Amino acid sequences of myosin essential and regulatory light chains from two clam species: Comparison with other molluscan myosin light chains

Summary We have determined the amino acid sequences of the essential light chains (ELC) and regulatory light chains (RLC) of myosin from two species of clam,Mercenaria mercenaria andMacrocallista nimbosa, using protein chemistry methods. The N-termini of all four proteins were blocked, and sequencing was carried out on various chemically and enzymatically produced peptide fragments. Cleavage of eitherMercenaria RLC (MRLC) orMacrocallista RLC (VLC) at its 3 Arg yielded four peptides, three of which could not be sequenced directly, due to an N-terminal blocking group and 2 Arg-Gln bonds in these proteins. The fourth peptide was partially and specifically cleaved at an unusually reactive residue, Met-64, which is invariant in all known RLC sequences. A comparison of all available molluscan ELC and RLC sequences was carried out in search of clues to functionally important features of these proteins in muscles which are regulated by a Ca²⁺-sensitive myosin. By analogy with other RLCs, VRLC and MRLC may be phosphorylated at Ser-11 by an endogenous kinase. All myosin light chains, like troponin C and calmodulin, contain four homologous regions, I to IV, each of which contains a twelve-residue potential Ca²⁺-binding loop flanked on either side by a pair of helices. All RLCs, including those from Ca²⁺-insensitive myosins, contain a divalent cation-binding site in region I. Clam and other molluscan ELCs contain a single Ca²⁺-binding site in region III. This site is present only in the ELCs of myosins that are regulated by direct binding of Ca²⁺. The ELC site III is likely to play a key role in the regulation of molluscan muscle contraction.     

Pre- and postnatal depression and coping: a comparative approach

BACKGROUND: The assessment of perinatal depressions and coping style. Methods: With depression scales (EPDS, GHQ.12) and Carver's BriefCope, the authors compared the prevalence rates of pre and postnatal depression in a cohort of 277 French women. RESULTS: Their work revealed very high levels of prenatal depression (almost 20%) and less important but nonetheless sizeable rates (11%) of postnatal depression, making such perinatal depressions a major public health concern. The coping styles proposed in Carver's public health BriefCope questionnaire make it possible to significantly differentiate during these two periods between depressive women and their non-depressed counterparts. CONCLUSIONS: This enables us to underline factors of risk and protection suggesting the importance of setting up compensatory and preventive systems and evaluating their pertinence in the framework of future research.     

Dendritic cells generated in the presence of GM-CSF plus IL-15 prime potent CD8+ Tc1 responses in vivo

Dendritic cells (DC) comprise a system of professional antigen-presenting cells, which induce the stimulation of very rare antigen-specific naive T cells. DC progenitors can be stimulated to differentiate into immature DC by various growth factors, including GM-CSF and IL-4. Here we show that IL-15, in combination with GM-CSF, is a growth factor for murine DC. Murine bone marrow cells, depleted of T cells, B cells, I-A+ cells and Gr-1+ granulocytes, and cultured in the presence of GM-CSF plus IL-15 (IL-15 DC), yielded DC expressing high levels of CD11c and MHC class II molecules, as well as CD11b. These cells expressed significant levels of CD40, CD80 and CD86, and could stimulate allogeneic CD4+ T cells efficiently. Interestingly, IL-15 DC were far superior to DC generated with GM-CSF plus IL-4 in stimulating allogeneic CD8+ T cells in vitro. Consistent with this, IL-15 DC induced much more potent antigen-specific CD8+ T cell responses with high levels of Th1 cytokines in vivo, compared to DC generated with GM-CSF plus IL-4, or with GM-CSF plus TGF-beta, or with GM-CSF alone. Together, these data suggest that IL-15 promotes the development of DC, which induce potent Th1 and Tc1 responses in vivo. This suggests potential roles for these IL-15 DC cells in the immunotherapy of tumors and infectious diseases.     

Assessing the effects of bupropion SR on mood dimensions of depression

BACKGROUND: We assessed the therapeutic effects of bupropion SR and placebo on mood and anxiety symptoms derived from the tripartite model of mood. Based on evidence indicating linkages between dopaminergic activity and the emotional dimension of positive affect/anhedonia, we hypothesized that the dopaminergic effects of bupropion SR would yield particularly pronounced effects on symptoms of anhedonia, relative to anxiety. METHODS: Nineteen depressed outpatients were randomly assigned to treatment with either bupropion SR 300 mg/day or placebo during a 6-week initial treatment phase. This was followed by a second open-label phase in which patients previously treated with bupropion SR had their dose increased to 400 mg/day, and the placebo group was initiated on bupropion SR 300 mg/day. RESULTS: Random regression analyses revealed that during the initial double-blind phase, bupropion SR elicited greater declines than placebo on all measures except those that assessed anxiety. By contrast, the weakest placebo effects were evident on anhedonia. Items assessing the low positive affect pole of the anhedonia dimension were more sensitive to earlier/lower dose bupropion SR treatment, whereas items assessing the high positive affect pole were more sensitive to later/higher dose bupropion SR treatment. LIMITATIONS: Replication and extension using a larger sample size are mandated. CONCLUSIONS: This study suggests that the catecholaminergic effects of bupropion SR tended to produce more robust effects on anhedonia/positive affect than placebo.     

What and where in human audition: selective deficits following focal hemispheric lesions

A sound that we hear in a natural setting allows us to identify the sound source and localize it in space. The two aspects can be disrupted independently as shown in a study of 15 patients with focal right-hemispheric lesions. Four patients were normal in sound recognition but severely impaired in sound localization, whereas three other patients had difficulties in recognizing sounds but localized them well. The lesions involved the inferior parietal and frontal cortices, and the superior temporal gyrus in patients with selective sound localization deficit; and the temporal pole and anterior part of the fusiform, inferior and middle temporal gyri in patients with selective recognition deficit. These results suggest separate cortical processing pathways for auditory recognition and localization. Electronic Publication     

Substance abuse treatment effectiveness of publicly funded clients in Tennessee

The Tennessee Outcomes for Alcohol and Drug Services (TOADS) in collaboration with the Bureau of Alcohol and Drug Abuse Services at the Tennessee Department of Health evaluated the effectiveness of publicly funded substance abuse treatment programs in Tennessee by collecting and analyzing data from clients treated between 1998 and 2000. Using a structured questionnaire, TOADS staff conducted telephone interviews with clients 6 months after their admission to treatment facilities. The sample populations for these follow-up interviews ranged from 1,150 to 1,350 clients over the 3 years, and each year, post-treatment abstinence rates were around 60%, which suggests that treatment in Tennessee has been successful in reducing substance abuse. In addition, the follow-up interview data suggest that treatment also helped drastically reduce both unemployment and arrests among clients. These findings in Tennessee are comparable to treatment outcomes in other states. In addition to the positive effects that treatment has on clients, treatment is also cost-effective for state budgets since treatment reduces many of the burdens substance abuse places on the criminal justice system, the healthcare system, and other state-supported services.     

Are primed CD4+ T lymphocytes different from unprimed cells?

Primed and unprimed lymphocytes are usually classified as separate subsets of cells, based on phenotypic and functional distinctions. In the case of CD4⁺ T lymphocytes, primed cells are thought to proliferate more vigorously, quickly and easily, and to release a different profile of cytokines, than their naive equivalent. However, most of these data were obtained from studies in which populations of lymphocytes were compared before and after antigenic stimulation, and therefore did not distinguish between the effects resulting from the clonal expansion of specific precursor cells within such populations and those due to cell differentiation per se. We have investigated the contribution of precursor cell frequency to some of the functional changes observed in populations of CD4⁺ T cells following antigenic stimulation, using approaches in which antigen-specific precursor frequencies are high in both primary and secondary stimulations: mixed leukocyte reaction responses and cells from αβ T cell receptor transgenic mice. Our data suggest that when equivalent numbers of antigen-specific naive and previously primed CD4⁺ responder T cells are compared, there is no difference in their potency to proliferate but only the previously activated subset can generate cytokines such as interferon-γ.     

Controlling the spatial distribution of ECM components in degradable PEG hydrogels for tissue engineering cartilage

In developing a scaffold to support new tissue growth, the degradation rate and mass loss profiles of the scaffold are important design parameters. In this study, hydrogels were prepared by copolymerizing a degradable macromer, poly(lactic acid)-b-poly(ethylene glycol)-b-poly(lactic acid) endcapped with acrylate groups (PEG-LA-DA) with a nondegradable macromer, poly(ethylene glycol) dimethacrylate (PEGDM). The resulting hydrogels exhibited a range of degradation behavior and mass loss profiles. Chondrocytes were photoencapsulated in gels formulated with 50:50, 25:75, and 15:85 (mol % PEGDM: mol % PEG-LA-DA) and cultured for 6 weeks in vitro. The neocartilaginous tissue formed was examined biochemically and histologically. After 6 weeks, the DNA content in gels with 75 and 85% degradable crosslinks was nearly twice that of the DNA content in the 50% gels. The total collagen content was significantly higher in the 85% gel [2.4 +/- 0.8% wet weight (ww)] compared to the 50% gel (0.22 +/- 0.29% ww). In examining the neocartilaginous tissue with immunohistochemistry, type II collagen was localized in the pericellular region in the 50% gel; however, when increased degradation was incorporated into the gel, type II collagen was found throughout the neotissue. In summary, the important role of hydrogel degradation in controlling and influencing the deposition and distribution of extracellular matrix molecules was demonstrated and quantified.