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Bromazolam is a designer benzodiazepine that was first detected in British Columbia in January 2021. Postmortem cases were analyzed using a comprehensive blood drug screening procedure by liquid chromatography–high-resolution mass spectrometry before being retrospectively analyzed using an in-house novel psychoactive substances data processing method. Bromazolam was detected in 41 postmortem cases in 2021 and quantitatively confirmed by standard addition, using liquid chromatography–tandem mass spectrometry. The mean bromazolam concentration observed was 11.4 ± 53.7 ng/mL (median concentration: 1.6 ng/mL), with a range from 0.5 to 319.3 ng/mL and the majority of cases co-occurring with fentanyl. These low concentrations may be indicative of a presumed enhancement of opioid effects, rather than being used as a stand-alone drug. Bromazolam was always detected with opioids (fentanyl and carfentanil), stimulants (methamphetamine) and/or other benzodiazepines (etizolam and flualprazolam). To our knowledge, this is the first report to provide concentrations of bromazolam in postmortem blood samples in Canada.

Early detection of lung infection is critical to clinical diagnosis, treatment, and monitoring. Measuring volatile organic compounds (VOCs) in exhaled breath has shown promise as a rapid and accurate method of evaluating disease metabolism and phenotype. However, further investigations of the role and function of VOCs in bacterial-host-stress response is required and this can only be realised through representative

Early detection of lung infection is critical to clinical diagnosis, treatment, and monitoring. Measuring volatile organic compounds (VOCs) in exhaled breath has shown promise as a rapid and accurate method of evaluating disease metabolism and phenotype. However, further investigations of the role and function of VOCs in bacterial-host-stress response is required and this can only be realised through representative in vitro models. In this study we sampled VOCs from the headspace of A549 cells at an air-liquid interface (ALI). We hypothesised VOC sampling from ALI cultures could be used to profile potential biomarkers of S. aureus lung infection. VOCs were collected using thin film microextraction (TFME) and were analysed by thermal desorption-gas chromatography-mass spectrometry. After optimising ALI cultures, we observed seven VOCs changed between A549 and media control samples. After infecting cells with S. aureus, supervised principal component-discriminant function analysis revealed 22 VOCs were found to be significantly changed in infected cells compared to uninfected cells (p < 0.05), five of which were also found in parallel axenic S. aureus cultures. We have demonstrated VOCs that could be used to identify S. aureus in ALI cultures, supporting further investigation of VOC analysis as a highly sensitive and specific test for S. aureus lung infection.

BACKGROUND: Low airway surface pH is associated with many airway diseases, impairs antimicrobial host defense, and worsens airway inflammation. Inhaled Optate is designed to safely raise airway surface pH and is well tolerated in humans. Raising intracellular pH partially prevents activation of SARS-CoV-2 in primary normal human airway epithelial (NHAE) cells, decreasing viral replication by several mechanisms. METHODS: We grew primary NHAE cells from healthy subjects, infected them with SARS-CoV-2 (isolate USA-WA1/2020), and used clinical Optate at concentrations used in humans in vivo to determine whether Optate would prevent viral infection and replication. Cells were pretreated with Optate or placebo prior to infection (multiplicity of infection = 1), and viral replication was determined with plaque assay and nucleocapsid (N) protein levels. Healthy human subjects also inhaled Optate as part of a Phase 2a safety trial. RESULTS: Optate almost completely prevented viral replication at each time point between 24 h and 120 h, relative to placebo, on both plaque assay and N protein expression (P < .001). Mechanistically, Optate inhibited expression of major endosomal trafficking genes and raised NHAE intracellular pH. Optate had no effect on NHAE cell viability at any time point. Inhaled Optate was well tolerated in 10 normal subjects, with no change in lung function, vital signs, or oxygenation. CONCLUSIONS: Inhaled Optate may be well suited for a clinical trial in patients with pulmonary SARS-CoV-2 infection. However, it is vitally important for patient safety that formulations designed for inhalation with regard to pH, isotonicity, and osmolality be used. An inhalational treatment that safely prevents SARS-CoV-2 viral replication could be helpful for treating patients with pulmonary SARS-CoV-2 infection.

Although pressurized metered dose inhaler (pMDI) education is a routine part of childhood asthma management and encouraging ‘optimal breathing patterns’ (i.e. slowly, deeply, completely, and with a mouth seal on the mouthpiece) is an integral part of recommended pMDI education, there is currently no quantifiable way to determine if a child is inhaling their medication correctly or optimally through a valved holding chamber (VHC). The TipsHaler™ (tVHC) is a prototype VHC device that measures inspiratory time, flow, and volume without changing the properties of the medication aerosol. The measurements in vivo recorded by the tVHC can be downloaded and transferred to a spontaneous breathing lung model to simulate the inhalational patterns in vitro and also determine the deposition of inhaled aerosol mass with each pattern. We hypothesized that pediatric patients’ inhalational patterns when using a pMDI would improve after active coaching via tVHC. This would increase the pulmonary deposition of inhaled aerosols in an in vitro model. To test this hypothesis, we conducted a single-site, prospective, pilot, pre-and-post intervention study paired with a bedside-to-bench experiment. Healthy, inhaler-naïve subjects used a placebo inhaler in conjunction with the tVHC before and after coaching and recorded inspiratory parameters. These recordings were then implemented into a spontaneous breathing lung model during albuterol MDI delivery, and pulmonary deposition of albuterol was quantified. In this pilot study, active coaching resulted in a statistically significant increase in inspiratory time (n = 8, p= 0.0344, 95%CI: 0.082 to ∞). tVHC recorded inspiratory parameters obtained from patients were successfully implemented in the in vitro model, which demonstrated that both inspiratory time (n = 8, r = 0.78, p < 0.001, 95%CI: 0.47–0.92) and volume (n = 8, r = 0.58, p = 0.0186, 95%CI: 0.15–0.85) strongly correlate with pulmonary deposition of inhaled drugs.

Endothelial hemoglobin (Hb)α regulates endothelial nitric oxide synthase (eNOS) biochemistry. We hypothesized that Hb could also be expressed and biochemically active in the ciliated human airway epithelium. Primary human airway epithelial cells, cultured at air–liquid interface (ALI), were obtained by clinical airway brushings or from explanted lungs. Human airway Hb mRNA data were from publically available databases; or from RT-PCR. Hb proteins were identified by immunoprecipitation, immunoblot, immunohistochemistry, immunofluorescence and liquid chromatography- mass spectrometry. Viral vectors were used to alter Hbβ expression. Heme and nitrogen oxides were measured colorimetrically. Hb mRNA was expressed in human ciliated epithelial cells. Heme proteins (Hbα, β, and δ) were detected in ALI cultures by several methods. Higher levels of airway epithelial Hbβ gene expression were associated with lower FEV1 in asthma. Both Hbβ knockdown and overexpression affected cell morphology. Hbβ and eNOS were apically colocalized. Binding heme with CO decreased extracellular accumulation of nitrogen oxides. Human airway epithelial cells express Hb. Higher levels of Hbβ gene expression were associated with airflow obstruction. Hbβ and eNOS were colocalized in ciliated cells, and heme affected oxidation of the NOS product. Epithelial Hb expression may be relevant to human airways diseases.

Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 infection, has led to a pandemic of acute respiratory illness. Pharmacologic treatments for COVID-19 have included treatments that target infection prevention, prevention of viral replication, reduce inflammation, and manage symptoms of respiratory failure caused by the disease. This is a review of key pharmacologic treatments for COVID-19 based on peer-reviewed articles from 2020.

Despite early diagnosis of cystic fibrosis (CF) through newborn screening, a substantial proportion of infants and young children with CF still demonstrate physiologic and structural evidence of lung disease progression, such as obstructive airway disease and bronchiectasis. The growing availability of highly effective CF transmembrane conductance regulatory modulator therapy to the vast majority of people with CF has led to the potential to alter the natural history of CF lung disease, but to assess the full impact of these therapies on CF lung disease and to help guide treatment, sensitive measures of early and mild disease are needed. Chest imaging using computed tomography or magnetic resonance imaging is one approach, but technologic barriers and/or concern about exposure to ionizing radiation may limit its use. However, advances in physiologic measurement techniques and exhaled breath analysis offer another option for assessment of CF lung disease.

Pneumonia is a significant risk for critically ill, mechanically ventilated (CIMV) patients. Diagnosis of pneumonia generally requires a combination of clinician-guided diagnoses and clinical scoring systems. Exhaled breath condensate (EBC) can be safely collected non-invasively from CIMV patients. Hundreds of biomarkers in EBC are associated with acute disease states, including pneumonia. We evaluated cytokines in EBC from CIMV patients and hypothesized that these biomarkers would correlate with disease severity in pneumonia, sepsis, and death. EBC IL-2 levels were associated with chest radiograph severity scores (odds ratio = 1.68; 95% confidence interval = 1.09–2.60; P = 0.02). EBC TNF-α levels were also associated with pneumonia (odds ratio = 3.20; 95% confidence interval = 1.19–8.65; P = 0.02). The techniques and results from this study may be useful for all mechanically ventilated patients.

BACKGROUND: Intravenous formulations of epoprostenol are frequently delivered via nebulizer to treat pulmonary hypertension in acutely ill patients. Although their efficacy as pulmonary vasodilators has been shown to be comparable to inhaled nitric oxide, the local effects of these formulations within the airways have not been determined. We hypothesized that the alkaline diluents of these compounds would lead to increased airway epithelial cell death and ciliary cessation. METHODS: Human bronchial epithelial cells were exposed to epoprostenol in glycine and arginine diluents or control fluid. Ciliary beat frequency, lactate dehydrogenase, and total RNA levels were measured before and after exposure. Results were compared between exposure and control groups. RESULTS: Ciliary beat frequency ceased immediately after exposure to epoprostenol with both diluents. Lactate dehydrogenase levels increased by 200% after exposure to epoprostenol and glycine diluent (P = .002). Total RNA levels were undetectable after exposure to epoprostenol and arginine, indicating complete cell death and lysis (P = .015). Ciliary beat frequency ceased after 30 s of exposure to epoprostenol and glycine (P = .008). There was no difference between cells exposed to epoprostenol and those exposed only to diluent. CONCLUSIONS: Exposure to intravenous formulations of epoprostenol in glycine and arginine caused increased cell death and ciliary cessation in bronchial epithelial cells. These findings suggest that undesired local effects may occur when these compounds are delivered as inhaled aerosols to patients.

Asthma is an obstructive airway disease affecting children and adults throughout the world. It is a heterogeneous disease with a variety of causes and treatments. Research in the diagnosis, treatment, and management of asthma is ongoing, and there were > 8,000 publications on asthma in 2019. This paper reviews several research articles about asthma from 2019 that are most relevant for practicing respiratory therapists caring for patients with asthma.

Exhaled breath condensate (EBC) is a promising source of biomarkers of lung disease. EBC research and utility has increased substantially over the past 2 decades. This review summarizes many of the factors regarding the composition of EBC, its collection, and analysis for the utility of both clinicians and researchers.

Normal breathing for healthy humans is taken for granted; it occurs without conscious effort using ambient (1-atmosphere) pressure with 21% oxygen (O(2)) concentration. The body automatically adjusts for stress, exercise, altitude, and mild disease by increasing the volume and frequency of breathing. Longer term adaptations for exercise and altitude include increases in red blood cell counts and higher concentrations of capillaries in muscle tissue. When more challenging external environmental conditions or pulmonary illnesses exceed the capability for these adaptations, the human system requires technology to maintain sufficient ventilation to preserve life. On the environmental side there are two conditions to be addressed: toxicity of the surrounding atmosphere and changes in external pressure and O(2) concentration. On the medical side, mechanisms for assisting breathing include O(2) supplementation at ambient pressure, positive pressure/flow without additional O(2), or a combination of both. This overview describes the various technologies applied to maintaining a safe breathing environment. Topics for environmental intervention include filter-based and flowing air-supply masks for toxic environments (occupational and laboratory protection), and on-demand gas supply systems for firefighters, self-contained underwater breathing apparatus (SCUBA) divers, and altitude (high performance aircraft, spacecraft) applications. The topics for medical intervention include nasal cannula, continuous positive airway pressure (CPAP), and medical ventilators. The primary purpose of this article is to provide a basic understanding of normal human breathing and the adaptation of breathing in different environments using available technologies.

Pulmonary hypertension is a life-threatening condition that affects people of all ages that can occur as an idiopathic disorder at birth or as part of a variety of cardiovascular and infectious disorders. It is commonly treated with inhaled pulmonary vasodilators such as nitric oxide and less frequently using formulations and analogs of prostacyclin. To minimize systemic effects and preserve pulmonary vasodilation, vasodilators are often administered directly into the airway. Nitric oxide is the only USA Food and Drug Administration-approved inhaled pulmonary vasodilator that can be used during mechanical ventilation. Over the past two decades, interest has grown in the use of aerosolized prostacyclin and prostacyclin analogs for the treatment of pulmonary hypertension during mechanical ventilation. Clinicians who administer inhaled prostacyclin may not have a clear understanding of its risks because of the lack of data from large clinical trials examining safety and efficacy; moreover, its safe use remains poorly documented. The off-label use of drugs is legitimate, but prescribers must recognize the potential complications and liability in doing so. This manuscript aims to address potential problems related to the aerosol administration of pulmonary vasodilators in the mechanically ventilated neonatal patient.

Exhaled breath comprises gas-phase and aerosolized fractions of water vapor and particles. The condensed water vapor fraction is referred to as exhaled breath condensate (EBC), whereas the aerosolized particle fraction, when collected separately, is termed exhaled breath aerosols (EBA). Both of these fractions contain biomarkers relevant to pulmonary and systemic physiology. EBC is easy and safe to collect, and sampling methods have been standardized for nearly all age groups and disease states, yet the composition of EBC varies between collection method and patient population and therefore care must be taken to ensure sample integrity. This chapter provides an overview of EBC and aerosols, principles of their collection, and validated biomarkers within these matrices.

Mechanical ventilation is frequently used in pediatric patients to ensure adequate gas exchange, ameliorate respiratory distress, and enable resolution of pulmonary or other disorders. However, a number of important challenges remain in the pediatric population because there is a paucity of large-scale randomized controlled trials to generate data and inform clinical practice. This review summarizes a number of discoveries and advances that have been made in pediatric mechanical ventilation from June 2017 to December 2018.

The mechanisms by which transepithelial pressure changes observed during exercise and airway clearance can benefit lung health are challenging to study. Here, we have studied 117 mature, fully ciliated airway epithelial cell filters grown at air-liquid interface grown from 10 cystic fibrosis (CF) and 19 control subjects. These were exposed to cyclic increases in apical air pressure of 15 cmH2O for varying times. We measured the effect on proteins relevant to lung health, with a focus on the CF transmembrane regulator (CFTR). Immunoflourescence and immunoblot data were concordant in demonstrating that air pressure increased F508Del CFTR expression and maturation. This effect was in part dependent on the presence of cilia, on Ca2+ influx, and on formation of nitrogen oxides. These data provide a mechanosensory mechanism by which changes in luminal air pressure, like those observed during exercise and airway clearance, can affect epithelial protein expression and benefit patients with diseases of the airways.

A number of clinical scoring systems for asthma management have been described in the literature over the last four decades.[1][1] These systems are largely used to index severity of disease, quantify changes in disease status, and predict outcomes. Resulting scores can then be used to triage

Exhaled breath condensate (EBC) is a promising source of biomarkers of lung disease. EBC research and utility has increased substantially over the past 2 decades. This review summarizes many of the factors regarding the composition of EBC, its collection, and analysis for the utility of both clinicians and researchers.

BACKGROUND: Tissue factor (TF) canonically functions as the initiator of the coagulation cascade. TF levels are increased in inflamed airways and seem to be important for tumor growth and metastasis. We hypothesized that airway epithelia release TF as part of a wound repair program. OBJECTIVES: The goal of this study was to evaluate whether airway epithelia release TF in response to pro-inflammatory stimuli and to investigate roles of TF in cell growth and repair. METHODS: Airway epithelial cells were exposed to 10 μg/mL of lipopolysaccharide or 1 ng/mL of transforming growth factor β (TGF-β). TF and TGF-β messenger RNA and protein were measured in cell lysate and culture media, respectively. Signaling pathways were evaluated by using selective agonists and inhibitors. Airway epithelia were mechanically injured in the presence of TF and tissue factor pathway inhibitor to investigate their roles in wound repair. RESULTS: TF protein levels increased in cell media after exposure to lipopolysaccharide (P < .01) but only in growing cells, and this action was blocked when exposed to an extracellular signal-regulated kinase inhibitor or a “small” worm phenotype and mothers against Decapentaplegic inhibitor. TF protein also increased in the presence of TGF-β (P < .05). Exposure to TF pathway inhibitor decreased the rate of cell growth by 60% (P < .05), and exposure to TF increased the rate of airway healing after injury by 19% (P < .05). CONCLUSIONS: Growing airway epithelia release TF when exposed to lipopolysaccharide or TGF-β. TF reduces wound-healing time in airway epithelia and therefore may be important to airway recovery after injury.

Exhaled breath is a robust matrix of biomarkers divided between three fractions – gaseous breath, volatile breath, and breath condensate. Breath is collected non-invasively through bags (for gaseous breath), cold condensation chambers (breath condensate), and adsorbent traps (volatile breath). Due to the incredibly dilute nature of breath matrices, breath biomarker analysis requires precise analytical techniques, highly sensitive technology and often challenges the limit of detection of even the most advanced assays. Interest and advances in breath collection, analysis, and use have increased in recent years largely due to advances in analytical technology. Approved and validated breath tests are available as tools for researchers and clinicians. Novel development is ongoing. This article reviews the current applications for exhaled breath biomarkers.

To better understand the impact of hypoxia and hyperoxia on neonatal morbidity and mortality, we examined the number of hypoxia and hyperoxia events as well as percentage of time spent outside oxygen saturation targets in relationship to threshold retinopathy of prematurity (tROP) and mortality in preterm infants.Saturation data in 2-second sampling from pulse oximeters was prospectively collected in a single NICU. Average SaO2, low and high saturation events, duration of events, and percentage of time infants spent outside of oxygen saturation range were collected and analyzed continuously during the hospitalization.102 infants1500g or32 weeks gestation were enrolled. There were 125, 112, and 43 hypoxia events/day and 106, 80, and 34 hyperoxia events/day for tROP (N=8), non-survivor (N=16) and non-tROP patients (N=78), respectively. Infants were outside saturation targets for 2:35, 1:38, and 1:03 (hypoxia) and 2:02, 1:25, and 0:38 hours/day (hyperoxia) for tROP, non-survivor and non-tROP, respectively. Time spent outside saturation range (hypoxia, hyperoxia and total time) for the hospital course was higher in tROP (P≤0.006) and non-survivor (P≤0.005) compared with non-tROP patients. The three groups defined themselves in the first 10 days after birth, with regard to duration of hypoxia (P=0.0003), hyperoxia (P=0.0004) and total time outside the targeted saturation range (P=0.0006).Information such as the duration and number of hypoxia and hyperoxia events, as well as total time outside the targeted saturation range, could be factored into assessing clinical interventions and research studies in the prevention, treatment and evaluation of neonatal outcomes.

Background In a patient with positive serum serology for coccidioidomycosis, the differential diagnosis of concurrent pleural effusions can be challenging. We, therefore, sought to clarify the performance characteristics of biochemical, serologic, and nucleic-acid-based testing in an attempt to avoid invasive procedures. The utility of adenosine deaminase (ADA), coccidioidal serology, and polymerase chain reaction (PCR) in the evaluation of pleuropulmonary coccidioidomycosis has not been previously reported. Methods Forty consecutive patients evaluated for pleuropulmonary coccidioidomycosis were included. Demographic data, pleural fluid values, culture results, and clinical diagnoses were obtained from patient chart review. ADA testing was performed by ARUP Laboratories, coccidioidal serologic testing was performed by the University of California-Davis coccidioidomycosis serology laboratory, and PCR testing was performed by the Translational Genomics Research Institute using a previously published methodology. Results Fifteen patients were diagnosed with pleuropulmonary coccidioidomycosis by European Organization for the Research and Treatment of Cancer/Mycoses Study Group criteria. Pleural fluid ADA concentrations were Conclusion Contrary to prior speculation, ADA levels in pleuropulmonary coccidioidomycosis were not elevated in this study. The sensitivity and specificity of coccidioidal serologic testing in nonserum samples remained high, but the clinical usefulness of PCR testing in pleural fluid was disappointing and was comparable to pleural fluid culture.

Changes in cerebral blood flow (CBF) during a hyperglycemic challenge were mapped, using perfusion-weighted MRI, in a group of non-human primates. Seven female baboons were fasted for 16 h prior to 1-h imaging experiment, performed under general anesthesia, that consisted of a 20-min baseline, followed by a bolus infusion of glucose (500 mg/kg). CBF maps were collected every 7 s and blood glucose and insulin levels were sampled at regular intervals. Blood glucose levels rose from 51.3 ± 10.9 to 203.9 ± 38.9 mg/dL and declined to 133.4 ± 22.0 mg/dL, at the end of the experiment. Regional CBF changes consisted of four clusters: cerebral cortex, thalamus, hypothalamus, and mesencephalon. Increases in the hypothalamic blood flow occurred concurrently with the regulatory response to systemic glucose change, whereas CBF declined for other clusters. The return to baseline of hypothalamic blood flow was observed while CBF was still increasing in other brain regions. The spatial pattern of extra-hypothalamic CBF changes was correlated with the patterns of several cerebral networks including the default mode network. These findings suggest that hypothalamic blood flow response to systemic glucose levels can potentially be explained by regulatory activity. The response of extra-hypothalamic clusters followed a different time course and its spatial pattern resembled that of the default-mode network.

Structural imaging research offers excellent translational benefits when non-human primate (NHP) models are employed. In this paper, we will discuss the development of anatomical MR imaging protocols for two important applications of structural imaging in NHPs: studies of genetic variability in brain morphology and longitudinal imaging of fetal brain maturation trends. In contrast with imaging studies of adult humans, structural imaging in the NHPs is challenging due to a comparatively small brain size (2- to 200-fold smaller volume, depending on the species). This difference in size is further accentuated in NHP studies of brain development in which fetal brain volumes are 10-50% of their adult size. The sizes of cortical gyri and sulci scale allometrically with brain size. Thus, achieving spatial sampling that is comparable to that of high-quality human studies (approximately 1.0 mm(3)) requires a brain-size-adjusted reduction in the sampling volumes of from 500-to-150 microm(3). Imaging at this spatial resolution while maintaining sufficient contrast and signal to noise ratio necessitates the development of specialized MRI protocols. Here we discuss our strategy to optimize the protocol parameters for two commonly available structural imaging sequences: MPRAGE and TrueFisp. In addition, computational tools developed for the analysis of human structural images were applied to the NHP studies. These included removal of non-brain tissues, correction for RF inhomogeneity, spatial normalization, building of optimized target brain and analysis of cerebral gyrification and individual cortical variability. Finally, recent findings in the genetics of cerebral gyrification and tracking of maturation trends in the fetal, newborn and adult brain are described.

In the search for bioactive sphingosine 1-phosphate (S1P) receptor ligands, a series of 2-amino-2-heterocyclic-propanols were synthesized. These molecules were discovered to be substrates of human-sphingosine kinases 1 and 2 (SPHK1 and SPHK2). When phosphorylated, the resultant phosphates showed varied activities at the five sphingosine-1-phosphate (S1P) receptors (S1P(1-5)). Agonism at S1P(1) was displayed in vivo by induction of lymphopenia. A stereochemical preference of the quaternary carbon was crucial for phosphorylation by the kinases and alters binding affinities at the S1P receptors. Oxazole and oxadiazole compounds are superior kinase substrates to FTY720, the prototypical prodrug immunomodulator, fingolimod (FTY720). The oxazole-derived structure was the most active for human SPHK2. Imidazole analogues were less active substrates for SPHKs, but more potent and selective agonists of the S1P(1) receptor; additionally, the imidazole class of compounds rendered mice lymphopenic.

Purpose: Development of prostate cancer prevention strategies is an important priority to overcome high incidence, morbidity, and mortality. Recently, we showed that Nexrutine, an herbal extract, inhibits prostate cancer cell proliferation through modulation of Akt and cAMP-responsive element binding protein (CREB)–mediated signaling pathways. However, it is unknown if Nexrutine can be developed as a dietary supplement for the prevention of prostate cancer. In this study, we used the transgenic adenocarcinoma of mouse prostate (TRAMP) model to examine the ability of Nexrutine to protect TRAMP mice from developing prostate cancer. Experimental Design: Eight-week-old TRAMP mice were fed with pelleted diet containing 300 and 600 mg/kg Nexrutine for 20 weeks. Efficacy of Nexrutine was evaluated by magnetic resonance imaging at 18 and 28 weeks of progression and histologic analysis of prostate tumor or tissue at the termination of the experiment. Tumor tissue was analyzed for modulation of various signaling molecules. Results: We show that Nexrutine significantly suppressed palpable tumors and progression of cancer in the TRAMP model. Expression of total and phosphorylated Akt, CREB, and cyclin D1 was significantly reduced in prostate tissue from Nexrutine intervention group compared with tumors from control animals. Nexrutine also inhibited cyclin D1 transcriptional activity in androgen-independent PC-3 cells. Overexpression of kinase dead Akt mutant or phosphorylation-defective CREB inhibited cyclin D1 transcriptional activity. Conclusions: The current study shows that Nexrutine-mediated targeting of Akt/CREB–induced activation of cyclin D1 prevents the progression of prostate cancer. Expression of CREB and phosphorylated CREB increased in human prostate tumors compared with normal tissue, suggesting their potential use as prognostic markers.

The synthesis of N-arylamide phosphonates and related arylether and arylamine analogues provided potent, subtype-selective agonists and antagonists of the five known sphingosine 1-phosphate (S1P) receptors (S1P1–5). To this end, the syntheses of phosphoserine mimetics—selectively protected and optically active phosphonoserines—are described. In vitro binding assays showed that the implementation of phosphonates as phosphate mimetics provided compounds with similar receptor binding affinities as compared to their phosphate precursors. meta-substituted arylamide phosphonates were discovered to be antagonists of the S1P1 and S1P3 receptors. When administered to mice, an antagonist blocked the lymphopenia evoked by a S1P receptor agonist and caused capillary leakage in both lung and kidney.

Phosphothioates may provide metabolic stability when compared to their phosphate counterparts, while retaining the potency and efficacy as agonists at sphingosine-1-phosphate (S1P) G-protein coupled receptors. Unlike their phosphate precursors, phosphothioate compounds with S1P-receptor profiles similar to that of FTY720, an emerging immunomodulator, were shown to evoke prolonged lymphopenia in vivo. Analysis of mouse plasma concentrations for a series of related alcohol/phosphate/phosphothioate compounds showed the conversion of the phosphate to alcohol. These preliminary data highlight the importance of metabolic regulation of S1P receptor ligands.

The purpose of this study was to identify and categorize the variables that clients consider important for forming and strengthening a positive therapeutic alliance. Forty participants were interviewed using the critical incident technique and asked to describe observable behaviors and verbalization

The 25th anniversary of the National Association of Early Childhood Teacher Educators (NAECTE) is a time to celebrate the many successes of an organization dedicated to the preparation of high‐quality teachers of young children. Members share a concern for the importance of early childhood education and a belief that quality teachers of young children are not born but are the result of preparation programs that reflect both historical precedent and current research. Current policy related to fast‐track routes for licensing and high‐stakes testing threaten to undermine accepted practices, values and beliefs that have been the foundation of early childhood teacher preparation. The article which follows is a call to employ the collective strength of NAECTE to advocate for programs that will result in the best teachers for young children.

(1999). Social Reconstructionism for Urban Students. The Clearing House: A Journal of Educational Strategies, Issues and Ideas: Vol. 72, No. 5, pp. 291-294.

Extracellular nucleotides, e.g., ATP, ADP, and UTP, are important signaling molecules which elicit various physiological responses in different tissues. Their degradation is catalyzed by ectonucleotidases which are located on cell surfaces. Most tissues have a mixed population of ectonucleotidases. In this report, the ATP and ADP hydrolyzing ectonucleotidases of chicken gizzard smooth muscle and liver plasma membranes were studied. The two membranes exhibited marked differences in the ratio of ATPase/ADPase activities, activation by divalent cations, thermal stability, responses to detergents and cross-linking agents, and sensitivity to several enzyme inhibitors. The ATPase activity of chicken gizzard membranes is (i) labile to heat and detergents; (ii) activated by concanavalin A and disuccinimidyl suberate, both cross-linking agents; (iii) inhibited by mercurials; and (iv) insensitive to high concentrations of azide, a known inhibitor of ecto-ATP diphosphohydrolases (ecto-ATP/Dase). In contrast, the liver membrane ATPase and ADPase activities are more stable to treatment by heat and detergents and insensitive to cross-linking agents and mercurials, but are inhibited by azide. A low ADP hydrolase activity in the gizzard membranes could be distinguished from both the gizzard ATPase and the liver ATPase/ADPase. This ADP hydrolase, which is markedly stimulated by NBD-Cl, accounts for most of the ADP hydrolysis activity in gizzard membranes. It is concluded that the major ectonucleotidase in the gizzard membranes is an ecto-ATPase whereas that in the liver membranes is an ecto-ATP/Dase. That both membranes contain a mixed population of the ecto-ATPase and ecto-ATP/Dase, but in different proportions, is further demonstrated by immunochemical characterization. The different composition of ectonucleotidases in the two membranes is expected to have an important effect on the regulation of hydrolysis of extracellular ATP as well as the concentration of extracellular adenine nucleotides in the gizzard and liver tissues.

“I'm lost,” he sighed to himself. Then, just ahead he noticed the little postal truck idling under a timeless elm. A woman in a blue toboggan—all bundled up against the cold—hustled back to her truck. She'd just made a delivery. Pulling alongside, he rolled down the passenger's window. “Ma'am,” he said, getting her attention. “Can you tell me how to get to Gratitude Street?” She brightened. “Sure. You got a pen?” When he was ready, she spoke, pointing ahead. “So, at the end of this street, go left. That's Family. You'll know Family because every house is wildly different. You'd swear each one had a different Builder. But actually, the same Builder designed every house. It looks like a mess: there's a treehouse, an igloo, a tent, and a Victorian. Every house and every person on Family look so different they seem out of place. But here's the amazing surprise: despite all the differences, every house has some small elements exactly—and I mean exactly—the same as all the others. “Here's how they discovered it,” she said, warming up. “A long time ago, some angry owners—the homeowners association—drew up rules to make the houses look more alike. Some wanted the igloo removed. Move the teepee, said others. A group felt the treehouse was a terrible eyesore. Then, one day the igloo's heater just died. The Inuit went door to door: Do you have a part that looks like this? she asked. Before you know it, everyone on the street was in an uproar! Everyone's house had exactly the same heater! Exactly! House, teepee, igloo, tent: Whatever your house, it had that exact same heater. As they investigated their homes, they noticed other things were also exact copies. They realized that even with their differences, many things were the same. Nowadays, tourist vans go up and down Family, telling how two houses can be very different but in some ways are still just like each other. The Igloo's Heater is their favorite story. “It's a remarkable place,” she said, wistfully. She refocused. “Sorry, I do get off track. Back to your directions. Go 10 blocks on Family and then turn left onto Can't Do It Alone. Pay attention here or you'll get stuck. Whatever the time of day, people are doing things for others. Somebody's always on a ladder washing a neighbor's windows, or baking a shut-in a loaf of bread or chopping them firewood. Everyone is always helping somebody. Funniest thing, too. Once you're on Can't Do It Alone, you may end up staying! “Take Can't Do It Alone 1 mile. You turn right onto Unexpected just after the railroad tracks. Now, Unexpected seems kind of scary. Some houses are terribly beat up. You'll see folks just hanging out, lurking. Shady characters, you'd think. Lock your doors and windows, be respectful, and have courage. But, the thing about Unexpected is that just when you think the folks around there are all one thing, they aren't! “One time, a fella's car broke down on Unexpected in the worst place possible. He saw a man walking toward him. The man wore a big, long coat, the hood pulled up over his head. The traveler's heart started racing. The figure kept coming, determined. The stranded fella was filled with dread. Then, an insistent knock on his window. Then another, unrelenting. Finally, summoning all his courage, he barely cracked his window, hoping this crazy person couldn't get in.” The postal lady paused dramatically. “The hood came off the top of a head—just a little—revealing a grizzled Old Man. He spoke in a gruff but reassuring voice, ‘Stay out here any longer and you're going to catch your death of cold. Come on in, let's get you warmed up and on your way.’” Now, remember that. When you cross the tracks, Unexpected is just beyond. Unexpected can be scary: unfamiliar and unknown things usually are. But, trust me, the folks and experiences on Unexpected are often just what you need, just when you need it.” “After Unexpected, go about 13 blocks. Life is on your right. Turn there. Now, sometimes you think this Life ain't going to ever end. So far, this trip's had lots of surprises: so many twists and turns you feel utterly lost. Now, though, it seems every house is exactly the same. Same trees. Same hedges. Same roofs. You think, ‘Get me out of here! I don't want this. It's the same thing all the time.’ Will I ever get where I'm going? you wonder. I'm bored. You gotta look harder when you think like that. Trust me, I know. There are lots of little differences all along the way, small reminders to look and listen. Seriously. Keep your eyes open. Look real hard. Don't focus on the sameness. Anticipate surprise. Train your eyes to spot details you usually ignore. Believe me, that's the only way to get through Life. Don't give up, either. As soon as you think, That's it. I'm done. I can't take it anymore, you come up to a big Hill. It's really tough to get up that Hill. Just keep going. Eventually, at the very top, there's a Park. “When you get to the Park, take a break. Look around. It's the highest point in this here part of the world. It's so high, it's hard to catch your breath. But, let your senses steep in the beauty of your surroundings. The air is clear and crisp. The only sounds are birds, leaves rustling, and tree branches soothing one another. Soak it in! Trust me, every day after that, you're going to try to remember just how it felt. So, take your time. Make a picture in your mind. “Oh,” she paused. “When you make it up the Hill, it's tradition to leave a little reminder you were there. Maybe it's a special rock in a hidden place. Or, a paper tucked into a tree's hollow. Or, an initial in the dirt or sand. If you have time, do something like that, okay? From there on, you coast gently downhill until you intersect with the most beautiful spot there ever was. If you've followed my directions, you can't miss it. Right there is Gratitude.” With that, she bid farewell. “More holiday deliveries,” she said. What happened to the Lost Man who asked for directions? No one's quite certain. Once you've started following these directions, it's for sure you'll end up at Gratitude. But, it's unknown how many stops you'll make along the way.