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Aortic valve stenosis (AVS) is a progressive fibrotic disease that is caused by thickening and stiffening of valve leaflets. At the cellular level, quiescent valve interstitial cells (qVICs) activate to myofibroblasts (aVICs) that persist within the valve tissue. Given the persistence of myofibroblasts in AVS, epigenetic mechanisms have been implicated. Here, we studied changes that occur in VICs during myofibroblast activation by using a hydrogel matrix to recapitulate different stiffnesses in the valve leaflet during fibrosis. We first compared the chromatin landscape of qVICs cultured on soft hydrogels and aVICs cultured on stiff hydrogels, representing the native and diseased phenotypes respectively. Using assay for transposase-accessible chromatin sequencing (ATAC-Seq), we found that open chromatin regions in aVICs were enriched for transcription factor binding motifs associated with mechanosensing pathways compared to qVICs. Next, we used RNA-Seq to show that the open chromatin regions in aVICs correlated with pro-fibrotic gene expression, as aVICs expressed higher levels of contractile fiber genes, including myofibroblast markers such as alpha smooth muscle actin (αSMA), compared to qVICs. In contrast, chromatin remodeling genes were downregulated in aVICs compared to qVICs, indicating qVICs may be protected from myofibroblast activation through epigenetic mechanisms. Small molecule inhibition of one of these remodelers, CREB Binding Protein (CREBBP), prevented qVICs from activating to aVICs. Notably, CREBBP is more abundant in valves from healthy patients compared to fibrotic valves. Our findings reveal the role of mechanical regulation in chromatin remodeling during VIC activation and quiescence and highlight one potential therapeutic target for treating AVS., Competing Interests: The authors declare no conflicts of interests., (© 2022 The Authors. Bioengineering & Translational Medicine published by Wiley Periodicals LLC on behalf of American Institute of Chemical Engineers.)

Although there has been a significant increase in the delivery of evidence-supported, trauma-informed care over the past few years, there has been less discussion around the consideration of the broader cultural, political, and societal factors that contextualize client trauma and that also need to be recognized and understood to promote healing and prevent future trauma. In support of sharing some best practices and lessons learned, this article provides a case study of one agency that has used the Sanctuary Model ® , an evidence-supported, trauma-informed organizational change model, to introduce the practice of cultural humility with staff as a facilitator of improved service delivery for clients from culturally marginalized communities. The model supports these endeavors through the adherence to the seven commitments, a set of organizational values for creating a trauma-informed community, allowing for all voices to be heard and considered and providing opportunities to begin the repair of previous experiences of inequity and suppression. Through the board of directors, leadership, and staff, the organization transformed its culture into one that truly supports and embraces diversity, equity, and inclusion in its operation for the benefit of both staff and clients alike.

Because of the growing population of patients with serious illness, the demand for specialty palliative care exceeds the resources available. Nurses must be prepared to provide primary palliative care to fill the gap in the availability of specialized palliative care providers. However, meeting the educational needs of a vast number of practicing nurses poses a significant challenge. Often, institutions are limited in the financial and staffing support that they can contribute for continuing nursing education, especially when the training requires staff to spend substantial time away from work. In order to address this issue, one large medical center conducted a study to examine the educational and clinical practice outcomes of offering an online version of the End-of-Life Nursing Education Consortium Core Curriculum to nearly 100 nurses. The participants were divided into 2 groups. Group 1 received online education only, whereas the other group received the online education plus a 3-hour face-to-face training session. Both groups reported statistically significant improvements in symptom management and communication skills, with no significant difference between the pedagogical approaches. Most importantly, the education resulted in a statistically significant impact on the nurses' clinical practice. The results of this study demonstrate that online education can be used as an effective and efficient strategy to provide primary palliative care education to a large number of nurses.

In this work, iminyl σ-radical formation in several one-electron-oxidized cytosine analogs, including 1-MeC, cidofovir, 2'-deoxycytidine (dCyd), and 2'-deoxycytidine 5'-monophosphate (5'-dCMP), were investigated in homogeneous, aqueous (D2O or H2O) glassy solutions at low temperatures by employing electron spin resonance (ESR) spectroscopy. Upon employing density functional theory (DFT) (DFT/B3LYP/6-31G* method), the calculated hyperfine coupling constant (HFCC) values of iminyl σ-radical agree quite well with the experimentally observed ones, thus confirming its assignment. ESR and DFT studies show that the cytosine iminyl σ-radical is a tautomer of the deprotonated cytosine π-cation radical [cytosine π-aminyl radical, C(N4-H)(•)]. Employing 1-MeC samples at various pHs ranging from ca. 8 to 11, ESR studies show that the tautomeric equilibrium between C(N4-H)(•) and the iminyl σ-radical at low temperature is too slow to be established without added base. ESR and DFT studies agree that, in the iminyl σ-radical, the unpaired spin is localized on the exocyclic nitrogen (N4) in an in-plane pure p-orbital. This gives rise to an anisotropic nitrogen hyperfine coupling (Azz = 40 G) from N4 and a near isotropic β-nitrogen coupling of 9.7 G from the cytosine ring nitrogen at N3. Iminyl σ-radical should exist in its N3-protonated form, as the N3-protonated iminyl σ-radical is stabilized in solution by over 30 kcal/mol (ΔG = -32 kcal/mol) over its conjugate base, the N3-deprotonated form. This is the first observation of an isotropic β-hyperfine ring nitrogen coupling in an N-centered DNA radical. Our theoretical calculations predict that the cytosine iminyl σ-radical can be formed in double-stranded DNA by a radiation-induced ionization-deprotonation process that is only 10 kcal/mol above the lowest energy path.

Background and Objective: Consensus nutritional guidelines for patients with cystic fibrosis (CF) recommend aggressive treatment of growth failure. Oral reduced glutathione (GSH) has been shown to improve cachexia and case reports have demonstrated improved growth in pediatric patients with CF. No controlled studies using oral GSH in CF have, however, been reported. The aim of the study was to determine whether oral GSH could improve growth in CF. Secondarily, to determine whether oral GSH could improve other systemic clinical markers., Methods: We performed a placebo-controlled, randomized, double-blind, repeated-measures clinical trial in 44 pediatric patients with CF ages 18 months to 10 years. Primary outcomes were change in weight percentile, body mass index (BMI) percentile, height percentile, and fecal calprotectin. Secondary outcomes included liver function tests and measures of systemic inflammation. Each participant was studied for 6 months, with data obtained at baseline, 3 months, and 6 months. Blood samples were obtained on the baseline and 6-month visits. Subjects were treated with oral GSH or placebo (calcium citrate), each 65 mg · kg(-1) · day(-1) divided into 3 doses per day at mealtimes, and administered daily for 6 months., Results: The GSH treatment group gained an average of 0.67 standard deviation (SD) in weight-for-age-and sex z score (wfaszs), (19.1 weight percentile points) during the course of 6 months, with no adverse effects (vs placebo with an increase of 0.1 SD in wfaszs [2.1 weight percentile points], P < 0.0001). Fecal calprotectin improved, GSH -52.0 vs placebo 0.5), also BMI for GSH improved 0.69 SD BMI-adjusted-for-age-and-sex z score versus placebo 0.22 SD (BMI percentile 21.7 GSH vs 5.2 placebo), and height 0.2 SD in height-for-age-and-sex z score (hfaszs) GSH versus -0.06 SD hfaszs placebo [height percentile 7.0 GSH vs -2.6 placebo], all P < 0.0001). Secondary outcomes improved significantly, as well., Conclusions: Oral reduced L-GSH significantly improves measures of growth status and gut inflammation in CF.

Objectives: To assess the BMI status of children and adolescents with sickle cell disease (SCD) and determine if zBMI status during adolescence is predicted by gender, childhood zBMI status, disease genotype, and healthcare utilization (emergency department visits or hospitalizations)., Study Design: Medical chart reviews were conducted on 133 patients followed through a regional Comprehensive Sickle Cell Center to obtain anthropometric measures and healthcare utilization data. Gender-specific BMI z-scores were calculated based on Centers for Disease Control (CDC) norms using Epi Info NutStat Software and SPSS generated syntax. Data were summarized categorically across two time periods for each participant: childhood (age 6-12 years) and adolescence (age 13-18 years)., Results: Males were three times more likely to be underweight in adolescence compared to CDC norms, whereas females were three times more likely to be obese in adolescence. In addition, regression analyses indicated that BMI in adolescence was predicted by gender, average weight in childhood, and the average number of emergency department visits., Conclusions: Children with SCD generally exhibit normal growth during childhood and adolescence, although 5-10% are at risk for poor growth or obesity. Prevention and intervention efforts should consider gender, average weight in childhood, and healthcare utilization factors.

Background: Herpes simplex-1 virus (HSV-1) reactivation in the respiratory tract is common in intensive care unit (ICU) patients. However, susceptible ICU populations are poorly defined. Clinical recognition of HSV infection of the respiratory tract is difficult and the impact of such reactivation is not understood., Materials and Methods: A retrospective analysis of HSV-1 positive patients encountered over a 5-year period at a multispecialty ICU was carried out. HSV-1 was identified in respiratory secretions using a qualitative polymerase chain reaction (PCR) technique. Patient charts were reviewed for clinical features that would typify HSV-1 respiratory involvement, and the morbidity and mortality risks found with HSV-1 respiratory involvement., Results: A review of 48 HSV-1 positive ICU patients showed that patients reactivating HSV in the respiratory tract fell into one of the three categories: (1) septic elderly patients with and without ARDS, (2) immunosuppressed patients, especially those receiving high-dose steroids, and (3) post-thoracotomy patients. Abnormalities suggestive of HSV-1 reactivation in the respiratory tract included, haemorrhagic or excessive respiratory secretions, concomitant orofacial herpes (42%), and bronchoscopic abnormalities (hemorrhagic ulcers and mucosal friability) (83%). Twenty eight percent of the HSV-1 infected patients experienced postextubation stridor. HSV-1 reactivation was associated with extended ventilator stays, significant mortality (42%), and ventilator-associated pneumonias (52%)., Conclusions: Identification of susceptible populations and definition of clinical features of HSV-1 related respiratory disease can enable diagnosis of HSV-1 infection in ICU patients. Although detection by a PCR technique can rapidly diagnose HSV-1 reactivation, prospective studies are required to clarify HSV disease versus mere shedding, and understand the impact of HSV-1 reactivation in hospitalized patients.

CFTR mutation, which causes cystic fibrosis (CF), has also recently been identified as causing glutathione system dysfunction and systemic deficiency of reduced glutathione (GSH). Such dysfunction and deficiency regarding GSH may contribute to the pathophysiology of CF. We followed 13 patients (age range 1-27 years) with cystic fibrosis who were using a regimen of reduced glutathione (GSH), including oral glutathione and inhaled buffered glutathione in an uncontrolled, observational study. Dosage ranged from 66-148 mg/kg/day in divided doses, and the term examined was the initial 5.5 months of GSH use (45 days of incrementally adjusted dose, plus 4 months of use at full dosage). Baseline and post-measurements of FEV1 percent predicted, BMI percentile, and weight percentile were noted, in addition to bacterial status and pulmonary exacerbations. Significant improvement in the following clinical parameters was observed: average improvement in FEV1 percent predicted (N=10) was 5.8 percentage points (p<0.0001), average weight percentile (N=13) increased 8.6 points (p<0.001), BMI percentile (N=11) improved on average 1.22 points (p<0.001). All patients improved in FEV1 and BMI, if measured in their case; 12 of 13 patients improved in weight percentile. Positive sputum cultures of bacteria in 11 patients declined from 13 to 5 (p<0.03) with sputum cultures of Pseudomonas aeruginosa becoming negative in 4 of 5 patients previously culturing PA, including two of three patients chronically infected with PA as determined by antibody status. Use of a daily GSH regimen appears to be associated in CF patients with significant improvement in lung function and weight, and a significant decline in bacteria cultured in this uncontrolled study. These findings bear further clinical investigation in larger, randomized, controlled studies.

Objective: To investigate whether the combination of standard medical care (SMC) and short-term cognitive-behavioral family treatment (CBT) in the treatment of recurrent abdominal pain (RAP) was more effective than SMC alone., Methods: Children recently diagnosed with RAP via physician examination were randomized into SMC (n = 29) and SMC plus CBT (n = 40) groups. Outcome measures included multiple dimensions of child and parent reported child pain, somatization, and functional disability, and school absences and physician contacts., Results: Children and parents participating in the combined SMC + CBT intervention reported significantly less child and parent reported child abdominal pain than children in the SMC intervention immediately following the intervention and up to 1 year following study entry, as well as significantly fewer school absences. Significant differences in functional disability and somatization were not revealed., Conclusions: These results, in combination with previous studies, add support to the effectiveness of CBT intervention in reducing the sensory aspects of RAP. Results are discussed with respect to the cost-benefit of integrated medical and short-term psychological services.

Trichloroethylene (TCE) has been identified as a widespread groundwater contaminant. Trichloroacetic acid (TCA) and dichloroacetic acid (DCA) are toxicologically relevant metabolites of TCE that produce tumors in B6C3F1 mice. A sensitive method for measuring these metabolites in plasma has been developed to obtain pharmacokinetic data from TCE exposure. This is particularly important because DCA is more potent at producing hepatoproliferative lesions than TCA. At present, it is unclear whether DCA is produced by humans. Existing gas chromatographic methods cannot detect DCA at low nanogram-per-milliliter levels. A Finnigan TSQ 700 mass spectrometer (MS) with electrospray ionization was used to measure TCA, DCA, and monochloroacetic acid (MCA) in plasma. The MS was operated in negative ion tandem MS mode. The limit of detection for TCA and DCA was 4 ng/ml, and the limit of detection for MCA was 25 ng/mL. Plasma samples from human subjects exposed to 100 ppm TCE for 4 h contained TCA at concentrations as high as 10 micrograms/mL. DCA concentrations were less than 5 ng/mL, and MCA was not detected (less than 25 ng/mL).

Trichloroethylene (TCE) has been identified as an environmental contaminant in groundwater. Trichloroacetic acid (TCA), dichloroacetic acid (DCA), chloral hydrate (CH), trichloroethanol (TCOH), and trichloroethanol glucuronide (TCOG) have been identified as metabolites of TCE. Studies have shown that TCA and DCA are toxicologically significant metabolites that can induce liver tumors in B6C3F1 mice. Methods for the analysis of these metabolites are important for conducting pharmacokinetic studies. In this study, TCA and DCA were derivatized to their methyl esters by dimethyl sulfate under acidic conditions and analyzed by gas chromatography with electron capture detection. In developing a method for esterifying TCA and DCA, the conversion of TCA to DCA was observed in freshly drawn blood upon the addition of acid. After blood was drawn from the animals, the amount of TCA converted to DCA by the addition of acid decreased with time. This conversion could be prevented by freezing blood samples overnight prior to the addition of acid. Further experiments demonstrated that this activity could be restored by the addition of dithionite to inactivate blood samples or the addition of dithionite to methemoglobin prior to the addition of acid. The results reported here show that reduced hemoglobin may be involved in the acid-catalyzed conversion of TCA to DCA.

Culture conditions modulating cell damage from xanthine plus xanthine oxidase-derived partially reduced oxygen species were studied. Porcine thoracic aorta endothelial cells and porcine lung fibroblasts were maintained in monolayer culture. Cells were prelabeled with 51Cr before xanthine plus xanthine oxidase exposure. Endothelial cells showed 30 to 100% more lysis than fibroblasts and thus seemed more sensitive to this oxidant stress. The effect of cell culture age, as indicated by population doubling level (PDL), was examined. Response of low PDL endothelial cells and fibroblasts subjected to oxidant stress was compared with the response of PDL 15 cells. Both low PDL endothelial cells and fibroblasts responded differently to the lytic effect of xanthine oxidase-derived free radicals than did higher PDL cells. Specific activities of the antioxidant enzymes catalase, manganese superoxide dismutase, copper-zinc superoxide dismutase, glutathione peroxidase, and glucose-6-phosphate dehydrogenase were measured in both low and high PDL fibroblasts and endothelial cells. Antioxidant enzyme specific activities could only partially explain the differences in response to oxidant stress between fibroblasts and endothelial cells and between low and high PDL cells. Cell culture medium composition modulated the rate of production, and relative proportions of xanthine plus xanthine oxidase-derived partially reduced species of oxygen, i.e. superoxide, hydrogen peroxide, and hydroxyl radical. Serum content of medium was important in modulating free radical generation; superoxide production rates decreased 32%, H2O2 became undetectable, and hydroxyl radical generation decreased 54% in the presence of 10% serum. The medium protein and iron content also modulated free radical generation. The data suggest that cell culture media constituents, cell type, and cell culture age greatly affect in vitro response of cells subjected to oxidant stress.

We investigated whether familial factors influence the plasma content of sex-steroids and sex-hormone-binding globulin (SHBG) in 98 adult males of 66 families. They had no apparent endocrine dysfunction, The 0800-1100 hr plasma levels of testosterone, 5 alpha-dihydrotestosterone (DHT), estradiol-17 beta (E2) and estrone (E1) were measured by radioimmunoassay. The free fractions of E2 and testosterone were determined by equilibrium dialysis, and binding capacity of SHBG was also calculated. The data were analyzed by analysis of variance. We observed that the differences in the plasma content of testosterone (p = .02). SHBG binding capacity (p = 0.1), and E2 (p = 0.3), free E2 index (p = .05) were all substantially less variable within groups of brothers than among non-brothers. The variability of the plasma concentration of DHT, free testosterone and E1 was not significantly less within brothers than among non-brothers. The correlation between either plasma testosterone content (r = .14) or SHBG binding capacity (r = .12) and percent of ideal body weight was not significant statistically. Age had no effect on the results. Our data suggest that genetic and/or environmental factors may affect the plasma content of testosterone, E2 and SHBG binding capacity.

Utah mortality rates for cerebrovascular disease (ICD numbers 430--438) are 13% below U.S. rates. About 70% of Utahns are members of the Church of Jesus Christ of Latter-day Saints, commonly called Mormons of LDS, which proscribes use of tobacco and alcohol. Other studies on this group have found significantly lower occurrence of many cancers and ischemic heart disease. We tested the hypothesis that Utah's lower cerebrovascular disease (CBVD) mortality was contributed by the LDS population. We classified by religion all CBVD deaths (2,521) (except subarachnoid hemorrhage and cerebral embolism) occurring in the state in 1968--1971. No significant difference was found between LDS and non-LDS, but both groups had mortality rates below U.S. expectation. Although recent studies have reported smoking to be a risk factor for CBVD, we found no consistent difference between the LDS and non-LDS, even in the younger age groups. The results do not support the hypothesis that tobacco is an important etiologic agent in CBVD mortality.

Preparation of the following glycosides is described: 2-aminoethyl beta-D-glycosides of (A) 2-acetamido-3,4,6-trio-O-acetyl-2-deoxy-D-glucopyranose, (B) 2-acetamido-4-O-(2-acetamido-3,4,6-trio-O-acetyl-2-deoxy-beta-D-glucopyranosyl)-3,6-di-O-acetyl-2-deoxy-beta-D-glucopyranose (N,N'-diacetylchitobiose pentaacetate), (C) 4-O-(2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl)-2,3,6-trio-O-acetyl-beta-D-glucopyranose (cellobiose heptaacetate); 8-carboxyoctyl glycosides of (D) cellobiose, and (E) N,N'-diacetylchitobiose. Conjugates were prepared from (A), (B), and (C) by coupling to bovine serum albumin by cyanuric trichloride and subsequent deacetylation; (D) and (E) were coupled to bovine serum albumin by the mixed-anhydride reaction. Conjugates (A) and (B) were insoluble; conjugates (C), (D), and (E) functioned as artificial antigens and gave rise to precipitating antibodies in rabbits. Specificities of the antisera were determined by inhibition studies.

The serological reactivities of polysaccharides isolated from five species of dermatophytes, Microsporum quinckeanum, Trichophyton granulosum, T. interdigitale, T. rubrum, and T. schoenleinii, with rabbit antisera to these species were studied qualitatively by precipitation in gel and quantitatively by complement-fixation analyses. Significant differences in the serological reactivities of the galactomannans I were detected with antisera to T. schoenleinii and T. interdigitale. The differences appeared to be related to the specificity of these antisera for the galactofuranose residues in the polysaccharides. Antisera to M. quinckeanum, T. granulosum, or T. rubrum did not detect differences between the galactomannans I. The serological reactivities of the galactomannans II were different with each of the five antisera. The reactivities of the glucans could be correlated with the amount of alpha 1 --> 6 linked glucopyranose residues when antisera to T. schoenleinii and M. quinckeanum were used.

The contribution of terminal galactofuranose residues to the antigenic specificity and to cross-reactivity of galactomannans isolated from five species of dermatophytes, Microsporum quinckeanum, Trichophyton granulosum, T. interdigitale, T. rubrum, and T. schoenleinii, was investigated. Galactofuranose units were removed from galactomannans I and galactomannans II by mild acid hydrolysis. The resulting mannans were tested for serological reactivity with rabbit antiserum to M. quinckeanum by qualitative precipitation in gel and by quantitative complement-fixation analyses. Our results showed that, with this antiserum, the galactofuranose residues contributed greatly to the antigenic specificity and to cross-reactivity of the galactomannans II, but these residues were less significant as antigenic determinants in the galactomannans I. We have shown that mannans isolated from three Candida species reacted with rabbit antiserum to M. quinckeanum.

Antiserum produced in the rabbit to autoclaved mycelial suspensions of Microsporum quinckeanum reacted with three neutral polysaccharides isolated from each of five species of dermatophytes, M. quinckeanum, Trichophyton granulosum, T. interdigitale, T. rubrum, and T. schönleinii. The serological reactivities of these polysaccharides, grouped as galactomannans I, galactomannans II, and glucans, were compared by qualitative precipitation analyses in gel and quantitative complement-fixation analyses. Significant differences were found among the glucans and galactomannans II but not among the galactomannans I of these species.

The chemical structures and serological specificities of polysaccharides isolated from four species of dermatophytes, Microsporum praecox, Trichophyton ferrugineum, T. sabouraudii, and T. tonsurans, were investigated. Each of these species yielded a mixture of crude polysaccharides which could be separated into three water-soluble, neutral polysaccharides free of nitrogen. These were grouped as galactomannan I, galactomannan II, and glucan. The galactomannans I were quite similar in chemical structure. When measured by complement fixation, their serological cross-reactivities were similar with rabbit antisera to each of these species except T. sabouraudii. The differences in their relative reactivities with this antiserum could be correlated with differences in structure and specificity of this antiserum for galactofuranose end groups. The galactomannans II differed both in chemical structure and in their serological reactivities with antisera to each of these species. The galactomannan II from T. ferrugineum differed most in chemical structure and was the least reactive serologically. The glucans also differed in both structure and serological reactivities.