A note on four nonradioactive labeling systems for dot hybridization detection of potato viruses

Des clones d'ADN complémentaire ont été fabriqués à partir des ARN génomiques des virus S (PVS), X (PVX) et Y (PVY) de la pomme de terre (Solarium tuberosum). Les clones ont été sélectionnés pour leur spécificité par l'hybridation avec divers ARN viraux. Les clones S12 de PVS et X6 de PVX se sont avérés très spécifiques à l'ARN de PVS et PVX respectivement, alors que le clone Y10 de PVY a hybride fortement à l'ARN du PVY et faiblement à l'ARN du PVS. Quatre systèmes commerciaux non radioactifs de marquage des acides nucléiques et de détection ont été comparés entre eux et avec le marquage radioactif traditionnel de la sonde au 32P. La détection colorimétrique de sondes d'ADN marquées à la digoxygénine permet de déceler 1 ng de virions (60 pg d'ARN), soit une sensibilité du même ordre que l'autoradiographie avec des sondes marquées au phosphore radioactif. Les sondes sulfonées, biotinylées et marquées à la peroxydase ont été moins sensibles en permettant la détection de 600 pg d'ARN viral.Complementary DNA clones of genomic RNAs of potato (Solarium tuberosum) viruses S (PVS), X (PVX) and Y (PVY) were produced and tested for their capacity to hybridize with various plant virus RNAs. PVS clone S12 and PVX clone X6 were found to be very specifie to PVS and PVX RNA respectively, whereas PVY clone Y10 strongly hybridized with PVY RNA and weakly with PVS RNA. Four commercial, nonradioactive Systems of nucleic acid labeling and detection were compared to the usual 32P-labeled probe using dot hybridization experiments. Colorimetric detection of digoxigenin-labeled DNA probes gave a level of sensitivity of 1 ng of virions (60 pg of RNA), similar to autoradiography of 32P-labeled probes. Sulfonated, biotinylated and peroxidase-labeled probes were slightly less sensitive, allowing detection of 600 pg of viral RNA

Peran Assistant Colorist dalam Proses Color Grading di ABOVE Space

Ketertarikan penulis terhadap color grading berawal dari pengalaman penulis selama berkuliah di program studi film Universitas Multimedia Nusantara, penulis sering mengerjakan proyek film pendek dan berperan sebagai sinematografer yang juga merangkap menjadi colorist untuk proyek yang sama. Hal ini membuat penulis memutuskan untuk melakukan proses magang pada divisi color grading di ABOVE Space dengan tujuan mengetahui lebih dalam tentang color grading dan bagaimana pekerjaan tersebut dilakukan dalam industri profesional. Dalam program magang ini, penulis berperan sebagai assistant colorist yang memiliki tanggung jawab untuk membantu pekerjaan para colorist di ABOVE Space. Pada laporan ini, penulis membahas tentang tahapan-tahapan kerja yang dilakukan assistant colorist, yaitu conforming, menghadiri sesi color grading bersama client di studio, applying, finishing, dan rendering. Selama melakukan kerja magang sebagai assistant colorist di ABOVE Space, penulis banyak belajar tentang hal- hal yang teknis maupun non teknis terkait color grading. Dari segi teknis, penulis dapat mengetahui bagaimana workflow untuk color grading dalam dunia professional, dimulai dari tahap menerima materi untuk color grading sampai pada tahap rendering hasil atau output color grading. Penulis juga dapat mempelajari cara para colorist berkomunikasi langsung dengan client mereka, seperti sutradara, sinematografer, agency, dan lain sebagainya. Sebagai colorist, tidak hanya mahir dalam teknis color grading, penulis juga mempelajari bahwa penting untuk berpikir cepat dalam menemukan solusi-solusi dari kendala yang di hadapi

Estimasi Suhu Permukaan Tanah Menggunakan Metode Algoritma Mono Window di Kabupaten Lamongan

Perubahan alih fungsi lahan merupakan salah satu fenomena yang banyak terjadi pada kabupaten yang berpotensi untuk pengembangan wilayah tersebut, salah satu area yang berpotensi untuk menjadi Kawasan Ekonomi Kreatif adalah Kabupaten Lamongan. Penelitian ini bertujuan untuk estimasi perubahan suhu permukaan tanah di Kabupaten Lamongan dengan memanfaatkan citra satelit Landsat 9. Citra satelit Landsat 9 diolah berdasarkan algoritma suhu untuk estimasi perhitungan suhu. Satelit Landsat 9 diluncurkan pada tahun 2021 dan merupakan satelit pengganti dari satelit Landsat 8. Satelit Landsat 9 memiliki 9 (Sembilan) band spectral dan 2 (dua) band termal. Pengolahan estimasi suhu permukaan tanah menggunakan band 4, band 5, dan band 10. Algoritma suhu permukaan tanah yang digunakan adaalah algoritma mono window. Dilakukan proses mosaic pada path 118 raw 65 dan path 119 raw 65. Hasil penelitian menunjukkan bahwa wilayah Kabupaten Lamongan memiliki tingkat kehijauan dengan rentang nilai -0,611 sampai 0,5 yang didominasi oleh tingkat kerapatan sedang. Hasil estimasi suhu permukaan tanah di Kabupaten Lamongan didominasi oleh suhu dengan rentang 23°C – 26°C dengan suhu tertinggi 29°C – 33°C. Kata kunci: Citra, Landsat 9, Suhu permukaan tana

Revolatilisation of soil-accumulated pollutants triggered by the summer monsoon in India

Persistent organic pollutants that have accumulated in soils can be remobilised by volatilisation in response to chemical equilibrium with the atmosphere. Clean air masses from the Indian Ocean, advected with the onset of the summer monsoon, are found to reduce concentrations of hexachlorocyclohexane (HCH), dichlorodiphenyltrichloroethane (DDT) and its derivatives, endosulfan and polychlorinated biphenyls (PCBs) in air at a mountain site (all in the range 5–20&thinsp;pg&thinsp;m−3) by 77&thinsp;%, 70&thinsp;%, 82&thinsp;% and 45&thinsp;%, respectively. The analysis of fugacities in soil and air suggest that the arrival of summer monsoon triggers net volatilisation or enhances ongoing revolatilisation of the now-banned chemicals HCH and PCBs from background soils in southern India. The response of the air–soil exchange was modelled using a regional air pollution model, WRF-Chem PAH/POP. The results suggest that the air is increasingly polluted during transport by the south-westerly monsoon winds across the subcontinent. Using a multidecadal multimedia mass balance model, it is found that air–surface exchange of HCH and DDT have declined since the ban of these substances from agriculture, but remobilisation of higher chlorinated PCBs may have reached a historical high, 40 years after peak emission.</p

Nongenomic oestrogen signalling in oestrogen receptor negative breast cancer cells: a role for the angiotensin II receptor AT1

INTRODUCTION: Oestrogens can mediate some of their cell survival properties through a nongenomic mechanism that involves the mitogen-activated protein kinase (MAPK) pathway. The mechanism of this rapid signalling and its dependence on a membrane bound oestrogen receptor (ER), however, remains controversial. The role of G-protein-coupled receptor and epidermal growth factor (EGF) receptor in an ER-independent signalling pathway modulated by oestrogen was investigated. METHODS: ER-positive and ER-negative breast cancer cell lines (MCF-7 and SKBR3) and primary breast cancer cell cultures were used in this study. Cell proliferation was assessed using standard MTT assays. Protein and cAMP levels were detected by Western blotting and ELISA, respectively. Antigen localization was performed by immunocytochemistry, immunohistochemistry and immunofluorescence. Protein knockdown was achieved using small interfering RNA technologies. RESULTS: EGF and oestrogen, alone and in combination, induced cell proliferation and phosphorylation of MAPK proteins Raf and ERK (extracellular signal regulated kinase)1/2 in both ER-negative SKBR3 and ER-positive MCF-7 human breast cancer cell lines. Increased Raf phosphorylation was also observed in primary human breast cultures derived from ER-positive and ER-negative breast tumours. Oestrogen induced an increase in intracellular cAMP in ER-negative SKBR3 human breast cancer cells. Oestrogen-mediated cell growth and phosphorylation of MAPK was modified by the EGF receptor antagonist AG1478, the G-protein antagonist pertussis toxin, and the angiotensin II receptor antagonist saralasin. Knockdown of angiotensin II type 1 receptor (AT1) protein expression with small interfering RNA attenuated oestrogen-induced Raf phosphorylation in ER-negative cells. AT1 receptor was found to be expressed in the cell membrane of breast tumour epithelial cells. CONCLUSION: These findings provide evidence that, in breast cancer cells, oestrogen can signal through AT1 to activate early cell survival mechanisms in an ER-independent manner

Scrub typhus ecology: a systematic review of Orientia in vectors and hosts

Abstract Scrub typhus, caused by Orientia tsutsugamushi, is an important and neglected vector-borne zoonotic disease with an expanding known distribution. The ecology of the disease is complex and poorly understood, impairing discussion of public health interventions. To highlight what we know and the themes of our ignorance, we conducted a systematic review of all studies investigating the pathogen in vectors and non-human hosts. A total of 276 articles in 7 languages were included, with 793 study sites across 30 countries. There was no time restriction for article inclusion, with the oldest published in 1924. Seventy-six potential vector species and 234 vertebrate host species were tested, accounting for over one million trombiculid mites (‘chiggers’) and 83,000 vertebrates. The proportion of O. tsutsugamushi positivity was recorded for different categories of laboratory test and host species. Vector and host collection sites were geocoded and mapped. Ecological data associated with these sites were summarised. A further 145 articles encompassing general themes of scrub typhus ecology were reviewed. These topics range from the life-cycle to transmission, habitats, seasonality and human risks. Important gaps in our understanding are highlighted together with possible tools to begin to unravel these. Many of the data reported are highly variable and inconsistent and minimum data reporting standards are proposed. With more recent reports of human Orientia sp. infection in the Middle East and South America and enormous advances in research technology over recent decades, this comprehensive review provides a detailed summary of work investigating this pathogen in vectors and non-human hosts and updates current understanding of the complex ecology of scrub typhus. A better understanding of scrub typhus ecology has important relevance to ongoing research into improving diagnostics, developing vaccines and identifying useful public health interventions to reduce the burden of the disease.</jats:p

Nonrandom Distribution of Vector Ticks (Dermacentor variabilis) Infected by Francisella tularensis

The island of Martha's Vineyard, Massachusetts, is the site of a sustained outbreak of tularemia due to Francisella tularensis tularensis. Dog ticks, Dermacentor variabilis, appear to be critical in the perpetuation of the agent there. Tularemia has long been characterized as an agent of natural focality, stably persisting in characteristic sites of transmission, but this suggestion has never been rigorously tested. Accordingly, we sought to identify a natural focus of transmission of the agent of tularemia by mapping the distribution of PCR-positive ticks. From 2004 to 2007, questing D. variabilis were collected from 85 individual waypoints along a 1.5 km transect in a field site on Martha's Vineyard. The positions of PCR-positive ticks were then mapped using ArcGIS. Cluster analysis identified an area approximately 290 meters in diameter, 9 waypoints, that was significantly more likely to yield PCR-positive ticks (relative risk 3.3, P = 0.001) than the rest of the field site. Genotyping of F. tularensis using variable number tandem repeat (VNTR) analysis on PCR-positive ticks yielded 13 different haplotypes, the vast majority of which was one dominant haplotype. Positive ticks collected in the cluster were 3.4 times (relative risk = 3.4, P<0.0001) more likely to have an uncommon haplotype than those collected elsewhere from the transect. We conclude that we have identified a microfocus where the agent of tularemia stably perpetuates and that this area is where genetic diversity is generated

Enabling the freight traffic controller for collaborative multi-drop urban logistics: practical and theoretical challenges

There is increasing interest in how horizontal collaboration between parcel carriers might help alleviate problems associated with last-mile logistics in congested urban centers. Through a detailed review of the literature on parcel logistics pertaining to collaboration, along with practical insights from carriers operating in the United Kingdom, this paper examines the challenges that will be faced in optimizing multicarrier, multidrop collection, and delivery schedules. A “freight traffic controller” (FTC) concept is proposed. The FTC would be a trusted third party, assigned to equitably manage the work allocation between collaborating carriers and the passage of vehicles over the last mile when joint benefits to the parties could be achieved. Creating this FTC concept required a combinatorial optimization approach for evaluation of the many combinations of hub locations, network configuration, and routing options for vehicle or walking to find the true value of each potential collaboration. At the same time, the traffic, social, and environmental impacts of these activities had to be considered. Cooperative game theory is a way to investigate the formation of collaborations (or coalitions), and the analysis used in this study identified a significant shortfall in current applications of this theory to last-mile parcel logistics. Application of theory to urban freight logistics has, thus far, failed to account for critical concerns including (a) the mismatch of vehicle parking locations relative to actual delivery addresses; (b) the combination of deliveries with collections, requests for the latter often being received in real time during the round; and (c) the variability in travel times and route options attributable to traffic and road network conditions

Peatland Initiation, Carbon Accumulation, and 2 ka Depth in the James Bay Lowland and Adjacent Regions

Copyright © 2014 University of Colorado at Boulder, Institute of Arctic and Alpine ResearchPeatlands surrounding Hudson and James Bays form the second largest peatland complex in the world and contain major stores of soil carbon (C). This study utilized a transect of eight ombrotrophic peat cores from remote regions of central and northern Ontario to quantify the magnitude and rate of C accumulation since peatland initiation and for the past 2000 calendar years before present (2 ka). These new data were supplemented by 17 millennially resolved chronologies from a literature review covering the Boreal Shield, Hudson Plains, and Taiga Shield bordering Hudson and James Bays. Peatlands initiated in central and northern Ontario by 7.8 ka following deglaciation and isostatic emergence of northern areas to above sea level. Total C accumulated since inception averaged 109.7 ± (std. dev.) 36.2 kg C m–2. Approximately 40% of total soil C has accumulated since 2 ka at an average apparent rate of 20.2 ± 6.9 g C m–2 yr–1. The 2 ka depths correlate significantly and positively with modern gridded climate estimates for mean annual precipitation, mean annual air temperature, growing degree-days > 0 °C, and photosynthetically active radiation integrated over days > 0 °C. There are significantly shallower depths in permafrost peatlands. Vertical peat accumulation was likely constrained by temperature, growing season length, and photosynthetically active radiation over the last 2 ka in the Hudson Bay Lowlands and surrounding regions.US National Science Foundatio