MDC1 Interacts with TOPBP1 to Maintain Chromosomal Stability during Mitosis.

In mitosis, cells inactivate DNA double-strand break (DSB) repair pathways to preserve genome stability. However, some early signaling events still occur, such as recruitment of the scaffold protein MDC1 to phosphorylated histone H2AX at DSBs. Yet, it remains unclear whether these events are important for maintaining genome stability during mitosis. Here, we identify a highly conserved protein-interaction surface in MDC1 that is phosphorylated by CK2 and recognized by the DNA-damage response mediator protein TOPBP1. Disruption of MDC1-TOPBP1 binding causes a specific loss of TOPBP1 recruitment to DSBs in mitotic but not interphase cells, accompanied by mitotic radiosensitivity, increased micronuclei, and chromosomal instability. Mechanistically, we find that TOPBP1 forms filamentous structures capable of bridging MDC1 foci in mitosis, indicating that MDC1-TOPBP1 complexes tether DSBs until repair is reactivated in the following G1 phase. Thus, we reveal an important, hitherto-unnoticed cooperation between MDC1 and TOPBP1 in maintaining genome stability during cell division.

Proteins required for stereocilia elongation during mammalian hair cell development ensure precise and steady heights during adult life.

The hair bundle, or stereocilia bundle, is the mechanosensory compartment of hair cells (HCs) in the inner ear. To date, most mechanistic studies have focused on stereocilia bundle morphogenesis, and it remains unclear how this organelle critical for hearing preserves its precise dimensions during life in mammals. The GPSM2-GNAI complex occupies the distal tip of stereocilia in the tallest row and is required for their elongation during development. Here, we ablate GPSM2-GNAI in adult mouse HCs after normal stereocilia elongation is completed. We observe a progressive height reduction of the tallest row stereocilia totaling ~600 nm after 12 wk in Gpsm2 mutant inner HCs. To measure GPSM2 longevity at tips, we generated a HaloTag-Gpsm2 mouse strain and performed pulse-chase experiments in vivo. Estimates using pulse-chase or tracking loss of GPSM2 immunolabeling following Gpsm2 inactivation suggest that GPSM2 is relatively long-lived at stereocilia tips with a half-life of 9 to 10 d. Height reduction coincides with dampened auditory brainstem responses evoked by low-frequency stimuli in particular. Finally, GPSM2 is required for normal tip enrichment of elongation complex (EC) partners MYO15A, WHRN, and EPS8, mirroring their established codependence during development. Taken together, our results show that the EC is also essential in mature HCs to ensure precise and stable stereocilia height and for sensitive detection of a full range of sound frequencies.

IMPACT OF REAL-LIFE ENVIRONMENTAL EXPOSURES ON REPRODUCTION: Evidence for reproductive health effects following exposure to hydraulic fracturing chemical mixtures.

In Brief: Unconventional oil and natural gas (UOG) operations, particularly hydraulic fracturing, have revolutionized oil and gas production, using and containing complex mixtures of chemicals that may impact reproductive health. While there is growing evidence for effects on births in hydraulic fracturing/UOG regions and good mechanistic evidence for potential reproductive toxicity, there is much research still needed to make firm conclusions about these practices and reproductive health. Abstract: Unconventional oil and natural gas (UOG) operations have emerged over the last four decades to transform oil and gas production in the United States and globally by unlocking previously inaccessible hydrocarbon deposits. UOG development utilizes many compounds associated with conventional oil and gas, as well as some specific to UOG extraction, particularly during hydraulic fracturing (HF). While research is increasing on UOG chemicals and their mixtures, this review discusses the current evidence for reproductive toxicity following exposures to UOG/HF mixtures. These complex chemical mixtures have been demonstrated to interact with numerous mechanisms known to influence reproductive health. A growing number of environmental and controlled laboratory testing studies have reported adverse reproductive health effects in animals exposed to various UOG chemical mixtures. An expanding body of epidemiological literature has assessed adverse birth outcomes, although none has directly examined reproductive measures such as time to pregnancy, semen quality, and other direct measures of fertility. The existing literature provides moderate evidence for decreased birth weights, increased risk of small for gestational age and/or preterm birth, increased congenital abnormalities, and increased infant mortality, though importantly, studies are widely variable in methods used. Most studies utilized distance from UOG operations as an exposure proxy and did not measure actual chemical exposures experienced by those living near these operations. As such, while there is growing evidence for effects on births in these regions and good mechanistic evidence for potential reproductive toxicity, there is much research still needed to make firm conclusions about UOG development and reproductive health.

Improved survival in COVID-19 related sepsis and ARDS treated with a unique "triple therapy" including therapeutic plasma exchange: A single center retrospective analysis.

BACKGROUND: Throughout the COVID-19 pandemic, the mortality of critically ill patients remained high. Our group developed a treatment regimen targeting sepsis and ARDS which we labeled "triple therapy" consisting of (1) corticosteroids, (2) therapeutic plasma exchange (TPE), and (3) timely intubation with lung protective ventilation. Our propensity analysis assesses the impact of triple therapy on survival in COVID-19 patients with sepsis and ARDS. METHODS: Retrospective propensity analysis comparing triple therapy to no triple therapy in adult critically ill COVID-19 patients admitted to the Intensive Care Unit at Lexington Medical Center from 1 March 2020 through 31 October 2021. RESULTS: Eight hundred and fifty-one patients were admitted with COVID-19 and 53 clinical and laboratory variables were analyzed. Multivariable analysis revealed that triple therapy was associated with increased survival (OR: 1.91; P = .008). Two propensity score-adjusted models demonstrated an increased likelihood of survival in patients receiving triple therapy. Patients with thrombocytopenia were among those most likely to experience increased survival if they received early triple therapy. Decreased survival was observed with endotracheal intubation ≥7 days from hospital admission (P < .001) and there was a trend toward decreased survival if TPE was initiated ≥6 days from hospital admission (P = .091). CONCLUSION: Our analysis shows that early triple therapy, defined as high-dose methylprednisolone, TPE, and timely invasive mechanical ventilation within the first 96 hours of admission, may improve survival in critically ill septic patients with ARDS secondary to COVID-19 infection. Further studies are needed to define specific phenotypes and characteristics that will identify those patients most likely to benefit.

Structure of the human RAD17-RFC clamp loader and 9-1-1 checkpoint clamp bound to a dsDNA-ssDNA junction.

The RAD9-RAD1-HUS1 (9-1-1) clamp forms one half of the DNA damage checkpoint system that signals the presence of substantial regions of single-stranded DNA arising from replication fork collapse or resection of DNA double strand breaks. Loaded at the 5'-recessed end of a dsDNA-ssDNA junction by the RAD17-RFC clamp loader complex, the phosphorylated C-terminal tail of the RAD9 subunit of 9-1-1 engages with the mediator scaffold TOPBP1 which in turn activates the ATR kinase, localised through the interaction of its constitutive partner ATRIP with RPA-coated ssDNA. Using cryogenic electron microscopy (cryoEM) we have determined the structure of a complex of the human RAD17-RFC clamp loader bound to human 9-1-1, engaged with a dsDNA-ssDNA junction. The structure answers the key questions of how RAD17 confers specificity for 9-1-1 over PCNA, and how the clamp loader specifically recognises the recessed 5' DNA end and fixes the orientation of 9-1-1 on the ssDNA.

Coherent Interactions between Silicon-Vacancy Centers in Diamond.

We report tunable excitation-induced dipole-dipole interactions between silicon-vacancy color centers in diamond at cryogenic temperatures. These interactions couple centers into collective states, and excitation-induced shifts tag the excitation level of these collective states against the background of excited single centers. By characterizing the phase and amplitude of the spectrally resolved interaction-induced signal, we observe oscillations in the interaction strength and population state of the collective states as a function of excitation pulse area. Our results demonstrate that excitation-induced dipole-dipole interactions between color centers provide a route to manipulating collective intercenter states in the context of a congested, inhomogeneous ensemble.

Early career researchers benefit from inclusive, diverse and international collaborations: Changing how academic institutions utilize the seminar series.

Efforts to make research environments more inclusive and diverse are beneficial for the next generation of Great Lakes researchers. The global COVID-19 pandemic introduced circumstances that forced graduate programs and academic institutions to re-evaluate and promptly pivot research traditions, such as weekly seminar series, which are critical training grounds and networking opportunities for early career researchers (ECRs). While several studies have established that academics with funded grants and robust networks were better able to weather the abrupt changes in research and closures of institutions, ECRs did not. In response, both existing and novel partnerships provided a resilient network to support ECRs at an essential stage of their career development. Considering these challenges, we sought to re-frame the seminar series as a virtual collaboration for ECRs. Two interdisciplinary graduate programs, located in different countries (Windsor, Canada, and Detroit, USA) invested in a year-long partnership to deliver a virtual-only seminar series that intentionally promoted: the co-creation of protocols and co-led roles, the amplification of justice, equity, diversity and inclusion throughout all aspects of organization and representation, engagement and amplification through social media, the integration of social, scientific and cultural research disciplines, all of which collectively showcased the capacity of our ECRs to lead, organize and communicate. This approach has great potential for application across different communities to learn through collaboration and sharing, and to empower the next generation to find new ways of working together.

Hidden Silicon-Vacancy Centers in Diamond.

We characterize a high-density sample of negatively charged silicon-vacancy (SiV^{-}) centers in diamond using collinear optical multidimensional coherent spectroscopy. By comparing the results of complementary signal detection schemes, we identify a hidden population of SiV^{-} centers that is not typically observed in photoluminescence and which exhibits significant spectral inhomogeneity and extended electronic T_{2} times. The phenomenon is likely caused by strain, indicating a potential mechanism for controlling electric coherence in color-center-based quantum devices.

Phosphorylation-dependent assembly and coordination of the DNA damage checkpoint apparatus by Rad4(TopBP1).

The BRCT-domain protein Rad4(TopBP1) facilitates activation of the DNA damage checkpoint in Schizosaccharomyces pombe by physically coupling the Rad9-Rad1-Hus1 clamp, the Rad3(ATR) -Rad26(ATRIP) kinase complex, and the Crb2(53BP1) mediator. We have now determined crystal structures of the BRCT repeats of Rad4(TopBP1), revealing a distinctive domain architecture, and characterized their phosphorylation-dependent interactions with Rad9 and Crb2(53BP1). We identify a cluster of phosphorylation sites in the N-terminal region of Crb2(53BP1) that mediate interaction with Rad4(TopBP1) and reveal a hierarchical phosphorylation mechanism in which phosphorylation of Crb2(53BP1) residues Thr215 and Thr235 promotes phosphorylation of the noncanonical Thr187 site by scaffolding cyclin-dependent kinase (CDK) recruitment. Finally, we show that the simultaneous interaction of a single Rad4(TopBP1) molecule with both Thr187 phosphorylation sites in a Crb2(53BP1) dimer is essential for establishing the DNA damage checkpoint.

Quantum-well laser diodes for frequency comb spectroscopy.

We demonstrate simple optical frequency combs based on semiconductor quantum well laser diodes. The frequency comb spectrum can be tailored by choice of material properties and quantum-well widths, providing spectral flexibility. We demonstrate the correlation in the phase fluctuations between two devices on the same chip by generating a radio-frequency dual comb spectrum.

The Role of the NICU in Father Involvement, Beliefs, and Confidence: A Follow-up Qualitative Study.

BACKGROUND: Fathers are important to infant outcomes. Infants of involved fathers have improved weight gain, sleep, and psychosocial behaviors. Father involvement with neonatal intensive care unit (NICU) infants reduces the length of stay. PURPOSE: The purpose of this study was to explore and describe involvement, confidence, and beliefs of fathers of infants who were hospitalized in the NICU and discharged home in order to begin to investigate NICU father involvement from a longitudinal perspective. METHODS: This exploratory qualitative study was conducted 4 to 5 years after the initial NICU stay using telephone interviews. Fathers who participated in this study were selected from participants of a previous NICU study. Qualitative analysis was conducted using standard procedures for grounded theory. RESULTS: Nineteen fathers participated in the study. The major themes were "It was scary," "Just be there," "It was rough," "It's not about yourself," "A special bond," and "Almost a treat." The fathers reported that the NICU providers, nurses, and staff helped them to overcome uncertainty and lack of knowledge, which helped them improve their confidence and involvement during the NICU stay. IMPLICATIONS FOR PRACTICE: Fathers see nurses as a source of support. Nurses can encourage fathers to visit regularly and participate in infant care activities. NICU presence aids fathers in developing confidence and knowledge in parenting during their child's infancy, which can set the stage for ongoing involvement. IMPLICATIONS FOR RESEARCH: Future work should continue to focus on longitudinal studies of fathering and the role of the NICU in encouraging involvement and parenting readiness.

Sagittal Plane Residual Deformity in Pediatric Type II Supracondylar Humerus Fractures.

INTRODUCTION: The incidence of residual deformity in the sagittal plane of the humerus (RDSPH) after nonoperative management of type II supracondylar humerus fractures (SCHFs), and the effects of such deformity on the overall arc of motion (AOM) of the elbow, are unknown. Our purpose was to analyze data collected prospectively on a large cohort of type II SCHF's to establish the incidence and extent of RDSPH, and the effects of the deformity on the elbow function, to further support our previously published recommendations on the treatment of type II SCHF. METHODS: The clinical data and radiographs of 1107 pediatric type II SCHFs enrolled in a prospective registry, and followed for a minimum of 8 weeks, were retrospectively reviewed. The radiographs obtained during the latest follow-up appointment were examined for the presence of RDSPH, as demonstrated by the anterior humeral line falling anterior or posterior relative to the center of the capitellum. The amount of RDSPH in the sagittal plane was then calculated. We compared the treatment outcome of elbows with and without RDSPH by assessing the patients' AOM, the arc of flexion (AOF), and relative arcs of motion (R-AOM) and relative arcs of flexion (R-AOF) (as compared with the unaffected, contralateral elbow). RESULTS: Overall, 799 (72.2%) fractures were treated nonsurgically, and 308 (27.8%) fractures were treated surgically. The overall incidence of RDSPH was 10.2%. None of the fractures managed operatively demonstrated residual deformity. The RDSPH was classified as mild in 35 fractures (3.2%), moderate in 64 fractures (5.7%) and severe in 14 fractures (1.3%). Therefore, the incidence of RDSPH in fractures treated nonoperatively was 14.1%. In fractures treated nonoperatively, the difference in AOM between those without (n=686) and with (n=113) RDSPH was <4 degrees (149.1 vs. 145.8 degrees, P=0.02). Those with and without RDSPH had a clinically similar AOF, with a mean difference of<4 degrees (134.5 vs. 137.9 degrees, P<0.0001). The differences in R-AOM and R-AOF between those with and without RDSPH were minimal (97.3% vs. 95.6% and 96.6% vs. 95.3%, respectively). A satisfactory outcome, defined as an R-AOM of at least 85% when compared with the unaffected, contralateral side at the latest follow-up, was achieved in 91% of fractures with RDSPH, and 93% of fractures without RDSPH. DISCUSSION AND CONCLUSION: The incidence of RDSPH in type II SCHF treated nonoperatively was 14%. In our cohort, nearly 99% of all RDSPH were mild to moderate. On the basis of the data presented in the current study, nonsurgical treatment of type II SCHF can provide a satisfactory recovery of AOM, AOF, R-AOM, and R-AOF, and a high rate of satisfactory outcomes, even in the presence of RDSPH.

The applicability of the UK Public Health Skills and Knowledge Framework to the practitioner workforce: lessons for competency framework development.

BACKGROUND: Many countries have developed competency frameworks for public health practice. While the number of competencies vary, frameworks cover similar knowledge and skills although they are not explicitly based on competency theory. METHODS: A total of 15 qualitative group interviews (of up to six people), were conducted with 51 public health practitioners in 8 local authorities to assess the extent to which practitioners utilize competencies defined within the UK Public Health Skills and Knowledge Framework (PHSKF). Framework analysis was applied to the transcribed interviews. RESULTS: The overall framework was seen positively although no participants had previously read or utilized the PHSKF. Most could provide evidence, although some PHSKF competencies required creative thinking to fit expectations of practitioners and to reflect variation across the domains of practice which are impacted by job role and level of seniority. Evidence from previous NHS jobs or education may be needed as some competencies were not regularly utilized within their current local authority role. CONCLUSIONS: Further development of the PHSKF is required to provide guidance on how it should be used for practitioners and other members of the public health workforce. Empirical research can help benchmark knowledge/skills for workforce levels so improving the utility of competency frameworks.

Responding to sustained poor outcomes in the management of non-communicable diseases (NCDs): an "incident control" approach is needed to improve and protect population health.

In 2017 Public Health England were asked to assist with investigating why 1-year cancer survival rates appeared lower than expected in a local area. We identified 50 premature deaths that surveillance data suggested we would not expect. These deaths highlighted a gap in recognising and responding to this kind of systematic non communicable disease (NCD) outcome variation. We hypothesise that the lack of a universally agreed systematic response to variations is not only counter-intuitive, but wholly unacceptable where non-communicable diseases (NCDs) rather than infectious diseases have become the leading causes of illness and death worldwide. In the United Kingdom (UK) alone over 89% of mortality in 2014 was attributable to NCDs. We argue that a new approach is urgently needed to turn the curve on NCD outcome variation to protect and improve the public's health. We set out a definition of an NCD "incident" and propose a phased approach that could be used to respond to local variation in NCD outcomes.Establishing parity of response for local variations in NCD outcomes and CD control is critically important. Although evidence shows that prevention and early intervention will make the biggest difference to NCD incidence, collective local whole health economy response, exploiting the wealth of surveillance data in real time, needs to be at the heart of responding to variations in NCD outcomes at a population level. We argue that local and national public health agencies should mandate a standardised 'incident' response to significant changes in outcomes from NCD to mitigate and reduce the loss of quality life.

Master or apprentice: rethinking entry points and training in Public Health.

Background: The Public Health workforce needs to adapt to a policy environment in which the need to improve public health is not only a key challenge but also has to be delivered within financial constraints. Methods: A total of 14 qualitative individual interviews or focus groups (of up to 10 people), were conducted with senior Public Health staff in 12 local authorities and in Public Health England in the Yorkshire and the Humber region of England. Thematic analysis was applied to the transcribed interviews. Results: Despite cuts in training budgets, junior staff were increasingly expected to deliver Public Health functions. There is also an absence of a career ladder for this core Public Health workforce. There were concerns that financial constraints would lead to reductions in Public Health skills with fewer staff to provide critical analysis. Formal qualifications were increasingly less valued. Instead staff were expected to have a broader skill set with an emphasis on experience. Entry points into Public Health careers need rethinking. There was an expectation that Public Health departments would 'grow their own'. Conclusions: Apprenticeship schemes could help maintain or enhance the professional status of Public Health and provide a training route for Public Health practitioners.

Changes in health care spending and quality 4 years into global payment.

BACKGROUND: Spending and quality under global budgets remain unknown beyond 2 years. We evaluated spending and quality measures during the first 4 years of the Blue Cross Blue Shield of Massachusetts Alternative Quality Contract (AQC). METHODS: We compared spending and quality among enrollees whose physician organizations entered the AQC from 2009 through 2012 with those among persons in control states. We studied spending changes according to year, category of service, site of care, experience managing risk contracts, and price versus utilization. We evaluated process and outcome quality. RESULTS: In the 2009 AQC cohort, medical spending on claims grew an average of $62.21 per enrollee per quarter less than it did in the control cohort over the 4-year period (P<0.001). This amount is equivalent to a 6.8% savings when calculated as a proportion of the average post-AQC spending level in the 2009 AQC cohort. Analogously, the 2010, 2011, and 2012 cohorts had average savings of 8.8% (P<0.001), 9.1% (P<0.001), and 5.8% (P=0.04), respectively, by the end of 2012. Claims savings were concentrated in the outpatient-facility setting and in procedures, imaging, and tests, explained by both reduced prices and reduced utilization. Claims savings were exceeded by incentive payments to providers during the period from 2009 through 2011 but exceeded incentive payments in 2012, generating net savings. Improvements in quality among AQC cohorts generally exceeded those seen elsewhere in New England and nationally. CONCLUSIONS: As compared with similar populations in other states, Massachusetts AQC enrollees had lower spending growth and generally greater quality improvements after 4 years. Although other factors in Massachusetts may have contributed, particularly in the later part of the study period, global budget contracts with quality incentives may encourage changes in practice patterns that help reduce spending and improve quality. (Funded by the Commonwealth Fund and others.).

Continuous-relief diffractive microlenses for laser beam focusing.

Microscale, continuous-profile, diffractive lenses have been fabricated and characterized. Lenses designed to operate at λ0 = 405 nm were created by focused ion beam milling of a glass substrate. The micro-structured profile was analysed by confocal microscopy and optical performance was quantified by measurements of the transmitted laser beam profile. Lenses of size 125 µm × 125 µm, containing up to 18 annuli and focusing at 400 µm, 450 µm and 500 µm have been made. Measured focused beams were in excellent agreement with the predicted performance. A maximum diffraction efficiency of 84 % and side-lobe suppression down to the 10-4 level can be achieved. The suitability of the lenses for interfacing with trappedion systems is outlined.

Health care spending and quality in year 1 of the alternative quality contract.

BACKGROUND: In 2009, Blue Cross Blue Shield of Massachusetts (BCBS) implemented a global payment system called the Alternative Quality Contract (AQC). Provider groups in the AQC system assume accountability for spending, similar to accountable care organizations that bear financial risk. Moreover, groups are eligible to receive bonuses for quality. METHODS: Seven provider organizations began 5-year contracts as part of the AQC system in 2009. We analyzed 2006-2009 claims for 380,142 enrollees whose primary care physicians (PCPs) were in the AQC system (intervention group) and for 1,351,446 enrollees whose PCPs were not in the system (control group). We used a propensity-weighted difference-in-differences approach, adjusting for age, sex, health status, and secular trends to isolate the treatment effect of the AQC in comparisons of spending and quality between the intervention group and the control group. RESULTS: Average spending increased for enrollees in both the intervention and control groups in 2009, but the increase was smaller for enrollees in the intervention group--$15.51 (1.9%) less per quarter (P=0.007). Savings derived largely from shifts in outpatient care toward facilities with lower fees; from lower expenditures for procedures, imaging, and testing; and from a reduction in spending for enrollees with the highest expected spending. The AQC system was associated with an improvement in performance on measures of the quality of the management of chronic conditions in adults (P<0.001) and of pediatric care (P=0.001), but not of adult preventive care. All AQC groups met 2009 budget targets and earned surpluses. Total BCBS payments to AQC groups, including bonuses for quality, are likely to have exceeded the estimated savings in year 1. CONCLUSIONS: The AQC system was associated with a modest slowing of spending growth and improved quality of care in 2009. Savings were achieved through changes in referral patterns rather than through changes in utilization. The long-term effect of the AQC system on spending growth depends on future budget targets and providers' ability to further improve efficiencies in practice. (Funded by the Commonwealth Fund and others.).

Training public health superheroes: five talents for public health leadership.

BACKGROUND: Public health leaders have been criticized for their policy stances, relationships with governments and failure to train the next generation. New approaches to the identification and training of public health leaders may be required. To inform these, lessons can be drawn from public health 'superheroes'; public health leaders perceived to be the most admired and effective by their peers. METHODS: Members and Fellows of the UK Faculty of Public Health were contacted via e-newsletter and magazine and asked to nominate their 'Public Health Superhero'. Twenty-six responses were received, nominating 40 different people. Twelve semi-structured interviews were conducted. Thematic analysis, based on 'grounded theory', was conducted. RESULTS: Five leadership 'talents' for public health were identified: mentoring-nurturing, shaping-organizing, networking-connecting, knowing-interpreting and advocating-impacting. CONCLUSIONS: Talent-based approaches have been effective for leadership development in other sectors. These talents are the first specific to the practice of public health and align with some aspects of existing frameworks. An increased focus on identifying and developing talents during public health training, as opposed to 'competency'-based approaches, may be effective in strengthening public health leadership. Further research to understand the combination and intensity of talents across a larger sample of public health leaders is required.