional regulator in the adipose tissue and discover its importance in human abdominal obesity.
Teleneurorehabilitation program (virtual reality) for patients with balance disorders: descriptive study
Authors: Maldonado-Díaz Marcos , Vargas Patricia , Vasquez Ricardo , Gonzalez-Seguel Felipe , Rivero Betel , Hidalgo-Cabalín Viviane , Gutierrez-Panchana Tania
Published: August 2021

Background

Balance disorders are common in patients with neurological or vestibular diseases. Telerehabilitation program is a treatment to be as safe as conventional treatment. One of the most used methods to perform telerehabilitation is the incorporation of Virtual Reality. In general, rehabilitation programs train predictive postural control, so the patient does not always acquire the necessary autonomy to react to situations of instability. On the other hand, the objective and systematic supervision and measurement of these programs is limited, making it necessary to create clinical protocols with precise and measurable rehabilitation objectives. This study present the training selection methodology and clinical protocol for patients with balance disorders inserted in a Telerehabilitation Program based on Virtual Reality.

Methods

Descriptive study where physiotherapists were trained to use RehaMetrics®. To evaluate their level of agreement in the selection of the exercise clusters developed, the Interobserver Reliability was measured through the kappa statistic. Subsequently, the exercises were applied to a group of patients recruited with sedentary trunk control (Berg Balance Scale = 3 points in item 3), mild or normal cognitive level (Montreal Cognitive Assessment> 21 points), and prescribed for tele-rehabilitation by a doctor.

Results

The agreement among the expert physiotherapists irrespective of the cluster exceeds 80%, which indicates a very good strength of agreement, while the novices reached a level of agreement of 45%, which suggests a moderate strength of agreement. All clinical outcomes showed statistically significant differences between the median times, as did the Maximum Width Left Side (MWLS) (cm). The average number of minutes of training was 485.81 (SD 246.49 min), and the number of sessions performed during the 4 weeks of intervention was 17 (SD 7.15 sessions).

Conclusions

This analysis what had excellent interobserver reliability with trained physiotherapists. Regarding the second phase of the study, the results show a statistically significant difference between the initial and final evaluation of the clinical tests, which could result in better performance in aspects such as: balance, gait functionality, meter walked and cognition. Telerehabilitation Program based on Virtual Reality is an excellent alternative to provide continuity of treatment to patients with balance disorders.
Addressing vaccine hesitancy and access barriers to achieve persistent progress in Israel’s COVID-19 vaccination program
Authors: Rosen Bruce , Waitzberg Ruth , Israeli Avi , Hartal Michael , Davidovitch Nadav
Published: August 2021

As of March 31, 2021, Israel had administered 116 doses of vaccine for COVID-19 per 100 population (of any age) – far more than any other OECD country. It was also ahead of other OECD countries in terms of the share of the population that had received at least one vaccination (61%) and the share that had been fully vaccinated (55%). Among Israelis aged 16 and over, the comparable figures were 81 and 74%, respectively. In light of this, the objectives of this article are:

To describe and analyze the vaccination uptake through the end of March 2021

To identify behavioral and other barriers that likely affected desire or ability to be vaccinated

To describe the efforts undertaken to overcome those barriers

Israel’s vaccination campaign was launched on December 20, and within 2.5 weeks, 20% of Israelis had received their first dose. Afterwards, the pace slowed. It took an additional 4 weeks to increase from 20 to 40% and yet another 6 weeks to increase from 40 to 60%. Initially, uptake was low among young adults, and two religious/cultural minority groups - ultra-Orthodox Jews and Israeli Arabs, but their uptake increased markedly over time.

In the first quarter of 2021, Israel had to enhance access to the vaccine, address a moderate amount of vaccine hesitancy in its general population, and also address more intense pockets of vaccine hesitancy among young adults and religious/cultural minority groups. A continued high rate of infection during the months of February and March, despite broad vaccination coverage at the time, created confusion about vaccine effectiveness, which in turn contributed to vaccine hesitancy. Among Israeli Arabs, some residents of smaller villages encountered difficulties in reaching vaccination sites, and that also slowed the rate of vaccination.

The challenges were addressed via a mix of messaging, incentives, extensions to the initial vaccine delivery system, and other measures. Many of the measures addressed the general population, while others were targeted at subgroups with below-average vaccination rates. Once the early adopters had been vaccinated, it took hard, creative work to increase population coverage from 40 to 60% and beyond.

Significantly, some of the capacities and strategies that helped Israel address vaccine hesitancy and geographic access barriers are different from those that enabled it to procure, distribute and administer the vaccines. Some of these strategies are likely to be relevant to other countries as they progress from the challenges of securing an adequate vaccine supply and streamlining distribution to the challenge of encouraging vaccine uptake.
An extended approach to value chain analysis
Authors: Knez Klemen , Jaklič Andreja , Stare Metka
Published: August 2021

In the article, we propose a comprehensive methodology of value chain analysis in the international input–output framework that introduces a new measure of value chain participation and an extended typology of value chains, with the novel inclusion of domestic value chain to address the extent of fragmentation of purely domestic production. This allows for the simultaneous analysis of both global and domestic production fragmentation, the complex patterns of their evolution and their impact on economic development. The main contribution of the proposed methodology is conceptual: it permits the measurement of all value chain paths that pass through each country-sector from production to final consumption, whether the path includes downstream linkages, upstream linkages or their combination. Empirical application of this methodology shows the importance of including domestic fragmentation in value chain analysis: The fragmentation of both global and domestic levels of production has a significant positive correlation with economic growth. This implies that the effects of global production fragmentation must be analysed together with the changing structure of the fragmentation of domestic production to obtain the whole picture, one that might provide important information for policymaking and industrial policy.
Tendon grafts with preserved muscle demonstrate similar biomechanical properties to tendon grafts stripped of muscular attachments: a biomechanical evaluation in a porcine model
Authors: Funchal Luis Fernando Zukanovich , Astur Diego Costa , Pizzolatti André Luiz Almeida , Grimaldi Arthur Paiva , Jimenez Andrew Esteban , Moré Ari Digiácomo Ocampo , de Mello Roesler Carlos Rodrigo , Cohen Moises
Published: August 2021

Purpose

(1) To evaluate the biomechanical properties of a porcine flexor digitorum superficialis tendon graft with preserved muscle fibers and (2) to compare these results with the biomechanical properties of a porcine tendon graft after removal of associated muscle.

Methods

Eighty-two porcine forelegs were dissected and the flexor digitorum superficialis muscle tendons were harvested. The study comprised of two groups: Group 1 (G1), harvested tendon with preserved muscle tissue; and Group 2 (G2), harvested contralateral tendon with removal of all muscle tissue. Tests in both groups were conducted using an electro-mechanical material testing machine (Instron, model 23-5S, Instron Corp., Canton, MA, USA) with a 500 N force transducer. Yield load, stiffness, and maximum load were evaluated and compared between groups.

Results

The behavior of the autografts during the tests followed the same stretching, deformation, and failure patterns as those observed in human autografts subjected to axial strain. There were no significant differences in the comparison between groups for ultimate load to failure (p = 0.105), stiffness (p = 0.097), and energy (p = 0.761).

Conclusion

In this porcine model biomechanical study, using autograft tendon with preserved muscle showed no statistically significant differences for yield load, stiffness, or maximum load compared to autograft tendon without preserved muscle. The preservation of muscle on the autograft tendon did not compromise the mechanical properties of the autograft.

Level of evidence

Level III Controlled laboratory study
Effect of thinning intensity on understory herbaceous diversity and biomass in mixed coniferous and broad-leaved forests of Changbai Mountain
Authors: Wang Gerong , Sun Yue , Zhou Mo , Guan Naiqian , Wang Yuwen , Jiang Runhua , Liu Zhiyu , Wu Mengjia , Xia Fucai
Published: August 2021

Background

Herbs are an important part of the forest ecosystem, and their diversity and biomass can reflect the restoration of vegetation after forest thinning disturbances. Based on the near-mature secondary coniferous and broad-leaved mixed forest in Jilin Province Forestry Experimental Zone, this study analyzed seasonal changes of species diversity and biomass of the understory herb layer after different intensities of thinning.

Results

The results showed that although the composition of herbaceous species and the ranking of importance values were affected by thinning intensity, they were mainly determined by seasonal changes. Across the entire growing season, the species with the highest importance values in thinning treatments included Carex pilosa, Aegopodium alpestre, Meehania urticifolia, and Filipendula palmata, which dominated the herb layer of the coniferous and broad-leaved mixed forest. The number of species, Margalef index, Shannon-Wiener index and Simpson index all had their highest values in May, and gradually decreased with months. Pielou index was roughly inverted “N” throughout the growing season. Thinning did not increase the species diversity. Thinning can promote the total biomass, above- and below-ground biomass. The number of plants per unit area and coverage were related to the total biomass, above- and below-ground biomass. The average height had a significantly positive correlation with herb biomass in May but not in July. However, it exerted a significantly negative correlation with herb biomass in September. The biomass in the same month increased with increasing thinning intensity. Total herb biomass, above- and below-ground biomass showed positive correlations with Shannon-Winner index, Simpson index and Pielou evenness index in May.

Conclusions

Thinning mainly changed the light environment in the forest, which would improve the plant diversity and biomass of herb layer in a short time. And different thinning intensity had different effects on the diversity of understory herb layer. The findings provide theoretical basis and reference for reasonable thinning and tending in coniferous and broad-leaved mixed forests.
ICPFuzzer: proprietary communication protocol fuzzing by using machine learning and feedback strategies
Authors: Lin Pei-Yi , Tien Chia-Wei , Huang Ting-Chun , Tien Chin-Wei
Published: August 2021

The fuzzing test is able to discover various vulnerabilities and has more chances to hit the zero-day targets. And ICS(Industrial control system) is currently facing huge security threats and requires security standards, like ISO 62443, to ensure the quality of the device. However, some industrial proprietary communication protocols can be customized and have complicated structures, the fuzzing system cannot quickly generate test data that adapt to various protocols. It also struggles to define the mutation field without having prior knowledge of the protocols. Therefore, we propose a fuzzing system named ICPFuzzer that uses LSTM(Long short-term memory) to learn the features of a protocol and generates mutated test data automatically. We also use the responses of testing and adjust the weight strategies to further test the device under testing (DUT) to find more data that cause unusual connection status. We verified the effectiveness of the approach by comparing with the open-source and commercial fuzzers. Furthermore, in a real case, we experimented with the DLMS/COSEM for a smart meter and found that the test data can cause a unusual response. In summary, ICPFuzzer is a black-box fuzzing system that can automatically execute the testing process and reveal vulnerabilities that interrupt and crash industrial control communication. Not only improves the quality of ICS but also improves safety.
Evaluation of the frequency and patterns of thyroid dysfunction in patients with metabolic syndrome
Authors: El-Hay Gehad Abd El-Shaker Abd , Argoon Salah Abdelazim , Mousa Naglaa Mohamed M. A.
Published: August 2021

Background

Metabolic syndrome (MetS) and thyroid dysfunction have a degree of close association, and each of them affects the other. Due to the associated cardiovascular events, MetS has increased morbidity and mortality. The study tried to detect the frequency of thyroid function in patients with MetS. This is a case control study that recruited 100 patients with MetS and 100 healthy control subjects.

Results

Patients with MetS had significantly higher body mass index and waist circumference. Also, frequency of thyroid dysfunction was significantly higher in MetS group (32% vs. 9%; P<0.001). The most frequent form of thyroid dysfunction was subclinical hypothyroidism: 21% of the MetS group and 6% of the control group. Out of the studied patients with MetS, 13 (13%) patients had three criteria, 55 (55%) patients had four criteria, and 32 (32%) patients had five criteria for MetS.

Conclusion

Patients with MetS are vulnerable to develop thyroid dysfunction mainly subclinical hypothyroidism. So, it is recommended to perform regular screening for those patients as regard thyroid dysfunction.
Amivantamab in EGFR Exon 20 Insertion–Mutated Non–Small-Cell Lung Cancer Progressing on Platinum Chemotherapy: Initial Results From the CHRYSALIS Phase I Study
Authors: Park Keunchil , Haura Eric B. , Leighl Natasha B. , Mitchell Paul , Shu Catherine A. , Girard Nicolas , Viteri Santiago , Han Ji-Youn , Kim Sang-We , Lee Chee Khoon , Sabari Joshua K. , Spira Alexander I. , Yang Tsung-Ying , Kim Dong-Wan , Lee Ki Hyeong , Sanborn Rachel E. , Trigo José , Goto Koichi , Lee Jong-Seok , Yang James Chih-Hsin , Govindan Ramaswamy , Bauml Joshua M. , Garrido Pilar , Krebs Matthew G. , Reckamp Karen L. , Xie John , Curtin Joshua C. , Haddish-Berhane Nahor , Roshak Amy , Millington Dawn , Lorenzini Patricia , Thayu Meena , Knoblauch Roland E. , Cho Byoung Chul
Published: August 2021

PURPOSE

Non–small-cell lung cancer (NSCLC) with epidermal growth factor receptor ( EGFR) exon 20 insertion (Exon20ins) mutations exhibits inherent resistance to approved tyrosine kinase inhibitors. Amivantamab, an EGFR-MET bispecific antibody with immune cell–directing activity, binds to each receptor's extracellular domain, bypassing resistance at the tyrosine kinase inhibitor binding site.

METHODS

CHRYSALIS is a phase I, open-label, dose-escalation, and dose-expansion study, which included a population with EGFR Exon20ins NSCLC. The primary end points were dose-limiting toxicity and overall response rate. We report findings from the postplatinum EGFR Exon20ins NSCLC population treated at the recommended phase II dose of 1,050 mg amivantamab (1,400 mg, ≥ 80 kg) given once weekly for the first 4 weeks and then once every 2 weeks starting at week 5.

RESULTS

In the efficacy population (n = 81), the median age was 62 years (range, 42-84 years); 40 patients (49%) were Asian, and the median number of previous lines of therapy was two (range, 1-7). The overall response rate was 40% (95% CI, 29 to 51), including three complete responses, with a median duration of response of 11.1 months (95% CI, 6.9 to not reached). The median progression-free survival was 8.3 months (95% CI, 6.5 to 10.9). In the safety population (n = 114), the most common adverse events were rash in 98 patients (86%), infusion-related reactions in 75 (66%), and paronychia in 51 (45%). The most common grade 3-4 adverse events were hypokalemia in six patients (5%) and rash, pulmonary embolism, diarrhea, and neutropenia in four (4%) each. Treatment-related dose reductions and discontinuations were reported in 13% and 4% of patients, respectively.

CONCLUSION

Amivantamab, via its novel mechanism of action, yielded robust and durable responses with tolerable safety in patients with EGFR Exon20ins mutations after progression on platinum-based chemotherapy.
Cancer Care in the Wake of a Cyberattack: How to Prepare and What to Expect
Authors: Ades Steven , Herrera Diego Adrianzen , Lahey Tim , Thomas Alissa A. , Jasra Sakshi , Barry Maura , Sprague Julian , Dittus Kim , Plante Timothy B. , Kelly Jamie , Kaufman Peter A. , Khan Farrah , Hammond Cory J. , Gernander Kelly , Parsons Polly , Holmes Chris
Published: August 2021

PURPOSE:

Cyberattacks targeting health care organizations are becoming more frequent and affect all aspects of care delivery. Cancer care is particularly susceptible to major disruptions because of the potential of immediate and long-term consequences for patients who often rely on timely diagnostic testing and regular administration of systemic therapy in addition to other local treatment modalities to cure or control their diseases. On October 28, 2020, a cyberattack was launched on the University of Vermont Health Network with wide-ranging consequences for oncology, including loss of access to all network intranet servers, e-mail communications, and the electronic medical record (EMR).

METHODS:

This review details the immediate challenges faced by hematology and oncology during the cyberattack. The impact and response on inpatient, outpatient, and special patient populations are described. Steps that other academic- and community-based oncology practices can take to lessen the brunt of such an assault are suggested.

RESULTS:

The two areas of immediate impact after the cyberattack were communications and lack of EMR access. The oncology-specific impact included loss of the individualized EMR chemotherapy plan templates and electronic safeguards built into multistep treatment preparation and delivery. With loss of access to schedules, basic patient information, encrypted communications platforms and radiology, and laboratory and pharmacy services, clinical outpatient care delivery was reduced by 40%. The infusion visit volume dropped by 52% in the first week and new patients could not access necessary services for timely diagnostic evaluation, requiring the creation of command centers to oversee ethical and transparent triage and allocation of systemic therapies and address new patient referrals. This included appropriate transfer of patients to alternate sites to minimize delays. Inpatient care including transitions of care was particularly challenging and addressing patient populations whose survival might be affected by delays in care.

CONCLUSION:

Oncology health care leaders and providers should be aware of the potential impact of a cyberattack on cancer care delivery and preventively develop processes to mitigate the impact.
Rescuing Henry George
Authors: Mueller Thomas Michael
Published: August 2021

Harold Hotelling’s work, from natural resources economics to optimal taxation, and from spatial to welfare economics, was deeply influenced by his Georgist affiliation and by a Georgist game that we now call Monopoly, which at the time was known as the Landlord’s Game. We explore this influence, its history, and the role it played in Hotelling’s work and ideas. We show that political beliefs deeply shaped Hotelling’s approach to economics and that the rules of the Landlord’s Game helped him think about economic mechanisms.
Amkoullel, the Fula Boy
Authors: Bâ Amadou Hampâté
Published: August 2021

Born in 1900 in French West Africa, Malian writer Amadou Hampâté Bâ was one of the towering figures in the literature of twentieth-century Francophone Africa. In Amkoullel, the Fula Boy, Bâ tells in striking detail the story of his youth, which was set against the aftermath of war between the Fula and Toucouleur peoples and the installation of French colonialism. A master storyteller, Bâ recounts pivotal moments of his life, and the lives of his powerful and large family, from his first encounter with the white commandant through the torturous imprisonment of his stepfather and to his forced attendance at French school. He also charts a larger story of life prior to and at the height of French colonialism: interethnic conflicts, the clash between colonial schools and Islamic education, and the central role indigenous African intermediaries and interpreters played in the functioning of the colonial administration. Engrossing and novelistic, Amkoullel, the Fula Boy is an unparalleled rendering of an individual and society under transition as they face the upheavals of colonialism.
Speaking for the People
Authors: Rifkin Mark
Published: August 2021

In Speaking for the People Mark Rifkin examines nineteenth-century Native writings to reframe contemporary debates around Indigenous recognition, refusal, and resurgence. Rifkin shows how works by Native authors (William Apess, Elias Boudinot, Sarah Winnemucca, and Zitkala-Ša) illustrate the intellectual labor involved in representing modes of Indigenous political identity and placemaking. These writers highlight the complex processes involved in negotiating the character, contours, and scope of Indigenous sovereignties under ongoing colonial occupation. Rifkin argues that attending to these writers' engagements with non-native publics helps provide further analytical tools for addressing the complexities of Indigenous governance on the ground—both then and now. Thinking about Native peoplehood and politics as a matter of form opens possibilities for addressing the difficult work involved in navigating among varied possibilities for conceptualizing and enacting peoplehood in the context of continuing settler intervention. As Rifkin demonstrates, attending to writings by these Indigenous intellectuals provides ways of understanding Native governance as a matter of deliberation, discussion, and debate, emphasizing the open-ended unfinishedness of self-determination.
Birthing Black Mothers
Authors: Nash Jennifer C.
Published: August 2021

In Birthing Black Mothers Black feminist theorist Jennifer C. Nash examines how the figure of the “Black mother” has become a powerful political category. “Mothering while Black” has become synonymous with crisis as well as a site of cultural interest, empathy, fascination, and support. Cast as suffering and traumatized by their proximity to Black death—especially through medical racism and state-sanctioned police violence—Black mothers are often rendered as one-dimensional symbols of tragic heroism. In contrast, Nash examines Black mothers’ self-representations and public performances of motherhood—including Black doulas and breastfeeding advocates alongside celebrities such as Beyoncé, Serena Williams, and Michelle Obama—that are not rooted in loss. Through cultural critique and in-depth interviews, Nash acknowledges the complexities of Black motherhood outside its use as political currency. Throughout, Nash imagines a Black feminist project that refuses the lure of locating the precarity of Black life in women and instead invites readers to theorize, organize, and dream into being new modes of Black motherhood.
Whiteness Interrupted
Authors: Bell Marcus
Published: August 2021

In Whiteness Interrupted Marcus Bell presents a revealing portrait of white teachers in majority-black schools in which he examines the limitations of understandings of how white racial identity is formed. Through in-depth interviews with dozens of white teachers from a racially segregated, urban school district in Upstate New York, Bell outlines how whiteness is constructed based on localized interactions and takes a different form in predominantly black spaces. He finds that in response to racial stress in a difficult teaching environment, white teachers conceptualized whiteness as a stigmatized category predicated on white victimization. When discussing race outside majority-black spaces, Bell's subjects characterized American society as postracial, in which race seldom affects outcomes. Conversely, in discussing their experiences within predominantly black spaces, they rejected the idea of white privilege, often angrily, and instead focused on what they saw as the racial privilege of blackness. Throughout, Bell underscores the significance of white victimization narratives in black spaces and their repercussions as the United States becomes a majority-minority society.
Lrig1 and Lrig3 cooperate to control Ret receptor signaling, sensory axonal growth and epidermal innervation
Authors: De Vincenti Ana Paula , Alsina Fernando C. , Ferrero Restelli Facundo , Hedman Håkan , Ledda Fernanda , Paratcha Gustavo
Published: August 2021

Negative feedback loops represent a regulatory mechanism that guarantees that signaling thresholds are compatible with a physiological response. Previously, we established that Lrig1 acts through this mechanism to inhibit Ret activity. However, it is unclear whether other Lrig family members play similar roles. Here, we show that Lrig1 and Lrig3 are co-expressed in Ret-positive mouse dorsal root ganglion (DRG) neurons. Lrig3, like Lrig1, interacts with Ret and inhibits GDNF/Ret signaling. Treatment of DRG neurons with GDNF ligands induces a significant increase in the expression of Lrig1 and Lrig3. Our findings show that, whereas a single deletion of either Lrig1 or Lrig3 fails to promote Ret-mediated axonal growth, haploinsufficiency of Lrig1 in Lrig3 mutants significantly potentiates Ret signaling and axonal growth of DRG neurons in response to GDNF ligands. We observe that Lrig1 and Lrig3 act redundantly to ensure proper cutaneous innervation of nonpeptidergic axons and behavioral sensitivity to cold, which correlates with a significant increase in the expression of the cold-responsive channel TrpA1. Together, our findings provide insights into the in vivo functions through which Lrig genes control morphology, connectivity and function in sensory neurons.
A comprehensive series of Irx cluster mutants reveals diverse roles in facial cartilage development
Authors: Farmer D'Juan T. , Patel Punam , Choi Rachelle , Liu Chih-Yu , Crump J. Gage
Published: August 2021

Proper function of the vertebrate skeleton requires the development of distinct articulating embryonic cartilages. Irx transcription factors are arranged in co-regulated clusters that are expressed in the developing skeletons of the face and appendages. IrxB cluster genes are required for the separation of toes in mice and formation of the hyoid joint in zebrafish, yet whether Irx genes have broader roles in skeletal development remains unclear. Here, we perform a comprehensive loss-of-function analysis of all 11 Irx genes in zebrafish. We uncover conserved requirements for IrxB genes in formation of the fish and mouse scapula. In the face, we find a requirement for IrxAb genes and irx7 in formation of anterior neural crest precursors of the jaw, and for IrxBa genes in formation of endodermal pouches and gill cartilages. We also observe extensive joint loss and cartilage fusions in animals with combinatorial losses of Irx clusters, with in vivo imaging revealing that at least some of these fusions arise through inappropriate chondrogenesis. Our analysis reveals diverse roles for Irx genes in the formation and later segmentation of the facial skeleton.
osr1couples intermediate mesoderm cell fate with temporal dynamics of vessel progenitor cell differentiation
Authors: Perens Elliot A. , Diaz Jessyka T. , Quesnel Agathe , Askary Amjad , Crump J. Gage , Yelon Deborah
Published: August 2021

Transcriptional regulatory networks refine gene expression boundaries to define the dimensions of organ progenitor territories. Kidney progenitors originate within the intermediate mesoderm (IM), but the pathways that establish the boundary between the IM and neighboring vessel progenitors are poorly understood. Here, we delineate roles for the zinc-finger transcription factor Osr1 in kidney and vessel progenitor development. Zebrafish osr1 mutants display decreased IM formation and premature emergence of lateral vessel progenitors (LVPs). These phenotypes contrast with the increased IM and absent LVPs observed with loss of the bHLH transcription factor Hand2, and loss of hand2 partially suppresses osr1 mutant phenotypes. hand2 and osr1 are expressed together in the posterior mesoderm, but osr1 expression decreases dramatically prior to LVP emergence. Overexpressing osr1 during this timeframe inhibits LVP development while enhancing IM formation, and can rescue the osr1 mutant phenotype. Together, our data demonstrate that osr1 modulates the extent of IM formation and the temporal dynamics of LVP development, suggesting that a balance between levels of osr1 and hand2 expression is essential to demarcate the kidney and vessel progenitor territories.
Shedding light on developmental ERK signaling with genetically encoded biosensors
Authors: Nakamura Akinobu , Goto Yuhei , Kondo Yohei , Aoki Kazuhiro
Published: August 2021

The extracellular signal-regulated kinase (ERK) pathway governs cell proliferation, differentiation and migration, and therefore plays key roles in various developmental and regenerative processes. Recent advances in genetically encoded fluorescent biosensors have unveiled hitherto unrecognized ERK activation dynamics in space and time and their functional importance mainly in cultured cells. However, ERK dynamics during embryonic development have still only been visualized in limited numbers of model organisms, and we are far from a sufficient understanding of the roles played by developmental ERK dynamics. In this Review, we first provide an overview of the biosensors used for visualization of ERK activity in live cells. Second, we highlight the applications of the biosensors to developmental studies of model organisms and discuss the current understanding of how ERK dynamics are encoded and decoded for cell fate decision-making.
ELAC2/RNaseZ-linked cardiac hypertrophy in Drosophila melanogaster
Authors: Migunova Ekaterina , Theophilopoulos Joanna , Mercadante Marisa , Men Jing , Zhou Chao , Dubrovsky Edward B.
Published: August 2021

A severe form of infantile cardiomyopathy (CM) has been linked to mutations in ELAC2, a highly conserved human gene. It encodes Zinc phosphodiesterase ELAC protein 2 (ELAC2), which plays an essential role in the production of mature tRNAs. To establish a causal connection between ELAC2 variants and CM, here we used the Drosophila melanogaster model organism, which carries the ELAC2 homolog RNaseZ. Even though RNaseZ and ELAC2 have diverged in some of their biological functions, our study demonstrates the use of the fly model to study the mechanism of ELAC2-related pathology. We established transgenic lines harboring RNaseZ with CM-linked mutations in the background of endogenous RNaseZ knockout. Importantly, we found that the phenotype of these flies is consistent with the pathological features in human patients. Specifically, expression of CM-linked variants in flies caused heart hypertrophy and led to reduction in cardiac contractility associated with a rare form of CM. This study provides first experimental evidence for the pathogenicity of CM-causing mutations in the ELAC2 protein, and the foundation to improve our understanding and diagnosis of this rare infantile disease.

This article has an associated First Person interview with the first author of the paper.
Identification of candidate miRNA biomarkers for facioscapulohumeral muscular dystrophy using DUX4-based mouse models
Authors: Nunes Andreia M. , Ramirez Monique , Jones Takako I. , Jones Peter L.
Published: August 2021

Facioscapulohumeral muscular dystrophy (FSHD) is caused by misexpression of DUX4 in skeletal myocytes. As DUX4 is the key therapeutic target in FSHD, surrogate biomarkers of DUX4 expression in skeletal muscle are critically needed for clinical trials. Although no natural animal models of FSHD exist, transgenic mice with inducible DUX4 expression in skeletal muscles rapidly develop myopathic phenotypes consistent with FSHD. Here, we established a new, more-accurate FSHD-like mouse model based on chronic DUX4 expression in a small fraction of skeletal myonuclei that develops pathology mimicking key aspects of FSHD across its lifespan. Utilizing this new aged mouse model and DUX4-inducible mouse models, we characterized the DUX4-related microRNA signatures in skeletal muscles, which represent potential biomarkers for FSHD. We found increased expression of miR-31-5p and miR-206 in muscles expressing different levels of DUX4 and displaying varying degrees of pathology. Importantly, miR-206 expression is significantly increased in serum samples from FSHD patients compared with healthy controls. Our data support miR-31-5p and miR-206 as new potential regulators of muscle pathology and miR-206 as a potential circulating biomarker for FSHD.

This article has an associated First Person interview with the first author of the paper.
A three-dimensional spheroid-specific role for Wnt–β-catenin and Eph–ephrin signaling in nasopharyngeal carcinoma cells
Authors: Yi Canhui , Lai Sook Ling , Tsang Chi Man , Artemenko Margarita , Shuen Tang Maggie Kei , Pang Stella W. , Lo Kwok Wai , Tsao Sai Wah , Wong Alice Sze Tsai
Published: August 2021

One of the greatest unmet needs hindering the successful treatment of nasopharyngeal carcinomas (NPCs) is for representative physiological and cost-effective models. Although Epstein–Barr virus (EBV) infection is consistently present in NPCs, most studies have focused on EBV-negative NPCs. For the first time, we established and analyzed three-dimensional (3D) spheroid models of EBV-positive and EBV-negative NPC cells and compared these to classical two-dimensional (2D) cultures in various aspects of tumor phenotype and drug responses. Compared to 2D monolayers, the 3D spheroids showed significant increases in migration capacity, stemness characteristics, hypoxia and drug resistance. Co-culture with endothelial cells, which mimics essential interactions in the tumor microenvironment, effectively enhanced spheroid dissemination. Furthermore, RNA sequencing revealed significant changes at the transcriptional level in 3D spheroids compared to expression in 2D monolayers. In particular, we identified known (VEGF, AKT and mTOR) and novel (Wnt–β-catenin and Eph–ephrin) cell signaling pathways that are activated in NPC spheroids. Targeting these pathways in 3D spheroids using FDA-approved drugs was effective in monoculture and co-culture. These findings provide the first demonstration of the establishment of EBV-positive and EBV-negative NPC 3D spheroids with features that resemble advanced and metastatic NPCs. Furthermore, we show that NPC spheroids have potential use in identifying new drug targets.
Cardiorespiratory Fitness, Fat Mass, and Cardiometabolic Health with Endothelial Function, Arterial Elasticity, and Stiffness
Authors: AGBAJE ANDREW O. , BARKER ALAN R. , TUOMAINEN TOMI-PEKKA
Published: August 2021

Purpose

This study aimed to determine whether estimated cardiorespiratory fitness (CRF), fat mass (FM), lean mass (LM), and adiponectin bidirectionally associate with arterial function and structure and if CRF mediates the relationship between cardiometabolic health and arterial outcomes in 9- to 11-yr-old children drawn from the Avon Longitudinal Study of Parents and Children birth cohort, United Kingdom.

Methods

Brachial artery flow-mediated dilation (FMD), distensibility coefficient (DC), and carotid–radial pulse wave velocity (PWV) were measured by ultrasonography; CRF was measured during the submaximal ergometer test; total FM, trunk FM, and LM were measured by dual-energy x-ray absorptiometry; plasma adiponectin was measured by enzyme assay; and cardiometabolic health was computed based on the International Diabetes Federation criteria for diagnosing metabolic syndrome. We tested bidirectionality by including CRF, FM, LM, and adiponectin as exposures and FMD, DC, and PWV as outcomes, alternatively.

Results

Among 5566 participants (2816 (51%) girls; median age, 9.75 yr), CRF per body mass0.21 was directly related to DC (β (95% confidence interval) = 0.004 (<0.0001 to 0.008); P = 0.046), whereas CRF per LM0.54 was inversely associated with PWV (−0.034 (−0.063 to −0.003); 0.032) after adjusting for covariates. These associations remained in bidirectional analyses. Total FM, trunk FM, and LM were bidirectionally and positively associated with FMD and DC. Total FM and trunk FM but not LM had bidirectional and inverse associations with PWV. Adiponectin was not related to FMD, DC, or PWV. CRF partially mediated the associations of cardiometabolic health with FMD (1.5% mediation), DC (12.1% mediation), and PWV (3.5% mediation).

Conclusions

Associations of poor cardiometabolic health with adverse arterial structure and function in childhood may be mitigated by increasing CRF. Higher CRF was associated with better arterial structure whereas higher total FM and trunk FM were associated with better arterial function and structure. In the reverse analysis, healthy arterial structure and function were independently associated with increased total FM and trunk FM, suggesting an “arterial paradox.”
Physical Activity and Total Daily Energy Expenditure in Older US Adults: Constrained versus Additive Models
Authors: WILLIS ERIK A. , CREASY SETH A. , SAINT-MAURICE PEDRO F. , KEADLE SARAH KOZEY , PONTZER HERMANN , SCHOELLER DALE , TROIANO RICHARD P. , MATTHEWS CHARLES E.
Published: August 2021

Purpose

This study aimed to examine the shape of the relationship between physical activity (PA) and total energy expenditure (TEE) and to explore the role of energy balance status (negative, stable, positive) in influencing this association.

Methods

Cross-sectional. Participants were 584 older adults (50–74 yr) participating in the Interactive Diet and Activity Tracking in AARP study. TEE was assessed by doubly labeled water and PA by accelerometer. The relationship between PA and TEE was assessed visually and using nonlinear methods (restricted cubic splines). Percent weight change (>3%) over a 6-month period was used as a proxy measurement of energy balance status.

Results

TEE generally increased with increasing deciles of PA averaging 2354 (SD, 351) kcal·d−1 in the bottom decile to 2693 (SD, 480) kcal·d−1 in the top decile. Cubic spline models showed an approximate linear association between PA and TEE (linear relation, P < 0.0001; curvature, P = 0.920). Results were similar in subgroup analyses for individuals classified as stable or positive energy balance. For those in negative energy balance, TEE was generally flat with increasing deciles of PA averaging 2428 (SD, 285) kcal·d−1 in the bottom decile to 2372 (SD, 560) kcal·d−1 in the top decile.

Conclusions

Energy balance status seems to play an important role in the relationship between PA and TEE. When in a positive energy balance, the relationship between TEE and PA was consistent with an additive model; however, when energy balance was negative, TEE seems to be consistent with a constrained model. These findings support PA for weight gain prevention by increasing TEE; however, the effect of PA on TEE during periods of weight loss may be limited. An adequately powered, prospective study is warranted to confirm these exploratory findings.
Acute Cardiopulmonary and Muscle Oxygenation Responses to Normocapnic Hyperpnea Exercise in COPD
Authors: OUESLATI FERID , SAEY DIDIER , VÉZINA FÉLIX-ANTOINE , NADREAU ÉRIC , MARTIN MICKAËL , MALTAIS FRANÇOIS
Published: August 2021

Purpose

This study aimed to investigate cardiorespiratory responses and intercostal muscle oxygenation during normocapnic hyperpnea exercise in chronic obstructive pulmonary disease (COPD).

Methods

Twenty-two patients with COPD performed a cardiopulmonary cycling exercise test to assess peak oxygen consumption (V˙O2peak) and minute ventilation (V˙
Epeak). They also performed a normocapnic hyperpnea exercise alone, at 50%–60% of V˙
Epeak to exhaustion, using a respiratory device (Spirotiger) connected to a gas analyzer to monitor V˙O2, V˙
E, and end-tidal CO2 partial pressure. Cardiac output, and intercostal and vastus lateralis muscle oxygenation were continuously measured during exercise using finger photoplethysmography and near-infrared spectroscopy, respectively. Arterial blood gases (arterial PCO2) and inspiratory capacity were obtained at rest and at the end of hyperpnea exercise.

Results

The hyperpnea exercise lasted 576 ± 277 s at a V˙
E of 34.5 ± 12.1 L·min−1 (58% ± 6% of V˙
Epeak), a respiratory rate of 22 ± 4 breaths per minute, and a tidal volume of 1.43 ± 0.43 L. From rest to the end of hyperpnea exercise, V˙O2 increased by 0.35 ± 0.16 L·min−1 (P < 0.001), whereas end-tidal CO2 partial pressure and arterial PCO2 decreased by ~2 mm Hg (P = 0.031) and ~5 mm Hg (P = 0.002, n = 13), respectively. Moreover, inspiratory capacity fell from 2.44 ± 0.84 L at rest to 1.96 ± 0.59 L (P = 0.002). During the same period, heart rate and cardiac output increased from 69 ± 12 bpm and 4.94 ± 1.15 L·min−1 at rest to 87 ± 17 bpm (P = 0.002) and 5.92 ± 1.58 L·min−1 (P = 0.007), respectively. During hyperpnea exercise, intercostal deoxyhemoglobin and total hemoglobin increased by 14.26% ± 13.72% (P = 0.001) and 8.69% ± 12.49% (P = 0.003) compared with their resting value. However, during the same period, vastus lateralis oxygenation remained stable (P > 0.05).

Conclusions

In patients with COPD, normocapnic hyperpnea exercise provided a potent cardiorespiratory physiological stimulus, including dynamic hyperinflation, and increased intercostal deoxyhemoglobin consistent with enhanced requirement for muscle O2 extraction.
Membrane Proteins Increase with the Repeated Bout Effect
Authors: SIDKY SYLVIA R. , INGALLS CHRISTOPHER P. , LOWE DAWN A. , BAUMANN CORY W.
Published: August 2021

Purpose

The ability of skeletal muscle to adapt to eccentric (ECC) contraction-induced injury is known as the repeated bout effect (RBE). Despite the RBE being a well-established phenomenon observed in skeletal muscle, cellular and molecular events particularly those at the membranes that contribute to the adaptive potential of muscle have yet to be established. Therefore, the purpose of this study was to examine how membrane-associated proteins respond to the RBE.

Methods

Anterior crural muscles of C57BL/6 female mice (3–5 months) were subjected to repeated bouts of in vivo ECCs, with isometric torque being measured immediately before and after injury. A total of six bouts were completed with 7 d between each bout. Protein content of dystrophin, β-sarcoglycan, and junctophilin were then assessed via immunoblotting in injured and uninjured muscles.

Results

When expressed relative to preinjury isometric torque of bout 1, deficits in postinjury isometric torque during bout 2 (38%) did not differ from bout 1 (36%; P = 0.646) and were attenuated during bouts 3 through 6 (range, 24%–15%; P ≤ 0.014). Contents of dystrophin, β-sarcoglycan, and junctophilin did not change immediately after a single bout of 50 maximal ECCs (P ≥ 0.155); however, as a result of repeated bouts, contents of dystrophin, β-sarcoglycan, and junctophilin all increased compared with muscles that completed one or no bouts of ECC contractions (P ≤ 0.003).

Conclusions

The RBE represents a physiological measure of skeletal muscle plasticity. Here, we demonstrate that repeated bouts of ECC contractions increase contents of dystrophin, β-sarcoglycan, and junctophilin and attenuate postinjury torque deficits. Given our results, accumulation of membrane-associated proteins likely contributes to strength adaptations observed after repeated bouts of ECC contractions.