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The rhizosphere is the region of soil directly influenced by plant roots. The microbial community in the rhizosphere includes fungi, protists, and bacteria, all of which play a significant role in plant health. The beneficial bacteriumSinorhizobium melilotiinfects growing root hairs on nitrogen-starved leguminous plants. Infection leads to the formation of a root nodule, whereS. meliloticonverts atmospheric nitrogen to ammonia, a usable form of nitrogen for plants.S. melilotiis often found in biofilms and travels slowly along the roots, leaving developing root hairs at the growing root tips uninfected. Soil protists are an important component of the rhizosphere system who prey on soil bacteria and have been known to egest undigested phagosomes. We show that the soil protist,Colpoda sp., can transportS. melilotidownMedicago truncatularoots. By using pseudo-3D soil microcosms, we directly observed the presence of fluorescently labelledS. melilotialongM. truncatularoots and tracked the displacement of the fluorescence signal over time. Two weeks after co-inoculation, this signal was detected 52 mm, on average, farther down the roots whenColpoda sp. was also present compared with the experimental treatment that contained bacteria but not protists. Direct counts also showed that protists are required for viable bacteria to reach the deeper sections of root systems in our microcosms. Facilitating bacterial transport may be an important mechanism whereby soil protists promote plant health. As a sustainable agriculture biotechnology, protist-facilitated transport has the potential to boost efficacy of bacterial inoculants, thereby helping growers avoid overuse of nitrogen fertilizers and enhance performance of climate-smart, no-till farming practices.ImportanceSoil protists are an important part of the microbial community in the rhizosphere. Plants grown with protists fare better than plants grown without protists. Mechanisms through which protists support plant health include nutrient cycling, alteration of the bacterial community through selective feeding, and consumption of plant pathogens. Here we provide data in support of an additional mechanism: protists act as transport vehicles for bacteria in soil. We show that protist-facilitated transport can deliver plant-beneficial bacteria to the growing tips of roots that may otherwise be sparsely inhabited with bacteria originating from a seed-associated inoculum. By co-inoculatingMedicago truncatularoots with bothS. meliloti, a nitrogen fixing legume symbiont, andColpoda sp., a ciliated protist, we show substantial and statistically significant transport with depth and breadth of bacteria-associated fluorescence as well as transport of viable bacteria. Co-inoculation with shelf-stable encysted soil protists may be employed as a sustainable agriculture biotechnology to better distribute beneficial bacteria and enhance the performance of inoculants.

[Image: see text] Allosteric modulation of GPCRs represents an increasingly explored approach in drug development. Due to complex pharmacology, however, the relationship(s) between modulator properties determined in vitro with in vivo concentration-effect phenomena is frequently unclear. We investigated key pharmacological properties of a set of metabotropic glutamate receptor 5 (mGlu(5)) positive allosteric modulators (PAMs) and their relevance to in vivo concentration–response relationships. These studies identified a significant relationship between in vitro PAM cooperativity (αβ), as well as the maximal response obtained from a simple in vitro PAM concentration–response experiment, with in vivo efficacy for reversal of amphetamine-induced hyperlocomotion. This correlation did not exist with PAM potency or affinity. Data across PAMs were then converged to calculate an in vivo concentration of glutamate putatively relevant to the mGlu(5) PAM mechanism of action. This work demonstrates the ability to merge in vitro pharmacology profiles with relevant behavioral outcomes and also provides a novel method to estimate neurotransmitter concentrations in vivo.

Both quantitative electroencephalography (qEEG) and cognitive measures have been used to understand the underlying brain changes that occur in individuals after experiencing a traumatic brain injury, however, research exploring the relationship between qEEG patterns and cognitive test performance is scarcely studied in school-aged populations. The purpose of the present study was to explore first, the neuropsychological and academic deficits in young individuals with TBI; and second, the underlying relationship between qEEG patterns and cognitive test performance. Analyses included 21 school-aged participants whom have experienced a recent TBI and 15 school-aged participants whom have never experienced a TBI. Mean subtest and composite scores were compared and regression analyses were used to determine whether alpha band and beta band qEEG coherence values predicted processing speed measures. Results suggest that young individuals who experienced a recent TBI exhibit general deficits in cognition and academic skills beyond what would be expected in the general population. Further, beta band coherence with the frontal brain regions significantly predicted processing speed scores, providing evidence of a relationship between qEEG patterns and processing speed. This outlines a relatively inexpensive method for utilizing neural connectivity to verify cognitive deficits for school-aged individuals with a recent TBI.

Over-seeding cool-season annual forages in pastures in the southeastern US provides increased grazing opportunities for livestock and reduces the amount of hay needed during winter. The constant presence of vegetation can benefit soil health, the biological aspect of which has only received cursory investigation to date. Since 2009, a commercial cattle grazing operation located in south Mississippi has been over-seeding a diverse mixture of oats (Avena sativa L.), triticale (X Triticosecale Wittmack), annual ryegrass (Lolium multiflorum Lam.), hairy vetch (Vicia villosa Roth), radish (Raphanus sativus L.), turnips (Brassica rapa L. subsp. rapa), red clover (Trifolium pratense L.), ball clover (Trifolium nigrescens Viv.), and crimson clover (Trifolium incarnatum L.) into pastures dominated by bahiagrass (Paspalum notatum Flugge). Soil health properties were assessed across a heterogeneous landscape from soils collected immediately prior to seeding and following increased spring productivity of cool-season annuals from 2015 to 2017. Samples were collected across a topographic sequence to a depth of 15 and 30 cm. Three samples per site were collected from summit, backslope, and footslope locations. Soil chemical assessments included macronutrients, pH, soil organic matter (SOM), total carbon (TC), and total nitrogen (TN). Fatty acid methyl ester analysis and enzyme assays were used to determine soil microbial community structure and activity. While TC and TN did not change with time (20.5 and 2.4 g kg−1, respectively), SOM increased by 6% from 2015 to 2017. Nitrate-N decreased by 82% from 2015 to 2017. N-acetyl-β-glucosaminidase activity increased by 32%, while β-glucosidase remained constant. Total microbial abundance did not change with time; however, Gram positive bacteria and actinomycetes decreased by 51% and 43%, respectively, from fall to spring. Arbuscular mycorrhizal fungi increased from fall to spring by an average of 45%. This pasture management technique of over-seeding a diverse mix of winter annuals may have resulted in a steady state of TC and TN, while SOM increased and inorganic nitrogen decreased with time, and enhanced the total soil microbial community particularly fungi and their potential enzyme activity. Despite some biological indications of system stability, soil properties potentially limiting to plant growth were identified for some sites, but these limitations were not closely related to topographic position as expected.

This letter describes progress towards an M4 PAM preclinical candidate inspired by an unexpected aldehyde oxidase (AO) metabolite of a novel, CNS penetrant thieno[2,3-c]pyridine core to an equipotent, non-CNS penetrant thieno[2,3-c]pyrdin-7(6H)-one core. Medicinal chemistry design efforts yielded two novel tricyclic cores that enhanced M4 PAM potency, regained CNS penetration, displayed favorable DMPK properties and afforded robust in vivo efficacy in reversing amphetamine-induced hyperlocomotion in rats.

Opioid use disorder (OUD) is a debilitating neuropsychiatric condition characterized by compulsive opioid use, dependence, and repeated relapse after periods of abstinence. Given the high risk of d...

This letter describes progress towards an M4 PAM preclinical candidate that resulted in the discovery of VU6005806/AZN-00016130. While the thieno[2,3-c]pyridazine core has been a consistent feature of key M4 PAMs, no work had previously been reported with respect to alternate functionality at the C3 position of the pyridazine ring. Here, we detail new chemistry and analogs that explored this region, and quickly led to VU6005806/AZN-00016130, which was profiled as a putative candidate. While, the β-amino carboxamide moiety engendered solubility limited absorption in higher species precluding advancement (or requiring extensive pharmaceutical sciences formulation), VU6005806/AZN-00016130 represents a new, high quality preclinical in vivo probe.

This letter describes a focused, multi-dimensional optimization campaign around BL-1249, a fenamate class non-steroidal anti-inflammatory and a known activator of the K2P potassium channels TREK-1 (K2P2.1) and TREK-2 (K2P10.1). While BL-1249 has been widely profiled in vitro as a dual TREK-1/2 activator, poor physicochemical and DMPK properties have precluded a deeper understanding of the therapeutic potential of these key K2P channels across a broad spectrum of peripheral and central human disease. Here, we report multi-dimensional SAR that led to a novel TREK-1/2 dual activator chemotype, exemplified by ONO-2960632/VU6011992, with improved DMPK properties, representing a new lead for further optimization towards robust in vivo tool compounds.

Objective To describe the known predictors and pathophysiological mechanisms of chronic painful chemotherapy-induced peripheral neuropathy (CIPN) in cancer survivors and the challenges in assessing and managing it. Data Sources PubMed/Medline, CINAHL, Scopus, and PsycINFO. Conclusion The research on chronic painful CIPN is limited. Additional research is needed to identify the predictors and pathophysiological mechanisms of chronic painful CIPN to inform the development of assessment tools and management options for this painful and possibly debilitating condition. Implications for Nursing Practice Recognition of the predictors of chronic painful CIPN and proactive CIPN assessment and palliative management are important steps in reducing its impact on physical function and quality of life.

Objectives Professional athletes undergo annual pre-season laboratory screening, although clinical evidence supporting the practice is limited and no uniform set of guidelines on pre-season laboratory screening exists. The aim of this study was to assess the clinical value of annual pre-season laboratory screening tests for a major professional sports team over multiple years. Design Retrospective chart review. Methods A retrospective analysis was performed of all laboratory results as well as screening ECGs for a single major professional sports team over a 9-year timeframe (2009–2017). Results The data show that 10.01% of initial screening test results were abnormal and 40.32% of abnormal tests resulted in additional testing. Overall, only 0.35% of initial tests resulted in a clinically significant outcome. Non-US born players showed a significantly higher average rate of abnormal tests/year compared to US-born players (p-value 0.006), but there was no difference in clinically significant outcomes. There was no relationship between athlete age and laboratory screening outcomes. Conclusions In our study population, yearly pre-season laboratory screening of professional athletes did not yield substantial clinically significant outcomes and would not be warranted under normal clinical standards. Future best practice guidelines should combine research concerning effects of family medical history, race, gender, country of origin, and type of sport on athlete health when creating recommendations for which pre-season laboratory screenings may be pertinent even with evidence of little utility.

Studies have identified differences between the gut microbiota of colorectal cancer (CRC) patients versus healthy individuals. In this review, we assess the scientific literature to determine if gut microbes should be considered causal, co-varying, or a necessary but not sufficient agent in CRC development. Oral bacteria may influence CRC susceptibility. Colonic biofilms in both sporadic and hereditary CRC suggest these bacteria are present in early neoplasia. Pathogenic drivers and opportunistic passenger bacteria may underlie direct effect of the gut microbiota on carcinogenesis. Members of multiple bacterial taxa have been implicated in CRC tumorigenesis and progression, with distinct mechanisms of action described for each. Individual bacterial organisms found in the colon are likely not enough to explain CRC development and progression. The entire colonic environment, including genetic factors, local tissue inflammatory state as well as dietary components may influence the way epithelial cells respond to the presence of certain bacteria. Longitudinal, human intervention studies are needed to completely clarify complex interactions in the colonic environment and specific causative pathways between the microbiota and CRC.

Individualized feeding care plans and safe handling of milk (human or formula) are critical in promoting growth, immune function, and neurodevelopment in the preterm infant. Feeding errors and disruptions or limitations to feeding processes in the neonatal intensive care unit (NICU) are associated with negative safety events. Feeding errors include contamination of milk and delivery of incorrect or expired milk and may result in adverse gastrointestinal illnesses.The purpose of this review was to evaluate the effect(s) of centralized milk preparation, use of trained technicians, use of bar code-scanning software, and collaboration between registered dietitians and registered nurses on feeding safety in the NICU.A systematic review of the literature was completed, and 12 articles were selected as relevant to search criteria. Study quality was evaluated using the Downs and Black scoring tool.An evaluation of human studies indicated that the use of centralized milk preparation, trained technicians, bar code-scanning software, and possible registered dietitian involvement decreased feeding-associated error in the NICU.A state-of-the-art NICU includes a centralized milk preparation area staffed by trained technicians, care supported by bar code-scanning software, and utilization of a registered dietitian to improve patient safety. These resources will provide nurses more time to focus on nursing-specific neonatal care.Further research is needed to evaluate the impact of factors related to feeding safety in the NICU as well as potential financial benefits of these quality improvement opportunities.

This letter describes a focused exercise to explore the role of the β-amino carboxamide moiety found in all of the first generation M4 PAMs and question if the NH2 group served solely to stabilize an intramolecular hydrogen bond (IMHB) and enforce planarity. To address this issue (and to potentially find a substitute for the β-amino carboxamide that engendered P-gp and contributed to solubility liabilities), we removed the NH2, generating des-amino congeners and surveyed other functional groups in the β-position. These modifications led to weak M4 PAMs with poor DMPK properties. Cyclization of the β-amino carboxamide moiety by virtue of a pyrazole ring re-enforced the IMHB, led to potent (and patented) M4 PAMs, many as potent as the classical bicyclic β-amino carboxamide analogs, but with significant CYP1A2 inhibition. Overall, this exercise indicated that the β-amino carboxamide moiety most likely facilitates an IMHB, and is essential for M4 PAM activity within classical bicyclic M4 PAM scaffolds.

Fast food consumption is linked to poor health, yet many older adults regularly consume fast food. Understanding factors contributing to fast food consumption is useful in the development of targeted interventions. The aim of this study was to characterize how fast food consumption relates to socio-demographic characteristics in a low-income sample of older adults.This study used cross-sectional survey data of 50 to79-year-olds (N-236) in urban safety-net clinics in 2010 in Kansas City, KS. Self-reported frequency of fast food consumption was modeled using ordinal logistic regression with socio-demographics as predictor variables. Participants were 56.8 ± 6.0 (mean ± SD) years old, 64% female, 45% non-Hispanic African American, and 26% Hispanic. Thirty-nine percent denied eating fast food in the past week, 36% ate once, and 25% ate fast food at least twice. Age was negatively correlated with fast food intake (r = −0.20, P = 0.003). After adjusting for age, race-ethnicity, employment, and marital status, the association between education and fast food consumption differed by sex (Pinteraction = 0.017). Among women, higher education was associated with greater fast food intake (Spearman's correlation; r = 0.28, P = 0.0005); the association was not significant in men (r = −0.14, P = 0.21). In this diverse, low-income population, high educational attainment (college graduate or higher) related to greater fast food intake among women but not men. Exploration of the factors contributing to this difference could inform interventions to curb fast food consumption or encourage healthy fast food choices among low-income, older adults. Keywords: Fast food, Older adults, Diet quality, Education, Socio-demographic characteristics

The nonstructural protein 1 (nsp1) of severe acute respiratory syndrome coronavirus and severe acute respiratory syndrome coronavirus 2 is a critical viral protein that suppresses host gene expression by blocking the assembly of the ribosome on host mRNAs. To understand the mechanism of inhibition of host gene expression, we sought to identify cellular proteins that interact with nsp1. Using proximity-dependent biotinylation followed by proteomic analyses of biotinylated proteins, here we captured multiple dynamic interactions of nsp1 with host cell proteins. In addition to ribosomal proteins, we identified several pre-mRNA processing proteins that interact with nsp1, including splicing factors and transcription termination proteins, as well as exosome, and stress granule (SG)-associated proteins. We found that the interactions with transcription termination factors are primarily governed by the C-terminal region of nsp1 and are disrupted by the mutation of amino acids K164 and H165 that are essential for its host shutoff function. We further show that nsp1 interacts with Ras GTPase-activating protein SH3 domain-binding protein 1 (G3BP1) and colocalizes with G3BP1 in SGs under sodium arsenite-induced stress. Finally, we observe that the presence of nsp1 disrupts the maturation of SGs over a long period. Isolation of SG core at different times shows a gradual loss of G3BP1 in the presence of nsp1.

Juvenile idiopathic arthritis (JIA) is a chronic inflammatory disease affecting the joints and other organs that occurs in 1 in 1,000 children in the United States. Given the various categories of JIA, interpretation of the literature can be difficult. In this review, new developments in understanding non-systemic JIA and its treatment will be covered. Recent advances in the journey toward personalized treatment in JIA will be highlighted, including a review of currently available biologic modifiers. Uveitis and the temporomandibular joint will be discussed as particularly challenging treatment issues. Recent guideline updates and literature-guided treatment decisions will be reviewed.

Dysgonomonas species are facultative heterotrophs capable of growth on lignocellulose-derived polysaccharides. Dysgonomonas species harbor myriad genes involved in glycan modification and are well suited to the lignocellulose-rich conditions within the termite hindgut. Here, we report draft genome sequences for Dysgonomonas sp. strains GY75 and GY617, isolated from the hindgut of Reticulitermes flavipes.

Dysgonomonas spp. are facultative heterotrophs which colonize diverse environments, including the hindgut of the lower termite Reticulitermes flavipes . Dysgonomonas genomes are enriched for genes involving oligo- and polysaccharide utilization, enabling modification of a wide array of complex glycans. Here, we report draft genome sequences for Dysgonomonas sp. strains BGC7 and HGC4.

Microorganisms play a vital role in shaping the soil environment and enhancing plant growth by interacting with plant root systems. Because of the vast diversity of cell types involved, combined with dynamic and spatial heterogeneity, identifying the causal contribution of a defined factor, such as a microbial exopolysaccharide (EPS), remains elusive. Synthetic approaches that enable orthogonal control of microbial pathways are a promising means to dissect such complexity. Here we report the implementation of a synthetic, light-activated, transcriptional control platform using the blue-light responsive DNA binding protein EL222 in the nitrogen fixing soil bacterium

A neuropsychological interpretive schema is advantageous in its ability to provide a framework for understanding the potential role of internal information processing capacities in causing learning problems. Neuropsychology provides a roadmap for understanding potential cognitive causes of behavior and is an imperative step for understanding learning problems in children. The current chapter explores how neuropsychology can be applied in education settings. Neuropsychology has had a significant influence on disability evaluations of children in special education. Despite its direct relevance for understanding children with neurodevelopmental disabilities, neuropsychological research and training have been highly under-represented in the field of school psychology. There are several barriers to its inclusion in the field including a lack of training, personnel, and theoretical biases. Furthermore, a lack of simplified models of brain functioning that provide clear guidance for evaluations, specifically intended for educational purposes, is a major obstacle for the growth of neuropsychology in the school setting. Additionally, school psychologists frequently criticize the interpretive component of neuropsychology since it appears to involve a high degree of inference on the part of the assessor in order to provide a conclusive diagnosis. The current chapter addresses these issues by (1) providing outlines and examples for those wishing to implement neuropsychology in educational assessments, (2) introducing a new method for interpreting psychoeducational assessments that is more explicit and objective than clinical judgment in isolation, and (3) providing direct guidance for possible intervention approaches.

Background: Chemotherapy-induced peripheral neuropathy (CIPN) is commonly experienced by children receiving neurotoxic chemotherapy. No validated pediatric CIPN patient-reported outcome (PRO) measures exist. Purpose: To test sensitivity, internal consistency reliability, content and convergent validity, and feasibility of the Pediatric Chemotherapy-Induced Neuropathy (P-CIN), an electronic PRO measure for assessing CIPN in children who received neurotoxic chemotherapy. Method: Five experts evaluated content validity of the 14-item P-CIN. Children 5 to 17 years old with CIPN ( N = 79) completed the P-CIN via tablet computer; a subset ( n = 26) also underwent neurological examinations using the Pediatric–Modified Total Neuropathy Score. Following preliminary analyses, one item was deleted and three others modified. The revised P-CIN was retested with patients ( n = 6) who also completed the Bruininks–Oseretsky Test of Motor Proficiency motor function assessment. Means, item response ranges, standard deviations, content validity indexes, Cronbach’s alphas, and correlation coefficients were calculated. Results: Mean participant age was 11.25 ( SD = 4.0) years. Most had acute leukemia (62.5%) and received vincristine (98.7%). Content validity index coefficients ranged from .80 to 1.0 ( p = .05). For 9 of 14 items, responses ranged from 0 to 4 or 5; response ranges for toe numbness, pick up a coin, and three of four pain items were 0 to 3. After deleting one item, Cronbach’s alpha coefficient was .83. P-CIN scores were strongly associated with Pediatric-Modified Total Neuropathy Score ( r = .52, p < .01) and Bruininks–Oseretsky Test of Motor Proficiency ( r = −.83, p = .04) scores. Sixty-eight percent of children 6 to 17 years old completed P-CIN independently. Discussion: Preliminary evidence suggests that the 13-item P-CIN is internally consistent, is valid, and can be completed independently by children ≥ 6 years. However, we recommend additional testing.

Objective In the U.S., youths account for most annual mild traumatic brain injury (mTBI) cases. Often mTBI patients experience impairing symptoms that affect several domains of functioning, typically these symptoms remit within a month though many patients will experience impairing symptoms that persist for greater than three months. Due to the current difficulties in diagnosing this condition in a standardized way, the current study aims to clarify the cognitive profiles of persistent post-concussion syndrome (PPCS) in youth patients. Method 30 youths between the ages of 8 and 18 years participated. 15 youths with mTBI symptoms for greater than three months were recruited from a mTBI treatment clinic. An additional 15 youths without a mTBI history were recruited as a control group. All participants underwent cognitive testing using the standard Woodcock-Johnson Tests of Cognitive Abilities battery. Logistic regression analyses explored the importance of cognitive processing speed and working memory for identifying youth patients with PPCS. Further exploratory analyses were conducted to clarify additional cognitive domains that may be impacted by PPCS. Results Findings suggest processing speed and working memory abilities alone are not adequate for identifying patients with PPCS and measures of processing speed, fluid reasoning, working memory, and long-term retrieval may be important to consider when differentially diagnosing PPCS. Conclusions More research is needed to appropriately identify and validate diagnostic practices for PPCS in youth. Our findings provide a basis for future work and suggests neuropsychological batteries that include measures for these four cognitive domains may be valuable for identifying PPCS in youth patients.

Non-selective antagonists of muscarinic acetylcholine receptors (mAChRs) that broadly inhibit all five mAChR subtypes provide an efficacious treatment for some movement disorders, including Parkinson disease and dystonia. Despite their efficacy in these and other central nervous system disorders, anti-muscarinic therapy has limited utility due to severe adverse effects that often limit their tolerability by patients. Recent advances in understanding the roles that each mAChR subtype plays in disease pathology suggest that highly selective ligands for individual subtypes may underlie the anti-parkinsonian and anti-dystonic efficacy observed with the use of non-selective anti-muscarinic therapeutics. Our recent work has indicated that the M4muscarinic acetylcholine receptor has several important roles in opposing aberrant neurotransmitter release, intracellular signaling pathways, and brain circuits associated with movement disorders. This raises the possibility that selective antagonists of M4may recapitulate the efficacy of non-selective anti-muscarinic therapeutics and may decrease or eliminate the adverse effects associated with these drugs. However, this has not been directly tested due to lack of selective antagonists of M4. Here we utilize genetic mAChR knockout animals in combination with non-selective mAChR antagonists to confirm that the M4receptor underlies the locomotor-stimulating and anti-parkinsonian efficacy in rodent models. We also report the synthesis, discovery, and characterization of the first-in-class selective M4antagonists VU6013720, VU6021302, and VU6021625 and confirm that these optimized compounds have anti-parkinsonian and anti-dystonic efficacy in pharmacological and genetic models of movement disorders.

[Image: see text] Nonselective antagonists of muscarinic acetylcholine receptors (mAChRs) that broadly inhibit all five mAChR subtypes provide an efficacious treatment for some movement disorders, including Parkinson’s disease and dystonia. Despite their efficacy in these and other central nervous system disorders, antimuscarinic therapy has limited utility due to severe adverse effects that often limit their tolerability by patients. Recent advances in understanding the roles that each mAChR subtype plays in disease pathology suggest that highly selective ligands for individual subtypes may underlie the antiparkinsonian and antidystonic efficacy observed with the use of nonselective antimuscarinic therapeutics. Our recent work has indicated that the M(4) muscarinic acetylcholine receptor has several important roles in opposing aberrant neurotransmitter release, intracellular signaling pathways, and brain circuits associated with movement disorders. This raises the possibility that selective antagonists of M(4) may recapitulate the efficacy of nonselective antimuscarinic therapeutics and may decrease or eliminate the adverse effects associated with these drugs. However, this has not been directly tested due to lack of selective antagonists of M(4). Here, we utilize genetic mAChR knockout animals in combination with nonselective mAChR antagonists to confirm that the M(4) receptor activation is required for the locomotor-stimulating and antiparkinsonian efficacy in rodent models. We also report the synthesis, discovery, and characterization of the first-in-class selective M(4) antagonists VU6013720, VU6021302, and VU6021625 and confirm that these optimized compounds have antiparkinsonian and antidystonic efficacy in pharmacological and genetic models of movement disorders.

Members of Dysgonomonas are Gram-stain-negative, non-motile, facultatively anaerobic coccobacilli originally described in relation to their isolation from stool and wounds of human patients (CDC group DF-3). More recently, Dysgonomonas have been found to be widely distributed in terrestrial environments and are particularly enriched in insect systems. Their prevalence in xylophagous insects such as termites and wood-feeding cockroaches, as well as in soil-fed microbial fuel cells, elicit interest in lignocellulose degradation and biofuel production, respectively. Their occurrence in mosquito and fruit fly have implications relating to symbiosis, host immunology and developmental biology. Additionally, their presence in termite, mosquito and nematode present novel opportunities for pest and vector control. Currently, the absolute growth requirements of Dysgonomonas are unknown, and they are commonly cultured under anaerobic conditions on complex media containing blood, peptones, tryptones, and yeast, plant or meat extracts. Restrictive and undefined culturing conditions preclude physiological and genetic studies, and thus further understanding of their metabolic potential. Here we describe the requirements for growth of termite-derived Dysgonomonas isolates and create parallel complex, defined and minimal media that permit vigorous and reliable aerobic growth. Furthermore, we show that these media can be used to easily enrich for Dysgonomonas isolates from densely-colonized and microbially-diverse environmental samples.

Objective To determine the effectiveness of liquid human milk fortifiers (HMFs) derived from exclusive HM or hydrolyzed protein on growth, necrotizing enterocolitis (NEC), or late-onset sepsis in North American very low-birth-weight (VLBW) infants compared with powder HMFs (control). Methods Prospective trials published between 2009 and 2020 were systematically reviewed, and meta-analysis was conducted by using a random-effects model. Results Five studies were identified for up to 591 participants across 39 centers. Study treatments included whey or casein hydrolysate HMF and exclusive HM HMF. Infants fed whey or casein hydrolysate HMF had growth differences compared with the control. No differences were found across treatments in regard to NEC or sepsis. Conclusion Very low-quality evidence suggests greater linear growth in VLBW infants fed whey hydrolysate liquid HMF, as well as greater weight gain in those fed casein hydrolysate HMF, compared with the control. Additional prospective, multicenter randomized controlled trials are needed to confirm these estimates because of sparsity of evidence. There is insufficient evidence to support HMF decisions regarding NEC or late-onset sepsis prophylaxis.

Degeneration of basal forebrain cholinergic circuitry represents an early event in the development of Alzheimer’s disease (AD). These alterations in central cholinergic function are associated with disruptions in arousal, sleep/wake architecture, and cognition. Changes in sleep/wake architecture are also present in normal aging and may represent a significant risk factor for AD. M(1) muscarinic acetylcholine receptor (mAChR) positive allosteric modulators (PAMs) have been reported to enhance cognition across preclinical species and may also provide beneficial effects for age- and/or neurodegenerative disease-related changes in arousal and sleep. In the present study, electroencephalography was conducted in young animals (mice, rats and nonhuman primates [NHPs]) and in aged mice to examine the effects of the selective M(1) PAM VU0453595 in comparison with the acetylcholinesterase inhibitor donepezil, M(1)/M(4) agonist xanomeline (in NHPs), and M(1) PAM BQCA (in rats) on sleep/wake architecture and arousal. In young wildtype mice, rats, and NHPs, but not in M(1) mAChR KO mice, VU0453595 produced dose-related increases in high frequency gamma power, a correlate of arousal and cognition enhancement, without altering duration of time across all sleep/wake stages. Effects of VU0453595 in NHPs were observed within a dose range that did not induce cholinergic-mediated adverse effects. In contrast, donepezil and xanomeline increased time awake in rodents and engendered dose-limiting adverse effects in NHPs. Finally, VU0453595 attenuated age-related decreases in REM sleep duration in aged wildtype mice. Development of M(1) PAMs represents a viable strategy for attenuating age-related and dementia-related pathological disturbances of sleep and arousal.

In this manuscript, we report a series of chiral 6-azaspiro[2.5]octanes and related spirocycles as highly potent and selective antagonists of the muscarinic acetylcholine receptor subtype 4 (mAChR4). Chiral separation and subsequent X-ray crystallographic analysis of early generation analogs revealed the R enantiomer to possess excellent human and rat M4 potency, and further structure-activity relationship (SAR) studies on this chiral scaffold led to the discovery of VU6015241 (compound 19). Compound 19 is characterized by high M4 potency and selectivity across multiple species, excellent aqueous solubility, and moderate brain exposure in rodents after intraperitoneal administration.

PURPOSE: To test the psychometric properties of the European Organisation for Research and Treatment of Cancer (EORTC) Quality of Life Questionnaire–Chemotherapy-Induced Peripheral Neuropathy (QLQ-CIPN20) using Rasch-based methods. METHODS: A secondary data analysis was performed using pooled QLQ-CIPN20 data from patients (N = 1008) who had participated in any of four multi-site chemotherapy-induced peripheral neuropathy (CIPN) treatment and prevention trials. QLQ-CIPN20 responses were evaluated using a polytomous Rasch partial credit model. Data were assessed for person-item fit using the chi-square statistic, item scaling based on response proportions, threshold ordering using item characteristic curves and logit threshold locations, differential item response (DIF) (i.e., response bias) using likelihood ratio tests, and unidimensionality using cluster analysis. RESULTS: A statistically significant chi-square test indicated poor fit of the observed to the expected responses. More than 70% of the respondents reported a complete absence of six symptoms, reflecting significant floor effects and poor item scaling. Disordered/non-ordinal or narrow response thresholds were found for 11 of the 20 items. Item responses were significantly different by gender (p < 0.0001) and chemotherapy type (p < 0.0001). Cluster analysis findings suggest that the QLQ-CIPN20 is a unidimensional scale due to the absence of item clusters. CONCLUSIONS: Rasch model testing revealed psychometric weaknesses that could be addressed by revising the QLQ-CIPN20’s problematic items and response options. Alternatively, perhaps the new gold standard CIPN measurement approach in future intervention trials should involve use of only the best items, which would also allow comparisons across previous trials that utilized the QLQ-CIPN20.

This chapter examines the dual system of family law in the USA. It observes that the USA has a set of laws that regulates more affluent families and an entirely distinct set of laws that regulates poor families. Moreover, the family law for the poor is uniquely punitive. The chapter offers that the dual system of family law, and the brutal nature of family law for the poor, can be explained in terms of the moral construction of poverty—the idea that poverty is a result of an individual’s shortcomings. The chapter proposes that the moral construction of poverty offers a unique framework through which to view and critique the family law for the poor. It demonstrates the utility of the framework through an analysis of the Court’s 1970 decision in Dandridge v. Williams, upholding the constitutionality of family cap policies that restrict the size of the grants that welfare beneficiaries receive to support their families.

The pharmacology of the M5 muscarinic acetylcholine receptor (mAChR) is the least understood of the five mAChR subtypes due to a historic lack of selective small molecule tools. To address this shortcoming, we have continued the optimization effort around the prototypical M5 positive allosteric modulator (PAM) ML380 and have discovered and optimized a new series of M5 PAMs based on a chiral N-(indanyl)piperidine amide core with robust SAR, human and rat M5 PAM EC50 values

Although selective activation of the M1 muscarinic acetylcholine receptor (mAChR) subtype has been shown to improve cognitive function in animal models of neuropsychiatric disorders, recent evidence suggests that enhancing M4 mAChR function can also improve memory performance. Positive allosteric modulators (PAMs) targeting the M4 mAChR subtype have shown therapeutic potential for the treatment of multiple symptoms observed in schizophrenia, including positive and cognitive symptoms when assessed in acute preclinical dosing paradigms. Since the cholinergic system has been implicated in multiple stages of learning and memory, we evaluated the effects of repeated dosing with the highly selective M4 PAM VU0467154 on either acquisition and/or consolidation of learning and memory when dosed alone or after pharmacologic challenge with the N-methyl-d-aspartate subtype of glutamate receptors (NMDAR) antagonist MK-801. MK-801 challenge represents a well-documented preclinical model of NMDAR hypofunction that is thought to underlie some of the positive and cognitive symptoms observed in schizophrenia. In wildtype mice, 10-day, once-daily dosing of VU0467154 either prior to, or immediately after daily testing enhanced the rate of learning in a touchscreen visual pairwise discrimination task; these effects were absent in M4 mAChR knockout mice. Following a similar 10-day, once-daily dosing regimen of VU0467154, we also observed 1) improved acquisition of memory in a cue-mediated conditioned freezing paradigm, 2) attenuation of MK-801-induced disruptions in the acquisition of memory in a context-mediated conditioned freezing paradigm and 3) reversal of MK-801-induced hyperlocomotion. Comparable efficacy and plasma and brain concentrations of VU0467154 were observed after repeated dosing as those previously reported with an acute, single dose administration of this M4 PAM. Together, these studies are the first to demonstrate that cognitive enhancing and antipsychotic-like activity are not subject to the development of tolerance following repeated dosing with a selective M4 PAM in mice and further suggest that activation of M4 mAChRs may modulate both acquisition and consolidation of memory functions.

This Letter details our efforts to develop novel tricyclic M4 PAM scaffolds with improved pharmacological properties. This endeavor involved a "tie-back" strategy to replace the 3-amino-4,6-dimethylthieno[2,3-b]pyridine-2-carboxamide core which lead to the discovery of two novel tricyclic cores: a 7,9-dimethylpyrido[3',2':4,5]thieno[3,2-d]pyrimidine core and 2,4-dimethylthieno[2,3-b:5,4-c']dipyridine core. Both tricyclic cores displayed low nanomolar potency against the human M4 receptor.

Purpose Current melphalan-based regimens for intravitreal chemotherapy for retinoblastoma vitreous seeds are effective but toxic to the retina. Thus, alternative agents are needed. Based on the known biology of histone deacetylases (HDACs) in the retinoblastoma pathway, we systematically studied whether the HDAC inhibitor belinostat is a viable, molecularly targeted alternative agent for intravitreal delivery that might provide comparable efficacy, without toxicity. Methods In vivo pharmacokinetic experiments in rabbits and in vitro cytotoxicity experiments were performed to determine the 90% inhibitory concentration (IC90). Functional toxicity by electroretinography and structural toxicity by optical coherence tomography (OCT), OCT angiography, and histopathology were evaluated in rabbits following three injections of belinostat 350 µg (2× IC90) or 700 µg (4× IC90), compared with melphalan 12.5 µg (rabbit equivalent of the human dose). The relative efficacy of intravitreal belinostat versus melphalan to treat WERI-Rb1 human cell xenografts in rabbit eyes was directly quantified. RNA sequencing was used to assess belinostat-induced changes in RB cell gene expression. Results The maximum nontoxic dose of belinostat was 350 µg, which caused no reductions in electroretinography parameters, retinal microvascular loss on OCT angiography, or retinal degeneration. Melphalan caused severe retinal structural and functional toxicity. Belinostat 350 µg (equivalent to 700 µg in the larger human eye) was equally effective at eradicating vitreous seeds in the rabbit xenograft model compared with melphalan (95.5% reduction for belinostat, P < 0.001; 89.4% reduction for melphalan, P < 0.001; belinostat vs. melphalan, P = 0.10). Even 700 µg belinostat (equivalent to 1400 µg in humans) caused only minimal toxicity. Widespread changes in gene expression resulted. Conclusions Molecularly targeted inhibition of HDACs with intravitreal belinostat was equally effective as standard-of-care melphalan but without retinal toxicity. Belinostat may therefore be an attractive agent to pursue clinically for intravitreal treatment of retinoblastoma.

N-Alkyl substituted chlorotetrazines were coupled with various boronic acids under Suzuki conditions in high yield at room temperature, giving a mild and straightforward synthetic route toward diverse unsymmetrical 1,2,4,5-tetrazines, a rare heteroarene. This chemistry not only expands the known substrate scope of tetrazine cross-coupling reactions but also allows for the synthesis of novel, tetrazine-containing biologically active molecules with improved DMPK properties.

This Letter details the discovery and subsequent optimization of a novel M4 PAM scaffold based on an 6-fluoro-4-(piperidin-1-yl)quinoline-3-carbonitrile core, which represents a distinct departure from the classical M4 PAM chemotypes. Optimized compounds in this series demonstrated improved M4 PAM potency on both human and rat M4 (4 to 5-fold relative to HTS hit), and displayed attractive physicochemical and DMPK profiles, including good CNS penetration (rat brain:plasma Kp=5.3, Kp,uu=2.4; MDCK-MDR1 (P-gp) ER=1.1).

This Letter describes the synthesis and structure activity relationship (SAR) studies of structurally novel M(4) antagonists, based on a 3-(4-aryl/heteroarylsulfonyl)piperazin-1-yl)-6-(piperidin-1-yl)pyridazine core, identified from a high-throughput screening campaign. A multidimensional optimization effort enhanced potency at human M(4) (hM(4) IC(50)s < 200 nM), with only moderate species differences noted, and with enantioselective inhibition. Moreover, CNS penetration proved attractive for this series (rat brain:plasma K(p) = 2.1, K(p,uu) = 1.1). Despite the absence of the prototypical mAChR antagonist basic or quaternary amine moiety, this series displayed pan-muscarinic antagonist activity across M(1-5) (with 9- to 16-fold functional selectivity at best). This series further expands the chemical diversity of mAChR antagonists.