Sesbania Mosaic Virus (SeMV) Infectious Clone: Possible Mechanism of 3 ` and 5 ` End Repair and Role of Polyprotein Processing in Viral Replication

Sesbania mosaic virus (SeMV) is a positive stranded RNA virus belonging to the genus Sobemovirus. Construction of an infectious clone is an essential step for deciphering the virus gene functions in vivo. Using Agrobacterium based transient expression system we show that SeMV icDNA is infectious on Sesbania grandiflora and Cyamopsis tetragonoloba plants. The efficiency of icDNA infection was found to be significantly high on Cyamopsis plants when compared to that on Sesbania grandiflora. The coat protein could be detected within 6 days post infiltration in the infiltrated leaves. Different species of viral RNA (double stranded and single stranded genomic and subgenomic RNA) could be detected upon northern analysis, suggesting that complete replication had taken place. Based on the analysis of the sequences at the genomic termini of progeny RNA from SeMV icDNA infiltrated leaves and those of its 3' and 5' terminal deletion mutants, we propose a possible mechanism for 3' and 5' end repair in vivo. Mutation of the cleavage sites in the polyproteins encoded by ORF 2 resulted in complete loss of infection by the icDNA, suggesting the importance of correct polyprotein processing at all the four cleavage sites for viral replication. Complementation analysis suggested that ORF 2 gene products can act in trans. However, the trans acting ability of ORF 2 gene products was abolished upon deletion of the N-terminal hydrophobic domain of polyprotein 2a and 2ab, suggesting that these products necessarily function at the replication site, where they are anchored to membranes.

Assessment of post-harvest losses in Nigeria fishery: the Kainji Lake model

The paper provides a description of a methodology used for quantitative assessment of post harvest losses in the Kainji Lake Fishery (Nigeria). The sample population was made up of 314 fisherfolk, 115 processors, 125 fish buyers and 111 fish sellers. For the determination of handling losses, 24,839 fishes weighing 2,389.31 kg belonging to 43 species were examined of which 10% by number and 9% by weight deteriorated at checking and 4% by number and 3% by weight at landing. Processing losses recorded 22% by number and 16% by weight deteriorated prior to and during smoking with the traditional 'Banda' kiln. During marketing, 16% of fish sold had deteriorated and 6% by weight of fish bought also deteriorated, mainly due to insect infestation during storage. Based on the 1995 yield estimate for Kainji Lake fishery, approximately 1000 tons of fish estimated at 80 million Naira were lost during handling alone. This figure would be much higher if the level of losses during processing and marketing are included. This assessment technique is recommended for use in obtaining quantifiable data on post harvest losses from other water bodies in Nigeria

Fish quality and processing in Malawi: responding to challenges through institutional capacity building

Fish processing and quality control in Malawi are still poorly developed. Traditional fish processing methods are widely employed resulting in considerable post-harvest losses. One of the major challenges to steady and sustainable development in fish processing and quality management is the lack of adequately trained personel. This is directly reflected in poor institutional capacity. This project analyses the situation in fish processing and quality management in Malawi to identify gaps that require improvement. Specifically, the project assesses the role of training institutions in Malawi in capacity building for fish processing and quality management. The institution under discussion in this project is the Aquaculture and Fisheries Science Department at Bunda College of Agriculture, Malawi which is responsible for training students in aquaculture and fisheries science at the undergraduate level.Improvement in the teaching of fish processing and quality management in the Department of Aquaculture and Fisheries Science was identified as the major gap requiring action. The current teaching syllabus was thus analysed to identify weak areas. In conclusion, the project developed (as the major output) a teaching handbook for the Department of Aquaculture and Fisheries Science.

Studies on the post-mortem changes in shrimp and prawn during ice storage. Pt. 1. Organoleptic and physical changes

The organoleptic characteristics such as appearance, textural condition, colour and odour indicated that the M. rosenbergii stored in ice for 5-6 days was acceptable for processing in the industry while P. monodon under similar ice storage condition was acceptable for 8-9 days. In both species, samples stored in headless condition in ice had longer shelf life than that of stored in head-on condition. Physical changes were evaluated by determining expressible moisture and breaking strength of sample of muscles. The expressible moisture increased continuously in both samples with the lapse of storage period. The expressible moisture increased up to around 44% in 4-5 days of ice stored M. rosenbergii muscle while it was around 40% in 8-9 days ice stored P. monodon. At the end of 9 days of ice storage, the expressible moisture content in M. rosenbergii increased up to 60%, while it was up to 47% in P. monodon after 11 days of ice storage. The breaking strength declined from 0. 78 kg/cm² to 0.53 kg/cm² in tiger shrimp after 8 days of ice storage, while in case of immediately killed prawn, the breaking strength of muscle was 0.8 kg/cm² which declined to 0.43 to 0.35 kg/cm².

RNA processing and Arabidopsis flowering time control.

Plants control their flowering time in order to ensure that they reproduce under favourable conditions. The components involved in this complex process have been identified using a molecular genetic approach in Arabidopsis and classified into genetically separable pathways. The autonomous pathway controls the level of mRNA encoding a floral repressor, FLC, and comprises three RNA-binding proteins, FCA, FPA and FLK. FCA interacts with the 3'-end RNA-processing factor FY to autoregulate its own expression post-transcriptionally and to control FLC. Other components of the autonomous pathway, FVE and FLD, regulate FLC epigenetically. This combination of epigenetic and post-transcriptional control gives precision to the control of FLC expression and flowering time.

Dissecting Arabidopsis thaliana DICER function in small RNA processing, gene silencing and DNA methylation patterning.

Small RNAs have several important biological functions. MicroRNAs (miRNAs) and trans-acting small interfering RNAs (tasiRNAs) regulate mRNA stability and translation, and siRNAs cause post-transcriptional gene silencing of transposons, viruses and transgenes and are important in both the establishment and maintenance of cytosine DNA methylation. Here, we study the role of the four Arabidopsis thaliana DICER-LIKE genes (DCL1-DCL4) in these processes. Sequencing of small RNAs from a dcl2 dcl3 dcl4 triple mutant showed markedly reduced tasiRNA and siRNA production and indicated that DCL1, in addition to its role as the major enzyme for processing miRNAs, has a previously unknown role in the production of small RNAs from endogenous inverted repeats. DCL2, DCL3 and DCL4 showed functional redundancy in siRNA and tasiRNA production and in the establishment and maintenance of DNA methylation. Our studies also suggest that asymmetric DNA methylation can be maintained by pathways that do not require siRNAs.

Village level processing techniques

The agar-bearing seaweeds Gracilaria and Gelidium grow abundantly in the Asia-Pacific region. Production and post-harvest techniques and methods for processing Gracilaria to produce agar suitable for local market is necessary to increase the meager income of coastal dwellers. A flow diagram of a village level agar production is provided. A guide is also given for the quality of dried seaweeds, which are divided into 3 classes.

Rapid entropy-based detection and properties measurement of concrete spalling with machine vision for post-earthquake safety assessments

The current procedures in post-earthquake safety and structural assessment are performed manually by a skilled triage team of structural engineers/certified inspectors. These procedures, and particularly the physical measurement of the damage properties, are time-consuming and qualitative in nature. This paper proposes a novel method that automatically detects spalled regions on the surface of reinforced concrete columns and measures their properties in image data. Spalling has been accepted as an important indicator of significant damage to structural elements during an earthquake. According to this method, the region of spalling is first isolated by way of a local entropy-based thresholding algorithm. Following this, the exposure of longitudinal reinforcement (depth of spalling into the column) and length of spalling along the column are measured using a novel global adaptive thresholding algorithm in conjunction with image processing methods in template matching and morphological operations. The method was tested on a database of damaged RC column images collected after the 2010 Haiti earthquake, and comparison of the results with manual measurements indicate the validity of the method.

Enhancement of conductivity and transmittance of ZnO films by post hydrogen plasma treatment

We studied the effects of hydrogen plasma treatment on the electrical and optical properties of ZnO films deposited by radio frequency magnetron sputtering. It is found that the ZnO H film is highly transparent with the average transmittance of 92% in the visible range. Both carrier concentration and mobility are increased after hydrogen plasma treatment, correspondingly, the resistivity of the ZnO H films achieves the order of 10(-3) cm. We suggest that the incorporated hydrogen not only passivates most of the defects and/or acceptors present, but also introduces shallow donor states such as the V-O-H complex and the interstitial hydrogen H-i. Moreover, the annealing data indicate that H-i is unstable in ZnO, while the V-O-H complex remains stable on the whole at 400 degrees C, and the latter diffuses out when the annealing temperature increases to 500 degrees C. These results make ZnO H more attractive for future applications as transparent conducting electrodes.

Structural properties of polycrystalline silicon films formed by pulsed rapid thermal processing

A novel pulsed rapid thermal processing (PRTP) method has been used for realizing solid-phese crystallization of amorphous silicon films prepared by plasma-enhanced chemical vapour deposit ion. The microstructure and surface morphology of the crystallized films were investigated using x-ray diffraction and atomic Force microscopy. The results indicate that PRTP is a suitable post-crystallization technique for fabricating large-area polycrystalline silicon films with good structural quality, such as large grain size, small lattice microstrain and smooth surface morphology on low-cost glass substrates.

Structural properties of polycrystalline silicon films formed by pulsed rapid thermal processing

A novel pulsed rapid thermal processing (PRTP) method has been used for realizing the solid-phase crystallization of amorphous silicon films prepared by PECVD. The microstructure and surface morphology of the crystallized films are investigated by X-ray diffraction (XRD) and atomic force microscopy (AFM). The results indicate that this PRTP is a suitable post-crystallization technique for fabricating large-area polycrystalline silicon films with good structural qualities such as large grain size, small lattice microstain and smooth surface morphology on low-cost substrate.

Functional neuroimaging of emotionally intense autobiographical memories in post-traumatic stress disorder.

Post-traumatic stress disorder (PTSD) affects regions that support autobiographical memory (AM) retrieval, such as the hippocampus, amygdala and ventral medial prefrontal cortex (PFC). However, it is not well understood how PTSD may impact the neural mechanisms of memory retrieval for the personal past. We used a generic cue method combined with parametric modulation analysis and functional MRI (fMRI) to investigate the neural mechanisms affected by PTSD symptoms during the retrieval of a large sample of emotionally intense AMs. There were three main results. First, the PTSD group showed greater recruitment of the amygdala/hippocampus during the construction of negative versus positive emotionally intense AMs, when compared to controls. Second, across both the construction and elaboration phases of retrieval the PTSD group showed greater recruitment of the ventral medial PFC for negatively intense memories, but less recruitment for positively intense memories. Third, the PTSD group showed greater functional coupling between the ventral medial PFC and the amygdala for negatively intense memories, but less coupling for positively intense memories. In sum, the fMRI data suggest that there was greater recruitment and coupling of emotional brain regions during the retrieval of negatively intense AMs in the PTSD group when compared to controls.

Cell-specific processing of chromogranin A in endocrine cells of the rat stomach

The rat stomach is rich in endocrine cells. The acid-producing (oxyntic) mucosa contains ECL cells, A-like cells, and somatostatin (D) cells, and the antrum harbours gastrin (G) cells, enterochromaffin (EC) cells and D cells. Although chromogranin A (CgA) occurs in all these cells, its processing appears to differ from one cell type to another. Eleven antisera generated to different regions of rat CgA, two antisera generated to a human (h) CgA sequences, and one to a bovine Ib) CgA sequence, respectively, were employed together with antisera directed towards cell-specific markers such as gastrin (G cells), serotonin (EC cells), histidine decarboxylsae (ECL cells) and somatostatin (D cells) to characterize the expression of CgA and CgA-derived peptides in the various endocrine cell populations of the rat stomach. In the oxyntic mucosa, antisera raised against CgA(291-319) and CGA(316-321) immunostained D cells exclusively, whereas antisera raised against bCgA(82-91) and CgA(121-128) immunostained A-like cells and D cells. Antisera raised against CgA(318-349) and CgA(437-448) immunostained ECL cells and A-like cells, but not D cells. In the antrum, antisera against CgA(291-319) immunostained D cells, and antisera against CgA(351-356) immunostained G cells. Our observations suggest that each individual endocrine cell type in the rat stomach generates a unique mixture of CgA-derived peptides, probably reflecting cell-specific differences in the post-translational processing of CgA and its peptide products. A panel of antisera that recognize specific domains of CgA may help to identify individual endocrine cell populations.

Post-slaughter changes in ATP metabolites, Reducing and phosphorylated sugars in chicken meat

The formation of ATP breakdown products in chicken M. pectoralis major post-slaughter is reported. The concentrations of metabolites were followed in chicken breast throughout the carcass processing post-slaughter and during chilled storage. The concentration of glucose remains similar throughout the period whilst that of glucose-6-phosphate decreases linearly. Glucose and glucose-6-phosphate concentrations were inversely related to the pHu of the breast meat throughout chilled storage. Rapid post-mortem glycolysis and high pHu values suggest the occurrence of stress at and pre-slaughter. Whilst ATP, ADP and AMP were rapidly broken down, the concentration of IMP rose rapidly and remained high. Concentrations of inosine, ribose and hypoxanthine increased gradually post-slaughter but an initial increase in ribose phosphate was not sustained. Most of the potential ribose present in chicken meat, believed to be important for flavor formation, remains bound in the form of inosine and IMP. There is evidence that additional breakdown pathways for ribose and ribose-5-phosphate may deplete the concentrations of these precursors.