The DNA double-strand break response in the nervous system

DNA double-strand breaks (DSBs) require a coordinated molecular response to ensure cellular or organism survival. Many factors required for the DSB response, including those involved in non-homologous end joining (NHEJ) and homologous recombination repair (HRR) are essential during nervous system development. Additionally, human syndromes resulting from defective responses to DNA damage often feature overt neuropathology such as neurodegeneration. Thus, appropriate responses to DSBs are critical for the normal development and maintenance of the nervous system.     

Human interleukin 24 (MDA-7/IL-24) protein kills breast cancer cells via the IL-20 receptor and is antagonized by IL-10

The melanoma differentiation-associated gene-7 (mda-7/IL-24) is a unique member of the interleukin 10 (IL-10) family of cytokines, with ubiquitous tumor cell pro-apoptotic activity. Recent data have shown that IL-24 is secreted as a glycosylated protein and functions as a pro-Th1 cytokine and as a potent anti-angiogenic molecule. In this study, we analyzed the activity of Ad-mda7 and its protein product, secreted IL-24, against human breast cancer cells. We show that Ad-mda7 transduction of human breast cancer cells results in G₂/M phase cell cycle arrest and apoptotic cell death, which correlates with secretion of IL-24 protein. Neutralizing antibody against IL-24 significantly inhibited Ad-mda7 cytotoxicity. IL-24 and IL-10 both engage their cognate receptors on breast cancer cells resulting in phosphorylation and activation of STAT3, however, IL-10 receptor binding failed to induce cell killing, indicating that tumor cell killing by IL-24 is independent of STAT3 phosphorylation. Treatment with exogenous IL-24 induced apoptosis in breast cancer cells and this effect was abolished by addition of anti-IL-24 antibody or anti-IL-20R1, indicating that bystander cell killing is mediated via IL-24 binding to the IL-20R1/IL-20R2 heterodimeric receptor complex. Co-administration of the related cytokine IL-10 inhibited killing mediated by IL-24 and concomitantly inhibited IL-24 mediated up-regulation of the tumor suppressor proteins, p53 and p27^Kip1. In summary, we have defined a tumor-selective cytotoxic bystander role for secreted IL-24 protein and identified a novel receptor-mediated death pathway in breast cancer cells, wherein the related cytokines IL-24 and IL-10 exhibit antagonistic activity.     

A nonneutralizing anti-HIV-1 antibody turns into a neutralizing antibody when expressed on the surface of HIV-1-susceptible cells: a new way to fight HIV

During HIV-1 infection or vaccination, HIV-1 envelope spikes elicit Ab responses. Neutralizing Abs block viral entry by recognizing epitopes on spikes critical for their interaction with receptor, coreceptors or fusion. In contrast, nonneutralizing Abs fail to do so because they recognize epitopes either buried or exposed but not critical for viral entry. Previously, we produced a high-affinity human mAb against the cluster II determinant of gp41. This Ab or its recombinant Fab and single-chain Fv have been repeatedly shown to bind to HIV-1 gp160 or gp41, but fail to block viral entry. We report that, surprisingly, expression of this nonneutralizing anti-HIV-1 gp41 single-chain Fv on the surface of human CD4 T cells markedly inhibits HIV-1 replication and cell-cell fusion. The inhibition targets the HIV-1 envelope at the level of viral entry, regardless of HIV-1 tropism. Although this bona fide nonneutralizing Ab does not neutralize HIV-1 entry when produced as a soluble protein, it acts as a neutralizing Ab when expressed on the cell surface. Expressing Abs on the surface of HIV-1-susceptible cells can be a new way to fight HIV-1.     

Human polyreactive and monoreactive antibodies: effect of glycosylation antigen binding

The present experiments were initiated to determine whetherthe carbohydrate portions of antibody molecules contribute topolyreactivity. Cell lines making human monoclonal polyreactiveor monoreactive antibodies of the immunoglobulin (Ig) M, IgGand IgA isotypes were treated with tunicamycin to block N-linkedglycosylation of the proteins. Analysis of the secreted nativeand non-glycosylated proteins revealed a >95% inhibitionof [³H]mannose incorporation. Electrophoresis on sodium dodecylsulphate-polyacrylamide gels of the proteins from tunicamycin-treatedcells showed increased mobility and the absence of [³H]mannoseincorporation of the immunoglobulin heavy chains, consistentwith the lack of glycosylation. The native and non-glycosylatedantibodies were then tested for their ability to bind differentantigens. Despite the lack of glycosylation, both polyreactiveand monoreactive antibodies bound to antigens with little ifany loss of reactivity or specificity. It is concluded thatthe carbohydrate moieties do not contribute significantly topolyreactivity. antigen binding glycosylation polyreactive antibodies     

Development of reproductive potential in protogynous coral reef fishes within Philippine no-take marine reserves

Empirical evidence for increases in the reproductive potential (egg output per unit area) of coral reef fish in no-take marine reserves (NTMRs) is sparse. Here, we inferred the development of reproductive potential in two species of protogynous reef fishes, Chlorurus bleekeri (Labridae: Scarinae) and Cephalopholis argus (Epinephelidae), inside and outside of Philippine NTMRs. We estimated key reproductive parameters and applied these to species-specific density and length data from 17 NTMRs (durations of protection 0–11 years) and paired fished sites (controls) in a space-for-time substitution approach. For C. argus, we also used density and length data collected almost annually over 29 years from a NTMR and an adjacent control at Apo Island. The results suggest that C. bleekeri can develop 6.0 times greater reproductive potential in NTMRs than controls after 11 years of protection, equivalent to approximately 582,000 more eggs produced 500 m⁻² inside NTMRs. Enhancement of reproductive potential in C. argus was not evident after 11 years in the space-for-time substitution. At Apo Island NTMR, reproductive potential of C. argus increased approximately 6-fold over 29 years but NTMR/control ratios in reproductive potential decreased through time (from 3.2 to 2.4), probably due to spillover of C. argus from the NTMR to the control. C. argus was estimated to produce approximately 113,000 more eggs 500 m⁻² inside Apo Island NTMR at the 29th year of protection. Ratios of reproductive potential between NTMR and controls in C. bleekeri and C. argus were often greater than corresponding ratios in density or biomass. The study underscores the importance of species-specific reproductive life history traits that drive variation in the development of larval fish subsidies that originate from NTMRs.     

Geographic variability in the gonadal development and sexual ontogeny of Hemigymnus,Cheilinus and Oxycheilinus wrasses among Indo-Pacific coral reefs

Patterns of reproductive ontogeny in four species of coral reef wrasses (F: Labridae) Hemigymnus melapterus, Hemigymnus fasciatus, Cheilinus fasciatus and Oxycheilinus digramma were investigated. Populations of each species were sampled from two island groups of the central Great Barrier Reef (GBR), Australia, and from coral reefs in the central Philippines. These three sampling locations span 30° of latitude. The GBR and Philippine reefs experience biologically significant differences in water temperature, geography and human activity. The studied wrasses are effectively unfished in Australia but heavily fished in the Philippines. Gonad weights, histology and demographic data were obtained across the entire size and age range of H. melapterus, C. fasciatus and O. digramma from all locations. Analysis identified three processes of male recruitment: functional gonochorism and both forms of protogynous hermaphroditism, monandry and diandry. The expression of these distinct sexual ontogenies was locality dependent. Populations of H. melapterus, H. fasciatus, C. fasciatus and O. digramma on the GBR showed consistently uniform patterns of sexual ontogeny, with all species being exclusively monandric. H. melapterus, C. fasciatus and O. digramma in the Philippines displayed complex sexual ontogenies, with all species showing histological evidence of both diandry and functional gonochorism. Reproductive investment in gonadal tissue, and population sex structure, also differed between GBR and Philippine coral reefs. Philippine populations had substantially lower gonado-somatic indices than populations on the GBR. Nonetheless, Philippine populations matured more rapidly and displayed a protracted timing of sex change over a large size and age range. Thus, mature females appeared earlier and persisted later into ontogeny in the Philippines than on GBR reefs. Protracted timing of sex change on Philippine reefs is likely linked to the presence of primary males in the population, which is known to reduce the strength of selection for mature females to undergo sex change and become male. Hypotheses based on social structure of fish populations, environmental factors and evolutionary history were developed to account for the different patterns of sexual ontogeny in the focal wrasses.     

Diversity of social wasps (Hymenoptera: Vespidae) in Cerrado fragments of Uberlândia, Minas Gerais State, Brazil

Studies of species survey are important to know the available natural resources and to get useful information about the ecological characteristics of a specific area. There are not studies, on this issue, related to social wasps, in the Triangulo Mineiro region. The present study describes the diversity of species found in fragments of the cerrado region in Uberlandia, MG, and their temporal distribution. The field study was conducted from October 2003 to September 2004, and 43 samplings were carried out using active searching (24) and point sampling (19) methodologies. Twenty-nine species were found in 10 genera, Polybia and Polistes corresponding to 51.5% of the total listing. Mischocyttarus cerberus styx Richards represented 26.5% of the total individuals recorded by active searching and Agelaia pallipes (Olivier) corresponded to 57.6% by point sampling. Two species had their first record for the state of Minas Gerais: Polybia striata (Fabricius) and M. cerberus styx. The studied fragments presented a high level of diversity (H' = 0.66 to 1.16), a large number of rare species and a few common species. To collect the more common species the point sampling method was satisfactory while active searching was better to collect the rarer species. The active searching methodology was more efficient than point sampling, however some species could only be collected by point sampling. These factors showed that in order to carry out a wasp survey, a combination of different collection methodologies seems to be more appropriate.     

Disruption of gene expression rhythms in mice lacking secretory vesicle proteins IA-2 and IA-2β

Insulinoma-associated protein (IA)-2 and IA-2β are transmembrane proteins involved in neurotransmitter secretion. Mice with targeted disruption of both IA-2 and IA-2β (double-knockout, or DKO mice) have numerous endocrine and physiological disruptions, including disruption of circadian and diurnal rhythms. In the present study, we have assessed the impact of disruption of IA-2 and IA-2β on molecular rhythms in the brain and peripheral oscillators. We used in situ hybridization to assess molecular rhythms in the hypothalamic suprachiasmatic nuclei (SCN) of wild-type (WT) and DKO mice. The results indicate significant disruption of molecular rhythmicity in the SCN, which serves as the central pacemaker regulating circadian behavior. We also used quantitative PCR to assess gene expression rhythms in peripheral tissues of DKO, single-knockout, and WT mice. The results indicate significant attenuation of gene expression rhythms in several peripheral tissues of DKO mice but not in either single knockout. To distinguish whether this reduction in rhythmicity reflects defective oscillatory function in peripheral tissues or lack of entrainment of peripheral tissues, animals were injected with dexamethasone daily for 15 days, and then molecular rhythms were assessed throughout the day after discontinuation of injections. Dexamethasone injections improved gene expression rhythms in liver and heart of DKO mice. These results are consistent with the hypothesis that peripheral tissues of DKO mice have a functioning circadian clockwork, but rhythmicity is greatly reduced in the absence of robust, rhythmic physiological signals originating from the SCN. Thus, IA-2 and IA-2β play an important role in the regulation of circadian rhythms, likely through their participation in neurochemical communication among SCN neurons.