Haploinsufficiency of a spliceosomal GTPase encoded by EFTUD2 causes mandibulofacial dysostosis with microcephaly

Mandibulofacial dysostosis with microcephaly (MFDM) is a rare sporadic syndrome comprising craniofacial malformations, microcephaly, developmental delay, and a recognizable dysmorphic appearance. Major sequelae, including choanal atresia, sensorineural hearing loss, and cleft palate, each occur in a significant proportion of affected individuals. We present detailed clinical findings in 12 unrelated individuals with MFDM; these 12 individuals compose the largest reported cohort to date. To define the etiology of MFDM, we employed whole-exome sequencing of four unrelated affected individuals and identified heterozygous mutations or deletions of EFTUD2 in all four. Validation studies of eight additional individuals with MFDM demonstrated causative EFTUD2 mutations in all affected individuals tested. A range of EFTUD2-mutation types, including null alleles and frameshifts, is seen in MFDM, consistent with haploinsufficiency; segregation is de novo in all cases assessed to date. U5-116kD, the protein encoded by EFTUD2, is a highly conserved spliceosomal GTPase with a central regulatory role in catalytic splicing and post-splicing-complex disassembly. MFDM is the first multiple-malformation syndrome attributed to a defect of the major spliceosome. Our findings significantly extend the range of reported spliceosomal phenotypes in humans and pave the way for further investigation in related conditions such as Treacher Collins syndrome.     

Traps and attractants for wood-boring insects in ponderosa pine stands in the Black Hills, South Dakota

Recent large-scale wildfires have increased populations of wood-boring insects in the Black Hills of South Dakota. Because little is known about possible impacts of wood-boring insects in the Black Hills, land managers are interested in developing monitoring techniques such as flight trapping with semiochemical baits. Two trap designs and four semiochemical attractants were tested in a recently burned ponderosa pine, Pinus ponderosa Dougl. ex Laws., forest in the Black Hills. Modified panel and funnel traps were tested in combination with the attractants, which included a woodborer standard (ethanol and alpha-pinene), standard plus 3-carene, standard plus ipsenol, and standard plus ipsdienol. We found that funnel traps were equally efficient or more efficient in capturing wood-boring insects than modified panel traps. Trap catches of cerambycids increased when we added the Ips spp. pheromone components (ipsenol or ipsdienol) or the host monoterpene (3-carene) to the woodborer standard. During the summers of 2003 and 2004, 18 cerambycid, 14 buprestid, and five siricid species were collected. One species of cerambycid, Monochamus clamator (LeConte), composed 49 and 40% of the 2003 and 2004 trap catches, respectively. Two other cerambycids, Acanthocinus obliquus (LeConte) and Acmaeops proteus (Kirby), also were frequently collected. Flight trap data indicated that some species were present throughout the summer, whereas others were caught only at the beginning or end of the summer.     

Dark starvation and plant metabolism

Summary The fixation pattern of radioactive labelled photosynthetic intermediates was followed under steady state conditions during prolonged dark starvation of spinach plants (Spinacia oleracea L.). It is suggested that the considerable increase of radioactive dihydroxyacetonephosphate is correlated with a specific leakage of the outer chloroplast envelope induced by dark starvation. The primary fixation product, phosphoglyceric acid, followed the same decreasing tendency as observed for the net CO₂ fixation. In contrast, the relative label in other intermediates is the same as in the controls. When after several days of dark starvation the plants were again transferred into light, a regeneration of the CO₂ fixation accompanied by the appearance of a normal fixation pattern was observed. Since the regeneration was prevented by the addition of lincomycin, the net increase is considered to be due to a new protein synthesis rather than a reactivation.Abbreviations GAPDH glyceraldehyde phosphate dehydrogenase (E.C. 1.2.1.13) - DHAP dihydroxyacetonphosphate - FDP fructose 1,6-diphosphate - glol glycolic acid - PGA 3-phosphoglyceric acid - S-D-P sugar diphosphates - S-M-P sugar monophosphates Part I: Postius and Jacobi (1971).     

Factors Related to the Recovery of Subalpine Woodland on Mauna Kea,Hawaii1

We measured mature tree and sapling density, tree associations, crown size, age structure, recovery from ungulate browsing, and grass cover at four study sites in two types of subalpine woodland on Mauna Kea volcano, island of Hawaii. Beginning in 1981, introduced ungulates were reduced in number to allow regeneration of Sophora chrysophylla (mamane) in habitat supporting the endangered Hawaiian finch, Loxioides bailleui (palila). We found Sophora regeneration at all four study sites, but regeneration was higher in mixed species woodland with codominant Myoporum sandwicense (naio) than in areas where Sophora dominated. Regeneration of Myoporum was uniformly very low in comparison. Invasive grass cover, which suppresses Sophora germination, was highest in mid-elevation woodland where Sophora dominated. The distribution of mature and sapling Sophora were both related to study site, reflecting previous ungulate browsing and uneven recovery due to grasses. Densities of Sophora snags were not different among any of the sites, suggesting a more even distribution in the past. Selective browsing before ungulate reduction may have favored Myoporum over Sophora, leading to high densities of mature Myoporum in codominant woodland. After ungulate reduction, however, we found no pattern of competitive inhibition by Myoporum on regeneration of Sophora. Reduction of Myoporum is not likely to enhance habitat for Loxioides as much as supplemental plantings of Sophora, glass control, and continued ungulate eradication. Mid-elevation Sophora woodland areas, where Loxioides forage and nest in high densities, would benefit the most from these management actions.     

Sonic hedgehog promotes proliferation and differentiation of adult muscle cells: Involvement of MAPK/ERK and PI3K/Akt pathways

Sonic hedgehog (Shh) has been reported to act as a mitogen and survival factor for muscle satellite cells. However, its role in their differentiation remains ambiguous. Here, we provide evidence that Shh promotes the proliferation and differentiation of primary cultures of chicken adult myoblasts (also termed satellite cells) and mouse myogenic C2 cells. These effects are reversed by cyclopamine, a specific chemical inhibitor of the Shh pathway. In addition, we show that Shh and its downstream molecules are expressed in adult myoblast cultures and localize adjacent to Pax7 in muscle sections. These gene expressions are regulated during postnatal muscle growth in chicks. Most importantly, we report that Shh induces MAPK/ERK and phosphoinositide 3-kinase (PI3K)-dependent Akt phosphorylation and that activation of both signaling pathways is essential for Shh's signaling in muscle cells. However, the effect of Shh on Akt phosphorylation is more robust than that on MAPK/ERK, and data suggest that Shh influences these pathways in a manner similar to IGF-I. By exploiting specific chemical inhibitors of the MAPK/ERK and PI3K/Akt signaling pathways, UO126 and Ly294002, respectively, we demonstrate that Shh-induced Akt phosphorylation, but not that of MAPK/ERK, is required for its promotive effects on muscle cell proliferation and differentiation. Taken together, we suggest that Shh acts in an autocrinic manner in adult myoblasts, and provide first evidence of a role for PI3K/Akt in Shh signaling during myoblast differentiation.     

Mitochondria-mediated ATP depletion by anti-cancer agents of the jasmonate family

Jasmonates are plant stress hormones that induce suppression of proliferation and death in cancer cells, while being selectively inactive towards non-transformed cells. Jasmonates can overcome apoptotic blocks and exert cytotoxic effects on drug-resistant cells expressing p53 mutations. Jasmonates induce a rapid depletion of ATP in cancer cells. Indeed, this steep drop occurs when no signs of cell death are detectable yet. Experiments using modulators of ATP synthesis via glycolysis or oxidative phosphorylation suggest that the latter is the pathway suppressed by jasmonates. Consequently, the direct effects of jasmonates on mitochondria were evaluated. Jasmonates induced cytochrome c release and swelling in mitochondria isolated from cancer cells but not from normal ones. Thus, the selectivity of jasmonates against cancer cells is rooted at the mitochondrial level, and probably exploits differences between mitochondria from normal versus cancer cells. These findings position jasmonates as promising anti-cancer drugs acting via energetic depletion in neoplastic cells.