Impact of Indole Inclusion in the Design of Multi-Tactical Metal-Binding Tetra-Aza Macrocycles that Target the Molecular Features of Neurodegeneration.

Numerous small molecules have been studied for their ability to counteract oxidative stress, a key contributor to neurodegenerative diseases such as Alzheimer's. Despite these efforts, the pharmacological properties and structure-activity relationships of these compounds remain insufficiently understood, yet they are critical in evaluating a drug molecule's therapeutic potential. A modified tetra-aza macrocycle has demonstrated strong antioxidant activity through various mechanisms; however, its limited permeability presents challenges for advanced formulation studies. To enhance permeability while preserving the beneficial reactivity of the parent molecule, two synthetic modifications involving indole functionality were explored and compared to modifications using methyl groups alone. New synthetic strategies were developed to produce the indole-containing molecules, which were characterized by 1D/2D NMR techniques. Isoelectric points, metal binding, and radical scavenging activity were determined to validate that the reactivity of the parent molecules was retained. The permeability of all molecules explored was improved. Protection against oxidative stress through activation of the Nrf2 pathway was demonstrated for molecules containing indoles in cellular models by measuring ROS levels upon treatment and mRNA levels of HO-1 and Nrf2. In contrast, no protection or Nrf2 activation was observed with the methylation of the O- or N atom. These results suggest that while alkylation improves permeability overall, concomitant antioxidant protection and positive permeability are achieved with the indole congeners alone.

Economic Burden of Foodborne Illnesses Acquired in the United States.

Foodborne disease burden estimates inform public health priorities and can help the public understand disease impact. This article provides new estimates of the cost of U.S. foodborne illness. Our research updated disease modeling underlying these cost estimates with a focus on enhancing chronic sequelae modeling and enhancing uncertainty modeling. Our cost estimates were based on U.S. Centers for Disease Control and Prevention estimates of the numbers of foodborne illnesses, hospitalizations, and deaths caused by 31 known foodborne pathogens and unspecified foodborne agents. We augmented these estimates of illnesses, hospitalizations, and deaths with more detailed modeling of health outcomes, including chronic sequelae. For health outcomes, we relied on U.S. data and research where possible, supplemented by the use of non-U.S. research where necessary and scientifically appropriate. Cost estimates were developed from large insurance or hospital charge databases, public data sources, and existing literature and were adjusted to 2023 dollars. We estimated the cost of foodborne illness in the United States circa 2023 to be $75 billion. Deaths accounted for 56% and chronic outcomes for 31% of the mean cost. The costliest pathogen was nontyphoidal Salmonella at $17.1 billion followed by Campylobacter at $11.3 billion. Toxoplasma ($5.7 billion) and Listeria ($4 billion) followed due primarily to deaths and chronic outcomes from pregnancy-associated cases. Per-case cost ranged from $196 for Bacillus cereus to $4.6 million for Vibrio vulnificus. Unspecified agents accounted for 38% of the total cost of foodborne illness, but these illnesses were generally mild (per-case cost $781). These cost estimates can help inform food safety priorities. Our pathogen-specific per-case cost estimates can also help inform benefit-cost analysis required for new federal food safety regulations.

Single Cell Expression Analysis of Ductal Carcinoma in Situ Identifies Complex Genotypic-Phenotypic Relationships Altering Epithelial Composition.

To identify mechanisms underlying the growth of ductal carcinoma in situ (DCIS) and properties that lead to progression to invasive cancer, we performed single-cell RNA-sequencing (scRNA-seq) on DCIS lesions and matched synchronous normal breast tissue. Using inferred copy number variations (CNV), we identified neoplastic epithelial cells from the clinical specimens which contained a mixture of DCIS and normal ducts. Phylogenetic analysis based on the CNVs demonstrated intratumoral clonal heterogeneity was associated with significant gene expression differences. We also classified epithelial cells into mammary cell states and found that individual genetic clones contained a mixture of cell states suggesting an ongoing pattern of differentiation after neoplastic transformation. Cell state proportions were significantly different based on estrogen receptor (ER) expression with ER-DCIS more closely resembling the distribution in the normal breast, particularly with respect to cells with basal characteristics. Using deconvolution from bulk RNA-seq in archival DCIS specimens, we show that specific alterations in cell state proportions are associated with progression to invasive cancer. Loss of an intact basement membrane (BM) is the functional definition of invasive breast cancer (IBC) and scRNA-seq data demonstrated that ongoing transcription of key BM genes occurs in specific subsets of epithelial cell states. Examining BM in archival microinvasive breast cancers and an in vitro model of invasion, we found that passive loss of BM gene expression due to cell state proportion alterations is associated with loss of the structural integrity of the duct leading to an invasive phenotype. Our analyses provide detailed insight into DCIS biology. SIGNIFICANCE: Single cell analysis reveals that preinvasive breast cancer is comprised of multiple genetic clones and there is substantial phenotypic diversity both within and between these clones. Ductal carcinoma in situ (DCIS) of the breast is a non-invasive condition commonly identified through mammographic screening. A primary diagnosis of DCIS carries little mortality risk on its own, but its presence is a risk factor for subsequent clonally related invasive breast cancer (IBC) (1-5).

Electronic angle focusing for neutron time-of-flight powder diffractometers.

A neutron time-of-flight (TOF) powder diffractometer with a continuous wide-angle array of detectors can be electronically focused to make a single pseudo-constant wavelength diffraction pattern, thus facilitating angle-dependent intensity corrections. The resulting powder diffraction peak profiles are affected by the neutron source emission profile and resemble the function currently used for TOF diffraction.

Optimization of testing protocols to screen for COVID-19: a multi-objective model.

In this paper we develop a new multi-objective simulated annealing (MOSA) algorithm to generate optimal testing protocols for infectious diseases, using the COVID-19 pandemic as our context. A SEIR (susceptible-exposed-infected-recovered) epidemiological model is embedded as the computational platform for our MOSA algorithm to optimize testing protocols for screening across three joint objectives: minimum cost of test materials, minimum total infections over the testing horizon, and minimum number of false negatives over the horizon. We demonstrate the application of this optimization tool to recommend screening protocols for K-12 school districts in the U.S. State of North Carolina. Our approach is scalable by population coverage and can be employed at the level of individual school districts or regional collections of districts, individual schools or collections of schools across a district, business sites, or nursing homes, among other congregate settings where individuals may be screened prior to gaining entry to the site. The algorithm can be solved two ways, generating either independent optimal protocols across individual testing locations, or a common protocol covering all locations in the collection of testing sites. Our findings can be used to inform policy decisions to guide the development of effective testing strategies for controlling the spread of COVID-19 or other pandemic diseases in a wide range of congregate settings across various geographic regions.

Prevalence and incidence of uveitis in patients with spondyloarthritis: the impact of the biologics era. Data from International ASAS-COMOSPA Study.

OBJECTIVES: Uveitis is a common extra-musculoskeletal manifestation in Spondyloarthritis (SpA). The aim of this study was to analyze the prevalence of uveitis in SpA patients, its association with geographical areas and to determine whether its incidence was different before and after the biological era. METHODS: ASAS-COMOSPA is a retrospective study that includes patients fulfilling ASAS SpA classification criteria from 22 countries. The overall prevalence of uveitis was calculated, and factors associated with the onset of a first episode of uveitis were evaluated using a Cox regression. A Log-Rank test was performed to compare the new onset of uveitis in the no biological era (SpA onset before 2000) vs biological era (SpA onset after 2000). RESULTS: 3984 patients were included. The likelihood of presenting a first uveitis episode increased over time, from a prevalence of 10.5% (95%CI 9.5%-11.4%) at the time of the SpA diagnosis to 46.6% (41.6%-51.5%) after 30 years since the SpA diagnosis. HLA-B27 positivity, family history of uveitis, peripheral enthesitis and IBD were associated with higher risk of uveitis. Patients with SpA disease onset after year 2000 showed a lower prevalence of uveitis compared with disease onset before year 2000 (8.2% vs 25.5%, p< 0.01), as well as a lower incidence (2.8 per 100 PY vs 6.1 per 100 PY, respectively). CONCLUSION: In our study the risk of having suffered from at least one episode of uveitis ranged from 10% at the time of the diagnosis of axSpA to 47% after 30 years of disease duration. Patients with disease onset after biologic therapy introduction showed a significantly lower prevalence and incidence of first episodes of uveitis.

Functional germline variants in DNA damage repair pathways are associated with altered survival in adults with glioma treated with temozolomide.

Background: Temozolomide (TMZ) treatment has demonstrated, but variable, impact on glioma prognosis. This study examines associations of survival with DNA repair gene germline polymorphisms among glioma patients who did and did not have TMZ treatment. Identifying genetic markers which sensitize tumor cells to TMZ could personalize therapy and improve outcomes. Methods: We evaluated TMZ-related survival associations of pathogenic germline SNPs and genetically predicted transcript levels within 34 DNA repair genes among 1504 glioma patients from the UCSF Adult Glioma Study and Mayo Clinic whose diagnoses spanned pre- and post-TMZ eras within the major known glioma prognostic molecular subtypes. Results: Among those who received TMZ, 5 SNPs were associated with overall survival, but not in those who did not receive TMZ. Only rs2308321-G, in MGMT , was associated with decreased survival (HR=1.21, p=0.019) for all glioma subtypes. Rs73191162-T (near UNG ), rs13076508-C (near PARP3 ), rs7840433-A (near NEIL2 ), and rs3130618-A (near MSH5 ) were only associated with survival and TMZ treatment for certain subtypes, suggesting subtype-specific germline chemo-sensitization. Genetically predicted elevated compared to normal brain expression of PNKP was associated with dramatically worse survival for TMZ-treated patients with IDH -mutant and 1p/19q non-codeleted gliomas (p=0.015). Similarly, NEIL2 and TDG expressions were associated with altered TMZ-related survival only among certain subtypes. Conclusions: Functional germline alterations within DNA repair genes were associated with TMZ sensitivity, measured by overall survival, among adults with glioma, these variants should be evaluated in prospective analyses and functional studies. Key points: We observed SNPs associated with glioma survival, specific to cases receiving TMZ An MGMT variant may reduce glioma survival indirectly through myelosuppression Decreased genetic PNKP expression in the brain may sensitize cells to TMZ. Importance of the study: The introduction of temozolomide (TMZ) as a part of standard-of-care in the treatment of gliomas marked the last notable increase in patient survival. However, the effectiveness of TMZ is not universal, and can result in serious complications. The mechanism of action behind the drug is the introduction of damaging methyl groups across the tumor genome and leveraging of DNA damage repair (DDR) mechanisms to signal programmed cell death. Previous literature has identified that defects in DDR mechanisms can alter TMZ sensitivity. Using a unique dataset that spans the pre- and post-TMZ eras, we demonstrate that germline variation in DDR-related genes may have significant impact on overall survival for patients treated with TMZ, with no effects observed in the pre-TMZ era. This suggests that germline variants in these DDR genes could be used to personalize TMZ therapy to improve patient survival.

Effect of chiropractic care on low back pain for active-duty military members: Mediation through biopsychosocial factors.

This study evaluates biopsychosocial factors as mediators of the effect of chiropractic care on low back pain (LBP) intensity and interference for active-duty military members. Data from a multi-site, pragmatic clinical trial comparing six weeks of chiropractic care plus usual medical care to usual medical care alone for 750 US active-duty military members with LBP were analyzed using natural-effect, multiple-mediator modeling. Mediation of the adjusted mean effect difference on 12-week outcomes of PROMIS-29 pain interference and intensity by 6-week mediators of other PROMIS-29 physical, mental, and social health subdomains was evaluated. The effect difference on pain interference occurring through PROMIS-29 biopsychosocial factors (natural indirect effect = -1.59, 95% CI = -2.28 to -0.88) was 56% (95% CI = 35 to 96) of the total effect (-2.82, 95% CI = -3.98 to -1.53). The difference in effect on pain intensity occurring through biopsychosocial factors was smaller (natural indirect effect = -0.32, 95% CI = -0.50 to -0.18), equaling 26% (95% CI = 15 to 42) of the total effect (-1.23, 95% CI = -1.52 to -0.88). When considered individually, all physical, mental, and social health factors appeared to mediate the effect difference on pain interference and pain intensity with mental health factors having smaller effect estimates. In contrast with effects on pain interference, much of the effect of adding chiropractic care to usual medical care for US military members on pain intensity did not appear to occur through the PROMIS-29 biopsychosocial factors. Physical and social factors appear to be important intermediate measures for patients receiving chiropractic care for low back pain in military settings. Further study is needed to determine if the effect of chiropractic care on pain intensity for active-duty military occurs through other unmeasured factors, such as patient beliefs, or if the effect occurs directly.

Emulating the Delivery of Sawtooth Proton Arc Therapy Plans on a Cyclotron-Based Proton Beam Therapy System.

Purpose: To evaluate and compare the deliverability of 'sawtooth' proton arc therapy (PAT) plans relative to static intensity modulated proton therapy (IMPT) at a cyclotron-based clinical facility. Methods: The delivery of single and dual arc Sawtooth PAT plans for an abdominal CT phantom and multiple clinical cases of brain, head and neck (H&N) and base of skull (BoS) targets was emulated under the step-and-shoot and continuous PAT delivery regimes and compared to that of a corresponding static IMPT plan. Results: Continuous PAT delivery increased the time associated with beam delivery and gantry movement in single/dual PAT plans by 4.86/7.34 min (brain), 7.51/12.40 min (BoS) and 6.59/10.57 min (H&N) on average relative to static IMPT. Step-and-shoot PAT increased this delivery time further by 4.79 min on average as the delivery was limited by gantry motion. Conclusions: The emulator can approximately model clinical sawtooth PAT delivery but requires experimental validation. No clear benefit was observed regarding beam-on time for sawtooth PAT relative to static IMPT.

Mesoscale properties of biomolecular condensates emerging from protein chain dynamics.

Biomolecular condensates form by phase separation of biological polymers. The cellular functions of the resulting membraneless organelles are closely linked to their physical properties over a wide range of length- and timescales: From the nanosecond dynamics of individual molecules and their interactions, to the microsecond translational diffusion of molecules in the condensates, to their viscoelastic properties at the mesoscopic scale. However, it has remained unclear how to quantitatively link these properties across scales. Here we address this question by combining single-molecule fluorescence, correlation spectroscopy, microrheology, and large-scale molecular dynamics simulations on different condensates that are formed by complex coacervation and span about two orders of magnitude in viscosity and their dynamics at the molecular scale. Remarkably, we find that the absolute timescale of protein chain dynamics in the dense phases can be quantitatively and accurately related to translational diffusion and condensate viscosities by Rouse theory of polymer solutions including entanglement. The simulations indicate that the observed wide range of dynamics arises from different contact lifetimes between amino acid residues, which in the mean-field description of the polymer model cause differences in the friction acting on the chains. These results suggest that remarkably simple physical principles can relate the mesoscale properties of biomolecular condensates to their dynamics at the nanoscale.

ß-synuclein regulates the phase transitions and amyloid conversion of α-synuclein.

Parkinson's disease (PD) and Dementia with Lewy Bodies (DLB) are neurodegenerative disorders characterized by the accumulation of α-synuclein aggregates. α-synuclein forms droplets via liquid-liquid phase separation (LLPS), followed by liquid-solid phase separation (LSPS) to form amyloids, how this process is physiologically-regulated remains unclear. ß-synuclein colocalizes with α-synuclein in presynaptic terminals. Here, we report that ß-synuclein partitions into α-synuclein condensates promotes the LLPS, and slows down LSPS of α-synuclein, while disease-associated ß-synuclein mutations lose these capacities. Exogenous ß-synuclein improves the movement defects and prolongs the lifespan of an α-synuclein-expressing NL5901 Caenorhabditis elegans strain, while disease-associated ß-synuclein mutants aggravate the symptoms. Decapeptides targeted at the α-/ß-synuclein interaction sites are rationally designed, which suppress the LSPS of α-synuclein, rescue the movement defects, and prolong the lifespan of C. elegans NL5901. Together, we unveil a Yin-Yang balance between α- and ß-synuclein underlying the normal and disease states of PD and DLB with therapeutical potentials.

Subsurface Microbial Colonization at Mineral-Filled Veins in 2-Billion-Year-Old Mafic Rock from the Bushveld Igneous Complex, South Africa.

Recent advances in subsurface microbiology have demonstrated the habitability of multi-million-year-old igneous rocks, despite the scarce energy supply from rock-water interactions. Given the minimal evolution coupled with exceedingly slow metabolic rates in subsurface ecosystems, spatiotemporally stable igneous rocks can sustain microbes over geological time scales. This study investigated a 2-billion-year-old mafic rock in the Bushveld Igneous Complex, South Africa, where ultradeep drilling is being executed by the International Continental Scientific Drilling Program (ICDP). New procedures were successfully developed to simultaneously detect indigenous and contaminant microbial cells in a drill core sample. Precision rock sectioning coupled with infrared, fluorescence, and electron microscopy imaging of the rock section with submicron resolution revealed microbial colonization in veins filled with clay minerals. The entry and exit of microbial cells in the veins are severely limited by tight packing with clay minerals, the formation of which supplies energy sources for long-term habitability. Further microbiological characterization of drilled rock cores from the Bushveld Igneous Complex will expand the understanding of microbial evolution in deep igneous rocks over 2 billion years.

Increased TSPO, COX-1, and COX-2 Uptake in Bone Marrow Within a Falx Cerebri Ossification.

ABSTRACT: Ossification is uncommon, generally asymptomatic, and often incidentally identified in imaging studies. We report on a 54-year-old man who participated as a healthy volunteer in a clinical trial using PET imaging to investigate neuroinflammation. An incidental ossified lesion in the anterior falx cerebri was revealed by MRI. CT scan showed a small hypodense center in the lesion, probably corresponding to bone marrow. The PET scans using 18F-SF51, 11C-PS13, and 11C-MC1 showed increased uptake within this lesion, which was probably related to bone marrow activity within the ossification.

Contrasting Genomic Diversity and Inbreeding Levels Among Two Closely Related Falcon Species With Overlapping Geographic Distributions.

Genomic resources are valuable to examine historical demographic patterns and their effects to better inform management and conservation of threatened species. We evaluated population trends and genome-wide variation in the near-threatened Orange-breasted Falcon (Falco deiroleucus) and its more common sister species, the Bat Falcon (F. rufigularis), to explore how the two species differ in genomic diversity as influenced by their contrasting long-term demographic histories. We generated and aligned whole genome resequencing data for 12 Orange-breasted Falcons and 9 Bat Falcons to an annotated Gyrfalcon (F. rusticolus) reference genome that retained approximately 22.4 million biallelic autosomal SNPs (chromosomes 1-22). Our analyses indicated much lower genomic diversity in Orange-breasted Falcons compared to Bat Falcons. All sampled Orange-breasted Falcons were significantly more inbred than the sampled Bat Falcons, with values similar to those observed in island-mainland species comparisons. The distribution of runs of homozygosity showed variation suggesting long-term low population size and the possibility of bottlenecks in Orange-breasted Falcons contrasting with consistently larger populations in Bat Falcons. Analysis of genetic load suggests that Orange-breasted Falcons are less likely to experience inbreeding depression than Bat Falcons due to reduced inbreeding load but are at elevated risk from fixation of deleterious gene variants and perhaps a reduced adaptive potential. These genomic analyses highlight differences in the historical demography of two closely related species that have influenced their current genomic diversity and should result in differing strategies for their continued conservation.

Bronchial cell epigenetic aging in a human experimental study of short-term diesel and ozone exposures.

Blood-based, observational, and cross-sectional epidemiological studies suggest that air pollutant exposures alter biological aging. In a single-blinded randomized crossover human experiment of 17 volunteers, we examined the effect of randomized 2-h controlled air pollution exposures on respiratory tissue epigenetic aging. Bronchial epithelial cell DNA methylation 24 h post-exposure was measured using the HumanMethylation450K BeadChip, and there was a minimum 2-week washout period between exposures. All 17 volunteers were exposed to ozone, but only 13 were exposed to diesel exhaust. Horvath DNAmAge [Pearson coefficient (r) = 0.64; median absolute error (MAE) = 2.7 years], GrimAge (r = 0.81; MAE = 13 years), and DNAm Telomere Length (DNAmTL) (r = -0.65) were strongly correlated with chronological age in this tissue. Compared to clean air, ozone exposure was associated with longer DNAmTL (median difference 0.11 kb, Fisher's exact P-value = .036). This randomized trial suggests a weak relationship of ozone exposure with DNAmTL in target respiratory cells. Still, causal relationships with long-term exposures need to be evaluated.

Early detection of ovarian cancer using cell-free DNA fragmentomes and protein biomarkers.

Ovarian cancer is a leading cause of death for women worldwide in part due to ineffective screening methods. In this study, we used whole-genome cell-free DNA (cfDNA) fragmentome and protein biomarker (CA-125 and HE4) analyses to evaluate 591 women with ovarian cancer, benign adnexal masses, or without ovarian lesions. Using a machine learning model with the combined features, we detected ovarian cancer with specificity >99% and sensitivity of 72%, 69%, 87%, and 100% for stages I-IV, respectively. At the same specificity, CA-125 alone detected 34%, 62%, 63%, and 100% of ovarian cancers for stages I-IV. Our approach differentiated benign masses from ovarian cancers with high accuracy (AUC=0.88, 95% CI=0.83-0.92). These results were validated in an independent population. These findings show that integrated cfDNA fragmentome and protein analyses detect ovarian cancers with high performance, enabling a new accessible approach for noninvasive ovarian cancer screening and diagnostic evaluation.

Neonatal Clinical Pharmacology.

Similar to other populations, care of neonates strongly relies on robust guidance on neonatal pharmacotherapy, covering an age-appropriate drug formulation, an individualized dose, and information on its efficacy and safety for a specific indication in this population, preferably weighted to alternative approaches [...].

Experiences in Coping with Stress-A Qualitative Study of Family Caregivers of Children with Medical Complexity.

OBJECTIVE: To better understand the strategies family caregivers of children with medical complexity (CMC) utilize to deal with the stress and challenges associated with caregiving. METHODS: We conducted a cross-sectional qualitative study among family caregivers of CMC receiving medical care at a children's hospital in Western Pennsylvania. Participants completed in-depth, semi-structured interviews focused on how CMC family caregivers approach and manage caregiving-related challenges and stress. Using constant comparative methodology, we inductively analyzed deidentified transcripts for emergent themes. RESULTS: We interviewed 19 participants (89.4% female) with a mean age of 43 years (range 32-54 years). The mean age of the participants' children was 10.8 years (range 1-20 years). Twelve participants' children identified as white and four identified as Black. Three central themes regarding CMC caregivers' stress-coping strategies emerged: (1) maintaining a positive mindset, (2) developing and relying on interpersonal support networks, and (3) making time for self-preservation. All three themes were universally reported (n = 19/19) by our participants. The most common subthemes for each theme, respectively, focused on staying hopeful and celebrating moments of joy; cultivating supportive relationships with family, friends, and fellow CMC family caregivers; and finding pleasure in "little things" (e.g., everyday activities and hobbies). CONCLUSION: Family caregivers of CMC utilize a multi-faceted approach to cope with the stress and challenges routinely encountered in caring for CMC. This study's findings could be used to inform future clinical efforts and research directions aiming to improve clinicians' ability to support CMC caregivers' well-being.

Trends in hip fracture rates in US male veterans.

Osteoporotic fracture has been understudied in men. In US male veterans aged 50 years and older between 2002 and 2019, hip fracture incidence increased between 2006 and 2019, fewer than 6% of men underwent DXA, and fewer than 0.5% of men were treated. Investigation of low screening and treatment rates is warranted. PURPOSE: In the United States, the annual incidence of osteoporotic hip fracture is estimated to be 250,000 to 300,000; the one-year mortality in some studies has been as high as 32%. Reports that hip fracture rates in US women 65 years and older may no longer be declining led to this investigation of hip fracture in men, a less studied population. We assessed the trends in the incidence of hip fracture in US male veterans 50 years and older of age as well as the rates of diagnosis and treatment in such men. METHODS: We assessed the recent trends of hip fracture incidence in a nation-wide male veteran population 50 years and older of age. Using data from the US Veterans Affairs Informatics and Computing Infrastructure (VINCI) 2002-2019, we calculated the annual age-standardized hip fracture incidence. Secondary objectives included evaluating the annual proportion of hip fracture patients who underwent dual-energy X-ray absorptiometry (DXA) before or after the fracture and/or received osteoporosis medication after the hip fracture over the study period. RESULTS: Hip fracture incidence increased in male veterans from 2006 to 2019. Fewer than 6% of men underwent a DXA scan and fewer than 0.5% received osteoporosis medications up to two years after a hip fracture. CONCLUSIONS: Despite available screening methods such as DXAs and medications for primary and secondary prevention of osteoporotic fractures, hip fracture incidence is not decreasing in older male veterans. Our study highlights a need for closer attention to fracture risk in men.