Application of Laser-Induced Acoustic Desorption for Molecular Rotational Resonance Spectroscopy.

Molecular Rotational Resonance (MRR) spectroscopy is a uniquely precise tool for the determination of molecular structures of volatile compounds in mixtures, as the characteristic rotational transition frequencies of a molecule are extremely sensitive to its 3D structure through the moments of inertia in a three-dimensional coordinate system. This enables identification of the compounds based on just a few parameters that can be calculated, as opposed to, for example, mass spectrometric data, which often require expert analysis of 10-20 different signals and the use of many standards/model compounds. This paper introduces a new sampling technique for MRR, laser-induced acoustic desorption (LIAD), to allow the vaporization of nonvolatile and thermally labile analytes without the need for excessive heating or derivatization. In this proof-of-concept study, LIAD was successfully coupled to an MRR instrument to conduct measurements on seven compounds with differing polarities, molecular weights, and melting and boiling points. Identification of three isomers in a mixture was also successfully performed using LIAD/MRR. Based on these results, LIAD/MRR is demonstrated to provide a powerful approach for the identification of nonvolatile and/or thermally labile analytes with molecular weights up to 600 Da in simple mixtures, which does not require the use of reference compounds. In the future, applications to more complex mixtures, such as those relevant to pharmaceutical research, and quantitative aspects of LIAD/MRR will be reported.

Development of ISB 1442, a CD38 and CD47 bispecific biparatopic antibody innate cell modulator for the treatment of multiple myeloma.

Antibody engineering can tailor the design and activities of therapeutic antibodies for better efficiency or other advantageous clinical properties. Here we report the development of ISB 1442, a fully human bispecific antibody designed to re-establish synthetic immunity in CD38+ hematological malignancies. ISB 1442 consists of two anti-CD38 arms targeting two distinct epitopes that preferentially drive binding to tumor cells and enable avidity-induced blocking of proximal CD47 receptors on the same cell while preventing on-target off-tumor binding on healthy cells. The Fc portion of ISB 1442 is engineered to enhance complement dependent cytotoxicity, antibody dependent cell cytotoxicity and antibody dependent cell phagocytosis. ISB 1442 thus represents a CD47-BsAb combining biparatopic targeting of a tumor associated antigen with engineered enhancement of antibody effector function to overcome potential resistance mechanisms that hamper treatment of myeloma with monospecific anti-CD38 antibodies. ISB 1442 is currently in a Phase I clinical trial in relapsed refractory multiple myeloma.

The Methyltransferases METTL7A and METTL7B Confer Resistance to Thiol-Based Histone Deacetylase Inhibitors.

Histone deacetylase inhibitors (HDACi) are part of a growing class of epigenetic therapies used for the treatment of cancer. Although HDACis are effective in the treatment of T-cell lymphomas, treatment of solid tumors with this class of drugs has not been successful. Overexpression of the multidrug resistance protein P-glycoprotein (P-gp), encoded by ABCB1, is known to confer resistance to the HDACi romidepsin in vitro, yet increased ABCB1 expression has not been associated with resistance in patients, suggesting that other mechanisms of resistance arise in the clinic. To identify alternative mechanisms of resistance to romidepsin, we selected MCF-7 breast cancer cells with romidepsin in the presence of the P-gp inhibitor verapamil to reduce the likelihood of P-gp-mediated resistance. The resulting cell line, MCF-7 DpVp300, does not express P-gp and was found to be selectively resistant to romidepsin but not to other HDACis such as belinostat, panobinostat, or vorinostat. RNA-sequencing analysis revealed upregulation of the mRNA coding for the putative methyltransferase, METTL7A, whose paralog, METTL7B, was previously shown to methylate thiol groups on hydrogen sulfide and captopril. As romidepsin has a thiol as the zinc-binding moiety, we hypothesized that METTL7A could inactivate romidepsin and other thiol-based HDACis via methylation of the thiol group. We demonstrate that expression of METTL7A or METTL7B confers resistance to thiol-based HDACis and that both enzymes are capable of methylating thiol-containing HDACis. We thus propose that METTL7A and METTL7B confer resistance to thiol-based HDACis by methylating and inactivating the zinc-binding thiol.

Acute influence of resistance exercise on basketball shooting mechanics and accuracy.

The purpose of the present study was to examine the acute impact of resistance exercise on basketball shooting mechanics and accuracy. Ten resistance-trained recreationally active men with previous basketball playing experience (x̄ ± SD; height = 182.6 ± 9.7 cm; body mass = 79.2 ± 13.9 kg; age = 25.6 ± 5.5 years) performed control, upper-body, and lower-body training sessions in randomized order followed by 5 sets of stationary free-throw (4.57 m), two-point (5.18 m) and three-point (6.75 m) basketball shooting drills in 30 min time increments. Each testing session was separated 3-7 days apart. Kinematic variables during both the preparatory and release phases of the shooting motion were derived from a high-definition camera recording at 120 fps positioned 10 m away perpendicular to the participant's shooting plane of motion. Restricted maximum likelihood linear mixed-effects model analysis revealed that a combination of all fixed effects could account for <1% of the total variance in each dependent variable pertaining to basketball shooting mechanics. A 9.9-11.8% decrease in two-point and three-point shooting accuracy was observed immediately following an upper-body training session. However, the observed performance suppression disappeared 30 min post-exercise completion. Overall, the findings suggest that performing upper-body or lower-body resistance training prior to on-court practice sessions has no impact on free-throw, two-point, and three-point biomechanical parameters examined in the present study and a minor acute impact on mid-range and long-range shooting accuracy in male basketball players.

High-throughput cytochrome P450 loss and metabolic intermediate complex assays to aid in designing out of CYP3A inactivation.

Time-dependent inactivation (TDI) of cytochrome P450 (CYP) enzymes may result in clinical drug-drug interactions (DDIs). Therefore, designing out of CYP TDI prior to advancing a compound to clinical development is highly desirable. As TDI of CYP3A is a common occurrence in small molecule drug discovery, high-throughput methods are sought to help identify the mechanism of inactivation and enable design strategies to mitigate CYP3A TDI. CYP inactivation via modification or destruction of the prosthetic heme group results in loss of the ability of the enzyme to bind carbon monoxide. Additionally, formation of a tight binding complex with the heme iron, referred to as a metabolic intermediate (MI) complex, also results in enzyme inactivation. The methods described herein provide a high-throughput means of identifying and comparing compounds for their ability to inactivate via destruction/modification of the heme via loss of the ability to bind carbon monooxide, as well as via formation of an MI complex.

Salivary Testosterone and Cortisol Changes During a Game in Professional Male Basketball Players.

ABSTRACT: Cabarkapa, D, Eserhaut, DA, Cabarkapa, DV, Philipp, NM, and Fry, AC. Salivary testosterone and cortisol changes during a game in professional male basketball players. J Strength Cond Res 37(8): 1687-1691, 2023-The purpose of this study was to examine acute changes in salivary testosterone (T), cortisol (C), and testosterone-to-cortisol ratio (T/C) during a simulated 5-on-5 basketball game. Seven professional male basketball players volunteered to participate in this study. Repeated-measures analysis design was used to examine changes in hormonal concentrations across 8 testing time points: immediately upon arrival to the gymnasium-baseline (BS); post-warm-up (PW); post-first (P1Q), second (P2Q), third (P3Q), and fourth quarter (P4Q); and 30 (P30) and 60 minutes (P60) postgame. The findings of this study indicate that a simulated 5-on-5 basketball game provoked significant changes in salivary T, C, and T/C. When compared to the BS levels (x̄ ± SD [nmol·L-1]; 6.72 ± 2.53), salivary C concentration experienced a notable increase P3Q (16.20 ± 7.70) and remained elevated throughout the rest of the sampling periods, with values failing to return to BS levels P60 (11.88 ± 5.58). Conversely, a significant increase in salivary T occurred P1Q (0.76 ± 0.21) when compared to the BS levels (0.58 ± 0.12) and remained elevated up to P30 (0.75 ± 0.20), with values returning to BS levels P60 (0.63 ± 0.14). In addition, despite no significant intragame alterations, T/C exhibited a notable decrease P30 (0.06 ± 0.02) and P60 (0.07 ± 0.04), when compared to BS values (0.10 ± 0.04). Overall, these findings provide additional insight into the physiological stress that basketball players are exposed to during 5-on-5 competitive play and can be used to appropriately adjust and monitor training loads to optimize recovery and on-court basketball performance.

Repeat sprint fatigue and altered neuromuscular performance in recreationally trained basketball players.

The primary aim of the present study was to investigate how the fatigue induced through a repeat sprint protocol acutely affected different measures of neuromuscular performance. Recreationally trained basketball players (n = 25) volunteered to participate in the study, and performed three countermovement jumps (CMJ), as well as three drop jumps (DJ) prior to a fatiguing repeat sprint protocol. These procedures were repeated two minutes, and 15 minutes, following the protocol. Various force-time metrics were extracted from the jump tasks, and linear mixed models with subject ID as the random factor, and time as the fixed factor were used to investigate changes across the three time points. To account for the performance during the repeat sprint protocol, a second, two factor model was performed with time and repeat sprint ability (RSA) as the fixed factors. Study results indicated that the sample as a whole merely experienced fatigue-induced decreases in jump height from pre-repeat sprint ability protocol (pre-RSA) within the CMJ compared to two minutes post-repeat sprint ability protocol (post-RSA1) and 15 minutes post-repeat sprint ability protocol (post-RSA2), while jump height within the DJ was only significantly different from pre-RSA at post-RSA1. Further, despite the implementation of the fatiguing RSA protocol, over the course of the three time-points, participants seemed to perform the two jump tasks more efficiently, seen through significantly lower contraction times, greater eccentric (ECC) peak power, and greater ECC mean deceleration force within the CMJ following the RSA task. The two-factor model revealed that several significant time*RSA interactions were found for metrics such as ECC peak velocity and peak power in the CMJ, as well as reactive strength index in the DJ. This suggests that the level of RSA influenced changes across CMJ and DJ characteristics and should be accounted for when interpreting fatigue-induced changes in neuromuscular performance.

METTL7A (TMT1A) and METTL7B (TMT1B) Are Responsible for Alkyl S-Thiol Methyl Transferase Activity in Liver.

S-methylation of drugs containing thiol-moieties often alters their activity and results in detoxification. Historically, scientists attributed methylation of exogenous aliphatic and phenolic thiols to a putative S-adenosyl-L-methionine (SAM)-dependent membrane-associated enzyme referred to as thiol methyltransferase (TMT). This putative TMT appeared to have a broad substrate specificity and methylated the thiol metabolite of spironolactone, mertansine, ziprasidone, captopril, and the active metabolites of the thienopyridine prodrugs, clopidogrel, and prasugrel. Despite TMT's role in the S-methylation of clinically relevant drugs, the enzyme(s) responsible for this activity remained unknown. We recently identified methyltransferase-like protein 7B (METTL7B) as an alkyl thiol methyltransferase. METTL7B is an endoplasmic reticulum-associated protein with similar biochemical properties and substrate specificity to the putative TMT. Yet, the historic TMT inhibitor 2,3-dichloro-α-methylbenzylamine (DCMB) did not inhibit METTL7B, indicating that multiple enzymes contribute to TMT activity. Here we report that methyltransferase-like protein 7A (METTL7A), an uncharacterized member of the METTL7 family, is also a SAM-dependent thiol methyltransferase. METTL7A exhibits similar biochemical properties to METTL7B and putative TMT, including inhibition by DCMB (IC50 = 1.17 µM). Applying quantitative proteomics to human liver microsomes and gene modulation experiments in HepG2 and HeLa cells, we determined that TMT activity correlates closely with METTL7A and METTL7B protein levels. Furthermore, purification of a novel His-GST-tagged recombinant protein and subsequent activity experiments prove that METTL7A can selectively methylate exogenous thiol-containing substrates, including 7α-thiospironolactone, dithiothreitol, 4-chlorothiophenol, and mertansine. We conclude that the METTL7 family encodes for two enzymes, METTL7A and METTL7B, which are now renamed thiol methyltransferase 1A (TMT1A) and thiol methyltransferase 1B (TMT1B), respectively, that are responsible for thiol methylation activity in human liver microsomes. SIGNIFICANCE STATEMENT: We identified methyltransferase-like protein 7A (thiol methyltransferase 1A) and methyltransferase-like protein 7B (thiol methyltransferase 1B) as the enzymes responsible for the microsomal alkyl thiol methyltransferase (TMT) activity. These are the first two enzymes directly associated with microsomal TMT activity. S-methylation of commonly prescribed thiol-containing drugs alters their pharmacological activity and/or toxicity, and identifying the enzymes responsible for this activity will improve our understanding of the drug metabolism and pharmacokinetic (DMPK) properties of alkyl- or phenolic thiol-containing therapeutics.

Inter-Device Reliability of a Three-Dimensional Markerless Motion Capture System Quantifying Elementary Movement Patterns in Humans.

With advancements in technology able to quantify wide-ranging features of human movement, the aim of the present study was to investigate the inter-device technological reliability of a three-dimensional markerless motion capture system (3D-MCS), quantifying different movement tasks. A total of 20 healthy individuals performed a test battery consisting of 29 different movements, from which 214 different metrics were derived. Two 3D-MCS located in close proximity were utilized to quantify movement characteristics. Independent sample t-tests with selected reliability statistics (i.e., intraclass correlation coefficient (ICC), effect sizes, and mean absolute differences) were used to evaluate the agreement between the two systems. The study results suggested that 95.7% of all metrics analyzed revealed negligible or small between-device effect sizes. Further, 91.6% of all metrics analyzed showed moderate or better agreement when looking at the ICC values, while 32.2% of all metrics showed excellent agreement. For metrics measuring joint angles (198 metrics), the mean difference between systems was 2.9 degrees, while for metrics investigating distance measures (16 metrics; e.g., center of mass depth), the mean difference between systems was 0.62 cm. Caution is advised when trying to generalize the study findings beyond the specific technology and software used in this investigation. Given the technological reliability reported in this study, as well as the logistical and time-related limitations associated with marker-based motion capture systems, it may be suggested that 3D-MCS present practitioners with an opportunity to reliably and efficiently measure the movement characteristics of patients and athletes. This has implications for monitoring the health/performance of a broad range of populations.

Relationship between Upper and Lower Body Strength and Basketball Shooting Performance.

Strength is one of the key physiological performance attributes related to optimal on-court basketball performance. However, there is a lack of scientific literature studying how strength relates to shooting proficiency, as a key basketball skill capable of discriminating winning from losing game outcomes. Thus, the purpose of the present study was to examine the relationship between maximal upper and lower body strength and free-throw, two-point, and three-point shooting accuracy. Ten males and seven females performed bench press and back squat one repetition maximum (1RM) and basketball shooting testing during two laboratory visits. The shooting protocol consisted of five sets of 15 free-throw, two-point, and three-point shots performed in sequential order. Each set was separated by a 30 min rest interval to minimize the influence of fatigue. Each subject attempted 225 shots, combining for a total of 3825 shots. The average free-throw, two-point, and three-point shooting accuracy for men were 74.5 ± 11.9, 68.4 ± 9.9, and 53.3 ± 14.9%, and for women 79.2 ± 11.2, 65.5 ± 8.4, and 51.2 ± 15.3%, respectively. The average bench press and back squat 1RM for men was 88.2 ± 18.6 and 117.0 ± 21.2 kg, and for women, 40.6 ± 7.5 and 66.9 ± 9.9 kg, respectively. The findings of the present study revealed no significant relationships between maximal upper and lower body strength and basketball shooting performance for both male and female participants. Neither bench press nor back squat 1RM was a good predictor of free-throw, two-point, and three-point shooting performance.

Impact of Distance and Proficiency on Shooting Kinematics in Professional Male Basketball Players.

Shooting efficiency is one of the key performance parameters related to securing the desired game outcome at various levels of basketball competition, and it is largely influenced by the biomechanical adjustments incorporated during the preparatory and release phase of the shooting motion. Thus, the purpose of the present study was twofold: (a) to examine the differences in the kinematic characteristics between free-throw, two-point, and three-point shots, and (b) to examine the differences between shooters with excellent (≥80%) and good (<80%) levels of shooting proficiency. A total of 10 professional male basketball players performed 5 free-throw (4.57 m), two-point (5.18 m), and three-point (6.75 m) shots, combining for a total of 150 shots. A high-definition camera recording at 120 fps was used to capture the shooting motion from a sagittal point of view, and video analysis software was used to analyze the kinematic variables of interest. The findings of the present study reveal that the kinematic characteristics during the preparatory phase of the shooting motion remain unchanged between free-throw and two-point shots. Three-point shots required lower elbow positioning, influenced by greater knee and hip flexion when compared to free-throw and two-point shots. The release angle was notably lower for shots attempted beyond the three-point line but remained unchanged between the free-throw and two-point shooting motions. Release height and vertical displacement were significantly greater for two- and three-point shots when compared to free-throw shots, while no difference was observed between the two- and three-point shots. In addition, no significant differences in shooting kinematics were observed between those participants with excellent and good levels of shooting proficiency.

High-Precision Triple Oxygen Isotope Analysis of Carbon Dioxide by Tunable Infrared Laser Absorption Spectroscopy.

Precision measurements of the stable isotope ratios of oxygen (18O/16O and 17O/16O) in CO2 are critical to atmospheric monitoring and terrestrial climate research. High-precision 17O measurements by isotope ratio mass spectrometry (IRMS) are challenging because they require complicated sample preparation procedures, long measurement times, and relatively large samples sizes. Recently, tunable infrared laser direct absorption spectroscopy (TILDAS) has shown significant potential as an alternative technique for triple oxygen isotope analysis of CO2, although the ultimate level of reproducibility is unknown, partly because it is unclear how to relate TILDAS measurements to an internationally accepted isotope abundance scale (e.g., VSMOW2-SLAP2). Here, we present a method for high-precision triple oxygen isotope analysis of CO2 by TILDAS, requiring ∼8-9 µmol of CO2 (or 0.9 mg carbonate) in 50 min, plus ∼1.5 h for sample preparation and dilution of CO2 in N2 to a nominal 400 µmol mol-1. Overall reproducibility of Δ'17O (CO2) was 0.004‰ (4 per meg) for IAEA603 (SE, n = 6) and 10 per meg for NBS18 (SE, n = 4). Values corrected to the VSMOW2-SLAP2 scale are in good agreement with established techniques of high-precision IRMS, with the exception of Δ'17O measured by platinum-catalyzed exchange of CO2 with O2. Compared to high-precision IRMS, TILDAS offers the advantage of ∼10 times less sample, and greater throughput, without loss of reproducibility. The flexibility of the technique should allow for many important applications to global biogeochemical monitoring and investigation of 17O anomalies in a range of geological materials.

A WC/WO star exploding within an expanding carbon-oxygen-neon nebula.

The final fate of massive stars, and the nature of the compact remnants they leave behind (black holes and neutron stars), are open questions in astrophysics. Many massive stars are stripped of their outer hydrogen envelopes as they evolve. Such Wolf-Rayet stars1 emit strong and rapidly expanding winds with speeds greater than 1,000 kilometres per second. A fraction of this population is also helium-depleted, with spectra dominated by highly ionized emission lines of carbon and oxygen (types WC/WO). Evidence indicates that the most commonly observed supernova explosions that lack hydrogen and helium (types Ib/Ic) cannot result from massive WC/WO stars2,3, leading some to suggest that most such stars collapse directly into black holes without a visible supernova explosion4. Here we report observations of SN 2019hgp, beginning about a day after the explosion. Its short rise time and rapid decline place it among an emerging population of rapidly evolving transients5-8. Spectroscopy reveals a rich set of emission lines indicating that the explosion occurred within a nebula composed of carbon, oxygen and neon. Narrow absorption features show that this material is expanding at high velocities (greater than 1,500 kilometres per second), requiring a compact progenitor. Our observations are consistent with an explosion of a massive WC/WO star, and suggest that massive Wolf-Rayet stars may be the progenitors of some rapidly evolving transients.

Return of Results Policies for Genomic Research: Current Practices and the Hearts in Rhythm Organization (HiRO) Approach.

Research teams developing biobanks and/or genomic databases must develop policies for the disclosure and reporting of potentially actionable genomic results to research participants. Currently, a broad range of approaches to the return of results exist, with some studies opting for nondisclosure of research results and others following clinical guidelines for the return of potentially actionable findings from sequencing. In this review, we describe current practices and highlight decisions a research team must make when designing a return of results policy, from informed consent to disclosure practices and clinical validation options. The unique challenges of returning incidental findings in cardiac genes, including reduced penetrance and the lack of clinical screening standards for phenotype-negative individuals, are discussed. Finally, the National Hearts in Rhythm Organisation (HiRO) Registry approach is described to provide a rationale for the selective return of field-specific variants to those participating in disease-specific research. Our goal is to provide researchers with a resource when developing a return of results policy tailored for their research program, based on unique factors related to study design, research team composition, and availability of clinical resources.

Contributions of side effects to contraceptive discontinuation and method switch among Kenyan women: a prospective cohort study.

OBJECTIVE: To determine the contribution of specific contraceptive side effects to method switch and modern-method discontinuation among Kenyan women. DESIGN: A prospective cohort study. SETTING: Five counties in Western Kenya. PARTICIPANTS: Women aged ≥18 years old and emancipated female minors ≥14 years old using modern, reversible contraception were recruited while attending 10 public health facilities. METHODS: Patient-reported adverse effect symptoms, method switch and discontinuation were reported through weekly text message-based surveys for 24 weeks. MAIN OUTCOME MEASUREMENTS: Prevalence, hazards ratio (HR). RESULTS: Among 825 women, 44% were using implants, 43% injectables, 7% an intrauterine device and 6% oral contraceptive pills at enrolment. Most (61%) women were continuing a method used in the previous month. During the 24-week follow up, incidence of contraceptive switch was 61.3 per 100 person-years (95% confidence interval [CI] 52.4-71.8) and incidence of discontinuation was 38.5 per 100 person-years (95% CI 31.6-47.0). On average, one-quarter (prevalence [Pr] 0.24, 95% CI 0.22-0.26) of participants reported side effects or method problems weekly, with sexual side effects the most prevalent symptom (Pr 0.15, 95% CI 0.13-0.16). Lack of expected bleeding was associated with higher risk of method switch (adjusted hazard ratio [aHR] 2.36, 95% CI 1.22-4.57). Risk of all-modern method discontinuation was higher among women experiencing irregular bleeding (aHR 2.39, 95% CI 1.20-4.77), weight changes (aHR 2.72, 95% CI 1.47-4.68) and sexual side effects (aHR 2.42, 95% CI 1.40-4.20). CONCLUSIONS: Addressing irregular bleeding, weight changes and sexual side effects through development of new products that minimise these specific side effects and anticipatory counseling may reduce method-related discontinuation. TWEETABLE ABSTRACT: Bleeding, weight changes, sexual problems associated with discontinuation of #contraception, but many continue despite side effects.

Arrhythmic Mitral Valve Prolapse and Mitral Annulus Disjunction in Heritable Aortic Disease.

Patients with heritable aortic disease (HAD) have an increased risk of ventricular arrhythmias and sudden cardiac death. Although mitral valve prolapse is common in HAD, mitral annulus disjunction (MAD) has only recently been described in these patients. This under-recognized condition may be a contributing factor to otherwise unexplained ventricular arrhythmias and sudden cardiac death in patients with HAD. This case series describes 3 patients in an adult HAD clinic who have concomitant mitral valve prolapse, MAD, and malignant arrhythmias. These cases may represent a unique disease entity or overlap syndrome, and they introduce MAD as a potential arrhythmogenic risk marker in HAD.

Les patients atteints de maladie aortique héréditaire (MAH) présentent un risque accru d'arythmie ventriculaire et de mort subite d'origine cardiaque. Bien que le prolapsus valvulaire mitral soit fréquent dans les cas de MAH, la disjonction annulaire mitrale (DAM) n'a été décrite que récemment chez ces patients. Cet état méconnu peut être un facteur contribuant à des arythmies ventriculaires autrement inexpliquées et à la mort subite d'origine cardiaque chez les patients atteints de MAH. Cette série de cas décrit trois patients d'une clinique de MAH pour adultes qui présentent un prolapsus valvulaire mitral, une DAM et des arythmies malignes en concomitance. Ces cas peuvent représenter une entité morbide unique ou un syndrome de chevauchement, et laissent entendre que la DAM pourrait être un nouveau marqueur du risque arythmogène associé à la MAH.

Context Matters: Social Psychological Factors That Underlie Academic Performance across Seven Institutions.

To enhance equity and diversity in undergraduate biology, recent research in biology education focuses on best practices that reduce learning barriers for all students and improve academic performance. However, the majority of current research into student experiences in introductory biology takes place at large, predominantly White institutions. To foster contextual knowledge in biology education research, we harnessed data from a large research coordination network to examine the extent of academic performance gaps based on demographic status across institutional contexts and how two psychological factors, test anxiety and ethnicity stigma consciousness, may mediate performance in introductory biology. We used data from seven institutions across three institution types: 2-year community colleges, 4-year inclusive institutions (based on admissions selectivity; hereafter, inclusive), and 4-year selective institutions (hereafter, selective). In our sample, we did not observe binary gender gaps across institutional contexts, but found that performance gaps based on underrepresented minority status were evident at inclusive and selective 4-year institutions, but not at community colleges. Differences in social psychological factors and their impacts on academic performance varied substantially across institutional contexts. Our findings demonstrate that institutional context can play an important role in the mechanisms underlying performance gaps.

Rapid and inexpensive preparation of genome-wide nucleosome footprints from model and non-model organisms.

MNase-seq (micrococcal nuclease sequencing) is used to map nucleosome positions in eukaryotic genomes to study the relationship between chromatin structure and DNA-dependent processes. Current protocols require at least two days to isolate nucleosome-protected DNA fragments. We have developed a streamlined protocol for S. cerevisiae and other fungi which takes only three hours. Modified protocols were developed for wild fungi and mammalian cells. This method for rapidly producing sequencing-ready nucleosome footprints from several organisms makes MNase-seq faster and easier, with less chemical waste.

Human METTL7B is an alkyl thiol methyltransferase that metabolizes hydrogen sulfide and captopril.

Methylation of alkyl thiols is a biotransformation pathway designed to reduce thiol reactivity and potential toxicity, yet the gene and protein responsible for human alkyl thiol methyltransferase (TMT) activity remain unknown. Here we demonstrate with a range of experimental approaches using cell lines, in vitro systems, and recombinantly expressed enzyme, that human methyltransferase-like protein 7B (METTL7B) catalyzes the transfer of a methyl group from S-adenosyl-L-methionine (AdoMet) to hydrogen sulfide (H2S) and other exogenous thiol small molecules. METTL7B gene modulation experiments, including knockdown in HepG2 cells and overexpression in HeLa cells, directly alter the methylation of the drug captopril, a historic probe substrate for TMT activity. Furthermore, recombinantly expressed and purified wild-type METTL7B methylates several thiol compounds, including H2S, 7α-thiospironolactone, L-penicillamine, and captopril, in a time- and concentration-dependent manner. Typical for AdoMet-dependent small molecule methyltransferases, S-adenosyl-L-homocysteine (AdoHcy) inhibited METTL7B activity in a competitive fashion. Similarly, mutating a conserved aspartate residue, proposed to anchor AdoMet into the active site, to an alanine (D98A) abolished methylation activity. Endogenous thiols such as glutathione and cysteine, or classic substrates for other known small molecule S-, N-, and O-methyltransferases, were not substrates for METTL7B. Our results confirm, for the first time, that METTL7B, a gene implicated in multiple disease states including rheumatoid arthritis and breast cancer, encodes a protein that methylates small molecule alkyl thiols. Identifying the catalytic function of METTL7B will enable future pharmacological research in disease pathophysiology where altered METTL7B expression and, potentially H2S levels, can disrupt cell growth and redox state.

Basis of specificity for a conserved and promiscuous chromatin remodeling protein.

Eukaryotic genomes are organized dynamically through the repositioning of nucleosomes. Isw2 is an enzyme that has been previously defined as a genome-wide, nonspecific nucleosome spacing factor. Here, we show that Isw2 instead acts as an obligately targeted nucleosome remodeler in vivo through physical interactions with sequence-specific factors. We demonstrate that Isw2-recruiting factors use small and previously uncharacterized epitopes, which direct Isw2 activity through highly conserved acidic residues in the Isw2 accessory protein Itc1. This interaction orients Isw2 on target nucleosomes, allowing for precise nucleosome positioning at targeted loci. Finally, we show that these critical acidic residues have been lost in the Drosophila lineage, potentially explaining the inconsistently characterized function of Isw2-like proteins. Altogether, these data suggest an 'interacting barrier model,' where Isw2 interacts with a sequence-specific factor to accurately and reproducibly position a single, targeted nucleosome to define the precise border of phased chromatin arrays.

DNA encodes the genetic instructions for life in a long, flexible molecular chain that is packaged up neatly to fit inside cells. Short sections of DNA are wound around proteins to form bundles called nucleosomes, and then spun into chromatin fibres, a more compact form of DNA. While nucleosomes are a fundamental part of this space-saving packaging process, they also play a key regulatory role in gene expression, which is where genes are decoded into working proteins. Placing nucleosomes at regular intervals along DNA invariably controls which parts of the DNA ­ and which genes ­ the cell's machinery can access and 'read' to make proteins. But the nucleosomes' positions are not fixed, and gene expression is a dynamic process. The cell often uncoils and repackages its DNA while molecular motors called chromatin remodelling proteins move nucleosomes up and down the DNA, exposing some genes and obstructing others. One group of chromatin remodelling proteins are called Imitation Switch (ISWI) complexes. It has long been thought that these complexes position nucleosomes with little regard to the underlying DNA sequence or the genes encoded, that is to say in a non-specific way. However, this theory has not been thoroughly tested. It is possible that ISWI complexes actually place nucleosomes on certain parts of DNA at particular times in an organism's development, or in response to other environmental factors. Except how such precision is achieved remains unknown. To test this alternative theory of nucleosome positioning, Donovan et al. studied ISWI proteins and nucleosomes in common baker's yeast. This involved systematically removing sections of ISWI proteins to see whether the complexes could still position nucleosomes, and which parts of the proteins where essential for the job. By doing so, Donovan et al. identified multiple 'targeting' proteins that bind to ISWI proteins and deliver the complexes to specific target sequences of DNA. From there, the complex remodels the nucleosome, positioning it at a specific distance from its landing site on DNA, as further experiments showed. This research provides a new model for explaining how nucleosomes are positioned to package DNA and control gene expression. Donovan et al. have identified a new mechanism of interaction between nucleosomes and chromatin remodelling proteins of the ISWI variety. It is possible that more interactions of this kind will be discovered with further research.