Distinct forebrain and cerebellar isozymes of type II Ca^(2+)/calmodulin-dependent protein kinase associate differently with the postsynaptic density fraction

Forebrain and cerebellar Type II Ca2+/calmodulin-dependent protein kinases have different subunit compositions. The forebrain holoenzyme, characterized in our laboratory, is a 650-kDa holoenzyme composed of 50-kDa alpha-subunits and 60-kDa beta-subunits assembled in approximately a 3:1 ratio (Bennett, M. K., Erondu, N. E., and Kennedy, M. B. (1983) J. Biol. Chem. 258, 12735-12744). The cerebellar isozyme is a 500-kDa holoenzyme composed of alpha-subunits and beta-subunits assembled in almost the converse ratio, approximately four beta-subunits for each alpha-subunit. When compared by tryptic peptide mapping and by immunochemical techniques, the beta-subunits from the two brain regions are indistinguishable and the alpha-subunits appear closely related. The specific activities, substrate specificities, and catalytic constants of the cerebellar and forebrain isozymes are similar, suggesting that the alpha- and beta-subunits contain similar catalytic sites. However, two differences in the properties of the isozymes may result in functional differences between them in vivo. First, the apparent affinity of the cerebellar kinase for Ca2+/calmodulin is 2-fold higher than that of the forebrain kinase. Second, the two isozymes appear to associate differently with subcellular structures. Approximately 85% of the cerebellar kinase and 50% of the forebrain kinase remain in the particulate fraction after homogenization under standard conditions. However, they are present in different amounts in postsynaptic density fractions. Postsynaptic densities prepared from forebrain contain the forebrain isozyme. Immunochemical measurements show that it comprises approximately 16% of their total protein. In contrast, postsynaptic densities prepared from cerebellum contain the cerebellar isozyme, but it comprises only approximately 1-2% of their total protein. Thus, the alpha-subunit may play a role in anchoring Type II Ca2+/calmodulin-dependent protein kinase to postsynaptic densities

Activation of type II calcium/calmodulin-dependent protein kinase by Ca^(2+)/calmodulin is inhibited by autophosphorylation of threonine within the calmodulin-binding domain

It is now well established that autophosphorylation of a threonine residue located next to each calmodulin-binding domain in the subunits of type II Ca^(2+)/calmodulin-dependent protein kinase causes the kinase to remain active, although at a reduced rate, after Ca^(2+) is removed from the reaction. This autophosphorylated form of the kinase is still sensitive to Ca2+/calmodulin, which is required for a maximum catalytic rate. After removal of Ca^(2+), new sites are autophosphorylated by the partially active kinase. Autophosphorylation of these sites abolishes sensitivity of the kinase to Ca^(2+)/calmodulin (Hashimoto, Y., Schworer, C. M., Colbran, R. J., and Soderling, T. R. (1987) J. Biol. Chem. 262, 8051-8055). We have identified two pairs of homologous residues, Thr^(305) and Ser^(314) in the alpha subunit and Thr^(306) and Ser^(315) in the beta subunit, that are autophosphorylated only after removal of Ca^(2+) from an autophosphorylation reaction. The sites were identified by direct sequencing of labeled tryptic phosphopeptides isolated by reverse-phase high pressure liquid chromatography. Thr^(305-306) is rapidly dephosphorylated by purified protein phosphatases 1 and 2A, whereas Ser^(314-315) is resistant to dephosphorylation. We have shown by selective dephosphorylation that the presence of phosphate on Thr^(305-306) blocks sensitivity of the kinase to Ca^(2+)/calmodulin. In contrast, the presence of phosphate on Ser^(314-315) is associated with an increase in the Kact for Ca^(2+)/calmodulin of only about 2-fold, producing a relatively small decrease in sensitivity to Ca^(2+)/calmodulin

Condom Use among Young African American Men: Implications for Planning Interventions

Condom Use among Young African American Men: Implications for Planning Interventions Sexually transmitted diseases, including HIV, continue to present significant public health problems affecting young people in the United States, especially African Americans. While African Americans make up about 12% of the U.S. population, in 2010 they accounted for 44% of new HIV infections in 2010 and 48% of all persons living with AIDS in 2007. The 2010 data shows that of these new cases, 38% occurred among African American males ages 13-24 years old. Correct condom use remains a challenge in this population and efforts to increase condom use among minority males has been a formidable challenge. This paper reports the results of formative research conducted in order to guide the development of an intervention to increase consistent, effective condom use for young African American males. Methods: A snowball sampling approach was used to recruit participants. African American males, ages 18-24, who self-reported as sexually active were eligible to participate in one of four focus groups or one of six individual interviews. All study events were conducted at community locations. Each event was audiotaped and notes were taken. Analysis was performed using using NVivo-9. The coding strategy included emic and etic codes and a coding tree was developed which was used to identify themes. Results: A total of 36 African American males between the ages of 18-24 (mean 20.7 years) took part. In general, participants felt condom use was highly influenced by contextual factors including partner interest, partner communication, length of relationship and trust. Condom use was also influenced by a sense of invincibility and being caught up in the moment. Notably, most sexual activity occurred outside of a relationship, most often within the party scene or as quickly arranged hook-ups. Analysis: In order to ensure maximum impact on the development of the intervention, the results from this formative phase were viewed through the Transtheoretical Model of Behavior Change (TTM) and most participants would be described as being in the pre-contemplation or contemplation stages of behavioral change. While all participants expressed some understanding of the risks of unprotected sex, many did not connect risks to consequences. While the data did not indicate that condom use behavior was likely to change in the short-term (less than 6 months), several participants were contemplating making a change. Discussion: The snowball sampling approach allowed us to understand the participants’ social network and allowed us to consider social influences as well as about individual attitudes and beliefs. In the TTM frame, interventions designed for this population need to include contemplators and pre-contemplators and should focus on modification of cognition, affect and behaviors. Our research also shows that several of the underlying assumptions of TTM are at odds with the framework within which sex often occurs for this population and condom use decisions are highly influenced by the social context. In light of the results, the intervention placed condom use into a health promotion context. It combines group activities and one-on-one interaction. Group activities can impact shared values and beliefs and, thus, the intervention builds social support for behavior changes while addressing individual capacity

Diagnostic Radiology in Liberia: A Country Report

Liberia is a tropical country located south of the Sahara Desert in coastal West Africa. It lies at 6 °30’ North Latitude and 9° 30’ West Longitude and is bordered by Guinea, Cote d’Ivoire, Sierra Leone and the Atlantic Ocean. Liberia has three distinct topographical areas: 1) coastal plain, creeks, lagoons and mangrove swamps; 2) rolling, forested hills with elevations up to 500 feet that cover most of the country; and 3) low mountains and plateaus in the Northern highlands with elevations reaching 4,748 feet (Nimba Mountains). Liberia is home to approximately four million people and is roughly the size of the US state of Tennessee. Named after former US President James Monroe, Liberia’s capital Monrovia is a coastal city with a population of one million (1). There are two major seasons in Liberia: dry and rainy. The dry season occurs between December and March, and is is characterized by warm days and cool nights, with risk of sand storms from the Sahara Desert (2). The rainy season occurs between mid-April and mid-November. The average annual rainfall is 200 inches on the coast and decreases to 80 inches in areas farthest inland, and the average temperature is 27 degrees Celsius (81 degrees Fahrenheit) (1). Liberia is a low-income country that relies heavily on foreign aid (3). Liberia is the seventh poorest nation in the world, ranking 31st among 46 sub-Sarahan African countries in national income. In 2013, Liberia’s per capita GDP was $900 US (3). Liberia’s economy depends heavily on natural resources, with mining and agriculture being the dominant industries. Iron exportation has grown and in 2013 overcame rubber as Liberia’s top export. According to the 2013 Central Bank of Liberia (CBL) Annual Report, iron ore and rubber represent 82% of Liberia’s total exports (4). Civil war destroyed much of Liberia’s economy, including critical infrastructure in and around Monrovia. Although conditions are favorable for agriculture, Liberia does not produce nearly enough food to meet the demands of its population. The country imports large quantities of food, with rice alone accounting for 10% of its overall imports (5)

A New Limit on the Antiproton Lifetime

Measurements of the cosmic ray pbar/p ratio are compared to predictions from an inhomogeneous disk-diffusion model of pbar production and propagation within the Galaxy, combined with a calculation of the modulation of the interstellar cosmic ray spectra as the particles propagate through the heliosphere to the Earth. The predictions agree with the observed pbar/p spectrum. Adding a finite pbar lifetime to the model, we obtain the limit tau_pbar > 0.8 Myr (90 % C.L.).Comment: 13 pages, 3 encapsulated Postscript figures, uses AASTeX; accepted by Astrophysical Journal; minor change

Towards understanding the clinical significance of QT peak prolongation: a novel marker of myocardial ischemia independently demonstrated in two prospective studies

Background: QT peak prolongation identified patients at risk of death or non-fatal MI. We tested the hypothesis that QT peak prolongation might be associated with significant myocardial ischaemia in two separate cohorts to see how widely applicable the concept was. Methods and Results: In the first study, 134 stroke survivors were prospectively recruited and had 12-lead ECGs and Nuclear myocardial perfusion scanning. QT peak was measured in lead I of a 12-lead ECG and heart rate corrected by Bazett’s formula (QTpc). QTpc prolongation to 360ms or more was 92% specific at diagnosing severe myocardial ischaemia. This hypothesis-generating study led us to perform a second prospective study in a different cohort of patients who were referred for dobutamine stress echocardiography. 13 of 102 patients had significant myocardial ischaemia. Significant myocardial ischaemia was associated with QT peak prolongation at rest (mean 354ms, 95% CI 341-367ms, compared with mean 332ms, 95% CI 327-337ms in those without significant ischaemia; p=0.002). QT peak prolongation to 360ms or more was 88% specific at diagnosing significant myocardial ischaemia in the stress echocardiography study. QT peak prolongation to 360ms or more was associated with over 4-fold increase odds ratio of significant myocardial ischaemia. The Mantel- Haenszel Common Odds Ratio Estimate=4.4, 95% CI=1.2-16.0, p=0.023. Conclusion: QT peak (QTpc) prolongation to 360ms or more should make us suspect the presence of significant myocardial ischaemia. Such patients merit further investigations for potentially treatable ischaemic heart disease to reduce their risk of subsequent death or non-fatal MI

Transactional Sex among Youths in Post-conflict Liberia

This paper presents findings on sexual risk behaviours of Liberian youths based on five focus-group discussions conducted with 6th and 7th graders (n=36) attending an elementary/middle school in Monrovia, Liberia. The purpose of the focus-group discussions was to gain an understanding of the sexual behaviours of in-school Liberian adolescents. The focus-group discussions were part of a larger study to adapt an evi-dence-based HIV-prevention intervention—Making Proud Choices!—for in-school youths. Post-conflict conditions were discussed as a contributor to the emergence of high-risk sexual behaviours, including transactional sex, sexual violence, and lack of condom-use. Transactional sex was often described by the focus-group participants as occurring between young females and older, more financially-secure males to obtain cash, food, clothing, western commodities, and school-fees and was often encouraged by parents and promoted by peers. The findings also indicate that female adolescents make choices to engage in transactional sex to gain access to a continuum of material and consumer needs. These findings suggest that individual risk-taking behaviours are nested within complex sexual economies and that HIV-prevention interventions should be considered that leverage females’ agency and control