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Inhaled nitric oxide improves survival rates during hypoxia in a sickle cell (SAD) mouse model.

BACKGROUND: The hallmark of sickle cell disease (SCD) is erythrocyte sickling during deoxygenation of the abnormal hemoglobin S (HbS). When HbS is deoxygenated, it aggregates into polymers, resulting in distortion of the erythrocyte structure, producing microvascular thrombosis and ischemia. The transgenic SAD mouse produces three types of human hemoglobin: S, Antilles, and D-Punjab (HbSAD) and provides an animal model for SCD. We studied the effects of nitric oxide (NO) breathing at various doses and time regimens in the presence of severe hypoxia (6% oxygen) using the SAD mouse model. METHODS: Age- and sex-matched control and SAD mice were exposed to 6% oxygen breathing in an environmental chamber and assessed for survival up to 1 h. Animals received different inhaled NO concentrations before and/or during hypoxia. Blood was obtained to evaluate the oxyhemoglobin dissociation curve and measure methemoglobinemia. RESULTS: Pretreatment by breathing NO at 20 ppm by volume in air for 30 min, and continuing to breathe 20 ppm NO during hypoxia resulted in improvement in survival rates in the SAD mouse (75%, n = 8) as compared with control SAD mice (11%, n = 9; P < 0.001). Pretreatment alone or breathing lower doses of NO were not protective. Changes in HbSAD oxygen affinity were not detected with NO breathing, and methemoglobin levels were low in all surviving mice. CONCLUSIONS: Breathing NO produced a rapid, protective effect to severe hypoxic stress in SAD mice. There appears to be a required loading period between NO breathing and its beneficial effect during hypoxic stress, possibly because of the total amount of NO delivered to SAD hemoglobin, blood cell components, and endothelium. NO breathing may be beneficial as a therapeutic intervention in SCD.

Administration, Inhalation↗

The DOF protein, SAD, interacts with GAMYB in plant nuclei and activates transcription of endosperm-specific genes during barley seed development.

The DOF protein, SAD, previously shown to be a transcriptional activator in barley aleurone cells upon seed germination, also has an important role in gene regulation during endosperm development. mRNA was detected in early (10 days after flowering) developing barley seeds where it accumulated in the starchy endosperm, aleurone cells, nucellar projection, vascular tissues and the immature embryo, as shown by RT-PCR and in situ hybridization analyses. The SAD protein, expressed in bacteria, binds to oligonucleotides containing the prolamine box, 5'-A/TAAAG-3'sequence, derived from the promoter regions of the endosperm-specific genes Hor2 and Itr1, encoding a B-hordein and trypsin-inhibitor BTI-CMe, respectively. SAD competed for the same binding sites with another endosperm-expressed DOF protein, BPBF. Transient expression experiments in co-bombarded developing endosperms demonstrated that SAD trans-activated transcription from Hor2 and Itr1 promoters through binding to the intact DOF motifs. When the two DOF factors are co-bombarded together an additive effect was observed upon the expression of the Itr1 gene. In-frame fusion of the Sad ORF to the reporter green fluorescent protein gene directs the fluorescence expression to the nucleus in transiently transformed onion epidermal layers. The visualization of fluorescence in the nucleus of onion cells, using the bimolecular fluorescent complex (BiFC) approach, has shown the in vivo interaction between SAD and the R2R3MYB protein GAMYB. The interaction in plant cells has also been documented for the DOF protein BPBF and GAMYB, but nuclear interaction could not be detected between BPBF and SAD by this procedure.

Cell Nucleus↗

Arterial pressure lability and renal sympathetic nerve activity are dissociated in SAD rats.

The purpose of this study was to determine whether the sympathetic nervous system drives the high variability of arterial pressure (AP) observed after sinoaortic denervation (SAD) in rats. One or fourteen days after SAD, rats were instrumented chronically to record mean AP (MAP), heart rate (HR), and renal sympathetic nerve activity (RSNA) in the conscious unrestrained state. Acute SAD increased MAP, HR, RSNA, and variability of MAP and decreased variability of both HR and RSNA. In rats with chronic SAD, variability of MAP remained high, whereas MAP, HR, RSNA, and variability of HR and RSNA returned to normal levels. Correlation analysis showed that, in sham-operated rats, AP and RSNA were negatively correlated in 90% of cases. In contrast, rats with both acute and chronic SAD exhibited only 30% negative and 25% positive correlations. These results indicate that 1) low AP variability in intact rats results from baroreflex-mediated inversely related fluctuations in RSNA and HR and 2) high variability of AP after acute and chronic SAD is correlated infrequently with RSNA. Because lability is reduced by interventions that block the sympathetic nervous system, we conclude that lability of AP associated with SAD appears to be mediated largely by a permissive role of sympathetic activity.

Animals↗

Treatment with oral clotrimazole blocks Ca(2+)-activated K+ transport and reverses erythrocyte dehydration in transgenic SAD mice. A model for therapy of sickle cell disease.

Prevention of red cell K+ and water loss is a therapeutic strategy for sickle cell disease. We have investigated in vitro and in vivo the effects of clotrimazole (CLT) and miconazole (MIC) on transgenic mice red cells expressing hemoglobin SAD. CLT blocked the Gardos channel (ID50 75 +/- 22 nM; n = 3) and the A23187-induced dehydration of Hbbs/Hbbthal SAD 1 mouse erythrocytes in vitro. Oral treatment with CLT (160 mg/kg per d) and MIC (100 mg/kg per d) inhibited the Gardos channel in both SAD 1 and control (Hbbs/Hbbthal) mice. In the SAD 1 mice only, cell K+ content increased, and mean corpuscular hemoglobin concentration and cell density decreased. After 7 d of treatment, the hematocrit of SAD 1, CLT-treated animals also increased. All changes were fully reversible. Long-term treatments of SAD 1 mice with oral CLT (80 mg/kg per d for 28 d) lead to sustained increases in cell K+ content and hematocrit and sustained decreases in mean corpuscular hemoglobin concentration and cell density, with no changes in animals treated with vehicle alone. Thus, CLT and MIC can reverse dehydration and K+ loss of SAD 1 mouse erythrocytes in vitro and in vivo, further supporting the potential utility of these drugs in the treatment of sickle cell anemia.

Administration, Oral↗

Brain activity during transient sadness and happiness in healthy women.

OBJECTIVE: The specific brain regions involved in the normal emotional states of transient sadness or happiness are poorly understood. The authors therefore sought to determine if H2(15)O positron emission tomography (PET) might demonstrate changes in regional cerebral blood flow (rCBF) associated with transient sadness or happiness in healthy adult women. METHOD: Eleven healthy and never mentally ill adult women were scanned, by using PET and H2(15)O, during happy, sad, and neutral states induced by recalling affect-appropriate life events and looking at happy, sad, or neutral human faces. RESULTS: Compared to the neutral condition, transient sadness significantly activated bilateral limbic and paralimbic structures (cingulate, medial prefrontal, and mesial temporal cortex), as well as brainstem, thalamus, and caudate/putamen. In contrast, transient happiness had no areas of significantly increased activity but was associated with significant and widespread reductions in cortical rCBF, especially in the right prefrontal and bilateral temporal-parietal regions. CONCLUSIONS: Transient sadness and happiness in healthy volunteer women are accompanied by significant changes in regional brain activity in the limbic system, as well as other brain regions. Transient sadness and happiness affect different brain regions in divergent directions and are not merely opposite activity in identical brain regions. These findings have implications for understanding the neural substrates of both normal and pathological emotion.

Adult↗

Chocolate eating in healthy men during experimentally induced sadness and joy.

The study compared influences of qualitatively different emotions on eating. Motivation to eat, affective responses to chocolate and chewing of chocolate were investigated in healthy normal weight males during experimentally induced emotions. Subjects abstained from eating 2 h (n = 24) or 8 h (n = 24) before testing. They received pieces of chocolate after viewing film clips presented to induce anger, fear, sadness and joy. Motivation to eat and most affective responses to eating chocolate were higher after 8 h than after 2 h of deprivation. Sadness and joy affected motivation to eat in opposite directions: joy increased and sadness decreased appetite (p < 0.001). In joy, a higher tendency to eat more chocolate was reported (p < 0.001), and chocolate tasted more pleasant (p < 0.001) and was experienced as more "stimulating" than in sadness (p < 0.01). No effects of deprivation could be found for chewing time and number of chews. Results indicate that the quality of emotions can affect motivation to eat and affective responses to eating chocolate. Our findings on decreased eating responses to sadness in healthy males and the contradictory increased eating responses to sadness reported by others supports two types of emotion-induced changes of eating: emotion-congruent modulation of eating and eating to regulate emotions.

Adult↗

Age-related differences in the ability to perceive sad facial expressions.

BACKGROUND AND AIMS: Research has shown that the ability to label negative emotions displayed by facial expressions declines with age. Such studies, however, have tended to adopt the Ekman and Friesen (1976) Caucasian faces as their emotional stimuli. The purpose of the current study was to examine whether the age differences in identifying negative emotions are also found using the more recent Japanese and Caucasian Facial Expressions of Emotion (JACFEE). METHODS: In Experiment 1, 29 younger and 29 older individuals performed a verbal labeling emotion identification task (happy, sad, angry, frightened, disgusted, surprised, contemptuous). The ability to identify each emotion as a function of ethnicity across the age groups was examined. In order to reduce the verbal decision-making load on the task, a second experiment was conducted in which 60 younger and 60 older participants performed an emotion-matching task (sad, angry, contemptuous). RESULTS: In Experiment 1, older adults showed a significant decrement in the ability to recognize sad faces compared with younger adults, but no age x face ethnicity interaction was found. In Experiment 2, age differences were found when making same/different judgments regarding two sad faces or a sad and a contemptuous face. CONCLUSIONS: Results suggest that aging has an impact upon perceiving sad facial expressions, that this effect is not mediated by own-race versus other-race faces, and that age effects are not attributable to differences in verbal decision-making.

Adult↗

Standardized mood induction with happy and sad facial expressions.

The feasibility of applying ecologically valid and socially relevant emotional stimuli in a standardized fashion to obtain reliable mood changes in healthy subjects was examined. The stimuli consisted of happy and sad facial expressions varying in intensity. Two mood-induction procedures (happy and sad, each consisting of 40 slides) were administered to 24 young healthy subjects, who were instructed to look at each slide (self-paced) and try to feel the happy or sad mood expressed by the person in the picture. On an emotional self-rating scale, subjects rated themselves as relatively happier during the happy mood-induction condition and as relatively sadder during the sad mood-induction condition. Conversely, they reported that they were less happy during the sad mood-induction condition and less sad during the happy mood-induction condition. The effects were generalized to positive and negative affect as measured by the Positive and Negative Affect Scale. The intraindividual variability in the effect was very small. In a retest study after 1 month, the mood-induction effects showed good stability over time. The results encourage the use of this mood-induction procedure as a neurobehavioral probe in physiologic neuroimaging studies for investigating the neural substrates of emotional experience.

Adult↗

Differential limbic--cortical correlates of sadness and anxiety in healthy subjects: implications for affective disorders.

BACKGROUND: Affective disorders are associated with comorbidity of depression and anxiety symptoms. Positron emission tomography resting-state studies in affective disorders have generally failed to isolate specific symptom effects. Emotion provocation studies in healthy volunteers have produced variable results, due to differences in experimental paradigm and instructions. METHODS: To better delineate the neural correlates of sad mood and anxiety, this study used autobiographical memory scripts in eight healthy women to generate sadness, anxiety, or a neutral relaxed state in a within-subject design. RESULTS: Sadness and anxiety, when contrasted to a neutral emotional state, engaged a set of distinct paralimbic-cortical regions, with a limited number of common effects. Sadness was accompanied by specific activations of the subgenual cingulate area (BA) 25 and dorsal insula, specific deactivation of the right prefrontal cortex BA 9, and more prominent deactivation of the posterior parietal cortex BAs 40/7. Anxiety was associated with specific activations of the ventral insula, the orbitofrontal and anterior temporal cortices, specific deactivation of parahippocampal gyri, and more prominent deactivation of the inferior temporal cortex BAs 20/37. CONCLUSIONS: These findings are interpreted within a model in which sadness and anxiety are represented by segregated corticolimbic pathways, where a major role is played by selective dorsal cortical deactivations during sadness, and ventral cortical deactivations in anxiety.

Adult↗

Dawn simulation and bright light in the treatment of SAD: a controlled study.

BACKGROUND: Some small controlled studies have found that dawn simulation is effective in treating seasonal affective disorder (SAD). With a larger sample size and a longer duration of treatment, we compared dawn simulation with bright light therapy and a placebo condition in patients with SAD. METHOD: Medication-free patients with SAD were randomly assigned to one of three conditions: bright light therapy (10,000 lux for 30 min, from 6:00 AM to 6:30 AM), dawn simulation (1.5 hour dawn signal from 4:30 AM to 6:00 AM peaking at 250 lux), and a placebo condition, a dim red light (1.5 hour dawn signal from 4:30 am to 6:00 AM peaking at 0.5 lux.) Over the subsequent 6 weeks, the subjects were blindly rated by a psychiatrist using the Structured Interview Guide for the Hamilton Depression Rating-Seasonal Affective Disorder Version (SIGH-SAD). We modeled the profiles of the remissions (SIGH-SAD < or = 8) and response (> or =50% decrease in SIGH-SAD) to treatment over time using Cox proportional hazards models. RESULTS: The sample consisted of 95 subjects who were randomized to the three conditions: bright light (n = 33), dawn simulation (n = 31) and placebo (n = 31). Dawn simulation was associated with greater remission (p <.05) and response (p <.001) rates compared to the placebo. Bright light did not differ significantly from the placebo. Dawn simulation was associated with greater remission (p <.01) and response (p <.001) rates compared to the bright light therapy. The mean daily hours of sunshine during the week before each visit were associated with a significant increase in likelihood of both remission (p <.001) and response (p <.001). CONCLUSIONS: Dawn simulation was associated with greater remission and response rates compared to the placebo and compared to bright light therapy. The hours of sunshine during the week before each assessment were associated with a positive clinical response.

Adult↗

Gender differences in neural correlates of recognition of happy and sad faces in humans assessed by functional magnetic resonance imaging.

To examine the effect of gender on the volume and pattern of brain activation during the viewing of alternating sets of faces depicting happy or sad expressions, 24 volunteers, 12 men and 12 women, participated in this functional magnetic resonance imaging study. The experimental stimuli were 12 photographs of Japanese adults selected from Matsumoto and Ekman's Pictures of Facial Affect. Four of these pictures depicted happy facial emotions, four sad, and four neutral. Half of the photographs were of men and the other half were of women. Consistent with previous findings, distinct sets of neural correlates for processing happy and sad facial emotions were noted. Furthermore, it was observed that male and female subjects used a rather different set of neural correlates when processing faces showing either happy or sad expressions. This was more noticeable when they were processing faces portraying sad emotions than happy emotions. Our findings provide some preliminary support for the speculation that the two genders may be associated with different areas of brain activation during emotion recognition of happy or sad facial expressions. This suggests that the generalizability of findings in regard to neural correlates of facial emotion recognition should consider the gender of the subjects.

Adult↗

Real-time fMRI of temporolimbic regions detects amygdala activation during single-trial self-induced sadness.

Temporolimbic circuits play a crucial role in the regulation of human emotion. A highly sensitive single-shot multiecho functional magnetic resonance imaging (fMRI) technique with gradient compensation of local magnetic field inhomogeneities and real-time data analysis were used to measure increases in amygdala activation during single 60-s trials of self-induced sadness. Six healthy male and female subjects performed a validated mood induction paradigm with randomized presentation of sad or neutral faces in 10 trials per scan. Subjects reported the intensity of experienced sadness after each trial. Immediate feedback of amygdala activation was given to the subjects during the ongoing scan to reinforce mood induction. Correspondence between increased intensity of predominantly left sided amygdala activation and self-rating of sadness was found in 78% of 120 sad trials, in contrast to only 14% of neutral trials. Amygdala activation was reproducible during repeated scanning sessions and displayed the strongest correlation with self-rating among all regions. These results suggest that amygdala activation may be closely associated with self-induced sadness. This novel real-time fMRI technology is applicable to a wide range of neuroscience studies, particularly those of the limbic system, and to neuropsychiatric conditions, such as depression, in which pathology of the amygdala has been implicated.

Adult↗

The impact of individual differences on the neural circuitry underlying sadness.

Several functional neuroimaging studies have been carried out in healthy subjects to investigate the neural correlates of sadness. Importantly, there is little consistency among the results of these studies. Hypothesizing that individual differences may account for the discrepancies among these investigations, we conducted two functional magnetic resonance imaging (fMRI) studies to identify the neural circuitry underlying this basic emotion. In these two methodologically identical studies, two different groups (n = 10 for each study) of healthy female subjects were scanned while they were experiencing a transient state of sadness induced by viewing sad film excerpts. In the first of these studies, sadness was correlated with significant loci of activation in the anterior temporal pole and insula (P < 0.05, corrected). In the second study, however, sadness was correlated with significant activation in the orbitofrontal and medial prefrontal cortices (P < 0.05, corrected). In addition, individual statistical parametric maps revealed a marked degree of interindividual variability in both Study 1 and Study 2. These results strongly support the view that individual differences may be responsible for the inconsistencies found in the literature regarding the neural substrates of sadness and of other basic emotions. These findings also suggest that individual data should be reported in addition to group data, because they provide useful information about the variability present in the subjects investigated and, thus, about the typicality and generalizability of the results.

Adult↗

Quality of life as an outcome indicator in patients with seasonal affective disorder: results from the Can-SAD study.

BACKGROUND: Although a host of studies have now examined the relationship between quality of life (QoL) and non-seasonal depression, few have measured QoL in seasonal affective disorder (SAD). We report here on results from the Can-SAD trial, which assessed the impact of treatment with either antidepressant medication or light therapy upon QoL in patients diagnosed with SAD. METHOD: This Canadian double-blind, multicentre, randomized controlled trial included 96 patients who met strict diagnostic criteria for SAD. Eligible patients were randomized to 8 weeks of treatment with either: (1) 10000 lux light treatment and a placebo capsule or (2) 100 lux light treatment (placebo light) and 20 mg fluoxetine. QoL was measured with the Quality of Life Enjoyment and Satisfaction Questionnaire (Q-LES-Q) and the Medical Outcomes Study (MOS) Short-Form General Health Survey (SF-20) at baseline and 8 weeks. RESULTS: Both intervention groups showed significant improvement in QoL over time with no significant differences being detected by treatment condition. Q-LES-Q scores increased significantly in seven of eight domains, with the average scores rising from 48 x 0 (S.D.=10 x 7) at baseline to 69 x 1 (S.D.=15 x 6) at week 8. Treatment-related improvement in QoL was strongly associated with improvement in depression symptoms. DISCUSSION: Patients with SAD report markedly impaired QoL during the winter months. Treatment with light therapy or antidepressant medication is associated with equivalent marked improvement in perceived QoL. Studies of treatment interventions for SAD should routinely include broader indices of patient outcome, such as the assessment of psychosocial functioning or life quality.

Adult↗

Self-assessed health, sadness and happiness in relation to the total burden of symptoms from the lower urinary tract.

OBJECTIVES: To evaluate the effect of lower urinary tract symptoms (LUTS) on self-assessed health, sadness and happiness of men. SUBJECTS AND METHODS: The study included 504 men (aged 40-80 years) in the rural community of Surahammar, Sweden, who a year earlier had reported stress incontinence, urgency or postvoid dribbling in answer to a postal questionnaire, and 504 age-matched control men from the same community. The occurrence of 12 specific LUTS was rated using the Danish Prostatic Symptom Score. Health, sadness and happiness were measured by three questions from the Medical Outcomes Study Short-Form 36 health survey questionnaire. RESULTS: Completed questionnaires were returned by 74.2% of men (748/1008). A low score for health was reported by 34% of men with one to four LUTS, by 67% with five to eight, and by 75% with nine or more LUTS. The total LUTS burden correlated with lower scores for happiness and with higher scores for sadness. For each of the 12 specific LUTS, men with the symptom had lower scores for health and happiness, and higher scores for sadness, than men without the symptom. Comparing men with the symptom of 'other incontinence' to men with no 'other incontinence', the relative risk (95% confidence interval) of impaired health was 2.2 (1.8-2.8), while that of a high score for happiness was 0.5 (0.3-0.7) and that of greater sadness was 2.3 (1.7-3.3). Social status, marital status, education, smoking, physical activity and urinary tract infection all affected the impact of LUTS. CONCLUSIONS: The total burden of LUTS is related to self-assessed health, sadness and happiness.

Adult↗

Neuroanatomical correlates of happiness, sadness, and disgust.

OBJECTIVE: Happiness, sadness, and disgust are three emotions that differ in their valence (positive or negative) and associated action tendencies (approach or withdrawal). This study was designed to investigate the neuroanatomical correlates of these discrete emotions. METHOD: Twelve healthy female subjects were studied. Positron emission tomography and [15O]H2O were used to measure regional brain activity. There were 12 conditions per subject: happiness, sadness, and disgust and three control conditions, each induced by film and recall. Emotion and control tasks were alternated throughout. Condition order was pseudo-randomized and counterbalanced across subjects. Analyses focused on brain activity patterns for each emotion when combining film and recall data. RESULTS: Happiness, sadness, and disgust were each associated with increases in activity in the thalamus and medial prefrontal cortex (Brodmann's area 9). These three emotions were also associated with activation of anterior and posterior temporal structures, primarily when induced by film. Recalled sadness was associated with increased activation in the anterior insula. Happiness was distinguished from sadness by greater activity in the vicinity of ventral mesial frontal cortex. CONCLUSIONS: While this study should be considered preliminary, it identifies regions of the brain that participate in happiness, sadness, and disgust, regions that distinguish between positive and negative emotions, and regions that depend on both the elicitor and valence of emotion or their interaction.

Adult↗

Regional cerebral blood flow changes during visually induced subjective sadness in healthy elderly persons.

This study examined regional cerebral blood flow (rCBF) changes associated with visually induced sad affect in healthy elderly persons. Subjects viewed sadness-laden, happiness-laden, and emotionally neutral image sets while rCBF was recorded using [(15)O] water PET. The sad image set included human faces and scenery/objects ("scenes"). To control for secondary sensory processing, the neutral and happy comparison sets included exclusively either human faces or scenes. During the sad condition, the ventral prefrontal and temporal cortices were more active compared with happy and neutral scenes conditions and the thalamus was more active compared with happy and neutral faces conditions. Ventral prefrontal cortex and thalamus were associated with processing of sad visual stimuli, whether compared with neutral or happy stimuli. The specific findings associated with sad affect were contingent on the comparison stimuli content (scenes or human faces), not affect (i.e., comparison with neutral or happy conditions).

Affect↗

Formation of dense erythrocytes in SAD mice exposed to chronic hypoxia: evaluation of different therapeutic regimens and of a combination of oral clotrimazole and magnesium therapies.

We have examined the effect of hydroxyurea (HU), clotrimazole (CLT), magnesium oxide (Mg), and combined CLT+Mg therapies on the erythrocyte characteristics and their response to chronic hypoxia in a transgenic sickle mouse (SAD) model. SAD mice were treated for 21 days with 1 of the following regimens (administered by gavage): control (n = 6), HU (200 mg/d; n = 6), CLT (80 mg/kg/d, n = 5), Mg (1,000 mg/kg/d, n = 5), and CLT+Mg (80 and 1,000 mg/kg/d, respectively, n = 6). Nine normal mice were also treated as controls (n = 3), HU (n = 3), and CLT+Mg (n = 3). Treatment with HU induced a significant increase in mean corpuscular volume and cell K content and a decrease in density in SAD mice. Treatment with the CLT and Mg, either alone or in combination, also increased cell K and reduced density in SAD mice. After 21 days of treatment, the animals were exposed to hypoxia (48 hours at 8% O(2)) maintaining the same treatment. In the SAD mice, hypoxia induced significant cell dehydration. These hypoxia-induced changes were blunted in either HU- or Mg-treated SAD mice and were completely abolished by either CLT or CLT+Mg treatment, suggesting a major role for the Gardos channel in hypoxia-induced dehydration in vivo.

Administration, Oral↗