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Abstract Prognostic analysis for early-stage (stage I/II) melanomas is of paramount importance for customized surveillance and treatment plans. Since immune checkpoint inhibitors have recently been approved for stage IIB and IIC melanomas, prognostic tools to identify patients at high risk of recurrence have become even more critical. This study aims to assess the effectiveness of machine-learning algorithms in predicting melanoma recurrence using clinical and histopathologic features from Electronic Health Records (EHRs). We collected 1720 early-stage melanomas: 1172 from the Mass General Brigham healthcare system (MGB) and 548 from the Dana-Farber Cancer Institute (DFCI). We extracted 36 clinicopathologic features and used them to predict the recurrence risk with supervised machine-learning algorithms. Models were evaluated internally and externally: (1) five-fold cross-validation of the MGB cohort; (2) the MGB cohort for training and the DFCI cohort for testing independently. In the internal and external validations, respectively, we achieved a recurrence classification performance of AUC: 0.845 and 0.812, and a time-to-event prediction performance of time-dependent AUC: 0.853 and 0.820. Breslow tumor thickness and mitotic rate were identified as the most predictive features. Our results suggest that machine-learning algorithms can extract predictive signals from clinicopathologic features for early-stage melanoma recurrence prediction, which will enable the identification of patients that may benefit from adjuvant immunotherapy.

Abstract Background Patients with language barriers encounter healthcare disparities, which may be alleviated by leveraging interpreter skills to reduce cultural, language, and literacy barriers through improved bidirectional communication. Evidence supports the use of in-person interpreters, especially for interactions involving patients with complex care needs. Unfortunately, due to interpreter shortages and clinician underuse of interpreters, patients with language barriers frequently do not get the language services they need or are entitled to. Health information technologies (HIT), including artificial intelligence (AI), have the potential to streamline processes, prompt clinicians to utilize in-person interpreters, and support prioritization. Methods From May 1, 2023, to June 21, 2024, a single-center stepped wedge cluster randomized trial will be conducted within 35 units of Saint Marys Hospital & Methodist Hospital at Mayo Clinic in Rochester, Minnesota. The units include medical, surgical, trauma, and mixed ICUs and hospital floors that admit acute medical and surgical care patients as well as the emergency department (ED). The transitions between study phases will be initiated at 60-day intervals resulting in a 12-month study period. Units in the control group will receive standard care and rely on clinician initiative to request interpreter services. In the intervention group, the study team will generate a daily list of adult inpatients with language barriers, order the list based on their complexity scores (from highest to lowest), and share it with interpreter services, who will send a secure chat message to the bedside nurse. This engagement will be triggered by a predictive machine-learning algorithm based on a palliative care score, supplemented by other predictors of complexity including length of stay and level of care as well as procedures, events, and clinical notes. Discussion This pragmatic clinical trial approach will integrate a predictive machine-learning algorithm into a workflow process and evaluate the effectiveness of the intervention. We will compare the use of in-person interpreters and time to first interpreter use between the control and intervention groups. Trial registration NCT05860777. May 16, 2023.

Abstract Clustered regularly interspaced short palindromic repeats (CRISPR) technology has revolutionized creating targeted genetic variation in crops. Although CRISPR enzymes have been reported to have high sequence‐specificity, careful design of the editing reagents can also reduce unintended edits at highly homologous sites. This work details the first large‐scale study of the heritability of on‐target edits and the rate of edits at off‐target sites in soybean (Glycine max), assaying ~700 T1 plants each resulting from transformation with LbCas12a constructs containing CRISPR RNAs (crRNAs) predicted to be either “unique” with no off‐target sites or “promiscuous” with >10 potential off‐targets in the soybean genome. Around 80% of the on‐target edits observed in T0 plants were inherited in the T1 generation, and ~49% of the total observed on‐target edits in T1 were not observed at T0, indicating continued activity of LbCas12a throughout the life cycle of the plant. In planta editing at off‐target sites was observed for the Promiscuous but not the Unique crRNA. Examination of the edited off‐target sites revealed that LbCas12a was highly tolerant to mismatches between the crRNA and target site in bases 21–23 relative to the start of the protospacer, but even a single mismatch in the first 20 nt drastically reduced the editing rate. In addition, edits at off‐target sites have lower inheritance rates than on‐target edits, suggesting that they occur later in the plant's lifecycle. Plants with a desired on‐target edit and no off‐target edits could be identified in the T1 generation for 100% of the T0 plants edited with the Unique crRNA compared with the 65% of T0 plants edited with the Promiscuous crRNA. This confirms that proper crRNA selection can reduce or eliminate off‐target editing. Even when potential off‐target sites are predicted, plants containing only the intended edits can still be identified and propagated.

Despite the acknowledged impact of early science achievement on future success, there is a noticeable gap in research focused on understanding the dynamic longitudinal patterns of children attaining science learning milestones in their early years, as well as few investigations of potential factors that may mitigate gender, racial/ethnic, and socio-economic disparities. This study analyzed nationally representative data from the Early Childhood Longitudinal Study of Kindergarten Cohort 2011 (ECLS-K: 2011), involving 18,174 children from 1328 schools across the United States, selected through a multistage probability sampling process and spanning kindergarten through 5th grade. Using survival analysis with the dependent variable as the time required to attain a specific milestone, the study revealed that boys, non-Black, non-Hispanic, and high-income children reached the science learning threshold earlier than their counterparts—girls, Black, Hispanic, and low-income children. Furthermore, the study underscored the crucial role of language and literacy skills in mitigating these disparities. The study’s implications stress the importance of targeted interventions to address early science education disparities, emphasizing professional development and integrating language and literacy with science learning. The research also enriches the global discourse on educational standards by introducing innovative methodologies to assess both the frequency and duration of science learning milestones.

Abstract In ovarian cancer, there is no current method to accurately predict recurrence after a complete response to chemotherapy. Here, we develop a machine learning risk score using serum proteomics for the prediction of early recurrence of ovarian cancer after initial treatment. The developed risk score was validated in an independent cohort with serum collected prospectively during the remission period. In the discovery cohort, patients scored as low-risk had a median time to recurrence (TTR) that was not reached at 10 years compared to 10.5 months (HR 4.66, p

Sustained compressive injury (SCI) in the brain is observed in numerous injury and pathological scenarios, including tumors, ischemic stroke, and traumatic brain injury-related tissue swelling. Sustained compressive injury is characterized by tissue loading over time, and currently, there are few in vitro models suitable to study neural cell responses to strain-dependent sustained compressive injury. Here, we present an in vitro model of sustained compressive neural injury via centrifugation. Spheroids were made from neonatal rat cortical cells seeded at 4000 cells/spheroid and cultured for 14 days in vitro. A subset of spheroids was centrifuged at 104, 209, 313 or 419 rads/s for 2 minutes. Modeling the physical deformation of the spheroids via finite element analyses, we found that spheroids centrifuged at the aforementioned angular velocities experienced pressures of 10, 38, 84 and 149 kPa, respectively, and compressive (resp. tensile) strains of 10% (5%), 18% (9%), 27% (14%) and 35% (18%), respectively. Quantification of LIVE-DEAD assay and Hoechst 33342 nuclear staining showed that centrifuged spheroids subjected to pressures above 10 kPa exhibited significantly higher DNA damage than control spheroids at 2, 8, and 24 hours post-injury. Immunohistochemistry of β3-tubulin networks at 2, 8, and 24 hours post-centrifugation injury showed increasing degradation of microtubules over time with increasing strain. Our findings show that cellular injuries occur as a result of specific levels and timings of sustained tissue strains. This experimental SCI model provides a high throughput in vitro platform to examine cellular injury, to gain insights into brain injury that could be targeted with therapeutic strategies.

IntroductionResearch has documented that home and neighborhood contexts of children from low-income families are associated with lower cognitive and social–emotional skills than their higher-income peers. Even though over a third of young children growing up in poverty are dual language learners (DLLs), little research has examined how contextual effects differ between DLL and monolingual children. The current study examines how these two contexts, neighborhood vulnerability and family socioeconomic risk, impact executive function (EF) and social–emotional skills in DLL and monolingual preschoolers.MethodsA secondary analysis was completed on data from two Head Start programs. A series of cross-classified models with interactions were conducted to examine the moderating role of DLL status on associations between neighborhood vulnerability and family risk and preschoolers’ EF and social-emotional skills.ResultsProficient bilingual children’s EF skills were not impacted by neighborhood risks, suggesting that proficient bilingual children may have more opportunities to grow their EF skills when switching between English and Spanish regardless of neighborhood context. An unexpected result occurred for emergent bilingual children who were reported to demonstrate fewer behavior problems regardless of family risk, highlighting the importance of ensuring all DLL families have access to resources to promote their children’s social–emotional skills; and teachers have the proper training to support the behaviors of children in their classroom with varying levels of English proficiency.DiscussionAlthough speaking two languages may be a protective factor for young DLLs growing up in poverty, little research has examined how contextual effects differ between DLL and monolingual children. The current study contributes by examining how DLL status, especially two different DLL statuses (i.e., Proficient Bilinguals and Emergent Bilinguals), may vary as a buffer in moderating the negative associations between collective neighborhood vulnerability, individual family risk, and children’s EF and social–emotional skills.

Type I diabetes is characterized by autoantibodies directed against protein or non-protein self-antigens. Here the authors profile glycan reactive anti-carbohydrate antibodies (ACA) in a longitudinal and cross-sectional childhood diabetes cohort and associate clusters of ACA with disease progression.

Interleukin-1 antagonism is a proposed biomarker and potential therapy for the delay and/or treatment of type 1 diabetes (T1D). We evaluated the role of circulating interleukin-1 receptor antagonist (IL-1ra) in a prospectively monitored cohort of T1D patients. In order to determine a mechanistic association between IL-1ra and T1D, we performed co-localization analyses between serum IL-1ra protein quantitative trait loci and T1D genome-wide analysis studies. Adjusting for human leukocyte antigen (HLA) genotypes, first degree relative status, gender, and age, serum levels of IL-1ra were lower in subjects who progressed to T1D compared to the controls (p = 0.023). Our results suggest that females have higher levels of IL-1ra compared to males (p = 0.005). The 2q14.1 region associated with serum IL-1ra levels is not associated with a risk of developing T1D. Our data suggest that IL-1 antagonism by IL-1ra is not an effective therapy in T1D, but IL-1ra may be a biomarker for progression to T1D.

Nasopharyngeal colonization by Streptococcus pneumoniae is a prerequisite for pneumococcal transmission and disease. Current vaccines protect only against disease and colonization caused by a limited number of serotypes, consequently allowing serotype replacement and transmission. Therefore, the development of a broadly protective vaccine against colonization, transmission and disease is desired but requires a better understanding of pneumococcal adaptation to its natural niche. Hence, we measured the levels of free and protein-bound transition metals in human nasal fluid, to determine the effect of metal concentrations on the growth and proteome of S. pneumoniae. Pneumococci cultured in medium containing metal levels comparable to nasal fluid showed a highly distinct proteomic profile compared to standard culture conditions, including the increased abundance of nine conserved, putative surface-exposed proteins. AliA, an oligopeptide binding protein, was identified as the strongest protective antigen, demonstrated by the significantly reduced bacterial load in a murine colonization and a lethal mouse pneumonia model, highlighting its potential as vaccine antigen.

Exposure to teachers and teaching-related activities is vital for young children’s learning. When COVID-19 closed schools, teachers responded with a mix of live- and prerecorded lessons and one-on-one communication with students, which necessitated shifts in planning time. The current study identifies pre-COVID predictors of time teachers devoted to each of these teaching-related activities to illuminate actionable levers for supporting educators during widespread educational disruption. Teachers with higher prepandemic job commitment devoted more overall time to pandemic-induced remote teaching. Teachers’ prepandemic executive functioning and observed classroom instructional and organizational quality—features of successful teachers during normal times—predicted more during-pandemic time remote teaching, while teacher older age and having more high-needs students was associated with less remote teaching time. These results contribute to an emerging literature that spotlights potential promising avenues for supporting teachers via professional development during normal times as well as in future widespread educational disruptions.

Objectives In lupus nephritis the pathological diagnosis from tissue retrieved during kidney biopsy drives treatment and management. Despite recent approval of new drugs, complete remission rates remain well under aspirational levels, necessitating identification of new therapeutic targets by greater dissection of the pathways to tissue inflammation and injury. This study assessed the safety of kidney biopsies in patients with SLE enrolled in the Accelerating Medicines Partnership, a consortium formed to molecularly deconstruct nephritis.Methods 475 patients with SLE across 15 clinical sites in the USA consented to obtain tissue for research purposes during a clinically indicated kidney biopsy. Adverse events (AEs) were documented for 30 days following the procedure and were determined to be related or unrelated by all site investigators. Serious AEs were defined according to the National Institutes of Health reporting guidelines.Results 34 patients (7.2%) experienced a procedure-related AE: 30 with haematoma, 2 with jets, 1 with pain and 1 with an arteriovenous fistula. Eighteen (3.8%) experienced a serious AE requiring hospitalisation; four patients (0.8%) required a blood transfusion related to the kidney biopsy. At one site where the number of cores retrieved during the biopsy was recorded, the mean was 3.4 for those who experienced a related AE (n=9) and 3.07 for those who did not experience any AE (n=140). All related AEs resolved.Conclusions Procurement of research tissue should be considered feasible, accompanied by a complication risk likely no greater than that incurred for standard clinical purposes. In the quest for targeted treatments personalised based on molecular findings, enhanced diagnostics beyond histology will likely be required.

For decades, Mycobacterium tuberculosis

Objectives Social engagement benefits emotional well‐being, but the mechanisms need to be better understood. Using volunteering as an example of social engagement, we first examined whether volunteering would be longitudinally beneficial to life satisfaction in older adulthood and the moderating role of changes in the social network size in such a relationship as a potential mechanism. Second, we investigated motivations to volunteer in order to promote more volunteering in older adulthood. Methods A sample of 1,591 older participants from the Personality and Total Health (PATH) through life study (PATH) was assessed (Age: Mean = 66.54-years, SD = 1.50-years, 48% female). PATH is a four‐wave longitudinal project with a 4‐year interval between each two waves. Data from PATH Waves 3 and 4 were used to examine the relationship between volunteering and life satisfaction changes and the moderating role of changes in the social network size. Data from PATH Waves 2, 3, and 4 were used to examine the motivations to short‐term and long‐term volunteer. Results and Conclusion We found that longer volunteering time was associated with a higher level of increase in life satisfaction during the 4‐year interval. Life satisfaction increased more in participants who lost more friends than in participants who lost fewer friends. Consistent with socioemotional selectivity theory, emotional goals were positively associated with more short‐term and long‐term volunteering behaviours. These findings provide scientific insight into the mechanisms and motives underlying the positive effects of volunteering on life satisfaction in old age.

Chronic low-grade inflammation is involved in the pathogenesis of type-1 diabetes (T1D) and its complications. In this cross-section study design, we investigated association between serum levels of soluble cytokine receptors with presence of peripheral neuropathy in 694 type-1 diabetes patients. Sex, age, blood pressure, smoking, alcohol intake, HbA1c and lipid profile, presence of DPN (peripheral and autonomic), retinopathy and nephropathy was obtained from patient’s chart. Measurement of soluble cytokine receptors, markers of systemic and vascular inflammation was done using multiplex immunoassays. Serum levels were elevated in in DPN patients, independent of gender, age and duration of diabetes. Crude odds ratios were significantly associated with presence of DPN for 15/22 proteins. The Odds ratio (OR) remained unchanged for sTNFRI (1.72, p=0.00001), sTNFRII (1.45, p=0.0027), sIL2Rα (1.40, p=0.0023), IGFBP6 (1.51, p=0.0032) and CRP (1.47, p=0.0046) after adjusting for confounding variables, HbA1C, hypertension and dyslipidemia. Further we showed risk of DPN is associated with increase in serum levels of sTNFRI (OR=11.2, p

Multiple cross-sectional and longitudinal studies have shown that serum levels of the chemokine ligand 2 (CCL-2) are associated with type 1 diabetes (T1D), although the direction of effect differs. We assessed CCL-2 serum levels in a longitudinal cohort to clarify this association, combined with genetic data to elucidate the regulatory role of CCL-2 in T1D pathogenesis.The Diabetes Autoimmunity Study in the Young (DAISY) followed 310 subjects with high risk of developing T1D. Of these, 42 became persistently seropositive for islet autoantibodies but did not develop T1D (non-progressors); 48 did develop T1D (progressors). CCL-2 serum levels among the three study groups were compared using linear mixed models adjusting for age, sex, HLA genotype, and family history of T1D. Summary statistics were obtained from the CCL-2 protein quantitative trait loci (pQTL) and CCR2 expression QTL (eQTL) studies. The T1D fine mapping association data were provided by the Type 1 Diabetes Genetics Consortium (T1DGC).Serum CCL-2 levels were significantly lower in both progressors (p = 0.004) and non-progressors (p = 0.005), compared to controls. Two SNPs (rs1799988 and rs746492) in the 3p21.31 genetic locus, which includes the CCL-2 receptor, CCR2, were associated with increased CCR2 expression (p = 8.2e-5 and 5.2e-5, respectively), decreased CCL-2 serum level (p = 2.41e-9 and 6.21e-9, respectively), and increased risk of T1D (p = 7.9e-5 and 7.9e-5, respectively).The 3p21.31 genetic region is associated with developing T1D through regulatory control of the CCR2/CCL2 immune pathway.

BackgroundIn this phase I/II trial, 5-year physician-assessed toxicity and patient reported quality of life data is reported for patients undergoing moderately hypofractionated intensity modulated radiation therapy (IMRT) for prostate cancer using a simultaneous integrated boost (SIB) and pelvic lymph node (LN) coverage.Materials and MethodsPatients with T1-T2 localized prostate cancer were prospectively enrolled, receiving risk group based coverage of prostate ± seminal vesicles (SVs) ± pelvic lymph nodes (LNs). Low risk (LR) received 69.6 Gy/29 fractions to the prostate, while intermediate risk (IR) and high risk (HR) patients received 72 Gy/30fx to the prostate and 54Gy/30fx to the SVs. If predicted risk of LN involvement >15%, 50.4 Gy/30fx was delivered to pelvic LNs. Androgen deprivation therapy was given to IR and HR patients.ResultsThere were 55 patients enrolled and 49 patients evaluable at a median follow up of 60 months. Included were 11 (20%) LR, 23 (41.8%) IR, and 21 (38.2%) HR patients. Pelvic LN treatment was given in 25 patients (51%). Prevalence rates of late grade 2 GI toxicity at 1, 3, and 5 years was 5.8, 3.9, and 5.8%, respectively, with no permanent grade 3 events. Prevalence rates of late grade 2 GU toxicity at 1, 3, and 5 years rates were 15.4, 7.7, and 13.5%, respectively, with three grade 3 events (5.8%). The biochemical relapse free survival at 5 years was 88.3%. There were no local, regional, or distant failures, with all patients still alive at last follow up.ConclusionModerate hypofractionation of localized prostate cancer utilizing a SIB technique and LN coverage produces tolerable acute/late toxicity. Given equivalent efficacy between moderate hypofractionation schedules, the optimal regimen will be determined by long-term toxicity reported from both the physician and patient perspective.Clinical Trial Registrationwww.ClinicalTrials.gov, identifier NCT01117935, Date of Registration: 5/6/2010.

Abstract CSF‐1 is a key factor in regulating bone remodeling; osteocytes express CSF‐1 and its receptor. Viable osteocytes are essential for bone remodeling through cell–cell contact and secretion of factors that regulate osteoblasts and osteoclasts. Increased oxidative stress contributes to osteocyte death and correlates with bone loss during aging. The NADPH oxidase Nox4 is a major source of ROS in bone. CSF‐1 decreases Nox4, suggesting that CSF‐1 protects against oxidative stress. Here, we show that osteocyte apoptosis previously reported in our global CSF‐1KO mice is associated with increased Nox4, as well as 4‐HNE expression in osteocytes. Osteocytes isolated from CSF‐1KO mice were less viable and showed increased intracellular ROS, elevated NADPH oxidase activity/Nox4 protein, activation of mTOR/S6K, and downstream apoptosis signals compared with WT osteocytes. Nox4 expression was also increased in CSF‐1KO osteocytes and colocalized with MitoTracker Red in mitochondria. Notably, CSF‐1 inhibited Nox4 expression and apoptosis cascade signals. In additional studies, shNox4 decreased these signals in CSF‐1KO osteocytes, whereas overexpression of Nox4 in WT osteocytes activated the apoptosis pathway. To determine the role of CSF‐1 in osteocytes, DMP1Cre‐CSF‐1cKO (CSF‐1cKO) mice that lack CSF‐1 in osteocytes/late osteoblasts were developed. Osteocyte defects in CSF‐1cKO mice overlapped with those in CSF‐1KO mice, including increased apoptosis, Nox4, and 4‐HNE‐expressing osteocytes. CSF‐1cKO mice showed unbalanced cancellous bone remodeling with decreased bone formation and resorption. Continued exposure to high Nox4/ROS levels may further compromise bone formation and predispose to bone loss and skeletal fragility. Taken together, our findings suggest a novel link between CSF‐1, Nox4‐derived ROS, and osteocyte survival/function that is crucial for osteocyte‐mediated bone remodeling. Results reveal new mechanisms by which CSF‐1/oxidative stress regulate osteocyte homeostasis, which may lead to therapeutic strategies to improve skeletal health in aging. © 2018 American Society for Bone and Mineral Research

Bacterial outer membrane vesicles (OMVs) attract increasing interest as immunostimulatory nanoparticles for the development of vaccines and therapeutic agents. We previously engineered the autotransporter protein Hemoglobin protease (Hbp) into a surface display carrier that can be expressed to high density on the surface of Salmonella OMVs. Moreover, we implemented Tag-Catcher protein ligation technology, to obtain dense display of single heterologous antigens and nanobodies on the OMVs through coupling to the distal end of the Hbp passenger domain. Here, we aimed to further expand the versatility of the Hbp platform by enabling the coupling of heterologous proteins to internal sites of the Hbp passenger. Inserted SpyTags were shown to be accessible at the Salmonella OMV surface and to efficiently couple SpyCatcher-equipped fusion proteins. Next, we combined distally placed SnoopCatcher or SnoopTag sequences with internal SpyTags in a single Hbp molecule. This allowed the coupling of two heterologous proteins to a single Hbp carrier molecule without obvious steric hindrance effects. Since coupling occurs to Hbp that is already exposed on the OMVs, there are no limitations to the size and complexity of the partner proteins. In conclusion, we constructed a versatile modular platform for the development of bivalent recombinant OMV-based vaccines and therapeutics.

Abstract Background Successful nerve regeneration depends upon directed migration of morphologically specialized repair state Schwann cells across a nerve defect. Although several groups have studied directed migration of Schwann cells in response to chemical or topographic cues, the current understanding of how the mechanical environment influences migration remains largely understudied and incomplete. Therefore, the focus of this study was to evaluate Schwann cell migration and morphodynamics in the presence of stiffness gradients, which revealed that Schwann cells can follow extracellular gradients of increasing stiffness, in a form of directed migration termed durotaxis. Methods Polyacrylamide substrates were fabricated to mimic the range of stiffness found in peripheral nerve tissue. We assessed Schwann cell response to substrates that were either mechanically uniform or embedded with a shallow or steep stiffness gradient, respectively corresponding to the mechanical niche present during either the fluid phase or subsequent matrix phase of the peripheral nerve regeneration process. We examined cell migration (velocity and directionality) and morphology (elongation, spread area, nuclear aspect ratio, and cell process dynamics). We also characterized the surface morphology of Schwann cells by scanning electron microscopy. Results On laminin-coated polyacrylamide substrates embedded with either a shallow (∼0.04 kPa/mm) or steep (∼0.95 kPa/mm) stiffness gradient, Schwann cells displayed durotaxis, increasing both their speed and directionality along the gradient materials, fabricated with elastic moduli in the range found in peripheral nerve tissue. Uniquely and unlike cell behavior reported in other cell types, the durotactic response of Schwann cells was not dependent upon the slope of the gradient. When we examined whether durotaxis behavior was accompanied by a pro-regenerative Schwann cell phenotype, we observed altered cell morphology, including increases in spread area and the number, elongation, and branching of the cellular processes, on the steep but not the shallow gradient materials. This phenotype emerged within hours of the cells adhering to the materials and was sustained throughout the 24 hour duration of the experiment. Control experiments also showed that unlike most adherent cells, Schwann cells did not alter their morphology in response to uniform substrates of different stiffnesses. Conclusion This study is notable in its report of durotaxis of cells in response to a stiffness gradient slope, which is greater than an order of magnitude less than reported elsewhere in the literature, suggesting Schwann cells are highly sensitive detectors of mechanical heterogeneity. Altogether, this work identifies durotaxis as a new migratory modality in Schwann cells, and further shows that the presence of a steep stiffness gradient can support a pro-regenerative cell morphology.

Malawian mothers regardless of gestation at birth find pregnancy to be a stressful period that triggers worry due to a fear of maternal death. Research has mostly focused on maternal depressive symptoms regardless of gestation. Limited literature exists on posttraumatic stress symptoms and maternal worry. The purpose of this study was to translate and validate the Perinatal PTSD Questionnaire and Child Health Worry Scale for assessment of posttraumatic stress symptoms and worry about child’s health, respectively, among Malawian women. The process of assuring the accuracy of the translation involved having five native English-speaking nurse researchers and eight Malawian nurse-midwives in a focus group review the translations. We then psychometrically tested the instruments with 30 postpartum mothers at Kamuzu College of Nursing and Queen Elizabeth Central Hospital. Findings from the nurse researchers and the nurse-midwives agreed with most of the translations and modifications were done to improve the adaptations. The instruments were administered to the postpartum mothers. The Chichewa Self Reporting Questionnaire was used as the standard for assessment of perinatal emotional distress symptoms. Both the Chichewa Perinatal PTSD Questionnaire and Chichewa Child Health Worry Scale had high internal consistency and showed strong to moderate correlations with the Chichewa Self Reporting Questionnaire. The mothers agreed that the questions were easy to understand and culturally sensitive, although a few questions were seen as distressing. Future adaptations need to consider testing the instruments in mothers throughout infancy and utilizing other gold standard instruments for validation. Keywords: Depression, Posttraumatic stress, Anxiety, Worry, Mothers, Postpartum period

Purpose: To determine the combined effect of TheraTears® Lubricant Eye Drops, TheraTears® SteriLid Eyelid Cleanser, and TheraTears® Nutrition on dry eye signs and symptoms. Methods: This prospective study enrolled 28 dry eye participants. Participants were instructed to use the Lubricant Eye Drops at least 2–4× a day, SteriLid 1–2× a day, and Nutrition 3 gel caps once a day. Participants were followed up at baseline, 1 month and 3 months. Outcome variables were the Ocular Surface Disease Index (OSDI), Symptom Assessment iN Dry Eye (SANDE) questionnaire, non-invasive tear break-up time (NIBUT), osmolarity, number of meibomian glands blocked (#MG blocked), meibum quality, eyelid margin features, Schirmer's test, tear film lipid layer thickness (LLT), meniscus height, corneal and conjunctival staining. Results: Twenty participants (mean age = 43, from 23 to 66, 17F, 3M) completed the study. Participants reported having used, on average, the Lubricant Eye Drop 2.4×/day, the SteriLid 1.1×/day, and the Nutrition 3 gel caps 1×/day. There was a significant change over time (p

The purpose of this study was to determine whether maternal mental and physical health is associated with maternal testosterone and cortisol levels, parenting of very low birth weight infants, physical exercise, and White vs non-White race. A total of 40 mothers of very low birth weight infants were recruited from a neonatal intensive care unit at a University Hospital in the Southeast United States. Data were collected through a review of medical records, standardized questionnaires, and biochemical measurement. Maternal mental and physical health status using questionnaires as well as maternal testosterone and cortisol levels using an enzyme immunoassay were measured four times (birth, 40 weeks postmenstrual age [PMA], and 6 and 12 months [age of infant, corrected age]). General linear models showed that higher testosterone levels were associated with greater depressive symptoms, stress, and poorer physical health at 40 weeks PMA, and at 6 and 12 months. High cortisol levels were associated with greater anxiety at 40 weeks PMA; however, with better mental and physical health at 40 weeks PMA, and 6 and 12 months. Physical activity was associated with lower maternal perceived stress at 12 months. Maternal health did not differ by race, except anxiety, which was higher in White than non-White mothers after birth. As very low birth weight infants grew up, maternal physical health improved but mental health deteriorated. Testosterone and cortisol levels were found to be positively correlated in women but testosterone was more predictive of maternal mental and physical health than cortisol. Indeed testosterone consistently showed its associations with maternal health. Maternal stress might be improved through regular physical exercise.

Continuous positive airway pressure (CPAP) therapy is very often the treatment of choice for obstructive sleep apnoea (OSA). The association between intracranial hypertension and spontaneous cerebrospinal fluid (CSF) rhinorrhoea is being increasingly recognized among patients with OSA. However, spontaneous CSF rhinorrhoea following initiation of CPAP therapy for OSA is very rarely documented in the literature. In this report, we describe a 53‐year‐old woman with severe OSA who, while being evaluated for possible intracranial hypertension, developed spontaneous CSF rhinorrhoea and CSF aspiration pneumonitis as a complication of CPAP therapy. Magnetic resonance imaging confirmed fluid tracks at the skull base, and a nasal swab demonstrated positive β2‐transferrin. Computer tomography (CT) chest showed findings consistent with CSF aspiration pneumonitis. Resolution of both CSF leak and pneumonitis were noted following treatment with azetozolamide and curative endoscopic trans‐nasal surgery along with ventriculoperitoneal shunt.

Outer membrane vesicles (OMVs) are promising vaccine components because they combine antigen and adjuvant in a single formulation. Detoxified Salmonella enterica strains that express penta-acylated lipid A retain OMV immunogenicity but with reduced reactogenicity. We have previously shown that a recombinant form of the enterotoxigenic Escherichia coli (ETEC) 17 kilodalton protein (Skp) protects mice in a pulmonary challenge model, when fused to the glutathione-S-transferase (GST) epitope and combined with cholera toxin. Here we compared directly the efficacy of expressing Skp in detoxified Salmonella OMVs to GST-Skp for their ability to protect mice against ETEC challenge. We observed that the display of Skp on OMVs, in the absence of exogenous adjuvant, protects the mice as well as the recombinant GST-Skp with adjuvant, showing that we can achieve protection when antigen and adjuvant are administered as a single formulation. Collectively, these data demonstrate the utility of using OMVs for the expression and display of antigens for use in vaccine development and validate previously published work demonstrating that immunization with Skp is efficacious in protecting mice against ETEC challenge.

Soluble cytokine receptors may play an important role in development of microalbuminuria (MA) in type-1 diabetes (T1D). In this study, we measured 12 soluble receptors and ligands from TNF-α/IL6/IL2 pathways in T1D patients with MA (n = 89) and T1D patients without MA (n = 483) participating in the PAGODA study. Twelve proteins in the sera from T1D patients with and without MA were measured using multiplex Luminex assays. Ten serum proteins (sTNFR1, sTNFR2, sIL2Rα, MMP2, sgp130, sVCAM1, sIL6R, SAA, CRP, and sICAM1) were significantly elevated in T1D patients with MA. After adjusting for age, duration of diabetes, and sex in logistic regression, association remained significant for seven proteins. MA is associated with increasing concentrations of all 10 proteins, with the strongest associations observed for sTNFR1 (OR = 108.3, P

PTS symptoms are a common negative emotional response of mothers of preterm infants. PTS symptoms are one of the least explored emotional responses in postpartum period and in mothers of preterm infants. Malawi has the highest preterm birth in the world, but little is known about PTS symptoms following preterm birth in Malawi. The purpose of this paper is to review evidence on the incidence, prevalence, and impact of PTS symptoms following preterm birth, predictors of PTS symptoms, screening and management, and to identify the gaps and the applicability of the evidence to developing countries such as Malawi. A literature search was conducted using PubMed, PsychINFO, CINAHL, and ERIC databases. Articles were limited to PTS symptoms in mothers after preterm birth and up to 24 months. A total of 23 articles were included in the analysis. Findings showed that most literature was from developed countries. Fifteen instruments were identified and the PPQ was the most commonly used instrument. Time points for measurement varied. Mothers of preterm infants presented with at least one PTS symptom and they had higher PTS symptoms than mothers of healthy full-term infants, but no significant differences were seen with mothers of sick full term infants. Maternal, infant, and external factors predicted the onsets of PTS symptoms. Psychoeducation and counseling significantly reduced PTS symptoms, although mothers were only referred if symptoms were severe. Research in developing or low-income countries like Malawi is needed and researchers need to engage more in longitudinal approaches.

Vascular risk factors for age-related cognitive decline are significant, and their management may ultimately prove the most successful strategy for reducing risk and sustaining cognitive health. This randomized, double-blinded, placebo-controlled trial with parallel group allocation to either marine n-3 polyunsaturated fatty acids (n-3 PUFA) or soybean oil placebo assesses the effects on the total volume of accumulation in cerebral white matter hyperintensities (WMH), a potentially modifiable neurovascular component of age-related cognitive decline. Total WMH accumulation over 3 years is the primary endpoint. The safety and efficacy of n-3 PUFA is evaluated in older adults with significant WMH and suboptimum plasma n-3 PUFA as inclusion criteria. One hundred and two non-demented older adults were enrolled with a mean age of 81.1 (±4.4) years, WMH of 19.4 (±16.1) cm3, and a plasma n-3 PUFA of 86.64 (±29.21) µg/mL. 61% were female, 28% were apolipoprotein E epsilon 4 carriers, and the mean mini-mental state exam (MMSE) was 27.9 (±1.7). This trial provides an initial evaluation of n-3 PUFA effects on WMH, a reproducible and valid risk biomarker for cognitive decline, as well as on inflammatory biomarkers thought to play a role in WMH accumulation. We present the baseline results and operational experience of enriching a study population on advanced age, blood n-3 PUFA, and magnetic resonance imaging (MRI) derived WMH with biomarker outcomes (WMH, inflammation markers) in a dementia prevention paradigm.

Aims. To examine the association of the serum levels of TNF receptors, adhesion molecules, and inflammatory mediators with diabetic retinopathy (DR) in T1D patients. Methods. Using the multiplex immunoassay, we measured serum levels of eight proteins in 678 T1D subjects aged 20–75 years. Comparisons were made between 482 T1D patients with no complications and 196 T1D patients with DR. Results. The levels of sTNFR-I, sTNFR-II, CRP, SAA, sgp130, sIL6R, sVCAM1, and sICAM1 were significantly higher in the T1D patients with DR as compared to T1D patients with no complications. Multivariate logistic regression analysis revealed significant association for five proteins after adjustment for age, sex, and disease duration (sTNFR-I: OR=1.57, sgp130: OR=1.43, sVCAM1: OR=1.27, sICAM1: OR=1.42, and CRP: OR=1.15). Conditional logistic regression on matched paired data revealed that subjects in the top quartile for sTNFR-I (OR=2.13), sTNFR-II (OR=1.66), sgp130 (OR=1.82), sIL6R (OR=1.75), sVCAM1 (OR=1.98), sICAM1 (OR=2.23), CRP (OR=2.40) and SAA (OR=2.03), had the highest odds of having DR. Conclusions. The circulating markers of inflammation, endothelial injury, and TNF signaling are significantly associated with DR in patients with T1D. TNFR-I and TNFR-II receptors are highly correlated, but DR associated more strongly with TNFR-I in these patients.

Traction Force Microscopy (TFM) is a powerful approach for quantifying cell-material interactions that over the last two decades has contributed significantly to our understanding of cellular mechanosensing and mechanotransduction. In addition, recent advances in three-dimensional (3D) imaging and traction force analysis (3D TFM) have highlighted the significance of the third dimension in influencing various cellular processes. Yet irrespective of dimensionality, almost all TFM approaches have relied on a linear elastic theory framework to calculate cell surface tractions. Here we present a new high resolution 3D TFM algorithm which utilizes a large deformation formulation to quantify cellular displacement fields with unprecedented resolution. The results feature some of the first experimental evidence that cells are indeed capable of exerting large material deformations, which require the formulation of a new theoretical TFM framework to accurately calculate the traction forces. Based on our previous 3D TFM technique, we reformulate our approach to accurately account for large material deformation and quantitatively contrast and compare both linear and large deformation frameworks as a function of the applied cell deformation. Particular attention is paid in estimating the accuracy penalty associated with utilizing a traditional linear elastic approach in the presence of large deformation gradients.

Osteoporosis is a silent disease, characterized by a porous bone micro-structure that enhances risk for fractures and associated disabilities. Senile, or age-related osteoporosis (SO), affects both men and women, resulting in increased morbidity and mortality. However, cellular and molecular mechanisms underlying senile osteoporosis are not fully known. Recent studies implicate the accumulation of reactive oxygen species (ROS) and increased oxidative stress as key factors in SO. Herein, we show that loss of caspase-2, a cysteine aspartate protease involved in oxidative stress-induced apoptosis, results in total body and femoral bone loss in aged mice (20% decrease in bone mineral density), and an increase in bone fragility (30% decrease in fracture strength). Importantly, we demonstrate that genetic ablation or selective inhibition of caspase-2 using zVDVAD-fmk results in increased numbers of bone-resorbing osteoclasts and enhanced tartrate-resistant acid phosphatase (TRAP) activity. Conversely, transfection of osteoclast precursors with wild type caspase-2 but not an enzymatic mutant, results in a decrease in TRAP activity. We demonstrate that caspase-2 expression is induced in osteoclasts treated with oxidants such as hydrogen peroxide and that loss of caspase-2 enhances resistance to oxidants, as measured by TRAP activity, and decreases oxidative stress-induced apoptosis of osteoclasts. Moreover, oxidative stress, quantified by assessment of the lipid peroxidation marker, 4-HNE, is increased in Casp2-/- bone, perhaps due to a decrease in antioxidant enzymes such as SOD2. Taken together, our data point to a critical and novel role for caspase-2 in maintaining bone homeostasis by modulating ROS levels and osteoclast apoptosis during conditions of enhanced oxidative stress that occur during aging.

Nitrogen remains a ubiquitous pollutant in surface and groundwater throughout the United States, despite 30 years of pollution control efforts. A detailed multicompartment N balance for the Central Arizona-Phoenix ecosystem is used to illustrate how an ecosystem-level approach can be used to develop improved N management strategies. The N balance is used to demonstrate how nitrate in pumped groundwater used for crop irrigation could be used to reduce inputs of commercial fertilizer and decrease N leaching to aquifers. Effectively managing N pollution also will require an understanding of the complex factors that control the N balance, including targeted regulations, individual human behavior, land-use conversion, and other ecosystem management practices that affect the N balance. These sometimes countervailing factors are illustrated with several scenarios of wastewater treatment technology and population growth in the Phoenix area. Management of N eventually must be coupled to management of other elements, notably carbon, phosphorus, and salts. We postulate that an ecosystem framework for pollution management will result in strategies that are more effective, fairer, and less expensive than current approaches.

BACKGROUND: Biomarkers play critical roles in early detection, diagnosis and monitoring of therapeutic outcome and recurrence of cancer. Previous biomarker research on ovarian cancer (OC) has mostly focused on the discovery and validation of diagnostic biomarkers. The primary purpose of this study is to identify serum biomarkers for prognosis and therapeutic outcomes of ovarian cancer. EXPERIMENTAL DESIGN: Forty serum proteins were analyzed in 70 serum samples from healthy controls (HC) and 101 serum samples from serous OC patients at three different disease phases: post diagnosis (PD), remission (RM) and recurrence (RC). The utility of serum proteins as OC biomarkers was evaluated using a variety of statistical methods including survival analysis. RESULTS: Ten serum proteins (PDGF-AB/BB, PDGF-AA, CRP, sFas, CA125, SAA, sTNFRII, sIL-6R, IGFBP6 and MDC) have individually good area-under-the-curve (AUC) values (AUC = 0.69-0.86) and more than 10 three-marker combinations have excellent AUC values (0.91-0.93) in distinguishing active cancer samples (PD & RC) from HC. The mean serum protein levels for RM samples are usually intermediate between HC and OC patients with active cancer (PD & RC). Most importantly, five proteins (sICAM1, RANTES, sgp130, sTNFR-II and sVCAM1) measured at remission can classify, individually and in combination, serous OC patients into two subsets with significantly different overall survival (best HR = 17, p

Chemokine (C-C motif) ligand 2 (CCL2), commonly known as monocyte chemoattractant protein-1 (MCP-1), has been implicated in the pathogenesis of many diseases characterized by monocytic infiltration. However, limited data have been reported on MCP-1 in type 1 diabetes (T1D) and the findings are inconclusive and inconsistent.In this study, MCP-1 was measured in the sera from 2,472 T1D patients and 2,654 healthy controls using a Luminex assay. The rs1024611 SNP in the promoter region of MCP-1 was genotyped for a subset of subjects (1764 T1D patients and 1323 controls) using the TaqMan-assay.Subject age, sex or genotypes of MCP-1 rs1024611SNP did not have a major impact on serum MCP-1 levels in either healthy controls or patients. While hemoglobin A1c levels did not have a major influence on serum MCP-1 levels, the mean serum MCP-1 levels are significantly higher in patients with multiple complications (mean = 242 ng/ml) compared to patients without any complications (mean = 201 ng/ml) (p = 3.5×10(-6)). Furthermore, mean serum MCP-1 is higher in controls (mean = 261 ng/ml) than T1D patients (mean = 208 ng/ml) (p99(th) percentile of patients or 955 ng/ml) of serum MCP-1 is significantly lower in the T1D group compared to the control group (odds ratio = 0.11, p

Directed migration of Schwann cells (SC) is critical for development and repair of the peripheral nervous system. Understanding aspects of motility specific to SC, along with SC response to engineered biomaterials, may inform strategies to enhance nerve regeneration. Rat SC were cultured on laminin-coated microgrooved poly(dimethyl siloxane) platforms that were flat or presented repeating cellular scale anisotropic topographical cues, 30 or 60 µm in width, and observed with timelapse microscopy. SC motion was directed parallel to the long axis of the topography on both the groove floor and the plateau, with accompanying differences in velocity and directional persistence in comparison to SC motion on flat substrates. In addition, feature dimension affected SC morphology, alignment, and directional persistence. Plateaus and groove floors presented distinct cues which promoted differential motility and variable interaction with the topographical features. SC on the plateau surfaces tended to have persistent interactions with the edge topography, while SC on the groove floors tended to have infrequent contact with the corners and walls. Our observations suggest the capacity of SC to be guided without continuous contact with a topographical cue. SC exhibited a range of distinct motile morphologies, characterized by their symmetry and number of extensions. Across all conditions, SC with a single extension traveled significantly faster than cells with more or no extensions. We conclude that SC motility is complex, where persistent motion requires cellular asymmetry, and that anisotropic topography with cellular scale features can direct SC motility.

Type 1 diabetes (T1D) is an autoimmune disease resulting from the complex interaction between multiple susceptibility genes, environmental factors and the immune system. Over 40 T1D susceptibility regions have been suggested by recent genome-wide association studies; however, the specific genes and their role in the disease remain elusive. The objective of this study is to identify the susceptibility gene(s) in the 12q13 region and investigate the functional link to the disease pathogenesis. A total of 19 SNPs in the 12q13 region were analyzed by the TaqMan assay for 1,434 T1D patients and 1,865 controls. Thirteen of the SNPs are associated with T1D (best p = 4x10(-11)), thus providing confirmatory evidence for at least one susceptibility gene in this region. To identify candidate genes, expression of six genes in the region was analyzed by real-time RT-PCR for PBMCs from 192 T1D patients and 192 controls. SNP genotypes in the 12q13 region are the main factors that determine ERBB3 mRNA levels in PBMCs. The protective genotypes for T1D are associated with higher ERBB3 mRNA level (p

We present evidence that there can be substantial variation in species richness in residential areas differing in their socioeconomic and cultural characteristics. Many analyses of the impacts of urbanization on biodiversity rely on traditional "urban-to-rural" gradient measures, such as distance from urban center or population density, and thus can fail to account for the ways in which human socioeconomic and cultural characteristics are shaping the human-environment interaction and ecological outcomes. This influence of residential values and economic resources on biodiversity within the urban matrix has implications for human quality of life, for urban conservation strategies, and for urban planning.

En el presente trabajo se ofrece una descripción de una batalla ritual, ocurrida en 1772 de la época colonial, entre las mitades de una comunidad de la sierra sur del Perú. Las limitaciones de un proceso criminal, característica esencial de un documento de esta naturaleza, demanda una comparación necesaria de los elementos constitutivos de dicha batalla con otros conflictos similares que sucedieron durante el transcurso del tiempo y el espacio. Este análisis comparativo permite una mayor perspectiva en el enfoque de la investigación, de manera tal que las batallas rituales sean ubicadas de una manera más general en un contexto cultural y simbólico específico. Una serie de elementos simbólicos invariantes se extraen de un extenso ciclo ritual. Estos elementos se concentran en la fertilidad de la tierra, animales domésticos y seres humanos, los cuales se vinculan en una relación de reciprocidad. Asimismo, se enfatiza en factores tales como la separación de grupos sociales, el intercambio y los límites. Finalmente, se considera la palabra quechua pukllay (o "juego") para referirse al ciclo total, y se la vincula provisionalmente al énfasis en el delinamiento de grupos y límites.