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The objective of this study was to evaluate and to estimate the relationship between fecal egg counts (FECs) and FAMACHA score and the body weight of growing Katahdin rams during a parasite challenge. One of the largest factors negatively influencing reproduction and economics in the sheep industry is gastrointestinal nematode (GIN) parasites. Due to anthelmintic resistance of these parasites, animals are selected for parasite resistance using FEC and FAMACHA scores. Data were used from the Virginia Tech Southwest Agricultural Research and Extension Center Ram Test in Glade Spring, VA, from the year 2012 to 2018 in which animals were tested in 14-d intervals for 70 d. Mixed models for repeated weight measurements were made from backward stepwise selection to evaluate the relationships between weight and GIN FEC. A total of 576 animals within 23 contemporary groups derived from test year and pasture group were analyzed. Ram, contemporary group, and consignor were considered random effects, and fixed effects were birth type, test day, age, age squared, starting weight, FEC, and FAMACHA score. Pairwise contrasts were used in the statistical analysis of parameters and their interactions. Weight and age were found to have a quadratic relationship. Increased FEC was associated with weight loss at a rate of 0.00030 kg/FEC ( P < 0.0001). Animals dewormed at any point during the trial weighed less than those that were not and increased with test day to a maximum difference of 4.66 kg ( P < 0.001). FAMACHA score was found to be significant ( P < 0.05), but a direct relationship with weight was not conclusive. Overall, rams with severe enough parasite load to require deworming had lesser weights, which could impact the profitability of sheep production and reinforced the need to select animals that had greater innate parasite resistance., (© The Author(s) 2020. Published by Oxford University Press on behalf of the American Society of Animal Science.)

Identifying stabilising variants of membrane protein targets is often required for structure determination. Our new computational pipeline, the Integral Membrane Protein Stability Selector (IMPROvER) provides a rational approach to variant selection by employing three independent approaches: deep-sequence, model-based and data-driven. In silico tests using known stability data, and in vitro tests using three membrane protein targets with 7, 11 and 16 transmembrane helices provided measures of success. In vitro, individual approaches alone all identified stabilising variants at a rate better than expected by random selection. Low numbers of overlapping predictions between approaches meant a greater success rate was achieved (fourfold better than random) when approaches were combined and selections restricted to the highest ranked sites. The mix of information IMPROvER uses can be extracted for any helical membrane protein. We have developed the first general-purpose tool for selecting stabilising variants of [Formula: see text]-helical membrane proteins, increasing efficiency and reducing workload. IMPROvER can be accessed at http://improver.ddns.net/IMPROvER/ .

Late and early leaf spot are caused by Nothopassalora personata and Passalora arachidicola , respectively, and are damaging diseases of peanut ( Arachis hypogaea L.) capable of defoliation and yield loss. Management of these diseases is most effective through the integration of tactics that reduce starting inoculum and prevent infection. The insecticide phorate was first registered in 1959 and has been used in peanut production for decades in-furrow at planting to suppress thrips. Phorate further provides significant suppression of Tomato spotted wilt virus infection beyond suppression of its thrips vector alone by activating defense-related responses in the peanut plant. From six experiments conducted from 2017 to 2019 in Blackville, SC, Reddick, FL, and Quincy, FL, significantly less leaf spot defoliation was exhibited on peanuts treated with phorate in-furrow at planting (26%) compared with nontreated checks (48%). In-season fungicides were excluded from five of the experiments, whereas the 2018 Quincy, FL, experiment included eight applications on a 15-day interval. Across individual experiments, significant suppression of defoliation caused by late leaf spot was observed from 64 to 147 days after planting. Although more variable within location-years, pod yield following phorate treatment was overall significantly greater than for nontreated peanut (2,330 compared with 2,030 kg/ha; P = 0.0794). The consistent defoliation suppression potential was estimated to confer an average potential net economic yield savings of $90 to $120 per hectare under analogous leaf spot defoliation. To our knowledge, these are the first data in the 61 years since its registration demonstrating significant suppression of leaf spot on peanut following application of phorate in-furrow at planting. Results support phorate use in peanut as an effective and economical tactic to incorporate to manage late and early leaf spot infections and development of fungicide resistance.

The neural systems that govern declarative and procedural memory processing do not always operate independently. Direct evidence of competition between these two memory systems in humans is supported by studies showing that performing a declarative learning task immediately after motor skill learning can disrupt procedural memory and abolish the off-line gains in skill performance obtained during consolidation. The aim of the present study was to extend recent investigations demonstrating that the exposure to a brief bout of cardiovascular exercise can protect procedural memory by enhancing postpractice consolidation. We used an experimental paradigm designed to assess whether exercise can also protect procedural memory consolidation from interference induced with declarative learning. The implicit acquisition of a serial reaction time task (SRTT) was tested after a 6-h waked-filled period. Participants who were exposed to a non-learning vowel counting (VC) task following the practice of the SRTT exhibited successful procedural memory consolidation and significant off-line gains in skill performance. Confirming that declarative memory processes can interfere with procedural memory consolidation, off-line gains in motor skill performance were suppressed when the performance of the VC task was replaced with a word list (WL) task requiring declarative learning. Performing a bout of cardiovascular exercise after the SRTT protected the newly formed procedural memory from the interference produced by the WL task. Protection was evidenced by a return of significant off-line gains in skill performance after the waked-filled period. Exercise optimizes the utilization of neural resources reducing interference between procedural and declarative memory systems., (Copyright © 2020 Chen et al.)

Interleaved rather than repetitive practice (RP) is associated with superior retention of motor skills. It has been argued that this results from improved post-practice consolidation reflected in greater offline gains following interleaved practice (IP). The magnitude of this offline benefit has been associated with greater recruitment of supplementary motor area (SMA) during encoding. The present study administered anodal or cathodal transcranial direct current stimulation (tDCS) during interleaved or RP respectively in an attempt to modify the activity at SMA and the concomitant retention outcomes commonly associated with these training formats. Sixty-nine participants were assigned to one of four experimental conditions that included: IP-sham, RP-sham, IP-cathodal tDCS, and RP-anodal tDCS. Real or sham stimulation at SMA was administered during practice of three unique 6-key discrete sequence production tasks which lasted approximately 20-min. Tests were administered prior to practice and immediately after practice as well at 6-h, 24-h, and 72-h after practice ended. As anticipated, IP resulted in poorer acquisition but superior offline gain. Enhanced offline gain following interleaved training resulted from rapid stabilization of performance within the first 6-h following encoding and overnight improvement that continued over multiple sleep episodes. Administration of anodal stimulation at SMA during RP improved performance during training compared to sham but this benefit was short lived as forgetting during the first 6-h after practice was consistent with that observed for the sham counterpart. However, supplementing RP with anodal stimulation at SMA did foster overnight offline performance gains not displayed by individuals that experienced RP in the absence of stimulation., (Copyright © 2020 IBRO. Published by Elsevier Ltd. All rights reserved.)

The purpose of the study was twofold: (1) determine if different time delays (30 min or 6 h) between training and a post-training test with a rhythmic bimanual pattern (90° relative phase) would be associated with different levels of consolidation for the motor and spatial representations of the pattern; and (2) determine if training with the rhythmic bimanual pattern would lead to enhanced corticospinal excitability in M1 linked to changes in motor and spatial performance measures. Coordination accuracy and stability of the 90° pattern improved over practice. Coordination accuracy and stability were the same after a 30-min or 6-h delay between training and the post-training test, indicating equivalent levels of consolidation in the motor representation. The 6-h delay interval resulted in shorter visual recognition times compared to the 30-min delay and was centered on the trained 90° pattern. These findings indicate the consolidation of the spatial representation was more time sensitive compared to the motor representation in the current task. Motor evoked potentials (MEPs) from the first dorsal interosseous muscle (FDI) generated by single-pulse transcranial magnetic stimulation (TMS) were measured at baseline (before training) and at 6-min and 21-min intervals post-training with the 90° pattern. Increased corticospinal excitability in M1 was evidenced by larger MEPs of the FDI muscle at the 6-min interval. This increased excitability after training is a novel finding after training with a difficult and initially unstable rhythmic bimanual pattern. No significant correlations were found between the MEP data and behavioral data; thus, the increased excitability in M1 may have been linked to the difficulty in performing the pattern, consolidation processes, or both.

Late and early leaf spot, respectively caused by Nothopassalora personata and Passalora arachidicola , are damaging diseases of peanut ( Arachis hypogaea ) capable of defoliating canopies and reducing yield. Although one of these diseases may be more predominant in a given area, both are important on a global scale. To assist informed management decisions and quantify relationships between end-of-season defoliation and yield loss, meta-analyses were conducted over 140 datasets meeting established criteria. Slopes of proportion yield loss with increasing defoliation were estimated separately for Virginia and runner market type cultivars. Yield loss for Virginia types was described by an exponential function over the range of defoliation levels, with a loss increase of 1.2 to 2.2% relative to current loss levels per additional percent defoliation. Results for runner market type cultivars showed yield loss to linearly increase 2.2 to 2.8% per 10% increase in defoliation for levels up to approximately 95% defoliation, after which the rate of yield loss was exponential. Defoliation thresholds to prevent economic yield loss for Virginia and runner types were estimated at 40 and 50%, respectively. Although numerous factors remain important in mitigating overall yield losses, the integration of these findings should aid recommendations about digging under varying defoliation intensities and peanut maturities to assist in minimizing yield losses.

Two experiments examined whether practicing discrete key pressing sequences eventually leads to a disregard of the key-specific stimuli, as suggested by sequence learning models, or whether these stimuli continue to be relied upon because the associated luminance increase attracts visuospatial attention. Participants practiced two sequences by reacting to two fixed series of seven letter stimuli, each displayed at a location that did or did not correspond with the required response location. Stimulus use was indicated by a Simon effect in that key presses were slowed when stimulus and key locations did not correspond. Experiment 1 demonstrated that letter stimuli continued to be used as the Simon effect occurred with each sequence element, and this remained quite stable across practice and did not differ for familiar and unfamiliar sequences. Experiment 2 showed that the Simon effect remained present even with meaningless stimuli that were often even harmful. These findings suggest that even in motor sequences that can be executed without element-specific stimuli attention attraction enforces stimulus use. The data further supported the assumptions that S-R translation and sequencing systems are racing to trigger individual responses, and that explicit sequence representations include spatial and verbal knowledge., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

Plant-parasitic nematodes ( Rotylenchulus reniformis (reniform, RN), Helicotylenchus dihystera (spiral), and Mesocriconema ornatum (ring)) and yield were investigated in cotton phases of conventional (peanut-cotton-cotton) and sod-based (bahiagrass-bahiagrass-peanut-cotton) rotations with or without irrigation and fluopyram nematicide at a long-term research site, established in 2000, in Quincy, Florida, USA. Objectives were to determine impacts of nematicide application on cotton yield and evaluate effects of nematicide on plant-parasitic nematodes in these rotations in 2017 and 2018. Reniform nematode population densities were greater in conventional cotton than sod-based cotton. Ring and spiral nematode population densities were greater in sod-based cotton than conventional cotton. Plots receiving nematicide had increased RN population densities in preplant 2018 soil samples and spiral nematode population densities in preplant 2017, harvest 2017, preplant 2018, and harvest 2018 soil samples compared to untreated plots. Cotton seed yield in conventional rotation was increased by 18% following nematicide application in 2017 but decreased by 10% in sod-based rotation in 2018, relative to the untreated control. Sod-based rotation had greater cotton yield than conventional rotation in 2017 and 2018. Nematicide application did not improve cotton yield in sod-based rotation and was inconsistent in conventional rotation., Plant-parasitic nematodes ( Rotylenchulus reniformis (reniform, RN), Helicotylenchus dihystera (spiral), and Mesocriconema ornatum (ring)) and yield were investigated in cotton phases of conventional (peanut–cotton–cotton) and sod-based (bahiagrass–bahiagrass–peanut–cotton) rotations with or without irrigation and fluopyram nematicide at a long-term research site, established in 2000, in Quincy, Florida, USA. Objectives were to determine impacts of nematicide application on cotton yield and evaluate effects of nematicide on plant-parasitic nematodes in these rotations in 2017 and 2018. Reniform nematode population densities were greater in conventional cotton than sod-based cotton. Ring and spiral nematode population densities were greater in sod-based cotton than conventional cotton. Plots receiving nematicide had increased RN population densities in preplant 2018 soil samples and spiral nematode population densities in preplant 2017, harvest 2017, preplant 2018, and harvest 2018 soil samples compared to untreated plots. Cotton seed yield in conventional rotation was increased by 18% following nematicide application in 2017 but decreased by 10% in sod-based rotation in 2018, relative to the untreated control. Sod-based rotation had greater cotton yield than conventional rotation in 2017 and 2018. Nematicide application did not improve cotton yield in sod-based rotation and was inconsistent in conventional rotation.

Tecchio et al. (J Neurophysiology 104: 1134-1140, 2010) reported that the application of anodal tDCS at primary motor cortex (M1) immediately after practice of a procedural motor skill enhanced consolidation, which in turn improved offline gain. Tecchio et al. noted, however, that this study did not account for known after-effects associated with this form of non-invasive stimulation. The present study was designed to explicitly reevaluate Tecchio et al.'s claim. As in the original study, individuals experienced either anodal or sham stimulation at M1 after practice of a serial reaction time task (SRTT) followed by test trials 15-min later. Two additional novel conditions experienced the test trials after 120-min rather than 15-min thus allowing potential stimulation after-effects to dissipate. The expectation was that if anodal stimulation influences post-practice consolidation leading to offline gain, this effect would be present not only at 15-min but also after 120-min. In agreement with the working hypothesis, findings revealed offline gain at both 15-min and the longer 2-h time period. Unexpectedly, we found no interaction between real and sham conditions. The lack of difference between Real and Sham effects weakens confidence in the potential of post-practice tDCS for consolidation enhancement, while it is more consistent with other claims that decoupling practice and anodal tDCS stimulation in time can reduce the effectiveness of exogenous stimulation for procedural skill gain.

Many years ago, the natural secondary metabolite SF2312, produced by the actinomycete Micromonospora, was reported to display broad spectrum antibacterial properties against both Gram-positive and Gram-negative bacteria. Recent studies have revealed that SF2312, a natural phosphonic acid, functions as a potent inhibitor of human enolase. The mechanism of SF2312 inhibition of bacterial enolase and its role in bacterial growth and reproduction, however, have remained elusive. In this work, we detail a structural analysis of E. coli enolase bound to both SF2312 and its oxidized imide-form. Our studies support a model in which SF2312 acts as an analog of a high energy intermediate formed during the catalytic process. Biochemical, biophysical, computational and kinetic characterization of these compounds confirm that altering features characteristic of a putative carbanion (enolate) intermediate significantly reduces the potency of enzyme inhibition. When SF2312 is combined with fosfomycin in the presence of glucose-6 phosphate, significant synergy is observed. This suggests the two agents could be used as a potent combination, targeting distinct cellular mechanism for the treatment of bacterial infections. Together, our studies rationalize the structure-activity relationships for these phosphonates and validate enolase as a promising target for antibiotic discovery.

The spread of plasmid borne resistance enzymes in clinical Staphylococcus aureus isolates is rendering trimethoprim and iclaprim, both inhibitors of dihydrofolate reductase (DHFR), ineffective. Continued exploitation of these targets will require compounds that can broadly inhibit these resistance-conferring isoforms. Using a structure-based approach, we have developed a novel class of ionized nonclassical antifolates (INCAs) that capture the molecular interactions that have been exclusive to classical antifolates. These modifications allow for a greatly expanded spectrum of activity across these pathogenic DHFR isoforms, while maintaining the ability to penetrate the bacterial cell wall. Using biochemical, structural, and computational methods, we are able to optimize these inhibitors to the conserved active sites of the endogenous and trimethoprim resistant DHFR enzymes. Here, we report a series of INCA compounds that exhibit low nanomolar enzymatic activity and potent cellular activity with human selectivity against a panel of clinically relevant TMP resistant (TMP R ) and methicillin resistant Staphylococcus aureus (MRSA) isolates.

Numerous studies have reported a positive impact of acute exercise for procedural skill memory. Previous work has revealed this effect, but these findings are confounded by a potential contribution of a night of sleep to the reported exercise-mediated reduction in interference. Thus, it remains unclear if exposure to a brief bout of exercise can provide protection to a newly acquired motor memory. The primary objective of the present study was to examine if a single bout of moderate-intensity cardiovascular exercise after practice of a novel motor sequence reduces the susceptibility to retroactive interference. To address this shortcoming, 17 individuals in a control condition practiced a novel motor sequence that was followed by test after a 6-h wake-filled interval. A separate group of 17 individuals experienced practice with an interfering motor sequence 45 min after practice with the original sequence and were then administered test trials 6 h later. One additional group of 12 participants was exposed to an acute bout of exercise immediately after practice with the original motor sequence but prior to practice with the interfering motor sequence and the subsequent test. In comparison with the control condition, increased response times were revealed during the 6-h test for the individuals that were exposed to interference. The introduction of an acute bout of exercise between the practice of the two motor sequences produced a reduction in interference from practice with the second task at the time of test, however, this effect was not statistically significant. These data reinforce the hypothesis that while there may be a contribution from exercise to post-practice consolidation of procedural skills which is independent of sleep, sleep may interact with exercise to strengthen the effects of the latter on procedural memory.

Pharmacokinetic/pharmacodynamic properties are strongly correlated with the in vivo efficacy of antibiotics. Propargyl-linked antifolates, a novel class of antibiotics, demonstrate potent antibacterial activity against both Gram-positive and Gram-negative pathogenic bacteria, including multidrug-resistant Staphylococcus aureus Here, we report our efforts to optimize the pharmacokinetic profile of this class to best match the established pharmacodynamic properties. High-resolution crystal structures were used in combination with in vitro pharmacokinetic models to design compounds that not only are metabolically stable in vivo but also retain potent antibacterial activity. The initial lead compound was prone to both N -oxidation and demethylation, which resulted in an abbreviated in vivo half-life (∼20 minutes) in mice. Stability of leads toward mouse liver microsomes was primarily used to guide medicinal chemistry efforts so robust efficacy could be demonstrated in a mouse disease model. Structure-based drug design guided mitigation of N -oxide formation through substitutions of sterically demanding groups adjacent to the pyridyl nitrogen. Additionally, deuterium and fluorine substitutions were evaluated for their effect on the rate of oxidative demethylation. The resulting compound was characterized and demonstrated to have a low projected clearance in humans with limited potential for drug-drug interactions as predicted by cytochrome P450 inhibition as well as an in vivo exposure profile that optimizes the potential for bactericidal activity, highlighting how structural data, merged with substitutions to introduce metabolic stability, are a powerful approach to drug design., (Copyright © 2019 by The Author(s).)

This paper describes song production by the eastern North Pacific right whale (NPRW, Eubalaena japonica) in the southeastern Bering Sea. Songs were localized in real-time to individuals using sonobuoys. Singers whose sex could be determined were all males. Autonomous recorder data from 17 year-long deployments were analyzed to document and characterize song types. Four distinct song types were documented over eight years (2009-2017) at five distinct locations. Each song type consists of a hierarchical structure of 1-3 different repeating phrases comprised predominantly of gunshot sounds; three of the four songs contained additional sound types (downsweep, moan, and low-frequency pulsive call). Songs were detected annually (July-January); all song types remained consistent over eight years. Two different songs often occurred simultaneously, produced by different individuals; the same song was never detected simultaneously at the same location. The same song type was detected on the same day and time at two distant locations, indicating multiple individuals can produce the same song. These findings provide support that males produce song; it remains unknown if females also sing. NPRW is the first right whale species documented to produce song. Based on current knowledge about song in mysticetes, it is hypothesized that these songs are reproductive displays.

Enolase is a glycolytic metalloenzyme involved in carbon metabolism. The advantage of targeting enolase lies in its essentiality in many biological processes such as cell wall formation and RNA turnover and as a plasminogen receptor. We initially used a DARTS assay to identify enolase as a target in Escherichia coli. The antibacterial activities of α-, β-, and γ-substituted seven-member ring tropolones were first evaluated against four strains representing a range of Gram-negative bacteria. We observed that the chemical properties and position of the substituents on the tropolone ring play an important role in the biological activity of the investigated compounds. Both α- and β-substituted phenyl derivatives of tropolone were the most active with minimum inhibitory concentrations in the range of 11-14 μg/mL. The potential inhibitory activity of the synthetic tropolones was further evaluated using an enolase inhibition assay, X-ray crystallography, and molecular docking simulations. The catalytic activity of enolase was effectively inhibited by both the naturally occurring β-thujaplicin and the α- and β-substituted phenyl derivatives of tropolones with IC 50 values in range of 8-11 μM. Ligand binding parameters were assessed by isothermal titration calorimetry and differential scanning calorimetry techniques and agreed with the in vitro data. Our studies validate the antibacterial potential of tropolones with careful consideration of the position and character of chelating moieties for stronger interaction with metal ions and residues in the enolase active site.

Tropolones are naturally occurring seven-membered non-benzenoid aromatic compounds that are of interest due to their cytotoxic properties. MO-OH-Nap is a novel α-substituted tropolone that induces caspase cleavage and upregulates markers associated with the unfolded protein response (UPR) in multiple myeloma (MM) cells. Given previous reports that tropolones may function as iron chelators, we investigated the effects of MO-OH-Nap, as well as the known iron chelator deferoxamine (DFO), in MM cells in the presence or absence of supplemental iron. The ability of MO-OH-Nap to induce apoptosis and upregulate markers of the UPR could be completely prevented by co-incubation with either ferric chloride or ammonium ferrous sulfate. Iron also completely prevented the decrease in BrdU incorporation induced by either DFO or MO-OH-Nap. Ferrozine assays demonstrated that MO-OH-Nap directly chelates iron. Furthermore, MO-OH-Nap upregulates cell surface expression and mRNA levels of transferrin receptor. In vivo studies demonstrate increased Prussian blue staining in hepatosplenic macrophages in MO-OH-Nap-treated mice. These studies demonstrate that MO-OH-Nap-induced cytotoxic effects in MM cells are dependent on the tropolone's ability to alter cellular iron availability and establish new connections between iron homeostasis and the UPR in MM., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

AK3 compounds are mitotic arrest agents that induce high levels of γH2AX during mitosis and apoptosis following release from arrest. We synthesized a potent AK3 derivative, AK306, that induced arrest and apoptosis of the HCT116 colon cancer cell line with an EC 50 of approximately 50 nmol/L. AK306 was active on a broad spectrum of cancer cell lines with total growth inhibition values ranging from approximately 25 nmol/L to 25 μmol/L. Using biotin and BODIPY-linked derivatives of AK306, binding to clathrin heavy chain (CLTC/CHC) was observed, a protein with roles in endocytosis and mitosis. AK306 inhibited mitosis and endocytosis, while disrupting CHC cellular localization. Cells arrested in mitosis by AK306 showed the formation of multiple microtubule-organizing centers consisting of pericentrin, γ-tubulin, and Aurora A foci, without apparent centrosome amplification. Cells released from AK306 arrest were unable to form bipolar spindles, unlike nocodazole-released cells that reformed spindles and completed division. Like AK306, CHC siRNA knockdown disrupted spindle formation and activated p53. A short-term (3-day) treatment of tumor-bearing APC -mutant mice with AK306 increased apoptosis in tumors, but not normal mucosa. These findings indicate that targeting the mitotic CHC complex can selectively induce apoptosis and may have therapeutic value. Implication: Disruption of clathrin with a small-molecule inhibitor, AK306, selectively induces apoptosis in cancer cells by disrupting bipolar spindle formation. Mol Cancer Res; 16(9); 1361-72. ©2018 AACR ., (©2018 American Association for Cancer Research.)

The influence of monitoring-pressure and verbal feedback on the performance of the intrinsically stable bimanual coordination patterns of in-phase and anti-phase was examined. The two bimanual patterns were produced under three conditions: 1) no-monitoring, 2) monitoring-pressure (viewed by experts), and 3) monitoring-pressure (viewed by experts) combined with verbal feedback emphasizing poor performance. The bimanual patterns were produced at self-paced movement frequencies. Anti-phase coordination was always less stable than in-phase coordination across all three conditions. When performed under conditions 2 and 3, both bimanual patterns were performed with less variability in relative phase across a wide range of self-paced movement frequencies compared to the no-monitoring condition. Thus, monitoring-pressure resulted in performance stabilization rather than degradation and the presence of verbal feedback had no impact on the influence of monitoring pressure. The current findings are inconsistent with the predictions of explicit monitoring theory; however, the findings are consistent with studies that have revealed increased stability for the system's intrinsic dynamics as a result of attentional focus and intentional control. The results are discussed within the contexts of the dynamic pattern theory of coordination, explicit monitoring theory, and action-focused theories as explanations for choking under pressure., (Copyright © 2018. Published by Elsevier B.V.)

Objective: To identify the impact of embryo transfer time (total seconds from the loading of the transfer catheter to the expulsion of the embryo(s) into the uterine cavity) on clinical pregnancy (CPR), implantation (IR), and live birth (LBR) rates., Design: Retrospective cohort study., Setting: Academic hospital practice., Patient(s): A total of 465 women undergoing 571 frozen-embryo transfers with the use of cryopreserved blastocysts in a single academic institution from 2007 through 2014., Intervention(s): None., Main Outcome Measure(s): CPR, IR, and LBR., Result(s): The cohort was divided into tertiles according to transfer time in seconds (T1: 33-55; T2: 57-81; T3: 82-582) with mean (SD) transfer times of 47.4 (5.7), 67.1 (7.3), and 121.9 (55.1) seconds, respectively. Crude CPRs were 43.9%, 48.7%, and 48.7% among the respective tertiles, crude IRs were 36.9%, 39.9%, and 38.6%, and crude LBRs were 34.8%, 39.6%, and 36.0%. In univariate analysis, inferior cohort score, blood inside catheter, difficult mock transfer, and use of an outer sheath were negatively associated with CPR. No association was seen between physician performing the transfer (including fellows) and CPR. In multivariate regression, longer transfer time was not associated with CPR. With T1 as reference, adjusted odds ratios (95% confidence interval) were 1.28 (0.77-2.11) and 1.52 (0.85-2.71) for transfer time groups T2 and T3, respectively., Conclusion(s): After adjusting for potential confounders, this analysis found that contrary to commonly held belief, longer embryo transfer times do not negatively affect CPR, IR, or LBR., (Copyright © 2017 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.)

The primary objective of the present experiment was to examine the influence of recent practice in a random and blocked format for future motor learning. First, individuals practiced three unique discrete sequence production tasks in either a blocked or random schedule. One day later, all individuals practiced a new motor sequence not previously practiced. On day three, mean total time for the test performance of the original three motor sequences was lower for individuals that practiced in a random format. This emerged as a significant reduction in mean total time from the completion of practice and the test trials implicating offline consolidation as a key contributor to the random practice performance advantage. A novel finding from the present work was that the acquisition of the novel discrete sequence production task practiced on Day 2 was better for individuals that had prior random rather than blocked practice experience. This benefit was robust appearing early during acquisition as significantly lower mean total time. This benefit from random practice experience remained during the delayed test trials administered on Day 3 for the novel motor sequence., (Published by Elsevier B.V.)

The herpes simplex virus (HSV) type I alkaline nuclease, UL12, has 5'-to-3' exonuclease activity and shares homology with nucleases from other members of the Herpesviridae family. We previously reported that a UL12-null virus exhibits a severe defect in viral growth. To determine whether the growth defect was a result of loss of nuclease activity or another function of UL12, we introduced an exonuclease-inactivating mutation into the viral genome. The recombinant virus, UL12 D340E (the D340E mutant), behaved identically to the null virus (AN-1) in virus yield experiments, exhibiting a 4-log decrease in the production of infectious virus. Furthermore, both viruses were severely defective in cell-to-cell spread and produced fewer DNA-containing capsids and more empty capsids than wild-type virus. In addition, DNA packaged by the viral mutants was aberrant, as determined by infectivity assays and pulsed-field gel electrophoresis. We conclude that UL12 exonuclease activity is essential for the production of viral DNA that can be packaged to produce infectious virus. This conclusion was bolstered by experiments showing that a series of natural and synthetic α-hydroxytropolones recently reported to inhibit HSV replication also inhibit the nuclease activity of UL12. Taken together, our results demonstrate that the exonuclease activity of UL12 is essential for the production of infectious virus and may be considered a target for development of antiviral agents. IMPORTANCE Herpes simplex virus is a major pathogen, and although nucleoside analogs such as acyclovir are highly effective in controlling HSV-1 or -2 infections in immunocompetent individuals, their use in immunocompromised patients is complicated by the development of resistance. Identification of additional proteins essential for viral replication is necessary to develop improved therapies. In this communication, we confirm that the exonuclease activity of UL12 is essential for viral replication through the analysis of a nuclease-deficient viral mutant. We demonstrate that the exonuclease activity of UL12 is essential for the production of viral progeny and thus provides an attractive, druggable enzymatic target., (Copyright © 2017 American Society for Microbiology.)

Tropolones are small organic compounds with metal-directing moieties. Tropolones inhibit the proliferation of cancer cell lines, possibly through their effects on metalloenzymes such as select histone deacetylases (HDACs). Pan-HDAC inhibitors are therapeutically beneficial in the treatment of multiple myeloma, however there is interest in the use of more selective HDAC inhibitor therapy to minimize adverse side effects. We hypothesized that tropolones might have anti-myeloma activities. To this end, a series of novel α-substituted tropolones were evaluated for effects on multiple myeloma cells. While all tested tropolones showed some level of cytotoxicity, MO-OH-Nap had consistently low IC 50 values between 1-11 μM in all three cell lines tested and was used for subsequent experiments. MO-OH-Nap was found to induce apoptosis in a concentration-dependent manner. Time course experiments demonstrated that MO-OH-Nap promotes caspase cleavage in a time frame that was distinct from the pan-HDAC inhibitor suberoylanilide hydroxamic acid (SAHA). Furthermore, MO-OH-Nap- and SAHA-treated cells possess unique gene expression patterns, suggesting they promote apoptosis via different mechanisms. In particular, MO-OH-Nap increases the expression of markers associated with endoplasmic reticulum stress and the unfolded protein response. Synergistic cytotoxic effects were observed when cells were treated with the combination of MO-OH-Nap and the proteasome inhibitor bortezomib. However, treatment with MO-OH-Nap did not abrogate the bortezomib-induced increase in aggresomes, consistent with an HDAC6-independent mechanism for the observed synergy. Collectively, these finding support further investigation into the usefulness of α-substituted tropolones as anti-myeloma agents., Competing Interests: CONFLICTS OF INTEREST The authors have no relevant conflicts of interest to disclose.

The influence of monitoring-pressure on the performance of anti-phase and in-phase bimanual coordination was examined. The two bimanual patterns were produced under no-monitoring and monitoring-pressure conditions at self-paced frequencies. Anti-phase coordination was always less stable than in-phase coordination, with or without monitoring. When performed under monitoring-pressure, the coordination patterns were performed with less variability in relative phase for both patterns across a range of self-paced movement frequencies compared to performance without monitoring. Thus, while monitoring-pressure did induce a behavioral change, it consisted of performance stabilization rather than degradation, a finding inconsistent with explicit-monitoring theory. However, the findings are consistent with the theory of coordination dynamics and studies that have revealed increased stability for the system's intrinsic dynamics as a result of attentional focus and intentional control.

Antimicrobial resistance to current antibiotics is a significant public health problem and the need for new antibiotics is a compelling one. We have been developing a new series of antibiotics, propargyl-linked diaminopyrimidines, based on the structure of trimethoprim. To date we have discovered compounds that are effective inhibitors of dihydrofolate reductase (the target of trimethoprim), that are potent antibiotics in vitro against a range of Gram-positive pathogens including methicillin-resistant S. aureus, and that are non-toxic in mammalian cell culture. In this study we report the development of an LC-MS-based protocol for the quantification of our lead antibiotic 37D1-UCP1099 and the application of this assay to follow the concentration of the compound in mouse plasma after intraperitoneal administration. Extraction of 37D1-UCP1099 from mouse plasma was achieved through a liquid-liquid extraction with ethyl acetate. Separation was performed utilizing a reverse-phase C18 column with a ten minute isocratic elution using 47:53 (v/v) 10mM NH 4 HCO 3 :acetonitrile. The lower limit of quantitation for 37D1-UCP1099 was 50ngmL -1 and the assay showed a dynamic range of 50-4000ngmL -1 with good linearity (r 2 ≥0.996 for all fits). Intra-day and inter-day precision and accuracy were within 11.3% (%RSD) and 6.6% (%RE) respectably. We have demonstrated that the compound is stable under the assay procedures. The compound was shown to have a mean residence time of 26.2±1.0min and a half-life of 18.2±0.7min after intraperitoneal delivery at 5mgkg -1 . These studies now form the foundation of our work to develop additional analogs of 37D1-UCP1099 with improved pharmacokinetic properties., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Much of the literature on the impact of male caffeine and alcohol intake on reproductive outcomes has utilized semen quality as a proxy for male fertility, although semen parameters have a limited predictive value for spontaneous pregnancy. The objective of this study was to investigate whether male caffeine and alcohol intakes are associated with semen parameters and assisted reproductive technology outcome. The Environment and Reproductive Health Study, an ongoing prospective cohort study, enrolls subfertile couples presenting for treatment at an academic fertility center (2007-2012). A total of 171 men with 338 semen analyses and 205 assisted reproductive technology cycles were included in this analysis. Diet was assessed using a 131-item food frequency questionnaire. Mixed models adjusting for potential confounders were used to evaluate the relationships of male caffeine and alcohol intakes with semen parameters and assisted reproductive technology outcomes. There was no association between male caffeine and alcohol intake and semen quality. Male caffeine intake was negatively related to live birth after assisted reproductive technologies (p-trend < 0.01), and male alcohol intake was positively related to live birth after assisted reproductive technologies (p-trend = 0.04). Adjusted live birth rate among couples with a male partner in the highest quartile of caffeine intake (≥272 mg/day) compared to couples with a male partner in the lowest quartile of intake (<99 mg/day) was 19% vs. 55%, respectively, p < 0.01. In terms of alcohol intake, adjusted live birth rate among couples with a male partner in the highest quartile of alcohol intake (≥22 g/day) compared to couples with a male partner in the lowest quartile of intake (<3 g/day) was 61% vs. 28%, respectively, p = 0.05. In conclusion, male pre-treatment caffeine and alcohol intakes were associated with live birth after assisted reproductive technologies, but not with semen parameters, among fertility patients., (© 2017 American Society of Andrology and European Academy of Andrology.)

To develop next generation antifolates for the treatment of trimethoprim-resistant bacteria, synthetic methods were needed to prepare a diverse array of 3-aryl-propynes with various substitutions at the propargyl position. A direct route was sought whereby nucleophilic addition of acetylene to aryl carboxaldehydes would be followed by reduction or substitution of the resulting propargyl alcohol. The direct reduction, methylation, and dimethylation of these readily available alcohols provide efficient access to this uncommon functional array. In addition, an unusual silane exchange reaction was observed in the reduction of the propargylic alcohols.

Antibiotic resistance is a rapidly evolving health concern that requires a sustained effort to understand mechanisms of resistance and to develop new agents that overcome those mechanisms. The dihydrofolate reductase (DHFR) inhibitor, trimethoprim (TMP), remains one of the most important orally administered antibiotics. However, resistance through chromosomal mutations and mobile, plasmid-encoded insensitive DHFRs threatens the continued use of this agent. We are pursuing the development of new propargyl-linked antifolate (PLA) DHFR inhibitors designed to evade these mechanisms. While analyzing contemporary TMP-resistant clinical isolates of methicillin-resistant and sensitive Staphylococcus aureus, we discovered two mobile resistance elements, dfrG and dfrK. This is the first identification of these resistance mechanisms in the United States. These resistant organisms were isolated from a variety of infection sites, show clonal diversity, and each contain distinct resistance genotypes for common antibiotics. Several PLAs showed significant activity against these resistant strains by direct inhibition of the TMP resistance elements., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Tropolones, such as β-thujaplicin, are small lead-like natural products that possess a variety of biological activities. While the β-substituted natural products and their synthetic analogs are potent inhibitors of human cancer cell growth, less is known about their α-substituted counterparts. Recently, we synthesized a series of α-substituted tropolones including 2-hydroxy-7-(naphthalen-2-yl)cyclohepta-2,4,6-trien-1-one (α-naphthyl tropolone). Here, we evaluate the antiproliferative mechanisms of α-naphthyl tropolone and the related α-benzodioxinyl analog. The α-substituted tropolones inhibit growth of lymphocytic leukemia cells, but not healthy blood cells, with nanomolar potency. Treatment of leukemia cell lines with the tropolone dose-dependently induces apoptosis as judged by staining with annexin V and propidium iodide and Western blot analysis of cleaved caspase 3 and 7. Moreover, pre-treatment of cells with the caspase inhibitor Z-VAD-FMK inhibited the apoptotic effects of the tropolone in two lymphocytic lines. Caspase inhibition also blocked elevated histone acetylation caused by the tropolone, indicating that its effects on histone acetylation are potentiated by caspases. In contrast, α-naphthyl tropolone upregulated p53 expression and phosphorylation of Akt and mTOR in a manner that was not rescued by caspase inhibition. The effects of tropolone were blocked by co-incubation with high levels of free extracellular iron but not by pre-loading with iron. Additionally, dose and time dependent reduction in ex vivo viability of cells from leukemia patients was observed. Taken together, we demonstrate that α-substituted tropolones upregulate DNA damage repair pathways leading to caspase-dependent apoptosis in malignant lymphocytes., Competing Interests: statement A.J.W. is a co-founder of Terpenoid Therapeutics. The current work did not involve the company., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

In healthy young adults motor learning is typically improved by practice under conditions of high contextual interference such as that created when a series of skills are practiced in a random sequence. The authors examined whether individuals with Parkinson's disease and age-matched older adults might also benefit from such a learning environment. Participants learned 3 peg placement movements under both random practice and blocked practice schedules. Participants completed free-recall, cued-recall, and transfer tests 1 day and 1 week following practice. During acquisition all learners had shorter movement times (MTs) during blocked than random practice but during retention testing, MTs were shorter if the patterns had been learned with random practice. Fewer errors during free recall following random practice were evident in both groups of learners and MTs to complete a novel transfer task were shorter following random practice. These findings suggest employing a cognitively challenging practice environment might improve motor skill learning in elderly learners and in people with Parkinson's disease.

Cryopreservation of human spermatozoa is a commonly used technique in assisted reproduction, however freezing low concentrations of sperm while maintaining adequate post-thaw motility remains a challenge. In an effort to optimize post-thaw motility yields, low volumes of human sperm were frozen in polyimide-coated fused silica micro-capillaries using 0.065 M, 0.125 M, 0.25 M, or 0.5 M trehalose as the only cryoprotectant. Micro-capillaries were either initially incubated in liquid nitrogen vapor before plunging into liquid nitrogen, or directly plunged into liquid nitrogen. Post thaw sperm counts and motility were estimated. Spermatozoa that were initially incubated in liquid nitrogen vapor had greater post thaw motility than those plunged immediately into liquid nitrogen independent of trehalose concentration. The protective effect of 0.125 M d-glucose, 3-O-methyl-d-glucopyranose, trehalose, sucrose, raffinose, or stachyose were evaluated individually. Trehalose and sucrose were the most effective cryoprotectants, recovering 69.0% and 68.9% of initial sperm motility, respectively., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Background: Vitamin D deficiency impairs fertility in animal models, but the role of vitamin D in human fertility or treatment of infertility is less clear., Objective: We examined the association between circulating 25-hydroxyvitamin D [25(OH)D] concentrations and the outcome in women undergoing assisted reproduction technologies (ARTs)., Design: We randomly selected 100 women undergoing infertility treatment with ART enrolled in an ongoing prospective cohort study who underwent 168 treatment cycles. Serum 25(OH)D concentrations were measured in samples collected from women between days 3 and 9 of gonadotropin treatment. Generalized linear mixed models were used to evaluate the association of 25(OH)D concentrations with ART outcomes while adjusting for potential confounders and accounting for repeated treatment cycles per woman., Results: Median (range) serum 25(OH)D concentrations were 86.5 (33.5-155.5) nmol/L. Ninety-one percent of participants consumed multivitamins. Serum 25(OH)D concentrations were positively related to fertilization rate. The adjusted fertilization rate for women in increasing quartiles of serum 25(OH)D were 0.62 (95% CI: 0.51, 0.72), 0.53 (95% CI: 0.43, 0.63), 0.67 (95% CI: 0.56, 0.76), and 0.73 (95% CI: 0.63, 0.80), respectively (P-trend = 0.03). This association persisted when analyses were restricted to women with serum 25(OH)D between 50 and 125 nmol/L when models were further adjusted for season of blood draw and when analyses were restricted to the first treatment cycle. However, 25(OH)D concentrations were unrelated to probability of pregnancy (P-trend = 0.83) or live birth after ART (P-trend = 0.47)., Conclusion: Vitamin D may be associated with higher fertilization rates, but this apparent benefit does not translate into higher probability of pregnancy or live birth. This trial was registered at www.clinicaltrials.gov as NCT00011713., (© 2016 American Society for Nutrition.)

Mycobacterium tuberculosis continues to cause widespread, life-threatening disease. In the last decade, this threat has grown dramatically as multi- and extensively-drug resistant (MDR and XDR) bacteria have spread globally and the number of agents that effectively treat these infections is significantly reduced. We have been developing the propargyl-linked antifolates (PLAs) as potent inhibitors of the essential enzyme dihydrofolate reductase (DHFR) from bacteria and recently found that charged PLAs with partial zwitterionic character showed improved mycobacterial cell permeability. Building on a hypothesis that these PLAs may penetrate the outer membrane of M. tuberculosis and inhibit the essential cytoplasmic DHFR, we screened a group of PLAs for antitubercular activity. In this work, we identified several PLAs as potent inhibitors of the growth of M. tuberculosis with several of the compounds exhibiting minimum inhibition concentrations equal to or less than 1 μg/mL. Furthermore, two of the compounds were very potent inhibitors of MDR and XDR strains. A high resolution crystal structure of one PLA bound to DHFR from M. tuberculosis reveals the interactions of the ligands with the target enzyme., Competing Interests: The authors have declared that no competing interests exist.

The folate cycle is one of the key metabolic pathways used by bacteria to synthesize vital building blocks required for proliferation. Therapeutic agents targeting enzymes in this cycle, such as trimethoprim and sulfamethoxazole, are among some of the most important and continually used antibacterials to treat both Gram-positive and Gram-negative pathogens. As with all antibacterial agents, the emergence of resistance threatens the continued clinical use of these life-saving drugs. In this article, we describe and analyze resistance mechanisms that have been clinically observed and review newer generations of preclinical compounds designed to overcome the molecular basis of the resistance., (Copyright © 2016 Cold Spring Harbor Laboratory Press; all rights reserved.)

Drug-resistant enzymes must balance catalytic function with inhibitor destabilization to provide a fitness advantage. This sensitive balance, often involving very subtle structural changes, must be achieved through a selection process involving a minimal number of eligible point mutations. As part of a program to design propargyl-linked antifolates (PLAs) against trimethoprim-resistant dihydrofolate reductase (DHFR) from Staphylococcus aureus, we have conducted a thorough study of several clinically observed chromosomal mutations in the enzyme at the cellular, biochemical, and structural levels. Through this work, we have identified a promising lead series that displays significantly greater activity against these mutant enzymes and strains than TMP. The best inhibitors have enzyme inhibition and MIC values near or below that of trimethoprim against wild-type S. aureus. Moreover, these studies employ a series of crystal structures of several mutant enzymes bound to the same inhibitor; analysis of the structures reveals a more detailed molecular understanding of drug resistance in this important enzyme.

The present study examined the efficacy of a short bout of moderately intensive exercise to protect knowledge of a newly acquired motor sequence. Previous work revealed that sleep-dependent offline gains in motor sequence performance are reduced by practicing an alternative motor sequence in close temporal proximity to the original practice with the target motor sequence. In the present work, a brief bout of exercise was inserted at two different temporal locations between practice of a to-be-learned motor sequence and the interfering practice that occurred 2 h later. At issue was whether exposure to exercise could reduce the impact of practice with the interfering task which was expected to be manifest as reemergence of offline gain observed in the case in which the learner is not exposed to the interfering practice. Acute exercise did influence the interfering quality of practice with an alternative motor sequence resulting in the return of broad offline gain. However, this benefit was immediate, emerging on the initial test trial, only when exercise was experienced some time after the original period of motor sequence practice and just prior to practice with the interfering motor sequence. Thus, while exercise can contribute to post-practice consolidation, there appears to be a fragile interplay between spontaneous memory consolidation occurring after task practice and the consolidation processes induced via exercise.

Although classical, negatively charged antifolates such as methotrexate possess high affinity for the dihydrofolate reductase (DHFR) enzyme, they are unable to penetrate the bacterial cell wall, rendering them poor antibacterial agents. Herein, we report a new class of charged propargyl-linked antifolates that capture some of the key contacts common to the classical antifolates while maintaining the ability to passively diffuse across the bacterial cell wall. Eight synthesized compounds exhibit extraordinary potency against Gram-positive S. aureus with limited toxicity against mammalian cells and good metabolic profile. High resolution crystal structures of two of the compounds reveal extensive interactions between the carboxylate and active site residues through a highly organized water network.

Background: Sperm quality may degrade during transit through the male reproductive tract in some individuals. In this setting surgically retrieved testicular sperm may outperform ejaculated samples for use with in vitro fertilization (IVF) and intracytoplasmic sperm injection (IVF-ICSI). We sought to describe one center's experience with the use of fresh testicular sperm after prior failed IVF-ICSI with ejaculated samples., Results: A retrospective review was conducted evaluating IVF-ICSI cycles performed at a tertiary IVF unit between 2009 and 2014. Couples who were managed with percutaneous testis biopsy to obtain sperm, despite availability of ejaculated sperm, were included. Four couples who underwent a total of 6 percutaneous testis biopsy/IVF-ICSI cycles were identified. Collectively, the couples had undergone 9 prior IVF-ICSI cycles using fresh ejaculated sperm without successful pregnancy. From the six cycles that used fresh testicular sperm four live births resulted (1 twin gestation, 3 singletons). Only 1 of the 4 couples remained childless., Conclusions: For patients who have had prior failed IVF-ICSI attempts, this small case series demonstrates a possible therapeutic benefit when freshly procured testicular sperm are used in lieu of ejaculated samples.

Objective: In an effort to better incorporate precision medicine into clinical practice, we initiated a pilot project to screen, discuss, and genetically characterize patients with metastatic or recurrent gynecologic malignancies for whom no curative standard of care exists., Methods: In 7/2014, we initiated a multi-disciplinary Precision Medicine Board (PMB) whose purpose was to apply molecular profiling to select and prioritize early phase clinical trial enrollment for high-risk gynecologic malignancies. Additional objectives were to record outcomes and enable scientific discussions of mutations which may foster local translational research. FoundationOne was the preferred genomic platform; results were reviewed by a team comprised of disease site specialists, phase I trialists, and basic and translational scientists affiliated with the Gynecologic Cancer Program. A detailed database for each patient was created and is followed prospectively for treatment use and resultant outcomes., Results: To date, we have presented 62 cases with interpretable FoundationOne testing on 60 tumor samples (31 ovarian, 18 uterine, 9 cervical, and 4 other female genital tract). Significant genomic alterations were commonly found in all tumor types (median: 3); TP53 (45%) and PIK3CA (27%) were the most frequently noted mutations; however, molecular profiling resulted in identification of few actionable mutations (6%). To date, we have matched 4 patients on therapies based on actionable mutations., Conclusions: The predominant function of our PMB is establishment of a forum to enhance research while providing clinical care for refractory malignancies. We have matched patients with specific mutations to ongoing trials and are developing investigator-initiated studies based on trends within genomic profiling results. Longer-term follow up will be required to determine the success of this strategy., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Study Question: Is dairy food consumption associated with live birth among women undergoing infertility treatment?, Summary Answer: There was a positive association between total dairy food consumption and live birth among women ≥35 years of age., What Is Known Already: Dairy food intake has been previously related to infertility risk and measures of fertility potential but its relation to infertility treatment outcomes are unknown., Study Design, Size, Duration: Our study population comprised a total of 232 women undergoing 353 in vitro fertilization (IVF) treatment cycles between February 2007 and May 2013, from the Environment and Reproductive Health study, an ongoing prospective cohort., Participants/materials, Setting, Methods: Diet was assessed before assisted reproductive technology (ART) treatment using a validated food frequency questionnaire. Study outcomes included ovarian stimulation outcomes (endometrial thickness, estradiol levels and oocyte yield), fertilization rates, embryo quality measures and clinical outcomes (implantation, clinical pregnancy and live birth rates). We used generalized linear mixed models with random intercepts to account for multiple ART cycles per woman while simultaneously adjusting for age, caloric intake, BMI, race, smoking status, infertility diagnosis, protocol type, alcohol intake and dietary patterns., Main Results and the Role of Chance: The age- and calorie-adjusted difference in live birth between women in the highest (>3.0 servings/day) and lowest (<1.34 servings/day) quartile of dairy intake was 21% (P = 0.02). However, after adjusting for additional covariates, this association was observed only among women ≥35 years (P, interaction = 0.04). The multivariable-adjusted live birth (95% CI) in increasing quartiles of total dairy intake was 23% (11, 42%), 39% (24, 56%), 29% (17, 47%) and 55% (39, 69%) (P, trend = 0.02) among women ≥35 years old, and ranged from 46 to 54% among women <35 years old (P, trend = 0.69). There was no association between dairy intake and any of the intermediate outcomes., Limitations, Reasons for Caution: The lack of a known biological mechanism linking dairy intake to infertility treatment outcomes calls for caution when interpreting these results and for additional work to corroborate or refute them., Wider Implications of the Findings: Dairy intake does not appear to harm IVF outcomes and, if anything, is associated with higher chances of live birth., Study Funding/competing Interests: This work was supported by NIH grants R01-ES009718 and R01ES000002 from NIEHS, P30 DK046200 from NIDDK and T32HD060454 from NICHD. M.C.A. was supported by a Ruth L. Kirschstein National Research Service Award T32 DK 007703-16 from NIDDK. She is currently employed at the Nestlé Research Center, Switzerland and completed this work while at the Harvard School of Public Health. The other authors declare no conflicts of interest., (© The Author 2016. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Multidrug-resistant Enterobacteriaceae, notably Escherichia coli and Klebsiella pneumoniae, have become major health concerns worldwide. Resistance to effective therapeutics is often carried by class I and II integrons that can confer insensitivity to carbapenems, extended spectrum β-lactamases, the antifolate trimethoprim, fluoroquinolones, and aminoglycosides. Specifically of interest to the study here, a prevalent gene (dfrA1) coding for an insensitive dihydrofolate reductase (DHFR) confers 190- or 1000-fold resistance to trimethoprim for K. pneumoniae and E. coli, respectively. Attaining inhibition of both the wild-type and resistant forms of the enzyme is critical for new antifolates. For several years, we have been developing the propargyl-linked antifolates (PLAs) as effective inhibitors against trimethoprim-resistant DHFR enzymes. Here, we show that the PLAs are active against both the wild-type and DfrA1 DHFR proteins. We report two high-resolution crystal structures of DfrA1 bound to potent PLAs. The structure-activity relationships and crystal structures will be critical in driving the design of broadly active inhibitors against wild-type and resistant DHFR.

In order to determine how participants represent practiced, discrete keying sequences in the discrete sequence production task, we had 24 participants practice two six-key sequences on the basis of two pre-learned six-digit numbers. These sequences were carried out by fingers of the left (L) and right (R) hand with between-hand transitions always occurring between the second and third, and the fifth and sixth responses. This yielded the so-called LLRRRL and RRLLLR sequences. Early and late in practice, the keypad used for the right hand was briefly relocated from the front of the participants to 90° at their right side. The results indicate that after 600 practice trials, executing a keying sequence relies heavily on a spatial cross-hand representation in a trunk- or head-based reference frame that after about only 15 trials is fully adjusted to the changed hand location. The hand location effect was not found with the last sequence element. This is attributed to the application of explicit knowledge. The between-hand transitions appeared to induce initial segmentation in some of the participants, but this did not consolidate into a concatenation point of successive motor chunks.

Robust offline gains occur during explicit motor sequence learning. However practice of a motor sequence, other than the target sequence, within 4-6 hr after initial practice interferes with these gains. This work assessed if experiencing supplemental practice of spatially or motorically similar sequences influenced the extent of offline gain. A contemporary model of sequence learning assumes that exposure to a spatially but not motor-related sequence would not compromise stabilization of the memory for the target sequence, thus, would have minimal impact on the resultant offline gain and possibly amplify overnight gains. As anticipated, a reliable offline improvement was observed for the target motor sequence in the absence of practice with an alternative motor sequence. This gain was significantly reduced when the learner experienced additional practice with either a novel or motorically similar sequence. There was no evidence of heightened overnight gain for the target sequence from intervening practice with a spatially similar sequence. Thus, the expression of offline improvement is not necessarily eliminated if practice of an alternative motor sequence is encountered shortly after a target sequence. However, the structure of the motor sequence faced during intervening practice can impact the resultant postpractice consolidation processes reflected in the eventual expression of offline facilitation.

Two experiments were conducted to examine the efficacy of random (RP) and blocked practice (BP) for enhancing later motor learning. Each experiment involved practicing three unique seven key serial reaction time (SRT) tasks in either a blocked or random format followed by practice of a novel SRT task either 2-min (Experiment 1) or 24-h (Experiment 2) later. While the expected benefit of RP for retention was present in both experiments, in Experiment 1 there was no advantage from prior RP for new learning. Experiment 2 explored the possibility that increasing the interval, from 2-min to 24-h, between BP or RP and practice of the novel motor task might allow consolidation of sequence knowledge acquired during BP or RP which in turn might facilitate new learning. As a result of the additional time between training bouts RP facilitated the rate at which the novel motor task was acquired. Interestingly, when this additional time was provided, both BP and RP supported (a) a performance saving for the first trial with the novel task, and (b) an offline improvement in performance across a 24-h interval not present when only the novel motor task was practiced. The latter benefits for new learning may have resulted from exposure to prior physical practice per se. or practice variability. These data are discussed with respect to (a) future learning benefits from prior experience training with greater CI, and (b) the importance of memory consolidation for motor learning., (Published by Elsevier B.V.)

We performed two experiments to determine whether amplified motor output variability and compromised processing of visual information in older adults impair short-term adaptations when learning novel fine motor tasks. In Experiment 1, 12 young and 12 older adults underwent training to learn how to accurately trace a sinusoidal position target with abduction-adduction of their index finger. They performed 48 trials, which included 8 blocks of 6 trials (the last trial of each block was performed without visual feedback). Afterward, subjects received an interference task (watched a movie) for 60 min. We tested retention by asking subjects to perform the sinusoidal task (5 trials) with and without visual feedback. In Experiment 2, 12 young and 10 older adults traced the same sinusoidal position target with their index finger and ankle at three distinct visual angles (0.25°, 1° and 5.4°). In Experiment 1, the movement error and variability were greater for older adults during the visual feedback trials when compared with young adults. In contrast, during the no-vision trials, age-associated differences in movement error and variability were ameliorated. Short-term adaptations in learning the sinusoidal task were similar for young and older adults. In Experiment 2, lower amount of visual feedback minimized the age-associated differences in movement variability for both the index finger and ankle movements. We demonstrate that although short-term adaptations are similar for young and older adults, older adults do not process visual information as well as young adults and that compromises their ability to control novel fine motor tasks during acquisition, which could influence long-term retention and transfer.

In 1999, crop consultants scouting for stink bugs (Hemiptera spp.) in South Carolina discovered a formerly unobserved seed rot of cotton that caused yield losses ranging from 10 to 15% in certain fields. The disease has subsequently been reported in fields throughout the southeastern Cotton Belt. Externally, diseased bolls appeared undamaged; internally, green fruit contain pink to dark brown, damp, deformed lint, and necrotic seeds. In greenhouse experiments, we demonstrated transmission of the opportunistic bacterium Pantoea agglomerans by the southern green stink bug, Nezara viridula (L.). Here, green bolls were sampled from stink bug management plots (insecticide protected or nontreated) from four South Atlantic coast states (North Carolina, South Carolina, Georgia, and Florida) to determine disease incidence in the field and its association with piercing-sucking insects feeding. A logistic regression analysis of the boll damage data revealed that disease was 24 times more likely to occur (P = 0.004) in bolls collected from plots in Florida, where evidence of pest pressure was highest, than in bolls harvested in NC with the lowest detected insect pressure. Fruit from plots treated with insecticide, a treatment which reduced transmission agent numbers, were 4 times less likely to be diseased than bolls from unprotected sites (P = 0.002). Overall, punctured bolls were 125 times more likely to also have disease symptoms than nonpunctured bolls, irrespective of whether or not plots were protected with insecticides (P = 0.0001). Much of the damage to cotton bolls that is commonly attributed to stink bug feeding is likely the resulting effect of vectored pathogens., (Published by Oxford University Press on behalf of Entomological Society of America 2015. This work is written by US Government employees and is in the public domain in the US.)

While antifolates such as Bactrim (trimethoprim-sulfamethoxazole; TMP-SMX) continue to play an important role in treating community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA), resistance-conferring mutations, specifically F98Y of dihydrofolate reductase (DHFR), have arisen and compromise continued use. In an attempt to extend the lifetime of this important class, we have developed a class of propargyl-linked antifolates (PLAs) that exhibit potent inhibition of the enzyme and bacterial strains. Probing the role of the configuration at the single propargylic stereocenter in these inhibitors required us to develop a new approach to nonracemic 3-aryl-1-butyne building blocks by the pairwise use of asymmetric conjugate addition and aldehyde dehydration protocols. Using this new route, a series of nonracemic PLA inhibitors was prepared and shown to possess potent enzyme inhibition (IC50 values <50 nM), antibacterial effects (several with MIC values <1 μg/mL) and to form stable ternary complexes with both wild-type and resistant mutants. Unexpectedly, crystal structures of a pair of individual enantiomers in the wild-type DHFR revealed that the single change in configuration of the stereocenter drove the selection of an alternative NADPH cofactor, with the minor α-anomer appearing with R-27. Remarkably, this cofactor switching becomes much more prevalent when the F98Y mutation is present. The observation of cofactor site plasticity leads to a postulate for the structural basis of TMP resistance in DHFR and also suggests design strategies that can be used to target these resistant enzymes.

The pathogen causing soybean rust, Phakopsora pachyrhizi, was first described in Japan in 1902. The disease was important in the Eastern Hemisphere for many decades before the fungus was reported in Hawaii in 1994, which was followed by reports from countries in Africa and South America. In 2004, P. pachyrhizi was confirmed in Louisiana, making it the first report in the continental United States. Based on yield losses from countries in Asia, Africa, and South America, it was clear that this pathogen could have a major economic impact on the yield of 30 million ha of soybean in the United States. The response by agencies within the United States Department of Agriculture, industry, soybean check-off boards, and universities was immediate and complex. The impacts of some of these activities are detailed in this review. The net result has been that the once dreaded disease, which caused substantial losses in other parts of the world, is now better understood and effectively managed in the United States. The disease continues to be monitored yearly for changes in spatial and temporal distribution so that soybean growers can continue to benefit by knowing where soybean rust is occurring during the growing season.

We report the first Raman spectroscopic study of propargyl-linked dihydrofolate reductase (DHFR) inhibitors being taken up by wild type Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus cells. A novel protocol is developed where cells are exposed to the fermentation medium containing a known amount of an inhibitor. At a chosen time point, the cells are centrifuged and washed to remove the extracellular compound, then frozen and freeze-dried. Raman difference spectra of the freeze-dried cells (cells exposed to the drug minus cells alone) provide spectra of the compounds inside the cells, where peak intensities allow us to quantify the number of inhibitors within each cell. A time course for the propargyl-linked DHFR inhibitor UCP 1038 soaking into E. coli cells showed that penetration occurs very quickly and reaches a plateau after 10 min exposure to the inhibitor. After 10 min drug exposure, the populations of two inhibitors, UCP 1038 and UCP 1089, were ~1.5 × 10(6) molecules in each E. coli cell, ~4.7 × 10(5) molecules in each K. pneumonia cell, and ~2.7 × 10(6) in each S. aureus cell. This is the first in situ comparison of inhibitor population in Gram-negative and Gram-positive bacterial cells. The positions of the Raman peaks also reveal the protonation of diaminopyrimidine ring upon binding to DHFR inside cells. The spectroscopic signature of protonation was characterized by binding an inhibitor to a single crystal of DHFR.

Purpose: To compare the in-vitro fertilization (IVF) outcomes of cancer patients who underwent oocyte retrieval and embryo/oocyte cryopreservation prior to gonadotoxic therapy to those of age and time-matched controls with tubal factor infertility., Methods: All cancer patients who underwent embryo/oocyte cryopreservation at our institution from 1997 to 2014 were reviewed. Primary outcomes were total dose of gonadotropins used, number of oocytes retrieved, and number of 2pn embryos obtained. Outcomes were compared to age-matched controls with tubal-factor infertility who underwent a fresh embryo transfer within the same relative time period as the IVF cycle of the cancer patient., Results: Sixty-three cancer patients underwent 65 IVF cycles, and 21 returned for frozen embryo transfer. One hundred twenty-two age-matched controls underwent IVF cycles with fresh transfer, and 23 returned for frozen embryo transfer. No difference was seen between cancer patients and controls with respect to total ampules of gonadotropin used (38.0 vs. 35.6 respectively; p = 0.28), number of oocytes retrieved (12.4 vs. 10.9 respectively; p = 0.36) and number of 2pn embryos obtained (6.6 vs. 7.1 respectively; p = 0.11). Cumulative pregnancy rate per transfer for cancer patients compared to controls was 37 vs. 43 % respectively (p = 0.49) and cumulative live birth rate per transfer was 30 vs. 32 % respectively (p = 0.85). Cancer patients had a higher likelihood of live birth resulting in twins (44 vs. 14 %; p = 0.035)., Conclusions: Most IVF outcomes appear comparable for cancer patients and age-matched controls. Higher twin pregnancy rates in cancer patients may reflect lack of underlying infertility or need for cancer-specific transfer guidelines.