Alcohol Use Predicts Longer But More Fragmented Sleep: A Daily Diary Study of Alcohol, Sleep, and PTSD in Nurses.

BACKGROUND: Due to the demanding nature of their profession, nurses are at risk of experiencing irregular sleep patterns, substance use, and fatigue. Evidence supports a reciprocal relationship between alcohol use and sleep disturbances; however, no research has examined such a link in a sample of nurses. One factor that may further impact the dynamic between alcohol and sleep patterns is posttraumatic stress disorder (PTSD) symptoms. We investigated the daily bidirectional associations between alcohol use and several sleep domains (i.e., self-report and actigraphy-determined sleep), and moderation by baseline PTSD symptom severity. METHOD: Over a 14-day period, 392 nurses (92% female; 78% White) completed sleep diaries and actigraphy to assess alcohol use and sleep patterns. Within-person bidirectional associations between alcohol and sleep were examined using multilevel models, with symptoms of PTSD as a cross-level moderator. RESULTS: Daily alcohol use (i.e., ≥ 1 alcoholic beverage; 25.76%) was associated with shorter self-reported sleep onset latency (b = -4.21, p = .003) but longer self-reported wake after sleep onset (b = 2.36, p = .009). Additionally, days with any alcohol use were associated with longer self-reported sleep duration (b = 15.60, p = .006) and actigraphy-determined sleep duration (b = 10.06, p = .037). No sleep variables were associated with next-day alcohol use. Bidirectional associations between alcohol consumption and sleep were similar regardless of baseline PTSD symptoms. CONCLUSION: Our results suggested that on days when nurses drank alcohol, they experienced longer but also more fragmented sleep.

Examining the Factor Structure and Correlates of Motives to Drink Before Attending a Virtual Social Event During COVID-19 Among University Students.

Background: Many university students pregame or drink before a social event. Pregaming carries some risk due to its link to heavy drinking. During the COVID-19 pandemic, there was limited access to many drinking venues (e.g., bars/clubs). Moreover, universities shifted to a virtual format and imposed restrictions on in-person gatherings resulting in the reliance on virtual platforms for class instruction, meetings, and social events. The pandemic facilitated changes in students' drinking behaviors, stress levels, and how they maintained social contact with others. Thus, it is conceivable that during an academic pandemic year, students may have engaged in the act of drinking before attending a virtual social event. Objectives: In the present study, we examined the factor structures/item loadings of the Pregaming Motives Measure-Virtual (PGMM-V) among students (N = 283; Mage = 21.38; women = 69.3%; White = 45.4%, Hispanic = 40.8%) from seven universities who completed an online questionnaire (Spring/Summer-2021). Items from the original Pregaming Motives Measure (Bachrach et al., 2012) were modified to reflect motives to drink before attending a virtual social event. Results: We found evidence for a 2-factor structure model of the PGMM-V which includes social/enhancement and social ease/stress. Bivariate correlations indicated that social/enhancement and social ease/stress were (a) positively associated with frequency of drinking and alcohol consumption prior to attending virtual social events, and (b) general drinking motives (social/enhancement/coping) that align with these motives. Conclusions: The PGMM-V is a promising instrument that could be used in future research designed to understand students' pregaming behaviors for virtual social events as the use of such platforms are increasingly relied upon for social engagement.

The experiences of people with liver disease of palliative and end-of-life care in the United Kingdom-A systematic literature review and metasynthesis.

BACKGROUND: Liver disease is a growing health concern and a major cause of death. It causes multiple symptoms, including financial, psychological and social issues. To address these challenges, palliative care can support people alongside active treatment, and towards the end of life, but little is known about the care experiences of individuals with liver disease in the United Kingdom. This review aimed to explore the palliative and end-of-life care experiences of people with liver disease in the United Kingdom. METHOD: A systematic review was conducted using a five-stage process and following Preferred Reporting Items for Systematic Reviews and Meta Analyses guidelines. Searches were across Web of Science, Scopus, EBSCO and grey literature until 10 May 2023. The review was registered through International Prospective Register of Systematic Reviews (PROSPERO). NVivo 12.5 was used to facilitate data analysis (systematic review registration: PROSPERO CRD42022382649). RESULTS: Of 6035 papers (excluding duplicates) found from searches, five met the inclusion criteria of primary research related to adults with liver disease receiving palliative and/or end-of-life care in the United Kingdom, published in English. Reflexive thematic analysis of the data was conducted. The themes identified were the experiences of people with liver disease of relating to healthcare professionals, using services, receiving support, and experiences of information and communication. These were connected by an overarching concept of disempowerment versus empowerment, with the notion of person-centred care as an important feature. CONCLUSION: This review has found variations in the care experiences of people with advanced liver disease towards the end of life and an overall lack of access to specialist palliative care services. Where services are designed to be person-centred, experiences are more empowering. Further research is needed but with recognition that it is often unclear when care for people with liver disease is palliative or end-of-life. PATIENT AND PUBLIC CONTRIBUTION: An online public involvement workshop was held on 18 April 2023 through Voice (2023). This included four people with liver disease and four carers to discuss the review findings and to design a qualitative research study to further explore the topic.

The Indirect Effects of Recalled Trauma Severity on Pain Ratings among People with Fibromyalgia: a Moderated Mediation Model.

Fibromyalgia syndrome (FM) is a chronic musculoskeletal condition that is accompanied by hypersensitivity to pain. Researchers have examined factors that affect pain ratings among people with FM, such as trauma, depressive symptoms, and coping; however, collectively, the interrelationships among this set of variables, and their relationships to pain, have not been examined. To better understand these relationships, a moderated-mediation model was used to examine how recalled trauma severity, depressive symptoms, relative emotion-focused coping relate to pain ratings. There were 501 participants who were primarily female, White, and ranged in age from 20 to 84 years. All participants had a physician's diagnosis of FM. The results indicated a significant moderated-mediation. Depressive symptoms significantly mediated the relationship between recalled trauma severity and pain ratings, such that greater trauma severity related to more depressive symptoms which in turn were associated with more pain. The mediation chain was moderated by relative emotion-focused coping (i.e., the proportion of emotion-focused coping compared to problem-focused coping), such that when relative emotion-focused coping was used at higher levels, the relationship between recalled trauma severity and depressive symptoms significantly weakened, reducing the indirect association between recalled trauma severity and pain ratings. The findings from the present study indicate that a treatment approach that includes a trauma-focused therapy such as exposure therapy or Emotional Awareness and Expression Therapy should be tested to determine whether these treatments can reduce the impact of past traumas, improve depressive symptoms, decrease pain ratings, and promote more adaptive coping among people with FM.

M.I.C.A.H. Project HEAL: Sustainability of a Faith-Based Community Health Advisor Training Program in Urban Underserved Communities in the USA.

Faith-based organizations (FBOs) can play an important role in improving health outcomes. Lay community health advisors (CHAs) are integral to these efforts. This paper assesses the sustainability of a CHA training program for congregants in African-American and Latino FBOs and subsequent implementation of educational workshops. The program is unique in that a health care chaplain in an academic medical center was central to the program's development and implementation. Forty-eight CHAs in 11 FBOs were trained to teach workshops on cardiovascular health, mental health, diabetes, and smoking cessation. Two thousand four hundred and forty-four participants attended 70 workshops. This program has the potential to be a model to educate individuals and to address health inequities in underserved communities. Health care chaplains in other medical centers may use this as a model for enhancing community engagement and education.

Frontline healthcare workers' mental distress, top concerns, and assessment on hierarchy of controls in response to COVID-19: a cross-sectional survey study.

BACKGROUND: The existing studies showed that frontline healthcare workers during an epidemic experienced unusual stressors and mental distress which even lasted for years after the crisis. It is important to learn about their concerns early to mitigate the negative impact as well as to evaluate disease control from experiences on the front lines for improving responses to the outbreak. The study aimed to provide insights on how to strengthen public health responses to protect healthcare workers both physically and mentally, and effectively control the disease in light of hierarchy of controls. METHODS: A cross-sectional survey was distributed online via Qualtrics to frontline healthcare workers during the COVID-19 through a university's nursing program and received 267 valid responses from 103 certificated nursing assistants, 125 nurses, and 39 other health professionals. A descriptive data analysis with a Chi-square test at a two-sided 0.05 level of significance was performed on factors that potentially affected mental health of healthcare workers and effectiveness of disease control at workplace in five domains. The themes were summarized on open-ended questions. RESULTS: About 30% of the respondents showed the symptom of depression and needed a further investigation. The influencing factors in five domains were examined. Engineering and administrative controls, as well as PPE were widely used in response to COVID-19. The respondents assessed the state and workplace responses to COVID-19 better than the federal government responses. The workplace responses were considered most effective. Multiple factors with a statistically significant correlation with effectiveness of the disease control at workplace were identified. CONCLUSIONS: The study suggested that timely responses at policy level will be more effective than other measures in early prevention and control of the pandemic, mental distress should be addressed in addition to PPE, and nursing programs should consider providing a situation-specific career coaching or counseling for students. A longitudinal study at a larger scale is warranted to capture the variation of time change with the disease control evolvement and across geographic regions.

The Dual Epidemics of 2020: Nursing Leaders' Reflections in the Context of Whole Person/Whole Systems.

The Greater Boston Nursing Collective, a consortium composed of university nursing deans and chief nursing officers within academic medical centers and specialty hospitals in Boston, Massachusetts, was formed in 2014. Since the group's inception, our mission has been to create and reinforce whole-person/whole-system healing environments to improve the health of all communities. Through our collaboration in navigating the dual epidemics of COVID-19 and structural racism within our respective organizations, and across the United States and the world, we share experiences and lessons learned. Our common mission is clearer than ever: to create safe and joyful work environments, to protect the dignity of those we are privileged to serve, and to generate policies to advance health equity to rectify societal forces that have shaped this dual epidemic. We are humbled by the many who persist despite limited rest and respite, and whose stories, innovations, and leadership we are honored to witness and share. They have defined our generation, just as nurses in earlier crises have done: leading through service to others as our purpose and privilege.

CD73 Promotes Glioblastoma Pathogenesis and Enhances Its Chemoresistance via A2B Adenosine Receptor Signaling.

Glioblastoma (GB) is one of the deadliest brain cancers to afflict humans, and it has a very poor survival rate even with treatment. The extracellular adenosine-generating enzyme CD73 is involved in many cellular functions that can be usurped by tumors, including cell adhesion, proliferation, invasion, and angiogenesis. We set out to determine the role of CD73 in GB pathogenesis. To do this, we established a unique GB mouse model (CD73-FLK) in which we spatially expressed CD73 on endothelial cells in CD73-/- mice. This allowed us to elucidate the mechanism of host CD73 versus GB-expressed CD73 by comparing GB pathogenesis in WT, CD73-/-, and CD73-FLK mice. GB in CD73-/- mice had decreased tumor size, decreased tumor vessel density, and reduced tumor invasiveness compared with GB in WT mice. Interestingly, GBs in CD73-FLK mice were much more invasive and caused complete distortion of the brain morphology. We showed a 20-fold upregulation of A2B AR on GB compared with sham, and its activation induced matrix metalloproteinase-2, which enhanced GB pathogenesis. Inhibition of A2B AR signaling decreased multidrug resistance transporter protein expression, including permeability glycoprotein (P-gp) and multidrug resistance-associated protein 1 (MRP1). Further, we showed that blockade of A2B AR signaling potently increased GB cell death induced by the chemotherapeutic drug temozolomide. Together, these findings suggest that CD73 and A2B AR play a multifaceted role in GB pathogenesis and progression and that targeting the CD73-A2B AR axis can benefit GB patients and inform new approaches for therapy to treat GB patients.SIGNIFICANCE STATEMENT Glioblastoma (GB) is the most devastating primary brain tumor. GB patients' median survival is 16 months even with treatment. It is critical that we develop prophylaxes to advance GB treatment and improve patient survival. CD73-generated adenosine has been implicated in cancer pathogenesis, but its role in GB was not ascertained. Here, we demonstrated that host CD73 plays a prominent role in multiple areas of glioblastoma pathogenesis, including promoting GB growth, its angiogenesis, and its invasiveness. We found a 20-fold increase in A2B adenosine receptor (AR) expression on GB compared with sham, and its inhibition increased GB chemosensitivity to temozolomide. These findings strongly indicate that blockade or inhibition of CD73 and the A2B AR are prime targets for future GB therapy.

Loss of CD73 prevents accumulation of alternatively activated macrophages and the formation of prefibrotic macrophage clusters in irradiated lungs.

While radiotherapy is a mainstay for cancer therapy, pneumonitis and fibrosis constitute dose-limiting side effects of thorax and whole body irradiation. So far, the contribution of immune cells to disease progression is largely unknown. Here we studied the role of ecto-5'-nucelotidase (CD73)/adenosine-induced changes in the myeloid compartment in radiation-induced lung fibrosis. C57BL/6 wild-type or CD73-/- mice received a single dose of whole thorax irradiation (WTI, 15 Gy). Myeloid cells were characterized in flow cytometric, histologic, and immunohistochemical analyses as well as RNA analyses. WTI induced a pronounced reduction of alveolar macrophages in both strains that recovered within 6 wk. Fibrosis development in wild-type mice was associated with a time-dependent deposition of hyaluronic acid (HA) and increased expression of markers for alternative activation on alveolar macrophages. These include the antiinflammatory macrophage mannose receptor and arginase-1. Further, macrophages accumulated in organized clusters and expressed profibrotic mediators at ≥25 wk after irradiation (fibrotic phase). Irradiated CD73-/- mice showed an altered regulation of components of the HA system and no clusters of alternatively activated macrophages. We speculate that accumulation of alternatively activated macrophages in organized clusters represents the origins of fibrotic foci after WTI and is promoted by a cross-talk between HA, CD73/adenosine signaling, and other profibrotic mediators.-De Leve, S., Wirsdörfer, F., Cappuccini, F., Schütze, A., Meyer, A. V., Röck, K., Thompson, L. F., Fischer, J. W., Stuschke, M., Jendrossek, V. Loss of CD73 prevents accumulation of alternatively activated macrophages and the formation of prefibrotic macrophage clusters in irradiated lungs.

Multi-Inhibitory Effects of A2A Adenosine Receptor Signaling on Neutrophil Adhesion Under Flow.

A2A adenosine receptor (A2AAR) signaling negatively regulates inflammatory responses in many disease models, but the detailed mechanisms remain unclear. We used the selective A2AAR agonist, ATL313, to examine how A2AAR signaling affects human and murine neutrophil adhesion under flow. Treating neutrophils with ATL313 inhibited selectin-induced, ß2 integrin-dependent slow rolling and chemokine-induced, ß2 integrin-dependent arrest on ICAM-1. ATL313 inhibited selectin-induced ß2 integrin extension, which supports slow rolling, and chemokine-induced hybrid domain "swing-out," which supports arrest. Furthermore, ATL313 inhibited integrin outside-in signaling as revealed by reduced neutrophil superoxide production and spreading on immobilized anti-ß2 integrin Ab. ATL313 suppressed selectin-triggered activation of Src family kinases (SFKs) and p38 MAPK, chemokine-triggered activation of Ras-related protein 1, and ß2 integrin-triggered activation of SFKs and Vav cytoskeletal regulatory proteins. ATL313 activated protein kinase A and its substrate C-terminal Src kinase, an inhibitor of SFKs. Treating neutrophils with a protein kinase A inhibitor blocked the actions of ATL313. In vivo, ATL313-treated neutrophils rolled faster and arrested much less frequently in postcapillary venules of the murine cremaster muscle after TNF-α challenge. Furthermore, ATL313 markedly suppressed neutrophil migration into the peritoneum challenged with thioglycollate. ATL313 did not affect A2AAR-deficient neutrophils, confirming its specificity. Our findings provide new insights into the anti-inflammatory mechanisms of A2AAR signaling and the potential utility of A2AAR agonists in inflammatory diseases.

Determination of Vitamin K1 in Infant, Pediatric, and Adult Nutritionals by HPLC with Fluorescence Detection: Single-Laboratory Validation, First Action 2015.09.

This normal-phase HPLC method with postcolumn reduction and fluorescence detection allows for the quantitative determination of trans vitamin K1 in infant, pediatric, and adult nutritionals. Vitamin K1 is extracted from products with iso-octane after precipitation of proteins and release of lipids with methanol. Prepared samples are injected onto a silica HPLC column where cis and trans vitamin K1 are separated with an iso-octane-isopropanol mobile phase. The column eluent is mixed with a dilute ethanolic solution of zinc chloride, sodium acetate, and acetic acid, and vitamin K1 is reduced to a fluorescent derivative in a zinc reactor column. The resulting hydroquinone is then detected by fluorescence at an excitation wavelength of 245 nm and an emission wavelength of 440 nm. During a single-laboratory validation of this method, repeatability and intermediate precision ranged from 0.6 to 3.5% RSD and 1.1 to 6.0% RSD, respectively. Mean overspike recoveries ranged from 91.9 to 106%. The method demonstrated good linearity over a standard range of approximately 2-90 µg/L trans vitamin K1 with r2 averaging 0.99995 and average calibration errors of <1%. LOQ and LOD in ready-to-feed nutritionals were estimated to be 0.03 and 0.09 µg/100 g, respectively. The method met AOAC Stakeholder Panel on Infant Formula and Adult Nutritionals Standard Method Performance Requirements® and was approved as a first action method at the 2015 AOAC Mid-Year Meeting.

Determination of Vitamin B12 in Infant, Adult, and Pediatric Formulas by HPLC-UV and Column Switching: Collaborative Study, Final Action 2011.10.

AOAC First Action Method 2011.10, Vitamin B12 in Infant and Pediatric Formulas and Adult Nutritionals, was collaboratively studied. This method uses a pH 4.5 sodium acetate buffer and potassium cyanide at 105°C to extract and convert all biologically active forms of vitamin B12 present to cyanocobalamin; octylsilyl (C8) or C18 SPE cartridges to purify and concentrate cyanocobalamin; a combination of size-exclusion and RPLC to isolate cyanocobalamin; and visible absorbance at 550 nm to detect and quantitate cyanocobalamin in infant, pediatric, and adult nutritionals with vitamin B12 concentrations greater than 0.025 µg/100 g ready-to-feed (RTF) liquid. During this collaborative study, nine to 11 laboratories from eight different countries analyzed blind duplicates of 12 infant, pediatric, and adult nutritional formulas. Per the AOAC Expert Review Panel (ERP) on Stakeholder Panel on Infant Formula and Adult Nutritionals (SPIFAN) Nutrient Methods the method demonstrated acceptable repeatability and reproducibility and met SPIFAN Standard Method Performance Requirements (SMPRs®) for the majority of product matrixes analyzed. Vitamin B12 SPIFAN SMPRs for repeatability were ≤15% RSD at vitamin B12 concentrations of 0.01 µg/100 g RTF liquid and ≤7% RSD at vitamin B12 concentrations of 0.2-5.0 µg/100 g RTF liquid. Vitamin B12 SPIFAN SMPRs for reproducibility were ≤11% RSD in products with vitamin B12 concentrations ranging from 0.3 to 5.0 µg/100 g RTF liquid. During this collaborative study, the RSDr ranged from 2.98 to 9.77%, and the RSDR ranged from 3.54 to 19.5%. During previous single-laboratory validation studies, the method LOQ was estimated to be 0.025 µg/100 g RTF liquid.

Determination of Myo-Inositol in Infant, Pediatric, and Adult Formulas by Liquid Chromatography-Pulsed Amperometric Detection with Column Switching: Collaborative Study, Final Action 2011.18.

AOAC First Action Method 2011.18, Myo-Inositol (Free and Bound as Phosphatidylinositol) in Infant and Pediatric Formulas and Adult Nutritionals, was collaboratively studied. With this method free myo-inositol and phosphatidylinositol bound myo-inositol are extracted using two different sample preparation procedures, separated by ion chromatography using a combination of Dionex Carbo Pac PA1 and MA1 columns with column switching, and detected with pulsed amperometry using a gold electrode. Free myo-inositol is extracted from samples with dilute hydrochloric acid and water. Phosphatidylinositol is extracted from samples with chloroform and separated from other fats with silica SPE cartridges. Myo-inositol is then released from the glycerol backbone with concentrated acetic and hydrochloric acids at 120°C. During this collaborative study, nine laboratories from five different countries analyzed blind duplicates of nine infant and pediatric nutritional formulas for both free and phosphatidylinositol bound myo-inositol, and one additional laboratory only completed the free myo-inositol analyses. The method demonstrated acceptable repeatability and reproducibility and met the AOAC Stakeholder Panel on Infant Formula and Adult Nutritionals (SPIFAN) Standard Method Performance Requirements (SMPRs®) for free myo-inositol plus phosphatidylinositol bound myo-inositol for all the matrixes analyzed. SMPRs for repeatability were ≤5% RSD at myo-inositol concentrations of 2-68 mg/100 g ready-to-feed (RTF) liquid. SMPRs for reproducibility were ≤8% RSD in products with myo-inositol concentrations ranging from 2 to 68 mg/100 g RTF liquid. During this collaborative study, repeatability RSDs ranged from 0.51 to 3.22%, and RSDs ranged from 2.66 to 7.55% for free myo-inositol plus phosphatidylinositol bound myo-inositol.

Deficiency of CD73/ecto-5'-nucleotidase in mice enhances acute graft-versus-host disease.

Extracellular ATP and adenosine have immunoregulatory roles during inflammation. Elevated extracellular ATP is known to exacerbate GVHD, and the pharmacologic activation of the adenosine A2A receptor is protective. However, the role of endogenous adenosine is unknown. We used gene-targeted mice and a pharmacologic inhibitor to test the role of adenosine generated by CD73/ecto-5'-nucleotidase in GVHD. In allogeneic transplants, both donor and recipient CD73 were protective, with recipient CD73 playing the dominant role. CD73 deficiency led to enhanced T-cell expansion and IFN-γ and IL-6 production, and the migratory capacity of Cd73-/- T cells in vitro was increased. However, the number of regulatory T cells and expression of costimulatory molecules on antigen-presenting cells were unchanged. A2A receptor deficiency led to increased numbers of allogeneic T cells, suggesting that signaling through the A2A receptor via CD73-generated adenosine is a significant part of the mechanism by which CD73 limits the severity of GVHD. Pharmacologic blockade of CD73 also enhanced graft-versus-tumor activity. These data have clinical implications, as both the severity of GVHD and the strength of an alloimmune antitumor response could be manipulated by enhancing or blocking CD73 activity or adenosine receptor signaling depending on the clinical indication.

Disengagement coping and sleep problems among trauma-exposed adolescents.

BACKGROUND: The formation of healthy sleep patterns is a critical component of positive adolescent development. Dysregulated sleep habits can put youth at risk for the development of a multitude of inimical outcomes, particularly among those who are exposed to a traumatic event. DESIGN AND METHODS: The present study investigated the links between voluntary disengagement coping (e.g., avoidance, denial, wishful thinking) and sleep outcomes among 86 trauma-exposed and non-exposed adolescents between the ages of 12-17 (Mage = 15.44, SD = 1.51; 41.9% female). RESULTS: The relationship between voluntary disengagement coping and sleep outcomes was significant only among trauma-exposed adolescents, such that greater use of voluntary disengagement strategies was associated with greater sleep disturbances and greater daytime dysfunction. CONCLUSIONS: Targeting disengagement coping may be an important strategy to improve sleep health among trauma-exposed adolescents. Continued efforts in improving the efficacy of trauma-exposed adolescent intervention strategies are needed.

Business as usual? Drinking games participation before and during an academic pandemic (COVID-19) year among university students.

Objective: A drinking game (DG) is a risky social drinking activity that is prevalent among university students and promotes rapid alcohol consumption. We examined university students' DG behaviors before and during the COVID-19 pandemic. Method: Students (N = 368; Mage=21.12; women = 72.6%; Hispanic = 44.7%) from seven universities completed an online survey in 2021 (spring/summer). Results: 57% played DGs in-person before the pandemic and continued to play during the pandemic. These students were less worried about their health/symptoms if they were to contract COVID-19, had lower confidence in wearing a mask properly/socially distancing while under the influence of alcohol, consumed more alcohol during the pandemic, and endorsed higher enhancement drinking motives than students who played DGs before but stopped playing during the pandemic (30%). Conclusions: College health practitioners could pay close attention to students who endorse high enhancement motives as they are susceptible to risky DG play.

Ecto-5'-nucleotidase (CD73) decreases mortality and organ injury in sepsis.

The extracellular concentrations of adenosine are increased during sepsis, and adenosine receptors regulate the host's response to sepsis. In this study, we investigated the role of the adenosine-generating ectoenzyme, ecto-5'-nucleotidase (CD73), in regulating immune and organ function during sepsis. Polymicrobial sepsis was induced by subjecting CD73 knockout (KO) and wild type (WT) mice to cecal ligation and puncture. CD73 KO mice showed increased mortality in comparison with WT mice, which was associated with increased bacterial counts and elevated inflammatory cytokine and chemokine concentrations in the blood and peritoneum. CD73 deficiency promoted lung injury, as indicated by increased myeloperoxidase activity and neutrophil infiltration, and elevated pulmonary cytokine levels. CD73 KO mice had increased apoptosis in the thymus, as evidenced by increased cleavage of caspase-3 and poly(ADP-ribose) polymerase and increased activation of NF-κB. Septic CD73 KO mice had higher blood urea nitrogen levels and increased cytokine levels in the kidney, indicating increased renal dysfunction. The increased kidney injury of CD73 KO mice was associated with augmented activation of p38 MAPK and decreased phosphorylation of Akt. Pharmacological inactivation of CD73 in WT mice using α, ß-methylene ADP augmented cytokine levels in the blood and peritoneal lavage fluid. These findings suggest that CD73-derived adenosine may be beneficial in sepsis.

Vitamin C in infant formula and adult/pediatric nutritional formula by HPLC with UV detection: First Action 2012.21.

This method for the determination of vitamin C by HPLC allows for the separation and quantitation of L-ascorbic acid in infant, pediatric, and adult nutritional products. Liquids, semisolids, and powders ranging from 2 to 1000 mg/kg in their consumable forms were analyzed during the method validation. The method met the standard method performance requirements and was approved by an AOAC Expert Review Panel on Infant Formula and Adult Nutritionals on October 2, 2012. During validation, the overall intermediate precision was 2.1% RSD (triplicate determinations on 7-10 days); the within-day precision, or repeatability, was 1.54% RSD (triplicate determinations). Accuracy, as spike recovery, ranged from 97.0 to 100.9%. The method detection and quantitation limits were determined experimentally to be 0.02 and 0.06 mg/L, respectively, in prepared samples.

Determination of vitamins A and E in infant formula and adult/pediatric nutritional formula by HPLC with UV and fluorescence detection: First Action 2012.09.

The method presented is for quantification of alpha-tocopherol (vitamin E), vitamin E acetate, vitamin A acetate, and vitamin A palmitate in infant formula and adult/pediatric nutritionals. The entire lipid fraction, including vitamins A and E, is extracted from product with iso-octane after products are mixed with methanol, which precipitates proteins and disrupts micelles freeing lipids for extraction. Vitamin A palmitate, vitamin A acetate, and vitamin E acetate are separated from alpha-tocopherol on a 3 cm silica column with a 1% methylene chloride, 0.06% isopropanol in iso-octane mobile phase; eluted onto a 20 cm silica column; and, after a column switch, further separated on the 20 cm column before UV detection at 325 nm (vitamin A palmitate and vitamin A acetate) and 285 nm (vitamin E acetate). Alpha-Tocopherol is further separated from other extraneous compounds on the 3 cm silica column and detected by fluorescence at excitation and emission wavelengths of 295 and 330 nm, respectively. Quantification limits in ready-to-feed products were estimated to be 80 IU/L for vitamin A palmitate, 207 International Units (IU)/L for vitamin A acetate, 2.4 mg/L for vitamin E acetate, and < 0.15 mg/L for alpha-tocopherol. Over-spike recoveries and intermediate precision averaged 100.4 and 2.09% RSD for vitamin A palmitate, 100.4 and 1.52% RSD for vitamin E acetate, and 99.6 and 3.02% RSD for alpha-tocopherol. Vitamin A acetate spike recovery data averaged 96.6%, and the intermediate precision for the only product fortified with vitamin A acetate was 2.75% RSD.

CD73-generated adenosine promotes osteoblast differentiation.

CD731 is a GPI-anchored cell surface protein with ecto-5'-nucleotidase enzyme activity that plays a crucial role in adenosine production. While the roles of adenosine receptors (AR) on osteoblasts and osteoclasts have been unveiled to some extent, the roles of CD73 and CD73-generated adenosine in bone tissue are largely unknown. To address this issue, we first analyzed the bone phenotype of CD73-deficient (cd73(-/-)) mice. The mutant male mice showed osteopenia, with significant decreases of osteoblastic markers. Levels of osteoclastic markers were, however, comparable to those of wild-type mice. A series of in vitro studies revealed that CD73 deficiency resulted in impairment in osteoblast differentiation but not in the number of osteoblast progenitors. In addition, over expression of CD73 on MC3T3-E1 cells resulted in enhanced osteoblastic differentiation. Moreover, MC3T3-E1 cells expressed adenosine A(2A) receptors (A(2A)AR) and A(2B) receptors (A(2B)AR) and expression of these receptors increased with osteoblastic differentiation. Enhanced expression of osteocalcin (OC) and bone sialoprotein (BSP) observed in MC3T3-E1 cells over expressing CD73 were suppressed by treatment with an A(2B)AR antagonist but not with an A(2A) AR antagonist. Collectively, our results indicate that CD73 generated adenosine positively regulates osteoblast differentiation via A(2B)AR signaling.