Feasibility Study of Reclaimed Asphalt Pavements (RAP) as Recycled Aggregates Used in Rigid Pavement Construction.

Reclaimed Asphalt Pavement (RAP) as recycled aggregates is a relatively new construction process of rigid pavements due to the scarcity and degradation of natural aggregates. This study aims at the sequential characterization of RAP aggregate to obtain optimized proportions for strength. For this purpose, RAP aggregates were used for the replacement of natural aggregates (NA) in the concrete mix which was achieved by varying from 0-100%. Furthermore, zirconia silica fume (ZSF) was used as a partial replacement of the cement in the concrete mix, replacing Ordinary Portland Cement (OPC). Experimental studies have shown that the incorporation of washed RAP (WRAP) slightly reduces the compressive strength of concrete by 2.7-37.35% as compared to the reference control concrete mix. Although the 7-days, 28-days and 56-day compressive strength of WRAP recycled aggregate-based concrete is slightly better than the 7-days, 28-days and 56-day compressive strength of dirty RAP (DRAP) recycled aggregate-based concrete. A similar trend was observed in the flexural strength and split tensile strength of WRAP recycled aggregate-based. Overall, the results show that 40% WRAP recycled aggregates with 10% ZSF as a replacement for cement outperform DRAP aggregates in concrete mixes. According to the ANOVA results, the combination of ZSF and WRAP aggregates met the cement concrete pavement strength standard, thereby contributing to sustainable development. Reclaimed Asphalt Concrete Pavements (RACP) are now seen as a potential and long-term answer to the present environmental and economic crisis.

Foot-and-mouth disease virus transmission dynamics and persistence in a herd of vaccinated dairy cattle in India.

Foot-and-mouth disease (FMD) is an important transboundary disease with substantial economic impacts. Although between-herd transmission of the disease has been well studied, studies focusing on within-herd transmission using farm-level outbreak data are rare. The aim of this study was to estimate parameters associated with within-herd transmission, host physiological factors and FMD virus (FMDV) persistence using data collected from an outbreak that occurred at a large, organized dairy farm in India. Of 1,836 regularly vaccinated, adult dairy cattle, 222 had clinical signs of FMD over a 39-day period. Assuming homogenous mixing, a frequency-dependent compartmental model of disease transmission was built. The transmission coefficient and basic reproductive number were estimated to be between 16.2-18.4 and 67-88, respectively. Non-pregnant animals were more likely to manifest clinical signs of FMD as compared to pregnant cattle. Based on oropharyngeal fluid (probang) sampling and FMDV-specific RT-PCR, four of 36 longitudinally sampled animals (14%) were persistently infected carriers 10.5 months post-outbreak. There was no statistical difference between subclinical and clinically infected animals in the duration of the carrier state. However, prevalence of NSP-ELISA antibodies differed significantly between subclinical and clinically infected animals 12 months after the outbreak with 83% seroprevalence amongst clinically infected cattle compared to 69% of subclinical animals. This study further elucidates within-herd FMD transmission dynamics during the acute-phase and characterizes duration of FMDV persistence and seroprevalence of FMD under natural conditions in an endemic setting.

Quantitative characteristics of the foot-and-mouth disease carrier state under natural conditions in India.

The goal of this study was to characterize the properties and duration of the foot-and-mouth disease (FMD) carrier state and associated serological responses subsequent to vaccination and naturally occurring infection at two farms in northern India. Despite previous vaccination of cattle in these herds, clinical signs of FMD occurred in October 2013 within a subset of animals at the farms containing juvenile-yearling heifers and steers (Farm A) and adult dairy cattle (Farm B). Subsequent to the outbreak, FMD virus (FMDV) asymptomatic carriers were identified in both herds by seroreactivity to FMDV non-structural proteins and detection of FMDV genomic RNA in oropharyngeal fluid. Carriers' seroreactivity and FMDV genome detection status were subsequently monitored monthly for 23 months. The mean extinction time of the carrier state was 13.1 ± 0.2 months, with extinction having occurred significantly faster amongst adult dairy cattle at Farm B compared to younger animals at Farm A. The rate of decrease in the proportion of carrier animals was calculated to be 0.07 per month. Seroprevalence against FMDV non-structural proteins decreased over the course of the study period, but was found to increase transiently following repeated vaccinations. These data provide novel insights into viral and host factors associated with the FMDV carrier state under natural conditions. The findings reported herein may be relevant to field veterinarians and governmental regulatory entities engaged in FMD response and control measures.

Virus load in pigs affected with different clinical forms of classical swine fever.

Classical swine fever (CSF) is an endemic disease in India, but the real magnitude of the problem is not known as only outbreaks of acute CSF are reported and many cases of chronic and clinically inapparent forms of the disease, which manifest a confusing clinical picture, remain undiagnosed. The real status of classical swine fever virus (CSFV) infection can only be known by testing pigs with highly specific and sensitive diagnostic assays. To obtain the baseline prevalence of CSFV infection among pigs in an endemic region where no vaccination was being performed, a real-time PCR assay was used to detect viral genetic material in tissue samples collected from a slaughterhouse in the northern state of Uttar Pradesh in India. In total, 1120 slaughtered pigs were examined for the presence of CSF suggestive pathological lesions and tissues from suspected cases were tested for the presence of CSFV antigen and nucleic acids by indirect immuno-peroxidase test and real-time PCR, respectively. Based on the detection of viral genetic material in the tonsils, the prevalence of CSFV infection among slaughtered pigs was found to be 7.67%. Pigs detected positive for viral genome by quantitative real-time PCR assay when categorized into different forms of CSF, depending upon the pathological lesions observed, the viral load in the tonsils of some of the pigs with chronic or clinically inapparent form of the disease was similar to that detected in pigs with acute CSF. The results of the study suggested that the risk posed by pigs with chronic disease or those infected but showing no clinical disease may be relatively higher as they can transmit the virus to new susceptible hosts over a longer period of time.

Serological evidence of foot-and-mouth disease virus infection in randomly surveyed goat population of Orissa, India.

India is endemic for foot-and-mouth disease (FMD), and goats constitute the second largest susceptible population of domestic livestock. FMD surveillance and control strategies in the country largely ignore small ruminants, known to be critical in the epidemiology of the disease. Here, serological investigations were carried out to generate estimates of antibody prevalence in goats of Orissa state to both non-structural (NSP-Ab) and structural proteins (SP-Ab) of FMD. The apparent overall NSP-Ab and SP-Ab seroprevalences were 38% and 20.7%, respectively, which signifies a very high level of FMD virus circulation in the goat population despite the lack of clinical signs in this species. The apparent prevalence of NSP-Ab and SP-Ab was positively correlated in the sampling areas. Interestingly, the values found for NSP-Ab prevalence were almost consistently higher than those found for SP-Ab prevalence. This could have been attributable to either issues related to sensitivity and specificity of the test systems employed or differences in the post-infection kinetics of NSP- and SP-Ab. The pattern that emerged from SP-Ab analysis indicated goats being infected with all three prevalent serotypes (O, A and Asia 1) and reinforces the concept that non-vaccinated goats can be exploited as tracer animals for detecting serotypes involved in outbreaks. The results underscore the requirement to bring caprine species under comprehensive surveillance and vaccination campaigns to check silent amplification, excretion and transmission of the virus.

The mechanism of nucleocytoplasmic transport through the nuclear pore complex.

The nuclear pore complex (NPC) mediates all transport between the nucleus and cytoplasm. Passage through the NPC is highly selective, yet the same channel must allow rapid specific transport of a wide range of cargoes. This chapter focuses mainly on the phenylalanine-glycine (FG) nucleoporins (nups), proteins carrying natively unfolded regions that are thought to form the selectively permeable barrier within the NPC. The physical properties of the FG nup barrier remain unclear. The high selectivity and rapidity of transport observed in vivo may be explained, in part, by competition for binding and space between transport factors and nontransported proteins. Future studies of FG nups will therefore also examine their interactions between FG nups and other proteins in their surroundings.

Reducing primary and secondary impact loads on the pelvis during side impact.

OBJECTIVE: The objective of the study was to determine which vehicle factors are significantly related to pelvic injury in side impact collisions. Identification of relevant parameters could aid in the reduction of these injuries. METHOD: Side impact crashes from the CIREN database were separated into those in which the occupant sustained a pelvic fracture and those in which no pelvic fracture occurred, although all occupants had serious injuries. A multibody MADYMO model was created of a USDOT SINCAP (U.S. Department of Transportation Side Impact New Car Assessment Program) test of a vehicle with a large center console. RESULTS: From a study of 113 side impact crashes in the ciren database, nearside occupants with pelvic fractures (n = 78) had (i) more door intrusion (mean, 37 vs. 32 cm, p = 0.02) than those who had serious injuries, but not pelvic fractures (ii) a greater likelihood that the lower border of the door intruded more than the upper part (40% vs. 18%, p < 0.025); and (iii) a greater likelihood that their vehicle had a center console (47 vs. 17%, p < 0.005). Other parameters such as occupant age, weight, gender, vehicle weight, and struck vehicle speed change were not significantly different. MADYMO modeling showed that with a center console, an initial positive pelvic acceleration occurred at about 30 msec, followed at about 45 msec by a second acceleration peak in the opposite direction. Reducing console stiffness reduced the second acceleration but not the initial peak. Allowing the seat to translate laterally when contacted by the door reduced the initial pelvic acceleration by 50% and eliminated the second acceleration peak. CONCLUSIONS: Redesigning the center console using less stiff materials and allowing some lateral translation of the seat could aid in reducing pelvic injuries in side impact collisions.

Composition and functional characterization of yeast 66S ribosome assembly intermediates.

The pathway and complete collection of factors that orchestrate ribosome assembly are not clear. To address these problems, we affinity purified yeast preribosomal particles containing the nucleolar protein Nop7p and developed means to separate their components. Nop7p is associated primarily with 66S preribosomes containing either 27SB or 25.5S plus 7S pre-rRNAs. Copurifying proteins identified by mass spectrometry include ribosomal proteins, nonribosomal proteins previously implicated in 60S ribosome biogenesis, and proteins not known to be involved in ribosome production. Analysis of strains mutant for eight of these proteins not previously implicated in ribosome biogenesis showed that they do participate in this pathway. These results demonstrate that proteomic approaches in concert with genetic tools provide powerful means to purify and characterize ribosome assembly intermediates.

Isolation and characterization of subnuclear compartments from Trypanosoma brucei. Identification of a major repetitive nuclear lamina component.

Protozoan parasites of the order Kinetoplastida are responsible for a significant proportion of global morbidity and economic hardship. These organisms also represent extremely distal points within the Eukarya, and one such organism, Trypanosoma brucei, has emerged as a major system for the study of evolutionary cell biology. Significant technical challenges have hampered the full exploitation of this organism, but advances in genomics and proteomics provide a novel approach to acquiring rapid functional data. However, the vast evolutionary distance between trypanosomes and the higher eukaryotes presents significant problems with functional assignment based on sequence similarity, and frequently homologues cannot be identified with sufficient confidence to be informative. Direct identification of proteins in isolated organelles has the potential of providing robust functional insight and is a powerful approach for initial assignment. We have selected the nucleus of T. brucei as a first target for protozoan organellar proteomics. Our purification methodology was able to reliably provide both nuclear and subnuclear fractions. Analysis by gel electrophoresis, electron microscopy, and immunoblotting against trypanosome subcellular markers indicated that the preparations are of high yield and purity, maintain native morphology, and are well resolved from other organelles. Minor developmental differences were observed in the nuclear proteome for the bloodstream and procyclic stages, whereas significant morphological alterations were visible. We demonstrate by direct sequencing that the NUP-1 nuclear envelope antigen is a coiled coil protein, containing approximately 20 near-perfect copies of a 144-amino acid sequence. Immunoelectron microscopy localized NUP-1 to the inner face of the nuclear envelope, suggesting that it is a major filamentous component of the trypanosome nuclear lamina.

Study of oat globulin conformation by Fourier transform infrared spectroscopy.

The conformation of oat globulin dispersions (10% in D2O) under the influence of pH, chaotropic salts, protein structure perturbants, and heating conditions was studied by Fourier transform infrared (FTIR) spectroscopy. The FTIR spectrum of oat globulin showed major bands from 1670 to 1634 cm(-1), corresponding to the four major types of secondary structures, that is, beta-turns, beta-sheets, alpha-helices, and random coils. At extreme acidic and alkaline pH conditions, there were changes in intensity in the bands attributed to beta-sheet structures (1626, 1634, and 1682 cm(-1)), and shifts of the bands to higher or lower wavenumbers, indicating changes in conformation. In the presence of some chaotropic salts, the 1626 and 1634 cm(-1) bands were shifted upward, with a marked decrease in the intensity of the 1634 cm(-1) peak. The addition of several protein structure perturbants led to a slight shift in the alpha-helix/random coil bands and a marked reduction in the beta-sheet peaks, suggesting protein unfolding. Heating under aggregating conditions led to slight shifts in all of the major bands and progressive changes in the intensity of the alpha-helix, beta-sheet, and beta-turn peaks, suggesting protein denaturation. This was accompanied by marked increases in intensity of the two intermolecular beta-sheet bands (1682 and 1624-1626 cm(-1)) associated with the formation of aggregated strands. The IR spectra of soluble and insoluble aggregates showed a redistribution of native and extensively denatured proteins in the two fractions.

Nup2p dynamically associates with the distal regions of the yeast nuclear pore complex.

Nucleocytoplasmic transport is mediated by the interplay between soluble transport factors and nucleoporins resident within the nuclear pore complex (NPC). Understanding this process demands knowledge of components of both the soluble and stationary phases and the interface between them. Here, we provide evidence that Nup2p, previously considered to be a typical yeast nucleoporin that binds import- and export-bound karyopherins, dynamically associates with the NPC in a Ran-facilitated manner. When bound to the NPC, Nup2p associates with regions corresponding to the nuclear basket and cytoplasmic fibrils. On the nucleoplasmic face, where the Ran--GTP levels are predicted to be high, Nup2p binds to Nup60p. Deletion of NUP60 renders Nup2p nucleoplasmic and compromises Nup2p-mediated recycling of Kap60p/Srp1p. Depletion of Ran--GTP by metabolic poisoning, disruption of the Ran cycle, or in vitro by cell lysis, results in a shift of Nup2p from the nucleoplasm to the cytoplasmic face of the NPC. This mobility of Nup2p was also detected using heterokaryons where, unlike nucleoporins, Nup2p was observed to move from one nucleus to the other. Together, our data support a model in which Nup2p movement facilitates the transition between the import and export phases of nucleocytoplasmic transport.

Traditional Phytotherapy for Diarroeal Diseases in Dhenkanal district of Orissa, India.

Medico- etnobotanical exploration carried out in Dhenkanal district of Orissa during 1996-98 reveal that, people use 21 plant species belonging o 20 genera and 17 families on 10 different combinations for the treatment of diarrehoeal diseases. The method of preparation of medicine and details of application care recorded.

Raman spectroscopic study of oat globulin conformation.

Analysis of Raman spectra of oat globulin showed that extreme pH values caused an increase in the amide and C-H stretching band intensity, indicating changes in the secondary structures of the protein due to denaturation. Similar changes were observed when oat globulin was treated with chaotropic salts and several protein perturbants. Sodium dodecyl sulfate, beta-mercaptoethanol, and ethylene glycol also caused a shift in the amide III' band, suggesting a transition from beta-sheet to a random coil conformation. Heating at temperatures near the denaturation temperature of oat globulin led to increases in the amide and C-H band intensity, indicating unfolding of the protein. The data indicate that FT-Raman spectroscopy is suitable for studying the secondary structure of plant proteins such as oat globulin.

The yeast nuclear pore complex: composition, architecture, and transport mechanism.

An understanding of how the nuclear pore complex (NPC) mediates nucleocytoplasmic exchange requires a comprehensive inventory of the molecular components of the NPC and a knowledge of how each component contributes to the overall structure of this large molecular translocation machine. Therefore, we have taken a comprehensive approach to classify all components of the yeast NPC (nucleoporins). This involved identifying all the proteins present in a highly enriched NPC fraction, determining which of these proteins were nucleoporins, and localizing each nucleoporin within the NPC. Using these data, we present a map of the molecular architecture of the yeast NPC and provide evidence for a Brownian affinity gating mechanism for nucleocytoplasmic transport.

Pore relations: nuclear pore complexes and nucleocytoplasmic exchange.

NPCs are the sole sites of exchange between the nucleus and cytoplasm. A large family of transport factors carry cargo between the nucleus and cytoplasm through the NPC. The NPC is a huge symmetric octagonal structure comprised of dozens of NUPs. NUPs make many contacts with surrounding structures, including the NE, the cytoplasm and nuclear interior. A subset of NUPs contain repeated peptide motifs that serve as docking sites for transport factors. The directionality of transport is determined by the transport factor, and its interactions with the small GTPase Ran and NUPs. Very little is known about how the NPC mediates transport, NPC assembly and the NPC's role in regulating transport, but these areas of research are beginning to emerge.

The lysine and methionine rich basic subunit of buckwheat grain legumin: some results of a structural study.

The 26 kD basic subunit of 280 kD buckwheat grain legumin has been partially characterized by measurement of its fluorescence and CD spectra. The protein has 22% alpha-helix, 36% beta-sheet, 12% beta-turn and 30% random coil secondary structure. In comparison with the basic subunits of other legumin-type proteins, the buckwheat legumin subunit has a high content of lysine and methionine. The protein also has higher ratios of lysine to arginine and methionine to arginine.

Proteins connecting the nuclear pore complex with the nuclear interior.

While much has been learned in recent years about the movement of soluble transport factors across the nuclear pore complex (NPC), comparatively little is known about intranuclear trafficking. We isolated the previously identified Saccharomyces protein Mlp1p (myosin-like protein) by an assay designed to find nuclear envelope (NE) associated proteins that are not nucleoporins. We localized both Mlp1p and a closely related protein that we termed Mlp2p to filamentous structures stretching from the nucleoplasmic face of the NE into the nucleoplasm, similar to the homologous vertebrate and Drosophila Tpr proteins. Mlp1p can be imported into the nucleus by virtue of a nuclear localization sequence (NLS) within its COOH-terminal domain. Overexpression experiments indicate that Mlp1p can form large structures within the nucleus which exclude chromatin but appear highly permeable to proteins. Remarkably, cells harboring a double deletion of MLP1 and MLP2 were viable, although they showed a slower net rate of active nuclear import and faster passive efflux of a reporter protein. Our data indicate that the Tpr homologues are not merely NPC-associated proteins but that they can be part of NPC-independent, peripheral intranuclear structures. In addition, we suggest that the Tpr filaments could provide chromatin-free conduits or tracks to guide the efficient translocation of macromolecules between the nucleoplasm and the NPC.