To establish a standardized, trauma informed and family-centered emergency department (ED) sudden and unexpected infant death (SUID) management protocol at Nemours Children's Health, Delaware for ...medical professionals and multidisciplinary team (MDT) collaborators, informed by national clinical practice guidance, and respective of both family and investigative needs. SUID are emotionally distressing for involved family members, often precipitated by profound grief and confusion as the family interacts with many mandated public agencies during the course of a medicolegal death investigation. Although SUID necessitates consideration of child abuse and neglect as a contributory factor, and accurate determination of death cause may have critical implications for other family members and public health, prioritizing family needs in a trauma informed manner is paramount. Collaboration between MDT partners to provide optimal care to families following SUID involves transparent family communication, attending to medical and mental health needs of surviving family (especially siblings), and respecting medicolegal investigative constraints. Many institutions lack standardized approaches to SUID cases, which may precipitate increased family distress and delay initiation of necessary medicolegal death procedures.
An MDT expert panel consisting of medical, legal, law enforcement, and child welfare professionals was convened at Nemours Children's Health, Delaware in 2018 over a 3-month period to analyze and implement an enhanced, family-centered, trauma informed hospital protocol.
Using exploratory inquiry and dialogue to elicit important protocol goals, a family-centered protocol with revised, coordinated roles for MDT members was developed with enhanced focus on communication, family-, and team-oriented care.
Implementation of a family-centered, ED-based protocol standardizing the approach to SUID effectively supports medicolegal death investigative procedures while prioritizing trauma informed, supportive, sensitive ED care for grieving families.
Health care institutions serving children and their families should develop and implement trauma informed, family-centered protocols to ensure sensitivity during medicolegal death investigations.
Facilitated glucose transport across plasma membranes is mediated by a family of transporters (GLUT1-GLUT5) that have different tissue distributions and Km values for transport. It has been shown ...that insulin stimulates glucose transport in fat and muscle tissues by causing the redistribution of one of these proteins (GLUT4) from inside the cell to the plasma membrane. Previous studies have shown that agents that change cAMP levels are able to modulate glucose transport in fat cells. The aim of this study was to investigate the mechanisms responsible for modulation of glucose transport by cAMP. 2-Deoxyglucose transport and insulin-regulatable glucose transporter (GLUT4) immunoreactivity in plasma and low density microsomal membranes were measured in adipocytes incubated for 30 min with insulin or dibutyryl-cAMP (Bt2cAMP). Low concentrations of Bt2cAMP (10 microM) increased 2-deoxyglucose uptake by translocating GLUT4 from low density microsomal membranes to the plasma membranes. Bt2cAMP at 1000 microM inhibited glucose transport below basal but further increased translocation of GLUT4. The effect of Bt2cAMP on translocation was additive to that of 7 nM insulin. We conclude that in rat adipocytes, Bt2cAMP acutely translocates GLUT4 but inhibits its activity to transport glucose.
In adipocytes, insulin stimulates the translocation of the glucose transporter, GLUT4, from an intracellular storage compartment to the cell surface. Substantial evidence exists to suggest that in ...the basal state GLUT4 resides in discrete storage vesicles. A direct interaction of GLUT4 storage vesicles with the plasma membrane has been implicated because the v-SNARE, vesicle-associated membrane protein-2 (VAMP2), appears to be a specific component of these vesicles. In the present study we sought to identify the cognate target SNAREs for VAMP2 in mouse 3T3-L1 adipocytes. Membrane fractions were isolated from adipocytes and probed by far Western blotting with the cytosolic portion of VAMP2 fused to glutathione S-transferase. Two plasma membrane-enriched proteins, p25 and p35, were specifically labeled with this probe. By using a combination of immunoblotting, detergent extraction, and anion exchange chromatography, we identified p35 as Syntaxin-4 and p25 as the recently identified murine SNAP-25 homologue, Syndet (mSNAP-23). By using surface plasmon resonance we show that VAMP2, Syntaxin-4, and Syndet form a ternary SDS-resistant SNARE complex. Microinjection of anti-Syndet antibodies into 3T3-L1 adipocytes, or incubation of permeabilized adipocytes with a synthetic peptide comprising the C-terminal 24 amino acids of Syndet, inhibited insulin-stimulated GLUT4 translocation to the cell surface by approximately 40%. GLUT1 trafficking remained unaffected by the presence of the peptide. Our data suggest that Syntaxin-4 and Syndet are important cell-surface target SNAREs within adipocytes that regulate docking and fusion of GLUT-4-containing vesicles with the plasma membrane in response to insulin.
munc18c is a critical protein involved in trafficking events associated with syntaxin 4 and which also mediates inhibitory effects on vesicle docking and/or fusion. To investigate the domains of ...munc18c responsible for its interaction with syntaxin 4, fragments of munc18c were generated and their interaction with syntaxin 4 examined in vivo by the yeast two-hybrid assay. In vitro protein-protein interaction studies were then used to confirm that the interaction between the proteins was direct. Full-length munc18c(1-592), munc18c(1-139) and munc18c(1-225), but not munc18c(226-592), munc18c(1-100), munc18c(43-139) or munc18c(66-139), interacted with the cytoplasmic portion of syntaxin 4, Stx4(2-273), as assessed by yeast two-hybrid assay of growth on nutritionally deficient media and by beta-galactosidase reporter induction. The N-terminal predicted helix-a-helix-b-helix-c region of syntaxin 4, Stx4(29-157), failed to interact with full-length munc18c(1-592), indicating that a larger portion of syntaxin 4 is necessary for the interaction. The yeast two-hybrid results were confirmed by protein-protein interaction studies between Stx4(2-273) and glutathione S-transferase fusion proteins of munc18c. Full-length munc18c(1-592), munc18c(1-139) and munc18c(1-225) interacted with Stx4(2-273) whereas munc18c(1-100) did not, consistent with the yeast two-hybrid data. These data thus identify a region of munc18c between residues 1 and 139 as a minimal domain for its interaction with syntaxin 4.
Despite its utility, several conceptual and methodological concerns are raised regarding Fischer and Turner's (1970) Attitudes Toward Seeking Professional Psychological Help Scale (ATSPPHS). These ...concerns were addressed in an adaptation and extension of the ATSPPHS using 208 adult volunteers. The new Inventory of Attitudes Toward Seeking Mental Health Services (IASMHS) consists of 24 items and 3 internally consistent factors: psychological openness, help‐seeking propensity, and indifference to stigma. We replicated this 3‐factor model with 293 university undergraduates, and established test‐retest reliability with 23 student volunteers. Validity was demonstrated by the ability of the IASMHS to distinguish between those who had and had not used mental health services in the past, and those who would and would not use these services in the future. It also discriminated between participants’ intentions to use professional and nonprofessional help. Finally, expected gender differences in help‐seeking attitudes were demonstrated.
In 2019, 4.4 million referrals of maltreatment were made that affected approximately 7.9 million children. It was estimated that 9.3% of the referrals were related to child sexual abuse (CSA). To ...prevent negative psychosocial and health-related outcomes associated with CSA, CSA survivors often participate in a forensic interview, medical and behavioral health assessments, and behavioral health treatment while navigating other life disruptions or changing family dynamics precipitated by the CSA (e.g., change in custody or household, lack of contact with preparator, etc.). The assessment and treatment of pediatric survivors of CSA by multidisciplinary teams (MDT) can enhance families' engagement and participation with the legal process, medical evaluation, and behavioral health services. This paper explores the Nemours Children's Health, Delaware MDT's approach to assessing and treating CSA, explores benefits and barriers associated with the current model, and discusses public health implications of a MDT approach to addressing CSA.
Inhibition of fatty acid (FA) oxidation has been shown previously to lower blood glucose levels acutely in diabetic rats. However, the longer term effects of FA oxidation inhibition have not been ...determined. This study examines the effect of inhibition of (FA) oxidation for 3 weeks on carbohydrate metabolism in rats rendered diabetic with streptozotocin (STZ). STZ treated rats (50 mg/kg) were randomized into 3 groups: a non-treated diabetic group and 2 groups treated with either 12.5 or 25 mg/kg/day Etomoxir (a specific carnitine palmitoyl transferase inhibitor) for 3 weeks. The 3 groups of rats had ad libitum access to a high fat diet (50% energy fat, 30% carbohydrate and 20% protein) throughout the study. After 3 weeks hepatic glucose production (HGP) was estimated using a constant infusion of (3H)-6-glucose in-vivo after an overnight fast. Inhibition of FA oxidation in diabetic rats resulted in a significant reduction in fasting glucose levels and hepatic glucose production. In addition, experiments with adipocytes isolated from diabetic rats treated with etomoxir demonstrated an increase in sensitivity and responsiveness to insulin of glucose utilization and pyruvate dehydrogenase (PDH) activity. It is important to note that these improvements in carbohydrate metabolism were not accompanied by increases in circulating FFA or triglyceride levels which were unchanged or lower after inhibition of FA oxidation.
Human autoantibodies to proteins of the mitotic apparatus have demonstrated clinical utility and usefulness as molecular probes for identification and characterization of novel autoantigens, as ...exemplified by autoantibodies to centromere proteins. In contrast, there have been very few reports of autoantibodies with reactivity to antigens located along mitotic chromosome arms, but not in interphase nuclei. The purpose of this study was to identify and characterize autoantibodies with reactivity to mitotic chromosomal antigens (MCAs) located exclusively on mitotic chromosome arms, and to determine if patients with these autoantibodies have common clinical features.
Routine immunofluorescence screening of serum samples referred for antinuclear antibody investigation over a 10-year period was used to identify autoantibodies to MCAs. MCAs were identified by exclusive immunofluorescence staining of mitotic chromosome arms with no staining of interphase nuclei. MCA-reactive sera were further characterized for patterns of staining on mitotic chromosome arms and sensitivities to chemical and enzymatic treatments, and for one of these sera, its ability to abrogate progression through mitosis when microinjected into cells.
Of 60,000 sera screened for antinuclear antibodies by immunofluorescence, we identified three IgG autoantibodies reacting exclusively to MCAs. The anti-MCA autoantibodies did not react with condensed chromatin in spermatozoa or in apoptotic HeLa cells. Reactivity of all three sera was abrogated by treatment with protease, but not RNase, indicating that the MCAs are protein in nature and do not contain RNA epitopes. The three anti-MCA antibodies seem to react to three different antigens because they gave different patterns of staining of chromosome arms, reacted with chromosomes in different stages of mitosis, and displayed different sensitivities to treatment with DNase 1, salt, and phosphatases. Phosphatase treatment suggests that MCA1 and MCA2 contain serine/threonine phosphoepitope(s) and MCA3 tyrosine phosphoepitope(s). Loss of MCA2 reactivity to DNase 1 treatment and its retention after salt extraction suggests that it is a chromosomal scaffold protein. Sensitivity of all three MCAs to acid suggests that they are histone-like or histone-associated proteins.
We report the identification of three novel MCA-reactive sera. Patient diagnoses included discoid lupus erythematosus, chronic lymphocytic leukemia, Sjögren's syndrome, and polymyalgia rheumatica. The reactivity of anti-MCA antibodies with phosphoepitopes is likely to explain restriction of immunofluorescence staining to chromosome arms during mitosis. Microinjection of MCA1-reactive antibodies led to metaphase arrest, without any change in morphology of the mitotic spindle or metaphase chromosomes suggesting that MCA1 may have a role in sister chromatid separation.
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