Development of a minigenome cassette for Lettuce necrotic yellows virus: A first step in rescuing a plant cytorhabdovirus

Rhabdoviruses are enveloped negative-sense RNA viruses that have numerous biotechnological applications. However, recovering plant rhabdoviruses from cDNA remains difficult due to technical difficulties such as the need for concurrent in planta expression of the viral genome together with the viral nucleoprotein (N), phosphoprotein (P) and RNA-dependent RNA polymerase (L) and viral genome instability in E. coli. Here, we developed a negative-sense minigenome cassette for Lettuce necrotic yellows virus (LNYV). We introduced introns into the unstable viral ORF and employed Agrobacterium tumefaciens to co-infiltrate Nicotiana with the genes for the N, P, and L proteins together with the minigenome cassette. The minigenome cassette included the Discosoma sp. red fluorescent protein gene (DsRed) cloned in the negative-sense between the viral trailer and leader sequences which were placed between hammerhead and hepatitis delta ribozymes. In planta DsRed expression was demonstrated by western blotting while the appropriate splicing of introduced introns was confirmed by sequencing of RT-PCR product

Sand, salt and water in the Stampriet Basin, Namibia: Calculating unsaturated zone (Kalahari dunefield) recharge using the chloride mass balance approach

Estimating groundwater recharge rates in the Stampriet Basin is important for assessing the sustainability of the groundwater resource both within south-east Namibia, and across the borders of this transboundary resource into Botswana and South Africa. The 65 000 km2 basin contains a multi-layered aquifer system, of up to 9 superimposed aquifers, with a majorupper unconfined and 2 major artesian levels. The majority of water abstraction (66%) is from the unconfined Kalahari Aquifer. This research investigates the direct recharge pathway through the dune-sand cover of the southern Kalahari dunefield which overlies ~80% of the basin. This is undertaken using established chloride mass balance methods within 4unsaturated zone profiles (9 m to 11.5 m depth) from 2 regions of the basin. The results indicate that recent recharge rates are between 7 mm·y-1 to 46 mm·y-1, with profiles representing between 10 years and 30 years of rainfall infiltration. This is the first time that substantial direct (diffuse) recharge has been quantified for the Stampriet Basin, and this part of the southern Kalahari. These 4 profiles indicate potentially similar levels of spatial variability as seen in many other semi-arid, sand-rich unsaturated zones across the world. Further studies using the unsaturated zone should be employed in this region, to better inform our  understanding of recharge mechanisms across the basin. The chloride mass balance technique provides information about the unsaturated zone, and important insights about recharge in various sand-covered regions of the basin for which very little is known, or is possible to determine using conventional water balance approaches. Direct recharge rates estimated in this way constitute a vital part of constraining the water balance of the basin, and particularly of the Kalahari Aquifer from which the majority of groundwater abstraction occurs

Design and implementation of an advanced MQTT broker for distributed pub/sub scenarios

MQTT is one of the most popular communication protocols for Internet of Things applications. Based on a publish/subscribe pattern, it relies on a single broker to exchange messages among clients according to topics of interest. However, such a centralized approach does not scale well and is prone to single point of failure risks, calling for solutions where multiple brokers cooperate together in a distributed fashion. In this paper, we present a complete solution for a distributed MQTT broker systems. We target several functional primitives which are key in such a scenario: broker discovery and failure recovery, overlay tree network creation and message routing. Moreover, we also focus on the case where multiple topics are present in the system. In such a scenario, a single tree-based overlay network connecting the different brokers may not be the most efficient solution. To cope with this issue, we propose a topic-based routing scheme for MQTT distributed brokers. The proposed solution creates multiple overlay networks in the distributed system, each one linking together only the brokers whose connected clients have interest in the same topics. We implement the complete system as an extension of the popular HiveMQ MQTT broker and perform several experiments to test its performance in scenarios characterized by a different publishers/subscribers configurations as well as number of topics existing in the system

A comparative study to investigate the effect of orthodontic treatment on the uniqueness of the human anterior dentition

AIM: The human dentition contains many features which can be used to identify an individual from the dentition or from bite marks created and bite mark evidence may be used to link a suspect to a crime. The aim of this research was to investigate the effect of orthodontic treatment on the uniqueness of the human anterior dentition by comparison of the number of dental shape matches between pre- and post-treatment dental casts for a group of patients who have undergone orthodontic treatment (dental braces) to improve the alignment of their teeth. METHOD: This comparative study utilised pre- and post-orthodontic treatment dental casts from 36 patients. The dental casts were scanned and the anterior 6 teeth landmarked with 24 landmarks in total. The dental casts were divided into 4 groups: pre-orthodontic upper jaw (maxillary) and lower jaw (mandibular) and post-orthodontic maxillary and mandibular. Partial and full Procrustes analyses were undertaken to investigate the similarity between dental casts within each group and whether any of the comparisons were similar enough to be classified as a match. A landmarking repeatability study performed on a set of digitised dental casts determined the error of the landmarking procedure and allowed a proposed match threshold to be established. RESULTS: Orthodontic treatment reduced the uniqueness, and increased the similarity, between dentitions, as evidenced by a reduction in the maximum partial Procrustes distances in the post-orthodontic dental cast groups. None of the dental cast comparisons in the pre- or post-orthodontic maxillary or mandibular groups were classified as a match with the partial Procrustes analysis. However, many false positive matches (between 35 and 61) were identified within the post-orthodontic maxillary and mandibular groups using the full Procrustes analysis. CONCLUSIONS: Orthodontic treatment reduced the uniqueness of the human anterior dentition between different patients. There were no matches identified with the partial Procrustes analysis, but a large number of false positive matches were identified using the full Procrustes analysis. It is therefore proposed that full Procrustes analysis is unsuitable for this type of work and that only partial Procrustes analysis should be utilised

Sustainability of community-based women’s groups: reflections from a participatory intervention for newborn and maternal health in Nepal

Participatory community-based women’s group interventions have been successful in improving maternal and newborn survival. In rural Makwanpur, Nepal, exposure to these Participatory Learning and Action groups resulted in a thirty-percent reduction in neonatal mortality rate and significantly fewer maternal deaths. It is often theorised that participatory approaches are more likely to be sustained than top-down approaches, but this is rarely evaluated after the withdrawal of external support. We sought to understand how participatory learning and action (PLA) groups in Makwanpur fared after the supporting non-governmental organisation withdrew their support as well as factors affecting their sustainability. We used mixed methods, conducting a cross-sectional survey of 239 groups, thirty focus group discussions with group members and thirty key informant interviews within twelve–seventeen months after support was withdrawn. Eighty percent of groups were still active which suggests that PLA groups have a high chance of being sustained over time. Groups were more likely to be sustained if the group had local importance and members continued to acquire new knowledge. However, the participatory nature of the group and local embeddedness were not enough to sustain all groups. They also needed leadership capacity, a unifying activity such as a fund, and a strong belief in the value of their meeting to sustain. These key factors should be considered when seeking to enable sustainability of participatory interventions

Transferability of urban freight transport measures: A case study of Cariacica (Brazil)

This paper presents a case study describing a transferability case study in Cariacica (Brazil), identifying various urban freight planning measures from other parts of the world that might be implemented in Cariacica. The case study developed a light transferability approach, which uses a low level of expert input and data requirements, and is thus appropriate for cities with highly limited financial resources. The case study focussed upon regulatory measures that are under the control of the local authority, and also identified possible barriers and facilitators for their implementation. It was generally considered that the ‘light’ transferability approach had been successful and could be used in many other ‘smaller’ cities. The paper provides insights both for research and transport planning practice (on identifying barriers, and hence solutions, for the successful implementation of urban freight transport measures). Furthermore, the paper provides a number of reflections about the transfer process, paying particular attention to the precise objectives of such exercises and the roles of transfer agents. Finally, conclusions are given which make a further justification of using a ‘light’ approach for policy transfer exercises in the context of the complexity of the urban freight transport system (further ‘complexified’ by transferability issues)

The use of cheese whey for probiotic yogurt production

The objective of this work was to develop a natural yogurt utilizing the resultant whey of a "coalho" cheese production, assaying different processes of pasteurization and powdered milk concentrations. The yogurt submitted to 90 degrees C/5min with 8 and 10% of powdered milk achieved the highest grade on sensorial analysis, with no significant difference ( P > 0,05). Products were characterized as integral, attended the request for protein, and supplied over 15% of the daily feed intake of calcium for children aged 1 to 10, characterizing it as calcium enriched. In both formulations, the pH decreased from 4.6 to 4.2 approximately and the acidity increased from 0.7g to 1.0g lactic acid/100g of product during 28 days of storage. There was no contamination by S. aureus, fecal coliforms, molds and yeasts. The counting of lactic bacteria varied from 5.6x10(9) to 8.0x10(8) CFU/g during shelf life, suggesting the promotion benefits from this probiotic product. It was concluded that whey utilization for yogurt production is a viable alternative to increase economic, nutritional, and functional value.634996100