Sleep Efficiency and Sleep Onset Latency in One Saskatchewan First Nation.

BACKGROUND: Sleep efficiency and sleep onset latency are two measures that can be used to assess sleep quality. Factors that are related to sleep quality include age, sex, sociodemographic factors, and physical and mental health status. This study examines factors related to sleep efficiency and sleep onset latency in one First Nation in Saskatchewan, Canada. METHODS: A baseline survey of the First Nations Sleep Health project was completed between 2018 and 2019 in collaboration with two Cree First Nations. One-night actigraphy evaluations were completed within one of the two First Nations. Objective actigraphy evaluations included sleep efficiency and sleep onset latency. A total of 167 individuals participated, and of these, 156 observations were available for analysis. Statistical analysis was conducted using logistic and linear regression models. RESULTS: More females (61%) than males participated in the actigraphy study, with the mean age being higher for females (39.6 years) than males (35.0 years). The mean sleep efficiency was 83.38%, and the mean sleep onset latency was 20.74 (SD = 27.25) minutes. Age, chronic pain, ever having high blood pressure, and smoking inside the house were associated with an increased risk of poor sleep efficiency in the multiple logistic regression model. Age, chronic pain, ever having anxiety, heart-related illness, and smoking inside the house were associated with longer sleep onset latency in the multiple linear regression model. CONCLUSIONS: Sleep efficiency and sleep onset latency were associated with physical and environmental factors in this First Nation.

Creating a Canadian Indigenous Research Network Against Cancer to Address Indigenous Cancer Disparities.

PURPOSE: In Canada, indigenous peoples' cancer rates have increased, but cancer screening rates tend to be lower. When coupled with poor cancer prognosis, treatment barriers, and inaccessible health care, indigenous patients with cancer experience many unmet needs. Further complicating their journey is a multijurisdictional system that complicates cancer control services, treatments, patient supports, and cancer surveillance. To address these issues, the Canadian Indigenous Research Network Against Cancer (CIRNAC) was developed. This article describes the forerunners and consultative process that created the network and the consensus model developed to ground this network with, by, and for indigenous peoples. METHODS: A consultative workshop was held to (1) establish and increase network membership, (2) enhance partnerships with indigenous communities and other researchers, and (3) develop an indigenous-led research program, new funding, and related initiatives. RESULTS: Participants viewed the CIRNAC as a reflective parallel network led by indigenous peoples that would identify research priorities within Canada, assess how these priorities align with indigenous patients' cancer care and research needs, and cross-check to see if these priorities align with each other. The network would also advocate for indigenous elders/knowledge holders and community grassroot processes to drive research and training, thus demonstrating the power of the community voice and lived experience in research. In addition, the network would foster research partnerships to investigate alternative indigenous models for cancer prevention, care, treatment, and support. CONCLUSION: The CIRNAC evolved as a viable vehicle to address cancer with, for, and by indigenous peoples. The network is guided by a preamble, a set of aims, and an inclusion engagement circle model. It is evolving through major world initiatives, with the aim of formally becoming an internationally linked national network.

Risk Factors for Snoring in Two Canadian First Nations Communities.

Snoring may be an important predictor of sleep-disordered breathing. Factors related to snoring among First Nations people are not well understood in a population with high rates of smoking and excess body weight. An interviewer-administered survey was conducted among 874 individual participants from 406 households in 2012 and 2013 in two Canadian First Nations communities. The survey collected information on demographic variables, individual and contextual determinants of respiratory health and snoring (classified as present versus absent) and self-reported height and weight. Multiple logistic regression analyses were conducted to examine relationships between snoring and potential risk factors adjusting for age and sex. Snoring was present in 46.2% men and 47.0% women. Considering body mass index, 259 people (30.3%) were overweight and 311 (36.4%) were considered obese. The combined current/former smoking rate was 90.2%. Being overweight, obesity, sinus trouble, current smoking status and former smoking were significantly associated with snoring. Exposure to home dampness and mold were suggestive of an association with snoring. To the degree that snoring may be a predictor of possible sleep-disordered breathing, these results indicate that environmental conditions such as smoking and home exposures may be important factors in the pathogenesis of these conditions.

Incidence and prevalence of chronic obstructive pulmonary disease among aboriginal peoples in Alberta, Canada.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a major respiratory disorder, largely caused by smoking that has been linked with large health inequalities worldwide. There are important gaps in our knowledge about how COPD affects Aboriginal peoples. This retrospective cohort study assessed the epidemiology of COPD in a cohort of Aboriginal peoples relative to a non-Aboriginal cohort. METHODS: We used linkage of administrative health databases in Alberta (Canada) from April 1, 2002 to March 31, 2010 to compare the annual prevalence, and the incidence rates of COPD between Aboriginal and non-Aboriginal cohorts aged 35 years and older. Poisson regression models adjusted the analysis for important sociodemographic factors. RESULTS: Compared to a non-Aboriginal cohort, prevalence estimates of COPD from 2002 to 2010 were 2.3 to 2.4 times greater among Registered First Nations peoples, followed by the Inuit (1.86 to 2.10 times higher) and the Métis (1.59 to 1.67 times higher). All Aboriginal peoples had significantly higher COPD incidence rates than the non-Aboriginal group (incidence rate ratio [IRR]: 2.1; 95% confidence interval [CI]: 1.97, 2.27). COPD incidence rates were higher in First Nation peoples (IRR: 2.37; 95% CI: 2.19, 2.56) followed by Inuit (IRR: 1.92; 95% CI: 1.64, 2.25) and Métis (IRR: 1.49; 95% CI: 1.32, 1.69) groups. CONCLUSIONS: We found a high burden of COPD among Aboriginal peoples living in Alberta; a province with the third largest Aboriginal population in Canada. Altogether, the three Aboriginal peoples groups have higher prevalence and incidence of COPD compared to a non-Aboriginal cohort. The condition affects the three Aboriginal groups differently; Registered First Nations and Inuit have the highest burden of COPD. Reasons for these differences should be further explored within a framework of social determinants of health to help designing interventions that effectively influence modifiable COPD risk factors in each of the Aboriginal groups.

Identifying gaps in asthma education, health promotion, and social support for Mi'kmaq families in Unama'ki (Cape Breton), Nova Scotia, Canada.

INTRODUCTION: Asthma is the most common chronic condition affecting Aboriginal youth aged 8 to 12 years in Canada. Research investigating psychosocial challenges associated with asthma is limited. This study examines support resources, support-seeking strategies, support and education needs, and intervention preferences of Aboriginal youth with asthma and their caregivers in an effort to encourage community-wide, health-promoting behaviors. METHODS: We employed a community-based participatory research design to conduct interviews with 21 youths aged 8 to 12 years and 17 caregivers from 5 Mi'kmaq communities in Unama'ki (Cape Breton) Nova Scotia, Canada. After conducting interviews that explored existing and desired social, educational, and health support in participating communities, we held a 2-day asthma camp to engage participants in asthma education, social support networking, and cultural activities. At the camp, we collected data through participant observation, sharing circles, focus groups, and youth drawings of their experiences living with asthma. RESULTS: Our study yielded 4 key findings: 1) asthma triggers included household mold, indoor smoking, pets, season change, strenuous exercise, extreme cold, and humidity; 2) social and educational support is lacking in Mi'kmaq communities despite a strong desire for these services; 3) cultural, linguistic, and geographic barriers to accessing support exist; and 4) family members are primary support resources. CONCLUSION: Improved support and educational resources are needed to foster effective Mi'kmaq asthma support networks. Future asthma interventions for marginalized populations must be culturally meaningful and linguistically accessible to those using and providing asthma support.

Histone H2AX phosphorylation after cell irradiation with UV-B: relationship to cell cycle phase and induction of apoptosis.

Damage to DNA that engenders double-strand breaks (DSBs) triggers phosphorylation of histone H2AX on Ser-139. Expression of phosphorylated H2AX (gammaH2AX) can be revealed immunocytochemically; the intensity of gammaH2AX immunofluorescence (IF) measured by cytometry was reported to correlate with the frequency of DSBs induced by X-ray radiation or by DNA damaging antitumor drugs. The aim of the present study was to measure expression of gammaH2AX following exposure of HeLa and HL-60 cells to a wide range of doses of UV-B light (6.1 J/m(2)-3.45 kJ/m(2)) and using multiparameter flow and laser scanning cytometry (LSC) to correlate DNA damage with cell cycle phase and induction of apoptosis. In both cell lines, the highest degree of H2AX phosphorylation induced by UV was seen in S-phase cells, particularly during early portion of S. In cells that did not replicate DNA (G(1), G(2) and M) the degree of H2AX phosphorylation was markedly lower than that in S-phase cells, and was strongly UV dose-dependent. Furthermore, the level of UV-induced gammaH2AX in G(1), G(2) and M was much higher in HeLa- than in HL-60- cells. Apoptotic cells become apparent >2h after exposure to UV and exhibited nearly an order of magnitude higher intensity of gammaH2AX IF than that initially induced by UV; predominantly S-phase cells underwent apoptosis. While the suppression of DNA replication, by aphidicolin prevented the induction of H2AX phosphorylation by UV in most S phase cells, it had no effect on a small cohort of cells that appeared to be entering S-phase, that expressed very high levels of gammaH2AX. Furthermore, aphidicolin itself induced gammaH2AX in early-S phase cells. The induction of gammaH2AX by UV was inhibited, but the incidence of apoptosis increased, by 5 mM caffeine, a known inhibitor of PI-3-related kinases. The data are consistent with the notion that H2AX phosphorylation observed throughout S phase reflects formation of DSBs due to the collision of replication forks with the UV-induced primary DNA lesions. Induction of gammaH2AX in G(1), G(2) and M is likely a response to the primary DSBs generated during UV exposure and/or DNA repair. It is unclear why the latter process was more pronounced in HeLa than in HL-60 cells.

Adaptation of an amphibian mucociliary clearance model to evaluate early effects of tobacco smoke exposure.

RATIONALE: Inhaled side-stream tobacco smoke brings in all of its harmful components impairing mechanisms that protect the airways and lungs. Chronic respiratory health consequences are a complex multi-step silent process. By the time clinical manifestations require medical attention, several structural and functional changes have already occurred. The respiratory system has to undergo an iterative process of injury, healing and remodeling with every exposure. METHODS: To have a better understanding of the initial changes that take place when first exposed to environmental tobacco smoke, we have developed an exposure model, using the frog palate that closely represents the features of obstructive airways where ciliary dysfunction and mucus hypersecretion occur. RESULTS: Mucus transport was significantly reduced, even after exposure to the smoke of one cigarette (p < 0.05) and even further with 4-cigarettes exposure (p < 0.001). Morphometric and ultrastructural studies by SEM show extensive areas of tissue disruption. Gelatinase zymography shows activation of MMP9 in mucus from palates exposed to tobacco smoke. CONCLUSIONS: The clearance of mucus on the frog palate is significantly reduced after exposure to environmental tobacco smoke. Cilia and the extracellular matrix are anatomically disrupted. Tobacco smoke triggers an increased activity of matrix metalloproteinases associated with a substantial defoliation of ciliated epithelium. These studies enhance the knowledge of the changes in the mucociliary apparatus that occur initially after exposure to environmental tobacco smoke, with the goal of understanding how these changes relate to the genesis of chronic airway pathologies in humans.

Long term azithromycin therapy in cystic fibrosis patients: a study on drug levels and sputum properties.

BACKGROUND: Following reports on the treatment of diffuse panbronchiolitis (DPB), recent studies demonstrate that long term therapy with azithromycin (AZM) is effective in cystic fibrosis (CF) patients. However, the underlying mechanisms remain uncertain. Some macrolides, including AZM, display inhibition of virulence factors and other antipseudomonal effects at subinhibitory levels in vitro. OBJECTIVES: Drug doses used for CF and DPB therapy were investigated to determine whether they achieve corresponding sputum drug levels in CF patients in vivo. METHODS: In an open, prospective study, 14 CF patients with chronic Pseudomonas aeruginosa airway infection received 250 mg AZM either daily ('high dose') or twice weekly ('low dose') for 12 weeks. Viscoelasticity of sputum was assessed by magnetic microrheology. RESULTS: AZM accumulated in sputum by two orders of magnitude over a period of four weeks. In the following steady state, median AZM concentrations in sputum were 9.5 microg/mL (0.6 to 79.3 microg/mL, interquartiles 1.4 to 33.4 microg/mL) and 0.5 microg/mL (range less than 0.1 [below detection level] to 5.2 microg/mL, interquartiles 0.2 to 1.4 microg/mL) in the high and low dose groups, respectively. Viscoelasticity improved in all patients but one. CONCLUSIONS: The findings suggest that antipseudomonal activity has to be considered among the potential mechanisms of macrolide therapy. Further, viscoelasticity may be a valuable parameter in future clinical trials.

Thioredoxin liquefies and decreases the viscoelasticity of cystic fibrosis sputum.

The persistent and viscous nature of airway secretions in cystic fibrosis (CF) disease leads to airway obstruction, opportunistic infection, and deterioration of lung function. Thioredoxin (Trx) is a protein disulfide reductase that catalyzes numerous thiol-dependent cellular reductive processes. To determine whether Trx can alter the rheological properties of mucus, sputum obtained from CF patients was treated with TRX and its reducing system (0.1 microM thioredoxin reductase + 2 mM NADPH), and liquid phase-gel phase ratio (percent liquid phase) was assessed by compaction assay. Exposure to low Trx concentrations (1 microM) caused significant increases in the percentage of liquid phase of sputum. Maximal increases in percent liquid phase occurred with 30 microM Trx. Additional measurements revealed that sputum liquefaction by the Trx reducing system is dependent on NADPH concentration. The relative potency of the Trx reducing system also was compared with other disulfide-reducing agents. In contrast with Trx, glutathione and N-acetylcysteine were ineffective in liquefying sputum when used at concentrations <1 mM. Sputum viscoelasticity, measured by magnetic microrheometry, also was diminished significantly following 20-min treatment with 3, 10, or 30 microM Trx. Similarly, this reduction in viscoelasticty also was dependent on NADPH concentration. Further investigation has indicated that Trx treatment increases the solubility of high-molecular-weight glycoproteins and causes redistribution of extracellular DNA into the liquid phase of sputum. Recognizing that mucins are the major gel-forming glycoproteins in mucus, we suggest that Trx alters sputum rheology by enzymatic reduction of glycoprotein polymers present in sputum.

Regulation of Cl- secretion by alpha2-adrenergic receptors in mouse colonic epithelium.

Previous studies have shown that alpha2 adrenoceptor (alpha2AR) agonists inhibit electrolyte secretion in colonic epithelia, but little is known about the molecular mechanisms involved in this process. In this study we examined the effect of alpha2AR activation on transepithelial anion secretion across isolated murine colonic epithelium. We found that alpha2AR agonists, UK 14,304, clonidine and medetomidine were potent inhibitors of anion secretion, especially in the proximal colon. Short circuit current measurements (Isc) in colonic epithelia from normal and cystic fibrosis (CF) mice showed that alpha2AR agonists inhibited basal cystic fibrosis transmembrane conductance regulator (CFTR)-mediated Cl- secretion but had no effect on CFTR activation by cAMP-dependent phosphorylation. Apical administration of an ionophore, nystatin (90 microg ml-1), was used to investigate the effect of UK 14,304 on basolateral K+ transport. The Na+-K+-ATPase current, measured as ouabain-sensitive current in the absence of ion gradients, was unaltered by pretreatment of the tissue with UK 14,304 (1 microM). In the presence of a basolaterally directed K+ gradient, UK 14,304 significantly reduced nystatin-activated Isc indicating that activation of alpha2ARs inhibits basolateral K+ channels. Studies with selective K+ channel inhibitors and openers showed that alpha2AR agonists inhibited KATP channels that were tonically active in mouse colonic epithelia. RT-PCR and pharmacological studies suggested that these channels could be similar to vascular smooth muscle KATP channels comprising Kir6.1/SUR2B or Kir6.2/SUR2B subunits. Inhibition of anion secretion by alpha2AR agonists required activation of pertussis toxin-sensitive Gi/o proteins, but did not involve classical second messengers, such as cAMP or Ca2+. In summary, alpha2ARs inhibit anion secretion in colonic epithelia by acting on basolateral KATP channels, through a process that does not involve classical second messengers.

Antimycin A-induced apoptosis of HL-60 cells.

BACKGROUND: Previous experiments in our laboratory investigating apoptosis induced in HL-60 cells by camptothecin (CAM) have revealed that the sequence and rapidity of the apoptotic phenomena in an individual cell depend on the proliferative state of that cell when it encounters CAM. The role of mitochondria in HL-60 apoptosis was explored using an inhibitor of oxidative phosphorylation, antimycin A (AMA). METHODS: Changes in cell light scatter, binding of annexin V-fluorescein isothiocyanate (FITC), uptake of propidium iodide (PI), and DNA content after membrane fixation/permeabilization were monitored by flow cytometry. Z-VAD-FMK was used to inhibit caspases. Fluorescence microscopy was used to examine cell morphology. RESULTS: Cells in the G1 phase of the cell cycle were the first to exhibit signs of apoptosis in response to 100 microM AMA and some of these cells disintegrated without exposing to phosphatidylserine (PS). Caspase inhibition prevented fragmentation of DNA, the nucleus, and the cell, but only delayed PS exposure and loss of plasma membrane integrity. CONCLUSIONS: The highly active mitochondria of G1-phase HL-60 cells make them particularly sensitive to AMA. PS exposure and plasma membrane damage are mediated by noncaspase molecules released from mitochondria. We hypothesize that if mitochondria are subjected to a sufficiently severe insult, whether indirectly as a result of extensive CAM-induced DNA damage or directly by the effect of AMA on electron transport, the nature and quantities of the proapoptotic molecules released are such that apoptosis proceeds to the point of cell disintegration before the PS exposure pathway is complete.

Pharmacological approaches to discovery and development of new mucolytic agents.

Airway mucus is the secretory product of the mucous cells; it is a variable mixture of water, mucous glycoproteins, low molecular weight ions, proteins, and lipids, whose physical properties are important for airway defense. The factors that contribute to the physical properties of mucus are complex, and there are a number of pharmacological strategies that can potentially serve to improve the clearability of airway mucus. Novel mucoactive approaches include strategies for mucoregulation--decreasing the abnormal volume of mucus secretion--and medications designed to improve the cough clearability of airway secretions. In vitro results suggest potential benefits from the additive effects of selected combinations of mucoactive medications. Further studies are required to confirm these findings, to perform direct assessments of mucus clearability, and to extend the observations to patients with various types of pulmonary diseases where mucoactive treatments are required.

Effects of caspase inhibition on camptothecin-induced apoptosis of HL-60 cells.

BACKGROUND: During camptothecin (CAM)-induced apoptosis of HL-60 cells, the external exposure of phosphatidylserine (PS) can either precede or follow DNA cleavage. The evidence suggests that cells in S-phase when CAM is added undergo rapid DNA, nuclear, and cellular disintegration before exposing PS on the outside of the plasma membrane, whereas cells moving from G1 into S-phase after CAM is added expose PS before they manifest the other phenomena. This study describes further investigations using the broad spectrum caspase inhibitor Z-VAD-FMK. The cells were cultured for a period long enough to ascertain whether a particular phenomenon was only delayed or was blocked completely. METHODS: Changes in cell light scatter, binding of annexin V-fluorescein isothiocyanate (FITC) to PS, uptake of propidium iodide (PI) as a measure of plasma membrane integrity, and DNA content after membrane fixation/permeabilization were monitored by flow cytometry during 24-h cultures. Fluorescence microscopy was used to examine cell morphology. RESULTS: Caspase inhibition blocked DNA cleavage, breakdown of the nuclear membrane, and formation of apoptotic bodies. It also revealed the existence of a CAM-activated early S-phase checkpoint. Cells arrested in early S-phase preceded the appearance of PS-positive cells. Caspase inhibition delayed both PS exposure and loss of plasma membrane integrity but did not prevent either. CONCLUSIONS: The results support the hypothesis that the sequence of apoptotic phenomena in an individual CAM-treated HL-60 cell depends on the stage of proliferation of that cell when it encounters the CAM. They are also consistent with the hypothesis that caspases are not required for PS exposure or the loss of plasma membrane integrity, but they are involved indirectly in promoting these phenomena.

Additive effect of dornase alfa and Nacystelyn on transportability and viscoelasticity of cystic fibrosis sputum.

OBJECTIVE: To investigate the effect of dornase alfa (DA), Nacystelyn (NAL) and their combination on mucociliary transportability and mucus viscoelasticity of cystic fibrosis (CF) sputum, and to assess whether the combination possesses an additive effect. DESIGN: Determination of transportability in frog palate and viscoelasticity in vitro. SETTING: Research laboratory at a medical centre. PATIENTS: Sputa from 15 patients with CF, chronically infected with Pseudomonas aeruginosa, were studied. INTERVENTIONS: Sputum samples were incubated without any drug solution as a control, and with normal saline, DA, NAL and a mixture of DA and NAL in concentrations approximating those achieved in clinical practice. RESULTS: Normal saline (10% volume) by itself had a small effect on CF sputum transportability with a mean increase of 9%, and on viscoelasticity with a mean of decrease of 0.22 log units, respectively, compared with control (incubation without saline). DA (200 nM) further increased the transportability by a mean of 35% versus saline and decreased viscoelasticity by a mean of 0.30 log units. NAL (100 M) increased the transportability by a mean of 32% and decreased viscoelasticity by a mean of 0.22 log units from the levels achieved with saline. The mixture of DA plus NAL at one-half of the above concentration of each agent produced an additional increase in the transportability, by a mean of 18%, and a further decrease in viscoelasticity, by a mean of 0.25 log units, compared with DA or NAL as a single treatment. CONCLUSIONS: The combination of DA and NAL exhibits an additive effect for both the viscoelasticity and transportability of CF sputum samples. The two agents appear to act well together in breaking down the bonding due to extracellular DNA and mucins. Clinical studies should be undertaken to see whether the additive combination at lower concentration produces the anticipated benefits of improved airway clearance and fewer side effects.