Discovery of lirafugratinib (RLY-4008), a highly selective irreversible small-molecule inhibitor of FGFR2.

Fibroblast growth factor receptor (FGFR) kinase inhibitors have been shown to be effective in the treatment of intrahepatic cholangiocarcinoma and other advanced solid tumors harboring FGFR2 alterations, but the toxicity of these drugs frequently leads to dose reduction or interruption of treatment such that maximum efficacy cannot be achieved. The most common adverse effects are hyperphosphatemia caused by FGFR1 inhibition and diarrhea due to FGFR4 inhibition, as current therapies are not selective among the FGFRs. Designing selective inhibitors has proved difficult with conventional approaches because the orthosteric sites of FGFR family members are observed to be highly similar in X-ray structures. In this study, aided by analysis of protein dynamics, we designed a selective, covalent FGFR2 inhibitor. In a key initial step, analysis of long-timescale molecular dynamics simulations of the FGFR1 and FGFR2 kinase domains allowed us to identify differential motion in their P-loops, which are located adjacent to the orthosteric site. Using this insight, we were able to design orthosteric binders that selectively and covalently engage the P-loop of FGFR2. Our drug discovery efforts culminated in the development of lirafugratinib (RLY-4008), a covalent inhibitor of FGFR2 that shows substantial selectivity over FGFR1 (~250-fold) and FGFR4 (~5,000-fold) in vitro, causes tumor regression in multiple FGFR2-altered human xenograft models, and was recently demonstrated to be efficacious in the clinic at doses that do not induce clinically significant hyperphosphatemia or diarrhea.

Discovery and Clinical Proof-of-Concept of RLY-2608, a First-in-Class Mutant-Selective Allosteric PI3Kα Inhibitor That Decouples Antitumor Activity from Hyperinsulinemia.

PIK3CA (PI3Kα) is a lipid kinase commonly mutated in cancer, including ∼40% of hormone receptor-positive breast cancer. The most frequently observed mutants occur in the kinase and helical domains. Orthosteric PI3Kα inhibitors suffer from poor selectivity leading to undesirable side effects, most prominently hyperglycemia due to inhibition of wild-type (WT) PI3Kα. Here, we used molecular dynamics simulations and cryo-electron microscopy to identify an allosteric network that provides an explanation for how mutations favor PI3Kα activation. A DNA-encoded library screen leveraging electron microscopy-optimized constructs, differential enrichment, and an orthosteric-blocking compound led to the identification of RLY-2608, a first-in-class allosteric mutant-selective inhibitor of PI3Kα. RLY-2608 inhibited tumor growth in PIK3CA-mutant xenograft models with minimal impact on insulin, a marker of dysregulated glucose homeostasis. RLY-2608 elicited objective tumor responses in two patients diagnosed with advanced hormone receptor-positive breast cancer with kinase or helical domain PIK3CA mutations, with no observed WT PI3Kα-related toxicities. SIGNIFICANCE: Treatments for PIK3CA-mutant cancers are limited by toxicities associated with the inhibition of WT PI3Kα. Molecular dynamics, cryo-electron microscopy, and DNA-encoded libraries were used to develop RLY-2608, a first-in-class inhibitor that demonstrates mutant selectivity in patients. This marks the advance of clinical mutant-selective inhibition that overcomes limitations of orthosteric PI3Kα inhibitors. See related commentary by Gong and Vanhaesebroeck, p. 204 . See related article by Varkaris et al., p. 227 . This article is featured in Selected Articles from This Issue, p. 201.

RLY-4008, the First Highly Selective FGFR2 Inhibitor with Activity across FGFR2 Alterations and Resistance Mutations.

Oncogenic activation of fibroblast growth factor receptor 2 (FGFR2) drives multiple cancers and represents a broad therapeutic opportunity, yet selective targeting of FGFR2 has not been achieved. Although the clinical efficacy of pan-FGFR inhibitors (pan-FGFRi) validates FGFR2 driver status in FGFR2 fusion-positive intrahepatic cholangiocarcinoma, their benefit is limited by incomplete target coverage due to FGFR1- and FGFR4-mediated toxicities (hyperphosphatemia and diarrhea, respectively) and the emergence of FGFR2 resistance mutations. RLY-4008 is a highly selective, irreversible FGFR2 inhibitor designed to overcome these limitations. In vitro, RLY-4008 demonstrates >250- and >5,000-fold selectivity over FGFR1 and FGFR4, respectively, and targets primary alterations and resistance mutations. In vivo, RLY-4008 induces regression in multiple xenograft models-including models with FGFR2 resistance mutations that drive clinical progression on current pan-FGFRi-while sparing FGFR1 and FGFR4. In early clinical testing, RLY-4008 induced responses without clinically significant off-isoform FGFR toxicities, confirming the broad therapeutic potential of selective FGFR2 targeting. SIGNIFICANCE: Patients with FGFR2-driven cancers derive limited benefit from pan-FGFRi due to multiple FGFR1-4-mediated toxicities and acquired FGFR2 resistance mutations. RLY-4008 is a highly selective FGFR2 inhibitor that targets primary alterations and resistance mutations and induces tumor regression while sparing other FGFRs, suggesting it may have broad therapeutic potential. See related commentary by Tripathi et al., p. 1964. This article is featured in Selected Articles from This Issue, p. 1949.

Non-traditional risk factors of progression of chronic kidney disease in adult population: a scoping review.

Chronic kidney disease (CKD) has become a public health concern over the last several years. Nowadays developed countries spend around 3% of their annual health-care budget on patients with CKD. According to the scientific community the most remarkable risk factors for CKD are diabetes and hypertension. Unknown CKD etiology has been reported as a global phenomenon including uncommon risk factors such as: dehydration, leptospirosis, heat stress, water quality, and others. This study aims to report non-traditional risk factors for ESRD based on a scoping review methodology. The scoping review methodology described by Arksey and O'Malley was used by performing an extensive review of the information. A total of 46 manuscripts were reviewed. The non-traditional ESRD risk factors are depicted based on six categories. Gender and ethnicity have been considered as risk factors for ESRD. Erythematous systemic lupus (ESL) is reported as an important risk factor for ESRD. Pesticide use has been an significant risk factor due to its effects on human and environmental health. Some compounds commonly used in homes against insects and plants are related to ESRD. Congenital and hereditary diseases in the urinary tract have been studied as a cause of ESRD in children and young adults. End-stage renal disease is a major concern for public health on a global level. As it can be seen, non-traditional risk factors are several and have different etiologies. It is necessary to put the issue on the table and add it to the public agenda in order to find multidisciplinary solutions.

Safety and efficacy of intra-arterial bone marrow mononuclear cell transplantation in patients with acute ischaemic stroke in Spain (IBIS trial): a phase 2, randomised, open-label, standard-of-care controlled, multicentre trial.

BACKGROUND: Pilot clinical trials have shown the safety of intra-arterial bone marrow mononuclear cells (BMMNCs) in stroke. However, the efficacy of different doses of intra-arterial BMMNCs in patients with acute stroke has not been tested in a randomised clinical trial. We aimed to show safety and efficacy of two different doses of autologous intra-arterial BMMNC transplantation in patients with acute stroke. METHODS: The IBIS trial was a multicentre phase 2, randomised, controlled, investigator-initiated, assessor-blinded, clinical trial, in four stroke centres in Spain. We included patients (aged 18-80 years) with a non-lacunar, middle cerebral artery ischaemic stroke within 1-7 days from stroke onset and with a National Institutes of Health Stroke Scale score of 6-20. We randomly assigned patients (2:1:1) with a computer-generated randomisation sequence to standard of care (control group) or intra-arterial injection of autologous BMMNCs at one of two different doses (2 × 106 BMMNCs/kg or 5 × 106 BMMNCs/kg). The primary efficacy outcome was the proportion of patients with modified Rankin Scale scores of 0-2 at 180 days in the intention-to-treat population, comparing each BMMNC dose group and the pooled BMMNC group versus the control group. The primary safety endpoint was the proportion of serious adverse events. This trial was registered at ClinicalTrials.gov, NCT02178657 and is completed. FINDINGS: Between April 1, 2015, and May 20, 2021, we assessed 114 patients for eligibility. We randomly assigned 77 (68%) patients: 38 (49%) to the control group, 20 (26%) to the low-dose BMMNC group, and 19 (25%) the high-dose BMMNC group. The mean age of participants was 62·4 years (SD 12·7), 46 (60%) were men, 31 (40%) were women, all were White, and 63 (82%) received thrombectomy. The median NIHSS score before randomisation was 12 (IQR 9-15), with intra-arterial BMMNC injection done a median of 6 days (4-7) after stroke onset. The primary efficacy outcome occurred in 14 (39%) patients in the control group versus ten (50%) in the low-dose group (adjusted odds ratio 2·08 [95% CI 0·55-7·85]; p=0·28), eight (44%) in the high-dose group (1·89 [0·52-6·96]; p=0·33), and 18 (47%) in the pooled BMMNC group (2·22 [0·72-6·85]; p=0·16). We found no differences in the proportion of patients who had adverse events or dose-related events, but two patients had a groin haematoma after cell injection in the low-dose BMMNC group. INTERPRETATION: Intra-arterial BMMNCs were safe in patients with acute ischaemic stroke, but we found no significant improvement at 180 days on the mRS. Further clinical trials are warranted to investigate whether improvements might be possible at different timepoints. FUNDING: Instituto de Salud Carlos III co-funded by the European Regional Development Fund/European Social Fund, Mutua Madrileña, and the Regional Ministry of Health of Andalusia.

Symptoms of tobacco dependence among middle and high school tobacco users - Data from the 2019-2020 National Youth Tobacco Survey.

INTRODUCTION: Previous studies have indicated that youth who use tobacco products, including cigarettes, cigars, and smokeless tobacco, demonstrate dependence symptoms. However, the tobacco marketplace has expanded dramatically in recent years, and few studies have examined dependence symptoms among youth who use novel products. This study combined 2019-2020 National Youth Tobacco Survey data to report the prevalence and determinants of tobacco dependence symptoms among U.S. middle and high school current (past 30-day) tobacco users. METHODS: Prevalence estimates were calculated to examine dependence outcomes and other covariates by user groups (single product users and multiple product users). Multivariable logistic regression analyses were used to identify independent predictors of tobacco dependence among current users of cigarettes, cigars (regular cigars, cigarillos, and little cigars), e-cigarettes, heated tobacco products, hookah, pipe tobacco, bidis, and smokeless tobacco products (chew, snuff, dip, snus, and dissolvables). RESULTS: Among current tobacco users, 15.7 % (95 % CI: 14.2-17.3) reported wanting to use tobacco within 30 min of waking and 28.3 % (95 % CI: 26.3-30.5) reported strong cravings for tobacco in the past 30 days. Nearly-two-thirds of current users were single product users, of which 80.5 % reported using e-cigarettes. Reporting of dependence symptoms was generally associated with multiple product use, higher frequency of use, earlier initiation age, and use of flavored products. CONCLUSIONS: Among U.S. adolescents, a considerable amount of current tobacco product users, even infrequent users, reported symptoms of dependence. These findings highlight the continued importance of prevention strategies for youth tobacco experimentation and progression to regular use.

Changes in Availability of ENDS: 2019-2020, U.S.

INTRODUCTION: Events during 2019 and 2020, such as the outbreak of e-cigarette, or vaping, product use‒associated lung injury; manufacturer product withdrawals; federal regulations; and coronavirus disease 2019, potentially affected the retail availability of ENDS in the U.S. Measuring changes in ENDS availability informs the understanding of the ENDS marketplace and contextualizes sales trends. METHODS: Joinpoint regression was used to estimate slope changes in the number of available ENDS in 2019 and 2020 and considered correspondence with tobacco marketplace events. Availability, the weekly number of unique universal product codes with nonzero sales, was derived from NielsenIQ scanner data. U.S. ENDS availability was modeled overall and by subproduct and flavor category within subproduct: mint, menthol, tobacco flavored, and undetermined. RESULTS: ENDS availability increased by 66% from January 2019 to December 2020. Availability decreased by 43% among prefilled cartridges and increased by 511% among disposables, both led by flavored varieties. During January 2020-February 2020, prefilled cartridge availability decreased by 23.71 universal product codes per week. During July 2020-August 2020, disposable availability increased by 27.90 universal product codes per week, led by flavored products. CONCLUSIONS: ENDS availability increased during 2019 through 2020, led by a rise in flavored disposables. Multiple slope changes in ENDS availability occurred, many coinciding with tobacco marketplace events. The slope of ENDS explicitly prioritized for federal enforcement (i.e., flavored prefilled cartridges) notably decreased in early 2020 and, soon thereafter, the slope of ENDS not explicitly prioritized for enforcement (e.g., flavored disposables) notably increased, suggesting an association with U.S. Food and Drug Administration's prioritized enforcement guidance.

Circulating microRNA after autologous bone marrow mononuclear cell (BM-MNC) injection in patients with ischemic stroke.

Previous studies have shown the potential of microRNAs (miRNA) in the pathological process of stroke and functional recovery. Bone marrow mononuclear cell (BM-MNC) transplantation improves recovery in experimental models of ischemic stroke that might be related with miRNA modifications. However, its effect on circulating miRNA has not been described in patients with stroke. We aimed to evaluate the circulating levels of miRNAs after autologous BM-MNC transplantation in patients with stroke. We investigate the pattern of miRNA-133b and miRNA-34a expression in patients with ischemic stroke included in a multicenter randomized controlled phase IIb trial (http://www.clinicaltrials.gov; unique identifier: NCT02178657). Patients were randomized to 2 different doses of autologous intra-arterial BM-MNC injection (2×106/kg or 5×106/kg) or control group within the first 7 days after stroke onset. We evaluate plasma concentration of miRNA-113b and miRNA-34a at inclusion and 4, 7, and 90 days after treatment. Thirteen cases (8 with 2×106/kg BM-MNC dose and 5 with 5×106/kg dose) and 11 controls (BM-MNC non-treated) were consecutively included. Mean age was 64.1±12.3 with a mean National Institutes of Health Stroke Scale score at inclusion of 14.5. Basal levels of miRNA were similar in both groups. miR-34a-5p and miR-133b showed different expression patterns. There was a significant dose-dependent increase of miRNA-34a levels 4 days after BM-MNC injection (fold change 3.7, p<0.001), whereas miRNA-133b showed a significant increase in the low-dose BM-MNC group at 90 days. Intra-arterial BM-MNC transplantation in patients with ischemic stroke seems to modulate early circulating miRNA-34a levels, which have been related to precursor cell migration in stroke and smaller infarct volumes.

Molecular Structures of Yeast Kinetochore Subcomplexes and Their Roles in Chromosome Segregation.

Kinetochore molecular architecture exemplifies "form follows function." The simplifications that generated the one-chromosome:one-microtubule linkage in point-centromere yeast have enabled strategies for systematic structural analysis and high-resolution visualization of many kinetochore components, leading to specific proposals for molecular mechanisms. We describe here some structural features that allow a kinetochore to remain attached to the end of a depolymerizing microtubule (MT) and some characteristics of the connections between substructures that permit very sensitive regulation by differential kinase activities. We emphasize in particular the importance of flexible connections between rod-like structural members and the integration of these members into a compliant cage-like assembly anchored on the MT by a sliding molecular ring.

Structure of the MIND Complex Defines a Regulatory Focus for Yeast Kinetochore Assembly.

Kinetochores connect centromeric nucleosomes with mitotic-spindle microtubules through conserved, cross-interacting protein subassemblies. In budding yeast, the heterotetrameric MIND complex (Mtw1, Nnf1, Nsl1, Dsn1), ortholog of the metazoan Mis12 complex, joins the centromere-proximal components, Mif2 and COMA, with the principal microtubule-binding component, the Ndc80 complex (Ndc80C). We report the crystal structure of Kluyveromyces lactis MIND and examine its partner interactions, to understand the connection from a centromeric nucleosome to a much larger microtubule. MIND resembles an elongated, asymmetric Y; two globular heads project from a coiled-coil shaft. An N-terminal extension of Dsn1 from one head regulates interactions of the other head, blocking binding of Mif2 and COMA. Dsn1 phosphorylation by Ipl1/Aurora B relieves this autoinhibition, enabling MIND to join an assembling kinetochore. A C-terminal extension of Dsn1 recruits Ndc80C to the opposite end of the shaft. The structure and properties of MIND show how it integrates phospho-regulatory inputs for kinetochore assembly and disassembly.

Conserved Tetramer Junction in the Kinetochore Ndc80 Complex.

The heterotetrameric Ndc80 complex establishes connectivity along the principal longitudinal axis of a kinetochore. Its two heterodimeric subcomplexes, each with a globular end and a coiled-coil shaft, connect end-to-end to create a ∼600 Å long rod spanning the gap from centromere-proximal structures to spindle microtubules. Neither subcomplex has a known function on its own, but the heterotetrameric organization and the characteristics of the junction are conserved from yeast to man. We have determined crystal structures of two shortened ("dwarf") Ndc80 complexes that contain the full tetramer junction and both globular ends. The junction connects two α-helical coiled coils through regions of four-chain and three-chain overlap. The complexity of its structure depends on interactions among conserved amino-acid residues, suggesting a binding site for additional cellular factor(s) not yet identified.

Long-Term Care Providers and services users in the United States: data from the National Study of Long-Term Care Providers, 2013-2014.

Long-term care services provided by paid, regulated providers are an important component of personal health care spending in the United States. This report presents the most current national descriptive results from the National Study of Long-Term Care Providers (NSLTCP), which is conducted by the Centers for Disease Control and Prevention's National Center for Health Statistics (NCHS). Data presented are drawn from multiple sources, primarily NCHS surveys of adult day services centers and residential care communities (covers 2014 data year); and administrative records obtained from the Centers for Medicare and Medicare Services (CMS) on home health agencies, hospices, and nursing homes (covers 2013 and 2014 data years). This report provides information on the supply, organizational characteristics, staffing, and services offered by paid, regulated providers of long-term care services; and the demographic, health, and functional composition of users of these services. Services users include residents of nursing homes and residential care communities, patients of home health agencies and hospices, and participants of adult day services centers. This report updates "Long-Term Care Services in the United States: 2013 Overview" (available from: http://www.cdc.gov/nchs/data/nsltcp/long_term_care_services_2013.pdf), which covered data years 2011 and 2012. In contrast, the title of this report and future reports will reflect the years of the data used rather than the publication year, in this case 2013 through 2014. A forthcoming companion product to this report, "Long-Term Care Providers and Services Users in the United States­State Estimates Supplement: National Study of Long-Term Care Providers, 2013­2014," contains tables and maps showing comparable state estimates for the national findings in this report, and will be available from: http://www.cdc.gov/nchs/ nsltcp/nsltcp_products.htm.

A Note on the Effect of Data Clustering on the Multiple-Imputation Variance Estimator: A Theoretical Addendum to , JOS.

Multiple imputation is a popular approach to handling missing data. Although it was originally motivated by survey nonresponse problems, it has been readily applied to other data settings. However, its general behavior still remains unclear when applied to survey data with complex sample designs, including clustering. Recently, Lewis et al. (2014) compared single- and multiple-imputation analyses for certain incomplete variables in the 2008 National Ambulatory Medicare Care Survey, which has a nationally representative, multistage, and clustered sampling design. Their study results suggested that the increase of the variance estimate due to multiple imputation compared with single imputation largely disappears for estimates with large design effects. We complement their empirical research by providing some theoretical reasoning. We consider data sampled from an equally weighted, single-stage cluster design and characterize the process using a balanced, one-way normal random-effects model. Assuming that the missingness is completely at random, we derive analytic expressions for the within- and between-multiple-imputation variance estimators for the mean estimator, and thus conveniently reveal the impact of design effects on these variance estimators. We propose approximations for the fraction of missing information in clustered samples, extending previous results for simple random samples. We discuss some generalizations of this research and its practical implications for data release by statistical agencies.

Differences in adult day services center participant characteristics by center ownership: United States, 2012.

KEY FINDINGS: Data from the National Study of Long-Term Care Providers. In 2012, more than one-third of participants in adult day services centers were younger than 65. A higher percentage of participants in nonprofit centers than in for-profit centers were younger than 65. About one-half of participants in adult day services centers were non-Hispanic white persons. A higher percentage of participants in for-profit centers than in nonprofit centers were Hispanic or were non-Hispanic and of a race other than black or white. Almost one-third of adult day services center participants had Alzheimer's disease or other dementias, and about one-quarter had a developmental disability. A lower percentage of participants in for-profit than in nonprofit centers had Alzheimer's disease or other dementias or a developmental disability. The 4,800 adult day services centers nationwide provide a variety of services to their 273,200 participants, the majority of whom are older adults and women (1). The number of for-profit adult day services centers has grown in recent years (2). The 1,900 for-profit centers, representing 40% of centers nationally, served nearly one-half (47%) of center participants in 2012 (3). Using data from the National Study of Long-Term Care Providers, this report presents selected characteristics of adult day services center participants in 2012 and compares the characteristics of participants in for-profit centers with those in nonprofit centers.

Differences in adult day services center characteristics by center ownership: United States, 2012.

KEY FINDINGS: Data from the National Study of Long-Term Care Providers. In 2012, 40% of the 4,800 adult day services centers were for-profit entities, serving nearly one-half of the 272,300 center participants. About 60% of adult day services centers used a standardized tool to screen for cognitive impairment, and about 20% used a standardized tool for depression screening. A greater percentage of for-profit than nonprofit centers used these tools. More than one-half of adult day services centers provided skilled nursing, therapeutic, and social work services, while less than one-half of centers provided mental health, pharmacy, and dental services. With the exception of social work services, a greater percentage of for-profit than nonprofit centers provided these services. Almost all adult day services centers provided daily transportation to and from the center. The most recent data estimate that 4,800 adult day services centers nationwide serve nearly a quarter million participants daily (1). Unlike other long-term care providers, such as nursing homes, home health agencies, hospices, and residential care communities, the majority of adult day services centers are nonprofit (1). However, for-profit ownership of adult day services centers appears to be increasing, from 27% in 2010 to 40% in 2012 (2). Using data from the National Study of Long-Term Care Providers, this report presents national estimates for characteristics of adult day services centers in 2012 and compares them by type of center ownership.

Implementación de la filosofía hooponopónica modificada en el programa terapéutico para pacientes que funcionan a nivel neurótico / Using the modification of the ho´oponopono philosophy in the therapeutic program for patients with neurotic levels

Se fundamenta la implementación de la filosofía hooponopónica, modificada en el programa terapéutico de los pacientes que funcionan a nivel neurótico en el Hospital de Día de Las Tunas, lo cual responde a una de las líneas de investigación del proyecto institucional FiloArtMed. La filosofía hooponopónica de fundamento idealista, exotérico y negador de la realidad, de orígenes milenarios en Hawái, fue modificada teniendo en cuenta el significado de sus palabras claves: perdóname, gracias, lo siento, te amo, de acuerdo a los preceptos dialécticos materialistas de la Medicina y la Psicología cubanas, insertándola en el programa terapéutico integral del hospital de día para pacientes neuróticos como técnica cognitiva conductual reguladora del comportamiento, rectorizando las terapias a manera de núcleo terapéutico. Se desecharon sus preceptos acientíficos tradicionales y se incluyó, además, el ejercicio de meditación, derivado de esta filosofía, basado en el principio de la neuroplasticidad cerebral, como técnica de relajación y de limpieza mental, fungiendo como la entrada B de la psicoterapia grupal sistémica de regulación emocional (AU)

This study deals with the use of the modification of the ho´oponopono philosophy in the therapeutic program for patients with neurotic levels who were treated at Las Tunas Psychiatric Day Hospital, as a response to the research lines of the FiloArtMed institutional project. The ho´oponopono philosophy of idealistic, exoteric and reality denier basis, which has millenary Hawaiian origins was modified taking into account the meaning of its keywords (forgive me, thanks, I am sorry and I love you) according to the dialectic materialistic precepts of Cuban Medicine and Psychology. Such modification is inserted in the therapeutic program of the day hospital for neurotic patients as a regulatory cognitive- behavioral therapy, ruling the therapies as a therapeutic nucleus. The traditional unscientific precepts were rejected and the meditation exercise derived from this philosophy and based on the brain neuroplasticity principle was included as a relaxation and mental cleaning technique, acting as the B entrance to systemic group psychotherapy of emotional regulation (AU)