Search

Purpose: This paper reports a review of couple-based behavioral HIV interventions conducted by the Social Intervention Group (SIG); and addresses gaps, future directions, and implications for couple-based HIV interventions. Method: We performed a literature review for SIG research on intervention and prevention studies involving couples/partners. Results: We identified nine couple-based interventions. Outcomes included reduced sexual and substance use-related risk behaviors and improved use of anti-retroviral treatment. We conducted these studies in diverse venues, including needle/syringe exchange programs, primary care clinics, and criminal justice settings. Conclusions: The findings of this review provide strong evidence for the efficacy of couple-based HIV interventions in reducing sexual HIV risks and linkage to HIV and substance-use treatment. SIG has advanced couple-based HIV intervention research science by improving study design, intervention core components, conceptual models, and implementation strategies; which have informed scientific directions and transformed couple-based HIV prevention research.

People in community corrections have rates of HIV and sexual risk behaviors that are much higher than the general population. Prior literature suggests that criminal justice involvement is associated with increased sexual risk behaviors, yet these studies focus on incarceration and use one-sided study designs that only collect data from one partner. To address gaps in the literature, this study used the Actor Partner-Interdependence Model with Structural Equation Modeling (SEM), to perform a dyadic analysis estimating individual (actor-only) partner-only, and dyadic patterns (actor-partner) of criminal justice involvement and greater sexual risks in a sample of 227 men on probation and their intimate partners in New York City, United States. Standard errors were bootstrapped with 10,000 replications to reduce bias in the significance tests. Goodness of fit indices suggested adequate or better model fit for all the models. Significant actor-only relationships included associations between exposures to arrest, misdemeanor convictions, time spent in jail or prison, felony convictions, lifetime number of incarceration events, prior conviction for disorderly conduct and increased sexual risk behaviors. Partner only effects included significant associations between male partners conviction for a violent crime and their female partners’ sexual risk behaviors. Men’s encounters with police and number of prior misdemeanors were associated with their own and intimate partners’ sexual risk behaviors. Women’s prior arrest was associated with their own and intimate partners’ sexual risk behaviors. The results from the present study suggest that men on probation and their intimate partners’ criminal justice involvement are associated with increased engagement in sexual risk behaviors. It is necessary to conduct greater research into developing dyadic sexual risk reduction and HIV/STI prevention interventions for people who are involved in the criminal justice system.

This paper describes the collaborative development of an agenda for research on e-assessment in university mathematics. We adapted an established approach to develop the agenda from the contributions of 22 mathematics education researchers, university teachers and learning technologists interested in this topic. The resulting set of 55 questions are grouped into 5 broad themes: errors and feedback, students, design and implementation, affordances, and mathematical skills. This agenda gives the framework for a programme of research aligned with practical concerns, that will contribute to both theoretical and practical development.

The following study investigates factors associated with discharge from OAT due to incarceration in a sample of 64,331 discharges in the United States. Multinomial regression investigated the association between demographic factors, prior arrest, referral source (i.e criminal justice agency) intravenous drug use, types of drug used, length of prior treatment and discharge due to incarceration compared to completing treatment or discharge due to other reasons. African Americans, Latinx, and Native Americans were at greater risk of discharge due to incarceration compared to whites. Referral to OAT from criminal justice agencies and self-referral was associated with increased risk of discharge from OAT due to incarceration compared to referral from a health care provider. Substance use of heroin, benzodiazepines, synthetic opioids, cocaine and non-prescription use of methadone were associated with discharge due to incarceration. Risk of discharge due to incarceration was higher for patients who reported intravenous drug use and who reported a co-morbid psychiatric problem. These findings enrich a nascent body of literature on mechanisms associated with attrition from OAT due to incarceration and emphasize the need for programs to divert people with OUD from incarceration to increase engagement and retention in OAT.

Community-driven responses are essential to ensure the adoption, reach and sustainability of evidence-based practices (EBPs) to prevent new cases of opioid use disorder (OUD) and reduce fatal and non-fatal overdoses. Most organizational approaches for selecting and implementing EBPs remain top-down and individually oriented without community engagement (CE). Moreover, few CE approaches have leveraged systems science to integrate community resources, values and priorities. This paper provides a novel CE paradigm that utilizes a data-driven and systems science approach; describes the composition, functions, and roles of researchers in CE; discusses unique ethical considerations that are particularly salient to CE research; and provides a description of how systems science and data-driven approaches to CE may be employed to select a range of EBPs that collectively address community needs. Finally, we conclude with scientific recommendations for the use of CE in research. Greater investment in CE research is needed to ensure contextual, equitable, and sustainable access to EBPs, such as medications for OUD (MOUD) in communities heavily impacted by the opioid epidemic. A data-driven approach to CE research guided by systems science has the potential to ensure adequate saturation and sustainability of EBPs that could significantly reduce opioid overdose and health inequities across the US.

It is widely recognised that coastal flood events can arise from combinations of extreme waves and sea levels. For flood risk analysis and the design of coastal structures it is therefore necessary to assess the joint probability of the occurrence of these variables. Traditional methods have involved the application of joint probability contours, defined in terms of extremes of sea conditions that can, if applied without correction factors, lead to the underestimation of flood risk and under-design of coastal structures. This paper describes the application of a robust multivariate statistical model to analyse extreme offshore waves, wind and sea levels around the coast of England. The approach described here is risk based in that it seeks to define extremes of response variables directly, rather than the joint extremes of sea conditions. The output of the statistical model comprises a Monte Carlo simulation of extreme events. These distributions of extreme events have been transformed from offshore to nearshore using a statistical emulator of a wave transformation model. The resulting nearshore extreme sea condition distributions have the potential to be applied for a range of purposes. The application is demonstrated using two structures located on the south coast of England.

Highlights • An approach to community implementation of evidence-based practices (EBPs) was needed. • The Opioid-overdose Reduction Continuum of Care Approach (ORCCA) was developed. • ORCCA outlines: evidence-based strategies, priority populations, community settings. • ORCCA is a flexible framework for EBP implementation to reduce opioid-overdose deaths., Background The number of opioid-involved overdose deaths in the United States remains a national crisis. The HEALing Communities Study (HCS) will test whether Communities That HEAL (CTH), a community-engaged intervention, can decrease opioid-involved deaths in intervention communities (n = 33), relative to wait-list communities (n = 34), from four states. The CTH intervention seeks to facilitate widespread implementation of three evidence-based practices (EBPs) with the potential to reduce opioid-involved overdose fatalities: overdose education and naloxone distribution (OEND), effective delivery of medication for opioid use disorder (MOUD), and safer opioid analgesic prescribing. A key challenge was delineating an EBP implementation approach useful for all HCS communities. Methods A workgroup composed of EBP experts from HCS research sites used literature reviews and expert consensus to: 1) compile strategies and associated resources for implementing EBPs primarily targeting individuals 18 and older; and 2) determine allowable community flexibility in EBP implementation. The workgroup developed the Opioid-overdose Reduction Continuum of Care Approach (ORCCA) to organize EBP strategies and resources to facilitate EBP implementation. Conclusions The ORCCA includes required and recommended EBP strategies, priority populations, and community settings. Each EBP has a “menu” of strategies from which communities can select and implement with a minimum of five strategies required: one for OEND, three for MOUD, and one for prescription opioid safety. Identification and engagement of high-risk populations in OEND and MOUD is an ORCCArequirement. To ensure CTH has community-wide impact, implementation of at least one EBP strategy is required in healthcare, behavioral health, and criminal justice settings, with communities identifying particular organizations to engage in HCS-facilitated EBP implementation.

Highlights • Community engagement can improve the uptake and sustainability of evidence-based practices (EBPs). • The Healing Communities Study seeks to show how community engagement can support the adoption of EBPs for addressing the opioid crisis. • To reduce opioid overdose deaths a community-engaged, data driven planning process is being implemented in 67 highly impacted communities., Background The implementation of evidence-based practices to reduce opioid overdose deaths within communities remains suboptimal. Community engagement can improve the uptake and sustainability of evidence-based practices. The HEALing Communities Study (HCS) aims to reduce opioid overdose deaths through the Communities That HEAL (CTH) intervention, a community-engaged, data-driven planning process that will be implemented in 67 communities across four states. Methods An iterative process was used in the development of the community engagement component of the CTH. The resulting community engagement process uses phased planning steeped in the principles of community based participatory research. Phases include: 0) Preparation, 1) Getting Started, 2) Getting Organized, 3) Community Profiles and Data Dashboards, 4) Community Action Planning, 5) Implementation and Monitoring, and 6) Sustainability Planning. Discussion The CTH protocol provides a common structure across the four states for the community-engaged intervention and allows for tailored approaches that meet the unique needs or sociocultural context of each community. Challenges inherent to community engagement work emerged early in the process are discussed. Conclusion HCS will show how community engagement can support the implementation of evidence-based practices for addressing the opioid crisis in highly impacted communities. Findings from this study have the potential to provide communities across the country with an evidence-based approach to address their local opioid crisis; advance community engaged research; and contribute to the implementation, sustainability, and adoption of evidence-based practices. Trial registration ClinicalTrials.gov (NCT04111939).

BackgroundThis study examines the relationship between experiencing intimate partner violence (IPV), exposure to prior childhood adversity, lifetime adverse experiences, drug-related relationship dependencies with intimate partners and overdose, hospitalization for drug use, friends and family members who overdosed and witnessing overdose.MethodologyThis paper included a sample of 201 women who use drugs in heterosexual relationships with criminal justice-involved men in New York City. We included measures of experiencing overdose, hospitalization for drug use, witnessing overdose, and having friends and family who overdosed. Intimate partner violence consisted of either 1) none/verbal only, 2) moderate and 3) severe abuse. Dichotomous indicators of drug-related relationship dependencies included financial support, drug procurement, splitting and pooling drugs. A scale measured cumulative exposure to childhood adversity and lifetime exposures to adverse events. This paper hypothesized that experiencing moderate and severe IPV, drug-related dependencies and exposure to prior childhood and lifetime adversity would be associated with a greater risk of experiencing overdose, hospitalization for drug use, witnessing overdose and having friends and family members who overdosed. Generalized linear modeling with robust variance estimated relative risk ratios that accounted for potential bias in confidence intervals and adjusted for race, ethnicity, education and marital status.ResultsWe found experiencing moderate or severe IPV was associated with ever being hospitalized for drug use and having a family member who experienced overdose. Experiencing moderate IPV was associated with increased risk of witnessing overdose, Partner drug dependencies were associated with overdose, ever being hospitalized for drug use, witnessing overdose, and having a family member or friend who experienced overdose. Childhood and lifetime adversity exposures were significantly associated with increased risk of overdose, ever being hospitalized for drug use, ever witnessing overdose and having a friend and family member who overdosed.ConclusionFindings underscore the intersection of experiencing IPV and drug-related relationship dependencies, childhood adversity and lifetime adversity in shaping experiences of and witnessing overdose among women who use drugs. They highlight the urgent need to address IPV, adversity experiences and drug-related relationship dependencies in overdose prevention for women who use drugs.

Background Multiple drug and sexual risk behaviors among people who inject drugs (PWID) in intimate relationships increase the risk of HIV and HCV transmission. Using data on PWID in intimate partnerships in Almaty, Kazakhstan, this study performed latent class analysis (LCA) on drug and sexual risk behaviors and estimated associations between dyadic relationship factors and membership in latent classes. Methods LCA was performed on a sample of 510 PWID (181-females/FWID, 321-males/MWID) to identify levels of drug and sexual risk behaviors. Generalized structural equation modeling with multinomial regressions estimated associations between relationship factors (length risk reduction communication, risk reduction self-efficacy) and class membership after adjusting for substance use severity, overdose, depression, binge drinking, intimate partner violence, structural factors, and sociodemographic characteristics. Models were sex-stratified to include FWID and PWID. Results A 3-class model best fit the data and consisted of low, medium, and high-risk classes. GSEM found that greater injection self-efficacy was associated with a lower likelihood of membership in the high-risk class for PWID and FWID. For MWID, greater length of the relationship was associated with a lower likelihood of membership in the medium-risk class. Greater relationship communication was associated with increased risk of membership in the high-risk latent class for MWID. Conclusions Future research must investigate if increasing risk reduction and safe sex self-efficacy could reduce drug and sexual risk behaviors and HIV transmission among PWID and their intimate partners. Interventions are needed that reduce power inequities within relationships as a method of increasing self-efficacy, particularly among women.

The design of breakwaters and other coastal structures around the UK requires the assessment of the joint probability of extreme waves and sea levels. There is a standard simplified approach applied within the UK that uses joint exceedance contours of waves and sea levels. This simplified approach can be non-conservative and lead to the under-design of coastal structures unless correction factors are applied. These limitations have been known for a number of years and alternative, more robust, methods have been applied in the past. These alternative methods are risk-based as they enable the probability of the consequence (structural or serviceability failure) to be established. The more robust methods are not, however, widely used in current practice. This paper describes the development of a statistically robust nearshore multivariate extremes data set around the coastline of England. The dataset can, in principle, be used to undertake risk-based design of structures, thereby overcoming the limitations of existing practice.

Background Kazakhstan and other countries in Central Asia are experiencing a rapidly growing HIV epidemic, which has historically been driven by injection drug use, but is more recently being fueled by heterosexual transmission. Methods This paper examines HIV and HCV infection, as well as sexual and drug-related risks among female partners of men who inject drugs (MWID), comparing females who inject drugs (FWID) to non-injecting female partners on socio-demographic, relationship context, and structural characteristics. Results The prevalence rate of HIV was 30.1% among FWID and 10.4% among non-IDU female partners of MWID. The prevalence rate of HCV was 89.8% among FWID and 14.8% among female non-IDUs. Less than one-fifth of all female participants had access to HIV education and services or harm reduction programs. Although high rates of non-injection drug use and sexual risk behaviors were found among both FWID and non-injecting female partners of MWID, we found that FWID were more likely to be HIV seropositive (aRR=3.03; 95% CI=1.78, 5.18) and HCV seropositive than non-IDU females (aRR=6.05; 95% CI=4.05, 9.04), were more likely to have used alcohol or drugs before sex (aRR=1.67; 95% CI=1.40, 2.00), and were more likely to have used sedatives, barbiturates, tranquilizers, sleeping pills, or painkillers that were not prescribed by a physician (aRR=17.45; 95% CI=8.01, 38.01). Conclusion Given the spread of the HIV epidemic to heterosexual partners in Kazakhstan, more attention is needed in research, prevention, and policies regarding female partners of male injection drug users.

Background: This study examines predictors of partner notification (PN) confirmed by a test counselor among people who inject drugs (PWID) and their sexual partners in Kazakhstan.Methods: We used baseline data from an HIV couple-based intervention study, restricting the sample to couples where both partners knew they were hepatitis C virus (HCV)-positive prior to participation in the study (N = 136 individuals). Cross-tabulation and logistic regression were used to examine predictors of PN, including socio-demographic characteristics, sexual and drug risk behaviors, and access to health services.Results: Of the sample, 68 (50%) participants notified their partners of their HCV status. PN was associated with participation in a needle/syringe exchange program and sexually transmitted infection counseling or education in the past 6 months. In the adjusted model, concurrent HIV infection (OR = 2.4, p

HCV infection is a serious concern among people who inject drugs. Despite imposing a major disease burden in countries with high rates of injection drug use such as Kazakhstan, other Central Asian and East Asian countries, Eastern Europe, and Russia, HCV remains an understudied issue. This study includes 728 individuals (364 couples) from Almaty, Kazakhstan, where at least one member of the dyad reported recent injection drug use. Participants were recruited to participate in a couple-based HIV prevention study. We examine the prevalence of HCV and co-infections between HCV and HIV, correlates of HCV, and the association between HCV prevalence and sexual behaviors after adjusting for non-injection illicit drug use, unsafe injection behaviors, and socio-demographic characteristics. Of the total sample, 90.2% of people who had ever injected drugs were diagnosed HCV positive and 15.5% of those who did not report injecting drugs were HCV positive. The prevalence of HIV and HCV co-infection among the total sample was 20.7%. In the multivariate analysis, having sex while high on alcohol and/or drugs was associated with HCV after adjusting for unsafe injection practices, non-injection drug use, and socio-demographic characteristics (PR=1.20; 95% CI=1.04, 1.39). The high rates of HCV and co-infection of HIV and HCV found in the study underscore the need for attention to HCV testing and treatment among people who inject drugs in Kazakhstan and their sex partners. Keywords: HCV, Injection drug use, Central Asia, HIV, Kazakhstan.

The impacts of storm surges represent an increasing risk to the world's coastlines. Coastal planners require accurate estimates of flood risk in order to provide suitable defensive measures. Therefore a reliable methodology is required for the estimation of extreme sea-level probabilities at high spatial resolution along coastlines. This paper describes a new method for estimating these probabilities, with application to the UK coastline. The method consists of two components: the estimation of extreme sea-levels by applying a newly developed statistical method, termed the Skew Surge Joint Probability Method, with tide gauge records, and the use of hindcast sea-levels to dynamically interpolate these estimates around complex coastlines. The skew surge parameter is a more reliable indicator of meteorological impacts on sea-level than the non-tidal residual used in the Revised Joint Probability Method, as previously used in the United Kingdom. The method has been applied to the UK coastline to provide a database of extreme sea-level probabilities for the Environment Agency for England and Wales and the Scottish Environment Protection Agency. The database will be used to inform coastal defense strategy, flood mapping and forecasting and to support policy, implementation and operational decision-making.

The process of mitosis involves a comprehensive reorganization of the cell: chromosomes condense, the nuclear envelope breaks down, the mitotic spindle is assembled, cells round up and release their ties to the substrate and so on and so forth. This reorganization is triggered by the activation of the protein kinase, Cyclin-Dependent Kinase 1 (CDK1). The end of mitosis is marked by the proteolysis of the cyclin subunit of CDK1, which terminates kinase activity. At this point, the phosphate moieties that altered the properties of hundreds of proteins to bring about the cellular reorganization are removed by protein phosphatases. At least one protein phosphatase, PP2A-B55, is completely shut off in mitosis. Depletion of this particular form of PP2A accelerates entry into mitosis, and blocks exit from mitosis. Control of this phosphatase is achieved by an inhibitor protein (α-endosulfine or ARPP-19) that becomes inhibitory when phosphorylated by a protein kinase called Greatwall, which is itself a substrate of CDK1. Failure to inhibit PP2A-B55 causes arrest of the cell cycle in G2 phase. I will discuss the role of this control mechanism in the control of mitosis.

Every year in the UK, many flood risk assessments are carried out on small catchments, typically draining areas of less than 25 km2 . Standard hydrological practice in all UK catchments is to apply the methods presented in the Flood Estimation Handbook (FEH) and its subsequent updates. FEH methods are practical, relatively easy to apply and based on extensive statistical analyses. However, uncertainties can be large, especially in atypical catchments, and small catchments can present unique challenges in terms of heavy urbanisation and rapid flood responses. Compared to larger catchments, small catchment flood data are limited. In this study, we use a dataset of annual maxima and digital catchment descriptors at 205 small catchments to benchmark the QMED and Q100 estimation performance of current UK flood estimation methods: the FEH statistical method, ReFH2 and MacDonald and Fraser’s method, in rural and urbanised catchments separately. All methods perform similarly in rural catchments overall, although MacDonald and Fraser’s method underestimates QMED in urbanised catchments. The methods show a larger factorial standard error against this small catchment dataset than they do against typical datasets of mixed-size catchments. Further work will evaluate the performance of ReFH2 in combination with the latest FEH13 rainfall model.

The nuclear-encoded Krebs cycle enzymes, fumarate hydratase (FH) and succinate dehydrogenase (SDHB, -C and -D), act as tumour suppressors. Germline mutations in FH predispose individuals to leiomyomas and renal cell cancer (HLRCC), whereas mutations in SDH cause paragangliomas and phaeochromocytomas (HPGL). In this study, we have shown that FH-deficient cells and tumours accumulate fumarate and, to a lesser extent, succinate. SDH-deficient tumours principally accumulate succinate. In situ analyses showed that these tumours also have over-expression of hypoxia-inducible factor 1alpha (HIF1alpha), activation of HIF1alphatargets (such as vascular endothelial growth factor) and high microvessel density. We found no evidence of increased reactive oxygen species in our cells. Our data provide in vivo evidence to support the hypothesis that increased succinate and/or fumarate causes stabilization of HIF1alpha a plausible mechanism, inhibition of HIF prolyl hydroxylases, has previously been suggested by in vitro studies. The basic mechanism of tumorigenesis in HPGL and HLRCC is likely to be pseudo-hypoxic drive, just as it is in von Hippel-Lindau syndrome.

‘Dividing cells pass through a regular sequence of cell growth and division, known as the cell cycle’, according to a college textbook of biology published in 1983 [[1][1]], 5 years before the underlying principles of control were first laid bare during 1988, the annus mirabilis of cell cycle

Activation of the cyclin-dependent kinase (Cdk1) cyclin B (CycB) complex (Cdk1:CycB) in mitosis brings about a remarkable extent of protein phosphorylation. Cdk1:CycB activation is switch-like, controlled by two auto-amplification loops—Cdk1:CycB activates its activating phosphatase, Cdc25, and inhibits its inhibiting kinase, Wee1. Recent experimental evidence suggests that parallel to Cdk1:CycB activation during mitosis, there is inhibition of its counteracting phosphatase activity. We argue that the downregulation of the phosphatase is not just a simple latch that suppresses futile cycles of phosphorylation/dephosphorylation during mitosis. Instead, we propose that phosphatase regulation creates coherent feed-forward loops and adds extra amplification loops to the Cdk1:CycB regulatory network, thus forming an integral part of the mitotic switch. These network motifs further strengthen the bistable characteristic of the mitotic switch, which is based on the antagonistic interaction of two groups of proteins: M-phase promoting factors (Cdk1:CycB, Cdc25, Greatwall and Endosulfine/Arpp19) and interphase promoting factors (Wee1, PP2A–B55 and a Greatwall counteracting phosphatase, probably PP1). The bistable character of the switch implies the existence of a CycB threshold for entry into mitosis. The end of G2 phase is determined by the point where CycB level crosses the CycB threshold for Cdk1 activation.

Cell cycle transitions depend on protein phosphorylation and dephosphorylation. Thediscovery of cyclin dependent kinases (CDKs) and their mode of activation by their cyclin partners explained many important aspects of cell cycle control. As the cell cycle is basically a series of recurrences of a defined set of events, protein phosphatases must obviously be as important as kinases. However, our knowledge about phosphatases lags well behind that of kinases. We still do not know exactly which phosphatase(s) is/are truly responsible for dephosphorylating CDK substrates, and we know very little about whether and how protein phosphatases are regulated. Here, we summarise our present understanding of the phosphatases that are important in the control of the cell cycle and pose the questions that need to be answered as regards the regulation of protein phosphatases.

Beyond the Greatwall Protein phosphorylation and dephosphorylation provide a central mechanism that controls the eukaryotic cell division cycle and entry of cells into mitosis. A form of protein phosphatase 2A (PP2A) has an important role inhibiting phosphorylation-dependent activation of cyclin-dependent kinase 1 (CDK1) itself and also dephosphorylating substrates of the active CDK1 that promote mitosis. PP2A activity is inhibited when another protein kinase, known as Greatwall, is activated (see the Perspective by Virshup and Kaldis ). Mochida et al. (p. 1670 ) and Gharbi-Ayachi et al. (p. 1673 ) searched for substrates of Greatwall that might participate in the cell cycle regulatory machinery. When phosphorylated by Greatwall, a pair of small related proteins, Arpp19 and α-endosulfine, inhibited activity of PP2A. These effects were critical for regulation of mitosis in Xenopus egg extracts and in human cancer cells. Greatwall itself is phosphorylated and activated by CDK1—thus, apparently contributing to a feed-forward loop that contributes to the switchlike commitment of cells to mitosis.

Introduction: Umbilical cord blood (UCB) grafts are the only option for a significant minority of patients who require hematopoietic stem cell transplantation (HSCT) but lack a suitable related or unrelated donor. While UCB can serve as a suitable 'off the shelf' graft for many patients, units with the best HLA match often contain low and sometimes insufficient numbers of CD34+ cells for use in transplantation, particularly for adult patients. Furthermore, UCB grafts contain lower CD34+ cells number compared to BM or PBSC grafts, which leads to longer engraftment times and a higher risk of graft failure. BM and PBSC grafts are usually injected intravenously, homing to the bone marrow after several hours in the circulation. During this time, CD34+ cells are lost in the lungs, liver and spleen with typically < 20% making it to the bone marrow.1 Investigators have sought to overcome the limitation of low cell dose in UCB grafts and loss of CD34+ cells in the circulation by injecting CD34+ cells directly into the bone marrow space. However, we have recently shown that conventional intrabone delivery methods used in investigational trials to transplant UCB, result in low-level retention of hematopoietic progenitor cells in the intrabone space. Recently, we developed an optimized intrabone (OIB) transplant method using computer controlled low pressure and low volume injection (controlled infusion rate Methods: Rhesus macaques received GCSF and plerixafor mobilization prior to apheresis. Products were CD34+ selected using Miltenyi beads. CD34+ cells were split equally for transduction with lentiviral vectors encoding the reporters GFP and YFP. After myeloablative conditioning with 10Gy total body irradiation, half the graft was injected directly intra-bone using the OIB method, with the other half of the autograft simultaneously being injected intravenously via slow iv push. Peripheral blood samples were measured daily by flow cytometry to assess the proportion of engrafting cells deriving from each source. To address whether OIB transplantation could allow engraftment of low CD34+ cell numbers, as found in UCB units, the cell doses transplanted in one rhesus macaque recipient were reduced to only 0.5 x 106CD34+ cells/kg. Results: CD34+ cells injected intrabone utilizing the OIB method engrafted in all 3 animals. However, flow cytometric analysis gating on GFP vs YFP positive neutrophils showed CD34+ cells injected utilizing OIB delivery did not engraft quicker than IV transplanted cells. Sequential monitoring of neutrophils over 30 days showed the contribution to hematopoiesis of OIB delivered cells was significantly lower than the cells injected IV (Table 1.) Conclusions: We developed a novel intrabone delivery system that optimizes the retention of CD34+ cells into the bone marrow space. Although autologous CD34+ cells injected using this OIB transplant method were capable of engrafting in rhesus macaques that had undergone myeloablative conditioning, they did not engraft faster and contributed less to hematopoiesis than CD34+ cells simultaneously transplanted using conventional IV infusion. These data raise questions over whether intrabone delivery, even when using techniques to optimize intrabone retention, has utility in improving CD34+ cell engraftment. References: 1. Van der Loo JC et al. Marrow and spleen seeding efficiencies of all murine hematopoietic stem cell subsets are de- creased by preincubation with hematopoietic growth factors. Blood. 1995; 85:2598-2606 2. Pantin et. al. Optimization of intrabone delivery of hematopoietic progenitor cells in a swine model using cell radiolabeling with 89zirconium. American J Transplant. 2015 Mar;15(3):606-17. Disclosures Davidson: Macrogenics: Employment. Pantin:NIH: Patents & Royalties: a patent application for an intrabone delivery device. Dunbar:National Institute of Health: Research Funding.

The Tipin/Tim1 complex plays an important role in the S-phase checkpoint and replication fork stability. However, the biochemical function of this complex is poorly understood. Using Xenopus laevis egg extract we show that Tipin is required for DNA replication in the presence of limiting amount of replication origins. Under these conditions the DNA replication defect correlates with decreased levels of DNA Pol alpha on chromatin. We identified And1, a Pol alpha chromatin-loading factor, as new Tipin-binding partner. We found that both Tipin and And1 promote stable binding of Pol alpha to chromatin and that this is required for DNA replication under unchallenged conditions. Strikingly, extracts lacking Tipin and And1 also show reduced sister chromatids cohesion. These data indicate that Tipin/Tim1/And1 form a complex that links stabilization of replication fork and establishment of sister chromatid cohesion. The EMBO Journal (2009) 28, 3681-3692. doi: 10.1038/emboj.2009.304; Published online 5 November 2009

Cyclin-dependent kinase 4 (CDK4)/cyclin D complexes are expressed early in the G 1 phase of the cell cycle and stimulate the expression of genes required for G 1 progression by phosphorylation of the product of the retinoblastoma gene, pRb. To elaborate the molecular pathway of CDK4 activation and substrate selection we have determined the structure of nonphosphorylated CDK4/cyclin D3. This structure of an authentic CDK/cyclin complex shows that cyclin binding may not be sufficient to drive the CDK active site toward an active conformation. Phosphorylated CDK4/cyclin D3 is active as a pRb kinase and is susceptible to inhibition by p27 Kip1 . Unlike CDK2/cyclin A, CDK4/cyclin D3 can be inactivated by treatment with λ-phosphatase, implying that phosphorylated T172 is accessible to a generic phosphatase while bound to a cyclin. Taken together, these results suggest that the structural mechanism of CDK4/cyclin D3 activation differs markedly from that of previously studied CDK/cyclin complexes.

Cell-cycle progression is regulated by cyclin-dependent kinases (CDKs). CDK1 and CDK2 can be also activated by noncyclin proteins named RINGO/Speedy, which were identified as inducers of the G2/M transition in Xenopus oocytes. However, it is unclear how XRINGO triggers M phase entry in oocytes. We show here that XRINGO-activated CDKs can phosphorylate specific residues in the regulatory domain of Myt1, a Wee1 family kinase that plays a key role in the G2 arrest of oocytes. We have identified three Ser that are major phosphoacceptor sites for CDK/XRINGO but are poorly phosphorylated by CDK/cyclin. Phosphorylation of these Ser inhibits Myt1 activity, whereas their mutation makes Myt1 resistant to inhibition by CDK/XRINGO. Our results demonstrate that XRINGO-activated CDKs have different substrate specificity than the CDK/cyclin complexes. We also describe a mechanism of Myt1 regulation based on site-specific phosphorylation, which is likely to mediate the induction of G2/M transition in oocytes by XRINGO.

Cell-cycle transitions in higher eukaryotes are regulated by different cyclin-dependent kinases (CDKs) and their activating cyclin subunits. Based on pioneering findings that a dominant-negative mutation of CDK1 blocks the cell cycle at G2-M phase, whereas dominant-negative CDK2 inhibits the transition into S phase, a model of cell-cycle control has emerged in which each transition is regulated by a specific subset of CDKs and cyclins. Recent work with gene-targeted mice has led to a revision of this model. We discuss cell-cycle control in light of overlapping and essential functions of the different CDKs and cyclins.

Objective: Various studies have shown that bone marrow stem cells can rescue mice from acute renal tubular damage under a conditioning advantage (irradiation or cisplatin treatment) favouring donor cell engraftment and regeneration; however, it is not known whether bone marrow cells (BMCs) can contribute to repair of acute tubular damage in the absence of a selection pressure for the donor cells. The aim of this study was to examine this possibility. Materials and methods: Ten‐week‐old female mice were assigned into control non‐irradiated animals having only vehicle treatment, HgCl(2)‐treated non‐irradiated mice, HgCl(2)‐treated non‐irradiated mice infused with male BMCs 1 day after HgCl(2), and vehicle‐treated mice with male BMCs. Tritiated thymidine was given 1 h before animal killing. Results: Donor BMCs could not alleviate non‐irradiated mice from acute tubular damage caused by HgCl(2), deduced by no reduction in serum urea nitrogen combined with negligible cell engraftment. However, donor BMCs could home to the bone marrow and spleen and display proliferative activity. This is the first report to show that despite no preparative myeloablation of recipients, engrafted donor BMCs can synthesize DNA in the bone marrow and spleen. Conclusions: Exogenous BMCs do not rescue non‐irradiated mice from acute renal tubular damage caused by HgCl(2), despite establishment of chimerism and cell proliferation in bone marrow and spleen.

Objective: Our previous studies have demonstrated that endogenous bone marrow cells (BMCs) contribute to renal tubular regeneration after acute tubular injury. The aim of this study was to examine which fraction of BMCs, haematopoietic lineage marrow cells (HLMCs) or mesenchymal stem cells (MSCs), are effective. Materials and methods: Six‐week‐old female mice were lethally irradiated and were transplanted with female enhanced green fluorescent protein‐positive (GFP(+)), plastic non‐adherent marrow cells (as a source of HLMCs) plus cloned cultured male GFP(−) MSCs. Four weeks later, they were assigned into two groups: control mice with vehicle treatment and mice treated with HgCl(2). Tritiated thymidine was given 1 h before animal killing which occurred at intervals over 2 weeks. Kidney sections were stained for a tubular epithelial marker, cell origin indicated by GFP immunohistochemistry or Y chromosome in situ hybridization; periodic acid‐Schiff staining was performed, and samples were subjected to autoradiography. One thousand consecutive renal tubular epithelial cells per mouse, in S phase, were scored as either female (indigenous) GFP(+) (HLMC‐derived) or male (MSC‐derived). Results: Haematopoietic lineage marrow cells and MSCs stably engrafted into bone marrow and spleen, but only HLMC‐derived cells, not MSCs, were found in the renal tubules and were able to undergo DNA synthesis after acute renal injury. A few MSCs were detected in the renal interstitium, but their importance needs to be further explored. Conclusion: Haematopoietic lineage marrow cells, but not cloned cultured MSCs, can play a role not only in normal wear‐and‐tear turnover of renal tubular cells, but also in repair after tubular injury.

Although the incidence of HIV among women on probation, parole and alternatives to incarceration programs is significant to public health, drivers of this concentrated epidemic among women under community corrections remain understudied. This study examined prevalence of HIV and sexually transmitted infections and the associations between substance use, socio-demographic factors and the prevalence of biologically-confirmed HIV and other sexually transmitted infections among a sample of 337 substance-using women recruited from community correction sites in New York City. Prevalence of HIV was 13% and sexually transmitted infections was 26% ( Chlamydia, trachomatis and Neisseria gonorrhea). After adjusting for covariates, HIV-positive women were 1.42 times more likely to use crack/cocaine than HIV-negative women (95% CI = 1.05–1.92). HIV-positive women were 25% less likely than HIV-negative women to report any unprotected vaginal and anal sex with their main partner (95% CI = 0.57–0.99). They were 70% less likely than HIV-negative women to report unprotected vaginal sex with a non-paying casual partner (95% CI = 0.1–0.9) and 22% less likely to report unprotected vaginal sex across all partners (95% CI = 0.61–0.99). Community corrections settings may be optimal venues to launch HIV/sexually transmitted infections prevention that have potential to reach and engage an ever-growing number of substance-using women.

Determining the sex of free-ranging cetaceans can be challenging. Sexual dimorphism among external features may allow inferences on sex, but such patterns may be difficult to detect and are often confounded by age and geographic variation. Dorsal fin images of 107 female and 54 male Australian humpback dolphins, Sousa sahulensis, from Western Australia (WA) and Queensland (QLD) were used to investigate sex, age and geographic differences in colouration, height/length quotient and number of notches. Adult males exhibited more dorsal fin notches (p0.001) and a significantly greater loss of pigmentation on the upper half of their dorsal fins (p0.001) than did adult females. These differences likely reflect that males experience a higher frequency and/or intensity of intraspecific aggression than females. In QLD, heavily spotted dorsal fins were more frequent among females than males (p0.001). Logistic regression analyses revealed that dorsal fin spotting and loss of pigmentation on the upper half of the dorsal fin provided the best model parameters for predicting the sex of sampled adults, with 97% accuracy. This technique offers a rapid, non-invasive method for predicting sex in Australian humpback dolphins, which could potentially be applied to populations throughout their range. In contrast to adults, presumed immature animals showed little or no loss of pigmentation or spotting; however, the rate of development of these features remains unknown. There were pronounced differences between QLD and WA in the intensity of spotting on dorsal fins and the extent of pigmentation loss around the posterior insertion and trailing edge of the dorsal fin. While based on a limited sample size, these geographic differences may have conservation implications in terms of population subdivision and should be investigated further.