Autophagy in Trypanosoma brucei: amino acid requirement and regulation during different growth phases.

Autophagy in the protozoan parasite, Trypanosoma brucei, may be involved in differentiation between different life cycle forms and during growth in culture. We have generated multiple parasite cell lines stably expressing green fluorescent protein- or hemagglutinin-tagged forms of the autophagy marker proteins, TbAtg8.1 and TbAtg8.2, in T. brucei procyclic forms to establish a trypanosome system for quick and reliable determination of autophagy under different culture conditions using flow cytometry. We found that starvation-induced autophagy in T. brucei can be inhibited by addition of a single amino acid, histidine, to the incubation buffer. In addition, we show that autophagy is induced when parasites enter stationary growth phase in culture and that their capacity to undergo starvation-induced autophagy decreases with increasing cell density.

Small quantity but large effect — How minor phases control strain localization in upper mantle shear zones

Low viscosity domains such as localized shear zones exert an important control on the geodynamics of the uppermost mantle. Grain size reduction and subsequent strain localization related to a switch from dislocation to diffusion creep is one mechanism to form low viscosity domains. To sustain strain localization, the grain size of mantle minerals needs to be kept small over geological timescales. One way to keep olivine grain sizes small is by pinning of mobile grain boundaries during grain growth by other minerals (second phases). Detailed microstructural studies based on natural samples from three shear zones formed at different geodynamic settings, allowed the derivation of the olivine grain-size dependence on the second-phase content. The polymineralic olivine grain-size evolution with increasing strain is similar in the three shear zones. If the second phases are to pin the mobile olivine grain boundary the phases need to be well mixed before grain growth. We suggest that melt-rock and metamorphic reactions are crucial for the initial phase mixing in mantle rocks. With ongoing deformation and increasing strain, grain boundary sliding combined with mass transfer processes and nucleation of grains promotes phase mixing resulting in fine-grained polymineralic mixtures that deform by diffusion creep. Strain localization due to the presence of volumetrically minor minerals in polymineralic mantle rocks is only important at high strain deformation (ultramylonites) at low temperatures (<~800°C). At smaller strain and stress conditions and/or higher temperatures other parameters like overall energy available to deform a given rock volume, the inheritance of mechanical anisotropies or the presence of water or melts needs to be considered to explain strain localization in the upper mantle.

Climatic change and contemporaneous land-use phases north and south of the Alps 2300 BC to 800 AD

Fluctuations in the Δ14C curve and subsequent gaps of archaeological findings at 800–650 and 400–100 BC in western and central Europe may indicate major climate-driven land-abandonment phases. To address this hypothesis radiocarbon-dated sediments from four lakes in Switzerland were studied palynologically. Pollen analysis indicates contemporaneous phases of forest clearances and of intensified land-use at 1450–1250 BC, 650–450 BC, 50 BC–100 AD and around 700 AD. These land-use expansions coincided with periods of warm climate as recorded by the Alpine dendroclimatic and Greenland oxygen isotope records. Our results suggest that harvest yields would have increased synchronously over wide areas of central and southern Europe during periods of warm and dry climate. Combined interpretation of palaeoecological and archaeological findings suggests that higher food production led to increased human populations. Positive long-term trends in pollen values of Cerealia and Plantago lanceolata indicate that technical innovations during the Bronze and Iron Age (e.g. metal ploughs, scythes, hay production, fertilising methods) gradually increased agricultural productivity. The successful adoption of yield-increasing advances cannot be explained by climatic determinism alone. Combined with archaeological evidence, our results suggest that despite considerable cycles of spatial and demographic reorganisation (repeated land abandonments and expansions, as well as large-scale migrations and population decreases), human societies were able to shift to lower subsistence levels without dramatic ruptures in material culture. However, our data imply that human societies were not able to compensate rapidly for harvest failures when climate deteriorated. Agriculture in marginal areas was abandoned, and spontaneous reforestations took place on abandoned land south and north of the Alps. Only when the climate changed again to drier and warmer conditions did a new wide-spread phase of forest clearances and field extensions occur, allowing the reoccupation of previously abandoned areas. Spatial distribution of cereal cultivation and growth requirements of Cerealia species suggest that increases in precipitation were far more decisive in driving crop failures over central and southern Europe than temperature decreases.

The Jew and Christianity : some phases, ancient and modern, of the Jewish attitude towards Christianity

by Herbert Danby

NRAS and BRAF mutations in primary cutaneous melanoma: A comparison of mutation rates between radial and vertical growth phases in individual tumors

Primary cutaneous melanoma is a cancer arising from melanocytes in the skin. In recent decades the incidence of this malignancy has increased significantly. Mortality rates are high for patients with tumors measuring over a few millimeters in thickness. Response rates to conventional radiation and chemotherapy are very low in patients with metastatic melanoma. New therapies targeting melanoma’s aberrant cell signaling pathways such as the MAP Kinase pathway are being developed. Mutations of NRAS and BRAF genes are quite common in cutaneous melanoma and lead to constitutive activation of the MAP Kinase pathway. This study tests the hypothesis that NRAS and BRAF mutations increase as a tumor progresses from the noninvasive radial growth phase (RGP) to the invasive vertical growth phase (VGP). Laser capture microdissection was used to obtain separate, pure tumor DNA samples from the RGP and VGP of thirty primary cutaneous melanomas. PCR was used to amplify NRAS exon 2 and BRAF exon 15 tumor DNA. The amplified DNA was sequenced and analyzed for mutations. An overall mutation rate of 74% was obtained for the twenty-three melanomas in which there were complete sequence results. With the exception of one melanoma NRAS and BRAF mutations were mutually exclusive. All seven NRAS exon 2 mutations involved codon 61. Three of these melanomas had mutations in both the RGP and VGP. The remaining four tumors were wild type for NRAS exon 2 in the RGP but mutated in the VGP. Of the fifteen BRAF exon 15 mutated melanomas all but one involved codon 600. Twelve of the fifteen BRAF exon 15 mutations were the T1799A type. Nine of the fifteen BRAF mutated tumors had the same mutation in both the RGP and VGP. Five of fifteen melanomas had wild type RGP DNA and BRAF exon 15 mutated VGP DNA. A single melanoma had BRAF exon 15 mutated DNA in the RGP and wild type DNA in the VGP. Overall, these results suggest a trend toward the acquisition of NRAS and BRAF mutations as cutaneous melanomas change from a noninvasive to an invasive, potentially deadly cancer.^