An improved method using densitometry for evaluating severity of laser photocoagulation induced CNV

Laser photocoagulation induced choroidal neovascularization currently is the most effective model available for the study of this disease in terms of efficacy of new drugs and therapies. Previously, evaluating the extent of choroidal neovascularization using this model was time- consuming and required the use of experienced personnel. We describe a new method for simple and rapid evaluation of laser induced choroidal neovascularization using densitometry. Fluorescein angiograms of a laser photocoagulated rat eye were scanned into a computer. Densitometry software subsequently was used to calculate the severity of the laser lesions. The densitometry method proved effective for calculating the extent of laser induced choroidal neovascularization. In addition, this method was more rapid than visual evaluations and less likely to produce errors.     

Possible role of a cAMP-dependent phosphorylation in the calcium release mediated by inositol 1,4,5-trisphosphate in human platelet membrane vesicles

The addition of inositol 1,4,5-trisphosphate (IP3) to a 45Ca-preloaded human platelet membrane fraction (dense tubular system) induced a transient release of Ca2+. When the vesicle fraction was loaded with 45Ca2+ to isotopic equilibrium in the presence of the catalytic subunit of the cAMP-dependent protein kinase, the level of Ca2+ uptake was increased and the subsequent IP3-induced Ca2+ release was enhanced. The stimulation was observed regardless of the IP3 concentration used, and was maximal with an enzyme concentration of 5 micrograms/ml. The addition of the protein kinase inhibitor prevented the stimulatory effect of the catalytic subunit on IP3-induced calcium release, and also abolished the calcium release detected in the absence of added enzyme. It is concluded that a cAMP-dependent protein phosphorylation may be involved in the regulation of the IP3-induced Ca2+ release in human platelets.     

Deconstructing Ixodes ricinus: a partial matrix model allowing mapping of tick development,mortality and activity rates

A stage‐structured Leslie matrix model of a partial, discrete population of Ixodes ricinus (Linnaeus) (Ixodida: Ixodidae) ticks was developed to elucidate the impact of climate trends on the distribution and phenology of this species in the western Palaearctic. The model calculates development and mortality rates for each instar and evaluates recruitment rates based on the development of the tick population. The model captures the changes in development and mortality rates, providing a coherent index of performance correlated with the tick's geographic range. Maximum development rates are recorded for latitudes south of 36 °N and are spatially correlated with sites of maximum temperature, highest saturation deficit and highest mortality. The maximum available developmental time (the total annual time during which temperature allows development) for I. ricinus in the western Palaearctic is < 45% of the total year. North of 60 °N, available developmental time decreases sharply to only 15% of the year. The latitudinal boundary at which survival rates sharply drop is 43–46 °N, clearly delimiting the classically recognized extent of the main tick populations. The pattern of activity for larval–nymphal synchrony shows a clear west–east pattern. The model demonstrates the impact of climate according to tick stage and geographic location, and provides a practical framework for testing how the tick's lifecycle is affected by climate change.     

Long‐proboscid brachyceran flies in Cretaceous amber (Diptera: Stratiomyomorpha: Zhangsolvidae)

The monophyletic family Zhangsolvidae comprises stout‐bodied brachyceran flies with a long proboscis and occurring only in the Cretaceous, originally known in shale from the Early Cretaceous Laiyang Formation (Fm.) in China (Zhangsolva Nagatomi & Yang), subsequently from limestones of the Early Cretaceous Crato Fm. of Brazil. Cratomyoides Wilkommen is synonymized with Cratomyia Mazzarolo & Amorim, both from the Crato Fm.; Cratomyiidae is synonymized with Zhangsolvidae. Two genera and three species of Zhangsolvidae are described: Buccinatormyia magnifica Arillo, Peñalver & Pérez‐de la Fuente, gen. et sp.n. and B. soplaensis Arillo, Peñalver & Pérez‐de la Fuente, sp.n. , in Albian amber from Las Peñosas Fm. in Spain; and Linguatormyia teletacta Grimaldi, gen. et sp.n. , in Upper Albian–Lower Cenomanian amber from Hukawng Valley in Myanmar. Buccinatormyia soplaensis and Linguatormyia teletacta are unique among all Brachycera, extant or extinct, by their remarkably long, flagellate antennae, about 1.6× the body length in the latter species. A phylogenetic analysis of 52 morphological characters for 35 taxa is presented, 11 taxa being Cretaceous species, which supports placement of the family within Stratiomyomorpha, although not to any particular family within the infraorder. This published work has been registered in Zoobank, http://zoobank.org/urn:lsid:zoobank.org:pub:F32CF887‐7C37‐45D5‐BD6B‐135FE9B729A7 .     

Distribution of streptothricin acetyltransferase encodi determinants among environmental bacteria

In this paper we report about a screening for streptothricin- (St)-resistant phenotypes and genotypes among environmental bacteria from a St virgin area. St-resistant bacteria were isolated from river water, sewage, manure and soil by selective plating. The resistance quotient was typical of an area without selective pressure. The occurrence of streptothricin acetyltransferase-encoding determinants and their localization on a Tn7-like transposon was tested by the application of a set of gene probes. Sat genes could be detected in 22.5% of the tested St-resistant bacteria but in 100% of the checked Enterobacteriaceae. However, we could not detect sat genes in St-resistant bacteria from soil samples. Surprisingly the sat genes were found to be located on conjugative or mobilizable plasmids for a rather high number of strains. The determined plasmid species and their restriction patterns showed a high degree of similarities to those observed from an area of strong selective pressure.     

The invasive predator <Emphasis Type="Italic">Bythotrephes</Emphasis> induces changes in the vertical distribution of native copepods in Lake Michigan

Invasive predators can have large negative effects on native prey populations. The susceptibility of native prey to invasive predators may depend on their ability to respond behaviorally to the presence of these non-native predators. In a field survey conducted in Lake Michigan over several years, we found that high densities of the invasive predatory cladoceran Bythotrephes were correlated with lower vertical distributions of some species and age classes of native copepods; moving from inhabiting primarily the epiliminion at low Bythotrephes density to primarily the hypolimnion at high Bythotrephes density. Five groups showed this pattern; diaptomid copepodites, adult cyclopoids, Diacyclops thomasi, and the adult diaptomids Leptodiaptomus ashlandi and L. minutus. In contrast, Bythotrephes density was not correlated with the vertical distribution of copepod nauplii and adult L. sicilis. Laboratory experiments suggest that the changes in the vertical distribution in the field at high Bythothrephes are due to an inducible, plastic response to predation threat from Bythotrephes signaled by water-borne cues. Species that were lower in the field at high Bythotrephes densities responded behaviorally to water-borne cues from Bythotrephes by moving to lower levels of experimental water columns. These species included D. thomasi and L. minutus, with L. ashlandi displaying a non-significant trend in the same direction. In contrast, L. sicilis, which was not correlated with Bythotrephes density in the field, was unaffected by the water-borne cues. Differences in vertical distribution shifts among these native copepod species and life-history stages are likely due to species-specific differences in spatial overlap with Bythotrephes and their relative ability to migrate large distances or employ alternative avoidance strategies. The varied responses exhibited among the copepod groups likely alter their interactions with each other, their resources and other predators, thus revealing the complex effects Bythotrephes can have on invaded communities.     

FUS contributes to mTOR-dependent inhibition of translation

The amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD)–linked RNA-binding protein called FUS (fused in sarcoma) has been implicated in several aspects of RNA regulation, including mRNA translation. The mechanism by which FUS affects the translation of polyribosomes has not been established. Here we show that FUS can associate with stalled polyribosomes and that this association is sensitive to mTOR (mammalian target of rapamycin) kinase activity. Specifically, we show that FUS association with polyribosomes is increased by Torin1 treatment or when cells are cultured in nutrient-deficient media, but not when cells are treated with rapamycin, the allosteric inhibitor of mTORC1. Moreover, we report that FUS is necessary for efficient stalling of translation because deficient cells are refractory to the inhibition of mTOR-dependent signaling by Torin1. We also show that ALS-linked FUS mutants R521G and P525L associate abundantly with polyribosomes and decrease global protein synthesis. Importantly, the inhibitory effect on translation by FUS is impaired by mutations that reduce its RNA-binding affinity. These findings demonstrate that FUS is an important RNA-binding protein that mediates translational repression through mTOR-dependent signaling and that ALS-linked FUS mutants can cause a toxic gain of function in the cytoplasm by repressing the translation of mRNA at polyribosomes.     

Resolving UV Bioactinometric Results with Intensity and Time Distributions

A UV reactor with an annular design, a total liquid volume of 460[emsp4 ]ml, and outfitted with a single lamp with 1690[emsp4 ]mW of germicidal power was tested. Coliphage MS2 was used as a bioactinometer to measure the UV dose at a flow rate of 56.7[emsp4 ]ml/sec in water with a very low absorbance. The Beers Law coefficient was A₁₀0.003. The measured dose (MS2 bioactinometry) was 35.2±1.1[emsp4 ]mW-sec/cm².A retention time distribution was generated with a dye tracer study. The reactor was modeled as if flow was confined to ten equal volume paths existing as concentric rings around the lamp. The UV intensity along each path (ith intensity) was calculated to generate a simulated distribution of UV intensity in the reactor. The retention time distribution was subdivided to estimate the retention time associated with each decile jth time) of the total flow.Seven methods of associating the ith intensity with the jth retention time were used to produce simulated dose distributions for the reactor. The average UV dose for each distribution was calculated as the average of the products of I and t (AP protocol) and by the apparent survival (AS protocol), in which the predicted survival along each path was averaged to back-calculate dose from the reference batch inactivation curve. The average dose predicted assuming that time and intensity were independent was 51.5[emsp4 ]mW-sec/cm² based on the arithmetic average (AP protocol). Using the apparent survival method, the predicted dose for the independent distribution (I independent of t) was 36.4[emsp4 ]mW-sec/cm². Three methods of developing dependent structure between time and intensity were tested. In the best possible case for stratified flow (I negatively correlated with t) the calculated (AS) intensity was 46.3[emsp4 ]mW-sec/cm^2. In the worst case for stratified flow (I positively correlated with t) the AS intensity was 32.0[emsp4 ]mW-sec/cm². In a rational case where flows were assumed to be distributed parabolically (low flow at the wall and at the lamp) produced an AS intensity of 37.7[emsp4 ]mW-sec/cm². When either time or intensity was averaged, while the other variable was allowed to keep its distribution, the (AS) dose (time averaged 43.3[emsp4 ]mW-sec/cm², intensity averaged 41.0[emsp4 ]mW-sec/cm²), yielded a poor prediction compared to the measured value.The errors associated with averaging time, intensity, or both, far outweigh the errors associated with choosing a rational distribution or an independent distribution of time and intensity in the prediction. This observation is generally true whenever an organism is exposed to UV light in a flow through reactor such that the range of doses is within the portion of the inactivation curve exhibiting strong exponential decay.