Contribution of Protein Intake and Concurrent Exercise to Skeletal Muscle Quality with Aging.

BACKGROUND: The use of magnetic resonance imaging (MRI) derived functional cross-sectional area (FCSA) and intramuscular adipose tissue (IMAT) to define skeletal muscle quality is of fundamental importance in order to understand aging and inactivity-related loss of muscle mass. OBJECTIVES: This study examined factors associated with lower-extremity skeletal muscle quality in healthy, younger, and middle-aged adults. DESIGN: Cross-sectional study. SETTING AND PARTICIPANTS: Ninety-eight participants (53% female) were classified as younger (20-35 years, n=50) or middle-aged (50-65 years, n=48) as well as sedentary (≤1 day per week) or active (≥3 days per week) on self-reported concurrent exercise (aerobic and resistance). MEASUREMENTS: All participants wore an accelerometer for seven days, recorded a three-day food diary, and participated in magnetic resonance imaging (MRI) of the lower limbs. Muscle cross-sectional area (CSA) was determined by tracing the knee extensors (KE) and plantar flexors, while muscle quality was established through the determination of FCSA and IMAT via color thresholding. RESULTS: One-way analysis of variance and stepwise regression models were performed to predict FCSA and IMAT. KE-IMAT (cm2) was significantly higher among sedentary (3.74 ± 1.93) vs. active (1.85 ± 0.56) and middle-aged (3.14 ± 2.05) vs. younger (2.74 ± 1.25) (p < 0.05). Protein intake (g•kg•day-1) was significantly higher in active (1.63 ± 0.55) vs. sedentary (1.19 ± 0.40) (p < 0.05). Sex, age, concurrent exercise training status, and protein intake were significant predictors of KE FCSA (R2 = 0.71, p < 0.01), while concurrent exercise training status and light physical activity predicted 33% of the variance in KE IMAT (p < 0.01). CONCLUSION: Concurrent exercise training, dietary protein intake, and light physical activity are significant determinants of skeletal muscle health and require further investigation to mitigate aging and inactivity-related loss of muscle quality.

Genome-wide linkage scan in a large bipolar disorder sample from the National Institute of Mental Health genetics initiative suggests putative loci for bipolar disorder, psychosis, suicide, and panic disorder.

We conducted a 9-cM genome scan in a large bipolar pedigree sample from the National Institute of Mental Health genetics initiative (1060 individuals from 154 multiplex families). We performed parametric and nonparametric analyses using both standard diagnostic models and comorbid conditions thought to identify phenotypic subtypes: psychosis, suicidal behavior, and panic disorder. Our strongest linkage signals (genome-wide significance) were observed on chromosomes 10q25, 10p12, 16q24, 16p13, and 16p12 using standard diagnostic models, and on 6q25 (suicidal behavior), 7q21 (panic disorder) and 16p12 (psychosis) using phenotypic subtypes. Several other regions were suggestive of linkage, including 1p13 (psychosis), 1p21 (psychosis), 1q44, 2q24 (suicidal behavior), 2p25 (psychosis), 4p16 (psychosis, suicidal behavior), 5p15, 6p25 (psychosis), 8p22 (psychosis), 8q24, 10q21, 10q25 (suicidal behavior), 10p11 (psychosis), 13q32 and 19p13 (psychosis). Over half the implicated regions were identified using phenotypic subtypes. Several regions - 1p, 1q, 6q, 8p, 13q and 16p - have been previously reported to be linked to bipolar disorder. Our results suggest that dissection of the disease phenotype can enrich the harvest of linkage signals and expedite the search for susceptibility genes. This is the first large-scale linkage scan of bipolar disorder to analyze simultaneously bipolar disorder, psychosis, suicidal behavior, and panic disorder.

Substance Dependence Severity Scale: reliability and validity for ICD-10 substance use disorders.

The Substance Dependence Severity Scale (SDSS) is a semistructured interview that assesses the severity of the DSM-IV diagnoses of dependence and abuse and the ICD-10 diagnoses of substance dependence and harmful use across a wide range of substances. Previous research has demonstrated that the SDSS' DSM-IV dependence scales are reliable and valid indicators of diagnostic severity. However, the ICD-10 scales have not been psychometrically tested. This study investigated the test-retest reliability, internal consistency, diagnostic concordance, and concurrent validity of the SDSS' ICD-10 dependence and harmful use scales in 180 (112 male and 68 female) treated substance users. Test-retest reliabilities for the ICD-10 dependence scales ranged from good to excellent for alcohol, cocaine, heroin, and cannabis. Test-retest reliabilities for the SDSS' ICD-10 harmful use scales were in the good range for alcohol, cocaine, and heroin and the poor to fair range for cannabis. Internal consistency, diagnostic concordance, and concurrent validity results were comparable to the test-retest findings. These results support the use of the SDSS for assessing the severity of the ICD-10 dependence and harmful use diagnoses.

Substance Dependence Severity Scale (SDSS): reliability and validity of a clinician-administered interview for DSM-IV substance use disorders.

No existing diagnostic interview assesses severity of dependence based on DSM-IV criteria across a range of substances. The Substance Dependence Severity Scale (SDSS) was designed to serve this purpose, consisting of substance-specific scales of both severity and frequency of DSM-IV criteria. This study investigated the reliability and validity of the SDSS. The test-retest reliability of the SDSS in 175 (112 male and 63 female) treated substance users ranged from good to excellent for alcohol, cocaine, heroin and sedatives (interclass correlation coefficients (ICCs)=0.75-0.88 for severity, 0.67-0.85 for frequency). Results for cannabis were lower, ranging from fair to good (ICCs=0.50-0.62). Results for joint rating and internal consistency reliability were comparable to test-retest findings. In addition to indicators of concurrent validity, scale applications are presented and discussed.

Concurrent and predictive validity of the Substance Dependence Severity Scale (SDSS).

This study investigated the concurrent and predictive validity of the Substance Dependence Severity Scale (SDSS), a clinician-administered interview designed to assess the severity and frequency of DSM-IV dependence symptoms for a range of substances. A total of 172 (107 males and 66 females) treated substance users participated in the study. Of those, 89% (n=153) received at least one follow-up interview within 1-6 months of an initial assessment. For alcohol, cocaine and heroin, convergent and discriminant validity was supported by significant relationships between SDSS scores at baseline and other baseline measures of substance use consequences, such as the Addiction Severity Index (ASI), as well as significant relationships between SDSS change scores from baseline to follow-up and change scores of other measures of consequences. SDSS scores were significantly associated with time to first post treatment use of alcohol, cocaine and heroin, although the nature of the associations was complex. Scale applications and areas for further study are discussed.

Motility potential of macaque epididymal sperm: the role of protein phosphatase and glycogen synthase kinase-3 activities.

Human and monkey ejaculated sperm contain protein phosphatase-1 (PP1), PP1 inhibitor 2 (12), and glycogen synthase kinase-3 (GSK-3). Inhibition of ejaculated human sperm protein phosphatase (PP) activity with calyculin-a (CL-A) significantly stimulates motility, implicating protein dephosphorylation in motility regulation. The present experiments were conducted to characterize and compare PP and GSK-3 activity in monkey caput and caudal epididymal sperm, to determine the cellular distribution of these enzymes, and to test the thesis that epididymal sperm PP activity is inversely related to motility. Caput epididymal sperm populations, (8.8% motile) contained levels of PP activity that were >3 times as high as those of caudal spermatozoa. This PP activity was further identified by inhibitor response profiles as PP1. In both caput and caudal sperm, the majority of this PP1 activity was localized in 100,000 x g soluble fractions. Western blot analysis indicated that a portion of this difference was the result of elevated amounts of PP1 in caput compared with caudal epididymal sperm. The presence of GSK-3 activity was undetectable in 100,000 x g insoluble fractions of epididymal sperm, whereas both caput and caudal sperm soluble fractions contained GSK-3 activity, which was approximately threefold higher in caput sperm compared with caudal populations. Treatment of caput epididymal sperm from the rhesus macaque with the PP inhibitor CL-A resulted in a significant, dose-dependent increase from 8 to 38% motile cells (without any effect on their path velocity). In contrast, CL-A had no significant influence on either percent motility or path velocity of caudal epididymal sperm. Cytosolic PP1 and GSK-3 activities appear to be inversely related to the motility of monkey epididymal sperm and may have a regulatory role in the development of the potential for motility in epididymal sperm.

Psychiatric research interview for substance and mental disorders: phenomenologically based diagnosis in patients who abuse alcohol or drugs.

The Psychiatric Research Interview for Substance and Mental Disorders (PRISM) is a psychiatric diagnostic interview designed to diagnose DSM-IV substance and mental disorders in patients who abuse alcohol or drugs. Primary disorders tested in the DSM-III-R version of the interview showed improved reliability over existing instruments, and substantially improved reliability for major depressive disorder (MDD). Developments for DSM-IV include a systematic set of procedures for differentiating primary disorders, substance-induced disorders, and the expected effects of intoxication and withdrawal based on the phenomenology of symptoms in conjunction with alcohol and drug use. A longitudinal version of the PRISM provides data on remission and relapse that can be analyzed with survival methods. Pilot and preliminary testing of the DSM-IV and longitudinal versions of the instruments is presented. By making use of psychometric principles, particularly the need to reduce criterion variance, these instruments can clarify some of the longstanding issues in the diagnosis of patients who abuse alcohol and drugs.

A tyrosine-phosphorylated 55-kilodalton motility-associated bovine sperm protein is regulated by cyclic adenosine 3',5'-monophosphates and calcium.

Sperm motility is regulated by protein phosphorylation. We have recently shown that a serine/threonine phosphatase system is involved in motility regulation. Two of the components of the phosphatase system, GSK-3 and PP1gamma2, are regulated by tyrosine phosphorylation. During our investigation of sperm tyrosine-phosphorylated proteins we discovered a 55-kDa protein whose tyrosine phosphorylation correlates closely to the motility state of sperm. This protein is tyrosine phosphorylated to a much higher degree in motile caudal than in immotile caput epididymal sperm. Motility inhibition of caudal epididymal sperm by protein kinase A (PKA) anchoring inhibition or by ionomycin-induced calcium overload led to the virtual disappearance of tyrosine phosphorylation of the 55-kDa protein. Conversely, treatment of sperm with motility activators, isobutylmethylxanthine or 8-bromo-cAMP, resulted in increased tyrosine phosphorylation of the protein. The protein was present in the soluble 100 000 x g supernatants of sperm extracts and was heat labile. Chromatography through diethylaminoethyl-cellulose and Western blot analysis showed that this 55-kDa protein is not a regulatory subunit of PKA or alpha-tubulin. Our results represent the identification of a soluble protein whose tyrosine phosphorylation varies directly with motility and suggest that motility regulation may involve cross talk between PKA, calcium, and tyrosine kinase pathways.

Psychiatric Research Interview for Substance and Mental Disorders (PRISM): reliability for substance abusers.

OBJECTIVE: The purpose of this study was to investigate the reliability of a new semistructured diagnostic interview, the Psychiatric Research Interview for Substance and Mental Disorders (PRISM), for substance-abusing patients. The reliability of psychiatric diagnoses for individuals who drink heavily or use drugs has been shown to be problematic. The PRISM was designed to improve the reliability for such individuals. METHOD: A test-retest reliability study of the PRISM was conducted with 172 patients being treated in dual-diagnosis or substance abuse settings. RESULTS: Good to excellent reliability was shown for many diagnoses, including affective disorders, substance use disorders, eating disorders, some anxiety disorders, and psychotic symptoms. The interview has recently been updated for DSM-IV diagnoses. CONCLUSIONS: The PRISM offers a method of producing psychiatric diagnoses with improved reliability for patients and other research subjects who have problems with alcohol or drugs.

Sperm motility development in the epididymis is associated with decreased glycogen synthase kinase-3 and protein phosphatase 1 activity.

Immotile bovine caput epididymal sperm contain levels of protein phosphatase activity twofold higher than do mature motile caudal sperm. Comparison of the inhibition profiles of endogenous phosphatase activities detected by okadaic acid (OA) and calyculin A (CA) revealed a pattern consistent with the predominance of a type 1 protein phosphatase (PP1). Immunoblot analysis identified PP1 gamma 2 (the testis-specific isoform of PP1) as the only PP1 isoform in sperm and showed little protein phosphatase 2A (PP2A). In addition, of the known PP1 inhibitors, i.e., DARPP-32, inhibitor 1 (I1), and inhibitor 2 (I2), only I2-like activity was detected in sperm. Inhibition of PP1 by the heat-stable I2-like activity purified from sperm could be reversed with purified glycogen synthase kinase-3 (GSK-3). Furthermore, sperm extracts contain an inactive complex of PP1 and I2 (termed PP1I) that could also be activated by purified GSK-3. The presence of GSK-3 in sperm was demonstrated by activation of purified PP1I, and quantitation revealed that immotile caput sperm contained sixfold higher GSK-3 activity than motile caudal sperm. Immunoblot analysis confirmed the expression of GSK-3 in sperm and revealed the occurrence of both the alpha and beta isoforms. Our findings suggest that the higher PP1 activity measured in immotile sperm, presumably due to higher GSK-3 activity, is responsible for holding motility in check. This conclusion was supported by the observation that the phosphatase inhibitors OA and CA, at micromolar and nanomolar levels, respectively, were able to induce motility in completely immotile bovine caput epididymal sperm and to stimulate the kinetic activity of mature caudal sperm. The intrasperm levels of cAMP, pH, and calcium were unaltered by treatment with these inhibitors. The results suggest a biochemical basis for the development and regulation of sperm motility and a possible physiological role for the PP1/I2/GSK-3 system.

Primate sperm contain protein phosphatase 1, a biochemical mediator of motility.

Sperm motility initiation, capacitation, and hyperactivation are modulated by an interplay of intracellular Ca2+, cAMP, and pH. Mechanisms by which these mediators alter sperm function have not been elucidated but may involve reversible alterations in regulatory protein phosphorylation. Studies were designed 1) to investigate the influence of the protein phosphatase (PP) inhibitor calyculin A (CA) on human sperm motility and 2) to characterize the CA-sensitive PP and its endogenous regulators in human and rhesus monkey sperm. CA (50 nM) treatment of human sperm resulted in an increase in percentage motility and an acceleration in mean path velocity. Inhibition of either protein phosphatase-1 (PP1) or protein phosphatase-2A (PP2A) could be responsible for this motility stimulation, since both of these phosphatases are sensitive to nanomolar quantities of CA. PP activity in human (n = 4) and rhesus monkey (n = 4) sperm sonicates was measured using [32P]-phosphorylase-a, the preferred substrate for PP1 and PP2A, in the absence of divalent cations. Human (6.2 +/- 4.5 x 10(-2) mU/10(6) sperm) and monkey (4.3 +/- 0.8 x 10(-2) mU/10(6) sperm) sonicates contained activity tentatively identified as PP1 on the basis of inhibition profiles in okadaic acid (OA) and CA. Western blot analysis with antibodies against various isoforms of PP1 subsequently documented the presence of PP1 gamma 2 in human and monkey sperm. PP1 activity in most tissues is regulated by the heat-stable inhibitors I1 or I2. Sperm sonicates contained inhibitor activity similar to I2 as well as glycogen synthase kinase-3 (GSK-3) activity, which is involved in the activation of the PP1-I2 complex. These results indicate, for the first time, that human and rhesus monkey sperm contain PP1 and regulators of PP1 and that inhibition of PP1 activity by CA can enhance motility.

Evidence against a functional ATP-dependent calcium extrusion mechanism in bovine epididymal sperm.

Bovine epididymal sperm resuspended in ionic buffers take up relatively large amounts of calcium. This uptake, which is almost entirely mitochondrial, apparently bypasses the sperm cytosol. The direct mitochondrial loading is an unusual aspect of sperm calcium uptake, which suggests that the plasma membrane region surrounding the mitochondria should be highly permeable to calcium, whereas the membrane domains surrounding the head and tail regions of sperm should be impermeable. This study was undertaken to determine the role of a plasma membrane calcium ATPase in sperm calcium homeostasis. Kinetics of calcium (45Ca2+) uptake into intact and permeabilized caudal epididymal sperm confirmed that mitochondrial calcium uptake occurs with virtually no resistance from the surrounding plasma membrane. Cytoplasmic calcium accumulation by sperm depleted of intracellular ATP, measured in the presence of mitochondrial calcium uptake inhibitors, showed no increase upon energy depletion as would be expected if an ATP-dependent calcium extrusion mechanism were present. Furthermore, lowering the incubation temperature to further reduce the activity of the calcium ATPase in these energy-depleted sperm was also without effect on calcium accumulation. The calcium ATPase inhibitor vanadate, even at high concentrations, failed to increase intracellular 45Ca2+ accumulation. However, vanadate was effective in inhibiting motility showing that the compound was accumulated into sperm to inhibit flagellar dyenin ATPase. Therefore, the lack of effect of vanadate on 45Ca2+ accumulation was not due to its inability to enter sperm. Other calcium ATPase inhibitors such as quercetin, thapsigargin, and cyclopiazonic acid, which readily demonstrate ATP-dependent calcium extrusion in other somatic cells, were also without effect on sperm calcium accumulation.(ABSTRACT TRUNCATED AT 250 WORDS)

Quantification of area at risk during coronary occlusion and degree of myocardial salvage after reperfusion with technetium-99m methoxyisobutyl isonitrile.

Serial myocardial imaging with technetium-99m methoxyisobutyl isonitrile (99mTc-MIBI) has been proposed for evaluating myocardial salvage after reperfusion. To define 99mTc-MIBI uptake before and after reperfusion, 17 open-chest dogs underwent 3 hours of left anterior descending artery occlusion and 3 hours of reperfusion. 99mTc-MIBI was injected during occlusion (group 1) or after 90 minutes of reperfusion (group 2). Myocardial 99mTc-MIBI activity was correlated with microsphere flow during occlusion and reperfusion. Anatomic risk area and infarct area were defined by postmortem vital staining and correlated with the perfusion defects defined by analysis of 99mTc-MIBI macroautoradiographs and gamma camera images of myocardial slices. The left ventricle was divided into 96 segments for gamma well counting. Flow and 99mTc-MIBI activity were normalized to nonischemic values. Myocardial segments were grouped, based on occlusion flow, into zones: severely ischemic (less than or equal to 30% nonischemic), moderately ischemic (greater than 30%, less than or equal to 60% nonischemic), mildly ischemic (greater than 60%, less than or equal to 90% nonischemic), and nonischemic (greater than 90%, less than or equal to 120% nonischemic). Among dogs injected with 99mTc-MIBI during coronary occlusion (group 1), myocardial 99mTc-MIBI activity correlated linearly with occlusion flow for both endocardial (r = 0.91) and transmural (r = 0.91) segments. The risk area defined by 99mTc-MIBI autoradiography (group 1) correlated with the postmortem risk area (rho = 0.94) but was 29% smaller than the anatomic risk area (p = 0.03), reflecting the contribution of collateral flow. Among dogs injected with 99mTc-MIBI after reperfusion (group 2), myocardial 99mTc-MIBI did not correlate with reperfusion flow in either endocardial or transmural segments. Among group 2 dogs, myocardial 99mTc-MIBI activity was significantly less than reperfusion flow at the time of injection in the severely ischemic (25 +/- 5% versus 74 +/- 24% nonischemic, p = 0.002), moderately ischemic (54 +/- 12% versus 96 +/- 15% nonischemic, p = 0.001), and mildly ischemic (84 +/- 6% versus 93 +/- 3% nonischemic, p = 0.002) zones. The defect area defined by 99mTc-MIBI autoradiography (group 2) correlated very closely with the postmortem infarct area (rho = 0.98). Thus, the myocardial uptake of 99mTc-MIBI during coronary occlusion correlates with occlusion flow and reflects the "area at risk." When 99mTc-MIBI was given after 90 minutes of reperfusion following 3 hours of coronary occlusion, the myocardial activity was significantly reduced compared with reperfusion flow in both necrotic and perinecrotic regions, reflecting myocardial viability more than the degree of reperfusion.