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Double-strand breaks (DSBs) are the most deleterious lesions experienced by our genome. Yet, DSBs are intentionally induced during gamete formation to promote the exchange of genetic material between homologous chromosomes. While the conserved topoisomerase-like enzyme Spo11 catalyzes DSBs, additional regulatory proteins-referred to as "Spo11 accessory factors"- regulate the number, timing, and placement of DSBs during early meiotic prophase ensuring that SPO11 does not wreak havoc on the genome. Despite the importance of the accessory factors, they are poorly conserved at the sequence level suggesting that these factors may adopt unique functions in different species. In this work, we present a detailed analysis of the genetic and physical interactions between the DSB factors in the nematode Caenorhabditis elegans providing new insights into conserved and novel functions of these proteins. This work shows that HIM-5 is the determinant of X-chromosome-specific crossovers and that its retention in the nucleus is dependent on DSB-1, the sole accessory factor that interacts with SPO-11. We further provide evidence that HIM-5 coordinates the actions of the different accessory factors sub-groups, providing insights into how components on the DNA loops may interact with the chromosome axis.

Background: The Health E Englewood Health and Wellness Program is a social determinant of health (SDoH) intervention developed to address social factors affecting the health of the North Hudson Community Action Corporation (NHCAC) patients', a Federally Qualified Health Center located in Englewood, New Jersey. The main aim of this integrated wellness approach was to educate and motivate participants from the local community by strengthening the development of healthy lifestyles and providing the necessary tools for positive behavior change., Methods: Health E Englewood was a four consecutive week workshop series focused on three areas of health: physical, emotional, and nutritional wellness. The program targeted Spanish-speaking patients from NHCAC and was offered virtually via Zoom in Spanish., Results: The Health E Englewood program was launched in October 2021 with 40 active participants. About 63% of participants attended at least three of the four workshop sessions, with at least 60% of participants reporting improved lifestyle changes after the program. Additional follow-up data collected six months later also indicated evidence of the program's long-term benefits., Discussion: Social factors are the primary drivers of health outcomes. While many determinant interventions have failed to show long-lasting benefits, studying these interventions and their impact is crucial as it avoids "re-creating the wheel" inside health care and increasing costs., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2023, Brijmohan et al.)

Male infertility impacts millions of couples yet, the etiology of primary infertility remains largely unknown. A critical element of successful spermatogenesis is maintenance of genome integrity. Here, we present a genomic study of spermatogenic failure (SPGF). Our initial analysis (n = 176) did not reveal known gene-candidates but identified a potentially significant single-nucleotide variant (SNV) in X-linked germ-cell nuclear antigen (GCNA). Together with a larger follow-up study (n = 2049), 7 likely clinically relevant GCNA variants were identified. GCNA is critical for genome integrity in male meiosis and knockout models exhibit impaired spermatogenesis and infertility. Single-cell RNA-seq and immunohistochemistry confirm human GCNA expression from spermatogonia to elongated spermatids. Five identified SNVs were located in key functional regions, including N-terminal SUMO-interacting motif and C-terminal Spartan-like protease domain. Notably, variant p.Ala115ProfsTer7 results in an early frameshift, while Spartan-like domain missense variants p.Ser659Trp and p.Arg664Cys change conserved residues, likely affecting 3D structure. For variants within GCNA's intrinsically disordered region, we performed computational modeling for consensus motifs. Two SNVs were predicted to impact the structure of these consensus motifs. All identified variants have an extremely low minor allele frequency in the general population and 6 of 7 were not detected in > 5000 biological fathers. Considering evidence from animal models, germ-cell-specific expression, 3D modeling, and computational predictions for SNVs, we propose that identified GCNA variants disrupt structure and function of the respective protein domains, ultimately arresting germ-cell division. To our knowledge, this is the first study implicating GCNA, a key genome integrity factor, in human male infertility.

Objectives: To investigate the incidence of chromosomal abnormalities in the products of conception (POC) of patients with spontaneous miscarriages (SM) and with recurrent pregnancy losses (RPL) and to determine biological mechanisms contributing to RPL., Methods: During a 20-year period, 12 096 POC samples underwent classical chromosome analysis. Cytogenetic findings were compared between the SM and RPL cohorts., Results: Analysis of RPL cohort has identified an increased incidence of inherited and de novo structural chromosome abnormalities, recurrent polyploid conceptions, and complex mosaic alterations. These abnormalities are the signature of genomic instability, posing a high risk of genetic abnormalities to offspring independent of maternal age. Predominance of male conceptions in the RPL cohort points toward an X-linked etiology and gender-specific intolerance for certain genetic abnormalities., Conclusions: Our study showed several possible genetic etiologies of RPL, including parental structural chromosome rearrangements, predisposition to meiotic nondisjunction, and genomic instability. Loss of karyotypically normal fetuses might be attributed to defects in genes essential for fetal development, as well as aberrations affecting the X chromosome. Molecular studies of parental and POC genomes will help to identify inherited defects in genes involved in meiotic divisions and DNA repair to confirm our hypotheses, and to discover novel fetal-essential genes., (© 2020 John Wiley & Sons Ltd.)

Elimination of the proliferating germline extends lifespan in C. elegans. This phenomenon provides a unique platform to understand how complex metazoans retain metabolic homeostasis when challenged with major physiological perturbations. Here, we demonstrate that two conserved transcription regulators essential for the longevity of germline-less adults, DAF-16/FOXO3A and TCER-1/TCERG1, concurrently enhance the expression of multiple genes involved in lipid synthesis and breakdown, and that both gene classes promote longevity. Lipidomic analyses revealed that key lipogenic processes, including de novo fatty acid synthesis, triglyceride production, desaturation and elongation, are augmented upon germline removal. Our data suggest that lipid anabolic and catabolic pathways are coordinately augmented in response to germline loss, and this metabolic shift helps preserve lipid homeostasis. DAF-16 and TCER-1 also perform essential inhibitory functions in germline-ablated animals. TCER-1 inhibits the somatic gene-expression program that facilitates reproduction and represses anti-longevity genes, whereas DAF-16 impedes ribosome biogenesis. Additionally, we discovered that TCER-1 is critical for optimal fertility in normal adults, suggesting that the protein acts as a switch supporting reproductive fitness or longevity depending on the presence or absence of the germline. Collectively, our data offer insights into how organisms adapt to changes in reproductive status, by utilizing the activating and repressive functions of transcription factors and coordinating fat production and degradation.

Maintenance of the genome requires the continual repair of DNA lesions. Exposure of nematodes to DNA damage-inducing agents is a powerful method to rapidly ascribe a role for specific genes in DNA repair and to define epistatic relationships to other repair genes which allows for the construction of repair pathways. Despite the extensive use of these agents, however, differences in dosing, timing, and handling makes it difficult to compare results across laboratories. We provide herein a consideration of the parameters that influence the results of these exposures and detailed protocols for the exposure to mutagenic inducing agents., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Certification gasoline was splash blended with alcohols to produce four blends: ethanol (16 vol%), n-butanol (17 vol%), i-butanol (21 vol%), and an i-butanol (12 vol%)/ethanol (7 vol%) mixture; these fuels were tested in a 2009 Honda Odyssey (a Tier 2 Bin 5 vehicle) over triplicate LA92 cycles. Emissions of oxides of nitrogen, carbon monoxide, non-methane organic gases (NMOG), unburned alcohols, carbonyls, and C1-C8 hydrocarbons (particularly 1,3-butadiene and benzene) were determined. Large, statistically significant fuel effects on regulated emissions were a 29% reduction in CO from E16 and a 60% increase in formaldehyde emissions from i-butanol, compared to certification gasoline. Ethanol produced the highest unburned alcohol emissions of 1.38 mg/mile ethanol, while butanols produced much lower unburned alcohol emissions (0.17 mg/mile n-butanol, and 0.30 mg/mile i-butanol); these reductions were offset by higher emissions of carbonyls. Formaldehyde, acetaldehyde, and butyraldehyde were the most significant carbonyls from the n-butanol blend, while formaldehyde, acetone, and 2-methylpropanal were the most significant from the i-butanol blend. The 12% i-butanol/7% ethanol blend was designed to produce no increase in gasoline vapor pressure. This fuel's exhaust emissions contained the lowest total oxygenates among the alcohol blends and the lowest NMOG of all fuels tested.

Nine flex-fuel vehicles meeting Tier 1, light duty vehicle-low emission vehicle (LDV-LEV), light duty truck 2-LEV (LDT2-LEV), and Tier 2 emission standards were tested over hot-start and cold-start three-phase LA92 cycles for nonmethane organic gases, ethanol, acetaldehyde, formaldehyde, acetone, nitrous oxide, nitrogen oxides (NO(x)), carbon monoxide (CO), and carbon dioxide (CO(2)), as well as fuel economy. Emissions were measured immediately after refueling with E40. The vehicles had previously been adapted to either E10 or E76. An overall comparison of emissions and fuel economy behavior of vehicles running on E40 showed results generally consistent with adaptation to the blend after the length of the three-phase hot-start LA92 test procedure (1735 s, 11 miles). However, the single LDT2-LEV vehicle, a Dodge Caravan, continued to exhibit statistically significant differences in emissions for most pollutants when tested on E40 depending on whether the vehicle had been previously adapted to E10 or E76. The results were consistent with an overestimate of the amount of ethanol in the fuel when E40 was added immediately after the use of E76. Increasing ethanol concentration in fuel led to reductions in fuel economy, NO(x), CO, CO(2), and acetone emissions as well as increases in emissions of ethanol, acetaldehyde, and formaldehyde.

The perpetuation of most eukaryotic species requires differentiation of pluripotent progenitors into egg and sperm and subsequent fusion of these gametes to form a new zygote. Meiosis is a distinguishing feature of gamete formation as it leads to the twofold reduction in chromosome number thereby maintaining ploidy across generations. This process increases offspring diversity through the random segregation of chromosomes and the exchange of genetic material between homologous parental chromosomes, known as meiotic crossover recombination. These exchanges require the establishment of unique and dynamic chromatin configurations that facilitate cohesion, homolog pairing, synapsis, double strand break formation and repair. The precise orchestration of these events is critical for gamete survival as demonstrated by the majority of human aneuploidies that can be traced to defects in the first meiotic division (Hassold T, Hall H, Hunt P: The origin of human aneuploidy: where we have been, where we are going. Hum Mol Genet 2007, 16 Spec No. 2:R203-R208.). This review will focus on recent advances in our understanding of key meiotic events and how coordination of these events is occurring., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

E85, which consists of nominally 85% fuel grade ethanol and 15% gasoline, must be used in flexible-fuel (or "flex-fuel") vehicles (FFVs) that can operate on fuel with an ethanol content of 0-85%. Published studies include measurements of the effect of E85 on tailpipe emissions for Tier 1 and older vehicles. Car manufacturers have also supplied a large body of FFV certification data to the U.S. Environmental Protection Agency, primarily on Tier 2 vehicles. These studies and certification data reveal wide variability in the effects of E85 on emissions from different vehicles. Comparing Tier 1 FFVs running on E85 to similar non-FFVs running on gasoline showed, on average, significant reductions in emissions of oxides of nitrogen (NO x ; 54%), non-methane hydrocarbons (NMHCs; 27%), and carbon monoxide (CO; 18%) for E85. Comparing Tier 2 FFVs running on E85 and comparable non-FFVs running on gasoline shows, for E85 on average, a signifi-cant reduction in emissions of CO (20%), and no signifi-cant effect on emissions of non-methane organic gases (NMOGs). NO x emissions from Tier 2 FFVs averaged approximately 28% less than comparable non-FFVs. However, perhaps because of the wide range of Tier 2 NO x standards, the absolute difference in NO x emissions between Tier 2 FFVs and non-FFVs is not significant (P =0.28). It is interesting that Tier 2 FFVs operating on gasoline produced approximately 13% less NMOGs than non-FFVs operating on gasoline. The data for Tier 1 vehicles show that E85 will cause significant reductions in emissions of benzene and butadiene, and significant increases in emissions of formaldehyde and acetaldehyde, in comparison to emissions from gasoline in both FFVs and non-FFVs. The compound that makes up the largest proportion of organic emissions from E85-fueled FFVs is ethanol.

Homologous recombination (HR) plays a critical role in the restart of blocked replication forks, but how this is achieved remains poorly understood. We show that mutants in the single Rad51 paralog in Caenorhabditis elegans, rfs-1, permit discrimination between HR substrates generated at DNA double-strand breaks (DSBs), or following replication fork collapse from HR substrates assembled at replication fork barriers (RFBs). Unexpectedly, RFS-1 is dispensable for RAD-51 recruitment to meiotic and ionizing radiation (IR)-induced DSBs and following replication fork collapse, yet, is essential for RAD-51 recruitment to RFBs formed by DNA crosslinking agents and other replication blocking lesions. Deletion of rfs-1 also suppresses the accumulation of toxic HR intermediates in him-6; top-3 mutants and accelerates deletion formation at presumed endogenous RFBs formed by poly G/C tracts in the absence of DOG-1. These data suggest that RFS-1 is not a general mediator of HR-dependent DSB repair, but acts specifically to promote HR at RFBs. HR substrates generated at conventional DSBs or following replication fork collapse are therefore intrinsically different from those produced during normal repair of blocked replication forks.

The combination of a powerful well-designed user interface with detailed high-quality data sets can create new possibilities for data exploration and analysis. The Allen Brain Atlas (http://www.brain-map.org) provides a collection of tools for examining a set of images that detail gene expression in the mouse brain. Powerful web-based viewers for individual images and parallel examination of related images interact with an external application for three-dimensional views. The underlying dataset, generated via high-throughput analysis of expression patterns of more than 21,000 genes in adult mouse brains, provides three-dimensional views of gene expression patterns displayed in the context of an anatomical ontology. Facilities for filtering views, saving views of interest, annotating images and sharing views via email support the ongoing process of analysis and provide a model for the future of integrated tools for analysing large image data sets.

In C. elegans, the Sma/Mab TGFbeta signaling pathway regulates body size and male tail patterning. SMA-9, the C. elegans homolog of Schnurri, has been shown to function as a downstream component to mediate the Sma/Mab TGFbeta signaling pathway in these processes. We have discovered a new role for SMA-9 in dorsoventral patterning of the C. elegans post-embryonic mesoderm, the M lineage. In addition to a small body size, sma-9 mutant animals exhibit a dorsal-to-ventral fate transformation within the M lineage. This M lineage defect of sma-9 mutants is unique in that animals carrying mutations in all other known components of the TGFbeta pathway exhibit no M lineage defects. Surprisingly, mutations in the core components of the Sma/Mab TGFbeta signaling pathway suppressed the M lineage defects of sma-9 mutants without suppressing their body size defects. We show that this suppression specifically happens within the M lineage. Our studies have uncovered an unexpected role of SMA-9 in antagonizing the TGFbeta signaling pathway during mesodermal patterning, suggesting a novel mode of function for the SMA-9/Schnurri family of proteins.

During development, progression through the cell cycle must be coordinately regulated with cellular differentiation. Despite significant progress in identifying genes required independently for each of these processes, the molecules which facilitate this cross talk have for the most part been elusive. Using the six macrophage-like coelomocytes of the nematode Caenorhabditis elegans as a model system to gain insight into the mesodermal differentiation pathway, we have isolated a set of mutants that alter coelomocyte numbers. One of these mutations, cc600, apparently results from a partial loss-of-function in the C. elegans cyclin D gene, cyd-1. The mutant has coelomocyte-specific defects without changes in other lineages. The mutants show that cell growth, terminal differentiation and cellular function proceed in the absence of cyd-1 activity and cell division. The results suggest that certain mesodermal lineages may be uniquely affected by changes in cyd-1 activity.

Certification testing of locomotive diesel engines is conducted by testing the locomotive in a series of steady-state conditions, or notches. The aim of this work was to determine whether notch changes, which are made on the order of twice per minute during normal locomotive operation, significantly affect emission rates. Particulate matter (PM) measurements recorded by others over 5, 15, and 30 min immediately at notch change were analyzed. By assuming that PM emissions during steady-state conditions were constant, it was possible to determine the amount of PM emitted because of notch change. Certification line-haul and switching duty cycles were modified to include a representative number of notch changes. The results of these calculations suggest that in test cycles in which a representative number of notch changes were included, approximately 40% of PM emissions occurred because of notch changes.

A model of a heavy-duty vehicle driveline with automatic transmission has been developed for estimating engine speed and load from vehicle speed. The model has been validated using emissions tests conducted on three diesel vehicles on a chassis dynamometer and then on the engines removed from the vehicles tested on an engine dynamometer. Nitrogen oxide (NOx) emissions were proportional to work done by the engine. For two of the engines, the NOx/horsepower(HP) ratio was the same on the engine and on the chassis dynamometer tests. For the third engine NOx/HP was significantly higher from the chassis test, possibly due to the use of dual engine maps. The engine certification test generated consistently less particulate matter emissions on a gram per brake horsepower-hour basis than the Heavy Duty Transient and Central Business District chassis cycles. A good linear correlation (r2 = 0.97 and 0.91) was found between rates of HP increase integrated over the test cycle and PM emissions for both the chassis and the engine tests for two of the vehicles. The model also shows how small changes in vehicle speeds can lead to a doubling of load on the engine. Additionally, the model showed that it is impossible to drive a vehicle cycle equivalent to the heavy-duty engine federal test procedure on these vehicles.

Idle emissions of total hydrocarbon (THC), CO, NOx, and particulate matter (PM) were measured from 24 heavy-duty diesel-fueled (12 trucks and 12 buses) and 4 heavy-duty compressed natural gas (CNG)-fueled vehicles. The volatile organic fraction (VOF) of PM and aldehyde emissions were also measured for many of the diesel vehicles. Experiments were conducted at 1609 m above sea level using a full exhaust flow dilution tunnel method identical to that used for heavy-duty engine Federal Test Procedure (FTP) testing. Diesel trucks averaged 0.170 g/min THC, 1.183 g/min CO, 1.416 g/min NOx, and 0.030 g/min PM. Diesel buses averaged 0.137 g/min THC, 1.326 g/min CO, 2.015 g/min NOx, and 0.048 g/min PM. Results are compared to idle emission factors from the MOBILE5 and PART5 inventory models. The models significantly (45-75%) overestimate emissions of THC and CO in comparison with results measured from the fleet of vehicles examined in this study. Measured NOx emissions were significantly higher (30-100%) than model predictions. For the pre-1999 (pre-consent decree) truck engines examined in this study, idle NOx emissions increased with model year with a linear fit (r2 = 0.6). PART5 nationwide fleet average emissions are within 1 order of magnitude of emissions for the group of vehicles tested in this study. Aldehyde emissions for bus idling averaged 6 mg/min. The VOF averaged 19% of total PM for buses and 49% for trucks. CNG vehicle idle emissions averaged 1.435 g/min for THC, 1.119 g/min for CO, 0.267 g/min for NOx, and 0.003 g/min for PM. The g/min PM emissions are only a small fraction of g/min PM emissions during vehicle driving. However, idle emissions of NOx, CO, and THC are significant in comparison with driving emissions.

It has previously been shown that germ cells in embryos derived from nos mutant mothers do not migrate to the primitive gonad and prematurely express several germline-specific markers. In the studies reported here, we have traced these defects back to the syncytial blastoderm stage. We show that pole cells in nos embryos fail to establish/maintain transcriptional quiescence; the sex determination gene Sex-lethal (Sxl) and the segmentation genes fushi tarazu and even-skipped are ectopically activated in nos- germ cells. We show that nos- germ cells are unable to attenuate the cell cycle and instead continue dividing. Unexpectedly, removal of the Sxl gene in the zygote mitigates both the migration and mitotic defects of nos- germ cells. Supporting the conclusion that Sxl is an important target for nos repression, ectopic, premature expression of Sxl protein in germ cells disrupts migration and stimulates mitotic activity.

In Drosophila melanogaster, Sex-lethal (Sxl) controls autoregulation and sexual differentiation by alternative splicing but regulates dosage compensation by translational repression. To elucidate how Sxl functions in splicing and translational regulation, we have ectopically expressed a full-length Sxl protein (Sx.FL) and a protein lacking the N-terminal 40 amino acids (Sx-N). The Sx.FL protein recapitulates the activity of Sxl gain-of-function mutations, as it is both sex transforming and lethal in males. In contrast, the Sx-N protein unlinks the sex-transforming and male-lethal effects of Sxl. The Sx-N proteins are compromised in splicing functions required for sexual differentiation, displaying only partial autoregulatory activity and almost no sex-transforming activity. On the other hand, the Sx-N protein does retain substantial dosage compensation function and kills males almost as effectively as the Sx.FL protein. In the course of our analysis of the Sx.FL and Sx-N transgenes, we have also uncovered a novel, negative autoregulatory activity, in which Sxl proteins bind to the 3' untranslated region of Sxl mRNAs and decrease Sxl protein expression. This negative autoregulatory activity may be a homeostasis mechanism.