Two-pore channels and disease

Two-pore channels (TPCs) are Ca²⁺-permeable endo-lysosomal ion channels subject to multi-modal regulation. They mediate their physiological effects through releasing Ca²⁺ from acidic organelles in response to cues such as the second messenger, NAADP. Here, we review emerging evidence linking TPCs to disease. We discuss how perturbing both local and global Ca²⁺ changes mediated by TPCs through chemical and/or molecular manipulations can induce or reverse disease phenotypes. We cover evidence from models of Parkinson's disease, non-alcoholic fatty liver disease, Ebola infection, cancer, cardiac dysfunction and diabetes. A need for more drugs targeting TPCs is identified.     

Inhibition of protein synthesis enhances the lytic effects of tumor necrosis factor α and interferon γ in cell lines derived from gynecological malignancies

Summary Few clinical responses have occurred in preliminary studies using the cytokines tumor necrosis factor (TNF) or interferon (IFN) in cancer patients. This may be related to the observation that many malignant cell lines are resistant to lysis by these cytokinesin vitro. Resistance to lysis by TNF or IFN in many cells is controlled by a protein-synthesis-dependent mechanism, such that when protein synthesis is inhibited cells become sensitive to lysis by these cytokines. Because there is some evidence that TNF and IFN act through different lytic mechanisms and are opposed by different resistance mechanisms, we treated a panel of eight cell lines, five derived from human cervical carcinomas (ME-180, MS751, SiHa, HT-3, and C-33A) and three derived from ovarian carcinomas (Caov-3, SK-OV-3, and NIH: OVCAR-3) with both TNF and IFN to determine whether such combination treatment might maximizein vitro cell lysis. Our results showed that pretreatment with IFN followed by exposure to TNF in the presence of protein synthesis inhibitors increased lysis of seven of the eight cell lines above that seen with either TNF or IFN and inhibitors of protein synthesis. Only the cell line C-33A was resistant to lysis by TNF and IFN, when exposed to these agents both alone and in combination with protein synthesis inhibitors. Clinically, combining the cytokines TNF and IFN with protein synthesis inhibitors may maximize thein vivo lytic effects of these cytokines.Supported by American Cancer Society Career Development Award 90-221     

Niche-to-niche migration of bone-marrow-derived cells

During ontogenesis, haematopoietic stem cells (HSCs) relocate between extra-embryonic and embryonic compartments. Similarly, site-specific homing of HSCs is ongoing during adulthood. With the expanding knowledge of HSC physiology, a new paradigm emerges in which HSCs and haematopoietic progenitor cells (HPCs) migrate to defined microenvironments within the bone marrow (BM) and to 'activated' or 'inducible' niches elsewhere. Here, we summarize current understanding of HSC niche characteristics, and the physiological and pathological mechanisms that guide HSC homing both within the BM and to distant niches in the periphery, promoting new vessel growth in tumours and ischaemia. Recent observations suggest that features of the HSC niche might also be recapitulated in pre-metastatic sites. Clusters of BM-derived HPCs promote invasion of disseminating cancer cells. Clear clinical benefits can be foreseen by modulating HSCs and their microenvironments, in promoting tissue regeneration, and inhibiting tumourigenesis and cancer metastasis.     

Sacoglottis gabonensis– a keystone fruit for forest elephants in the Réserve de Faune du Petit Loango, Gabon

This study suggests that the fruits of Sacoglottis gabonensis (Baill.) Urb. (Humiriaceae) are a keystone resource for forest elephants ( Loxodonta cyclotis Matschie) in a coastal rain forest, the Réserve de Faune du Petit Loango, Gabon (now part of Loango National Park). Faecal counts demonstrated that forest elephants used Sacoglottis -dominated forest more when Sacoglottis was abundant and electivity indices suggest that Sacoglottis is a preferred food. The flora of Petit Loango is characterized by the absence of herbaceous vegetation such as Marantaceae and Zingiberaceae, and during the prolonged dry season few fleshy fruits are present other than Sacoglottis fruits, which are produced in a glut during this time. While inter-annual fruiting reliability remains to be confirmed, fruit production in 1998 and high stem density relative to other study sites provide indirect evidence that Sacoglottis fruits are a reliable inter-annual resource at Petit Loango. It is thus proposed that Sacoglottis gabonensis fruits fulfil an important role as a keystone 'fallback' resource for forest elephants during the dry season at Petit Loango.     

Group size, density and biomass of large mammals in the Réserve de Faune du Petit Loango, Gabon

Group size, density and biomass of large‐bodied diurnal mammal species in the Réserve de Faune du Petit Loango, Gabon (now Parc National de Loango) was determined over a 12‐month period using standard line‐transect methods. Petit Loango encompasses a range of distinct habitat types, including coastal scrub, savanna, swamps and disturbed and mature forest. Such intact coastal habitats are increasingly rare on the Central‐West African coastline. Faecal censusing indicated highest forest elephant (Loxodonta africana cyclotis) and buffalo (Syncerus caffer nanus) ecological densities at the extreme coast (2.48 and 1.29 km⁻² respectively), probably reflecting high intensity of use of this habitat. Ape density was comparable with that at other Central African study sites at 1.01 individuals km⁻². Mean total biomass of diurnal primates, elephants and other ungulates over the 20 km² site was 3290 kg km⁻². Forest elephants and red river hogs (Potamochoerus porcus) constituted the bulk of the biomass, at 67% and 14% respectively. Primates made up 5% of the biomass. This is the first estimation of mammal density and biomass over an annual cycle at a Central African coastal site, and provides baseline data for long‐term studies in such habitats and to aid habitat and wildlife management decisions.     

Intergenic enhancers with distinct activities regulate Dlx gene expression in the mesenchyme of the branchial arches

The vertebrate Dlx genes, generally organized as tail-to-tail bigene clusters, are expressed in the branchial arch epithelium and mesenchyme with nested proximodistal expression implicating a code that underlies the fates of jaws. Little is known of the regulatory architecture that is responsible for Dlx gene expression in developing arches. We have identified two distinct cis-acting regulatory sequences, I12a and I56i, in the intergenic regions of the Dlx1/2 and Dlx5/6 clusters that act as enhancers in the arch mesenchyme. LacZ transgene expression containing I12a is restricted to a subset of Dlx-expressing ectomesenchyme in the first arch. The I56i enhancer is active in a broader domain in the first arch mesenchyme. Expression of transgenes containing either the I12a or the I56i enhancers is dependent on the presence of epithelium between the onset of their expression at E9-10 until independence at E11. Both enhancers positively respond to FGF8 and FGF9; however, the responses of the reporter transgenes were limited to their normal domain of expression. BMP4 had a negative effect on expression of both transgenes and counteracted the effects of FGF8. Furthermore, bosentan, a pharmacological inhibitor of Endothelin-1 signaling caused a loss of I56i-lacZ expression in the most distal aspects of the expression domain, corresponding to the area of Dlx-6 expression previously shown to be under the control of Endothelin-1. Thus, the combinatorial branchial arch expression of Dlx genes is achieved through interactions between signaling pathways and intrinsic cellular factors. I56i drives the entire expression of Dlx5/6 in the first arch and contains necessary sequences for regulation by at least three separate pathways, whereas I12a only replicates a small domain of endogenous expression, regulated in part by BMP-4 and FGF-8.     

The effect of different warm-up stretch protocols on 20 meter sprint performance in trained rugby union players

The purpose of this study was to determine the effect of different static and dynamic stretch protocols on 20-m sprint performance. The 97 male rugby union players were assigned randomly to 4 groups: passive static stretch (PSS; n = 28), active dynamic stretch (ADS; n = 22), active static stretch (ASST; n = 24), and static dynamic stretch (SDS; n = 23). All groups performed a standard 10-minute jog warm-up, followed by two 20-m sprints. The 20-m sprints were then repeated after subjects had performed different stretch protocols. The PSS and ASST groups had a significant increase in sprint time (p < or = 0.05), while the ADS group had a significant decrease in sprint time (p < or = 0.05). The decrease in sprint time, observed in the SDS group, was found to be nonsignificant (p > or = 0.05). The decrease in performance for the 2 static stretch groups was attributed to an increase in the musculotendinous unit (MTU) compliance, leading to a decrease in the MTU ability to store elastic energy in its eccentric phase. The reason why the ADS group improved performance is less clear, but could be linked to the rehearsal of specific movement patterns, which may help increase coordination of subsequent movement. It was concluded that static stretching as part of a warm-up may decrease short sprint performance, whereas active dynamic stretching seems to increase 20-m sprint performance.