Additional Members Research Articles

Computational Stabilization of a Non‐heme Iron Enzyme Enables Efficient Evolution of New Function

Deep learning tools for enzyme design are rapidly emerging, and there is a critical need to evaluate their effectiveness in engineering workflows. Here we show that the deep learning‐based tool ProteinMPNN can be used to redesign Fe(II)/αKG superfamily enzymes for greater stability, solubility, and expression while retaining both native activity and industrially‐relevant non‐native functions. This superfamily has diverse catalytic functions and could provide a rich new source of biocatalysts for synthesis and industrial processes. Through systematic comparisons of directed evolution trajectories for a non‐native, remote C(sp3)‐H hydroxylation reaction, we demonstrate that the stabilized redesign can be evolved more efficiently than the wild‐type enzyme. After three rounds of directed evolution, we obtained a 6‐fold activity increase from the wild‐type parent and an 80‐fold increase from the stabilized variant. To generate the initial stabilized variant, we identified multiple structural and sequence constraints to preserve catalytic function. We applied these criteria to produce stabilized, catalytically active variants of a second Fe(II)/αKG enzyme, suggesting that the approach is generalizable to additional members of the Fe(II)/αKG superfamily. ProteinMPNN is user‐friendly and widely‐accessible, and our results provide a framework for the routine implementation of deep learning‐based protein stabilization tools in directed evolution workflows for novel biocatalysts.

Computational Stabilization of a Non-heme Iron Enzyme Enables Efficient Evolution of New Function.

Deep learning tools for enzyme design are rapidly emerging, and there is a critical need to evaluate their effectiveness in engineering workflows. Here we show that the deep learning-based tool ProteinMPNN can be used to redesign Fe(II)/αKG superfamily enzymes for greater stability, solubility, and expression while retaining both native activity and industrially-relevant non-native functions. This superfamily has diverse catalytic functions and could provide a rich new source of biocatalysts for synthesis and industrial processes. Through systematic comparisons of directed evolution trajectories for a non-native, remote C(sp3)-H hydroxylation reaction, we demonstrate that the stabilized redesign can be evolved more efficiently than the wild-type enzyme. After three rounds of directed evolution, we obtained a 6-fold activity increase from the wild-type parent and an 80-fold increase from the stabilized variant. To generate the initial stabilized variant, we identified multiple structural and sequence constraints to preserve catalytic function. We applied these criteria to produce stabilized, catalytically active variants of a second Fe(II)/αKG enzyme, suggesting that the approach is generalizable to additional members of the Fe(II)/αKG superfamily. ProteinMPNN is user-friendly and widely-accessible, and our results provide a framework for the routine implementation of deep learning-based protein stabilization tools in directed evolution workflows for novel biocatalysts.

Determinants of Willingness to Pay for Community-Based Health Insurance in Ethiopia: National Household Survey

The existing evidence on households' participation in Ethiopia's community-based health insurance (CBHI) scheme is limited, lacks representativeness, and lacks disaggregation. Thus, this study aims to assess households' willingness to pay (WTP) for CBHI membership and identify the factors that influence their decision to enroll. From February to May 2020, a nationwide cross-sectional household survey was conducted involving CBHI member and non-member households. The primary theories that informed this study were utility theory, social capital theory, and the health belief model. Sample included 5,976 households from 166 EAs—118 in CBHI regions and 48 in non-CBHI regions. A two-stage stratified cluster sampling used to select enumeration areas (EAs) and households from within these areas. The maximum amount of money at which participants responded "yes" in the bidding game exercise was used to measure WTP. Data were analyzed using STATA Version 16. Household expenditure was adjusted for key factors. Analyses explored WTP for CBHI by region, membership, and livelihood, with WTP based on mean maximum values. Linear regression identified influencing factors. The findings reveal that 30.9% of participants were active CBHI members. The average WTP increased with each additional family member, both in rural (AMD = 9.3 [6.8, 11.9]) and urban areas (AMD = 7.2 [1.0, 13.4]). In urban areas, WTP was also positively associated with the ability to pay (ATP) for CBHI (AMD = 64.1 [6.3, 121.8]). Male respondents and those who were literate in urban areas had higher mean WTP values of AMD = 39.8 [13.1, 66.4] and AMD = 56.8 [26.1, 87.4], respectively. Additionally, holding leadership positions in health and women's development initiatives positively influenced WTP. The study found that 30.9% of participants were active CBHI members. WTP for CBHI was higher among larger families. In urban areas, WTP was positively associated with the ability to pay (ATP) for CBHI, particularly among male and literate respondents. Community leadership roles in health and women's development also positively influenced WTP. To improve the CBHI scheme, adjustments should be made to premiums based on households' ability to pay. Community engagement should be strengthened, service quality enhanced, regional disparities addressed, and performance regularly monitored.

Addressing diabetes by elevating access to nutrition (ADELANTE) - A multi-level approach for improving household food insecurity and glycemic control among Latinos with diabetes: A randomized controlled trial

BackgroundLatinx adults are disproportionately impacted by the interrelated challenges of food insecurity and nutrition sensitive chronic diseases. Food and nutrition insecurity can exacerbate the development and progression of chronic diseases, such as diabetes. Sustainable, effective interventions aimed at improving food insecurity and diabetes management for Latinx populations are needed. MethodsThis hybrid type 1 trial evaluates the effectiveness of a multi-level intervention that includes a medically supportive food and behavioral lifestyle program on the primary outcome of Hemoglobin A1c (HbA1c) at 6 months. Latinx adults (n = 355) with type 2 diabetes (HbA1c of 6.0–12.0 %), overweight/obesity (BMI > 25 kg/m2), and self-reported risk of food insecurity will be randomized 1:1 to intervention (12 weekly deliveries of vegetables, fruits, and whole-grain foods + culturally-modified behavioral lifestyle program) versus control (food deliveries after a 6-month delay). Outcome asessments will occur at 0, 6 and 12 months, and include HbA1c, dietary intake, psychosocial health outcomes, and diabetes-related stressors. In addition, food insecurity and the impact of the intervention on up to two household members will be measured. Qualitative interviews with patients, healthcare providers, and community partners will be conducted in accordance with Reach, Effectivenes, Adoption, Implementation, and Maintenence (RE-AIM) framework to identify barriers and best practices for future dissemination. ConclusionsThe ADELANTE trial will provide novel insight to the effectiveness of a multi-level intervention on diabetes-related outcomes in Latinx adults. The mixed-method approach will also identity the reach of this ‘Food is Medicine’ intervention on additional household members to inform diabetes prevention efforts.Clinical Trial Registration: NCT05228860

Estimating the economic impact of blister-packaging on medication adherence and health care costs for a Medicare Advantage health plan.

Medication nonadherence is a persistent challenge in the United States, leading to increased health care resource utilization (HCRU) and health care costs and worsened health outcomes. Medicare Star Ratings is a program developed by the Centers for Medicare and Medicaid Services (CMS) to evaluate Medicare health plan quality and performance. Three of the Medicare Part D Star Ratings quality measures assess medication adherence, showing the importance CMS places on improving medication adherence in older adults. Although a variety of medication adherence-enhancing interventions are available to help promote adherence among patients, one intervention that has shown success historically is blister-packaging. To model the potential impact of blister-packaging chronic medications on HCRU and health care costs in the Medicare population. An economic model was developed to assess the potential impact of blister-packaging the 3 Medicare Star Ratings adherence measure medication classes: renin-angiotensin system antagonists (RASAs), statins, and noninsulin antidiabetics. The model perspective was that of a hypothetical Medicare Advantage health plan with a plan size of 100,000 members. A 12-month time horizon was used in the model. The dichotomous adherence threshold in the model was set at 80% or greater of the proportion of days covered (PDC). Literature-based references were used to inform both the impact of blister-packaging on the number of patients who become adherent as well as the impact of medication adherence on HCRU and health care costs for each of the medication classes. One-way sensitivity analyses and several scenario analyses were conducted to assess model uncertainty. Owing to increased adherence from the blister-packaging intervention, the hypothetical health plan in the analysis saw 776 additional members adherent to RASAs, 1,651 additional members adherent to statins, and 414 additional members adherent to oral antidiabetics. Although medication expenditure increased for all 3 medication classes (RASAs: $274,963; statins: $730,083; oral antidiabetics: $100,529), medical costs decreased across all classes (RASAs: -$4,098,848; statins: -$5,549,699; oral antidiabetics: -$917,968). Total net health care costs decreased by $3,823,885 for RASAs (-$3.19 per member per month [PMPM]), $4,819,616 for statins (-$4.02 PMPM), and $817,438 for oral antidiabetics (-$0.68 PMPM). The entire Medicare Advantage population scenario analysis saw reductions in total health care costs of $1,081,394,737 for RASAs, $1,362,987,376 for statins, and $231,171,496 for oral antidiabetics. Dispensing chronic medications with blister-packaging for Medicare Advantage health plan patients was modeled to reduce HCRU and health care costs. Future studies are needed to assess whether the impact of blister-packaging medications is tied to reductions in HCRU and health care costs in real-world settings.

The Nuclear NF-κB Regulator IκBζ: Updates on Its Molecular Functions and Pathophysiological Roles.

More than a decade after the discovery of the classical cytoplasmic IκB proteins, IκBζ was identified as an additional member of the IκB family. Unlike cytoplasmic IκB proteins, IκBζ has distinct features, including its nuclear localization, preferential binding to NF-κB subunits, unique expression properties, and specialized role in NF-κB regulation. While the activation of NF-κB is primarily controlled by cytoplasmic IκB members at the level of nuclear entry, IκBζ provides an additional layer of NF-κB regulation in the nucleus, enabling selective gene activation. Human genome-wide association studies (GWAS) and gene knockout experiments in mice have elucidated the physiological and pathological roles of IκBζ. Despite the initial focus to its role in activated macrophages, IκBζ has since been recognized as a key player in the IL-17-triggered production of immune molecules in epithelial cells, which has garnered significant clinical interest. Recent research has also unveiled a novel molecular function of IκBζ, linking NF-κB and the POU transcription factors through its N-terminal region, whose role had remained elusive for many years.

Impact of Forced Relocation and Community Integration on Household Well-being of Urban Refugees in Kenya

Forced displacement is a global issue that affects millions of people all over the world. Refugees are forced to flee their homes due to conflict, persecution, or other forms of violence. The majority of refugees often end up in urban areas, where they face numerous challenges, including social exclusion, limited access to basic services, and economic marginalization. This study examines the relationship between forced displacement, social cohesion, and household welfare outcomes of urban refugees in Kenya. The study utilized household data on urban refugees in Kenya sourced from the United Nations High Commission for Refugees (UNHCR), 2020 [85], and, the Kenya Continuous Household Survey (KCHS), 2019 [41]. The study estimates were regressed using the Ordinary Least Squares method and diagnostic tests were done to determine the accuracy of the estimates. The study found that an additional member in an urban refugee household led to an asset increment of 6.7%. The study further found that an additional habitable dwelling in an urban refugee household increased family assets by 71.9%. Utilization of mobile banking services, having nuclear members abroad, and participation in community-based organizations also had a positive impact on family asset accumulation in a refugee household. The study also found that utilization of mobile money transfer (M-PESA), negatively influenced family assets. This study contributes to the literature on Urban refugees as it gives insights into factors that can improve welfare and enhance social cohesion between refugees and host communities in Kenya. Gaps have also been identified in the mandate of UNHCR beyond the camps and the role of the Kenyan Government in urban housing programs for refugees.

Enterobacter adelaidei sp. nov. Isolation of an extensively drug resistant strain from hospital wastewater in Australia and the global distribution of the species

BackgroundEnterobacter species are included among the normal human gut microflora and persist in a diverse range of other environmental niches. They have become important opportunistic nosocomial pathogens known to harbour plasmid-mediated multi-class antimicrobial resistance (AMR) determinants. Global AMR surveillance of Enterobacterales isolates shows the genus is second to Klebsiella in terms of frequency of carbapenem resistance. Enterobacter taxonomy is confusing and standard species identification methods are largely inaccurate or insufficient. There are currently 27 named species and a total of 46 taxa in the genus distinguishable via average nucleotide identity (ANI) calculation between pairs of genomic sequences. Here we describe an Enterobacter strain, ECC3473, isolated from the wastewater of an Australian hospital whose species could not be determined by standard methods nor by ribosomal RNA gene multi-locus typing. AimTo characterise ECC3473 in terms of phenotypic and genotypic antimicrobial resistance, biochemical characteristics and taxonomy as well as to determine the global distribution of the novel species to which it belongs. MethodsStandard broth dilution and disk diffusion were used to determine phenotypic AMR. The strain’s complete genome, including plasmids, was obtained following long- and short read sequencing and a novel long/short read hybrid assembly and polishing, and the genomic basis of AMR was determined. Phylogenomic analysis and quantitative measures of relatedness (ANI, digital DNA-DNA hybridisation, and difference in G+C content) were used to study the taxonomic relationship between ECC3473 and Enterobacter type-strains. NCBI and PubMLST databases and the literature were searched for additional members of the novel species to determine its global distribution. ResultsECC3473 is one of 21 strains isolated globally belonging to a novel Enterobacter species for which the name, Enterobacter adelaidei sp. nov. is proposed. The novel species was found to be resilient in its capacity to persist in contaminated water and adaptable in its ability to accumulate multiple transmissible AMR determinants. ConclusionE. adelaidei sp. nov. may become increasingly important to the dissemination of AMR.

In-situ observations of resident space objects with the CHEOPS space telescope

The CHaracterising ExOPlanet Satellite (CHEOPS) is a partnership between the European Space Agency and Switzerland with important contributions by 10 additional ESA member States. It is the first S-class mission in the ESA Science Programme. CHEOPS has been flying on a Sun-synchronous low Earth orbit since December 2019, collecting millions of short-exposure images in the visible domain to study exoplanet properties.A small yet increasing fraction of CHEOPS images show linear trails caused by resident space objects crossing the instrument field of view. CHEOPS’ orbit is indeed particularly favourable to serendipitously detect objects in its vicinity as the spacecraft rarely enters the Earth's shadow, sits at an altitude of 700 km, and observes with moderate phase angles relative to the Sun. This observing configuration is quite powerful, and it is complementary to optical observations from the ground.To characterize the population of satellites and orbital debris observed by CHEOPS, all and every science images acquired over the past 3 years have been scanned with a Hough transform algorithm to identify the characteristic linear features that these objects cause on the images. Thousands of trails have been detected. This statistically significant sample shows interesting trends and features such as an increased occurrence rate over the past years as well as the fingerprint of the Starlink constellation. The cross-matching of individual trails with catalogued objects is underway as we aim to measure their distance at the time of observation and deduce the apparent magnitude of the detected objects.As space agencies and private companies are developing new space-based surveillance and tracking activities to catalogue and characterize the distribution of small debris, the CHEOPS experience is timely and relevant. With the first CHEOPS mission extension currently running until the end of 2026, and a possible second extension until the end of 2029, the longer time coverage will make our dataset even more valuable to the community, especially for characterizing objects with recurrent crossings.

Intra-strain colony biofilm heterogeneity in uropathogenic Escherichia coli and the effect of the NlpI lipoprotein

Biofilm growth facilitates the interaction of uropathogenic Escherichia coli (UPEC) with the host environment. The extracellular polymeric substances (EPS) of UPEC biofilms are composed prominently of curli amyloid fiber and cellulose polysaccharide. When the organism is propagated as a colony biofilm on agar media, these macromolecules can generate pronounced macroscopic structures. Moreover, curli/cellulose associate tightly with Congo red, generating a characteristic pink-to-red staining pattern when the media is supplemented with this dye. Among different clinical isolates of UPEC, changes in the abundance of curli/cellulose can lead to diverse colony biofilm phenotypes on a strain-by-strain basis. Nevertheless, for any given isolate, these phenotypes are classically homogenous throughout the colony biofilm. Here, we report that a subset of clinical UPEC isolates display heterogenous ‘peppermint’ colony biofilms, with distinct pale and red subpopulations. Through isolation of these subpopulations and whole genome sequencing, we demonstrate various emergent mutations associated with the phenomenon, including within the gene encoding the outer membrane lipoprotein nlpI. Deletion of nlpI within independent strain-backgrounds increased biofilm rugosity, while its overexpression induced the peppermint phenotype. Upregulation of EPS-associated proteins and transcripts was likewise observed in the absence of nlpI. Overall, these results demonstrate that EPS elaboration in UPEC is impacted by nlpI. More broadly, this phenomenon of intra-strain colony biofilm heterogeneity may be leveraged as a tool to identify additional members within the broad collection of genes that regulate or otherwise affect biofilm formation.

Ideas For Estate and Transition Planning: Gifting Ownership

For many farm and ranch families, bringing children or grandchildren into the operation is the ultimate goal. Successfully bringing additional family members into the operation may require some creativity as all parties need to maintain a viable standard of living. This article is part of a series that will highlight ideas and tactics for bringing another family member into the operation.

Efektivitas Ketentuan Insentif dan Disinsentif Terkait Perizinan Pemanfaatan Ruang Pada Zona Perdagangan dan Jasa Berdasarkan Perda Nomor 5 Tahun 2012 Tentang Rencana Tata Ruang Wilayah Kota Sungai Penuh Tahun 2011 – 2031

Even though the Regional Regulation on the Regional RTRW of Sungai Banyak City has been stipulated, the use of existing space is not yet in accordance with the plans that have been determined. This is indicated by the fact that the building does not meet the building construction requirements for both KDB, GSB and KDH. Provisions on Incentives and Disincentives Related to Licensing for Space Utilization in Trade and Service Zones based on the Regional Regulation on the RTRW of Sungai Banyak City have not been implemented effectively, this can be seen from the 182 Building Approvals that have been issued by the Sungai Full City Government from 2021 to 2023, they are not implemented provisions for incentives and disincentives for controlling space utilization. Factors that become obstacles include the fact that the Regional Regulation has not yet been established, there is still a lack of Satpol PP members, the facilities and infrastructure are still not complete, the community's legal awareness is still low, and people are building houses using the process of adjunct direction from traditional institutions. The efforts made by the Regional Government of Sungai Banyak City to overcome these obstacles include drafting the Draft Regional Regulation on the RTRW Revision, proposing additional members of the Civil Service Police Unit, maximizing honorary personnel, procuring facilities and infrastructure, socializing the Regional Regulation on Spatial Planning to the community, collaborating with traditional institutions in the planning process , utilization and control of the spatial planning of Sungai Banyak City.

Exome sequencing in undiagnosed congenital myopathy reveals new genes and refines genes–phenotypes correlations

BackgroundCongenital myopathies are severe genetic diseases with a strong impact on patient autonomy and often on survival. A large number of patients do not have a genetic diagnosis, precluding genetic counseling and appropriate clinical management. Our objective was to find novel pathogenic variants and genes associated with congenital myopathies and to decrease diagnostic odysseys and dead-end.MethodsTo identify pathogenic variants and genes implicated in congenital myopathies, we established and conducted the MYOCAPTURE project from 2009 to 2018 to perform exome sequencing in a large cohort of 310 families partially excluded for the main known genes.ResultsPathogenic variants were identified in 156 families (50%), among which 123 families (40%) had a conclusive diagnosis. Only 44 (36%) of the resolved cases were linked to a known myopathy gene with the corresponding phenotype, while 55 (44%) were linked to pathogenic variants in a known myopathy gene with atypical signs, highlighting that most genetic diagnosis could not be anticipated based on clinical–histological assessments in this cohort. An important phenotypic and genetic heterogeneity was observed for the different genes and for the different congenital myopathy subtypes, respectively. In addition, we identified 14 new myopathy genes not previously associated with muscle diseases (20% of all diagnosed cases) that we previously reported in the literature, revealing novel pathomechanisms and potential therapeutic targets.ConclusionsOverall, this approach illustrates the importance of massive parallel gene sequencing as a comprehensive tool for establishing a molecular diagnosis for families with congenital myopathies. It also emphasizes the contribution of clinical data, histological findings on muscle biopsies, and the availability of DNA samples from additional family members to the diagnostic success rate. This study facilitated and accelerated the genetic diagnosis of congenital myopathies, improved health care for several patients, and opened novel perspectives for either repurposing of existing molecules or the development of novel treatments.

Improving ensemble data assimilation through Probit-space Ensemble Size Expansion for Gaussian Copulas (PESE-GC)

Abstract. Small forecast ensemble sizes (< 100) are common in the ensemble data assimilation (EnsDA) component of geophysical forecast systems, thus limiting the error-constraining power of EnsDA. This study proposes an efficient and embarrassingly parallel method to generate additional ensemble members: the Probit-space Ensemble Size Expansion for Gaussian Copulas (PESE-GC; “peace gee see”). Such members are called “virtual members”. PESE-GC utilizes the users' knowledge of the marginal distributions of forecast model variables. Virtual members can be generated from any (potentially non-Gaussian) multivariate forecast distribution that has a Gaussian copula. PESE-GC's impact on EnsDA is evaluated using the 40-variable Lorenz 1996 model, several EnsDA algorithms, several observation operators, a range of EnsDA cycling intervals, and a range of forecast ensemble sizes. Significant improvements to EnsDA (p<0.01) are observed when either (1) the forecast ensemble size is small (≤20 members), (2) the user selects marginal distributions that improve the forecast model variable statistics, and/or (3) the rank histogram filter is used with non-parametric priors in high-forecast-spread situations. These results motivate development and testing of PESE-GC for EnsDA with high-order geophysical models.

Sizing distributed energy resources in a renewable energy community with a grid-aware internal market structure

This paper proposes a cooperative approach aimed at distributed energy resources sizing in a renewable energy community, with considerations of the community’s optimal operation, impact on the electrical grid and an allocation of the benefits to its members. To this purpose, multiple investment modes are evaluated via a two-step procedure. In the first step, the size of renewable energy sources is determined by solving an optimization problem that maximizes community welfare, considering network and investments. In the second step, an optimization problem maximizing additional community member profit with price regularization is solved. This step shares benefits among community members. The potential of the proposed procedure is illustrated using a benchmark Dickert-LV network. This is a full y cooperative framework where the community operator is ensuring adequate grid operation, operational planning and sizing of new investments.

Pelaksanaan Mekanisme Program Keluarga Harapan (PKH) di Kota Medan

The PKH mechanism is also regulated based on Minister of Social Affairs Regulation No. Minister of Social Affairs Regulation no. 1 of 2018 concerning the Family Hope Program, which includes planning, determining prospective PKH participants, validating data on potential PKH beneficiary recipients, determining PKH beneficiary families, distributing PKH social assistance, PKH assistance, increasing family capacity, verifying PKH KPM commitments, updating data KPM PKH, transformation of PKH participation. The type of research used is descriptive with a qualitative approach. The data sources in this research are primary data and secondary data. The research informants were the Head of the Medan City Social Service, Secretary of the Social Service, Coordinator of the Family Hope Program Implementation Unit, beneficiary family recipients, and Head of the KPM Group. Data collection techniques can be done in several ways, namely interviews, observation and documentation. Data analysis techniques: data collection stage, data reduction stage, data presentation stage and conclusion drawing stage. Research results: In general, the PKH implementation mechanism is based on Article 32 of the Ministry of Social Affairs Regulation No. 1 of 2018, there are 10 stages in the PKH implementation mechanism, all the main processes in implementing PKH activities always involve the Social Services and PKH Facilitators and are in accordance with the PKH implementation guidelines, but have not been implemented optimally as per the stage mechanism that has been regulated. This can be seen starting from the planning process which is hampered by data proposing additional PKH beneficiary members being far from what was planned, so that it does not cover all the poor people in Medan City who receive social assistance from the Family Hope Program, the time for validating data on potential recipients is relatively short, resulting in the determination of PKH beneficiary families that are not on target.

CBP60b clade proteins are prototypical transcription factors mediating immunity.

Transcriptional reprogramming is critical for plant immunity. Several calmodulin (CaM)-binding protein 60 (CBP60) family transcription factors (TFs) in Arabidopsis (Arabidopsis thaliana), including CBP60g, Systemic Acquired Resistance Deficient 1 (SARD1), CBP60a, and CBP60b, are critical for and show distinct roles in immunity. However, there are additional CBP60 members whose function is unclear. We report here that Arabidopsis CBP60c-f, four uncharacterized CBP60 members, play redundant roles with CBP60b in the transcriptional regulation of immunity responses, whose pCBP60b-driven expression compensates the loss of CBP60b. By contrast, neither CBP60g nor SARD1 is inter-changeable with CBP60b, suggesting clade-specific functionalization. We further show that function of CBP60b clade TFs relies on DNA-binding domains (DBDs) and CaM-binding domains, suggesting that they are downstream components of calcium signaling. Importantly, we demonstrate that CBP60s encoded in earliest land plant lineage Physcomitrium patens and Selaginella moellendorffii, are functionally homologous to Arabidopsis CBP60b, suggesting that the CBP60b clade contains the prototype TFs of the CBP60 family. Furthermore, tomato and cucumber CBP60b-like genes rescue the defects of Arabidopsis cbp60b and activate the expression of tomato and cucumber SALICYLIC ACID INDUCTION DEFICIIENT2 (SID2) and ENHANCED DISEASE SUSCEPTIBILITY 1 (EDS1) genes, suggesting that immune response pathways centered on CBP60b are also evolutionarily conserved. Together, these findings suggest CBP60b clade transcription factors are functionally conserved in evolution and positively mediate immunity.

Spectroscopic Confirmation of an Ultra-Massive Galaxy in a Protocluster at z ∼ 4.9

We present spectroscopic confirmation of an ultra-massive galaxy (UMG) with log(M⋆/M⊙)=10.98±0.07 at zspec=4.8947 in the Extended Groth Strip (EGS), based on deep observations of Ly α emission with Keck/DEIMOS. The ultra-massive galaxy (UMG-28740) is the most massive member in one of the most significant overdensities in the EGS, with four additional photometric members with log(M⋆/M⊙)>10.5 within Rproj∼1 cMpc. Spectral energy distribution (SED) fitting using a large suite of star formation histories and two sets of high-quality photometry from ground- and space-based facilities consistently estimates the mass of this object to be log(M⋆/M⊙)∼11 with a small standard deviation between measurements ( σ=0.07). While the best-fit SED models agree on stellar mass, we find discrepancies in the estimated star formation rate for UMG-28740, resulting in either a star-forming or quiescent system. 𝐽𝑊𝑆𝑇/NIRCam photometry of UMG-28740 strongly favors a quiescent scenario, demonstrating the need for high-quality mid-IR observations. Assuming the galaxy to be quiescent, UMG-28740 formed the bulk of its stars at z>10 and is quenching at z∼8, resulting in a high star formation efficiency at high redshift ( ϵ∼0.2 at z∼5 and ϵ≳1 at z≳8). As the most massive galaxy in its protocluster environment, UMG-28740 is a unique example of the impossibly early galaxy problem.

Advisory Panel Guidance on Minimum Retesting Intervals for Lab Tests

What Is the Issue? Lab test overuse can contribute to further unnecessary follow-up and testing, negative patient experiences, potentially inappropriate treatments, and the inefficient use of health care resources. One review of lab testing in Canada found that around 22% of blood tests were likely unnecessary. One strategy to address lab test overuse is to establish minimal retesting intervals that can be implemented in medical laboratories to help identify and manage potentially inappropriate lab test requests. Minimum retesting intervals suggest the minimum time before a test should be repeated based on the biochemical properties of the test and the clinical situation in which it is used. They are intended to inform clinical decisions about repeat testing. The importance of lab resource stewardship is being addressed by Choosing Wisely Canada through Using Labs Wisely, a consortium of more than 150 hospitals committed to driving the appropriate use of lab testing in Canada. The hospitals participating in Using Labs Wisely identified a need for guidance on minimum retesting intervals for commonly used lab tests. What Did We Do? Choosing Wisely Canada and CADTH partnered to convene an independent time-limited advisory panel to develop consensus-based recommendations for minimum retesting intervals for 7 commonly used lab tests (antinuclear antibody [ANA], B-type natriuretic peptide [BNP] and N-terminal pro b-type natriuretic peptide [NT-proBNP], Hemoglobin A1C, lipase, lipid panel, serum protein electrophoresis [SPEP], and thyroid stimulating hormone [TSH]) in prespecified patient populations. The advisory panel included core and specialist members who were recruited from across Canada. The 7 core advisory panel members brought together expertise in laboratory medicine, family practice, and patient lived experience. Seven additional specialist members brought expertise in endocrinology, cardiology, pediatric cardiology, rheumatology, hematology oncology, gastroenterology, and general internal medicine. The Advisory Panel on Minimum Retesting Intervals considered patient group input, evidence from focused literature reviews, equity considerations, and clinical expertise. Through facilitated discussion, they reached consensus on the recommendations for minimum retesting intervals. Following external feedback, the recommendations for BNP and NT-proBNP and lipid panels were removed, and this document includes recommendations for minimum retesting intervals for 5 lab tests. These are not recommendations for repeat testing. They are recommendations that if testing is undertaken, it should not be repeated sooner than the indicated intervals. They are not intended to replace clinical judgment as there may be exceptions in which the recommendations do not apply. What Is the Potential Impact? The recommendations on minimum retesting intervals can support the hospitals participating in Choosing Wisely Canada’s Using Labs Wisely program in their effort to reduce unnecessary lab tests and their impact on patients, providers, health systems, and the environment. The recommendations may also be relevant to community and hospital lab stewardship efforts and may address the appropriate use of the 5 lab tests by enabling changes in lab test ordering in both inpatient and outpatient settings.

Myotonia congenita in a Greek cohort: Genotype spectrum and impact of the CLCN1:c.501C > G variant as a genetic modifier.

Myotonia congenita (MC) is the most common hereditary channelopathy in humans. Characterized by muscle stiffness, MC may be transmitted as either an autosomal dominant (Thomsen) or a recessive (Becker) disorder. MC is caused by variants in the voltage-gated chloride channel 1 (CLCN1) gene, important for the normal repolarization of the muscle action potential. More than 250 disease-causing variants in the CLCN1 gene have been reported. This study provides an MC genotype-phenotype spectrum in a large cohort of Greek patients and focuses on novel variants and disease epidemiology, including additional insights for the variant CLCN1:c.501C > G. Sanger sequencing for the entire coding region of the CLCN1 gene was performed. Targeted segregation analysis of likely candidate variants in additional family members was performed. Variant classification was based on American College of Medical Genetics (ACMG) guidelines. Sixty-one patients from 47 unrelated families were identified, consisting of 51 probands with Becker MC (84%) and 10 with Thomsen MC (16%). Among the different variants detected, 11 were novel and 16 were previously reported. The three most prevalent variants were c.501C > G, c.2680C > T, and c.1649C > G. Additionally, c.501C > G was detected in seven Becker cases in-cis with the c.1649C > G. The large number of patients in whom a diagnosis was established allowed the characterization of genotype-phenotype correlations with respect to both previously reported and novel findings. For the c.501C > G (p.Phe167Leu) variant a likely nonpathogenic property is suggested, as it only seems to act as an aggravating modifying factor in cases in which a pathogenic variant triggers phenotypic expression.