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Sample records for anti-white spot syndrome

  1. Anti-white spot syndrome virus activity of Ceriops tagal aqueous extract in giant tiger shrimp Penaeus monodon.

    PubMed

    Sudheer, N S; Philip, Rosamma; Bright Singh, I S

    2012-09-01

    White spot syndrome virus (WSSV), the most contagious pathogen of cultured shrimp, causes mass mortality, leading to huge economic loss to the shrimp industry. The lack of effective therapeutic or prophylactic measures has aggravated the situation, necessitating the development of antiviral agents. With this objective, the antiviral activity in the aqueous extract of a mangrove plant Ceriops tagal in Penaeus monodon was evaluated. The Ceriops tagal aqueous extract (CTAE) was non-toxic to shrimps at 50 mg/ml when injected intramuscularly at a dosage of 10 μL/animal (0.5 mg/animal) and showed a protective effect against WSSV at 30 mg/ml when mixed with WSSV suspension at a 1:1 ratio. When the extract was administered along with the diet and the animals were challenged orally, there was a dose-dependent increase in survival, culminating in 100 % survival at a concentration of 500 mg/kg body weight/day. Neither hypertrophied nuclei nor the viral envelope protein VP28 could be demonstrated in surviving shrimps using histology and indirect immunofluorescence histochemistry (IIFH), respectively. To elucidate the mode of action, the temporal expression of WSSV genes and shrimp immune genes, including antimicrobial peptides, was attempted. None of the viral genes were found to be expressed in shrimps that were fed with the extract and challenged or in those that were administered CTAE-exposed WSSV. The overall results suggest that the aqueous extract from C. tagal can protect P. monodon from white spot syndrome virus infection.

  2. Screening, isolation and optimization of anti–white spot syndrome virus drug derived from terrestrial plants

    PubMed Central

    Ghosh, Upasana; Chakraborty, Somnath; Balasubramanian, Thangavel; Das, Punyabrata

    2014-01-01

    Objective To screen, isolate and optimize anti-white spot syndrome virus (WSSV) drug derived from various terrestrial plants and to evaluate the efficacy of the same in host–pathogen interaction model. Methods Thirty plants were subjected to Soxhlet extraction using water, ethanol, methanol and hexane as solvents. The 120 plant isolates thus obtained were screened for their in vivo anti–WSSV property in Litopenaeus vannamei. The best anti–WSSV plant isolate, TP22C was isolated and further analyzed. The drug was optimized at various concentrations. Viral and immune genes were analysed using reverse transcriptase PCR to confirm the potency of the drug. Results Seven plant isolates exhibited significant survivability in host. The drug TP22C thus formulated showed 86% survivability in host. The surviving shrimps were nested PCR negative at the end of the 15 d experimentation. The lowest concentration of TP22C required intramuscularly for virucidal property was 10 mg/mL. The oral dosage of 750 mg/kg body weight/day survived at the rate of 86%. Neither VP28 nor ie 1 was expressed in the test samples at 42nd hour and 84th hour post viral infection. Conclusions The drug TP22C derived from Momordica charantia is a potent anti-white spot syndrome virus drug. PMID:25183066

  3. Screening, isolation and optimization of anti–white spot syndrome virus drug derived from marine plants

    PubMed Central

    Chakraborty, Somnath; Ghosh, Upasana; Balasubramanian, Thangavel; Das, Punyabrata

    2014-01-01

    Objective To screen, isolate and optimize anti-white spot syndrome virus (WSSV) drug derived from various marine floral ecosystems and to evaluate the efficacy of the same in host–pathogen interaction model. Methods Thirty species of marine plants were subjected to Soxhlet extraction using water, ethanol, methanol and hexane as solvents. The 120 plant isolates thus obtained were screened for their in vivo anti-WSSV property in Litopenaeus vannamei. By means of chemical processes, the purified anti-WSSV plant isolate, MP07X was derived. The drug was optimized at various concentrations. Viral and immune genes were analysed using reverse transcriptase PCR to confirm the potency of the drug. Results Nine plant isolates exhibited significant survivability in host. The drug MP07X thus formulated showing 85% survivability in host. The surviving shrimps were nested PCR negative at the end of the 15 d experimentation. The lowest concentration of MP07X required intramuscularly for virucidal property was 10 mg/mL. The oral dosage of 1 000 mg/kg body weight/day survived at the rate of 85%. Neither VP28 nor ie 1 was expressed in the test samples at 42nd hour and 84th hour post viral infection. Conclusions The drug MP07X derived from Rhizophora mucronata is a potent anti-WSSV drug. PMID:25183065

  4. LEOPARD syndrome: what are café noir spots?

    PubMed

    Rodríguez-Bujaldón, Alfonso; Vazquez-Bayo, Carmen; Jimenez-Puya, Rafael; Galan-Gutierrez, Manuel; Moreno-Gimenez, José; Rodriguez-Garcia, Alfonso; Tercedor, Jesus; Velez-Garcia, Antonio

    2008-01-01

    Lentigines, electrocardiographic abnormalities, ocular hypertelorism, pulmonary stenosis, abnormalities of genitalia, retarded growth, and deafness syndrome (multiple lentigines syndrome) is most often characterized by multiple lentigines and cardiac conduction defects. Café noir spot is a term proposed, by analogy to café au lait spots, for the larger and darkly pigmented patches that are frequently observed in patients with this syndrome. Although presumed by some authors to represent lentigines, the histologic features of café noir spots have not been well documented in the literature. Only two previous cases have been reported in which a biopsy of the café noir spots than melanocytic nevi. We describe the histologic characteristics of seven café noir spots in six patients with lentigines, electrocardiographic abnormalities, ocular hypertelorism, pulmonary stenosis, abnormalities of genitalia, retarded growth, and deafness syndrome. Three lesions represented melanocytic nevi (one with dysplastic features), and four were compatible with lentigo simplex. These findings help our understanding of the histologic spectrum of pigmented lesions in lentigines, electrocardiographic abnormalities, ocular hypertelorism, pulmonary stenosis, abnormalities of genitalia, retarded growth, and deafness syndrome.

  5. Does Dark-Spot Syndrome Experimentally Transmit among Caribbean Corals?

    PubMed Central

    Randall, Carly J.; Jordán-Garza, Adán G.; Muller, Erinn M.; van Woesik, Robert

    2016-01-01

    Over the last half-century, coral diseases have contributed to the rapid decline of coral populations throughout the Caribbean region. Some coral diseases appear to be potentially infectious, yet little is known about their modes of transmission. This study experimentally tested whether dark-spot syndrome on Siderastrea siderea was directly or indirectly transmissible to neighboring coral colonies. We also tested whether open wounds were necessary to facilitate disease transmission. At the completion of the experiments, we sampled bacterial communities on diseased, exposed, and healthy coral colonies to determine whether bacterial pathogens had transmitted to the susceptible colonies. We saw no evidence of either direct or waterborne transmission of dark-spot syndrome, and corals that received lesions by direct contact with diseased tissue, healed and showed no signs of infection. There were no significant differences among bacterial communities on healthy, exposed, and diseased colonies, although nine individual ribotypes were significantly higher in diseased corals compared with healthy and exposed corals, indicating a lack of transmission. Although our experiments do not fully refute the possibility that dark-spot syndrome is infectious and transmissible, our results suggest that in situ macroscopic signs of dark-spot syndrome are not always contagious. PMID:26788918

  6. Does Dark-Spot Syndrome Experimentally Transmit among Caribbean Corals?

    PubMed

    Randall, Carly J; Jordán-Garza, Adán G; Muller, Erinn M; van Woesik, Robert

    2016-01-01

    Over the last half-century, coral diseases have contributed to the rapid decline of coral populations throughout the Caribbean region. Some coral diseases appear to be potentially infectious, yet little is known about their modes of transmission. This study experimentally tested whether dark-spot syndrome on Siderastrea siderea was directly or indirectly transmissible to neighboring coral colonies. We also tested whether open wounds were necessary to facilitate disease transmission. At the completion of the experiments, we sampled bacterial communities on diseased, exposed, and healthy coral colonies to determine whether bacterial pathogens had transmitted to the susceptible colonies. We saw no evidence of either direct or waterborne transmission of dark-spot syndrome, and corals that received lesions by direct contact with diseased tissue, healed and showed no signs of infection. There were no significant differences among bacterial communities on healthy, exposed, and diseased colonies, although nine individual ribotypes were significantly higher in diseased corals compared with healthy and exposed corals, indicating a lack of transmission. Although our experiments do not fully refute the possibility that dark-spot syndrome is infectious and transmissible, our results suggest that in situ macroscopic signs of dark-spot syndrome are not always contagious.

  7. White spot syndrome virus inactivation study by using gamma irradiation

    NASA Astrophysics Data System (ADS)

    Heidareh, Marzieh; Sedeh, Farahnaz Motamedi; Soltani, Mehdi; Rajabifar, Saeed; Afsharnasab, Mohammad; Dashtiannasab, Aghil

    2014-09-01

    The present study was conducted to investigate the effect of gamma irradiation on white spot syndrome virus (WSSV). White spot syndrome virus is a pathogen of major economic importance in cultured penaeid shrimp industries. White spot disease can cause mortalities reaching 100% within 3-10 days of gross signs appearing. During the period of culture, immunostimulant agents and vaccines may provide potential methods to protect shrimps from opportunistic and pathogenic microrganisms. In this study, firstly, WSSV was isolated from infected shrimp and then multiplied in crayfish. WSSV was purified from the infected crayfish haemolymph by sucrose gradient and confirmed by transmission electron microscopy. In vivo virus titration was performed in shrimp, Penaeus semisulcatus. The LD50 of live virus stock was calculated 10 5.4/mL. Shrimp post-larvae (1-2 g) were treated with gamma-irradiated (different doses) WSSV (100 to 10-4 dilutions) for a period of 10 days. The dose/survival curve for irradiated and un-irradiated WSSV was drawn; the optimum dose range for inactivation of WSSV and unaltered antigenicity was obtained 14-15 kGy. This preliminary information suggests that shrimp appear to benefit from treatment with gammairradiated WSSV especially at 14-15 KGy.

  8. Draft Genome Sequence of White Spot Syndrome Virus Isolated from Cultured Litopenaeus vannamei in Mexico

    PubMed Central

    Rodriguez-Anaya, Libia Zulema; Gonzalez-Galaviz, Jose Reyes; Casillas-Hernandez, Ramón; Lares-Villa, Fernando; Estrada, Karel

    2016-01-01

    The first genome sequence of a Mexican white spot syndrome virus is presented here. White spot syndrome is a shrimp pandemic virus that has devastated production in Mexico for more than 10 years. The availability of this genome will greatly aid epidemiological studies worldwide, contributing to the molecular diagnostic and disease prevention in shrimp farming. PMID:26966222

  9. Draft Genome Sequence of White Spot Syndrome Virus Isolated from Cultured Litopenaeus vannamei in Mexico.

    PubMed

    Rodriguez-Anaya, Libia Zulema; Gonzalez-Galaviz, Jose Reyes; Casillas-Hernandez, Ramón; Lares-Villa, Fernando; Estrada, Karel; Ibarra-Gamez, Jose Cuauhtemoc; Sanchez-Flores, Alejandro

    2016-03-10

    The first genome sequence of a Mexican white spot syndrome virus is presented here. White spot syndrome is a shrimp pandemic virus that has devastated production in Mexico for more than 10 years. The availability of this genome will greatly aid epidemiological studies worldwide, contributing to the molecular diagnostic and disease prevention in shrimp farming.

  10. White spot syndrome virus: an overview on an emergent concern

    PubMed Central

    Sánchez-Paz, Arturo

    2010-01-01

    Viruses are ubiquitous and extremely abundant in the marine environment. One of such marine viruses, the white spot syndrome virus (WSSV), has emerged globally as one of the most prevalent, widespread and lethal for shrimp populations. However, at present there is no treatment available to interfere with the unrestrained occurrence and spread of the disease. The recent progress in molecular biology techniques has made it possible to obtain information on the factors, mechanisms and strategies used by this virus to infect and replicate in susceptible host cells. Yet, further research is still required to fully understand the basic nature of WSSV, its exact life cycle and mode of infection. This information will expand our knowledge and may contribute to developing effective prophylactic or therapeutic measures. This review provides a state-of-the-art overview of the topic, and emphasizes the current progress and future direction for the development of WSSV control strategies. PMID:20181325

  11. White spot syndrome virus: an overview on an emergent concern.

    PubMed

    Sánchez-Paz, Arturo

    2010-01-01

    Viruses are ubiquitous and extremely abundant in the marine environment. One of such marine viruses, the white spot syndrome virus (WSSV), has emerged globally as one of the most prevalent, widespread and lethal for shrimp populations. However, at present there is no treatment available to interfere with the unrestrained occurrence and spread of the disease. The recent progress in molecular biology techniques has made it possible to obtain information on the factors, mechanisms and strategies used by this virus to infect and replicate in susceptible host cells. Yet, further research is still required to fully understand the basic nature of WSSV, its exact life cycle and mode of infection. This information will expand our knowledge and may contribute to developing effective prophylactic or therapeutic measures. This review provides a state-of-the-art overview of the topic, and emphasizes the current progress and future direction for the development of WSSV control strategies.

  12. Molecular epidemiology of white spot syndrome virus within Vietnam.

    PubMed

    Dieu, Bui Thi Minh; Marks, Hendrik; Siebenga, J Joukje; Goldbach, Rob W; Zuidema, Douwe; Duong, Tran Phuoc; Vlak, Just M

    2004-12-01

    White spot syndrome virus (WSSV), the sole member of the virus family Nimaviridae, is a large double-stranded DNA virus that infects shrimp and other crustaceans. By alignment of three completely sequenced isolates originating from Taiwan (WSSV-TW), China (WSSV-CN) and Thailand (WSSV-TH), the variable loci in the genome were mapped. The variation suggests the spread of WSSV from a common ancestor originating from either side of the Taiwan Strait to Thailand, but support for this hypothesis through analysis of geographical intermediates is sought. RFLP analysis of eight Vietnamese WSSV isolates, of which six were collected along the central coast (VN-central) and two along the south coast (VN-south), showed apparent sequence variation in the variable loci identified previously. These loci were characterized in detail by PCR amplification, cloning and sequencing. Relative to WSSV-TW, all VN-central isolates showed a approximately 8.5 kb deletion in the major variable region ORF23/24, whereas the VN-south isolates contain a deletion of approximately 11.5 or approximately 12.2 kb, compared to a approximately 1.2 or approximately 13.2 kb deletion in WSSV-CN and WSSV-TH, respectively. The minor variable region ORF14/15 showed deletions of various sizes compared with WSSV-TH for all eight VN isolates. The data suggest that the VN isolates and WSSV-TH have a common lineage, which branched off from WSSV-TW and WSSV-CN early on, and that WSSV entered Vietnam by multiple introductions. A model is presented for the spread of WSSV from either side of the Taiwan Strait into Vietnam based on the gradually increasing deletions of both 'variable regions'. The number and order of repeat units within ORF75 and ORF125 appeared to be suitable markers to study regional spread of WSSV.

  13. Suppression of shrimp melanization during white spot syndrome virus infection.

    PubMed

    Sutthangkul, Jantiwan; Amparyup, Piti; Charoensapsri, Walaiporn; Senapin, Saengchan; Phiwsaiya, Kornsunee; Tassanakajon, Anchalee

    2015-03-06

    The melanization cascade, activated by the prophenoloxidase (proPO) system, plays a key role in the production of cytotoxic intermediates, as well as melanin products for microbial sequestration in invertebrates. Here, we show that the proPO system is an important component of the Penaeus monodon shrimp immune defense toward a major viral pathogen, white spot syndrome virus (WSSV). Gene silencing of PmproPO(s) resulted in increased cumulative shrimp mortality after WSSV infection, whereas incubation of WSSV with an in vitro melanization reaction prior to injection into shrimp significantly increased the shrimp survival rate. The hemolymph phenoloxidase (PO) activity of WSSV-infected shrimp was extremely reduced at days 2 and 3 post-injection compared with uninfected shrimp but was fully restored after the addition of exogenous trypsin, suggesting that WSSV probably inhibits the activity of some proteinases in the proPO cascade. Using yeast two-hybrid screening and co-immunoprecipitation assays, the viral protein WSSV453 was found to interact with the proPO-activating enzyme 2 (PmPPAE2) of P. monodon. Gene silencing of WSSV453 showed a significant increase of PO activity in WSSV-infected shrimp, whereas co-silencing of WSSV453 and PmPPAE2 did not, suggesting that silencing of WSSV453 partially restored the PO activity via PmPPAE2 in WSSV-infected shrimp. Moreover, the activation of PO activity in shrimp plasma by PmPPAE2 was significantly decreased by preincubation with recombinant WSSV453. These results suggest that the inhibition of the shrimp proPO system by WSSV partly occurs via the PmPPAE2-inhibiting activity of WSSV453.

  14. Severe Fever with Thrombocytopenia Syndrome Complicated by Co-infection with Spotted Fever Group Rickettsiae, China.

    PubMed

    Lu, Qing-Bin; Li, Hao; Zhang, Pan-He; Cui, Ning; Yang, Zhen-Dong; Fan, Ya-Di; Cui, Xiao-Ming; Hu, Jian-Gong; Guo, Chen-Tao; Zhang, Xiao-Ai; Liu, Wei; Cao, Wu-Chun

    2016-11-01

    During 2013-2015 in central China, co-infection with spotted fever group rickettsiae was identified in 77 of 823 patients infected with severe fever with thrombocytopenia syndrome virus. Co-infection resulted in delayed recovery and increased risk for death, prompting clinical practices in the region to consider co-infection in patients with severe fever with thrombocytopenia syndrome.

  15. Severe Fever with Thrombocytopenia Syndrome Complicated by Co-infection with Spotted Fever Group Rickettsiae, China

    PubMed Central

    Lu, Qing-Bin; Li, Hao; Zhang, Pan-He; Cui, Ning; Yang, Zhen-Dong; Fan, Ya-Di; Cui, Xiao-Ming; Hu, Jian-Gong; Guo, Chen-Tao; Zhang, Xiao-Ai; Cao, Wu-Chun

    2016-01-01

    During 2013–2015 in central China, co-infection with spotted fever group rickettsiae was identified in 77 of 823 patients infected with severe fever with thrombocytopenia syndrome virus. Co-infection resulted in delayed recovery and increased risk for death, prompting clinical practices in the region to consider co-infection in patients with severe fever with thrombocytopenia syndrome. PMID:27767921

  16. Fusarium avenaceum causes burn spot disease syndrome in noble crayfish (Astacus astacus).

    PubMed

    Makkonen, J; Jussila, J; Koistinen, L; Paaver, T; Hurt, M; Kokko, H

    2013-06-01

    Burn spot disease has been causing epidemics both in the Estonian mainland and in Saaremaa Island in the threatened noble crayfish (Astacus astacus) stocks. To study the cause of the disease, we isolated several Fusarium spp. from Estonian noble crayfish (A. astacus) populations suffering from burn spot disease syndrome. We first identified fungi directly from melanised cuticle by their ITS sequences. Then we isolated Fusarium spp. from melanised spots of crayfish showing burn spot disease symptoms, such as melanisation and shell erosion, from two different crayfish populations and watercourses in Estonia. The isolates were then identified based on ITS and EF1α-gene sequences. Isolates of Fusarium spp. taken from two separate Estonian noble crayfish populations were used in infection studies. Koch postulates confirmed that the studied agent was causing burn spot disease symptoms including shell erosion in the noble crayfish, which were significantly more severe after molts. After the infection period, an identical Fusarium spp. was re-isolated from carapace lesions and was thus shown to be the disease agent causing burn spot disease syndrome and shell erosion in noble crayfish. Based on GenBank database searches, the isolates causing burn spot disease symptoms were identified as Fusarium avenaceum in mainland Estonia and F. solani in Saaremaa crayfish.

  17. Detection of white spot syndrome virus (WSSV) of Penaeus chinensis by in situ hybridization

    NASA Astrophysics Data System (ADS)

    Zhan, Wen-Bin; Wang, Yuan-Hong; Zhang, Zhi-Dong; Hideo, Fukuda

    2000-09-01

    White Spot Syndrome Virus (WSSV) was purified from hemolymph of infected shrimp. After nucleic acid extraction from the purified virus particles, EcoR I-digested fragments of the WSSV genome were cloned; three of these fragments were used as non-radioactive probes labeled with DIG-11-dUTP. The probes hybridized in situ, with sections located in the nuclei of all WSSV-infected tissues. The virus was detected in the gill, stomach, epidermis, and connective tissue and so on, but not detected in healthy shrimp tissues and epithelial cells of hepatopancreatic tubules of diseased shrimp.

  18. The first case of Niikawa-Kuroki syndrome in Kazakhstan associated with café au lait spots.

    PubMed

    Al Mosawi, A J; Fewin, L

    2009-10-01

    Niikawa-Kuroki syndrome (Kabuki syndrome) is a multiple congenital anomaly syndrome of unknown etiology with a very wide spectrum of abnormalities and severity. The aim of this paper was to report the first case of the syndrome in Kazakhstan associated café au lait. Five year and half old boy from Kazakhstan (Uzbek-of Turk ethnicity) presented with dysmorphic facial features (long palpebral fissures, a broad and depressed nasal tip, large prominent earlobes, small head, epicanthic folds short stature, delayed language development, hypotonia, bilateral developmental dysplasia of the hip (DDH), large ears and triangular chin, café au lait spots. The clinical diagnosis was based on the triad of characteristic facial abnormalities (long palpebral fissures, a broad and depressed nasal tip, large prominent earlobes, small head), growth retardation, (DDH). In this paper the authors report the first case of Kabuki syndrome associated with café au lait spots.

  19. Arginine kinase of Litopenaeus vannamei involved in white spot syndrome virus infection.

    PubMed

    Ma, Fang-fang; Liu, Qing-hui; Guan, Guang-kuo; Li, Chen; Huang, Jie

    2014-04-10

    Virus-host interaction is important for virus infection. White spot syndrome virus VP14 contains transmembrane and signal peptides domain, which is considered to be important for virus infection. Until now, the function of this protein remains undefined. In this study, we explored the interaction of VP14 with host cell. A new shrimp protein (arginine kinase of Litopenaeus vannamei, LvAK) is selected and its localization in shrimp cells is also confirmed. Cellular localization of LvAK protein in shrimp hemocytes showed that LvAK was primarily located at the periphery of hemocytes and was scarcely detectable in the nucleus. Tissue distribution indicated that arginine kinase gene was spread commonly in the tissues and was highly present in shrimp muscle tissue. The expression of LvAK mRNA in muscle was significantly up-regulated after WSSV stimulation. Indirect immunofluorescence assay showed that LvAK interacted with VP14 in WSSV-infected shrimp. Injection of LvAK protein enhanced the mortality of shrimp infected with white spot syndrome virus (WSSV). These results showed that LvAK is involved in WSSV infection. Future research on this topic will help to reveal the molecular mechanism of WSSV infection.

  20. Multiple proteins of White spot syndrome virus involved in recognition of beta-integrin.

    PubMed

    Zhang, Jing-Yan; Liu, Qing-Hui; Huang, Jie

    2014-06-01

    The recognition and attachment of virus to its host cell surface is a critical step for viral infection. Recent research revealed that beta-integrin was involved in White spot syndrome virus (WSSV) infection. In this study, the interaction of beta-integrin with structure proteins of WSSV and motifs involved in WSSV infection was examined. The results showed that envelope proteins VP26, VP31, VP37, VP90 and nucleocapsid protein VP136 interacted with LvInt. RGD-, YGL- and LDV-related peptide functioned as motifs of WSSV proteins binding with beta-integrin. The beta-integrin ligand of RGDT had better blocking effect compared with that of YGL- and LDV-related peptides. In vivo assay indicated that RGD-, LDV- and YGL-related peptides could partially block WSSV infection. These data collectively indicate that multiple proteins were involved in recognition of beta-integrin. Identification of proteins in WSSV that are associated with beta-integrin will assist development of new agents for effective control of the white spot syndrome.

  1. Prevalence and distribution of White Spot Syndrome Virus in cultured shrimp.

    PubMed

    Hossain, A; Nandi, S P; Siddique, M A; Sanyal, S K; Sultana, M; Hossain, M A

    2015-02-01

    White Spot Syndrome Virus (WSSV) is a dsDNA virus causing White Spot Syndrome Disease (WSSD) in shrimp with almost 100% morality rate within 3-10 days. In Bangladesh, WSSD is one of the major impediments of shrimp farming. This study first investigated the prevalence and distribution of WSSV in cultured shrimps of the coastal regions in Bangladesh. A total of 60 shrimp samples, collected from the 25 shrimp farms of different coastal regions (Satkhira, Khulna, Bagerhat and Cox's Bazar), were analysed during 2013-2014 by conventional PCR using VP28 and VP664 gene-specific primers; 39 of 60 samples were found WSSV positive. SYBR green real-time PCR using 71-bp amplicon for VP664 gene correlated well with conventional PCR data. The prevalence rates of WSSV among the collected 60 samples were Satkhira 79%, Khulna 50%, Bagerhat 38% and Cox's Bazar 25%. Sequencing of WSSV-positive PCR amplicons of VP28 showed 99% similarity with WSSV NCBI Ref/Seq Sequences. Molecular analysis of the VP28 gene sequences of WSSV revealed that Bangladeshi strains phylogenetically affiliated to the strains belong to India. This work concluded that WSSV infections are widely distributed in the coastal regions cultured shrimp in Bangladesh.

  2. Expression, purification and crystallization of a novel nonstructural protein VP9 from white spot syndrome virus

    SciTech Connect

    Liu, Yang; Sivaraman, J.; Hew, Choy L.

    2006-08-01

    The nonstructural protein VP9 from white spot syndrome virus (WSSV) has been identified and expressed in Escherichia coli. Native protein was purified and crystallized by vapour diffusion. The nonstructural protein VP9 from white spot syndrome virus (WSSV) has been identified and expressed in Escherichia coli. To facilitate purification, a cleavable His{sub 6} tag was introduced at the N-terminus. The native protein was purified and crystallized by vapour diffusion against mother liquor containing 2 M sodium acetate, 100 mM MES pH 6.3, 25 mM cadmium sulfate and 3% glycerol. Crystals were obtained within 7 d and diffracted to 2.2 Å; they belonged to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 74.13, b = 78.21, c = 78.98 Å and four molecules in the asymmetric unit. The selenomethionine-labelled protein produced isomorphous crystals that diffracted to approximately 3.3 Å.

  3. [Relationship between spot urine pH and number of metabolic syndrome features].

    PubMed

    Yamanishi, Hachiro

    2014-07-01

    A significant inverse relationship between the 24-hour urine pH and number of metabolic syndrome (MS) features(BMI ≥ 30 kg/m2, circumference > 88cm, HDL-cholesterol < 50mg/dL, etc.) was identified in individuals, which means that 24-hour urine pH values decrease with an increasing number of MS features. In this respect, we examined the association between the number of MS features and spot urine pH instead of 24-hour urine pH. A positive relationship was observed between the spot urine pH and number of MS features by analysis of covariance, with the age, gender, and eGFR as covariates. On the other hand, a high level of serum uric acid (UA) indicated a significantly lower spot urine pH compared with a normal level of UA. Serum UA was a significant independent variable explaining the decrease of the urine pH. The results of path analysis showed that UA had negative effects on the number of MS features and urine pH. Therefore, a positive relationship between the number of MS features and urine pH will be a spurious correlation with UA as a confounding factor. In addition, the negative relationship between the number of MS features and UA was also a spurious correlation with age as a confounding factor. From these results, the serum UA level should be considered to evaluate the relationship between the number of MS features and spot urine pH.

  4. Crassostrea gigas oysters as a shrimp farm bioindicator of white spot syndrome virus.

    PubMed

    Vazquez-Boucard, C; Escobedo-Fregoso, C; Duran-Avelar, Ma de J; Mercier, L; Llera-Herrera, R; Escobedo-Bonilla, C; Vibanco-Perez, N

    2012-04-26

    This study explored whether Crassostrea gigas oysters can be used as a bioindicator of white spot syndrome virus (WSSV) in shrimp farm water canals. Bioassays showed that C. gigas can accumulate WSSV in their gills and digestive glands but do not become infected, either by exposure to seawater containing WSSV or by cohabitation with infected shrimp. The use of a WSSV nested PCR to screen oysters placed in water canals at the entry of a shrimp farm allowed WSSV to be detected 16 d prior to the disease occurring. The finding that C. gigas can concentrate small amounts of WSSV present in seawater without being harmed makes it an ideal sentinel species at shrimp farms.

  5. Optimized PCR assay for detection of white spot syndrome virus (WSSV).

    PubMed

    Nunan, Linda M; Lightner, Donald V

    2011-01-01

    A rapid PCR assay for detection of white spot syndrome virus (WSSV) was developed based on the nested PCR procedure described by Lo et al. (1996) and outlined as the recommended PCR diagnostic assay in the Manual of Diagnostic Tests for Aquatic Animals published by the Office of International Epizootics (OIE, 2009). The optimized procedure incorporated the second step primers used in the nested WSSV PCR. By adjusting the annealing temperature and shortening the cycling times, this modified assay is substantially faster and as sensitive as the recommended OIE protocol. The modified PCR test was compared directly to the two-step nested PCR protocol and a modified nested procedure. The sensitivity of the published assay was determined by template dilutions of semi-purified WSSV virions that had been quantitated using real-time PCR for detection of WSSV. Various isolates were tested using the modified procedure, to ensure that the assay was able to detect WSSV from different geographical locations.

  6. White spot syndrome virus strains of different virulence induce distinct immune response in Cherax quadricarinatus.

    PubMed

    Gao, Meiling; Li, Fang; Xu, Limei; Zhu, Xiaoming

    2014-07-01

    In this study, we identified three white spot syndrome virus (WSSV) strains (WSSV-CN01, WSSV-CN02 and WSSV-CN03) with significant differences in virulence. Among them, WSSV-CN01 caused significant higher and earlier mortality in redclaw crayfish Cherax quadricarinatus, thus was determined as high-virulent, while WSSV-CN02 and WSSV-CN03 were moderate-virulent and low-virulent. By investigating the total number of the circulating haemocytes and the activity of immune relative enzymes, we demonstrated that the different virulent WSSV strains induced distinct immune response in the host. Notably, a dramatic reduction of circulating haemocytes was observed in the crayfish infected with WSSV-CN01 and WSSV-CN02 but not WSSV-CN03. Further analysis revealed that cell death induced by WSSV-CN01 and WSSV-CN02 might be responsible for the decrease of circulating haemocytes.

  7. Endocytic pathway is indicated for white spot syndrome virus (WSSV) entry in shrimp.

    PubMed

    Huang, Zih-Jhan; Kang, Shih-Ting; Leu, Jiann-Horng; Chen, Li-Li

    2013-09-01

    The white spot syndrome virus (WSSV) has had a serious economic impact on the global shrimp aquaculture industry in the past two decades. Although research has clarified a lot about its genome and structure, the mechanism of how WSSV enters a cell is still unclear. In this study to determine this mechanism, primary cultured hemocytes were used as an experimental model to observe the process of WSSV entry because the stable shrimp cell lines for WSSV infection are lacking. After labeling virions and endosomes with fluorescent dyes followed by observation with a confocal microscope, the results show that the WSSV colocalizes with early endosomes. Hemocytes are further treated with different endocytic inhibitors, methyl-β-cyclodextrin (MβCD) and chlorpromazine (CPZ). WSSV still can be detected in the hemocytes treated with CPZ, but not in the hemocytes treated with MβCD. Thus, we conclude that WSSV adopts the caveolae-mediated endocytosis to enter the shrimp cell.

  8. Characterization of white spot syndrome virus VP52B and its interaction with VP26.

    PubMed

    Lin, Fanyu; Jie, Zuliang; Hou, Luhong; Li, Fang; Yang, Feng

    2015-02-01

    White spot syndrome virus (WSSV) is one of the major pathogens of cultured shrimp. Identification of envelope protein interactions has become a central issue for the understanding of WSSV assembly. In this paper, WSSV envelope protein VP52B was fused with GST-tag and expressed in Escherichia coli BL-21(DE3). Immunogold-electron microscopy revealed that VP52B was located on the outside surface of WSSV virions. Far-Western blotting analysis suggested that VP52B might directly interact with a major viral envelope protein VP26, and their interaction was confirmed by GST pull-down assay. Further investigation showed that the VP52B binding domain was located between residues 135-170 of VP26. These findings will enhance our understanding of the molecular mechanisms of WSSV morphogenesis.

  9. White spot syndrome virus enters crayfish hematopoietic tissue cells via clathrin-mediated endocytosis.

    PubMed

    Huang, Jiajun; Li, Fang; Wu, Junjun; Yang, Feng

    2015-12-01

    White spot syndrome virus (WSSV) is a major pathogen of aquacultured shrimp. However, the mechanism of its entry remains poorly understood. In this study, by analyzing the internalization of WSSV using crayfish hematopoietic tissue (HPT) cells, we showed that WSSV virions were engulfed by cell membrane invaginations sharing the features of clathrin-coated pits and then internalized into coated cytoplasmic vesicles. Further investigation indicated that WSSV internalization was significantly inhibited by chlorpromazine (CPZ) but not genistein. The internalized virions were colocalized with endogenous clathrin as well as transferrin which undergoes clathrin-dependent uptake. Preventing endosome acidification by ammonium chloride (NH4Cl) or chloroquine (CQ) dramatically reduced WSSV entry as well. Moreover, disturbance of dynamin activity or depletion of membrane cholesterol also blocked WSSV uptake. These data indicate that WSSV enters crayfish HPT cells via clathrin-mediated endocytosis in a pH-dependent manner, and membrane cholesterol as well as dynamin is critical for efficient viral entry.

  10. DNA condensates organized by the capsid protein VP15 in White Spot Syndrome Virus.

    PubMed

    Liu, Yingjie; Wu, Jinlu; Chen, Hu; Hew, Choy Leong; Yan, Jie

    2010-12-20

    The White Spot Syndrome Virus (WSSV) has a large circular double-stranded DNA genome of around 300kb and it replicates in the nucleus of the host cells. The machinery of how the viral DNA is packaged has been remained unclear. VP15, a highly basic protein, is one of the major capsid proteins found in the virus. Previously, it was shown to be a DNA binding protein and was hypothesized to participate in the viral DNA packaging process. Using Atomic Force Microscopy imaging, we show that the viral DNA is associated with a (or more) capsid proteins. The organized viral DNA qualitatively resembles the conformations of VP15 induced DNA condensates in vitro. Furthermore, single-DNA manipulation experiments revealed that VP15 is able to condense single DNA against forces of a few pico Newtons. Our results suggest that VP15 may aid in the viral DNA packaging process by directly condensing DNA.

  11. Binding of white spot syndrome virus to Artemia sp. cell membranes.

    PubMed

    Feng, Shuying; Li, Guangda; Feng, Wenpo; Huang, Jie

    2013-10-01

    Using differential velocity centrifugation, cell membranes of Artemia sp. were prepared, and their binding to white spot syndrome virus (WSSV) was analyzed in vitro. The results indicated that WSSV can specifically bind to Artemia cell membranes, and that WSSV receptor very likely existed in this membrane, which suggested that Artemia sp. may be a reservoir of WSSV. This study investigated the specific WSSV binding site by performing competitive inhibition experiments using shrimp gill cell membranes to bind WSSV to Artemia cell membranes. The results showed that shrimp gill cell membranes had a distinct inhibition effect on the specific binding of Artemia cell membranes to WSSV. Thus, potentially similar WSSV receptors or binding sites existed on Artemia sp. cell membranes and shrimp gill cell membranes. Taken together, these findings may provide experimental basis for the development of an effective approach to controlling WSSV, and theoretical basis for the study of WSSV receptors.

  12. Functional identification of the non-specific nuclease from white spot syndrome virus

    SciTech Connect

    Li Li; Lin Shumei; Yanga Feng . E-mail: mbiotech@public.xm.fj.cn

    2005-07-05

    The product encoded by the wsv191 gene from shrimp white spot syndrome virus (WSSV) is homologous with non-specific nucleases (NSN) of other organisms. To functionally identify the protein, the wsv191 gene was expressed in Escherichia coli as a glutathione S-transferase (GST) fusion protein with 6His-tag at C-terminal. The fusion protein (termed as rWSSV-NSN) was purified using Ni-NTA affinity chromatography under denatured conditions, renatured and characterized by three methods. The results showed that rWSSV-NSN could hydrolyze both DNA and RNA. 5'-RACE result revealed that the transcription initiation site of the wsv191 gene was located at nucleotide residue G of the predicted ATG triplet. Therefore, we concluded that the next ATG should be the genuine translation initiation codon of the wsv191 gene. Western blot analysis revealed that the molecular mass of natural WSSV-NSN was 37 kDa.

  13. The effects of a thermophile metabolite, tryptophol, upon protecting shrimp against white spot syndrome virus.

    PubMed

    Zhu, Fei; Jin, Min

    2015-12-01

    White spot syndrome virus (WSSV) is a shrimp pathogen responsible for significant economic loss in commercial shrimp farms and until now, there has been no effective approach to control this disease. In this study, tryptophol (indole-3-ethanol) was identified as a metabolite involved in bacteriophage-thermophile interactions. The dietary addition of tryptophol reduced the mortality in shrimp Marsupenaeus japonicus when orally challenged with WSSV. Our results revealed that 50 mg/kg tryptophol has a better protective effect in shrimp than 10 or 100 mg/kg tryptophol. WSSV copies in shrimp were reduced significantly (P < 0.01) when supplemented with 50 mg/kg tryptophol, indicating that virus replication was inhibited by tryptophol. Consequently, tryptophol represents an effective antiviral dietary supplement for shrimp, and thus holds significant promise as a novel and efficient therapeutic approach to control WSSV in shrimp aquaculture.

  14. Rapid detection of shrimp white spot syndrome virus by real time, isothermal recombinase polymerase amplification assay.

    PubMed

    Xia, Xiaoming; Yu, Yongxin; Weidmann, Manfred; Pan, Yingjie; Yan, Shuling; Wang, Yongjie

    2014-01-01

    White spot syndrome virus (WSSV) causes large economic losses to the shrimp aquaculture industry, and thus far there are no efficient therapeutic treatments available against this lethal virus. In this study, we present the development of a novel real time isothermal recombinase polymerase amplification (RPA) assay for WSSV detection on a small ESEQuant Tube Scanner device. The RPA sensitivity, specificity and rapidity were evaluated by using a plasmid standard as well as viral and shrimp genomic DNAs. Compared with qPCR, the RPA assay revealed more satisfactory performance. It reached a detection limit up to 10 molecules in 95% of cases as determined by probit analysis of 8 independent experiments within 6.41 ± 0.17 min at 39 °C. Consequently, this rapid RPA method has great application potential for field use or point of care diagnostics.

  15. Expression, purification and crystallization of a novel nonstructural protein VP9 from white spot syndrome virus.

    SciTech Connect

    Liu,Y.; Sivaraman, J.; Hew, C.

    2006-01-01

    The nonstructural protein VP9 from white spot syndrome virus (WSSV) has been identified and expressed in Escherichia coli. To facilitate purification, a cleavable His{sub 6} tag was introduced at the N-terminus. The native protein was purified and crystallized by vapor diffusion against mother liquor containing 2 M sodium acetate, 100 mM MES pH 6.3, 25 mM cadmium sulfate and 3% glycerol. Crystals were obtained within 7 d and diffracted to 2.2 Angstroms; they belonged to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 74.13, b = 78.21, c = 78.98 Angstroms and four molecules in the asymmetric unit. The selenomethionine-labeled protein produced isomorphous crystals that diffracted to approximately 3.3 Angstroms.

  16. Rett syndrome in females with CTS hot spot deletions: a disorder profile.

    PubMed

    Smeets, E; Terhal, P; Casaer, P; Peters, A; Midro, A; Schollen, E; van Roozendaal, K; Moog, U; Matthijs, G; Herbergs, J; Smeets, H; Curfs, L; Schrander-Stumpel, C; Fryns, J P

    2005-01-15

    From a series of 107 females with Rett syndrome (RTT), we describe the long-term history of ten females with a deletion in the C-terminus of the MECP2 gene. We observed that their disorder profile is clinically recognizable with time and different from other atypical and milder RTT phenotypes. In females with hot spot deletions in the C-terminus, dystonia is present from childhood and results in a serious spine deformation in spite of preventive measures. Their adaptive behavior is surprisingly better preserved and in contrast with the typical decline in motor functioning. The delineation of disorder profiles by long-term clinical observation can teach us about genotype/phenotype relationships and eventually about the effect of epigenetic phenomena on the final phenotype.

  17. Molecular Mechanisms of White Spot Syndrome Virus Infection and Perspectives on Treatments.

    PubMed

    Verbruggen, Bas; Bickley, Lisa K; van Aerle, Ronny; Bateman, Kelly S; Stentiford, Grant D; Santos, Eduarda M; Tyler, Charles R

    2016-01-18

    Since its emergence in the 1990s, White Spot Disease (WSD) has had major economic and societal impact in the crustacean aquaculture sector. Over the years shrimp farming alone has experienced billion dollar losses through WSD. The disease is caused by the White Spot Syndrome Virus (WSSV), a large dsDNA virus and the only member of the Nimaviridae family. Susceptibility to WSSV in a wide range of crustacean hosts makes it a major risk factor in the translocation of live animals and in commodity products. Currently there are no effective treatments for this disease. Understanding the molecular basis of disease processes has contributed significantly to the treatment of many human and animal pathogens, and with a similar aim considerable efforts have been directed towards understanding host-pathogen molecular interactions for WSD. Work on the molecular mechanisms of pathogenesis in aquatic crustaceans has been restricted by a lack of sequenced and annotated genomes for host species. Nevertheless, some of the key host-pathogen interactions have been established: between viral envelope proteins and host cell receptors at initiation of infection, involvement of various immune system pathways in response to WSSV, and the roles of various host and virus miRNAs in mitigation or progression of disease. Despite these advances, many fundamental knowledge gaps remain; for example, the roles of the majority of WSSV proteins are still unknown. In this review we assess current knowledge of how WSSV infects and replicates in its host, and critique strategies for WSD treatment.

  18. White spot syndrome virus epizootic in cultured Pacific white shrimp Litopenaeus vannamei (Boone) in Taiwan.

    PubMed

    Cheng, L; Lin, W-H; Wang, P-C; Tsai, M-A; Hsu, J-P; Chen, S-C

    2013-12-01

    White spot syndrome virus (WSSV) has caused significant losses in shrimp farms worldwide. Between 2004 and 2006, Pacific white shrimp Litopenaeus vannamei (Boone) were collected from 220 farms in Taiwan to determine the prevalence and impact of WSSV infection on the shrimp farm industry. Polymerase chain reaction (PCR) analysis detected WSSV in shrimp from 26% of farms. Juvenile shrimp farms had the highest infection levels (38%; 19/50 farms) and brooder shrimp farms had the lowest (5%; one of 20 farms). The average extent of infection at each farm was as follows for WSSV-positive farms: post-larvae farms, 71%; juvenile farms, 61%; subadult farms, 62%; adult farms, 49%; and brooder farms, 40%. Characteristic white spots, hypertrophied nuclei and basophilic viral inclusion bodies were found in the epithelia of gills and tail fans, appendages, cephalothorax and hepatopancreas, and virions of WSSV were observed. Of shrimp that had WSSV lesions, 100% had lesions on the cephalothorax, 96% in gills and tail fans, 91% on appendages and 17% in the hepatopancreas. WSSV was also detected in copepoda and crustaceans from the shrimp farms. Sequence comparison using the pms146 gene fragment of WSSV showed that isolates from the farms had 99.7-100% nucleotide sequence identity with four strains in the GenBank database--China (AF332093), Taiwan (AF440570 and U50923) and Thailand (AF369029). This is the first broad study of WSSV infection in L. vannamei in Taiwan.

  19. Molecular Mechanisms of White Spot Syndrome Virus Infection and Perspectives on Treatments

    PubMed Central

    Verbruggen, Bas; Bickley, Lisa K.; van Aerle, Ronny; Bateman, Kelly S.; Stentiford, Grant D.; Santos, Eduarda M.; Tyler, Charles R.

    2016-01-01

    Since its emergence in the 1990s, White Spot Disease (WSD) has had major economic and societal impact in the crustacean aquaculture sector. Over the years shrimp farming alone has experienced billion dollar losses through WSD. The disease is caused by the White Spot Syndrome Virus (WSSV), a large dsDNA virus and the only member of the Nimaviridae family. Susceptibility to WSSV in a wide range of crustacean hosts makes it a major risk factor in the translocation of live animals and in commodity products. Currently there are no effective treatments for this disease. Understanding the molecular basis of disease processes has contributed significantly to the treatment of many human and animal pathogens, and with a similar aim considerable efforts have been directed towards understanding host–pathogen molecular interactions for WSD. Work on the molecular mechanisms of pathogenesis in aquatic crustaceans has been restricted by a lack of sequenced and annotated genomes for host species. Nevertheless, some of the key host–pathogen interactions have been established: between viral envelope proteins and host cell receptors at initiation of infection, involvement of various immune system pathways in response to WSSV, and the roles of various host and virus miRNAs in mitigation or progression of disease. Despite these advances, many fundamental knowledge gaps remain; for example, the roles of the majority of WSSV proteins are still unknown. In this review we assess current knowledge of how WSSV infects and replicates in its host, and critique strategies for WSD treatment. PMID:26797629

  20. Expression, purification and crystallization of two major envelope proteins from white spot syndrome virus

    SciTech Connect

    Tang, Xuhua; Hew, Choy Leong

    2007-07-01

    The crystallization of the N-terminal transmembrane region-truncated VP26 and VP28 of white spot syndrome virus is described. White spot syndrome virus (WSSV) is a major virulent pathogen known to infect penaeid shrimp and other crustaceans. VP26 and VP28, two major envelope proteins from WSSV, have been identified and overexpressed in Escherichia coli. In order to facilitate purification and crystallization, predicted N-terminal transmembrane regions of approximately 35 amino acids have been truncated from both VP26 and VP28. Truncated VP26 and VP28 and their corresponding SeMet-labelled proteins were purified and the SeMet proteins were crystallized by the hanging-drop vapour-diffusion method. Crystals of SeMet-labelled VP26 were obtained using a reservoir consisting of 0.1 M citric acid pH 3.5, 3.0 M sodium chloride and 1%(w/v) polyethylene glycol 3350, whereas SeMet VP28 was crystallized using a reservoir solution consisting of 25% polyethylene glycol 8000, 0.2 M calcium acetate, 0.1 M Na HEPES pH 7.5 and 1.5%(w/v) 1,2,3-heptanetriol. Crystals of SeMet-labelled VP26 diffract to 2.2 Å resolution and belong to space group R32, with unit-cell parameters a = b = 73.92, c = 199.31 Å. SeMet-labelled VP28 crystallizes in space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 105.33, b = 106.71, c = 200.37 Å, and diffracts to 2.0 Å resolution.

  1. Field-Usable Lateral Flow Immunoassay for the Rapid Detection of White Spot Syndrome Virus (WSSV)

    PubMed Central

    Kulabhusan, Prabir Kumar; Rajwade, Jyutika M.; Sugumar, Vimal; Taju, Gani; Sahul Hameed, A. S.

    2017-01-01

    Background White spot disease (WSD), a major threat to sustainable aquaculture worldwide, is caused by White spot syndrome virus (WSSV). The diagnosis of WSD relies heavily on molecular detection of the virus by one-step PCR. These procedures are neither field-usable nor rapid enough considering the speed at which the virus spreads. Thus, development of a rapid, reliable and field-usable diagnostic method for the detection of WSSV infection is imperative to prevent huge economic losses. Methods/Principal Findings Here, we report on the development of a lateral flow immunoassay (LFIA) employing gold nanoparticles conjugated to a polyclonal antibody against VP28 (envelope protein of WSSV). The LFIA detected WSSV in ~20 min and showed no cross-reactivity with other shrimp viruses, viz. Monodon Baculovirus (MBV), Hepatopancreatic parvovirus (HPV) and Infectious Hypodermal and Hematopoietic Necrosis virus (IHHNV). The limit of detection (LOD) of the assay, as determined by real-time PCR, was 103 copies of WSSV. In a time course infectivity experiment, ~104 WSSV particles were injected in Litopenaeus vannamei. The LFIA could rapidly (~ 20 min) detect the virus in different tissues after 3 h (hemolymph), 6 h (gill tissue) and 12 h (head soft tissue, eye stalk, and pleopod) of infection. Based on these findings, a validation study was performed using 75 field samples collected from different geographical locations in India. The LFIA results obtained were compared with the conventional “gold standard test”, viz. one-step PCR. The analysis of results in 2x2 matrix indicated very high sensitivity (100%) and specificity (96.77%) of LFIA. Similarly, Cohen’s kappa coefficient of 0.983 suggested "very good agreement” between the developed LFIA and the conventional one-step PCR. Conclusion The LFIA developed for the rapid detection of WSSV has an excellent potential for use in the field and could prove to be a boon to the aquaculture industry. PMID:28046005

  2. Hijacking of host calreticulin is required for the white spot syndrome virus replication cycle.

    PubMed

    Watthanasurorot, Apiruck; Guo, Enen; Tharntada, Sirinit; Lo, Chu-Fang; Söderhäll, Kenneth; Söderhäll, Irene

    2014-07-01

    We have previously shown that multifunctional calreticulin (CRT), which resides in the endoplasmic reticulum (ER) and is involved in ER-associated protein processing, responds to infection with white spot syndrome virus (WSSV) by increasing mRNA and protein expression and by forming a complex with gC1qR and thereby delaying apoptosis. Here, we show that CRT can directly interact with WSSV structural proteins, including VP15 and VP28, during an early stage of virus infection. The binding of VP28 with CRT does not promote WSSV entry, and CRT-VP15 interaction was detected in the viral genome in virally infected host cells and thus may have an effect on WSSV replication. Moreover, CRT was detected in the viral envelope of purified WSSV virions. CRT was also found to be of high importance for proper oligomerization of the viral structural proteins VP26 and VP28, and when CRT glycosylation was blocked with tunicamycin, a significant decrease in both viral replication and assembly was detected. Together, these findings suggest that CRT confers several advantages to WSSV, from the initial steps of WSSV infection to the assembly of virions. Therefore, CRT is required as a "vital factor" and is hijacked by WSSV for its replication cycle. Importance: White spot syndrome virus (WSSV) is a double-stranded DNA virus and the cause of a serious disease in a wide range of crustaceans that often leads to high mortality rates. We have previously shown that the protein calreticulin (CRT), which resides in the endoplasmic reticulum (ER) of the cell, is important in the host response to the virus. In this report, we show that the virus uses this host protein to enter the cell and to make the host produce new viral structural proteins. Through its interaction with two viral proteins, the virus "hijacks" host calreticulin and uses it for its own needs. These findings provide new insight into the interaction between a large DNA virus and the host protein CRT and may help in understanding

  3. BAX inhibitor-1 silencing suppresses white spot syndrome virus replication in red swamp crayfish, Procambarus clarkii.

    PubMed

    Du, Zhi-Qiang; Lan, Jiang-Feng; Weng, Yu-Ding; Zhao, Xiao-Fan; Wang, Jin-Xing

    2013-07-01

    BAX inhibitor-1 (BI-1) was originally described as an anti-apoptotic protein in both animal and plant cells. BI-1 overexpression suppresses ER stress-induced apoptosis in animal cells. Inhibition of BI-1 activity could induce the cell death in mammals and plants. However, the function of BI-1 in crustacean immunity was unclear. In this paper, the full-length cDNA of a BI-1 protein in red swamp crayfish, Procambarus clarkii (PcBI-1) was cloned and its expression profiles in normal and infected crayfish were analyzed. The results showed that PcBI-1 was expressed in hemocytes, heart, hepatopancreas, gills, stomach, and intestines of the crayfish and was upregulated after challenged with Vibrio anguillarum and with white spot syndrome virus (WSSV). To determine the function of PcBI-1 in the innate immunity of the crayfish, the RNA interference against PcBI-1 was performed and the results indicated the hemocyte programmed cell death rate was increased significantly and WSSV replication was declined after PcBI-1 knocked down. Altogether, PcBI-1 plays an anti-apoptotic role, wherein high PcBI-1 expression suppresses programmed cell death, which is beneficial for WSSW replication in crayfish.

  4. Viral ubiquitin ligase WSSV222 is required for efficient white spot syndrome virus replication in shrimp.

    PubMed

    He, Fang; Syed, Syed Musthaq; Hameed, A S Sahul; Kwang, Jimmy

    2009-06-01

    The E3 ligase WSSV222 of white spot syndrome virus (WSSV) is involved in anti-apoptosis regulation by ubiquitin-mediated degradation of tumour suppressor-like protein (TSL), a shrimp tumour suppressor. In the present study, WSSV222 gene expression was silenced by using specific small interfering RNA (siRNA) in Sf9 and BHK cells. Based on the results of the in vitro silencing, WSSV-challenged shrimp were treated with anti-WSSV222 siRNA to knock down WSSV222 protein expression. The survival rate of shrimp and the efficiency of WSSV replication were assessed to evaluate the efficacy of anti-WSSV222 siRNA in regulating WSSV infection in shrimp. The anti-WSSV222 siRNA reduced the cumulative mortality in shrimp challenged with 10(3) copies of WSSV and delayed the mean time to death in shrimp challenged with the higher dose of 10(6) copies. The results of real-time quantitative PCR showed that virus replication was delayed and reduced in WSSV-challenged shrimp treated with anti-WSSV222 siRNA in comparison with challenged shrimp treated with random-control siRNA. Co-immunoprecipitation assays revealed that WSSV222 silencing inhibited the degradation of TSL in WSSV-challenged shrimp, indicating the requirement for WSSV222 for efficient replication of WSSV in shrimp.

  5. Laminin Receptor in Shrimp Is a Cellular Attachment Receptor for White Spot Syndrome Virus

    PubMed Central

    Liu, Wang-Jing; Li, Yi-Chieh; Kou, Guang-Hsiung; Lo, Chu-Fang

    2016-01-01

    White spot syndrome virus (WSSV, genus Whispovirus, family Nimaviridae) is causing huge economic losses in global shrimp farming, but there is no effective control. Shrimp cell laminin receptor (Lamr) may have a role in WSSV infection. The objective was to characterize interactions between Penaeus monodon Lamr (PmLamr) and WSSV structural proteins. In this study, PmLamr interacted with nine WSSV structural proteins (based on yeast two-hybrid screening), of which one (VP31) was characterized. Protein pull-down assay confirmed the interaction between PmLamr and VP31; the latter was an envelope protein exposed outside the WSSV virion (based on membrane topology assays). Furthermore, similar to mammalian Lamr, there were two major protein bands in shrimp cells. Cellular localization assay demonstrated VP31 co-localized with PmLamr on transfected cells. Enzyme-link immunosorbent assay (ELISA) and competitive ELISA demonstrated binding of VP31 on PmLamr was dose-dependent; however, addition of WSSV virion competed for binding affinity. Furthermore, based on an in vivo neutralization assay, both VP31 and PmLamr delayed mortality in shrimp challenged with WSSV. We concluded Lamr was an important receptor for WSSV infection and the viral envelope protein VP31 may have a role in host cell recognition and binding. These data contributed to elucidating pathogenesis of WSSV infection and may help in controlling this disease. PMID:27257954

  6. Laminin Receptor in Shrimp Is a Cellular Attachment Receptor for White Spot Syndrome Virus.

    PubMed

    Liu, Wang-Jing; Li, Yi-Chieh; Kou, Guang-Hsiung; Lo, Chu-Fang

    2016-01-01

    White spot syndrome virus (WSSV, genus Whispovirus, family Nimaviridae) is causing huge economic losses in global shrimp farming, but there is no effective control. Shrimp cell laminin receptor (Lamr) may have a role in WSSV infection. The objective was to characterize interactions between Penaeus monodon Lamr (PmLamr) and WSSV structural proteins. In this study, PmLamr interacted with nine WSSV structural proteins (based on yeast two-hybrid screening), of which one (VP31) was characterized. Protein pull-down assay confirmed the interaction between PmLamr and VP31; the latter was an envelope protein exposed outside the WSSV virion (based on membrane topology assays). Furthermore, similar to mammalian Lamr, there were two major protein bands in shrimp cells. Cellular localization assay demonstrated VP31 co-localized with PmLamr on transfected cells. Enzyme-link immunosorbent assay (ELISA) and competitive ELISA demonstrated binding of VP31 on PmLamr was dose-dependent; however, addition of WSSV virion competed for binding affinity. Furthermore, based on an in vivo neutralization assay, both VP31 and PmLamr delayed mortality in shrimp challenged with WSSV. We concluded Lamr was an important receptor for WSSV infection and the viral envelope protein VP31 may have a role in host cell recognition and binding. These data contributed to elucidating pathogenesis of WSSV infection and may help in controlling this disease.

  7. White spot syndrome virus (WSSV) infection in shrimp (Litopenaeus vannamei) exposed to low and high salinity.

    PubMed

    Ramos-Carreño, Santiago; Valencia-Yáñez, Ricardo; Correa-Sandoval, Francisco; Ruíz-García, Noé; Díaz-Herrera, Fernando; Giffard-Mena, Ivone

    2014-09-01

    White spot syndrome virus (WSSV) has a worldwide distribution and is considered one of the most pathogenic and devastating viruses to the shrimp industry. A few studies have explored the effect of WSSV on shrimp acclimated to low (5 practical salinity units [psu]) or high ([40 psu) salinity conditions. In this work, we analysed the physiological response of WSSV-infected Litopenaeus vannamei juveniles that were acclimated to different salinities (5, 15, 28, 34 and 54 psu). We evaluated the osmotic response and survival of the shrimp at different times after infection (0 to 48 hours), and we followed the expression levels of a viral gene (vp664) in shrimp haemolymph using real-time PCR. Our results indicate that the susceptibility of the shrimp to the virus increased at extreme salinities (5 and 54 psu), with higher survival rates at 15 and 28 psu, which were closer to the iso-osmotic point (24.7 psu, 727.5 mOsmol/kg). Acute exposure to the virus made the haemolymph less hyperosmotic at 5 and 15 psu and less hypo-osmotic at higher salinities ([28 psu). The capacity of white shrimp to osmoregulate, and thus survive, significantly decreased following WSSV infection. According to our results, extreme salinities (5 or 54 psu) are more harmful than seawater.

  8. Crystal Structure of Major Envelope Protein VP24 from White Spot Syndrome Virus

    PubMed Central

    Sun, Lifang; Su, Yintao; Zhao, Yanhe; Fu, Zheng-qing; Wu, Yunkun

    2016-01-01

    White spot syndrome virus (WSSV) is one of the major and most serious pathogen in the shrimp industry. As one of the most abundant envelope protein, VP24 acts as a core protein interacting with other structure proteins and plays an important role in virus assembly and infection. Here, we have presented the crystal structure of VP24 from WSSV. In the structure, VP24 consists of a nine-stranded β–barrel fold with mostly antiparallel β-strands, and the loops extending out the β–barrel at both N-terminus and C-terminus, which is distinct to those of the other two major envelope proteins VP28 and VP26. Structural comparison of VP24 with VP26 and VP28 reveals opposite electrostatic surface potential properties of them. These structural differences could provide insight into their differential functional mechanisms and roles for virus assembly and infection. Moreover, the structure reveals a trimeric assembly, suggesting a likely natural conformation of VP24 in viral envelope. Therefore, in addition to confirming the evolutionary relationship among the three abundant envelope proteins of WSSV, our structural studies also facilitate a better understanding of the molecular mechanism underlying special roles of VP24 in WSSV assembly and infection. PMID:27572278

  9. ICP35 Is a TREX-Like Protein Identified in White Spot Syndrome Virus

    PubMed Central

    Phairoh, Panapat; Suthibatpong, Thana; Rattanarojpong, Triwit; Jongruja, Nujarin; Senapin, Saengchan; Choowongkomon, Kiattawee; Khunrae, Pongsak

    2016-01-01

    ICP35 is a non-structural protein from White spot syndrome virus believed to be important in viral replication. Since ICP35 was found to localize in the host nucleus, it has been speculated that the function of ICP35 might be involved in the interaction of DNA. In this study, we overexpressed, purified and characterized ICP35. The thioredoxin-fused ICP35 (thio-ICP35) was strongly expressed in E. coli and be able to form itself into dimers. Investigation of the interaction between ICP35 and DNA revealed that ICP35 can perform DNase activity. Structural model of ICP35 was successfully built on TREX1, suggesting that ICP35 might adopt the folding similar to that of TREX1 protein. Several residues important for dimerization in TREX1 are also conserved in ICP35. Residue Asn126 and Asp132, which are seen to be in close proximity to metal ions in the ICP35 model, were shown through site-directed mutagenesis to be critical for DNase activity. PMID:27348862

  10. OIE white spot syndrome virus PCR gives false-positive results in Cherax quadricarinatus.

    PubMed

    Claydon, Kerry; Cullen, Bradford; Owens, Leigh

    2004-12-13

    White spot syndrome virus (WSSV) is an intranuclear bacilliform virus (IBV) that is a serious, notifiable crustacean pathogen. The Office International des Epizooties (OIE) PCR protocol for WSSV uses primer sets initially developed by Lo et al. (1996). It yields a first-step PCR amplicon of 1441 bp and a nested PCR amplicon of 941 bp. An amplicon (941 bp) purported to specifically detect WSSV was obtained when using template DNA extracted from Cherax quadricarinatus in a WSSV PCR detection protocol recommended by the OIE. Sequencing and analysis of the 941 bp amplicon and an occasional 550 bp amplicon from C. quadricarinatus revealed no phylogenetic relationship with WSSV, and suggested a possible lack of sufficient primer specificity for WSSV in the OIE test. This suggestion was supported by the fact that the OIE outer primer sequence (146F1) was present in both the forward and reverse position of the 941 bp and the forward position of the 550 bp nested amplicons from C. quadricarinatus. As WSSV is a notifiable pathogen, the consequences of false-positive results are harsh in WSSV-free zones and can lead to incorrect quarantine and unnecessary destruction of animals. Therefore, urgent attention and revision is necessary for the current OIE PCR protocol for WSSV detection.

  11. Fusion of flagellin 2 with bivalent white spot syndrome virus vaccine increases survival in freshwater shrimp.

    PubMed

    Cho, Hansam; Park, Na Hye; Jang, Yuyeon; Gwon, Yong-Dae; Cho, Yeondong; Heo, Yoon-Ki; Park, Ki-Hoon; Lee, Hee-Jung; Choi, Tae Jin; Kim, Young Bong

    2017-03-01

    Despite large economic losses attributable to white spot syndrome virus (WSSV), an infectious pathogen of penaeid shrimp and other crustaceans worldwide, no efficient vaccines or antiviral agents to control the virus are available at present. Here, we designed and constructed baculovirus-based vaccines delivering genes encoding the WSSV envelope proteins, VP28 and VP19. To enhance the immunogenicity of the baculovirus-based vaccine, we fused a Salmonella typhimurium flagellin 2 (FL2) gene with VP28 or VP19 gene. Both vaccine constructs elicited similar high titlers of anti-WSSV IgG after oral immunization in mice. The protective effect of oral vaccines upon WSSV challenge was observed in Macrobrachium nipponense. Bivalent vaccine displaying WSSV envelope proteins, VP19 and VP28, led to enhanced more than 10% survival protection against WSSV infection, compared to monovalent vaccine containing WSSV envelope protein, VP19 or VP28. Furthermore, a baculovirus-based WSSV vaccine fused with FL2 gene, Ac-VP28-ie1VP19FL2, efficiently protected mice against WSSV challenge (89.5% survival rate). In support of the efficacy of FL2 in our vaccine, we verified FL2 enhanced survival rate and induced the NF-κB gene in Palaemon paucidens. The collective results strongly suggest that our recombinant baculoviral system displaying WSSV envelope protein and delivering FL2-fused WSSV envelope gene effectively induced protective responses, supporting the utility of a potential new oral DNA vaccine against WSSV.

  12. Molecular immune response of the American lobster (Homarus americanus) to the White Spot Syndrome Virus.

    PubMed

    Clark, K Fraser; Greenwood, Spencer J; Acorn, Adam R; Byrne, Philip J

    2013-11-01

    The adult American lobster (Homarus americanus) is susceptible to few naturally occurring pathogens, and no viral pathogen is known to exist. Despite this, relatively little is known about the H. americanus immune system and nothing is known about its potential viral immune response. Hundreds of rural communities in Atlantic Canada rely on the lobster fishery for their economic sustainability and could be devastated by large-scale pathogen-mediated mortality events. The White Spot Syndrome Virus is the most economically devastating viral pathogen to global shrimp aquaculture production and has been proposed to be capable of infecting all decapod crustaceans including the European Lobster. An in vivo WSSV injection challenge was conducted in H. americanus and WSSV was found to be capable of infecting and replicating within lobsters held at 20°C. The in vivo WSSV challenge also generated the first viral disease model of H. americanus and allowed for the high-throughput examination of transcriptomic changes that occur during viral infection. Microarray analysis found 136 differentially expressed genes and the expression of a subset of these genes was verified using RT-qPCR. Anti-lipopolysaccharide isoforms and acute phase serum amyloid protein A expression did not change during WSSV infection, contrary to previous findings during bacterial and parasitic infection of H. americanus. This, along with the differential gene expression of thioredoxin and trypsin isoforms, provides compelling evidence that H. americanus is capable of mounting an immune response specific to infection by different pathogen classes.

  13. Hematological changes in white spot syndrome virus-infected shrimp, Fenneropenaeus chinensis (Osbeck)

    NASA Astrophysics Data System (ADS)

    Feng, Shouming; Zhan, Wenbin; Xing, Jing; Li, Jun; Yang, Kai; Wang, Jing

    2008-08-01

    The pathological changes of hemocytes in the haemolymph and hepatopancreas were examined in experimentally and naturally WSSV (white spot syndrome virus) infected Fenneropenaeus chinensis. The results showed that the pathological manifestations of hemocytes were similar among moribund shrimps infected via injection, feeding and by nature. Firstly, the total hemocyte counts (THCs) in WSSV-infected shrimp were significantly lower than those in healthy shrimp. Secondly, necrotic, broken and disintegrated cells were often observed, and a typical hematolysis was present in the haemolymph smear of WSSV-infected shrimp. Thirdly, necrosis and typical apoptosis of hemocytes were detected with TEM in the peripheral haemolymph of WSSV-infected shrimp. Hyalinocytes and semi-granulocytes with masses of WSSVs in their nuclei often appeared, whereas no granular hemocytes with WSSV were found in the hepatopancreas of moribund infected shrimps. All our results supported that hemocytes were the main target cells of WSSV, and hyalinocytes and semigranular hemocytes seemed to be more favorable for WSSV infection in F. chinensis.

  14. Characterization and interactome study of white spot syndrome virus envelope protein VP11.

    PubMed

    Liu, Wang-Jing; Shiung, Hui-Jui; Lo, Chu-Fang; Leu, Jiann-Horng; Lai, Ying-Jang; Lee, Tai-Lin; Huang, Wei-Tung; Kou, Guang-Hsiung; Chang, Yun-Shiang

    2014-01-01

    White spot syndrome virus (WSSV) is a large enveloped virus. The WSSV viral particle consists of three structural layers that surround its core DNA: an outer envelope, a tegument and a nucleocapsid. Here we characterize the WSSV structural protein VP11 (WSSV394, GenBank accession number AF440570), and use an interactome approach to analyze the possible associations between this protein and an array of other WSSV and host proteins. Temporal transcription analysis showed that vp11 is an early gene. Western blot hybridization of the intact viral particles and fractionation of the viral components, and immunoelectron microscopy showed that VP11 is an envelope protein. Membrane topology software predicted VP11 to be a type of transmembrane protein with a highly hydrophobic transmembrane domain at its N-terminal. Based on an immunofluorescence assay performed on VP11-transfected Sf9 cells and a trypsin digestion analysis of the virion, we conclude that, contrary to topology software prediction, the C-terminal of this protein is in fact inside the virion. Yeast two-hybrid screening combined with co-immunoprecipitation assays found that VP11 directly interacted with at least 12 other WSSV structural proteins as well as itself. An oligomerization assay further showed that VP11 could form dimers. VP11 is also the first reported WSSV structural protein to interact with the major nucleocapsid protein VP664.

  15. Toll receptor response to white spot syndrome virus challenge in giant freshwater prawns (Macrobrachium rosenbergii).

    PubMed

    Feng, Jinling; Zhao, Lingling; Jin, Min; Li, Tingting; Wu, Lei; Chen, Yihong; Ren, Qian

    2016-10-01

    Toll receptors are evolutionary ancient families of pattern recognition receptors with crucial roles in invertebrate innate immune response. In this study, we identified a Toll receptor (MrToll) from giant freshwater prawns (Macrobrachium rosenbergii). The full-length cDNA of MrToll is 4257 bp, which encodes a putative protein of 1367 amino acids. MrToll contains 17 LRR domains, a transmembrane domain, and a TIR domain. Phylogenetic analysis showed that MrToll was grouped with Drosophila Toll7 and other arthropod Tolls. The transcripts of MrToll are mainly distributed in the heart, hepatopancreas, gills, stomach, and intestine. A low level of MrToll expression can be detected in hemocytes and the lymphoid organ. MrToll expression in gills was gradually upregulated to the highest level from 24 h to 48 h during the white spot syndrome virus (WSSV) challenge. The expression levels of the crustin (Cru) genes Cru3 and Cru7 in gills were relatively lower than those of Cru2 and Cru4. The expression levels of Cru3 and Cru7 were inhibited after the RNA interference of MrToll in gills during the WSSV challenge. The anti-lipopolysaccharide factor (ALF) genes ALF2, ALF3, ALF4, and ALF5 were also regulated by MrToll in gills during the virus challenge. These findings suggest that MrToll may contribute to the innate immune defense of M. rosenbergii against WSSV.

  16. The role of white spot syndrome virus (WSSV) VP466 protein in shrimp antiviral phagocytosis.

    PubMed

    Ye, Ting; Zong, Rongrong; Zhang, Xiaobo

    2012-08-01

    Widespread evidence indicates that the structural proteins of virus play very important roles in virus-host interactions. However, the effect of viral proteins on host immunity has not been addressed. Our previous studies revealed that the host shrimp Rab6 (termed as PjRab previously), tropomyosin, β-actin and the white spot syndrome virus (WSSV) envelope protein VP466 formed a complex. In this study, the VP466 protein was shown to be able to bind host Rab6 protein and increase its GTPase activity in vivo and vitro. Thus, VP466 could function as a GTPase-activating protein (GAP) of Rab6. In the VP466-Rab-actin pathway, the increase of the Rab6 activity induced rearrangements of the actin cytoskeleton, resulting in the formation of actin stress fibers which promoted the phagocytosis against virus. Therefore our findings revealed that a viral protein could be employed by host to initiate the host immunity, representing a novel molecular mechanism in the virus-host interaction. Our study would help to better understand the molecular events in immune response against virus infection in invertebrates.

  17. Development of an immuno-based colorimetric assay for white spot syndrome virus.

    PubMed

    Loyprasert-Thananimit, Suchera; Saleedang, Akrapon; Deachamag, Panchalika; Waiyapoka, Thanyaporn; Neulplub, Maitee; Chotigeat, Wilaiwan

    2014-01-01

    White spot syndrome virus (WSSV) is a major cause of infectious disease in cultured shrimp. A fast and reliable method for detecting and monitoring the amount of WSSV during farming would be extremely useful. This work describes a sandwich immunoassay that uses anti-GST-VP26, a WSSV-binding protein (WBP), and modified streptavidin magnesphere paramagnetic particles (SMPPs) to develop the technique. The WBP was immobilized on SMPPs and later bound to different copies of WSSV. The binding was detected using anti-GST-VP26 conjugated to alkaline phosphatase. This enzymatic reaction successfully changed the test solution to a concentration-dependent yellow color that was measured at 405 nm. The sensitivity of this method was between 1.6 × 10(4) and 1.6 × 10(7) copies µL(-1) of WSSV. In this study, the color for detection and semiquantitative analysis is easily observed and measured and can lead to the development of a test kit for screening WSSV during shrimp farming.

  18. Envelope protein VP24 from White spot syndrome virus: expression, purification and crystallization.

    PubMed

    Sun, Lifang; Wu, Yunkun

    2016-08-01

    White spot syndrome virus (WSSV) is a major shrimp pathogen known to infect penaeid shrimp and other crustaceans. VP24 is one of the major envelope proteins of WSSV. In order to facilitate purification, crystallization and structure determination, the predicted N-terminal transmembrane region of approximately 26 amino acids was truncated from VP24 and several mutants were prepared to increase the proportion of selenomethionine (SeMet) residues for subsequent structural determination using the SAD method. Truncated VP24, its mutants and the corresponding SeMet-labelled proteins were purified, and the native and SeMet proteins were crystallized by the hanging-drop vapour-diffusion method. Crystals of VP24 were obtained using a reservoir consisting of 0.1 M Tris-HCl pH 8.5, 2.75 M ammonium acetate with a drop volume ratio of two parts protein solution to one part reservoir solution. Notably, ATP was added as a critical additive to the drop with a final concentration of 10 mM. Crystals of SeMet-labelled VP24 mutant diffracted to 3.0 Å resolution and those of the native diffracted to 2.4 Å resolution; the crystals belonged to space group I213, with unit-cell parameters a = b = c = 140 Å.

  19. Crayfish hematopoietic tissue cells but not hemocytes are permissive for white spot syndrome virus replication.

    PubMed

    Wu, Junjun; Li, Fang; Huang, Jiajun; Xu, Limei; Yang, Feng

    2015-03-01

    Hemocytes are the major immune cells of crustaceans which are believed to be essential for the pathogenesis of white spot syndrome virus (WSSV) infection. Crayfish hemocytes and hematopoietic tissue (HPT) cells have been found to be susceptible to WSSV infection, but the procedure of WSSV infection to both cell types has not yet been carefully investigated. In this study, we analyzed the infection and proliferation of WSSV in crayfish hemocytes as well as HPT cells in detail through transmission electronic microscopy (TEM). The results showed that WSSV could enter both hemocytes and HPT cells through endocytosis, but the production of progeny virus was only achieved in HPT cells. Further investigation demonstrated that although WSSV could transcribe its genes in both cell types, viral genome replication and structural protein expression were unsuccessful in hemocytes, which may be responsible for the failure of progeny production. Therefore, we propose that both hemocytes and HPT cells are susceptible to WSSV infection but only HPT cells are permissive to WSSV replication. These findings will extend our knowledge of the interaction between WSSV and the host immune system.

  20. Comparative genomic analysis of three white spot syndrome virus isolates of different virulence.

    PubMed

    Li, Fang; Gao, Meiling; Xu, Limei; Yang, Feng

    2017-04-01

    Three white spot syndrome virus (WSSV) isolates of different virulence were identified in our previous study, the high-virulent strain WSSV-CN01, the moderate-virulent strain WSSV-CN02 and the low-virulent strain WSSV-CN03. In this study, the genomes of these three WSSV isolates were sequenced, annotated and compared. The genome sizes for WSSV-CN01, WSSV-CN02, and WSSV-CN03 are 309,286, 294,261, and 284,148 bp, bearing 177, 164, and 154 putative protein-coding genes, respectively. The genomic variations including insertions, deletions, and substitutions were investigated. Thirty four genes show >20% variation in their sequences in WSSV-CN02 or WSSV-CN03, in comparison with WSSV-CN01, including six envelope protein genes (wsv237/vp41A, wsv238/vp52A, wsv338/vp62, wsv339/vp39, wsv077/vp36A, and wsv242/vp41B), and two immediate-early genes (wsv108 and wsv178). The genomic variations among WSSV isolates of different virulence, especially those in the coding regions, certainly provide new insight into the understanding of the molecular basis of WSSV pathogenesis.

  1. White spot syndrome virus VP12 interacts with adenine nucleotide translocase of Litopenaeus vannamei.

    PubMed

    Ma, Fang-fang; Chou, Zhi-guang; Liu, Qing-hui; Guan, Guangkuo; Li, Chen; Huang, Jie

    2014-05-01

    White spot syndrome virus VP12 contains cell attachment motif RGD which is considered to be critical for host cell binding. Until now, the function of this protein remains undefined. In this study, we explored the interaction of VP12 with host cells. A new shrimp protein (adenine nucleotide translocase of Litopenaeus vannamei, LvANT) is selected by far-western overlay assay. Tissue distribution of adenine nucleotide translocase mRNA showed that it was commonly spread in all the tissues detected. Cellular localization of LvANT in shrimp hemocytes showed that it was primarily located in the cytoplasm of hemocytes and colocalized with mitochondria. ELISA and far-western blot assay confirmed that VP12 interacted with LvANT. In vivo neutralization assay showed that anti-LvANT antibody can significantly reduce the mortality of shrimp challenged by WSSV at 48h post-treatment. Our results collectively showed that VP12 is involved in host cell binding via interaction with adenine nucleotide translocase.

  2. The tidepool shrimp, Palaemon ritteri Holmes, constitutes a novel host to the white spot syndrome virus.

    PubMed

    Sánchez-Paz, A; Terán-Díaz, B; Enríquez-Espinoza, T; Encinas-Garcia, T; Vázquez-Sánchez, I; Mendoza-Cano, F

    2015-07-01

    The white spot syndrome virus (WSSV) is a lethal and contagious pathogen for penaeid shrimp and a growing number of other crustacean species. To date, there are no effective prophylactic or therapeutic treatments commercially available to interfere with the occurrence and spread of the disease. In addition, the significance of alternative vectors on the dispersal of this disease has been largely ignored and therefore the ecological dynamics of the WSSV is still poorly understood and difficult to ascertain. Thus, an important issue that should be considered in sanitary programmes and management strategies is the identification of species susceptible to infection by WSSV. The results obtained provide the first direct evidence of ongoing WSSV replication in experimentally infected specimens of the tidepool shrimp Palaemon ritteri. Viral replication was detected using a validated set of primers for the amplification by RT-PCR of a 141 bp fragment of the transcript encoding the viral protein VP28. It is therefore conceivable that this shrimp may play a significant role in the dispersal of WSSV.

  3. White spot syndrome virus infection: Threat to crustacean biodiversity in Vembanad Lake, India.

    PubMed

    Joseph, Toms C; James, Roswin; Rajan, L Anbu; Surendran, P K; Lalitha, K V

    2015-09-01

    The Vembanad Lake located on the south-west coast of India, an ecological hotspot is the nursing ground of many economically important crustaceans. The prevalence of white spot syndrome virus (WSSV) among crustaceans from farmed, estuarine and marine environments surrounding the Vembanad Lake, India was detected using PCR. A total of 308 samples from aquaculture ponds consisting of six species of crustaceans collected from five different farms were tested for the presence of WSSV. Of these, 67% were found to carry the virus. A total of 258 samples of crustaceans from the Cochin backwater system that forms a part of the Vembanad lake viz., Metapenaeus dobsoni, Metapenaeus monoceros, Penaeus monodon and Penaeus indicus were found to contain WSSV in 62% of the samples. Fifteen species of crustaceans caught from the seas off Cochin were also screened for the presence of WSSV. Out of these, twelve species had WSSV incidence levels ranging from 6-23%. WSSV was not detected from three species of deep sea crustaceans tested. The black tiger shrimp, Penaeus monodon had the highest incidence of WSSV among the species screened in farmed, estuarine and marine environments.

  4. Purification of white spot syndrome virus by iodixanol density gradient centrifugation.

    PubMed

    Dantas-Lima, J J; Corteel, M; Cornelissen, M; Bossier, P; Sorgeloos, P; Nauwynck, H J

    2013-10-01

    Up to now, only a few brief procedures for purifying white spot syndrome virus (WSSV) have been described. They were mainly based on sucrose, NaBr and CsCl density gradient centrifugation. This work describes for the first time the purification of WSSV through iodixanol density gradients, using virus isolated from infected tissues and haemolymph of Penaeus vannamei (Boone). The purification from tissues included a concentration step by centrifugation (2.5 h at 60,000 g) onto a 50% iodixanol cushion and a purification step by centrifugation (3 h at 80,000 g) through a discontinuous iodixanol gradient (phosphate-buffered saline, 5%, 10%, 15% and 20%). The purification from infected haemolymph enclosed a dialysis step with a membrane of 1,000 kDa (18 h) and a purification step through the earlier iodixanol gradient. The gradients were collected in fractions and analysed. The number of particles, infectivity titre (in vivo), total protein and viral protein content were evaluated. The purification from infected tissues gave WSSV suspensions with a very high infectivity and an acceptable purity, while virus purified from haemolymph had a high infectivity and a very high purity. Additionally, it was observed that WSSV has an unusually low buoyant density and that it is very sensitive to high external pressures.

  5. Evidence for cell apoptosis suppressing white spot syndrome virus replication in Procambarus clarkii at high temperature.

    PubMed

    Wu, Xiao-Guo; Xiong, Hai-Tao; Wang, Yi-Zhen; Du, Hua-Hua

    2012-12-03

    In shrimp, higher water temperatures (~32°C) can suppress the ability of white spot syndrome virus (WSSV) to replicate and cause mortality, but the mechanisms remain unclear. To investigate whether cell apoptosis might be involved, a Tdt-mediated dUTP nick-end label (TUNEL) method was used to assess levels of chromosomal DNA fragmentation in hepatopancreas and gill cells of Procambarus clarkii crayfish infected with WSSV and maintained at either 32 ± 1°C or 24 ± 1°C. Based on relative cell numbers with yellow-green colored TUNEL-positive nuclei, the apoptotic index was elevated in WSSV-infected crayfish maintained at 32°C. In gill tissue sections examined by transmission electron microscope, cells with nuclei displaying apoptotic bodies or marginated, condensed and fragmented chromatin without concurrent cell cytoplasm damage were also more prevalent. Flow cytometry sorting of annexin-stained cells showed apoptosis to be most prevalent in granular haemocytes, and assays for caspase-3 activity showed it to be most elevated in hepatopancreas tissue. Despite these indicators of cell apoptosis but consistent with WSSV replication being restricted at elevated temperatures, no increases in transcription of the viral anti-apoptosis genes ORF390 and ORF222 were detected by RT-PCR in shrimp maintained at 32°C, possibly due to the elevated levels of cellular apoptosis.

  6. Screening white spot syndrome virus (WSSV)-resistant molecular markers from Fenneropenaeus chinensis

    NASA Astrophysics Data System (ADS)

    Wu, Yingying; Meng, Xianhong; Kong, Jie; Luan, Sheng; Luo, Kun; Wang, Qingyin; Zheng, Yongyun

    2017-02-01

    White spot syndrome virus (WSSV)-resistant molecular markers were screened from the selectively bred new variety `Huanghai No. 2' of Fenneropenaeus chinensis using unlabeled-probe high-resolution melting (HRM) technique. After the artificial infection with WSSV, the first 96 dead shrimps and the last 96 surviving shrimps were collected, representing WSSV-susceptible and -resistant populations, respectively. The genotypes at well-developed 39 single nucleotide polymorphisms (SNPs) loci were obtained. As revealed in the Chi-square test, 3 SNPs, genotype A/A of contig C364-89AT, genotype A/A of C2635-527CA and genotype C/T of contig C12355-592CT, were positively correlated with disease-resistance traits. Other 2 SNPs, genotype G/G of contig C283-145AG and genotype C/C of contig C12355-592CT, were negatively correlated. Moreover, analysis with BlastX program for disease-resistant SNPs indicated that 3 contigs, Contig283, Contig364 and Contig12355, matched to the functional genes of effector caspase of Penaeus monodon, peptide transporter family 1-like protein, and 40S ribosomal protein S2 of Perca flavescens with high sequence similarity. The results will be helpful to provide theoretical and technical supports for molecular marker-assisted selective breeding of F. chinensis.

  7. Recent insights into host-pathogen interaction in white spot syndrome virus infected penaeid shrimp.

    PubMed

    Shekhar, M S; Ponniah, A G

    2015-07-01

    Viral disease outbreaks are a major concern impeding the development of the shrimp aquaculture industry. The viral disease due to white spot syndrome virus (WSSV) observed in early 1990s still continues unabated affecting the shrimp farms and cause huge economic loss to the shrimp aquaculture industry. In the absence of effective therapeutics to control WSSV, it is important to understand viral pathogenesis and shrimp response to WSSV at the molecular level. Identification and molecular characterization of WSSV proteins and receptors may facilitate in designing and development of novel therapeutics and antiviral drugs that may inhibit viral replication. Investigations into host-pathogen interactions might give new insights to viral infectivity, tissue tropism and defence mechanism elicited in response to WSSV infection. However, due to the limited information on WSSV gene function and host immune response, the signalling pathways which are associated in shrimp pathogen interaction have also not been elucidated completely. In the present review, the focus is on those shrimp proteins and receptors that are potentially involved in virus infection or in the defence mechanism against WSSV. In addition, the major signalling pathways involved in the innate immune response and the role of apoptosis in host-pathogen interaction is discussed.

  8. Transmission of white spot syndrome virus (WSSV) from Dendronereis spp. (Peters) (Nereididae) to penaeid shrimp.

    PubMed

    Haryadi, D; Verreth, J A J; Verdegem, M C J; Vlak, J M

    2015-05-01

    Dendronereis spp. (Peters) (Nereididae) is a common polychaete in shrimp ponds built on intertidal land and is natural food for shrimp in traditionally managed ponds in Indonesia. White spot syndrome virus (WSSV), an important viral pathogen of the shrimp, can replicate in this polychaete (Desrina et al. 2013); therefore, it is a potential propagative vector for virus transmission. The major aim of this study was to determine whether WSSV can be transmitted from naturally infected Dendronereis spp. to specific pathogen-free (SPF) Pacific white shrimp Litopenaeus vannamei (Boone) through feeding. WSSV was detected in naturally infected Dendronereis spp. and Penaeus monodon Fabricius from a traditional shrimp pond, and the positive animals were used in the current experiment. WSSV-infected Dendronereis spp. and P. monodon in a pond had a point prevalence of 90% and 80%, respectively, as measured by PCR. WSSV was detected in the head, gills, blood and mid-body of Dendronereis spp. WSSV from naturally infected Dendronereis spp was transmitted to SPF L. vannamei and subsequently from this shrimp to new naïve-SPF L. vannamei to cause transient infection. Our findings support the contention that Dendronereis spp, upon feeding, can be a source of WSSV infection of shrimp in ponds.

  9. Construction and application of a protein interaction map for white spot syndrome virus (WSSV).

    PubMed

    Sangsuriya, Pakkakul; Huang, Jiun-Yan; Chu, Yu-Fei; Phiwsaiya, Kornsunee; Leekitcharoenphon, Pimlapas; Meemetta, Watcharachai; Senapin, Saengchan; Huang, Wei-Pang; Withyachumnarnkul, Boonsirm; Flegel, Timothy W; Lo, Chu-Fang

    2014-01-01

    White spot syndrome virus (WSSV) is currently the most serious global threat for cultured shrimp production. Although its large, double-stranded DNA genome has been completely characterized, most putative protein functions remain obscure. To provide more informative knowledge about this virus, a proteomic-scale network of WSSV-WSSV protein interactions was carried out using a comprehensive yeast two-hybrid analysis. An array of yeast transformants containing each WSSV open reading frame fused with GAL4 DNA binding domain and GAL4 activation domain was constructed yielding 187 bait and 182 prey constructs, respectively. On screening of ∼28,000 pairwise combinations, 710 interactions were obtained from 143 baits. An independent coimmunoprecipitation assay (co-IP) was performed to validate the selected protein interaction pairs identified from the yeast two-hybrid approach. The program Cytoscape was employed to create a WSSV protein-protein interaction (PPI) network. The topology of the WSSV PPI network was based on the Barabási-Albert model and consisted of a scale-free network that resembled other established viral protein interaction networks. Using the RNA interference approach, knocking down either of two candidate hub proteins gave shrimp more protection against WSSV than knocking down a nonhub gene. The WSSV protein interaction map established in this study provides novel guidance for further studies on shrimp viral pathogenesis, host-viral protein interaction and potential targets for therapeutic and preventative antiviral strategies in shrimp aquaculture.

  10. Electroless-plated gold films for sensitive surface plasmon resonance detection of white spot syndrome virus.

    PubMed

    Lei, Yun; Chen, Hongyu; Dai, Heping; Zeng, Zhaorui; Lin, Yi; Zhou, Feimeng; Pang, Daiwen

    2008-02-28

    The paper describes the rapid and label-free detection of the white spot syndrome virus (WSSV) using a surface plasmon resonance (SPR) device based on gold films prepared by electroless plating. The plating condition for obtaining films suitable for SPR measurements was optimized. Gold nanoparticles adsorbed on glass slides were characterized by transmission electron microscopy (TEM). Detection of the WSSV was performed through the binding between WSSV in solution and the anti-WSSV single chain variable fragment (scFv antibody) preimmobilized onto the sensor surface. Morphologies of the as-prepared gold films, gold films modified with self-assembled alkanethiol monolayers, and films covered with antibody were examined using an atomic force microscope (AFM). To demonstrate the viability of the method for real sample analysis, WSSV of different concentrations present in a shrimp hemolymph matrix was determined upon optimizing the surface density of the antibody molecules. The SPR device based on the electroless-plated gold films is capable of detecting concentration of WSSV as low as 2.5 ng/mL in 2% shrimp hemolymph, which is one to two orders of magnitude lower than the level measurable by enzyme-linked immunosorbant assays.

  11. Rapid diagnosis of vibriosis and white spot syndrome (WSS) in the culture of shrimp, Penaeus monodon in Philippines.

    PubMed

    Caipang, Christopher Marlowe A; Aguana, Mary Paz N

    2010-10-01

    Viral and bacterial pathogens have raised serious concerns in the sustainability of the shrimp culture industry in the Philippines. Heavy mortality associated with luminous vibriosis and white spot syndrome virus (WSSV) infection has been the major problem besetting the industry. Using published PCR protocols for the diagnosis of vibriosis and white spot syndrome virus (WSSV) disease in shrimp, we optimized these assays that could be suited to the shrimp aquaculture setting in the Philippines. Genomic DNAs of Vibrio spp. that exhibited luminescence as well as those that grew on thiosulfate citrate bile salt sucrose agar (TCBS) were used for the PCR amplification of the ribonuclease P (RNase P) gene. There was differential amplification of the RNase P gene based on the phenotypic characters of the Vibrio spp. Similar results were also obtained using direct colony PCR of the bacterial colonies. White spot syndrome virus was also detected in the infected shrimp and there were differences in the detection frequency in relation to the tissues used for PCR amplification. Duplex PCR was also optimized that could be used for simultaneous detection of these pathogens in shrimp.

  12. A model for apoptotic interaction between white spot syndrome virus and shrimp.

    PubMed

    Leu, Jiann-Horng; Lin, Shin-Jen; Huang, Jiun-Yan; Chen, Tsan-Chi; Lo, Chu-Fang

    2013-04-01

    White spot syndrome virus (WSSV) is an enveloped, large dsDNA virus that mainly infects penaeid shrimp, causing serious damage to the shrimp aquaculture industry. Like other animal viruses, WSSV infection induces apoptosis. Although this occurs even in by-stander cells that are free of WSSV virions, apoptosis is generally regarded as a kind of antiviral immune response. To counter this response, WSSV has evolved several different strategies. From the presently available literature, we construct a model of how the host and virus both attempt to regulate apoptosis to their respective advantage. The basic sequence of events is as follows: first, when a WSSV infection occurs, cellular sensors detect the invading virus, and activate signaling pathways that lead to (1) the expression of pro-apoptosis proteins, including PmCasp (an effecter caspase), MjCaspase (an initiator caspase) and voltage-dependent anion channel (VDAC); and (2) mitochondrial changes, including the induction of mitochondrial membrane permeabilization and increased oxidative stress. These events initiate the apoptosis program. Meanwhile, WSSV begins to express its genes, including two anti-apoptosis proteins: AAP-1, which is a direct caspase inhibitor, and WSV222, which is an E3 ubiquitin ligase that blocks apoptosis through the ubiquitin-mediated degradation of shrimp TSL protein (an apoptosis inducer). WSSV also induces the expression of a shrimp anti-apoptosis protein, Pm-fortilin, which can act on Bax to inhibit mitochondria-triggered apoptosis. This is a life and death struggle because the virus needs to prevent apoptosis in order to replicate. If WSSV succeeds in replicating in sufficient numbers, this will result in the death of the infected penaeid shrimp host.

  13. Scavenger Receptor C Mediates Phagocytosis of White Spot Syndrome Virus and Restricts Virus Proliferation in Shrimp.

    PubMed

    Yang, Ming-Chong; Shi, Xiu-Zhen; Yang, Hui-Ting; Sun, Jie-Jie; Xu, Ling; Wang, Xian-Wei; Zhao, Xiao-Fan; Wang, Jin-Xing

    2016-12-01

    Scavenger receptors are an important class of pattern recognition receptors that play several important roles in host defense against pathogens. The class C scavenger receptors (SRCs) have only been identified in a few invertebrates, and their role in the immune response against viruses is seldom studied. In this study, we firstly identified an SRC from kuruma shrimp, Marsupenaeus japonicus, designated MjSRC, which was significantly upregulated after white spot syndrome virus (WSSV) challenge at the mRNA and protein levels in hemocytes. The quantity of WSSV increased in shrimp after knockdown of MjSRC, compared with the controls. Furthermore, overexpression of MjSRC led to enhanced WSSV elimination via phagocytosis by hemocytes. Pull-down and co-immunoprecipitation assays demonstrated the interaction between MjSRC and the WSSV envelope protein. Electron microscopy observation indicated that the colloidal gold-labeled extracellular domain of MjSRC was located on the outer surface of WSSV. MjSRC formed a trimer and was internalized into the cytoplasm after WSSV challenge, and the internalization was strongly inhibited after knockdown of Mjβ-arrestin2. Further studies found that Mjβ-arrestin2 interacted with the intracellular domain of MjSRC and induced the internalization of WSSV in a clathrin-dependent manner. WSSV were co-localized with lysosomes in hemocytes and the WSSV quantity in shrimp increased after injection of lysosome inhibitor, chloroquine. Collectively, this study demonstrated that MjSRC recognized WSSV via its extracellular domain and invoked hemocyte phagocytosis to restrict WSSV systemic infection. This is the first study to report an SRC as a pattern recognition receptor promoting phagocytosis of a virus.

  14. Shrimp MyD88 responsive to bacteria and white spot syndrome virus.

    PubMed

    Wen, Rong; Li, Fuhua; Sun, Zheng; Li, Shihao; Xiang, Jianhai

    2013-02-01

    The myeloid differentiation factor 88 (MyD88) is an important adapter protein which links members of the toll-like receptor (TLR) to the downstream components to activate related signaling pathways. In the present study, a MyD88 homolog (FcMyD88) was cloned from penaeid shrimp Fenneropenaeus chinensis. The ORF of FcMyD88 consisted of 1434 bp encoding a polypeptide of 477 amino acids which contains a death domain (DD) and a typical TLR and interleukin-1 receptor (IL-1R)-related (TIR) domain. Homology analysis revealed that the predicted amino acid (aa) sequence of FcMyD88 shared high similarities with a variety of previously reported MyD88s. The time-dependent expression patterns of FcMyD88 in cephalothoraxes of shrimp injected with Vibrio anguillarum (Gram-negative bacteria, G(-)), Micrococcus lysodeikticu (Gram-positive bacteria, G(+)) and white syndrome spot virus (WSSV) were analyzed at transcription and protein level by real-time PCR and western blotting, respectively. The expression level of FcMyD88 mRNA was significantly up-regulated at one hour (h), 12 h and 24 h after stimulation with both V. anguillarum and M. lysodeikticu. The expression level of FcMyD88 protein was 2-fold up-regulated at 12 h post injection (hpi) of inactivated V. anguillarum while it didn't change after M. lysodeikticu injection during this period. After WSSV injection, the expression level of FcMyD88 mRNA remained relatively constant, while the FcMyD88 protein was significantly up-regulated at 12 and 24 hpi. These results suggested that the MyD88-dependent signaling pathway could be involved in the defense of both bacteria and WSSV infection.

  15. Evolutionary Trajectory of White Spot Syndrome Virus (WSSV) Genome Shrinkage during Spread in Asia

    PubMed Central

    Hemerik, Lia; Vlak, Just M.

    2010-01-01

    Background White spot syndrome virus (WSSV) is the sole member of the novel Nimaviridae family, and the source of major economic problems in shrimp aquaculture. WSSV appears to have rapidly spread worldwide after the first reported outbreak in the early 1990s. Genomic deletions of various sizes occur at two loci in the WSSV genome, the ORF14/15 and ORF23/24 variable regions, and these have been used as molecular markers to study patterns of viral spread over space and time. We describe the dynamics underlying the process of WSSV genome shrinkage using empirical data and a simple mathematical model. Methodology/Principal Findings We genotyped new WSSV isolates from five Asian countries, and analyzed this information together with published data. Genome size appears to stabilize over time, and deletion size in the ORF23/24 variable region was significantly related to the time of the first WSSV outbreak in a particular country. Parameter estimates derived from fitting a simple mathematical model of genome shrinkage to the data support a geometric progression (k<1) of the genomic deletions, with k = 0.371±0.150. Conclusions/Significance The data suggest that the rate of genome shrinkage decreases over time before attenuating. Bioassay data provided support for a link between genome size and WSSV fitness in an aquaculture setting. Differences in genomic deletions between geographic WSSV isolates suggest that WSSV spread did not follow a smooth pattern of geographic radiation, suggesting spread of WSSV over long distances by commercial activities. We discuss two hypotheses for genome shrinkage, an adaptive and a neutral one. We argue in favor of the adaptive hypothesis, given that there is support for a link between WSSV genome size and fitness. PMID:20976239

  16. Tangential flow ultrafiltration for detection of white spot syndrome virus (WSSV) in shrimp pond water.

    PubMed

    Alavandi, S V; Ananda Bharathi, R; Satheesh Kumar, S; Dineshkumar, N; Saravanakumar, C; Joseph Sahaya Rajan, J

    2015-06-15

    Water represents the most important component in the white spot syndrome virus (WSSV) transmission pathway in aquaculture, yet there is very little information. Detection of viruses in water is a challenge, since their counts will often be too low to be detected by available methods such as polymerase chain reaction (PCR). In order to overcome this difficulty, viruses in water have to be concentrated from large volumes of water prior to detection. In this study, a total of 19 water samples from aquaculture ecosystem comprising 3 creeks, 10 shrimp culture ponds, 3 shrimp broodstock tanks and 2 larval rearing tanks of shrimp hatcheries and a sample from a hatchery effluent treatment tank were subjected to concentration of viruses by ultrafiltration (UF) using tangential flow filtration (TFF). Twenty to 100l of water from these sources was concentrated to a final volume of 100mL (200-1000 fold). The efficiency of recovery of WSSV by TFF ranged from 7.5 to 89.61%. WSSV could be successfully detected by PCR in the viral concentrates obtained from water samples of three shrimp culture ponds, one each of the shrimp broodstock tank, larval rearing tank, and the shrimp hatchery effluent treatment tank with WSSV copy numbers ranging from 6 to 157mL(-1) by quantitative real time PCR. The ultrafiltration virus concentration technique enables efficient detection of shrimp viral pathogens in water from aquaculture facilities. It could be used as an important tool to understand the efficacy of biosecurity protocols adopted in the aquaculture facility and to carry out epidemiological investigations of aquatic viral pathogens.

  17. Evolution of specific immunity in shrimp - a vaccination perspective against white spot syndrome virus.

    PubMed

    Syed Musthaq, Syed Khader; Kwang, Jimmy

    2014-10-01

    Invertebrates lack true adaptive immunity and it solely depends on the primitive immunity called innate immunity. However, various innate immune molecules and mechanisms are identified in shrimp that plays potential role against invading bacterial, fungal and viral pathogens. Perceiving the shrimp innate immune mechanisms will contribute in developing effective vaccine strategies against major shrimp pathogens. Hence this review intends to explore the innate immune molecules of shrimp with suitable experimental evidences together with the evolution of "specific immune priming" of invertebrates. In addition, we have emphasized on the development of an effective vaccine strategy against major shrimp pathogen, white spot syndrome virus (WSSV). The baculovirus displayed rVP28 (Bac-VP28), a major envelope protein of WSSV was utilized to study its vaccine efficacy by oral route. A significant advantage of this baculovirus expression cassette is the use of WSSV-immediate early 1 (ie1) promoter that derived the abundant expression of rVP28 protein at the early stage of the infection in insect cell. The orally vaccinated shrimp with Bac-VP28 transduced successfully in the shrimp cells as well as provided highest survival rate. In support to our vaccine efficacy we analysed Pattern Recognition Proteins (PRPs) β-1,3 glucan lipopolysaccharides (LGBP) and STAT gene profiles in the experimental shrimp. Indeed, the vaccination of shrimp with Bac-VP28 demonstrated some degree of specificity with enhanced survival rate when compared to control vaccination with Bac-wt. Hence it is presumed that the concept of "specific immune priming" in relevant to shrimp immunity is possible but may not be common to all shrimp pathogens.

  18. Expression, Purification, Crystallization of Two Major Envelope Proteins from White Spot Syndrome Virus

    SciTech Connect

    Tang,X.; Hew, C.

    2007-01-01

    White spot syndrome virus (WSSV) is a major virulent pathogen known to infect penaeid shrimp and other crustaceans. VP26 and VP28, two major envelope proteins from WSSV, have been identified and overexpressed in Escherichia coli. In order to facilitate purification and crystallization, predicted N-terminal transmembrane regions of approximately 35 amino acids have been truncated from both VP26 and VP28. Truncated VP26 and VP28 and their corresponding SeMet-labelled proteins were purified and the SeMet proteins were crystallized by the hanging-drop vapor-diffusion method. Crystals of SeMet-labelled VP26 were obtained using a reservoir consisting of 0.1 M citric acid pH 3.5, 3.0 M sodium chloride and 1%(w/v) polyethylene glycol 3350, whereas SeMet VP28 was crystallized using a reservoir solution consisting of 25% polyethylene glycol 8000, 0.2 M calcium acetate, 0.1 M Na HEPES pH 7.5 and 1.5%(w/v) 1,2,3-heptanetriol. Crystals of SeMet-labelled VP26 diffract to 2.2 {angstrom} resolution and belong to space group R32, with unit-cell parameters a = b = 73.92, c = 199.31 {angstrom}. SeMet-labelled VP28 crystallizes in space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 105.33, b = 106.71, c = 200.37 {angstrom}, and diffracts to 2.0 {angstrom} resolution.

  19. Shrimp miRNAs regulate innate immune response against white spot syndrome virus infection.

    PubMed

    Kaewkascholkul, Napol; Somboonviwat, Kulwadee; Asakawa, Shuichi; Hirono, Ikuo; Tassanakajon, Anchalee; Somboonwiwat, Kunlaya

    2016-07-01

    MicroRNAs are short noncoding RNAs of RNA interference pathways that regulate gene expression through partial complementary base-pairing to target mRNAs. In this study, miRNAs that are expressed in white spot syndrome virus (WSSV)-infected Penaeus monodon, were identified using next generation sequencing. Forty-six miRNA homologs were identified from WSSV-infected shrimp hemocyte. Stem-loop real-time RT-PCR analysis showed that 11 out of 16 selected miRNAs were differentially expressed upon WSSV infection. Of those, pmo-miR-315 and pmo-miR-750 were highly responsive miRNAs. miRNA target prediction revealed that the miRNAs were targeted at 5'UTR, ORF, and 3'UTR of several immune-related genes such as genes encoding antimicrobial peptides, signaling transduction proteins, heat shock proteins, oxidative stress proteins, proteinases or proteinase inhibitors, proteins in blood clotting system, apoptosis-related proteins, proteins in prophenoloxidase system, pattern recognition proteins and other immune molecules. The highly conserved miRNA homolog, pmo-bantam, was characterized for its function in shrimp. The pmo-bantam was predicted to target the 3'UTR of Kunitz-type serine protease inhibitor (KuSPI). Binding of pmo-bantam to the target sequence of KuSPI gene was analyzed by luciferase reporter assay. Correlation of pmo-bantam and KuSPI expression was observed in lymphoid organ of WSSV-infected shrimp. These results implied that miRNAs might play roles as immune gene regulators in shrimp antiviral response.

  20. Reprint of "evolution of specific immunity in shrimp - a vaccination perspective against white spot syndrome virus".

    PubMed

    Syed Musthaq, Syed Khader; Kwang, Jimmy

    2015-02-01

    Invertebrates lack true adaptive immunity and it solely depends on the primitive immunity called innate immunity. However, various innate immune molecules and mechanisms are identified in shrimp that plays potential role against invading bacterial, fungal and viral pathogens. Perceiving the shrimp innate immune mechanisms will contribute in developing effective vaccine strategies against major shrimp pathogens. Hence this review intends to explore the innate immune molecules of shrimp with suitable experimental evidences together with the evolution of "specific immune priming" of invertebrates. In addition, we have emphasized on the development of an effective vaccine strategy against major shrimp pathogen, white spot syndrome virus (WSSV). The baculovirus displayed rVP28 (Bac-VP28), a major envelope protein of WSSV was utilized to study its vaccine efficacy by oral route. A significant advantage of this baculovirus expression cassette is the use of WSSV-immediate early 1 (ie1) promoter that derived the abundant expression of rVP28 protein at the early stage of the infection in insect cell. The orally vaccinated shrimp with Bac-VP28 transduced successfully in the shrimp cells as well as provided highest survival rate. In support to our vaccine efficacy we analysed Pattern Recognition Proteins (PRPs) β-1,3 glucan lipopolysaccharides (LGBP) and STAT gene profiles in the experimental shrimp. Indeed, the vaccination of shrimp with Bac-VP28 demonstrated some degree of specificity with enhanced survival rate when compared to control vaccination with Bac-wt. Hence it is presumed that the concept of "specific immune priming" in relevant to shrimp immunity is possible but may not be common to all shrimp pathogens.

  1. VP24 Is a Chitin-Binding Protein Involved in White Spot Syndrome Virus Infection

    PubMed Central

    Li, Zaipeng; Han, Yali; Xu, Limei

    2015-01-01

    ABSTRACT Oral ingestion is the major route of infection for the white spot syndrome virus (WSSV). However, the mechanism by which virus particles in the digestive tract invade host cells is unknown. In the present study, we demonstrate that WSSV virions can bind to chitin through one of the major envelope proteins (VP24). Mutagenesis analysis indicated that amino acids (aa) 186 to 200 in the C terminus of VP24 were required for chitin binding. Moreover, the P-VP24186–200 peptide derived from the VP24 chitin binding region significantly inhibited the VP24-chitin interaction and the WSSV-chitin interaction, implying that VP24 participates in WSSV binding to chitin. Oral inoculation experiments showed that P-VP24186–200 treatment reduced the number of virus particles remaining in the digestive tract during the early stage of infection and greatly hindered WSSV proliferation in shrimp. These data indicate that binding of WSSV to chitin through the viral envelope protein VP24 is essential for WSSV per os infection and provide new ideas for preventing WSSV infection in shrimp farms. IMPORTANCE In this study, we show that WSSV can bind to chitin through the envelope protein VP24. The chitin-binding domain of VP24 maps to amino acids 186 to 200 in the C terminus. Binding of WSSV to chitin through the viral envelope protein VP24 is essential for WSSV per os infection. These findings not only extend our knowledge of WSSV infection but also provide new insights into strategies to prevent WSSV infection in shrimp farms. PMID:26512091

  2. Scavenger Receptor C Mediates Phagocytosis of White Spot Syndrome Virus and Restricts Virus Proliferation in Shrimp

    PubMed Central

    Yang, Ming-Chong; Shi, Xiu-Zhen; Yang, Hui-Ting; Sun, Jie-Jie; Xu, Ling; Wang, Xian-Wei; Zhao, Xiao-Fan

    2016-01-01

    Scavenger receptors are an important class of pattern recognition receptors that play several important roles in host defense against pathogens. The class C scavenger receptors (SRCs) have only been identified in a few invertebrates, and their role in the immune response against viruses is seldom studied. In this study, we firstly identified an SRC from kuruma shrimp, Marsupenaeus japonicus, designated MjSRC, which was significantly upregulated after white spot syndrome virus (WSSV) challenge at the mRNA and protein levels in hemocytes. The quantity of WSSV increased in shrimp after knockdown of MjSRC, compared with the controls. Furthermore, overexpression of MjSRC led to enhanced WSSV elimination via phagocytosis by hemocytes. Pull-down and co-immunoprecipitation assays demonstrated the interaction between MjSRC and the WSSV envelope protein. Electron microscopy observation indicated that the colloidal gold-labeled extracellular domain of MjSRC was located on the outer surface of WSSV. MjSRC formed a trimer and was internalized into the cytoplasm after WSSV challenge, and the internalization was strongly inhibited after knockdown of Mjβ-arrestin2. Further studies found that Mjβ-arrestin2 interacted with the intracellular domain of MjSRC and induced the internalization of WSSV in a clathrin-dependent manner. WSSV were co-localized with lysosomes in hemocytes and the WSSV quantity in shrimp increased after injection of lysosome inhibitor, chloroquine. Collectively, this study demonstrated that MjSRC recognized WSSV via its extracellular domain and invoked hemocyte phagocytosis to restrict WSSV systemic infection. This is the first study to report an SRC as a pattern recognition receptor promoting phagocytosis of a virus. PMID:28027319

  3. Litopenaeus vannamei clathrin coat AP17 involved in white spot syndrome virus infection.

    PubMed

    Wang, Xiu-Fang; Liu, Qing-Hui; Wu, Yin; Huang, Jie

    2016-05-01

    White spot syndrome virus (WSSV) is the main pathogen of shrimp culture, and has brought great losses of the shrimp aquaculture industry every year since it has been found. However, the specific mechanism of the virus into the cell is not very clear. Recent research suggests that clathrin-mediated endocytosis is involved in WSSV infection. By sequence analysis, clathrin coat AP17 is an σ subunit of AP-2 complex which is involved in clathrin-mediated endocytosis. To obtain the full-length sequence of Clathrin coat AP17 of Litopenaeus vannamei (LvCCAP17), the rapid amplification of cDNA ends (RACE) was performed to get the sequence of 3'and 5' end and splicing by DNAMAN. The full-length sequence of LvCCAP17 is 842 bp and expected to encoding 142 amino acids, and the amino acid sequence was analyzed by online software. The mRNA expression of LvCCAP17 in different tissues was carried out with quantitative real-time PCR and the LvCCAP17 was detected in all tested tissues of Litopenaeus vannamei. The transcriptional expression level of LvCCAP17 in epithelium and hepatopancreas was significantly up-regulated after WSSV infection. Far-Western blotting and ELISA assay showed that LvCCAP17 interacted with rVP26 and rVP37. Silencing of LvCCAP17 gene by double-strand RNA (dsRNA) interference significantly delay of cumulative mortality rate in WSSV infected shrimp and reduced the expression level of immediate early gene 1(ie1) and vp28. These results indicated that clathrin-meated endocytosis is responsible for WSSV infection.

  4. McCune Albright syndrome - association of fibrous dysplasia, café-au-lait skin spots and hyperthyroidism - case report.

    PubMed

    Raus, Iulian; Coroiu, Roxana Elena

    2016-01-01

    McCune-Albright syndrome is a rare sporadic disease characterized by bone fibrous dysplasia, café-au-lait skin spots and a variable association of hyperfunctional endocrine disorders. Fibrous dysplasia (FD), which can involve the craniofacial, axial, and appendicular skeleton, may range from an isolated, asymptomatic monostotic lesion to a severe disabling polyostotic disease involving the entire skeleton. A twenty-five-year old male patient presented to our clinic with recently developed heart palpitations. He had also been feeling pain in the right femur since he was younger, without any trauma history, leading to difficulties of ambulation and limping occasionally. His physical examination revealed café-au-lait spots with irregular borders and right testicular agenesis. Laboratory findings identified hyperthyroidism with hyperparathyroidism. Radiographs of the pelvis revealed multiple lytic lesions of the right femur and magnetic resonance imaging (MRI) characterized these lesions as specific to fibrous dysplasia of the bone, without any insufficiency fracture at this level. The association of café-au-lait skin spots with bone fibrous dysplasia, and hyperthyroidism in this patient suggested the diagnosis of McCune - Albright syndrome.

  5. A Novel Detection Platform for Shrimp White Spot Syndrome Virus Using an ICP11-Dependent Immunomagnetic Reduction (IMR) Assay.

    PubMed

    Liu, Bing-Hsien; Lin, Yu-Chen; Ho, Chia-Shin; Yang, Che-Chuan; Chang, Yun-Tsui; Chang, Jui-Feng; Li, Chun-Yuan; Cheng, Cheng-Shun; Huang, Jiun-Yan; Lee, Yen-Fu; Hsu, Ming-Hung; Lin, Feng-Chun; Wang, Hao-Ching; Lo, Chu-Fang; Yang, Shieh-Yueh; Wang, Han-Ching

    2015-01-01

    Shrimp white spot disease (WSD), which is caused by white spot syndrome virus (WSSV), is one of the world's most serious shrimp diseases. Our objective in this study was to use an immunomagnetic reduction (IMR) assay to develop a highly sensitive, automatic WSSV detection platform targeted against ICP11 (the most highly expressed WSSV protein). After characterizing the magnetic reagents (Fe3O4 magnetic nanoparticles coated with anti ICP11), the detection limit for ICP11 protein using IMR was approximately 2 x 10(-3) ng/ml, and the linear dynamic range of the assay was 0.1~1 x 10(6) ng/ml. In assays of ICP11 protein in pleopod protein lysates from healthy and WSSV-infected shrimp, IMR signals were successfully detected from shrimp with low WSSV genome copy numbers. We concluded that this IMR assay targeting ICP11 has potential for detecting the WSSV.

  6. Minichromosome maintenance protein 7 regulates phagocytosis in kuruma shrimp Marsupenaeus japonicas against white spot syndrome virus.

    PubMed

    Wang, Zhi; Zhu, Fei

    2016-08-01

    Minichromosome maintenance protein (MCM7) belongs to the MCM protein family and participates in the MCM complex by playing a role in the cell replication cycle and chromosome initiation of eukaryotes. Previously, we found that several genes, including MCM7, were over-expressed in Drosophila melanogaster after white spot syndrome virus (WSSV) infection. In this study, we aimed to further research the MCM7 of kuruma shrimp, Marsupenaeus japonicus (mjMCM7) and determine its role in the innate immune system. To this end, we cloned the entire 2307-bp mjMCM7 sequence, including a 1974-bp open reading frame (ORF) encoding a 658-aa-long protein. Real-time PCR showed that the gene was primarily expressed in the hemolymph and hepatopancreas and over-expressed in shrimp challenged with WSSV. Gene function study was carried out by knocking down the expression of MCM7 using small interference RNA (siRNA). The results revealed that β-actin, hemocyanin, prophenoloxidase (proPO) and tumor necrosis factor-α (TNF-α) were up-regulated while the cytoskeleton proteins such as myosin and Rho were significantly down-regulated at 24 h after treatment. The results indicate a possible relationship between mjMCM7 and the innate immune system, and suggest that mjMCM7 may play a role in phagocytosis. After WSSV challenge, WSSV copies and mortality count were both higher in the MCM7-siRNA-treated groups at 60 h after treatment, and the mortality count approached that of the control groups over time. The phagocytosis rate was significantly lower in the MCM7-siRNA-treated group than in the WSSV group. The findings of this study confirm that mjMCM7 positively regulates phagocytosis and plays an important role against WSSV. These results could help researchers to further understand the function of the MCM7 protein and reveal its potential role in the innate immunity of invertebrates.

  7. Analysis of the complete nucleotide sequence of a white spot syndrome virus isolated from Pacific white shrimp.

    PubMed

    Chai, Choong Yee; Yoon, Jangmi; Lee, Yong Seok; Kim, Young Bong; Choi, Tae-Jin

    2013-10-01

    The fourth complete genome sequence of the white spot syndrome virus (WSSV) from Pacific white shrimp (Litopenaeus vannamei) isolated from Korea (WSSV-KR) was determined. The genome is composed of 295,884 bp encompassing 515 open reading frames (ORFs) among which 90 showed no sequence homology with any known protein in BLAST searches. The remaining 425 ORFs encode functional proteins including enzymes for nucleic acid metabolism, DNA replication and transcription, and several major structural proteins. Dot plot and linear comparisons of WSSV Korean strain with other WSSV isolates showed overall similarity but with some areas of sequence difference and one large deletion area.

  8. Arabidopsis-derived shrimp viral-binding protein, PmRab7 can protect white spot syndrome virus infection in shrimp.

    PubMed

    Thagun, Chonprakun; Srisala, Jiraporn; Sritunyalucksana, Kallaya; Narangajavana, Jarunya; Sojikul, Punchapat

    2012-09-15

    White spot syndrome virus is currently the leading cause of production losses in the shrimp industry. Penaeus monodon Rab7 protein has been recognized as a viral-binding protein with an efficient protective effect against white spot syndrome infection. Plant-derived recombinant PmRab7 might serve as an alternative source for in-feed vaccination, considering the remarkable abilities of plant expression systems. PmRab7 was introduced into the Arabidopsis thaliana T87 genome. Arabidopsis-derived recombinant PmRab7 showed high binding activity against white spot syndrome virus and a viral envelope, VP28. The growth profile of Arabidopsis suspension culture expressing PmRab7 (ECR21# 35) resembled that of its counterpart. PmRab7 expression in ECR21# 35 reached its maximum level at 5 mg g(-1) dry weight in 12 days, which was higher than those previously reported in Escherichia coli and in Pichia. Co-injection of white spot syndrome virus and Arabidopsis crude extract containing PmRab7 in Litopenaeus vannamei showed an 87% increase in shrimp survival rate at 5 day after injection. In this study, we propose an alternative PmRab7 source with higher production yield, and cheaper culture media costs, that might serve the industry's need for an in-feed supplement against white spot syndrome infection.

  9. Bacillus subtilis spores expressing the VP28 antigen: a potential oral treatment to protect Litopenaeus vannamei against white spot syndrome.

    PubMed

    Nguyen, Anh T V; Pham, Cuong K; Pham, Huong T T; Pham, Hang L; Nguyen, Anh H; Dang, Lua T; Huynh, Hong A; Cutting, Simon M; Phan, Tuan-Nghia

    2014-09-01

    The envelope protein VP28 of white spot syndrome virus (WSSV) is considered a candidate antigen for use in a potential vaccine to this important shrimp pathogen (the cause of white spot syndrome, WSS). Here, we used spores of Bacillus subtilis to display VP28 on the spore surface. Trials were conducted to evaluate their ability to protect shrimps against WSSV infection. The gene cotB-vp28 was integrated into the chromosome of the laboratory strain B. subtilis PY79, and expression of CotB-VP28 was detected by Western blotting and immunofluorescence. Expression of CotB-VP28 was equivalent to 1000 molecules per spore. PY79 and CotB-VP28 spores were mixed with pellets for feeding of whiteleg shrimps (Litopenaeus vannamei), followed by WSSV challenge. Superoxidase dismutase (SOD), phenoloxidase activities and mortality rates of the two shrimp groups were evaluated. Groups fed with PY79 and CotB-VP28 spores at day 7 had increased SOD activities of 29% and increased phenoloxidase activities of 15% and 33%, respectively, compared to those of the control group. Fourteen days postchallenge, 35% of vaccinated shrimps had died compared to 49% of those fed naked spores (PY79) and 66% untreated, unchallenged animals. These data suggest that spores expressing VP28 have potential as a prophylactic treatment of WSS.

  10. Comparison of protein expression profiles of the hepatopancreas in Fenneropenaeus chinensis challenged with heat-inactivated Vibrio anguillarum and white spot syndrome virus.

    PubMed

    Jiang, Hao; Li, Fuhua; Zhang, Jiquan; Zhang, Jinkang; Huang, Bingxin; Yu, Yang; Xiang, Jianhai

    2014-02-01

    Fenneropenaeus chinensis (Chinese shrimp) culture industry, like other Penaeidae culture, has been seriously affected by the shrimp diseases caused by bacteria and virus. To better understand the mechanism of immune response of shrimp to different pathogens, proteome research approach was utilized in this study. Firstly, the soluble hepatopancreas protein samples in adult Chinese shrimp among control, heat-inactivated Vibrio-challenged and white spot syndrome virus-infected groups were separated by 2-DE (pH range, 4-7; sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and pH range, 3-10; tricine-SDS-PAGE). Then the differentially expressed protein spots (≥1.5-fold or ≤0.67-fold averagely of controls) were analyzed by LC-ESI-MS/MS. Using Mascot online database searching algorithm and SEQUEST searching program, 48 and 49 differentially expressed protein spots were successfully identified in response to Vibrio and white spot syndrome virus infection, respectively. Based on these results, we discussed the mechanism of immune response of the shrimp and shed light on the differences between immune response of shrimp toward Vibrio and white spot syndrome virus. This study also set a basis for further analyses of some key genes in immune response of Chinese shrimp.

  11. Identification of the interaction domains of white spot syndrome virus envelope proteins VP28 and VP24.

    PubMed

    Li, Zaipeng; Chen, Weiyu; Xu, Limei; Li, Fang; Yang, Feng

    2015-03-16

    VP28 and VP24 are two major envelope proteins of white spot syndrome virus (WSSV). The direct interaction between VP28 and VP24 has been described in previous studies. In this study, we confirmed this interaction and mapped the interaction domains of VP28 and VP24 by constructing a series of deletion mutants. By co-immunoprecipitation, two VP28-binding domains of VP24 were located at amino acid residues 46-61 and 148-160, while VP24-binding domain of VP28 was located at amino acid residues 31-45. These binding domains were further corroborated by peptide blocking assay, in which synthetic peptides spanning the binding domains were able to inhibit VP28-VP24 interaction, whereas same-size control peptides from non-binging regions did not.

  12. Identification and characterization of a prawn white spot syndrome virus gene that encodes an envelope protein VP31

    SciTech Connect

    Li Li; Xie Xixian; Yang Feng . E-mail: mbiotech@public.xm.fj.cn

    2005-09-15

    Based on a combination of SDS-PAGE and mass spectrometry, a protein with an apparent molecular mass of 31 kDa (termed as VP31) was identified from purified shrimp white spot syndrome virus (WSSV) envelope fraction. The resulting amino acid (aa) sequence matched an open reading frame (WSV340) of the WSSV genome. This ORF contained 783 nucleotides (nt), encoding 261 aa. A fragment of WSV340 was expressed in Escherichia coli as a glutathione S-transferase (GST) fusion protein with a 6His-tag, and then specific antibody was raised. Western blot analysis and the immunoelectron microscope method (IEM) confirmed that VP31 was present exclusively in the viral envelope fraction. The neutralization experiment suggested that VP31 might play an important role in WSSV infectivity.

  13. Increasing production in Korean shrimp farms with white-spot syndrome virus PCR-negative brood stock.

    PubMed

    Seok, Hyeok Seung; Baek, Min Won; Lee, Hui Young; Kim, Dong Jae; Chun, Myung Sun; Kim, Jong Sheek; Chang, Se Ok; Park, Jae Hak

    2007-03-01

    White-spot syndrome virus (WSSV) is a devastating, infectious virus affecting shrimp. Although sensitive techniques involving PCR have been developed to assist farmers in screening shrimp (brood stock) for WSSV prior to stocking ponds, such practices have not yet been applied in Korea. Despite the rationality of implementing screening, there has been some doubt as to whether the stocking of WSSV-PCR-negative fry epidemiologically decreases white-spot disease outbreaks. Here, we report a retrospective analysis of data from shrimp farms in the western coast of Korea where WSSV-PCR-negative brood stocks were used to stock rearing ponds. A total of 366 shrimp from Heuksan Island were sampled for WSSV with PCR. Of the tested shrimp, 7.2% (28 brood stocks) were identified as WSSV positive; only WSSV-PCR-negative shrimp were used for brood stocks. Total unit production (final shrimp production/ the area of the ponds) was higher, at 1.96, in ponds where WSSV-PCR-negative shrimp were used, as compared with 1.02 in other ponds in Korea in 2004. This retrospective analysis of WSSV in Korea may be useful to the shrimp aquaculture industry, suggesting a testable hypothesis that may contribute to the eventual control of WSSV outbreaks.

  14. Two White Spot Syndrome Virus MicroRNAs Target the Dorsal Gene To Promote Virus Infection in Marsupenaeus japonicus Shrimp.

    PubMed

    Ren, Qian; Huang, Xin; Cui, Yalei; Sun, Jiejie; Wang, Wen; Zhang, Xiaobo

    2017-04-15

    In eukaryotes, microRNAs (miRNAs) serve as regulators of many biological processes, including virus infection. An miRNA can generally target diverse genes during virus-host interactions. However, the regulation of gene expression by multiple miRNAs has not yet been extensively explored during virus infection. This study found that the Spaztle (Spz)-Toll-Dorsal-antilipopolysaccharide factor (ALF) signaling pathway plays a very important role in antiviral immunity against invasion of white spot syndrome virus (WSSV) in shrimp (Marsupenaeus japonicus). Dorsal, the central gene in the Toll pathway, was targeted by two viral miRNAs (WSSV-miR-N13 and WSSV-miR-N23) during WSSV infection. The regulation of Dorsal expression by viral miRNAs suppressed the Spz-Toll-Dorsal-ALF signaling pathway in shrimp in vivo, leading to virus infection. Our study contributes novel insights into the viral miRNA-mediated Toll signaling pathway during the virus-host interaction.IMPORTANCE An miRNA can target diverse genes during virus-host interactions. However, the regulation of gene expression by multiple miRNAs during virus infection has not yet been extensively explored. The results of this study indicated that the shrimp Dorsal gene, the central gene in the Toll pathway, was targeted by two viral miRNAs during infection with white spot syndrome virus. Regulation of Dorsal expression by viral miRNAs suppressed the Spz-Toll-Dorsal-ALF signaling pathway in shrimp in vivo, leading to virus infection. Our study provides new insight into the viral miRNA-mediated Toll signaling pathway in virus-host interactions.

  15. Gene expression profiling in gill tissues of White spot syndrome virus infected black tiger shrimp Penaeus monodon by DNA microarray.

    PubMed

    Shekhar, M S; Gomathi, A; Gopikrishna, G; Ponniah, A G

    2015-06-01

    White spot syndrome virus (WSSV) continues to be the most devastating viral pathogen infecting penaeid shrimp the world over. The genome of WSSV has been deciphered and characterized from three geographical isolates and significant progress has been made in developing various molecular diagnostic methods to detect the virus. However, the information on host immune gene response to WSSV pathogenesis is limited. Microarray analysis was carried out as an approach to analyse the gene expression in black tiger shrimp Penaeus monodon in response to WSSV infection. Gill tissues collected from the WSSV infected shrimp at 6, 24, 48 h and moribund stage were analysed for differential gene expression. Shrimp cDNAs of 40,059 unique sequences were considered for designing the microarray chip. The Cy3-labeled cRNA derived from healthy and WSSV-infected shrimp was subjected to hybridization with all the DNA spots in the microarray which revealed 8,633 and 11,147 as up- and down-regulated genes respectively at different time intervals post infection. The altered expression of these numerous genes represented diverse functions such as immune response, osmoregulation, apoptosis, nucleic acid binding, energy and metabolism, signal transduction, stress response and molting. The changes in gene expression profiles observed by microarray analysis provides molecular insights and framework of genes which are up- and down-regulated at different time intervals during WSSV infection in shrimp. The microarray data was validated by Real Time analysis of four differentially expressed genes involved in apoptosis (translationally controlled tumor protein, inhibitor of apoptosis protein, ubiquitin conjugated enzyme E2 and caspase) for gene expression levels. The role of apoptosis related genes in WSSV infected shrimp is discussed herein.

  16. Localization of VP28 on the baculovirus envelope and its immunogenicity against white spot syndrome virus in Penaeus monodon

    SciTech Connect

    Syed Musthaq, S.; Madhan, Selvaraj; Sahul Hameed, A.S.; Kwang, Jimmy

    2009-09-01

    White spot syndrome virus (WSSV) is a large dsDNA virus responsible for white spot disease in shrimp and other crustaceans. VP28 is one of the major envelope proteins of WSSV and plays a crucial role in viral infection. In an effort to develop a vaccine against WSSV, we have constructed a recombinant baculovirus with an immediate early promoter 1 which expresses VP28 at an early stage of infection in insect cells. Baculovirus expressed rVP28 was able to maintain its structural and antigenic conformity as indicated by immunofluorescence assay and western blot analysis. Interestingly, our results with confocal microscopy revealed that rVP28 was able to localize on the plasma membrane of insect cells infected with recombinant baculovirus. In addition, we demonstrated with transmission electron microscopy that baculovirus successfully acquired rVP28 from the insect cell membrane via the budding process. Using this baculovirus displaying VP28 as a vaccine against WSSV, we observed a significantly higher survival rate of 86.3% and 73.5% of WSSV-infected shrimp at 3 and 15 days post vaccination respectively. Quantitative real-time PCR also indicated that the WSSV viral load in vaccinated shrimp was significantly reduced at 7 days post challenge. Furthermore, our RT-PCR and immunohistochemistry results demonstrated that the recombinant baculovirus was able to express VP28 in vivo in shrimp tissues. This study will be of considerable significance in elucidating the morphogenesis of WSSV and will pave the way for new generation vaccines against WSSV.

  17. Differential gene expression profile of the hepatopancreas of white spot syndrome virus infected Fenneropenaeus indicus by suppression subtractive hybridization.

    PubMed

    James, Roswin; Thampuran, Nirmala; Lalitha, K V; Rajan, Lawrance Anbu; Joseph, Toms C

    2010-11-01

    Suppression Subtractive Hybridization was employed in order to identify the differentially expressed genes in the hepatopancreas of white spot syndrome virus infected Fenneropenaeus indicus. A forward subtracted cDNA library generated 356 clones following a white spot syndrome virus infection. A total of 345 clones with more than 100 nucleotides were selected for further analysis using bioinformatics tools after vector screening. Twenty-three contigs and 111 singletons were generated from a total of 134 consensuses. The consensuses, on a sequence homology search using BLASTX (NCBI), revealed that 74 (55%) of them had no significant match to reported sequences in the database, suggesting that they were found for the first time and are probably associated with shrimp immune function. Out of the remaining 60 (45%) consensuses, 43 had significant homology to known protein sequences in the database while 17 consensuses are homologous to unknown proteins in the database which are considered novel. The most abundant genes in the subtracted library were antimicrobial peptides accounting for 56 clones; among which one is a member of SNF2 family of proteins and another belonged to PfP1 family of proteins on analysis using Antimicrobial peptide predictor software. The other predicted genes in the subtracted library include signal transduction molecules (GTPase, Serine threonine kinase, Armadillo repeats etc), antioxidant enzymes (Cytochrome oxidase, Monomeric sarcosine oxidase and Catalase), active transporters (Nuclear Localization Signal [NLS], calcium ATPase, sodium glutamate symporter, Store-Operated Calcium Entry [SOCE] and ribonucleoprotein [RNP]) contributing to 19, 14 and 5 clones respectively. Three clones are homologous to reverse transcriptase; a first time report in shrimp and one each belong to cell adhesion molecule and Proteinase. InterProScan at EMBL, when used for an integrated search at PROSITE predicted; signal sequences and transmembrane regions for 13

  18. Relationship between white spot syndrome virus (WSSV) loads and characterizations of water quality in Litopenaeus vannamei culture ponds during the tropical storm.

    PubMed

    Zhang, J S; Li, Z J; Wen, G L; Wang, Y L; Luo, L; Zhang, H J; Dong, H B

    2016-01-01

    An in-situ experiment was conducted to investigate the effect of tropical storm on the white spot syndrome virus (WSSV) loads in Litopenaeus vannamei rearing ponds. White spot syndrome virus loads, heterotrophic bacteria, Vibrio and water quality (including temperature, dissolved oxygen (DO), salinity, pH, NH4-N, and NO2-N) were continually monitored through one tropical storm. The WSSV loads decreased when tropical storm made landfall, and substantially increased when typhoon passed. The variation of WSSV loads was correlated with DO, temperature, heterotrophic bacteria count, and ammonia-N concentrations. These results suggested that maintaining high level DO and promoting heterotrophic bacteria growth in the shrimp ponds might prevent the diseases' outbreak after the landfall of tropical storm.

  19. Relationship between white spot syndrome virus (WSSV) loads and characterizations of water quality in Litopenaeus vannamei culture ponds during the tropical storm

    PubMed Central

    Zhang, J. S.; Li, Z. J.; Wen, G. L.; Wang, Y. L.; Luo, L.; Zhang, H. J.; Dong, H. B.

    2016-01-01

    An in-situ experiment was conducted to investigate the effect of tropical storm on the white spot syndrome virus (WSSV) loads in Litopenaeus vannamei rearing ponds. White spot syndrome virus loads, heterotrophic bacteria, Vibrio and water quality (including temperature, dissolved oxygen (DO), salinity, pH, NH4-N, and NO2-N) were continually monitored through one tropical storm. The WSSV loads decreased when tropical storm made landfall, and substantially increased when typhoon passed. The variation of WSSV loads was correlated with DO, temperature, heterotrophic bacteria count, and ammonia-N concentrations. These results suggested that maintaining high level DO and promoting heterotrophic bacteria growth in the shrimp ponds might prevent the diseases’ outbreak after the landfall of tropical storm. PMID:27822254

  20. White spot syndrome virus isolates of tiger shrimp Penaeus monodon (Fabricious) in India are similar to exotic isolates as revealed by polymerase chain reaction and electron microscopy.

    PubMed

    Mishra, S S; Shekhar, M S

    2005-07-01

    Microbiological analysis of samples collected from cases of white spot disease outbreaks in cultured shrimp in different farms located in three regions along East Coast of India viz. Chidambram (Tamil Nadu), Nellore (Andhra Pradesh) and Balasore (Orissa), revealed presence of Vibrio alginolyticus, Vibrio parahaemolyticus, and Aeromonas spp. but experimental infection trials in Penaeus monodon with these isolates did not induce any acute mortality or formation of white spots on carapace. Infection trials using filtered tissue extracts by oral and injection method induced mortality in healthy P. monodon with all samples and 100% mortality was noted by the end of 7 day post-inoculation. Histopathological analysis demonstrated degenerated cells characterized by hypertrophied nuclei in gills, hepatopancreas and lymphoid organ with presence of intranuclear basophilic or eosino-basophilic bodies in tubular cells and intercellular spaces. Analysis of samples using 3 different primer sets as used by other for detection of white spot syndrome virus (WSSV) generated 643, 1447 and 520bp amplified DNA products in all samples except in one instance. Variable size virions with mean size in the range of 110 x 320 +/- 20 nm were observed under electron microscope. It could be concluded that the viral isolates in India involved with white spot syndrome in cultured shrimp are similar to RV-PJ and SEMBV in Japan, WSBV in Taiwan and WSSV in Malaysia, Indonesia, Thailand, China and Japan.

  1. Microarray and RT-PCR screening for white spot syndrome virus immediate-early genes in cycloheximide-treated shrimp

    SciTech Connect

    Liu Wangjing; Chang Yunshiang; Wang Chunghsiung; Kou, Guang-Hsiung; Lo Chufang . E-mail: gracelow@ntu.edu.tw

    2005-04-10

    Here, we report for the first time the successful use of cycloheximide (CHX) as an inhibitor to block de novo viral protein synthesis during WSSV (white spot syndrome virus) infection. Sixty candidate IE (immediate-early) genes were identified using a global analysis microarray technique. RT-PCR showed that the genes corresponding to ORF126, ORF242 and ORF418 in the Taiwan isolate were consistently CHX-insensitive, and these genes were designated ie1, ie2 and ie3, respectively. The sequences for these IE genes also appear in the two other WSSV isolates that have been sequenced. Three corresponding ORFs were identified in the China WSSV isolate, but only an ORF corresponding to ie1 was predicted in the Thailand isolate. In a promoter activity assay in Sf9 insect cells using EGFP (enhanced green fluorescence protein) as a reporter, ie1 showed very strong promoter activity, producing higher EGFP signals than the insect Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (OpMNPV) ie2 promoter.

  2. Identification of differentially expressed genes in shrimp (Penaeus stylirostris) infected with White spot syndrome virus by cDNA microarrays.

    PubMed

    Dhar, A K; Dettori, A; Roux, M M; Klimpel, K R; Read, B

    2003-12-01

    White spot syndrome virus (WSSV) is currently the most important viral pathogen infecting penaeid shrimp worldwide. Although considerable progress has been made in characterizing the WSSV genome and developing detection methods, information pertaining to host genes involved in WSSV pathogenesis is limited. We examined the potential of cDNA microarray analysis to study gene expression in WSSV-infected shrimp. Shrimp cDNAs were printed as low-density arrays on glass slides and were hybridized with Cy3/Cy5 labeled probes derived from RNA isolated from healthy and WSSV-infected shrimp. Genes that code for proteins that are relevant to crustacean immunity, structural proteins, as well as proteins of unknown function were among those whose mRNA expression was altered upon WSSV infection. To validate the microarray data, the temporal expression of three differentially expressed genes, an immune gene (C-type lectin-1), a structural gene (40S ribosomal protein), and a gene involved in lipid metabolism (fatty acid binding protein) was measured in healthy and WSSV-infected shrimp by real-time RT-PCR. The data suggest that WSSV infection alters the expression of a wide array of cellular genes, and provides a framework for further studies aimed at identifying genes whose function may provide insight into the mechanism of WSSV infection in shrimp.

  3. Mixed-genotype white spot syndrome virus infections of shrimp are inversely correlated with disease outbreaks in ponds.

    PubMed

    Hoa, Tran Thi Tuyet; Zwart, Mark P; Phuong, Nguyen T; Oanh, Dang T H; de Jong, Mart C M; Vlak, Just M

    2011-03-01

    Outbreaks of white spot syndrome virus (WSSV) in shrimp culture and the relationship between the virus and virulence are not well understood. Here, we provide evidence showing that WSSV mixed-genotype infections correlate with lower outbreak incidence and that disease outbreaks correlate with single-genotype infections. We tested 573 shrimp samples from 81 shrimp ponds in the Mekong delta with outbreak or non-outbreak status. The variable number tandem repeat (VNTR) loci of WSSV were used as molecular markers for the characterization of single- and mixed-genotype infections. The overall prevalence of mixed-genotype WSSV infections was 25.7 %. Non-outbreak ponds had a significantly higher frequency of mixed-genotype infections than outbreak ponds for all VNTR loci, both at the individual shrimp as well as at the pond level. The genetic composition of WSSV populations appears to correlate with the health status of shrimp culture in ponds. The causal relationship between genotypic diversity and disease outbreaks can now be experimentally approached.

  4. Collaboration between a soluble C-type lectin and calreticulin facilitates white spot syndrome virus infection in shrimp.

    PubMed

    Wang, Xian-Wei; Xu, Yi-Hui; Xu, Ji-Dong; Zhao, Xiao-Fan; Wang, Jin-Xing

    2014-09-01

    White spot syndrome virus (WSSV) mainly infects crustaceans through the digestive tract. Whether C-type lectins (CLs), which are important receptors for many viruses, participate in WSSV infection in the shrimp stomach remains unknown. In this study, we orally infected kuruma shrimp Marsupenaeus japonicus to model the natural transmission of WSSV and identified a CL (designated as M. japonicus stomach virus-associated CL [MjsvCL]) that was significantly induced by virus infection in the stomach. Knockdown of MjsvCL expression by RNA interference suppressed the virus replication, whereas exogenous MjsvCL enhanced it. Further analysis by GST pull-down and coimmunoprecipitation showed that MjsvCL could bind to viral protein 28, the most abundant and functionally relevant envelope protein of WSSV. Furthermore, cell-surface calreticulin was identified as a receptor of MjsvCL, and the interaction between these proteins was a determinant for the viral infection-promoting activity of MjsvCL. The MjsvCL-calreticulin pathway facilitated virus entry likely in a cholesterol-dependent manner. This study provides insights into a mechanism by which soluble CLs capture and present virions to the cell-surface receptor to facilitate viral infection.

  5. Emilia sonchifolia extract activity against white spot syndrome virus and yellow head virus in shrimp cell cultures.

    PubMed

    Maikaeo, Lamai; Chotigeat, Wilaiwan; Mahabusarakam, Wilawan

    2015-07-23

    Emilia sonchifolia (L.) DC is a plant used in traditional medicine to treat several viral and bacterial diseases. The antiviral activities of selected Sephadex LH-20 column fractions and HPLC subfractions of an acetone extract of E. sonchifolia leaves were determined in shrimp Penaeus merguiensis primary lymphoid cells infected with either white spot syndrome virus (WSSV) or yellow head virus (YHV). WSSV and YHV replication was quantified using quantitative real-time PCR tests targeted to the VP19 and ORF1b gene transcripts, respectively. In lymphoid organ cells exposed to 100 µg ml⁻¹ of either the Sephadex fraction F14 or the HPLC F14 subfraction SF4, both fractions caused reduced replication, but YHV replication was reduced only by SF4. In the asthiazolyl blue mitochondrial enzyme activity assays to assess extract cytotoxicity, >60% of primary lymphoid organ cells remained viable following exposure to 100 µg ml⁻¹ of either F14 or SF4. GC-MS analysis of the HPLC F14 subfraction SF4 showed that it contained 2,4-di-tert-butylphenol. This study is the first to show that E. sonchifolia leaf extracts might be useful as bioactive agents to protect shrimp against viruses such as WSSV and YHV.

  6. Molecular modeling and expression of the Litopenaeus vannamei proliferating cell nuclear antigen (PCNA) after white spot syndrome virus shrimp infection.

    PubMed

    de-la-Re-Vega, Enrique; Muhlia-Almazan, Adriana; Arvizu-Flores, Aldo A; Islas-Osuna, Maria A; Yepiz-Plascencia, Gloria; Brieba, Luis G; Sotelo-Mundo, Rogerio R

    2011-01-01

    Proliferating cell nuclear antigen (PCNA) is the eukaryotic sliding clamp that tethers DNA polymerase to DNA during replication. The full-length cDNA of the Pacific white shrimp Litopenaeus vannamei PCNA (LvPCNA) was cloned and encoded a protein of 260 amino acids that is highly similar to other Crustacean PCNAs. The theoretical shrimp PCNA structure has all the domains that are necessary for its interaction with template DNA and DNA polymerase. RT-PCR analysis showed that LvPCNA is expressed mainly in muscle and hemocytes and much less in hepatopancreas and gills. LvPCNA mRNA levels are not statistically different in muscle from healthy and challenged shrimp with the white spot syndrome virus (WSSV). In contrast, the mRNA levels of the viral DNA polymerase show a biphasic pattern with expression at 6 h post-infection and later at 24 and 48 h. These results suggest that in shrimp muscle LvPCNA levels are steadily kept to allow viral replication and that WSSV DNA polymerase (WSSV-DNApol) is more responsive towards later stages of infection. More knowledge of the DNA replication machinery would result in a better understanding of the mechanism and components of viral replication, since the WSSV genome does not have all the components required for assembly of a fully functional replisome.

  7. An immediate-early protein of white spot syndrome virus modulates the phosphorylation of focal adhesion kinase of shrimp.

    PubMed

    Lu, Huasong; Ruan, Lingwei; Xu, Xun

    2011-10-25

    WSSV interacts with integrin during infection of shrimps and modulate the focal adhesion kinase which is known as a regulator of several downstream signaling pathways. Viral protein kinases are thought to be important for virus infection by regulating the host signaling pathways. WSV083 is an immediate-early gene of white spot syndrome virus that contains a Ser/Thr protein kinase domain. So, does WSSV modulate FAK phosphorylation via the WSV083 molecule? In this study, co-transfection of WSV083 and MjFAK genes proceeded in insect cells revealed that the MjFAK phosphorylation and cell adhesion activity could be inhibited by the expression of WSV083. Kinase domain mutants of WSV083 lost its ability of inhibiting FAK phosphorylation. Moreover, silencing of FAK gene through RNAi accelerated the shrimp death rate upon WSSV challenge. These results demonstrate for the first time that modulation of FAK phosphorylation by WSV083 plays a critical role in the pathogenesis of WSSV infection.

  8. Label free detection of white spot syndrome virus using lead magnesium niobate-lead titanate piezoelectric microcantilever sensors.

    PubMed

    Capobianco, Joseph A; Shih, Wei-Heng; Leu, Jiann-Horng; Lo, Grace Chu-Fang; Shih, Wan Y

    2010-11-15

    We have investigated rapid, label free detection of white spot syndrome virus (WSSV) using the first longitudinal extension resonance peak of five lead-magnesium niobate-lead titanate (PMN-PT) piezoelectric microcantilever sensors (PEMS) 1050-700 μm long and 850-485 μm wide constructed from 8 μm thick PMN-PT freestanding films. The PMN-PT PEMS were encapsulated with a 3-mercaptopropyltrimethoxysilane (MPS) insulation layer and further coated with anti-VP28 and anti-VP664 antibodies to target the WSSV virions and nucleocapsids, respectively. By inserting the antibody coated PEMS in a flowing virion or nucleocapsid suspension, label free detection of the virions and nucleocapsids were respectively achieved by monitoring the PEMS resonance frequency shift. We showed that positive label free detection of both the virion and the nucleocapsid could be achieved at a concentration of 100virions(nucleocapsids)/ml or 10 virions(nucleocapsids)/100 μl, comparable to the detection sensitivity of polymerase chain reaction (PCR). However, in contrast to PCR, PEMS detection was label free, in situ and rapid (less than 30 min), potentially requiring minimal or no sample preparation.

  9. Dorsal transcription factor is involved in regulating expression of crustin genes during white spot syndrome virus infection.

    PubMed

    Huang, Xin; Wang, Wen; Ren, Qian

    2016-10-01

    Nuclear factor-kappa B (NF-κB) pathways play important roles in innate immune responses. In this study, we identified a dorsal homolog (MrDorsal) from freshwater prawn Macrobrachium rosenbergii. The full-length cDNA of MrDorsal comprised 2533 bp with an open reading frame of 1986 bp, which encoded a peptide of 661 amino acid residues. Amino acid sequence analysis showed that MrDorsal contains a Rel homolog domain and an IPT/TIG (i.e., Ig-like, plexin, and transcription factors) domain. The signature sequence of dorsal protein FRYMCEG existed in the deduced amino acid sequence. Sequence analysis showed that MrDorsal shared high similarities with Dorsal from invertebrate species. MrDorsal was abundant in the hemocytes and gills of healthy prawns but minute levels were detected in other tissues. The expression of MrDorsal was significantly upregulated 48 h after the white spot syndrome virus (WSSV-) challenge. Knockdown of MrDorsal using double-stranded RNA could suppress the transcription of crustin genes (MrCrustin2 and MrCrustin4) in gills of prawns after 48 h of the WSSV challenge. Results indicated that MrDorsal was involved to regulate the expression of crustin genes and it might play potential important roles during WSSV infection.

  10. Antigenic and immunogenic properties of truncated VP28 protein of white spot syndrome virus in Procambarus clarkii.

    PubMed

    Du, Hua-Hua; Hou, Chong-Lin; Wu, Xiao-Guo; Xie, Rong-hui; Wang, Yi-Zhen

    2013-01-01

    Previous studies identify VP28 envelope protein of white spot syndrome virus (WSSV) as its main antigenic protein. Although implicated in viral infectivity, its functional role remains unclear. In the current study, we described the production of polyclonal antibodies to recombinant truncated VP28 proteins including deleted N-terminal (rVP28ΔN), C-terminal (rVP28ΔC) and middle (rVP28ΔM). In antigenicity assays, antibodies developed from VP28 truncations lacking the N-terminal or middle regions showed significantly lowered neutralization of WSSV in crayfish, Procambarus clarkii. Further immunogenicity analysis showed reduced relative percent survival (RPS) in crayfish vaccinating with these truncations before challenge with WSSV. These results indicated that N-terminal (residues 1-27) and middle region (residues 35-95) were essential to maintain the neutralizing linear epitopes of VP28 and responsible in eliciting immune response. Thus, it is most likely that these regions are exposed on VP28, and will be useful for rational design of effective vaccines targeting VP28 of WSSV.

  11. Application of a Label-Free Immunosensor for White Spot Syndrome Virus (WSSV) in Shrimp Cultivation Water.

    PubMed

    Waiyapoka, Thanyaporn; Deachamag, Panchalika; Chotigeat, Wilaiwan; Bunsanong, Nittaya; Kanatharana, Proespichaya; Thavarungkul, Panote; Loyprasert-Thananimit, Suchera

    2015-10-01

    White spot syndrome virus (WSSV) is a major pathogen affecting the shrimp industry worldwide. In a preliminary study, WSSV binding protein (WBP) was specifically bound to the VP26 protein of WSSV. Therefore, we have developed the label-free affinity immunosensor using the WBP together with anti-GST-VP26 for quantitative detection of WSSV in shrimp pond water. When the biological molecules were immobilized on a gold electrode to form a self-assembled monolayer, it was then used to detect WSSV using a flow injection system with optimized conditions. Binding between the different copies of WSSV and the immobilized biological molecules was detected by an impedance change (ΔZ″) in real time. The sensitivity of the developed immunosensor was in the linear range of 1.6 × 10(1)-1.6 × 10(6) copies/μl. The system was highly sensitive for the analysis of WSSV as shown by the lack of impedance change when using yellow head virus (YHV). The developed immunosensor could be reused up to 37 times (relative standard deviation (RSD), 3.24 %) with a good reproducibility of residual activity (80-110 %). The immunosensor was simple to operate, reliable, reproducible, and could be applied for the detection and quantification of WSSV in water during shrimp cultivation.

  12. Molecular characterization, immune response against white spot syndrome virus infection of peroxiredoxin 4 in Fenneropenaeus chinensis and its antioxidant activity.

    PubMed

    Zhang, Qingli; Huang, Jie; Li, Fuhua; Liu, Shuang; Liu, Qinghui; Wei, Jiankai; Liang, Gaofeng; Xiang, Jianhai

    2014-03-01

    Peroxiredoxins (Prx) are a family of antioxidant proteins and perform important functions in intracellular signal transduction. Here, we report a Prx gene from Chinese shrimp Fenneropenaeus chinensis. The full-length cDNA of FcPrx gene contained an open reading frame of 735 bp encoding a polypeptide of 275 amino acids. The molecular mass of the deduced amino acid of FcPrx is 27445.43 Da with an estimated pI of 5.71. Sequence comparison showed that the FcPrx shares high identities with Prx IVs and it was named FcPrx4. A real-Time PCR (qRT-PCR) assay was developed to assess the mRNA expression of FcPrx4 in different tissues and temporal expression in hemocytes and hepatopancreas of F. chinensis challenged by white spot syndrome virus (WSSV). Transcripts of FcPrx4 can be detected in all tissues examined. The expression of FcPrx4 showed significant up-regulation in shrimp hemocytes and hepatopancreas after artificial infection with WSSV. A fusion protein containing FcPrx4 was produced in vitro and was confirmed by Matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) assay. And activity analysis indicated that the recombinant FcPrx4 proteins can reduce H2O2 in the presence of dithiothreitol.

  13. Enzyme E2 from Chinese white shrimp inhibits replication of white spot syndrome virus and ubiquitinates its RING domain proteins.

    PubMed

    Chen, An-Jing; Wang, Shuai; Zhao, Xiao-Fan; Yu, Xiao-Qiang; Wang, Jin-Xing

    2011-08-01

    Recent studies have shown that the ubiquitin (Ub) proteasome pathway (UPP) is closely related to immune defense. We have identified a ubiquitin-conjugating enzyme, E2, from the Chinese white shrimp, Fenneropenaeus chinensis (FcUbc). Injection of recombinant FcUbc protein (rFcUbc) reduced the mortality of shrimp infected with white spot syndrome virus (WSSV) and inhibited replication of WSSV. rFcUbc, but not a mutant FcUbc (mFcUbc), bound to WSSV RING domains (WRDs) from four potential E3 ligase proteins of WSSV in vitro. Importantly, rFcUbc could ubiquitinate the RING domains (named WRD2 and WRD3) of WSSV277 and WSSV304 proteins in vitro and the two proteins in WSSV-infected Drosophila melanogaster Schneider 2 (S2) cells. Furthermore, overexpression of FcUbc increased ubiquitination of WSSV277 and WSSV304 during WSSV infection. In summary, our study demonstrates that FcUbc from Chinese white shrimp inhibited WSSV replication and could ubiquitinate WSSV RING domain-containing proteins. This is the first report about antiviral function of Ubc E2 in shrimp.

  14. A study of the role of glucose transporter 1 (Glut1) in white spot syndrome virus (WSSV) infection.

    PubMed

    Huang, Huai-Ting; Chan, Hoi-Ling; Shih, Tsai-Yen; Chen, Li-Li

    2015-10-01

    White spot syndrome virus (WSSV) is a large enveloped DNA virus, and it causes a serious disease that has led to severe mortalities of cultured shrimps in many countries. To determine the mechanism of virus entry into the cell and to establish an antiviral strategy, the cell receptor for virus entry and receptor binding protein should be identified. A shrimp cell surface protein, glucose transporter1 (Glut1), was found to interact with WSSV in previous study. In this study, this Glut1 was confirmed to have the ability of transporting glucose, and this gene can also be found in other shrimp species. The interaction between Glut1 and some other WSSV envelope proteins in the infectome structure was verified by far western blot and His pull down assay. In vitro and in vivo neutralization using recombinant partial Glut1 revealed that the large extracellular portion of Glut1 could delay WSSV infection. Also, shrimps which were knocked-down Glut1 gene by treated with dsRNA before WSSV challenge showed decreased mortality. These results indeed provide a direction to develop efficient antiviral strategies or therapeutic methods by using Glut1.

  15. A sensitive and specific hyperbranched rolling circle amplification assay and test strip for white spot syndrome virus.

    PubMed

    Zhao, Yu-Ran; Yin, Wei-Li; Yue, Zhi-Qin; Li, Ba-Fang

    2014-01-01

    White spot syndrome virus (WSSV) is a global threat to the prawn industry, and there is no simple method for field-based testing of this virus. We designed a padlock probe and primers to the capsid protein gene VP28 of WSSV, and established a hyperbranched rolling circle amplification (HRCA) assay and a corresponding strip-based test. The assay and the test strip both had similar high accuracy and specificity, and their sensitivity was about 10 copies/μL, which is 100 times higher than conventional PCR. In this study, 68 batches of prawns were tested for WSSV with the HRCA assay and test strip, and the results were compared with the PCR assay. The results indicated that both the assay and test strip had accuracy similar to each other and to the PCR results. However, the assay and strip were more sensitive and user-friendly than PCR. Establishment of this method will provide a rapid detection of WSSV and also a basis for field-based detection of animal disease.

  16. High efficacy of white spot syndrome virus replication in tissues of freshwater rice-field crab, Paratelphusa hydrodomous (Herbst).

    PubMed

    Sundar Raj, N; Nathiga Nambi, K S; Abdul Majeed, S; Taju, G; Vimal, S; Farook, M A; Sahul Hameed, A S

    2012-12-01

    An attempt was made to determine the replication efficiency of white spot syndrome virus (WSSV) of shrimp in different organs of freshwater rice-field crab, Paratelphusa hydrodomous (Herbst), using bioassay, PCR, RT-PCR, ELISA, Western blot and real-time PCR analyses, and also to use this crab instead of penaeid shrimp for the large-scale production of WSSV. This crab was found to be highly susceptible to WSSV by intramuscular injection. PCR and Western blot analyses confirmed the systemic WSSV infection in freshwater crab. The RT-PCR analysis revealed the expression of VP28 gene in different organs of infected crab. The indirect ELISA was used to quantify the VP28 protein in different organs of crab. It was found that there was a high concentration of VP28 protein in gill tissue, muscle, haemolymph and heart tissue. The copy number of WSSV in different organs of infected crab was quantified by real-time PCR, and the results revealed a steady increase in copy number in different organs of infected crab during the course of infection. The viral inoculum prepared from different organs of infected crab caused significant mortality in tiger prawn, Penaeus monodon (Fabricius). The results revealed that this crab can be used as an alternate host for WSSV replication and production.

  17. Graphene oxide based fluorescence resonance energy transfer and loop-mediated isothermal amplification for white spot syndrome virus detection.

    PubMed

    Waiwijit, U; Phokaratkul, D; Kampeera, J; Lomas, T; Wisitsoraat, A; Kiatpathomchai, W; Tuantranont, A

    2015-10-20

    Graphene oxide (GO) is attractived for biological or medical applications due to its unique electrical, physical, optical and biological properties. In particular, GO can adsorb DNA via π-π stacking or non-covalent interactions, leading to fluorescence quenching phenomenon applicable for bio-molecular detection. In this work, a new method for white spot syndrome virus (WSSV)-DNA detection is developed based on loop-mediated isothermal amplification (LAMP) combined with fluorescence resonance energy transfer (FRET) between GO and fluorescein isothiocyanate-labeled probe (FITC-probe). The fluorescence quenching efficiency of FITC-probe was found to increase with increasing GO concentration and reached 98.7% at a GO concentration of 50 μg/ml. The fluorescence intensity of FITC-probe was recovered after hybridization with WSSV LAMP product with an optimal hybridization time of 10 min and increased accordingly with increasing amount of LAMP products. The detection limit was estimated to be as low as 10 copies of WSSV plasmid DNA or 0.6 fg of the total DNA extracted from shrimp infected with WSSV. In addition, no cross reaction was observed with other common shrimp viral pathogens. Therefore, the GO-FRET-LAMP technique is promising for fast, sensitive and specific detection of DNAs.

  18. White spot syndrome virus (WSSV) genome stability maintained over six passages through three different penaeid shrimp species.

    PubMed

    Sindhupriya, M; Saravanan, P; Otta, S K; Amarnath, C Bala; Arulraj, R; Bhuvaneswari, T; Praveena, P Ezhil; Jithendran, K P; Ponniah, A G

    2014-08-21

    White spot syndrome virus (WSSV) replicates rapidly, can be extremely pathogenic and is a common cause of mass mortality in cultured shrimp. Variable number tandem repeat (VNTR) sequences present in the open reading frame (ORF)94, ORF125 and ORF75 regions of the WSSV genome have been used widely as genetic markers in epidemiological studies. However, reports that VNTRs might evolve rapidly following even a single transmission through penaeid shrimp or other crustacean hosts have created confusion as to how VNTR data is interpreted. To examine VNTR stability again, 2 WSSV strains (PmTN4RU and LvAP11RU) with differing ORF94 tandem repeat numbers and slight differences in apparent virulence were passaged sequentially 6 times through black tiger shrimp Penaeus monodon, Indian white shrimp Feneropenaeus indicus or Pacific white leg shrimp Litopenaeus vannamei. PCR analyses to genotype the ORF94, ORF125 and ORF75 VNTRs did not identify any differences from either of the 2 parental WSSV strains after multiple passages through any of the shrimp species. These data were confirmed by sequence analysis and indicate that the stability of the genome regions containing these VNTRs is quite high at least for the WSSV strains, hosts and number of passages examined and that the VNTR sequences thus represent useful genetic markers for studying WSSV epidemiology.

  19. Horizontal transmission dynamics of White spot syndrome virus by cohabitation trials in juvenile Penaeus monodon and P. vannamei.

    PubMed

    Tuyen, N X; Verreth, J; Vlak, J M; de Jong, M C M

    2014-11-01

    White spot syndrome virus (WSSV), a rod-shaped double-stranded DNA virus, is an infectious agent causing fatal disease in shrimp farming around the globe. Within shrimp populations WSSV is transmitted very fast, however, the modes and dynamics of transmission of this virus are not well understood. In the current study the dynamics of disease transmission of WSSV were investigated in small, closed populations of Penaeus monodon and Penaeus vannamei. Pair cohabitation experiments using PCR as a readout for virus infection were used to estimate transmission parameters for WSSV in these two species. The mortality rate of contact-infected shrimp in P. monodon was higher than the rate in P. vannamei. The transmission rate parameters for WSSV were not different between the two species. The relative contribution of direct and indirect transmission rates of WSSV differed between the two species. For P. vannamei the direct contact transmission rate of WSSV was significantly lower than the indirect environmental transmission rate, but for P. monodon, the opposite was found. The reproduction ratio R0 for WSSV for these two species of shrimp was estimated to be above one: 2.07 (95%CI 1.53, 2.79) for P. monodon and 1.51 (95%CI 1.12, 2.03) for P. vannamei. The difference in R0 between the two species is due to a lower host mortality and hence a longer infectious period of WSSV in P. monodon.

  20. Comparative microarray profile of the hepatopancreas in the response of "Huanghai No. 2" Fenneropenaeus chinensis to white spot syndrome virus.

    PubMed

    Shi, Xiaoli; Kong, Jie; Meng, Xianhong; Luan, Sheng; Luo, Kun; Cao, Baoxiang; Liu, Ning; Lu, Xia; Deng, Kangyu; Cao, Jiawang; Zhang, Yingxue; Zhang, Hengheng; Li, Xupeng

    2016-11-01

    White spot syndrome virus (WSSV) infects all shrimp species and is the greatest detriment to shrimp culture. To better understand the mechanism of molecular responses to WSSV infection in "Huanghai No. 2" Fenneropenaeus chinensis, a microarray technique was used. Microarray gene expression profiling of 59,137 unigenes identified Differentially Expressed Genes (DEGs) both in live and moribund shrimp at early, peak and late phases. In live shrimp, 1307, 1479 and 1539 DEGs were obtained in the early, peak and late phase, respectively. Meanwhile, 1536, 2181 and 1591 DEGs were obtained in moribund shrimp. Twenty known annotation genes are uniquely expressed in the late phase of live shrimp, including adhesion regulating molecule 1, arginine kinase, BUD31 homolog, and QM. Compared to WSSV-susceptible shrimp, 75 known annotation genes are uniquely expressed in WSSV-resistant shrimp, including arginine kinase, BUD31 homolog, clottable protein 2, caspase 2, cathepsin C, calnexin, HMGBb, Histone 3, and selenoprotein M. The gene expression patterns of the infected shrimp were altered by WSSV infection. To further confirm the expression of differentially expressed genes, real-time RT-PCR was performed to test six randomly selected genes. The data will provide valuable information to understand the immune mechanism of shrimp's response to WSSV.

  1. The first detection of white spot syndrome virus in naturally infected cultured Chinese mitten crabs, Eriocheir sinensis in China.

    PubMed

    Ding, Zhengfeng; Yao, Yufeng; Zhang, Fengxiang; Wan, Jinjuan; Sun, Mengling; Liu, Hongyan; Zhou, Gang; Tang, Jianqing; Pan, Jianlin; Xue, Hui; Zhao, Ziming

    2015-08-01

    An epidemic with a high mortality rate (80-100%) recently occurred in the cultured Chinese mitten crab, Eriocheir sinensis, which is a very important economic crustacean species in China. Using negative stain, histopathology and nested PCR supplemented by sequencing we identified white spot syndrome virus (WSSV) in these crabs. Challenge experiments revealed that the disease was caused by WSSV and confirmed the crab's susceptibility to this virus, which was consistent with previous laboratory-based studies. A cumulative mortality of 100% was observed within 10 days post WSSV injection. This is the first report of WSSV-associated disease outbreaks in the Chinese mitten crab, which is normally reported as an important penaeid-shrimp viral pathogen. Furthermore, this is only the second report to describe a significant pathogen in pond-cultured E. sinensis. These results will enhance the early diagnosis of WSSV in the crab farms and help in monitoring efforts directed at determining the prevalence of the virus in E. sinensis.

  2. Envelope Proteins of White Spot Syndrome Virus (WSSV) Interact with Litopenaeus vannamei Peritrophin-Like Protein (LvPT).

    PubMed

    Xie, Shijun; Zhang, Xiaojun; Zhang, Jiquan; Li, Fuhua; Xiang, Jianhai

    2015-01-01

    White spot syndrome virus (WSSV) is a major pathogen in shrimp cultures. The interactions between viral proteins and their receptors on the surface of cells in a frontier target tissue are crucial for triggering an infection. In this study, a yeast two-hybrid (Y2H) library was constructed using cDNA obtained from the stomach and gut of Litopenaeus vannamei, to ascertain the role of envelope proteins in WSSV infection. For this purpose, VP37 was used as the bait in the Y2H library screening. Forty positive clones were detected after screening. The positive clones were analyzed and discriminated, and two clones belonging to the peritrophin family were subsequently confirmed as genuine positive clones. Sequence analysis revealed that both clones could be considered as the same gene, LV-peritrophin (LvPT). Co-immunoprecipitation confirmed the interaction between LvPT and VP37. Further studies in the Y2H system revealed that LvPT could also interact with other WSSV envelope proteins such as VP32, VP38A, VP39B, and VP41A. The distribution of LvPT in tissues revealed that LvPT was mainly expressed in the stomach than in other tissues. In addition, LvPT was found to be a secretory protein, and its chitin-binding ability was also confirmed.

  3. An Elegant Analysis of White Spot Syndrome Virus Using a Graphene Oxide/Methylene Blue based Electrochemical Immunosensor Platform

    PubMed Central

    Natarajan, Anusha; Devi, K. S. Shalini; Raja, Sudhakaran; Senthil Kumar, Annamalai

    2017-01-01

    White spot syndrome virus (WSSV) is a major devastating virus in aquaculture industry. A sensitive and selective diagnostic method for WSSV is a pressing need for the early detection and protection of the aquaculture farms. Herein, we first report, a simple electrochemical immunosensor based on methylene blue dye (MB) immobilized graphene oxide modified glassy carbon electrode (GCE/GO@MB) for selective, quick (35 ± 5 mins) and raw sample analysis of WSSV. The immunosensor was prepared by sequential modification of primary antibody, blocking agent (bovine serum album), antigen (as vp28 protein), secondary antibody coupled with horseradish peroxidase (Ab2-HRP) on the GCE/GO@MB. The modified electrode showed a well-defined redox peak at an equilibrium potential (E1/2), −0.4 V vs Ag/AgCl and mediated H2O2 reduction reaction without any false positive result and dissolved oxygen interferences in pH 7 phosphate buffer solution. Under an optimal condition, constructed calibration plot was linear in a range of 1.36 × 10−3 to 1.36 × 107 copies μL−1 of vp28. It is about four orders higher sensitive than that of the values observed with polymerase chain reaction (PCR) and western blot based WSSV detection techniques. Direct electrochemical immunosensing of WSSV in raw tissue samples were successfully demonstrated as a real sample system. PMID:28393929

  4. Label Free Detection of White Spot Syndrome Virus Using Lead Magnesium Niobate-Lead Titanate Piezoelectric Microcantilever Sensors

    PubMed Central

    Capobianco, Joseph; Shih, Wei-Heng; Leu, Jiann-Horng; Lo, Grace Chu-Fang; Shih, Wan Y.

    2011-01-01

    We have investigated rapid, label free detection of white spot syndrome virus (WSSV) using the first longitudinal extension resonance peak of five lead-magnesium niobate-lead titanate (PMN-PT) piezoelectric microcantilever sensors (PEMS) 1050-700 μm long and 850-485 μm wide constructed from 8 μm thick PMN-PT freestanding films. The PMN-PT PEMS were encapsulated with a 3-mercaptopropltrimethoxysilane (MPS) insulation layer and further coated with anti-VP28 and anti-VP664 antibodies to target the WSSV virions and nucleocapsids, respectively. By inserting the antibody-coated PEMS in a flowing virion or nucleocapsid suspension, label-free detection of the virions and nucleocapsids were respectively achieved by monitoring the PEMS resonance frequency shift. We showed that positive label-free detection of both the virion and the nucleocapsid could be achieved at a concentration of 100 virions (nucleocapsids)/ml or 10 virions (nucleocapsids)/100μl, comparable to the detection sensitivity of polymerase chain reaction (PCR). However, in contrast to PCR, PEMS detection was label-free, in-situ and rapid (less than 30 min), potentially requiring minimal or no sample preparation. PMID:20863681

  5. Highly Sensitive Detection of Low-Abundance White Spot Syndrome Virus by a Pre-Amplification PCR Method.

    PubMed

    Pan, Xiaoming; Zhang, Yanfang; Sha, Xuejiao; Wang, Jing; Li, Jing; Dong, Ping; Liang, Xingguo

    2017-03-28

    White spot syndrome virus (WSSV) is a major threat to the shrimp farming industry and so far there is no effective therapy for it, and thus early diagnostic of WSSV is of great importance. However, at the early stage of infection, the extremely low-abundance of WSSV DNA challenges the detection sensitivity and accuracy of PCR. To effectively detect low-abundance WSSV, here we developed a pre-amplification PCR (pre-amp PCR) method to amplify trace amounts of WSSV DNA from massive background genomic DNA. Combining with normal specific PCR, 10 copies of target WSSV genes were detected from ~10(10) magnitude of backgrounds. In particular, multiple target genes were able to be balanced amplified with similar efficiency due to the usage of the universal primer. The efficiency of the pre-amp PCR was validated by nested-PCR and quantitative PCR, and pre-amp PCR showed higher efficiency than nested-PCR when multiple targets were detected. The developed method is particularly suitable for the super early diagnosis of WSSV, and has potential to be applied in other low-abundance sample detection cases.

  6. Virulence and genotypes of white spot syndrome virus infecting Pacific white shrimp Litopenaeus vannamei in north-western Mexico.

    PubMed

    Ramos-Paredes, J; Grijalva-Chon, J M; Ibarra-Gámez, J C

    2017-03-01

    White spot syndrome virus (WSSV) has caused substantial global economic impact on aquaculture, and it has been determined that strains can vary in virulence. In this study, the effect of viral load was evaluated by infecting Litopenaeus vannamei with 10-fold serial dilution of tissue infected with strain WSSV Mx-H, and the virulence of four WSSV strains from north-western Mexico was assessed along with their variable number of tandem repeat (VNTR) genotypes in ORF75, ORF94 and ORF125. The LD50 of the Mx-H strain was a dilution dose of 10(-7.5) ; the mortality titre was 10(9.2) LD50 per gram. In shrimp injected with 10(2.5) to 10(6.5) LD50 , no significant virulence differences were evident. Using mortality data, the four WSSV strains grouped into three virulence levels. The Mx-F strain (intermediate virulence) and the Mx-C strain (high virulence) showed more genetic differences than those observed between the Mx-G (low-virulence) and Mx-H (high-virulence) strains, in ORF94 and ORF125. The application of high-viral-load inocula proved useful in determining the different virulence phenotypes of the WSSV strains from the Eastern Pacific.

  7. A white spot syndrome virus microRNA promotes the virus infection by targeting the host STAT

    PubMed Central

    Ren, Qian; Huang, Ying; He, Yaodong; Wang, Wen; Zhang, Xiaobo

    2015-01-01

    JAK/STAT pathway plays an important role in invertebrates during virus infection. However the microRNA (miRNA)-mediated regulation of JAK/STAT is not intensively investigated. Viral miRNAs, encoded by virus genome, have emerged as important regulators in the virus-host interactions. In this study, a WSSV (white spot syndrome virus)-encoded miRNA (WSSV-miR-22) was characterized in shrimp during virus infection. The results showed that the viral miRNA could promote WSSV infection in shrimp by targeting the host STAT gene. When the expression of JAK or STAT was knocked down by sequence-specific siRNA, the WSSV copies in shrimp were significantly increased, indicating that the JAK/STAT played positive roles in the antiviral immunity of shrimp. The further findings revealed that TEP1 and TEP2 were the effectors of JAK-STAT signaling pathway. The silencing of TEP1 or TEP2 led to an increase of WSSV copies in shrimp, showing TEP1 and TEP2 were involved in the shrimp immune response against virus infection. Therefore our study presented a novel viral miRNA-mediated JAK/STAT-TEP1/TEP2 signaling pathway in virus infection. PMID:26671453

  8. An Investigation into Occasional White Spot Syndrome Virus Outbreak in Traditional Paddy Cum Prawn Fields in India

    PubMed Central

    Selvam, Deborah Gnana; Mujeeb Rahiman, K. M.; Mohamed Hatha, A. A.

    2012-01-01

    A yearlong (September 2009–August 2010) study was undertaken to find out possible reasons for occasional occurrence of White Spot Syndrome Virus (WSSV) outbreak in the traditional prawn farms adjoining Cochin backwaters. Physicochemical and bacteriological parameters of water and sediment from feeder canal and four shrimp farms were monitored on a fortnightly basis. The physicochemical parameters showed variation during the two production cycles and between the farms studied. Dissolved oxygen (DO) content of water from feeder canal showed low oxygen levels (as low as 0.8 mg/L) throughout the study period. There was no disease outbreak in the perennial ponds. Poor water exchange coupled with nutrient loading from adjacent houses resulted in phytoplankton bloom in shallow seasonal ponds which led to hypoxic conditions in early morning and supersaturation of DO in the afternoon besides considerably high alkaline pH. Ammonia levels were found to be very high in these ponds. WSSV outbreak was encountered twice during the study leading to mass mortalities in the seasonal ponds. The hypoxia and high ammonia content in water and abrupt fluctuations in temperature, salinity and pH might lead to considerable stress in the shrimps triggering WSSV infection in these traditional ponds. PMID:22593673

  9. An investigation into occasional White Spot Syndrome Virus outbreak in traditional paddy cum prawn fields in India.

    PubMed

    Selvam, Deborah Gnana; Mujeeb Rahiman, K M; Mohamed Hatha, A A

    2012-01-01

    A yearlong (September 2009-August 2010) study was undertaken to find out possible reasons for occasional occurrence of White Spot Syndrome Virus (WSSV) outbreak in the traditional prawn farms adjoining Cochin backwaters. Physicochemical and bacteriological parameters of water and sediment from feeder canal and four shrimp farms were monitored on a fortnightly basis. The physicochemical parameters showed variation during the two production cycles and between the farms studied. Dissolved oxygen (DO) content of water from feeder canal showed low oxygen levels (as low as 0.8 mg/L) throughout the study period. There was no disease outbreak in the perennial ponds. Poor water exchange coupled with nutrient loading from adjacent houses resulted in phytoplankton bloom in shallow seasonal ponds which led to hypoxic conditions in early morning and supersaturation of DO in the afternoon besides considerably high alkaline pH. Ammonia levels were found to be very high in these ponds. WSSV outbreak was encountered twice during the study leading to mass mortalities in the seasonal ponds. The hypoxia and high ammonia content in water and abrupt fluctuations in temperature, salinity and pH might lead to considerable stress in the shrimps triggering WSSV infection in these traditional ponds.

  10. Protein profiling in the gut of Penaeus monodon gavaged with oral WSSV-vaccines and live white spot syndrome virus.

    PubMed

    Kulkarni, Amod D; Kiron, Viswanath; Rombout, Jan H W M; Brinchmann, Monica F; Fernandes, Jorge M O; Sudheer, Naduvilamuriparampu S; Singh, Bright I S

    2014-07-01

    White spot syndrome virus (WSSV) is a pathogen that causes considerable mortality of the farmed shrimp, Penaeus monodon. Candidate 'vaccines', WSSV envelope protein VP28 and formalin-inactivated WSSV, can provide short-lived protection against the virus. In this study, P. monodon was orally intubated with the aforementioned vaccine candidates, and protein expression in the gut of immunised shrimps was profiled. The alterations in protein profiles in shrimps infected orally with live-WSSV were also examined. Seventeen of the identified proteins in the vaccine and WSSV-intubated shrimps varied significantly compared to those in the control shrimps. These proteins, classified under exoskeletal, cytoskeletal, immune-related, intracellular organelle part, intracellular calcium-binding or energy metabolism, are thought to directly or indirectly affect shrimp's immunity. The changes in the expression levels of crustacyanin, serine proteases, myosin light chain, and ER protein 57 observed in orally vaccinated shrimp may probably be linked to immunoprotective responses. On the other hand, altered expression of proteins linked to exoskeleton, calcium regulation and energy metabolism in WSSV-intubated shrimps is likely to symbolise disturbances in calcium homeostasis and energy metabolism.

  11. Risk factors associated with white spot syndrome virus infection in a Vietnamese rice-shrimp farming system.

    PubMed

    Corsin, F; Turnbull, J F; Hao, N V; Mohan, C V; Phi, T T; Phuoc, L H; Tinh, N T; Morgan, K L

    2001-10-29

    White spot disease (WSD) is a pandemic disease caused by a virus commonly known as white spot syndrome virus (WSSV). Several risk factors for WSD outbreaks have been suggested. However, there have been very few studies to identify risk factors for WSD outbreaks in culture systems. This paper presents and discusses the risk factors for WSSV infection identified during a longitudinal observational study conducted in a Vietnamese rice-shrimp farming system. A total of 158 variables were measured comprising location, features of the pond, management practices, pond bottom quality, shrimp health and other animals in the pond. At the end of the study period WSSV was detected in 15 of the 24 ponds followed through the production cycle (62.5%). One hundred and thirty-nine variables were used in univariate analyses. All the variables with a p-value < or = 0.10 were used in unconditional logistic regression in a forward stepwise model. An effect of location was identified in both univariate and multivariate analyses showing that ponds located in the eastern portion of the study site, closer to the sea, were more likely to test positive for WSSV by 1-step PCR at harvest. Ponds with shrimp of a smaller average size 1 mo after stocking tended to be positive for WSSV at the end of the production cycle. Average weight at 1 mo was also highlighted in multivariate analyses when considered as either a risk factor or an outcome. Other risk factors identified in univariate analyses were earlier date of stocking and use of commercial feed. A number of variables also appeared to be associated with a reduced risk of WSSV at harvest including the presence of dead post larvae in the batch sampled at stocking, presence of Hemigrapsus spp. crabs during the first month of production, feeding vitamin premix or legumes, presence of high numbers of shrimp with bacterial infection and the presence of larger mud crabs or gobies at harvest. No associations were detected with WSSV at harvest and

  12. Prohibitin Interacts with Envelope Proteins of White Spot Syndrome Virus and Prevents Infection in the Red Swamp Crayfish, Procambarus clarkii

    PubMed Central

    Lan, Jiang-Feng; Li, Xin-Cang; Sun, Jie-Jie; Gong, Jing; Wang, Xian-Wei; Shi, Xiu-Zhen; Shi, Li-Jie; Weng, Yu-Ding; Zhao, Xiao-Fan

    2013-01-01

    Prohibitins (PHBs) are ubiquitously expressed conserved proteins in eukaryotes that are associated with apoptosis, cancer formation, aging, stress responses, cell proliferation, and immune regulation. However, the function of PHBs in crustacean immunity remains largely unknown. In the present study, we identified a PHB in Procambarus clarkii red swamp crayfish, which was designated PcPHB1. PcPHB1 was widely distributed in several tissues, and its expression was significantly upregulated by white spot syndrome virus (WSSV) challenge at the mRNA level and the protein level. These observations prompted us to investigate the role of PcPHB1 in the crayfish antiviral response. Recombinant PcPHB1 (rPcPHB1) significantly reduced the amount of WSSV in crayfish and the mortality of WSSV-infected crayfish. The quantity of WSSV in PcPHB1 knockdown crayfish was increased compared with that in the controls. The effects of RNA silencing were rescued by rPcPHB1 reinjection. We further confirmed the interaction of PcPHB1 with the WSSV envelope proteins VP28, VP26, and VP24 using pulldown and far-Western overlay assays. Finally, we observed that the colloidal gold-labeled PcPHB1 was located on the outer surface of the WSSV, which suggests that PcPHB1 specifically binds to the envelope proteins of WSSV. VP28, VP26, and VP24 are structural envelope proteins and are essential for attachment and entry into crayfish cells. Therefore, PcPHB1 exerts its anti-WSSV effect by binding to VP28, VP26, and VP24, preventing viral infection. This study is the first report on the antiviral function of PHB in the innate immune system of crustaceans. PMID:24049173

  13. Studies of the viral binding proteins of shrimp BP53, a receptor of white spot syndrome virus.

    PubMed

    Li, Chen; Gao, Xiao-Xiao; Huang, Jie; Liang, Yan

    2016-02-01

    The specific binding between viral attachment proteins (VAPs) of a virus and its cellular receptors on host cells mediates virus entry into host cells, which triggers subsequent viral infections. Previous studies indicate that F1 ATP synthase β subunit (named BP53), is found on the surface of shrimp cells and involved in white spot syndrome virus (WSSV) infection by functioning as a potential viral receptor. Herein, in a far-western blotting assay, three WSSV proteins with molecular weights of 28 kDa, 37 kDa, and >50 kDa were found to interact with BP53. The 28 kDa and 37 kDa proteins were identified as the envelope protein VP28 and VP37 of WSSV respectively, which could be recognized by the polyclonal antibodies. Enzyme-linked immunosorbent binding assays revealed that VP37 contributed to almost 80% of the binding capability for BP53 compared with the same amount of total WSSV protein. The relationship between BP53 and its complementary interacting protein, VP37, was visualized using a co-localization assay. Bound VP37 on the cell surface co-localized with BP53 and shared a similar subcellular location on the outer surface of shrimp cells. Pearson's correlation coefficients reached to 0.67 ± 0.05 and the Mander's overlap coefficients reached 0.70 ± 0.05, which indicated a strong relationship between the localization of BP53 and bound rVP37. This provides evidence for an interaction between BP53 and VP37 obtained at the molecular and cellular levels, supporting the hypothesis that BP53 serves as a receptor for WSSV by binding to VP37. The identification of the viral binding proteins of shrimp BP53 is helpful for better understanding the pathogenic mechanisms of WSSV to infect shrimp at the cellular level.

  14. Transcript Analysis of White spot syndrome virus Latency and Phagocytosis Activating Protein Genes in Infected Shrimp (Penaeus monodon).

    PubMed

    Shekhar, M S; Dillikumar, M; Vinaya Kumar, K; Gopikrishna, G; Rajesh, S; Kiruthika, J; Ponniah, A G

    2012-12-01

    Viral latency has been recently observed to be associated with White spot syndrome virus (WSSV) infection in shrimp. In the present study, shrimp samples (Penaeus monodon) surviving WSSV infection were examined for presence of WSSV in latent phase. Virus latency was observed in shrimp which were either experimentally challenged with WSSV and survived the infection or those which survived the natural infection. Three viral transcripts (ORFs 427, 151, 366) associated with latency were analyzed by real-time PCR. The shrimp surviving the natural WSSV infection on estimation with RT-PCR were found to have low grade of WSSV infection (less than 56 copies of WSSV). All the shrimp samples were RT-PCR negative for structural protein genes of WSSV, VP24 and VP28, indicating that these samples were harboring latent phase virus. RT-PCR of all the shrimp samples which survived WSSV infection revealed amplification of phagocytosis activating protein (PAP) gene (435 bp) with higher gene expression levels in experimentally challenged shrimp when compared to naturally infected shrimp. The expression of PAP in WSSV infected shrimp samples indicates its possible role in host response for resistance against WSSV infection. PAP was cloned and expressed as recombinant protein for protection studies. Shrimp were injected with three doses (5, 15 and 20 μg g(-1) body weight) of recombinant PAP. Relative percent survival of 10 % was observed in shrimp immunized with the dose of 15 μg g(-1) body weight of recombinant PAP. The expression of both WSSV latency associated and PAP genes obtained from shrimp surviving the WSSV infection, indicates the possible role of these genes in host-pathogen interaction.

  15. Prohibitin Interacts with envelope proteins of white spot syndrome virus and prevents infection in the red swamp crayfish, Procambarus clarkii.

    PubMed

    Lan, Jiang-Feng; Li, Xin-Cang; Sun, Jie-Jie; Gong, Jing; Wang, Xian-Wei; Shi, Xiu-Zhen; Shi, Li-Jie; Weng, Yu-Ding; Zhao, Xiao-Fan; Wang, Jin-Xing

    2013-12-01

    Prohibitins (PHBs) are ubiquitously expressed conserved proteins in eukaryotes that are associated with apoptosis, cancer formation, aging, stress responses, cell proliferation, and immune regulation. However, the function of PHBs in crustacean immunity remains largely unknown. In the present study, we identified a PHB in Procambarus clarkii red swamp crayfish, which was designated PcPHB1. PcPHB1 was widely distributed in several tissues, and its expression was significantly upregulated by white spot syndrome virus (WSSV) challenge at the mRNA level and the protein level. These observations prompted us to investigate the role of PcPHB1 in the crayfish antiviral response. Recombinant PcPHB1 (rPcPHB1) significantly reduced the amount of WSSV in crayfish and the mortality of WSSV-infected crayfish. The quantity of WSSV in PcPHB1 knockdown crayfish was increased compared with that in the controls. The effects of RNA silencing were rescued by rPcPHB1 reinjection. We further confirmed the interaction of PcPHB1 with the WSSV envelope proteins VP28, VP26, and VP24 using pulldown and far-Western overlay assays. Finally, we observed that the colloidal gold-labeled PcPHB1 was located on the outer surface of the WSSV, which suggests that PcPHB1 specifically binds to the envelope proteins of WSSV. VP28, VP26, and VP24 are structural envelope proteins and are essential for attachment and entry into crayfish cells. Therefore, PcPHB1 exerts its anti-WSSV effect by binding to VP28, VP26, and VP24, preventing viral infection. This study is the first report on the antiviral function of PHB in the innate immune system of crustaceans.

  16. Virus replication cycle of white spot syndrome virus in secondary cell cultures from the lymphoid organ of Litopenaeus vannamei.

    PubMed

    Li, Wenfeng; Desmarets, Lowiese M B; De Gryse, Gaëtan M A; Theuns, Sebastiaan; Van Tuan, Vo; Van Thuong, Khuong; Bossier, Peter; Nauwynck, Hans J

    2015-09-01

    The replication cycle of white spot syndrome virus (WSSV) was investigated in secondary cell cultures from the lymphoid organ of Litopenaeus vannamei. The secondary cells formed a confluent monolayer at 24 h post-reseeding, and this monolayer could be maintained for 10 days with a viability of 90 %. Binding of WSSV to cells reached a maximum (73 ± 3 % of cells and 4.84 ± 0.2 virus particles per virus-binding cell) at 120 min at 4 °C. WSSV entered cells by endocytosis. The co-localization of WSSV and early endosomes was observed starting from 30 min post-inoculation (p.i.). Double indirect immunofluorescence staining showed that all cell-bound WSSV particles entered these cells in the period between 0 and 60 min p.i. and that the uncoating of WSSV occurred in the same period. After 1 h inoculation at 27 °C, the WSSV nucleocapsid protein VP664 and envelope protein VP28 started to be synthesized in the cytoplasm from 1 and 3 h p.i., and were transported into nuclei from 3 and 6 h p.i., respectively. The percentage of cells that were VP664- and VP28-positive in their nuclei peaked (50 ± 4 %) at 12 h p.i. Quantitative PCR showed that WSSV DNA started to be synthesized from 6 h p.i. In vivo titration of the supernatants showed that the progeny WSSV were released from 12 h p.i. and peaked at 18 h p.i. In conclusion, the secondary cell cultures from the lymphoid organ were proven to be ideal for examination of the replication cycle of WSSV.

  17. Isolation and expression analysis of an MAPKK gene from Fenneropenaeus chinensis in response to white spot syndrome virus infection.

    PubMed

    Li, Xupeng; Kong, Jie; Meng, Xianhong; Luo, Kun; Luan, Sheng; Cao, Baoxiang; Liu, Ning

    2016-08-01

    Mitogen-activated kinase kinase (MAPKK) is an important gene involved in the host-virus interaction process. To obtain a better understanding of MAPKK in the interaction process between the Chinese shrimp Fenneropenaeus chinensis and white spot syndrome virus (WSSV), we cloned the sequence of an MAPKK cDNA from F. chinensis (FcMAPKK) and investigated the effect of FcMAPKK on WSSV infection. The results showed that the FcMAPKK gene contained a 1227 bp open reading frame (ORF), which encoded a highly conserved protein with a serine/threonine protein kinase catalytic (S_TKc) domain. The deduced amino acid sequence of FcMAPKK shared identities between 11.9 and 92.6% with MAPKKs from vertebrate, invertebrate, plant and fungus species. The FcMAPKK was expressed in all the examined tissues in the normal F. chinensis. FcMAPKK expression level was highest in the hepatopancreas where it was approximately 2.6-fold the expression level in the gill, and lowest in the muscle where it was approximately 0.3-fold the expression level in the hepatopancreas. The FcMAPKK expression levels in the muscle, gill, and hepatopancreas were all changed post WSSV challenge. The FcMAPKK expression was significantly (P < 0.01) up-regulated in the muscle of F. chinensis at 48 h post WSSV infection. The WSSV began to replicate quickly in the normal F. chinensis at 48 h post infection, while the WSSV replication in the U0126-treated F. chinensis could be significantly (P < 0.05) inhibited. The results suggested that FcMAPKK might be involved in the WSSV infection process, and hijacking of FcMAPKK might be required for WSSV replication in F. chinensis.

  18. Evaluation of white spot syndrome virus variable DNA loci as molecular markers of virus spread at intermediate spatiotemporal scales.

    PubMed

    Dieu, Bui Thi Minh; Marks, Hendrik; Zwart, Mark P; Vlak, Just M

    2010-05-01

    Variable genomic loci have been employed in a number of molecular epidemiology studies of white spot syndrome virus (WSSV), but it is unknown which loci are suitable molecular markers for determining WSSV spread on different spatiotemporal scales. Although previous work suggests that multiple introductions of WSSV occurred in central Vietnam, it is largely uncertain how WSSV was introduced and subsequently spread. Here, we evaluate five variable WSSV DNA loci as markers of virus spread on an intermediate (i.e. regional) scale, and develop a detailed and statistically supported model for the spread of WSSV. The genotypes of 17 WSSV isolates from along the coast of Vietnam--nine of which were newly characterized in this study--were analysed to obtain sufficient samples on an intermediate scale and to allow statistical analysis. Only the ORF23/24 variable region is an appropriate marker on this scale, as geographically proximate isolates show similar deletion sizes. The ORF14/15 variable region and variable-number tandem repeat (VNTR) loci are not useful as markers on this scale. ORF14/15 may be suitable for studying larger spatiotemporal scales, whereas VNTR loci are probably suitable for smaller scales. For ORF23/24, there is a clear pattern in the spatial distribution of WSSV: the smallest genomic deletions are found in central Vietnam, and larger deletions are found in the south and the north. WSSV genomic deletions tend to increase over time with virus spread in cultured shrimp, and our data are therefore congruent with the hypothesis that WSSV was introduced in central Vietnam and then radiated out.

  19. Immune gene expression profile of Penaeus monodon in response to marine yeast glucan application and white spot syndrome virus challenge.

    PubMed

    Wilson, Wilsy; Lowman, Douglas; Antony, Swapna P; Puthumana, Jayesh; Bright Singh, I S; Philip, Rosamma

    2015-04-01

    Immunostimulant potential of eight marine yeast glucans (YG) from Candida parapsilosis R20, Hortaea werneckii R23, Candida spencermartinsiae R28, Candida haemulonii R63, Candida oceani R89, Debaryomyces fabryi R100, Debaryomyces nepalensis R305 and Meyerozyma guilliermondii R340 were tested against WSSV challenge in Penaeus monodon post larvae (PL). Structural characterization of these marine yeast glucans by proton nuclear magnetic resonance (NMR) indicated structures containing (1-6)-branched (1-3)-β-D-glucan. PL were fed 0.2% glucan incorporated diet once in seven days for a period of 45 days and the animals were challenged with white spot syndrome virus (WSSV). The immunostimulatory activity of yeast glucans were assessed pre- and post-challenge WSSV by analysing the expression profile of six antimicrobial peptide (AMP) genes viz., anti-lipopolysaccharide factor (ALF), crustin-1, crustin-2, crustin-3, penaeidin-3 and penaeidin-5 and 13 immune genes viz., alpha-2-macroglobulin (α-2-M), astakine, caspase, catalase, glutathione peroxidase, glutathione-s-transferase, haemocyanin, peroxinectin, pmCathepsinC, prophenol oxidase (proPO), Rab-7, superoxide dismutase and transglutaminase. Expression of seven WSSV genes viz., DNA polymerase, endonuclease, protein kinase, immediate early gene, latency related gene, thymidine kinase and VP28 were also analysed to detect the presence and intensity of viral infection in the experimental animals post-challenge. The study revealed that yeast glucans (YG) do possess immunostimulatory activity against WSSV and also supported higher survival (40-70 %) post-challenge WSSV. Among the various glucans tested, YG23 showed maximum survival (70.27%), followed by YG20 (66.66%), YG28 (60.97%), YG89 (58.53%), YG100 (54.05%), YG63 (48.64%), YG305 (45.7%) and YG340 (43.24%).

  20. Interaction of the Small GTPase Cdc42 with Arginine Kinase Restricts White Spot Syndrome Virus in Shrimp.

    PubMed

    Xu, Ji-Dong; Jiang, Hai-Shan; Wei, Tian-Di; Zhang, Ke-Yi; Wang, Xian-Wei; Zhao, Xiao-Fan; Wang, Jin-Xing

    2017-03-01

    Many types of small GTPases are widely expressed in eukaryotes and have different functions. As a crucial member of the Rho GTPase family, Cdc42 serves a number of functions, such as regulating cell growth, migration, and cell movement. Several RNA viruses employ Cdc42-hijacking tactics in their target cell entry processes. However, the function of Cdc42 in shrimp antiviral immunity is not clear. In this study, we identified a Cdc42 protein in the kuruma shrimp (Marsupenaeus japonicus) and named it MjCdc42. MjCdc42 was upregulated in shrimp challenged by white spot syndrome virus (WSSV). The knockdown of MjCdc42 and injection of Cdc42 inhibitors increased the proliferation of WSSV. Further experiments determined that MjCdc42 interacted with an arginine kinase (MjAK). By analyzing the binding activity and enzyme activity of MjAK and its mutant, ΔMjAK, we found that MjAK could enhance the replication of WSSV in shrimp. MjAK interacted with the envelope protein VP26 of WSSV. An inhibitor of AK activity, quercetin, could impair the function of MjAK in WSSV replication. Further study demonstrated that the binding of MjCdc42 and MjAK depends on Cys(271) of MjAK and suppresses the WSSV replication-promoting effect of MjAK. By interacting with the active site of MjAK and suppressing its enzyme activity, MjCdc42 inhibits WSSV replication in shrimp. Our results demonstrate a new function of Cdc42 in the cellular defense against viral infection in addition to the regulation of actin and phagocytosis, which has been reported in previous studies. IMPORTANCE The interaction of Cdc42 with arginine kinase plays a crucial role in the host defense against WSSV infection. This study identifies a new mechanism of Cdc42 in innate immunity and enriches the knowledge of the antiviral innate immunity of invertebrates.

  1. Injected phage-displayed-VP28 vaccine reduces shrimp Litopenaeus vannamei mortality by white spot syndrome virus infection.

    PubMed

    Solís-Lucero, G; Manoutcharian, K; Hernández-López, J; Ascencio, F

    2016-08-01

    White spot syndrome virus (WSSV) is the most important viral pathogen for the global shrimp industry causing mass mortalities with huge economic losses. Recombinant phages are capable of expressing foreign peptides on viral coat surface and act as antigenic peptide carriers bearing a phage-displayed vaccine. In this study, the full-length VP28 protein of WSSV, widely known as potential vaccine against infection in shrimp, was successfully cloned and expressed on M13 filamentous phage. The functionality and efficacy of this vaccine immunogen was demonstrated through immunoassay and in vivo challenge studies. In ELISA assay phage-displayed VP28 was bind to Litopenaeus vannamei immobilized hemocyte in contrast to wild-type M13 phage. Shrimps were injected with 2 × 10(10) cfu animal(-1) single dose of VP28-M13 and M13 once and 48 h later intramuscularly challenged with WSSV to test the efficacy of the vaccine against the infection. All dead challenged shrimps were PCR WSSV-positive. The accumulative mortality of the vaccinated and challenged shrimp groups was significantly lower (36.67%) than the unvaccinated group (66.67%). Individual phenoloxidase and superoxide dismutase activity was assayed on 8 and 48 h post-vaccination. No significant difference was found in those immunological parameters among groups at any sampled time evaluated. For the first time, phage display technology was used to express a recombinant vaccine for shrimp. The highest percentage of relative survival in vaccinated shrimp (RPS = 44.99%) suggest that the recombinant phage can be used successfully to display and deliver VP28 for farmed marine crustaceans.

  2. McCune Albright syndrome - association of fibrous dysplasia, café-au-lait skin spots and hyperthyroidism – case report

    PubMed Central

    RAUS, IULIAN; COROIU, ROXANA ELENA

    2016-01-01

    McCune–Albright syndrome is a rare sporadic disease characterized by bone fibrous dysplasia, café-au-lait skin spots and a variable association of hyperfunctional endocrine disorders. Fibrous dysplasia (FD), which can involve the craniofacial, axial, and appendicular skeleton, may range from an isolated, asymptomatic monostotic lesion to a severe disabling polyostotic disease involving the entire skeleton. A twenty-five-year old male patient presented to our clinic with recently developed heart palpitations. He had also been feeling pain in the right femur since he was younger, without any trauma history, leading to difficulties of ambulation and limping occasionally. His physical examination revealed café-au-lait spots with irregular borders and right testicular agenesis. Laboratory findings identified hyperthyroidism with hyperparathyroidism. Radiographs of the pelvis revealed multiple lytic lesions of the right femur and magnetic resonance imaging (MRI) characterized these lesions as specific to fibrous dysplasia of the bone, without any insufficiency fracture at this level. The association of café-au-lait skin spots with bone fibrous dysplasia, and hyperthyroidism in this patient suggested the diagnosis of McCune – Albright syndrome. PMID:27857528

  3. A prospective clinical trial to compare the performance of dried blood spots prenatal screening for Down's syndrome with conventional non-invasive testing technology.

    PubMed

    Hu, Huiying; Jiang, Yulin; Zhang, Minghui; Liu, Shanying; Hao, Na; Zhou, Jing; Liu, Juntao; Zhang, Xiaojin; Ma, Liangkun

    2017-03-01

    To evaluate, side by side, the efficiency of dried blood spots (DBSs) against serum screening for Down's syndrome, and then, to construct a two-tier strategy by topping up the fetal cell-free DNA (cfDNA) secondary screening over the high-risk women marked by the primary blood testing to build a practical screening tactic to identify fetal Down's syndrome. One thousand eight hundred and thirty-seven low-risk Chinese women, with singleton pregnancy, were enrolled for the study. Alpha-fetoprotein and free beta human chorionic gonadotropin were measured for the serum as well as for the parallel DBS samples. Partial high-risk pregnant women identified by primary blood testing (n = 38) were also subject to the secondary cfDNA screening. Diagnostic amniocentesis was utilized to confirm the screening results. The true positive rate for Down's syndrome detection was 100% for both blood screening methods; however, the false-positive rate was 3.0% for DBS and 4.0% for serum screening, respectively. DBS correlated well with serum screening on Down's syndrome detection. Three out of 38 primary high-risk women displayed chromosomal abnormalities by cfDNA analysis, which were confirmed by amniocentesis. Either the true detection rate or the false-positive rate for Down's syndrome between DBS and the serum test is comparable. In addition, blood primary screening aligned with secondary cfDNA analysis, a "before and after" two-tier screening strategy, can massively decrease the false-positive rate, which, then, dramatically reduces the demand for invasive diagnostic operation. Impact statement Children born with Down's syndrome display a wide range of mental and physical disability. Currently, there is no effective treatment to ease the burden and anxiety of the Down's syndrome family and the surrounding society. This study is to evaluate the efficiency of dried blood spots against serum screening for Down's syndrome and to construct a two-tier strategy by topping up the fetal

  4. Yeast Surface Display of Two Proteins Previously Shown to Be Protective Against White Spot Syndrome Virus (WSSV) in Shrimp.

    PubMed

    Ananphongmanee, Vorawit; Srisala, Jiraporn; Sritunyalucksana, Kallaya; Boonchird, Chuenchit

    2015-01-01

    Cell surface display using the yeasts Saccharomyces cerevisiae and Pichia pastoris has been extensively developed for application in bioindustrial processes. Due to the rigid structure of their cell walls, a number of proteins have been successfully displayed on their cell surfaces. It was previously reported that the viral binding protein Rab7 from the giant tiger shrimp Penaeus monodon (PmRab7) and its binding partner envelope protein VP28 of white spot syndrome virus (WSSV) could independently protect shrimp against WSSV infection. Thus, we aimed to display these two proteins independently on the cell surfaces of 2 yeast clones with the ultimate goal of using a mixture of the two clones as an orally deliverable, antiviral agent to protect shrimp against WSSV infection. PmRab7 and VP28 were modified by N-terminal tagging to the C-terminal half of S. cerevisiae α-agglutinin. DNA fragments, harboring fused-gene expression cassettes under control of an alcohol oxidase I (AOX1) promoter were constructed and used to transform the yeast cells. Immunofluorescence microscopy with antibodies specific to both proteins demonstrated that mutated PmRab7 (mPmRab7) and partial VP28 (pVP28) were localized on the cell surfaces of the respective clones, and fluorescence intensity for each was significantly higher than that of control cells by flow cytometry. Enzyme-linked immunosorbant assay (ELISA) using cells displaying mPmRab7 or pVP28 revealed that the binding of specific antibodies for each was dose-dependent, and could be saturated. In addition, the binding of mPmRab7-expressing cells with free VP28, and vice versa was dose dependent. Binding between the two surface-expressed proteins was confirmed by an assay showing agglutination between cells expressing complementary mPmRab7 and pVP28. In summary, our genetically engineered P. pastoris can display biologically active mPmRab7 and pVP28 and is now ready for evaluation of efficacy in protecting shrimp against WSSV by oral

  5. Impact of Vibrio parahaemolyticus and white spot syndrome virus (WSSV) co-infection on survival of penaeid shrimp Litopenaeus vannamei

    NASA Astrophysics Data System (ADS)

    Zhang, Xiaojing; Song, Xiaoling; Huang, Jie

    2016-11-01

    White spot syndrome virus (WSSV) is an important viral pathogen that infects farmed penaeid shrimp, and the threat of Vibrio parahaemolyticus infection to shrimp farming has become increasingly severe. Viral and bacterial cross or superimposed infections may induce higher shrimp mortality. We used a feeding method to infect Litopenaeus vannamei with WSSV and then injected a low dose of V. parahaemolyticus (WSSV+Vp), or we first infected L. vannamei with a low-dose injection of V. parahaemolyticus and then fed the shrimp WSSV to achieve viral infection (Vp+WSSV). The eff ect of V. parahaemolyticus and WSSV co-infection on survival of L. vannamei was evaluated by comparing cumulative mortality rates between experimental and control groups. We also spread L. vannamei hemolymph on thiosulfate citrate bile salt sucrose agar plates to determine the number of Vibrio, and the WSSV copy number in L. vannamei gills was determined using an absolute quantitative polymerase chain reaction (PCR) method. LvMyD88 and Lvakt gene expression levels were detected in gills of L. vannamei by real-time PCR to determine the cause of the diff erent mortality rates. Our results show that (1) the cumulative mortality rate of L. vannamei in the WSSV+Vp group reached 100% on day 10 after WSSV infection, whereas the cumulative mortality rate of L. vannamei in the Vp+WSSV group and the WSSV-alone control group approached 100% on days 11 and 13 of infection; (2) the number of Vibrio in the L. vannamei group infected with V. parahaemolyticus alone declined gradually, whereas the other groups showed significant increases in the numbers of Vibrio ( P<0.05); (3) the WSSV copy numbers in the gills of the WSSV+Vp, Vp+WSSV, and the WSSV-alone groups increased from 105 to 107 /mg tissue 72, 96, and 144 h after infection, respectively. These results suggest that V. parahaemolyticus infection accelerated proliferation of WSSV in L. vannamei and vice versa. The combined accelerated proliferation of both V

  6. Yeast Surface Display of Two Proteins Previously Shown to Be Protective Against White Spot Syndrome Virus (WSSV) in Shrimp

    PubMed Central

    Ananphongmanee, Vorawit; Srisala, Jiraporn; Sritunyalucksana, Kallaya; Boonchird, Chuenchit

    2015-01-01

    Cell surface display using the yeasts Saccharomyces cerevisiae and Pichia pastoris has been extensively developed for application in bioindustrial processes. Due to the rigid structure of their cell walls, a number of proteins have been successfully displayed on their cell surfaces. It was previously reported that the viral binding protein Rab7 from the giant tiger shrimp Penaeus monodon (PmRab7) and its binding partner envelope protein VP28 of white spot syndrome virus (WSSV) could independently protect shrimp against WSSV infection. Thus, we aimed to display these two proteins independently on the cell surfaces of 2 yeast clones with the ultimate goal of using a mixture of the two clones as an orally deliverable, antiviral agent to protect shrimp against WSSV infection. PmRab7 and VP28 were modified by N-terminal tagging to the C-terminal half of S. cerevisiae α-agglutinin. DNA fragments, harboring fused-gene expression cassettes under control of an alcohol oxidase I (AOX1) promoter were constructed and used to transform the yeast cells. Immunofluorescence microscopy with antibodies specific to both proteins demonstrated that mutated PmRab7 (mPmRab7) and partial VP28 (pVP28) were localized on the cell surfaces of the respective clones, and fluorescence intensity for each was significantly higher than that of control cells by flow cytometry. Enzyme-linked immunosorbant assay (ELISA) using cells displaying mPmRab7 or pVP28 revealed that the binding of specific antibodies for each was dose-dependent, and could be saturated. In addition, the binding of mPmRab7-expressing cells with free VP28, and vice versa was dose dependent. Binding between the two surface-expressed proteins was confirmed by an assay showing agglutination between cells expressing complementary mPmRab7 and pVP28. In summary, our genetically engineered P. pastoris can display biologically active mPmRab7 and pVP28 and is now ready for evaluation of efficacy in protecting shrimp against WSSV by oral

  7. Liver spots

    MedlinePlus

    Sun-induced skin changes - liver spots; Senile or solar lentigines; Skin spots - aging; Age spots ... Liver spots are changes in skin color that occur in older skin. The coloring may be due to aging, exposure to the sun or other sources of ...

  8. A pilot metabolic profiling study in hepatopancreas of Litopenaeus vannamei with white spot syndrome virus based on ¹H NMR spectroscopy.

    PubMed

    Liu, Peng-fei; Liu, Qing-hui; Wu, Yin; Jie, Huang

    2015-01-01

    White spot syndrome virus, which was a pathogen first found in 1992, had emerged globally affecting shrimp populations in aquaculture. Here, we comprehensively analyzed the metabolic changes of hepatopancreas from Litopenaeus vannamei which were infected with white spot syndrome virus by (1)H nuclear magnetic resonance (NMR). Through the NOESYPR1D spectrum combined with multi-variate pattern recognition analysis, including principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA) models, significantly metabolic changes were observed in WSSV-infected groups compared with the control groups. In the first 48 h, α-glucose and β-glucose were higher in the WSSV-infected group. Meanwhile, acetate, lactate, N-acetyl glycoprotein signals, lysine, tyrosine and lipid were significantly decreased in the WSSV-infected group. These results suggest that WSSV caused absorption inhibition of amino acids and disturbed protein metabolism as well as cell metabolism in favor of its replication. Our findings could also contribute to further understanding of disease mechanisms.

  9. Interaction between Kazal serine proteinase inhibitor SPIPm2 and viral protein WSV477 reduces the replication of white spot syndrome virus.

    PubMed

    Ponprateep, Sirikwan; Phiwsaiya, Kornsunee; Tassanakajon, Anchalee; Rimphanitchayakit, Vichien

    2013-09-01

    White spot syndrome (WSS) is a viral disease caused by white spot syndrome virus (WSSV) which leads to severe mortality in cultured penaeid shrimp. In response to WSSV infection in Penaeus monodon, a Kazal serine proteinase inhibitor SPIPm2, normally stored in the granules of granular and semi-granular hemocytes is up-regulated and found to deter the viral replication. By using yeast two-hybrid screening, we have identified a viral target protein, namely WSV477. Instead of being a proteinase, the WSV477 was reported to be a Cys2/Cys2-type zinc finger regulatory protein having ATP/GTP-binding activity. In vitro pull down assay confirmed the protein-protein interaction between rSPIPm2 and rWSV477. Confocal laser scanning microscopy demonstrated that the SPIPm2 and WSV477 were co-localized in the cytoplasm of shrimp hemocytes. Using RNA interference, the silencing of WSV477 resulted in down-regulated of viral late gene VP28, the same result obtained with SPIPm2. In this instance, the SPIPm2 does not function as proteinase inhibitor but inhibit the regulatory function of WSV477.

  10. Haemocyanin content of shrimp (Fenneropenaeus chinensis) associated with white spot syndrome virus and Vibrio harveyi infection process.

    PubMed

    Chang, Yanhong; Xing, Jing; Tang, Xiaoqian; Sheng, Xiuzhen; Zhan, Wenbin

    2016-01-01

    Haemocyanin (Hc) is frequently reported to vary significantly by physiological status and environmental stress in Crustaceans. In this paper, the shrimp Fenneropenaeus chinensis was infected with different concentrations of white spot syndrome virus (WSSV) and Vibrio harveyi. Then, the variation of Hc and total protein content of the haemolymph (TPCH) were investigated using the established double antibody sandwich enzyme linked immunosorbent assay (DAS-ELISA) and Coomassie brilliant blue method, respectively. The results showed that the Hc content peaked at 12 h post-infection (PI) in the 10(-2), 10(-4) and 10(-6) viral supernatant (VS) groups, and the maximum was 93.03 ± 2.55 mg ml(-1), 77.57 ± 6.02 mg ml(-1) and 70.25 ± 3.96 mg ml(-1), respectively. TPCH reached the maximum of 108.18 ± 1.36 mg ml(-1) and 103.49 ± 1.33 mg ml(-1) at 12 h PI in the 10(-2) and 10(-4) VS groups, respectively. The maximum was 96.94 ± 1.06 mg ml(-1) at 24 h PI in the 10(-6) VS group. In the V. harveyi infection groups, the Hc content reached a maximum of 87.97 ± 4.39 mg ml(-1) at 36 h PI in the 10(6) CFU ml(-1) group, 73.74 ± 4.38 mg ml(-1) and 72.47 ± 2.09 mg ml(-1) at 12 h PI in the 10(7) and 10(8) CFU ml(-1) groups, respectively. TPCH reached a maximum of 111.16 ± 0.86 mg ml(-1) at 36 h PI in the 10(6) CFU ml(-1) group, 100.41 ± 0.51 mg ml(-1) and 101.94 ± 0.47 mg ml(-1) at 12 h PI in the 10(7) and 10(8) CFU ml(-1) groups, respectively. These data showed that both Hc content and TPCH varied as the same extent after infection. The up-regulation of the Hc content at 6-36 h PI might be a reference threshold for shrimp infection.

  11. Characterisation of the Bacterial and Fungal Communities Associated with Different Lesion Sizes of Dark Spot Syndrome Occurring in the Coral Stephanocoenia intersepta

    PubMed Central

    Sweet, Michael; Burn, Deborah; Croquer, Aldo; Leary, Peter

    2013-01-01

    The number and prevalence of coral diseases/syndromes are increasing worldwide. Dark Spot Syndrome (DSS) afflicts numerous coral species and is widespread throughout the Caribbean, yet there are no known causal agents. In this study we aimed to characterise the microbial communities (bacteria and fungi) associated with DSS lesions affecting the coral Stephanocoenia intersepta using nonculture molecular techniques. Bacterial diversity of healthy tissues (H), those in advance of the lesion interface (apparently healthy AH), and three sizes of disease lesions (small, medium, and large) varied significantly (ANOSIM R  = 0.052 p<0.001), apart from the medium and large lesions, which were similar in their community profile. Four bacteria fitted into the pattern expected from potential pathogens; namely absent from H, increasing in abundance within AH, and dominant in the lesions themselves. These included ribotypes related to Corynebacterium (KC190237), Acinetobacter (KC190251), Parvularculaceae (KC19027), and Oscillatoria (KC190271). Furthermore, two Vibrio species, a genus including many proposed coral pathogens, dominated the disease lesion and were absent from H and AH tissues, making them candidates as potential pathogens for DSS. In contrast, other members of bacteria from the same genus, such as V. harveyii were present throughout all sample types, supporting previous studies where potential coral pathogens exist in healthy tissues. Fungal diversity varied significantly as well, however the main difference between diseased and healthy tissues was the dominance of one ribotype, closely related to the plant pathogen, Rhytisma acerinum, a known causal agent of tar spot on tree leaves. As the corals’ symbiotic algae have been shown to turn to a darker pigmented state in DSS (giving rise to the syndromes name), the two most likely pathogens are R. acerinum and the bacterium Oscillatoria, which has been identified as the causal agent of the colouration in Black

  12. Coffin-Siris syndrome with café-au-lait spots, obesity and hyperinsulinism caused by a mutation in the ARID1B gene.

    PubMed

    Sonmez, Fatma Mujgan; Uctepe, Eyyup; Gunduz, Mehmet; Gormez, Zeliha; Erpolat, Seval; Oznur, Murat; Sagiroglu, Mahmut Samil; Demirci, Huseyin; Gunduz, Esra

    2016-08-01

    Coffin-Siris syndrome (CSS) (MIM 135900) is characterized by developmental delay, severe speech impairment, distinctive facial features, hypertrichosis, aplasia or hypoplasia of the distal phalanx or nail of the fifth digit and agenesis of the corpus callosum. Recently, it was shown that mutations in the ARID1B gene are the main cause of CSS, accounting for 76% of identified mutations. Here, we report a 15 year-old female patient who was admitted to our clinic with seizures, speech problems, dysmorphic features, bilaterally big, large thumb, café-au-lait (CAL) spots, obesity and hyperinsulinism. First, the patient was thought to have an association of neurofibromatosis and Rubinstein Taybi syndrome. Because of the large size of the NF1 gene for neurofibromatosis and CREBBP gene for Rubinstein Taybi syndrome, whole exome sequence analysis (WES) was conducted and a novel ARID1B mutation was identified. The proband WES test identified a novel heterozygous frameshift mutation c.3394_3395insTA in exon 13 of ARID1B (NM_017519.2) predicting a premature stop codon p.(Tyr1132Leufs*67). Sanger sequencing confirmed the heterozygous c.3394_3395insTA mutation in the proband and that it was not present in her parents indicating de novo mutation. Further investigation and new cases will help to understand this phenomenon better.

  13. Coffin-Siris syndrome with café-au-lait spots, obesity and hyperinsulinism caused by a mutation in the ARID1B gene

    PubMed Central

    Sonmez, Fatma Mujgan; Uctepe, Eyyup; Gunduz, Mehmet; Gormez, Zeliha; Erpolat, Seval; Oznur, Murat; Sagiroglu, Mahmut Samil; Demirci, Huseyin; Gunduz, Esra

    2016-01-01

    Summary Coffin-Siris syndrome (CSS) (MIM 135900) is characterized by developmental delay, severe speech impairment, distinctive facial features, hypertrichosis, aplasia or hypoplasia of the distal phalanx or nail of the fifth digit and agenesis of the corpus callosum. Recently, it was shown that mutations in the ARID1B gene are the main cause of CSS, accounting for 76% of identified mutations. Here, we report a 15 year-old female patient who was admitted to our clinic with seizures, speech problems, dysmorphic features, bilaterally big, large thumb, café-au-lait (CAL) spots, obesity and hyperinsulinism. First, the patient was thought to have an association of neurofibromatosis and Rubinstein Taybi syndrome. Because of the large size of the NF1 gene for neurofibromatosis and CREBBP gene for Rubinstein Taybi syndrome, whole exome sequence analysis (WES) was conducted and a novel ARID1B mutation was identified. The proband WES test identified a novel heterozygous frameshift mutation c.3394_3395insTA in exon 13 of ARID1B (NM_017519.2) predicting a premature stop codon p.(Tyr1132Leufs*67). Sanger sequencing confirmed the heterozygous c.3394_3395insTA mutation in the proband and that it was not present in her parents indicating de novo mutation. Further investigation and new cases will help to understand this phenomenon better. PMID:27672547

  14. Lymphoid organ cell culture system from Penaeus monodon (Fabricius) as a platform for white spot syndrome virus and shrimp immune-related gene expression.

    PubMed

    Jose, S; Jayesh, P; Sudheer, N S; Poulose, G; Mohandas, A; Philip, R; Singh, I S Bright

    2012-05-01

    Shrimp cell lines are yet to be reported and this restricts the prospects of investigating the associated viral pathogens, especially white spot syndrome virus (WSSV). In this context, development of primary cell cultures from lymphoid organs was standardized. Poly-l-lysine-coated culture vessels enhanced growth of lymphoid cells, while the application of vertebrate growth factors did not, except insulin-like growth factor-1 (IGF-1). Susceptibility of the lymphoid cells to WSSV was confirmed by immunofluoresence assay using monoclonal antibody against the 28 kDa envelope protein of WSSV. Expression of viral and immune-related genes in WSSV-infected lymphoid cultures could be demonstrated by RT-PCR. This emphasizes the utility of lymphoid primary cell culture as a platform for research in virus-cell interaction, virus morphogenesis, up and downregulation of shrimp immune-related genes, and also for the discovery of novel drugs to combat WSSV in shrimp culture.

  15. Cloning of profilin (FcPFN) from the shrimp Fenneropenaeus chinensis, a highly expressed protein in white spot syndrome virus (WSSV)-infected shrimp.

    PubMed

    Kong, H J; Hong, G-E; Cho, H K; Nam, B-H; Kim, Y-O; Kim, W-J; Lee, S-J; Kim, K-K

    2009-01-01

    We isolated and characterized the profilin (FcPFN) cDNA from hemocytes of Fenneropenaeus chinensis, a unique shrimp species from the Yellow Sea. The FcPFN cDNA consists of 830 bp and encodes a polypeptide of 125 amino acids, having a predicted isoelectric point of 5.06. The deduced amino acid sequence of FcPFN shows 36% and 90% amino acid sequence identity to the profilin genes of Pacific white shrimp Litopenaeus vannamei and black tiger shrimp Penaeus monodon, respectively. The FcPFN mRNA was highly expressed in hemocytes and hepatopancreas and moderately in muscle of normal shrimp. The higher expression of FcPFN mRNA is observed in shrimp infected with the white spot syndrome virus (WSSV), which is a major concern in all shrimp-growing regions of the world. These results suggest a potential role for FcPFN in viral host defense mechanisms.

  16. Expression profile of key immune-related genes in Penaeus monodon juveniles after oral administration of recombinant envelope protein VP28 of white spot syndrome virus.

    PubMed

    Thomas, Ancy; Sudheer, Naduvilamuriparampu Saidumuhammed; Kiron, Viswanath; Bright Singh, Issac S; Narayanan, Rangarajan Badri

    2016-07-01

    White spot syndrome virus (WSSV) is the most catastrophic pathogen the shrimp industry has ever encountered. VP28, the abundant envelope protein of WSSV was expressed in bacteria, the purified protein administered orally to Penaeus monodon juveniles and its immune modulatory effects examined. The results indicated significant up-regulation of caspase, penaeidin, crustin, astakine, syntenin, PmRACK, Rab7, STAT and C-type lectin in animals orally administered with this antigen. This revealed the immune modulations in shrimps followed by oral administration of rVP28P which resulted in the reduced transcription of viral gene vp28 and delay in mortality after WSSV challenge. The study suggests the potential of rVP28P to elicit a non-specific immune stimulation in shrimps.

  17. Competition of infectious hypodermal and haematopoietic necrosis virus (IHHNV) with white spot syndrome virus (WSSV) for binding to shrimp cellular membrane.

    PubMed

    Yan, D C; Huang, J; Yang, B; Sun, H S; Wang, Y Y; Liu, X

    2016-10-01

    Infectious hypodermal and haematopoietic necrosis virus (IHHNV) and white spot syndrome virus (WSSV) are two widespread shrimp viruses. The interference of IHHNV on WSSV was the first reported case of viral interference that involved crustacean viruses and has been subsequently confirmed. However, the mechanisms underlying the induction of WSSV resistance through IHHNV infection are practically unknown. In this study, the interference mechanisms between IHHNV and WSSV were studied using a competitive ELISA. The binding of WSSV and IHHNV to cellular membrane of Litopenaeus vannamei was examined. The results suggested that there existed a mutual competition between IHHNV and WSSV for binding to receptors present on cellular membrane of L. vannamei and that the inhibitory effects of WSSV towards IHHNV were more distinct than those of IHHNV towards WSSV.

  18. Immune response of white shrimp, Litopenaeus vannamei, after a concurrent infection with white spot syndrome virus and infectious hypodermal and hematopoietic necrosis virus.

    PubMed

    Yeh, Shinn-Pyng; Chen, Ying-Nan; Hsieh, Shu-Ling; Cheng, Winton; Liu, Chun-Hung

    2009-04-01

    In the present study, we investigated immunological changes in viral-infected white shrimp, Litopenaeus vannamei. White shrimp were infected with white spot syndrome virus (WSSV) or co-infected with WSSV and infectious hypodermal and hematopoietic necrosis virus (IHHNV) as detected by polymerase chain reaction (PCR). The complete (100%) mortality rate of shrimp was caused by viral infection due to immune parameters being suppressed including decreases in phenoloxidase activity, total hemocyte counts, differential hemocyte counts, and the gene expressions of prophenoloxidase and peroxinectin. In addition, increases in lipopolysaccharide and beta-1,3-glucan-binding protein of hemocytes and the hepatopancreas, and respiratory bursts per cell, and a decrease in superoxide dismutase were found in viral-infected shrimp, which may have been related to the defense against viral infection.

  19. Protection against white spot syndrome virus (WSSV) infection in kuruma shrimp orally vaccinated with WSSV rVP26 and rVP28.

    PubMed

    Satoh, Jun; Nishizawa, Toyohiko; Yoshimizu, Mamoru

    2008-11-20

    White spot syndrome virus (WSSV) is the causative agent of white spot disease (WSD), one of the most serious diseases affecting global shrimp farming. We compared WSSV infection induction in kuruma shrimp Marsupenaeus japonicus by oral, immersion, and intramuscular injection (IM) exposure methods and evaluated the oral vaccine prepared from the recombinant WSSV proteins rVP26 and rVP28. The 50% lethal doses (LD50) of WSSV by oral, immersion, and IM challenges were 10(-0.4), 109-4.4), and 10(-7.7) g shrimp(-1), respectively, indicating that WSSV infection efficiency by oral challenge was significantly less than the other 2 challenge routes. However, in shrimp farms it is believed that WSSV infection is easily and commonly established by the oral route as a result of cannibalization of WSSV-infected shrimp. Kuruma shrimp vaccinated orally with WSSV rVP26 or rVP28 were challenged with WSSV by oral, immersion, and IM routes to compare protection efficacy. The relative percent survival values were 100% for oral challenge, 70 to 71% for immersion, and 34 to 61% for IM. Thus, the protection against WSSV-infection that was induced in kuruma shrimp by oral vaccination with rVP26 or rVP28 seemed equivalent to that obtained through IM vaccination.

  20. Identification of RAPD-SCAR marker linked to white spot syndrome virus resistance in populations of giant black tiger shrimp, Penaeus monodon Fabricius.

    PubMed

    Dutta, S; Biswas, S; Mukherjee, K; Chakrabarty, U; Mallik, A; Mandal, N

    2014-05-01

    White spot disease (WSD) caused by white spot syndrome virus (WSSV) creates severe epizootics in shrimp aquaculture industry worldwide. Despite several efforts, no such permanent remedy was yet developed. Selective breeding using DNA markers would be a cost-effective strategy for long-term solution of this problem. In the present investigation, out of 30 random primers, only one primer produced a statistically significant (P < 0.01) randomly amplified polymorphic DNA (RAPD) marker of 502 bp, which provided a good discrimination between disease resistant and disease susceptible populations of Penaeus monodon from three geographical locations along the East coast of India. Because RAPD markers are dominant, a sequence characterized amplified region (SCAR) marker was developed by cloning and sequencing of 502 bp RAPD fragment, which generates a single 457 bp DNA fragment after PCR amplification only in the disease resistant shrimps. Challenge experiment was also conducted to validate this 457 bp SCAR marker, and the results suggested that the WSSV loads were 2.25 × 10(3) fold higher in disease susceptible than that in disease resistant shrimps using real-time PCR. Therefore, this 457 bp DNA SCAR marker will be very valuable towards the development of disease-free shrimp aquaculture industry.

  1. Development of a monoclonal antibody-based flow-through immunoassay (FTA) for detection of white spot syndrome virus (WSSV) in black tiger shrimp Penaeus monodon.

    PubMed

    Patil, R; Shankar, K M; Kumar, B T N; Kulkarni, A; Patil, P; Moger, N

    2013-09-01

    A flow-through immunoassay (FTA), an improved version of immunodot, was developed using a nitrocellulose membrane baked onto adsorbent pads enclosed in a plastic cassette to detect white spot syndrome virus (WSSV) in shrimp. Sharp purple dots developed with WSSV against the white background of the nitrocellulose membrane. The detection limits of WSSV by the FTA and immunodot were 0.312 and 1.2 μg mL(-1) crude WSSV protein, respectively. The FTA could be completed in 8-10 min compared with 90 min for immunodot. The FTA was 100 times more sensitive than 1-step polymerase chain reaction (PCR) and in between that of the 1- and 2-step PCR protocol recommended by the Office of International Epizootics (OIE). In experimental, orally infected shrimp post-larvae, WSSV was first detected 14, 16 and 18 h post-infection (hpi) by FTA, immunodot and one-step PCR, respectively. The FTA detected WSSV 2 and 4 h earlier than immunodot and one-step PCR, respectively. The FTA was more sensitive (25/27) than one-step PCR (23/27) and immunodot (23/27) for the detection of WSSV from white spot disease outbreak ponds. The reagent components of the FTA were stable giving expected results for 6 m at 4-8 °C. The FTA is available as a rapid test kit called 'RapiDot' for the early detection of WSSV under field conditions.

  2. Role of Marsupenaeus japonicus crustin-like peptide against Vibrio penaeicida and white spot syndrome virus infection.

    PubMed

    Hipolito, Sheryll Grospe; Shitara, Aiko; Kondo, Hidehiro; Hirono, Ikuo

    2014-10-01

    Crustins are important AMP that has been identified in crustaceans. In this study, the role of Marsupenaeus japonicus crustin-like peptide (MjCRS) was examined in vivo by RNA interference (RNAi) using double-stranded RNA (dsRNA). Tissue expression analysis revealed that MjCRS transcripts are expressed in different tissues tested with the highest expression observed in hemocytes. Treatment with double-stranded RNA specific to MjCRS led to a significant reduction of MjCRS transcripts within the hemocytes. When MjCRS was silenced and subsequently infected with Vibrio penaeicida final mortality was significantly higher compared with PBS and dsGFP treated groups. On the other hand, final mortalities of MjCRS silenced and PBS injected groups were not significantly different after infection with white spot virus, however, both are significantly higher compared with dsGFP treated group. V. penaeicida infection significantly decreased MjCRS expression at 3, 6, 12 and 24h followed by significant increase at 48 h post-infection. On the contrary, white spot infection significantly increased MjCRS expression at 6 and 12h and decreased at 48 h post-infection. dsRNA treatment alone decreased total hemocyte counts (THCs) and subsequent V. penaeicida or white spot virus infection further decreased THCs. VP28 gene expression was both similarly increased in PBS injected group and MjCRS silenced group at 24 and 48 h-post infection. Results suggest that MjCRS is involved in antibacterial defense and might not have critical function against viral infection.

  3. Susceptibility of juvenile European lobster Homarus gammarus to shrimp products infected with high and low doses of white spot syndrome virus.

    PubMed

    Bateman, K S; Munro, J; Uglow, B; Small, H J; Stentiford, G D

    2012-08-27

    White spot syndrome virus (WSSV) is the most important pathogen known to affect the sustainability and growth of the global penaeid shrimp farming industry. Although most commonly associated with penaeid shrimp farmed in warm waters, WSSV is also able to infect, cause disease in and kill a wide range of other decapod crustaceans, including lobsters, from temperate regions. In 2005, the European Union imported US$500 million worth of raw frozen or cooked frozen commodity products, much of which originated in regions positive for white spot disease (WSD). The presence of WSSV within the UK food market was verified by means of nested PCR performed on samples collected from a small-scale survey of supermarket commodity shrimp. Passage trials using inoculum derived from commodity shrimp from supermarkets and delivered by injection to specific pathogen-free Pacific white shrimp Litopenaeus vannamei led to rapid mortality and pathognomonic signs of WSD in the shrimp, demonstrating that WSSV present within commodity shrimp was viable. We exposed a representative European decapod crustacean, the European lobster Homarus gammarus, to a single feeding of WSSV-positive, supermarket-derived commodity shrimp, and to positive control material (L. vannamei infected with a high dose of WSSV). These trials demonstrated that lobsters fed positive control (high dose) frozen raw products succumbed to WSD and displayed pathognomonic signs associated with the disease as determined by means of histology and transmission electron microscopy. Lobsters fed WSSV-positive, supermarket-derived commodity shrimp (low dose) did not succumb to WSD (no mortality or pathognomonic signs of WSD) but demonstrated a low level or latent infection via PCR. This study confirms susceptibility of H. gammarus to WSSV via single feedings of previously frozen raw shrimp products obtained directly from supermarkets.

  4. Lateral flow assay for rapid detection of white spot syndrome virus (WSSV) using a phage-displayed peptide as bio-recognition probe.

    PubMed

    Kulabhusan, Prabir Kumar; Rajwade, Jyutika M; Sahul Hameed, A S; Paknikar, Kishore M

    2017-03-27

    White spot disease caused by the white spot syndrome virus (WSSV) has a major socio-economic impact on shrimp farming in India. It has been realized that a field-usable diagnostic capable of rapid detection of WSSV can prevent huge economic losses in disease outbreaks. In this work, we explored the possibility of using a peptide as bio-recognition probe in a field-usable device for the detection of WSSV from infected shrimps and prawns. A commercially available random phage-display library was screened against rVP28 (a major structural protein of WSSV, expressed as a recombinant protein in Escherichia coli). A bacteriophage clone VP28-4L was obtained, and its binding to purified rVP28 protein as well as WSSV from infected shrimp Litopaeneus vannamei tissue was confirmed by ELISA and western blot. The apparent equilibrium dissociation constant (Kd,app) was calculated to be 810 nM. VP28-4L did not show cross-reactivity with any other shrimp viruses. A 12-mer peptide (pep28, with the sequence 'TFQAFDLSPFPS') displayed on the VP28-4L was synthesized, and its diagnostic potential was evaluated in a lateral flow assay (LFA). Visual detection of WSSV could be achieved using biotinylated-pep28 and streptavidin-conjugated gold nanoparticles. In LFA, 12.5 μg/mL of the virus could be detected from L. vannamei gill tissue homogenate within 20 min. Pep28 thus becomes an attractive candidate in bio-recognition of WSSV in field-usable diagnostic platforms benefitting the aquaculture sector.

  5. Kinetic analysis of internalization of white spot syndrome virus by haemocyte subpopulations of penaeid shrimp, Litopenaeus vannamei (Boone), and the outcome for virus and cell.

    PubMed

    Tuan, V V; De Gryse, G M A; Thuong, K V; Bossier, P; Nauwynck, H J

    2016-12-01

    Little is known about the innate antiviral defence of shrimp haemocytes. In this context, the haemocytes of penaeid shrimp Litopenaeus vannamei (Boone) were separated by iodixanol density gradient centrifugation into five subpopulations (sub): sub 1 (hyalinocytes), sub 2 and 3 (prohyalinocytes), sub 4 (semigranulocytes) and sub 5 (granulocytes) and exposed to beads, white spot syndrome virus (WSSV) and ultraviolet (UV)-killed WSSV. In a first experiment, the uptake of beads, white spot syndrome virus (WSSV) and UV-killed WSSV by these different haemocyte subpopulations was investigated using confocal microscopy. Only haemocytes of sub 1, 4 and 5 were internalizing beads, WSSV and UV-killed WSSV. Beads were engulfed by a much larger percentage of cells (91.2 in sub 1; 84.1 in sub 4 and 58.1 in sub 5) compared to WSSV (9.6 in sub 1; 10.5 in sub 4 and 7.9 in sub 5) and UV-killed WSSV (12.9 in sub 1; 13.3 in sub 4; and 11.8 in sub 5). In a second experiment, it was shown that upon internalization, WSS virions lost their envelope most probably by fusion with the cellular membrane of the endosome (starting between 30 and 60 min post-inoculation) and that afterwards the capsid started to become disintegrated (from 360 min post-inoculation). Expression of new viral proteins was not observed. Incubation of haemocyte subpopulations with WSSV but not with UV-killed WSSV and polystyrene beads resulted in a significant drop in haemocyte viability. To find the underlying mechanism, a third experiment was performed in which haemocyte subpopulations were exposed to a short WSSV DNA fragment (VP19) and CpG ODNs. These small DNA fragments induced cell death. In conclusion, WSSV is efficiently internalized by hyalinocytes, semigranulocytes and granulocytes, after which the virus loses its envelope; as soon as the capsids start to disintegrate, cell death is activated, which in part may be explained by the exposure of viral DNA to cellular-sensing molecules.

  6. Enhanced survival of shrimp, Penaeus (Marsupenaeus) japonicus from white spot syndrome disease after oral administration of recombinant VP28 expressed in Brevibacillus brevis.

    PubMed

    Caipang, Christopher Marlowe A; Verjan, Noel; Ooi, Ei Lin; Kondo, Hidehiro; Hirono, Ikuo; Aoki, Takashi; Kiyono, Hiroshi; Yuki, Yoshikazu

    2008-09-01

    White spot syndrome virus (WSSV) disease is a major threat to shrimp culture worldwide. Here, we assessed the efficacy of the oral administration of purified recombinant VP28, an envelope protein of WSSV, expressed in a Gram-positive bacterium, Brevibacillus brevis, in providing protection in shrimp, Penaeus japonicus, upon challenge with WSSV. Juvenile shrimp (2-3g in body weight) fed with pellets containing purified recombinant VP28 (50microg/shrimp) for 2 weeks showed significantly higher survival rates than control groups when challenged with the virus at 3 days after the last day of feeding. However, when shrimp were challenged 2 weeks after the last day of feeding, survival rates decreased (33.4% and 24.93%, respectively). Survival rate was dose-dependent, increasing from 60.7 to 80.3% as the dose increased from 1 to 50microg/shrimp. At a dose of 50microg/shrimp, the recombinant protein provided protection as soon as 1 day after feeding (72.5% survival). Similar results were obtained with larger-sized shrimp. These results show that recombinant VP28 expressed in a Gram-positive bacterium is a potential oral vaccine against WSSV.

  7. Indel-II region deletion sizes in the white spot syndrome virus genome correlate with shrimp disease outbreaks in southern Vietnam.

    PubMed

    Hoa, Tran Thi Tuyet; Zwart, Mark P; Phuong, Nguyen T; Oanh, Dang T H; de Jong, Mart C M; Vlak, Just M

    2012-06-13

    Sequence comparisons of the genomes of white spot syndrome virus (WSSV) strains have identified regions containing variable-length insertions/deletions (i.e. indels). Indel-I and Indel-II, positioned between open reading frames (ORFs) 14/15 and 23/24, respectively, are the largest and the most variable. Here we examined the nature of these 2 indel regions in 313 WSSV-infected Penaeus monodon shrimp collected between 2006 and 2009 from 76 aquaculture ponds in the Mekong Delta region of Vietnam. In the Indel-I region, 2 WSSV genotypes with deletions of either 5950 or 6031 bp in length compared with that of a reference strain from Thailand (WSSV-TH-96-II) were detected. In the Indel-II region, 4 WSSV genotypes with deletions of 8539, 10970, 11049 or 11866 bp in length compared with that of a reference strain from Taiwan (WSSV-TW) were detected, and the 8539 and 10970 bp genotypes predominated. Indel-II variants with longer deletions were found to correlate statistically with WSSV-diseased shrimp originating from more intensive farming systems. Like Indel-I lengths, Indel-II lengths also varied based on the Mekong Delta province from which farmed shrimp were collected.

  8. Two new anti-apoptotic proteins of white spot syndrome virus that bind to an effector caspase (PmCasp) of the giant tiger shrimp Penaeus (Penaeus) monodon.

    PubMed

    Lertwimol, Tareerat; Sangsuriya, Pakkakul; Phiwsaiya, Kornsunee; Senapin, Saengchan; Phongdara, Amornrat; Boonchird, Chuenchit; Flegel, Timothy W

    2014-05-01

    White spot syndrome virus proteins WSSV134 and WSSV322 have been shown to bind with the p20 domain (residues 55-214) of Penaeus monodon caspase (PmCasp) protein through yeast two-hybrid screening. Binding was confirmed for the p20 domain and the full-length caspase by co-immunoprecipitation. WSSV134 is also known as the WSSV structural protein VP36A, but no function or conserved domains have been ascribed to WSSV322. Discovery of the caspase binding activity of these two proteins led to an investigation of their possible anti-apoptotic roles. Full-length PmCasp was confirmed to be an effector caspase by inducing apoptosis in transfected Sf-9 cells as assessed by DAPI staining. Using the same cell model, comparison of cells co-transfected with PmCasp and either WSSV134 or WSSV322 revealed that both of the binding proteins had anti-apoptotic activity. However, using the same Sf-9 protocol with anti-apoptosis protein-1 (AAP-1; also called WSSV449) previously shown to bind and inactivate a different effector caspase from P. monodon (Pm caspase) did not block apoptosis induced by PmCasp. The results revealed diversity in effector caspases and their viral protein inhibitors in P. monodon.

  9. Protection of shrimp against white spot syndrome virus (WSSV) with β-1,3-D-glucan-encapsulated vp28-siRNA particles.

    PubMed

    Zhu, Fei; Zhang, Xiaobo

    2012-02-01

    White spot syndrome virus (WSSV) is a major shrimp viral pathogen responsible for large economic losses to shrimp aquaculture all over the world. The RNAi mediated by siRNA contributes a new strategy to control this viral disease. However, the efficient approach to deliver the siRNA into shrimp remains to be addressed. In this investigation, an antiviral vp28-siRNA was encapsulated in β-1,3-D-glucan, and then the β-1,3-D-glucan-encapsulated vp28-siRNA particles (GeRPs) were delivered into Marsupenaeus japonicus shrimp. The results showed that the vp28-siRNA in GeRPs could be released in hemocytes of shrimp. It was found that the GeRPs containing the vp28-siRNA inhibited the replication of WSSV in vivo, which presented a better antiviral activity than the non-encapsulated vp28-siRNA. Further evidence indicated that the mortality of WSSV-infected shrimp was significantly delayed by the GeRPs containing vp28-siRNA. Therefore, our study presented that the glucan-encapsulated siRNA might represent a novel potential therapeutic or preventive approach to control the shrimp disease.

  10. PmVRP15, a novel viral responsive protein from the black tiger shrimp, Penaeus monodon, promoted white spot syndrome virus replication.

    PubMed

    Vatanavicharn, Tipachai; Prapavorarat, Adisak; Jaree, Phattarunda; Somboonwiwat, Kunlaya; Tassanakajon, Anchalee

    2014-01-01

    Suppression subtractive hybridization of Penaeus monodon hemocytes challenged with white spot syndrome virus (WSSV) has identified the viral responsive gene, PmVRP15, as the highest up-regulated gene ever reported in shrimps. Expression analysis by quantitative real time RT-PCR revealed 9410-fold up-regulated level at 48 h post WSSV injection. Tissue distribution analysis showed that PmVRP15 transcript was mainly expressed in the hemocytes of shrimp. The full-length cDNA of PmVRP15 transcript was obtained and showed no significant similarity to any known gene in the GenBank database. The predicted open reading frame of PmVRP15 encodes for a deduced 137 amino acid protein containing a putative transmembrane helix. Immunofluorescent localization of the PmVRP15 protein revealed it accumulated around the nuclear membrane in all three types of shrimp hemocytes and that the protein was highly up-regulated in WSSV-infected shrimps. Double-stranded RNA interference-mediated gene silencing of PmVRP15 in P. monodon significantly decreased WSSV propagation compared to the control shrimps (injected with GFP dsRNA). The significant decrease in cumulative mortality rate of WSSV-infected shrimp following PmVRP15 knockdown was observed. These results suggest that PmVRP15 is likely to be a nuclear membrane protein and that it acts as a part of WSSV propagation pathway.

  11. Co-interactive DNA-binding between a novel, immunophilin-like shrimp protein and VP15 nucleocapsid protein of white spot syndrome virus.

    PubMed

    Sangsuriya, Pakkakul; Senapin, Saengchan; Huang, Wei-Pang; Lo, Chu-Fang; Flegel, Timothy W

    2011-01-01

    White spot syndrome virus (WSSV) is one of the most serious pathogens of penaeid shrimp. Although its genome has been completely characterized, the functions of most of its putative proteins are not yet known. It has been suggested that the major nucleocapsid protein VP15 is involved in packaging of the WSSV genome during virion formation. However, little is known in its relationship with shrimp host cells. Using the yeast two-hybrid approach to screen a shrimp lymphoid organ (LO) cDNA library for proteins that might interact with VP15, a protein named PmFKBP46 was identified. It had high sequence similarity to a 46 kDa-immunophilin called FKBP46 from the lepidopteran Spodoptera frugiperda (the fall armyworm). The full length PmFKBP46 consisted of a 1,257-nucleotide open reading frame with a deduced amino acid sequence of 418 residues containing a putative FKBP-PPIase domain in the C-terminal region. Results from a GST pull-down assay and histological co-localization revealed that VP15 physically interacted with PmFKBP46 and that both proteins shared the same subcellular location in the nucleus. An electrophoretic mobility shift assay indicated that PmFKBP46 possessed DNA-binding activity and functionally co-interacted with VP15 in DNA binding. The overall results suggested that host PmFKBP46 might be involved in genome packaging by viral VP15 during virion assembly.

  12. Sulfated galactans isolated from the red seaweed Gracilaria fisheri target the envelope proteins of white spot syndrome virus and protect against viral infection in shrimp haemocytes.

    PubMed

    Rudtanatip, Tawut; Asuvapongpatana, Somluk; Withyachumnarnkul, Boonsirm; Wongprasert, Kanokpan

    2014-05-01

    The present study was aimed at evaluating an underlying mechanism of the antiviral activity of the sulfated galactans (SG) isolated from the red seaweed Gracilaria fisheri against white spot syndrome virus (WSSV) infection in haemocytes of the black tiger shrimp Penaeus monodon. Primary culture of haemocytes from Penaeus monodon was performed and inoculated with WSSV, after which the cytopathic effect (CPE), cell viability and viral load were determined. Haemocytes treated with WSSV-SG pre-mix showed decreased CPE, viral load and cell mortality from the viral infection. Solid-phase virus-binding assays revealed that SG bound to WSSV in a dose-related manner. Far Western blotting analysis indicated that SG bound to VP 26 and VP 28 proteins of WSSV. In contrast to the native SG, desulfated SG did not reduce CPE and cell mortality, and showed low binding activity with WSSV. The current study suggests that SG from Gracilaria fisheri elicits its anti-WSSV activity by binding to viral proteins that are important for the process of viral attachment to the host cells. It is anticipated that the sulfate groups of SG are important for viral binding.

  13. White spot syndrome virus entry is dependent on multiple endocytic routes and strongly facilitated by Cq-GABARAP in a CME-dependent manner

    PubMed Central

    Chen, Rong-yuan; Shen, Kai-li; Chen, Zhen; Fan, Wei-wei; Xie, Xiao-lu; Meng, Chuang; Chang, Xue-jiao; Zheng, Li-bing; Jeswin, Joseph; Li, Cheng-hua; Wang, Ke-jian; Liu, Hai-peng

    2016-01-01

    White spot syndrome virus (WSSV) is a lethal pathogen of shrimp and many other crustaceans, including crayfish. However, the molecular mechanism underlying its cellular entry remains elusive due to the lack of shrimp cell lines for viral propagation. Crayfish hematopoietic tissue (Hpt) cell culture was recently established as a good model for WSSV infection study. Here, we showed that multiple endocytic routes, including clathrin-mediated endocytosis (CME), macropinocytosis and caveolae-mediated endocytosis, were indispensably employed for the viral entry into Hpt cell of the crayfish Cherax quadricarinatus. Intriguingly, cellular autophagic activity was positively correlated with efficient viral entry, in which a key autophagy-related protein, γ-aminobutyric acid receptor-associated protein (Cq-GABARAP), that not only localized but also co-localized with WSSV on the Hpt cell membrane, strongly facilitated WSSV entry by binding to the viral envelope VP28 in a CME-dependent manner that was negatively regulated by Cq-Rac1. Furthermore, cytoskeletal components, including Cq-β-tubulin and Cq-β-actin, bound to both recombinant rCq-GABARAP and WSSV envelope proteins, which likely led to viral entry promotion via cooperation with rCq-GABARAP. Even under conditions that promoted viral entry, rCq-GABARAP significantly reduced viral replication at an early stage of infection, which was probably caused by the formation of WSSV aggregates in the cytoplasm. PMID:27385304

  14. A routine method for cholesterol and 7-dehydrocholesterol analysis in dried blood spot by GC-FID to diagnose the Smith-Lemli-Opitz syndrome.

    PubMed

    Gelzo, Monica; Clericuzio, Stefano; Barone, Rosalba; D'Apolito, Oceania; Dello Russo, Antonio; Corso, Gaetano

    2012-10-15

    This work was aimed to implement a fast and simple method to quantify cholesterol (CHOL) and 7-dehydrocholesterol (7-DHC) in dried blood spot (DBS) to diagnose the Smith-Lemli-Opitz syndrome (SLOS), an inborn error of CHOL biosynthesis. We developed and validated a GC-FID method for separation and quantification of underivatized CHOL and 7-DHC using a DBS disc of 6mm with a run time of 9 min. Correlation coefficients (r) of calibration curves ranged from 0.998 to 0.999 for CHOL and from 0.997 to 0.998 for 7-DHC. Within-day and between-day imprecision (CV%), accuracy (%), carry-over, and extraction efficacy (%) were also evaluated for validation. CHOL and 7-DHC were analyzed in DBS and plasma samples from 8 SLOS patients and 30 unaffected subjects. In SLOS patients, 7-DHC/CHOL ratios in DBS and plasma samples ranged from 0.035 to 1.448 and from 0.012 to 0.926, respectively. Results from calibration curves, quality controls and patient samples reveal that the method is suitable to analyze DBS to screen patients affected by SLOS.

  15. Identification of a Novel Nonstructural Protein, VP9, from White Spot Syndrome Virus: Its Structure Reveals a Ferredoxin Fold with Specific Metal Binding Sites

    SciTech Connect

    Liu,Y.; Wu, J.; Song, J.; Sivaraman, J.; Hew, C.

    2006-01-01

    White spot syndrome virus (WSSV) is a major pathogen in shrimp aquaculture. VP9, a full-length protein of WSSV, encoded by open reading frame wsv230, was identified for the first time in the infected Penaeus monodon shrimp tissues, gill, and stomach as a novel, nonstructural protein by Western blotting, mass spectrometry, and immunoelectron microscopy. Real-time reverse transcription-PCR demonstrated that the transcription of VP9 started from the early to the late stage of WSSV infection as a major mRNA species. The structure of full-length VP9 was determined by both X-ray and nuclear magnetic resonance (NMR) techniques. It is the first structure to be reported for WSSV proteins. The crystal structure of VP9 revealed a ferredoxin fold with divalent metal ion binding sites. Cadmium sulfate was found to be essential for crystallization. The Cd2+ ions were bound between the monomer interfaces of the homodimer. Various divalent metal ions have been titrated against VP9, and their interactions were analyzed using NMR spectroscopy. The titration data indicated that VP9 binds with both Zn2+ and Cd2+. VP9 adopts a similar fold as the DNA binding domain of the papillomavirus E2 protein. Based on our present investigations, we hypothesize that VP9 might be involved in the transcriptional regulation of WSSV, a function similar to that of the E2 protein during papillomavirus infection of the host cells.

  16. Generation of recombinant monoclonal antibodies to study structure-function of envelope protein VP28 of white spot syndrome virus from shrimp

    SciTech Connect

    Wang Yuzhen; Zhang Xiaohua; Yuan Li; Xu Tao; Rao Yu; Li Jia; Dai Heping

    2008-08-08

    White spot syndrome virus (WSSV) is a major pathogen in shrimp aquaculture. VP28 is one of the most important envelope proteins of WSSV. In this study, a recombinant antibody library, as single-chain fragment variable (scFv) format, displayed on phage was constructed using mRNA from spleen cells of mice immunized with full-length VP28 expressed in Escherichia coli. After several rounds of panning, six scFv antibodies specifically binding to the epitopes in the N-terminal, middle, and C-terminal regions of VP28, respectively, were isolated from the library. Using these scFv antibodies as tools, the epitopes in VP28 were located on the envelope of the virion by immuno-electron microscopy. Neutralization assay with these antibodies in vitro suggested that these epitopes may not be the attachment site of WSSV to host cell receptor. This study provides a new way to investigate the structure and function of the envelope proteins of WSSV.

  17. Analysis of the VP19 and VP28 genes of white spot syndrome virus in Korea and comparison with strains from other countries.

    PubMed

    Park, Jung-Eun; Shin, Hyun-Jin

    2009-01-01

    The results of our DNA analysis showed that there was 100% homology of the VP28 and VP19 genes of white spot syndrome virus (WSSV) among the samples collected from different shrimp farms in Korea. Comparing with an earlier isolated Korean strain, Korea01, the genes nucleotide sequences had only a single base difference which was observed in both VP19 and VP28. This resulted in a single amino acid substitution at position 40 of the latter. This implies that a single genetic strain of WSSV has been circulating in Korea and that its mutation rate is very low. In comparison with known sequences of VP19 and VP28 genes of WSSV isolates from other countries, the Korean strains had more than 99% sequence homology with those in gene and protein. Based on our sequence analysis of VP19 and VP28 of WSSV from various shrimp farms in Korea, the WSSV strains circulating in the region were genetically identical and similar to the strain identified two years ago. In addition, the Korean strain had close genetic identity with strains circulating in other Asian countries as well as other continents.

  18. Molecular docking and simulation studies of 3-(1-chloropiperidin-4-yl)-6-fluoro benzisoxazole 2 against VP26 and VP28 proteins of white spot syndrome virus.

    PubMed

    Sudharsana, S; Rajashekar Reddy, C B; Dinesh, S; Rajasekhara Reddy, S; Mohanapriya, A; Itami, T; Sudhakaran, R

    2016-10-01

    White spot syndrome virus (WSSV), an aquatic virus infecting shrimps and other crustaceans, is widely distributed in Asian subcontinents including India. The infection has led to a serious economic loss in shrimp farming. The WSSV genome is approximately 300 kb and codes for several proteins mediating the infection. The envelope proteins VP26 and VP28 play a major role in infection process and also in the interaction with the host cells. A comprehensive study on the viral proteins leading to the development of safe and potent antiviral therapeutic is of adverse need. The novel synthesized compound 3-(1-chloropiperidin-4-yl)-6-fluoro benzisoxazole 2 is proved to have potent antiviral activity against WSSV. The compound antiviral activity is validated in freshwater crabs (Paratelphusa hydrodomous). An in silico molecular docking and simulation analysis of the envelope proteins VP26 and VP28 with the ligand 3-(1-chloropiperidin-4-yl)-6-fluoro benzisoxazole 2 are carried out. The docking analysis reveals that the polar amino acids in the pore region of the envelope proteins were involved in the ligand binding. The influence of the ligand binding on the proteins is validated by the molecular dynamics and simulation study. These in silico approaches together demonstrate the ligand's efficiency in preventing the trimers from exhibiting their physiological function.

  19. Proteomic analysis by iTRAQ in red claw crayfish, Cherax quadricarinatus, hematopoietic tissue cells post white spot syndrome virus infection.

    PubMed

    Jeswin, Joseph; Xie, Xiao-lu; Ji, Qiao-lin; Wang, Ke-jian; Liu, Hai-peng

    2016-03-01

    To elucidate proteomic changes of Hpt cells from red claw crayfish, Cherax quadricarinatus, we have carried out isobaric tags for relative and absolute quantitation (iTRAQ) of cellular proteins at both early (1 hpi) and late stage (12 hpi) post white spot syndrome virus (WSSV) infection. Protein database search revealed 594 protein hits by Mascot, in which 17 and 30 proteins were present as differentially expressed proteins at early and late viral infection, respectively. Generally, these differentially expressed proteins include: 1) the metabolic process related proteins in glycolysis and glucogenesis, DNA replication, nucleotide/amino acid/fatty acid metabolism and protein biosynthesis; 2) the signal transduction related proteins like small GTPases, G-protein-alpha stimulatory subunit, proteins bearing PDZ- or 14-3-3-domains that help holding together and organize signaling complexes, casein kinase I and proteins of the MAP-kinase signal transduction pathway; 3) the immune defense related proteins such as α-2 macroglobulin, transglutaminase and trans-activation response RNA-binding protein 1. Taken together, these protein information shed new light on the host cellular response against WSSV infection in a crustacean cell culture.

  20. Effect of multiple infections with white spot syndrome virus and Vibrio anguillarum on Pacific white shrimp Litopenaeus vannamei (L.): mortality and viral replication.

    PubMed

    Jang, I K; Qiao, G; Kim, S-K

    2014-10-01

    Multiple infections are commonly found in practical shrimp culture and may cause more serious consequences than infections by one pathogen only. Therefore, this study was conducted to evaluate the effect of multiple infections with white spot syndrome virus (WSSV) and Vibrio anguillarum on Pacific white shrimp Litopenaeus vannamei (L.), mortality, WSSV replication in vivo and host immune response. In the WSSV single-infection group (WSSV load, 2 × 10(2) copies μL(-1)), mean cumulative mortality was 29.2%. In the V. anguillarum single-infection group, cumulative mortality was 12.5% when shrimp were challenged by 10(5) CFU mL(-1) of bacteria. In the co- and super-infection groups, 37.5% and 50% cumulative mortalities, respectively, were observed at a lower bacterial concentration of 10(3) CFU mL(-1), suggesting that shrimp with multiple infections died earlier and more frequently than singly infected shrimp. WSSV load after injection was tracked over time by TaqMan quantitative PCR. WSSV load increased more rapidly in the multiple-infection groups than in the single-infection group. Additionally, mRNA expression of the genes encoding prophenoloxidase 1 and 2, which are closely involved in innate immunity in shrimp, was down-regulated more extensively in multiple-infection groups than in single-infection groups, as indicated by quantitative reverse-transcription PCR.

  1. Regulation of the immediate-early genes of white spot syndrome virus by Litopenaeus vannamei kruppel-like factor (LvKLF).

    PubMed

    Huang, Ping-Han; Lu, Shao-Chia; Yang, Shu-Han; Cai, Pei-Si; Lo, Chu-Fang; Chang, Li-Kwan

    2014-10-01

    Kruppel-like factors (KLFs) belong to a subclass of Cys2/His2 zinc-finger DNA-binding proteins, and act as important regulators with diverse roles in cell growth, proliferation, differentiation, apoptosis and tumorigenesis. Our previous research showed that PmKLF from Penaeus monodon is crucial for white spot syndrome virus (WSSV) infection, yet the mechanisms by which PmKLF influences WSSV infection remain unclear. This study cloned KLF from Litopenaeus vannamei (LvKLF), which had 93% similarity with PmKLF. LvKLF formed a dimer via the C-terminal zinc-finger motif. Knockdown of LvKLF expression by dsRNA injection in WSSV-challenged shrimps was found to significantly inhibit the transcription of two important immediate-early (IE) genes, IE1 and WSSV304, and also reduced WSSV copy numbers. Moreover, reporter assays revealed that the promoter activities of these two WSSV IE genes were substantially enhanced by LvKLF. Mutations introduced in the promoter sequences of IE1 and WSSV304 were shown to abolish LvKLF activation of promoter activities; and an electrophoretic mobility shift assay demonstrated that LvKLF binds to putative KLF-response elements (KRE) in the promoters. Taken together, these results indicate that LvKLF transcriptional regulation of key IE genes is critical to WSSV replication.

  2. PmTBC1D20, a Rab GTPase-activating protein from the black tiger shrimp, Penaeus monodon, is involved in white spot syndrome virus infection.

    PubMed

    Yingvilasprasert, Wanchart; Supungul, Premruethai; Tassanakajon, Anchalee

    2014-02-01

    TBC (TRE2/BUB2/CDC16) domain proteins contain an ≈ 200-amino-acid motif and function as Rab GTPase-activating proteins that are required for regulating the activity of Rab proteins, and so, in turn, endocytic membrane trafficking in cells. TBC domain family member 20 (TBC1D20) has recently been reported to mediate Hepatitis C virus replication. Herein, PmTBC1D20 identified from the black tiger shrimp, Penaeus monodon, was characterized and evaluated for its role in white spot syndrome virus (WSSV) infection. The full-length cDNA sequence of PmTBC1D20 contains 2003 bp with a predicted 1443 bp open reading frame encoding a deduced 480 amino acid protein. Its transcript levels were significantly up-regulated at 24 and 48 h by ≈ 2.3- and 2.1-fold, respectively, after systemic infection with WSSV. In addition, depletion of PmTBC1D20 transcript in shrimps by double stranded RNA interference led to a decrease in the level of transcripts of three WSSV genes (VP28, ie1 and wsv477). This suggests the importance of PmTBC1D20 in WSSV infection. This is the first report of TBC1D20 in a crustacean and reveals the possible mechanism used by WSSV to modulate the activity of the host protein, PmTBC1D20, for its benefit in viral trafficking and replication.

  3. Molecular characterization of LvAV in response to white spot syndrome virus infection in the Pacific white shrimp (Litopenaeus vannamei).

    PubMed

    He, Shulin; Song, Lei; Qian, Zhaoying; Hou, Fujun; Liu, Yongjie; Wang, Xianzong; Peng, Zhangming; Sun, Chengbo; Liu, Xiaolin

    2015-07-01

    Litopenaeus vannamei is the most important farmed shrimp species globally, but its production is affected by several factors, including infectious disease. White spot syndrome virus (WSSV), in particular, causes significant shrimp losses. To understand the shrimp's immune response against WSSV, we cloned LvAV from L. vannamei and analyzed its expression pattern in different tissues, in addition to its expression following infection. We employed dsRNA and recombinant (r)LvAV to explore the potential role of LvAV in shrimp immunity when infected with WSSV. We find that LvAV is a C-type Lectin composed of 176 amino acids with a signal peptide and a specific C-type Lectin-type domain (CTLD). It shares 81% amino acid similarity with PmAV, an antiviral-like C-type Lectin from Penaeus monodom, and it is highly expressed in the hepatopancreas. Its expression is affected by infection with both WSSV and V. parahaemolyticus. Significantly, injection with rLvAV slowed WSSV replication, while injection with LvAV dsRNA initially led to enhanced virus propagation. Surprisingly, LvAV dsRNA subsequently led to a dramatic decrease in viral load in the later stages of infection, suggesting that LvAV may be subverted by WSSV to enhance viral replication or immune avoidance. Our results indicate that LvAV plays an important, but potentially complex role in the Pacific white shrimp's immune defense.

  4. Cloning and characterization of cytoplasmic dynein intermediate chain in Fenneropenaeus chinensis and its essential role in white spot syndrome virus infection.

    PubMed

    Feng, Jixing; Tang, Xiaoqian; Zhan, Wenbin

    2014-08-01

    To investigate the role of cytoplasmic dynein in white spot syndrome virus (WSSV) infection, the full-length cDNA of cytoplasmic dynein intermediate chain (FcDYNCI) was cloned in Fenneropenaeus chinensis, which consists of 2582 bp and encodes a polypeptide of 660 amino acids. Sequence analysis and multiple sequence alignment displayed that FcDYNCI was a member of cytoplasmic dynein 1 family. The FcDYNCI mRNA was most highly expressed in hemocytes, which was significantly up-regulated post WSSV infection. At 12 h post infection (hpi), confocal microscopic observation showed that WSSV could be co-localized with cytoplasmic dynein in hemocytes. After silencing by specific FcDYNCI dsRNA, the FcDYNCI mRNA level and the protein amount of FcDYNCI in hemocytes both exhibited a significant reduction, and the expression levels of three WSSV genes ie1, wsv477 and vp28 all exhibited the greatest decreases at 24 hpi. These results suggested that cytoplasmic dynein was involved in WSSV infection.

  5. The expression and purification of WSSV134 from white spot syndrome virus and its inhibitory effect on caspase activity from Penaeus monodon.

    PubMed

    Bowornsakulwong, Thanunthon; Charoensapsri, Walaiporn; Rattanarojpong, Triwit; Khunrae, Pongsak

    2017-02-01

    WSSV134 or VP36A protein of white spot syndrome virus was previously reported to be able to reduce apoptosis in Sf-9 cells transfected with caspase of Penaeus monodon (PmCasp). The protein was therefore believed to have a role in supporting the survival of WSSV inside the host cells during infection. However, the anti-apoptosis activity of WSSV134 involved in the inhibition of PmCasp is still unclear. In this study, we produced a recombinant WSSV134 (rWSSV134) and tested for its ability to inhibit PmCasp in vitro. The results from a caspase inhibition assay revealed that rWSSV134 could inhibit PmCasp in a dose-dependent manner. Since WSSV134 was predicted to contain three potential caspase binding sites, corresponding to the D54, D104 and D259, we then employed site-directed mutagenesis to investigate the involvement of these sites in PmCasp inhibition. D54A and D259A mutants could still inhibit PmCasp while D104A mutant lacks this activity. Our results confirmed that the WSSV134 is an inhibitor for PmCasp and that residue D104 is important for PmCasp inhibition.

  6. The promoter of the white spot syndrome virus immediate-early gene WSSV108 is activated by the cellular KLF transcription factor.

    PubMed

    Liu, Wang-Jing; Lo, Chu-Fang; Kou, Guang-Hsiung; Leu, Jiann-Horng; Lai, Ying-Jang; Chang, Li-Kwan; Chang, Yun-Shiang

    2015-03-01

    A series of deletion and mutation assays of the white spot syndrome virus (WSSV) immediate-early gene WSSV108 promoter showed that a Krüppel-like factor (KLF) binding site located from -504 to -495 (relative to the transcription start site) is important for the overall level of WSSV108 promoter activity. Electrophoretic mobility shift assays further showed that overexpressed recombinant Penaeus monodon KLF (rPmKLF) formed a specific protein-DNA complex with the (32)P-labeled KLF binding site of the WSSV108 promoter, and that higher levels of Litopenaeus vannamei KLF (LvKLF) were expressed in WSSV-infected shrimp. A transactivation assay indicated that the WSSV108 promoter was strongly activated by rPmKLF in a dose-dependent manner. Lastly, we found that specific silencing of LvKLF expression in vivo by dsRNA injection dramatically reduced both WSSV108 expression and WSSV replication. We conclude that shrimp KLF is important for WSSV genome replication and gene expression, and that it binds to the WSSV108 promoter to enhance the expression of this immediate-early gene.

  7. Expression profile of bio-defense genes in Penaeus monodon gills in response to formalin inactivated white spot syndrome virus vaccine.

    PubMed

    Sudheer, N S; Poulose, Gigi; Thomas, Ancy; Viswanath, Kiron; Kulkarni, Amod; Narayanan, R B; Philip, Rosamma; Singh, I S Bright

    2015-05-01

    White spot syndrome virus (WSSV) is the most devastating pathogen of penaeid shrimp. While developing technology to vaccinate shrimp against WSSV, it is imperative to look into the immune response of the animal at molecular level. However, very little information has been generated in this direction. The present study is an attempt to understand the expression of bio-defense genes in gill tissues of Penaeus monodon in response to formalin inactivated WSSV. A WSSV vaccine with a viral titer of 1×10(9) DNA copies was prepared and orally administered to P. monodon at a rate of 1.75×10(6) DNA copies of inactivated virus preparation (IVP) day(-1) for 7days. The animals were challenged with WSSV on 1st and 5th day post vaccination, and temporal expression of bio-defense genes in gill tissues was studied. Survival of 100% and 50% were observed respectively on 1st and 5th day post vaccination challenge. The humoral immune genes prophenoloxidase (proPO), alpha 2-macroglobulin (α2M), crustin and PmRACK, and the cell mediated immune genes caspase and Rab7 were up regulated in gill tissue upon vaccination and challenge. The expression of humoral gene crustin and cellular gene Rab7 was related to survival in IVP administered shrimp. Results of the study suggest that these genes have roles in protecting shrimp from WSSV on vaccination.

  8. Identification of highly expressed host microRNAs that respond to white spot syndrome virus infection in the Pacific white shrimp Litopenaeus vannamei (Penaeidae).

    PubMed

    Zeng, D G; Chen, X L; Xie, D X; Zhao, Y Z; Yang, Q; Wang, H; Li, Y M; Chen, X H

    2015-05-11

    MicroRNAs (miRNAs) are known to play an important role in regulating both adaptive and innate immunity. Pacific white shrimp (Litopenaeus vannamei) is the most widely farmed crustacean species in the world. However, little is known about the role miRNAs play in shrimp immunity. To understand the impact of viral infection on miRNA expression in shrimp, we used high-throughput sequencing technology to sequence two small RNA libraries prepared from L. vannamei under normal and white spot syndrome virus (WSSV) challenged conditions. Approximately 19,312,189 and 39,763,551 raw reads corresponding to 17,414,787 and 28,633,379 high-quality mappable reads were obtained from the two libraries, respectively. Twelve conserved miRNAs and one novel miRNA that were highly expressed (>100 RPM) in L. vannamei were identified. Of the identified miRNAs, 8 were differentially expressed in response to the virus infection, of which 1 was upregulated and 7 were downregulated. The prediction of miRNA targets showed that the target genes of the differentially expressed miRNAs were related to immunity, apoptosis, and development functions. Our study provides the first characterization of L. vannamei miRNAs in response to WSSV infection, which will help to reveal the roles of miRNAs in the antiviral mechanisms of shrimp.

  9. Shrimp STAT was hijacked by white spot syndrome virus immediate-early protein IE1 involved in modulation of viral genes.

    PubMed

    Yao, Defu; Ruan, Lingwei; Lu, Huasong; Shi, Hong; Xu, Xun

    2016-12-01

    STATs are a family of transcription factors that regulate a cascade of cellular processes including cell growth, differentiation, apoptosis and immune responses. However, they are usually targeted by viruses to assist infection. In this study, we identified that white spot syndrome virus (WSSV) immediate-early protein IE1 interacted with Litopenaeus vannamei STAT (LvSTAT) and thereby led to its phosphorylation activation. In addition, we demonstrated that LvSTAT could bind to the promoters of the viral immediate-early genes wsv051 and ie1 through STAT-binding motifs in vitro and vivo, allowing the enhancement of their promoters' activities. Moreover, IE1 could promote the transcriptional activation activity of LvSTAT to augment the transcription of wsv051 and ie1. In conclusion, our findings revealed a novel linkage between WSSV IE1 and shrimp STAT, which was a clue to well understand how WSSV adopted the active strategies to modulate the shrimp signaling pathway.

  10. Immunostimulatory activity of sulfated galactans isolated from the red seaweed Gracilaria fisheri and development of resistance against white spot syndrome virus (WSSV) in shrimp.

    PubMed

    Wongprasert, Kanokpan; Rudtanatip, Tawut; Praiboon, Jantana

    2014-01-01

    Sulfated galactans (SG) were isolated from the red seaweed Gracilaria fisheri (G. fisheri). Chemical analysis revealed SG contains sulfate (12.7%) and total carbohydrate (42.2%) with an estimated molecular mass of 100 kDa. Structure analysis by NMR and FT-IR spectroscopy revealed that SG is a complex structure with a linear backbone of alternating 3-linked β-D-galactopyranose and 4-linked 3,6-anhydrogalactose units with partial 6-O-methylate-β-D-galactopyranose and with sulfation occurring on C4 of D-galactopyranose and C6 of L-galactopyranose units. SG treatment enhanced immune parameters including total haemocytes, phenoloxidase activity, superoxide anions and superoxide dismutase in shrimp Penaeus monodon. Shrimp fed with Artemia salina enriched with SG (100 and 200 μg ml(-1)) and inoculated with white spot syndrome virus (WSSV) showed a significantly lower mortality rate and lower viral VP 28 amplification and expression than control. The results suggest that SG from G. fisheri exhibits immune stimulatory and antiviral activities that could protect P. monodon from WSSV infection.

  11. White spot syndrome virus IE1 and WSV056 modulate the G1/S transition by binding to the host retinoblastoma protein.

    PubMed

    Ran, Xiaozhuo; Bian, Xiaofang; Ji, Yongchang; Yan, Xiumin; Yang, Feng; Li, Fang

    2013-12-01

    DNA viruses often target cellular proteins to modulate host cell cycles and facilitate viral genome replication. However, whether proliferation of white spot syndrome virus (WSSV) requires regulation of the host cell cycle remains unclear. In the present study, we show that two WSSV paralogs, IE1 and WSV056, can interact with Litopenaeus vannamei retinoblastoma (Rb)-like protein (lv-RBL) through the conserved LxCxE motif. Further investigation revealed that IE1 and WSV056 could also bind to Drosophila retinoblastoma family protein 1 (RBF1) in a manner similar to how they bind to lv-RBL. Using the Drosophila RBF-E2F pathway as a model system, we demonstrated that both IE1 and WSV056 could sequester RBF1 from Drosophila E2F transcription factor 1 (E2F1) and subsequently activate E2F1 to stimulate the G1/S transition. Our findings provide the first evidence that WSSV may regulate cell cycle progression by targeting the Rb-E2F pathway.

  12. Advances in the processing of policromat images as diagnostic method to determine white spot syndrome virus in white shrimp (Litopenaeus vannamei)

    NASA Astrophysics Data System (ADS)

    Chavez-Sanchez, Cristina M.; Alvarez-Borrego, Josue; Montoya-Rodriguez, L.; Garcia-Gasca, A.; Fajer Avila, Emma J.; Pacheco-Marges, R.

    2004-10-01

    White spot syndrome (WSSV) is a viral disease which affects many crustacean species including commercial shrimps. Adequate, precise and quick methods to diagnose on time the presence of the disease in order to apply different strategies to avoid the dispersion and to reduce mortalities is necessary. Histopathology is an important diagnostic method. However, histopathology has the problem that requires time to prepare the histological slides and time to arrive to some diagnosis because this depend on the nature of the tissues, the pathogen(s) to find, the number of organisms, number of slides to analyze and the skill of the technician. This paper try to demonstrate the sensibility of one digital system of processing and recognition of images using color correlation with phase filters, to identify inclusion bodies of WSSV. Infected tissues were processed to obtain histological slides and to verify that the inclusion bodies observed were of WSV, in situ hybridization were carried out. The sensibility results of the recognition of the inclusion bodies of WSSV with the color correlation program was 86.1%. The highest percentage of recognition was in nervous system and tegument glands with 100%. The values in the stomach epithelium and heart tissue was 78.45% of recognition. Tissues with the lowest recognition values were lymphoid organ and hematopoietic tissue. It is necessary further studies to increase the sensibility and to obtain the specificity.

  13. The cDNA Sequence of Two Hemocyanin Subunits from Red Swamp Crayfish Procambarus clarkii and their Responses to White Spot Syndrome Virus Infection.

    PubMed

    Zeng, Yong

    2016-03-01

    Hemocyanin, the respiratory protein of crustaceans, participates in the innate immune defense in these organisms. We cloned two hemocyanin subunit genes (PcHc1 and PcHc2), by using a degenerate primer PCR and rapid amplification of cDNA ends (RACE) approach, from the hepatopancreas of red swamp crayfish Procambarus clarkii. The transcripts of these two subunits were only detected in the hepatopancreas by reverse transcriptase quantitative PCR (RT-qPCR) analysis. The neighbor-joining and maximum parsimony phylogenetic analyses indicated that PcHc2 associated with a clade belong to the α-type hemocyanins and PcHc1 associated with another clade belonging to the β-type hemocyanins. The data obtained from the RT-qPCR indicated that the mRNA expression levels of these subunit genes followed almost the same regulation pattern in the crayfish challenged with white spot syndrome virus (WSSV). The fluctuation of mRNA expression levels of these two subunits after the WSSV challenge indicated that both of them may participate in the antiviral immune response of crayfish. Received April 12, 2015; accepted November 22, 2015.

  14. Real-time target-specific detection of loop-mediated isothermal amplification for white spot syndrome virus using fluorescence energy transfer-based probes.

    PubMed

    Chou, Pin-Hsing; Lin, Yu-Chan; Teng, Ping-Hua; Chen, Chu-Liang; Lee, Pei-Yu

    2011-04-01

    Aiming to establish a target amplicon-specific detection system for loop-mediated isothermal amplification (LAMP), the fluorescent resonance energy transfer (FRET) probe technology was applied to develop the FRET LAMP platform. This report describes the development of the first FRET LAMP assay targeting white spot syndrome virus (WSSV) of penaeid shrimp. A successful accelerated WSSV LAMP was assembled first in a conventional oven and confirmed by gel electrophoresis and dot-blot hybridization. Subsequently, two additional FRET probes designed to target one loop region within WSSV LAMP amplicons were added to the same LAMP reaction. The reactions were carried out in a LightCycler (Roche) and significant FRET signals were detected in real time. Optimization of the reaction using plasmid DNA shortened the time for the detection of 10(2) copies of the target DNA to less than 70min. Cross reactivity was absent with WSSV-free or infectious hypodermal and hematopoietic necrosis virus-infected Penaeus vannamei samples. The performance of this system was comparable with that of a nested PCR assay from 21 WSSV-infected shrimp. Specifically detecting target amplicons and requiring no post-amplification manipulation, the novel FRET LAMP assay should allow indisputable detection of pathogens with minimized risks of amplicon contamination.

  15. SPOT Program

    NASA Technical Reports Server (NTRS)

    Smith, Jason T.; Welsh, Sam J.; Farinetti, Antonio L.; Wegner, Tim; Blakeslee, James; Deboeck, Toni F.; Dyer, Daniel; Corley, Bryan M.; Ollivierre, Jarmaine; Kramer, Leonard; Zimmerman, Patrick L.; Khatri, Reshma

    2010-01-01

    A Spacecraft Position Optimal Tracking (SPOT) program was developed to process Global Positioning System (GPS) data, sent via telemetry from a spacecraft, to generate accurate navigation estimates of the vehicle position and velocity (state vector) using a Kalman filter. This program uses the GPS onboard receiver measurements to sequentially calculate the vehicle state vectors and provide this information to ground flight controllers. It is the first real-time ground-based shuttle navigation application using onboard sensors. The program is compact, portable, self-contained, and can run on a variety of UNIX or Linux computers. The program has a modular objec-toriented design that supports application-specific plugins such as data corruption remediation pre-processing and remote graphics display. The Kalman filter is extensible to additional sensor types or force models. The Kalman filter design is also strong against data dropouts because it uses physical models from state and covariance propagation in the absence of data. The design of this program separates the functionalities of SPOT into six different executable processes. This allows for the individual processes to be connected in an a la carte manner, making the feature set and executable complexity of SPOT adaptable to the needs of the user. Also, these processes need not be executed on the same workstation. This allows for communications between SPOT processes executing on the same Local Area Network (LAN). Thus, SPOT can be executed in a distributed sense with the capability for a team of flight controllers to efficiently share the same trajectory information currently being computed by the program. SPOT is used in the Mission Control Center (MCC) for Space Shuttle Program (SSP) and International Space Station Program (ISSP) operations, and can also be used as a post -flight analysis tool. It is primarily used for situational awareness, and for contingency situations.

  16. Dark Spots

    NASA Technical Reports Server (NTRS)

    2006-01-01

    Dark spots (left) and 'fans' appear to scribble dusty hieroglyphics on top of the Martian south polar cap in two high-resolution Mars Global Surveyor, Mars Orbiter Camera images taken in southern spring. Each image is about 3-kilometers wide (2-miles).

  17. SUMO-conjugating enzyme E2 UBC9 mediates viral immediate-early protein SUMOylation in crayfish to facilitate reproduction of white spot syndrome virus.

    PubMed

    Chen, An-Jing; Gao, Lu; Wang, Xian-Wei; Zhao, Xiao-Fan; Wang, Jin-Xing

    2013-01-01

    Successful viruses have evolved superior strategies to escape host defenses or exploit host biological pathways. Most of the viral immediate-early (ie) genes are essential for viral infection and depend solely on host proteins; however, the molecular mechanisms are poorly understood. In this study, we focused on the modification of viral IE proteins by the crayfish small ubiquitin-related modifier (SUMO) and investigated the role of SUMOylation during the viral life cycle. SUMO and SUMO ubiquitin-conjugating enzyme 9 (UBC9) involved in SUMOylation were identified in red swamp crayfish (Procambarus clarkii). Both SUMO and UBC9 were upregulated in crayfish challenged with white spot syndrome virus (WSSV). Replication of WSSV genes increased in crayfish injected with recombinant SUMO or UBC9, but injection of mutant SUMO or UBC9 protein had no effect. Subsequently, we analyzed the mechanism by which crayfish SUMOylation facilitates WSSV replication. Crayfish UBC9 bound to all three WSSV IE proteins tested, and one of these IE proteins (WSV051) was covalently modified by SUMO in vitro. The expression of viral ie genes was affected and that of late genes was significantly inhibited in UBC9-silenced or SUMO-silenced crayfish, and the inhibition effect was rescued by injection of recombinant SUMO or UBC9. The results of this study demonstrate that viral IE proteins can be modified by crayfish SUMOylation, prompt the expression of viral genes, and ultimately benefit WSSV replication. Understanding of the mechanisms by which viruses exploit host components will greatly improve our knowledge of the virus-host "arms race" and contribute to the development of novel methods against virulent viruses.

  18. SUMO-Conjugating Enzyme E2 UBC9 Mediates Viral Immediate-Early Protein SUMOylation in Crayfish To Facilitate Reproduction of White Spot Syndrome Virus

    PubMed Central

    Chen, An-Jing; Gao, Lu; Wang, Xian-Wei; Zhao, Xiao-Fan

    2013-01-01

    Successful viruses have evolved superior strategies to escape host defenses or exploit host biological pathways. Most of the viral immediate-early (ie) genes are essential for viral infection and depend solely on host proteins; however, the molecular mechanisms are poorly understood. In this study, we focused on the modification of viral IE proteins by the crayfish small ubiquitin-related modifier (SUMO) and investigated the role of SUMOylation during the viral life cycle. SUMO and SUMO ubiquitin-conjugating enzyme 9 (UBC9) involved in SUMOylation were identified in red swamp crayfish (Procambarus clarkii). Both SUMO and UBC9 were upregulated in crayfish challenged with white spot syndrome virus (WSSV). Replication of WSSV genes increased in crayfish injected with recombinant SUMO or UBC9, but injection of mutant SUMO or UBC9 protein had no effect. Subsequently, we analyzed the mechanism by which crayfish SUMOylation facilitates WSSV replication. Crayfish UBC9 bound to all three WSSV IE proteins tested, and one of these IE proteins (WSV051) was covalently modified by SUMO in vitro. The expression of viral ie genes was affected and that of late genes was significantly inhibited in UBC9-silenced or SUMO-silenced crayfish, and the inhibition effect was rescued by injection of recombinant SUMO or UBC9. The results of this study demonstrate that viral IE proteins can be modified by crayfish SUMOylation, prompt the expression of viral genes, and ultimately benefit WSSV replication. Understanding of the mechanisms by which viruses exploit host components will greatly improve our knowledge of the virus-host “arms race” and contribute to the development of novel methods against virulent viruses. PMID:23097446

  19. Responses of three very large inducible GTPases to bacterial and white spot syndrome virus challenges in the giant fresh water prawn Macrobrachium rosenbergii.

    PubMed

    Huang, Ying; Jin, Min; Yin, Shaowu; Ding, Zhengfeng; Wang, Wen; Ren, Qian

    2016-04-01

    Interferons (IFNs) are cytokines secreted by cells in response to invasion by pathogens, such as viruses, bacteria, parasites, or tumor cells. Very large inducible GTPases (VLIG) are the latest IFN-inducible GTPase family to be discovered and are the largest known GTPases of any species. However, VLIG proteins from invertebrates have yet to be characterized. In this study, three forms of VLIGs designated as MrVLIG1, MrVLIG2, and MrVLIG3 were cloned from the giant fresh water prawn Macrobrachium rosenbergii. MrVLIG1 has a 5445 bp open reading frame (ORF) encoding an 1814-amino acid protein. The complete nucleotide sequence of MrVLIG2 cDNA is 7055 bp long consisting of a 5757 bp ORF encoding a protein with 1918 amino acids. The full length of the MrVLIG3 gene consists of 5511 bp with a 3909 bp ORF encoding a peptide with 1302 amino acids. BLASTP and phylogenetic tree analyses showed that the three MrVLIGs are clustered into one subgroup and, together with other vertebrate VLIGs, into a branch. Tissue distribution analysis indicated that the mRNAs of the three MrVLIGs were widely expressed in almost all detected tissues, including the hemocytes, heart, hepatopancreas, gills, stomach, and intestine, with the highest expression in the hepatopancreas. They were also detected in the intestine but with relatively low expression levels. Quantitative real-time RT-PCR analysis showed that the mRNA transcripts of the MrVLIGs in the hepatopancreas were significantly expressed at various time points after infection with Vibrio parahaemolyticus and white spot syndrome virus. In summary, the three isoforms of VLIG genes participate in the innate immune response of the shrimps to bacterial and viral infections.

  20. An immersion of Gracilaria tenuistipitata extract improves the immunity and survival of white shrimp Litopenaeus vannamei challenged with white spot syndrome virus.

    PubMed

    Lin, Yong-Chin; Yeh, Su-Tuen; Li, Chang-Che; Chen, Li-Li; Cheng, Ann-Chang; Chen, Jiann-Chu

    2011-12-01

    The innate immunity and resistance against white spot syndrome virus (WSSV) in white shrimp Litopenaeus vannamei which received the Gracilaria tenuistipitata extract were examined. Shrimp immersed in seawater containing the extract at 0 (control), 400 and 600 mg L(-1) for 3 h were challenged with WSSV at 2 × 10(4) copies shrimp(-1). Shrimp not exposed to the extract and not received WSSV challenge served as unchallenged control. The survival rate of shrimp immersed in 400 mg L(-1) or 600 mg L(-1) extract was significantly higher than that of challenged control shrimp over 24-120 h. The haemocyte count, phenoloxidase activity, respiratory burst, superoxide dismutase activity, and lysozyme activity of shrimp immersed in 600 mg L(-1) extract were significantly higher than those of unchallenged control shrimp at 6, 6, 6, 6, and 6-24 h post-challenge. In another experiment, shrimp which had received 3 h immersion of 0, 400, 600 mg L(-1) extract were challenged with WSSV. The shrimp were then received a booster (3 h immersion in the same dose of the extract), and the immune parameters were examined at 12-120 h post-challenge. The immune parameters of shrimp immersed in 600 mg L(-1) extract, and then received a booster at 9, 21, and 45 h were significantly higher than those of unchallenged control shrimp at 12-48 h post-challenge. In conclusion, shrimp which had received the extract exhibited protection against WSSV as evidenced by the higher survival rate and higher values of immune parameters. Shrimp which had received the extract and infected by WSSV showed improved immunity when they received a booster at 9, 21, and 45 h post-WSSV challenge. The extract treatment caused less decrease in PO activity, and showed better performance of lysozyme activity and antioxidant response in WSSV-infected shrimp.

  1. Blocking the large extracellular loop (LEL) domain of FcTetraspanin-3 could inhibit the infection of white spot syndrome virus (WSSV) in Chinese shrimp, Fenneropenaeus chinensis.

    PubMed

    Gui, Lang; Wang, Bing; Li, Fu-Hua; Sun, Yu-Miao; Luo, Zhan; Xiang, Jian-Hai

    2012-06-01

    Tetraspanins belong to the transmembrane 4 superfamily (TM(4)SF), which span the cell membrane 4 times and act as bridges or connectors. Increasing evidences have shown that tetraspanins play important role in virus infection. The large extracellular loop (LEL) of a tetraspanin is considered as a possible target of some virus. Tetraspanins are widely found in invertebrates, but the functional roles of most invertebrate tetraspanins have remained unknown. Recently, a tetraspanin, called FcTetraspanin-3, was cloned from the cDNA library of Chinese shrimp, Fenneropenaeus chinensis. The FcTetraspanin-3 constitutive expression in all examined tissues and the expression of the gene were highly induced in hepatopancreas, lymphoid organ and intestine by white spot syndrome virus (WSSV) challenge. In this study, we expressed and purified the recombinant peptide containing the LEL domain of FcTetraspanin-3, and produced the anti-LEL polyclone antibody. The expression of FcTetraspanin-3 was observed by real-time PCR and Western blot. Also, the localization of FcTetraspanin-3-positive cells in intestine and hepatopancreas were revealed by immunofluorescence. The results of anti-LEL antibody blocking experiments shown that the antibody can significantly reduce the mortality of shrimp challenged by WSSV. Additionally, dsRNA interference was utilized to examine the functional role of FcTetraspanin-3 in response to WSSV infection, and a sensible decrease of the viral copy number in the tetraspanin knockdown shrimp. These results suggested the blocking of LEL domain of FcTetraspanin-3 could inhibit the infection of WSSV. FcTetraspanin-3 might play an important role in response to WSSV infection, and the LEL domain of FcTetraspanin-3 might mediate the entry of WSSV.

  2. Dietary administration of a Gracilaria tenuistipitata extract enhances the immune response and resistance against Vibrio alginolyticus and white spot syndrome virus in the white shrimp Litopenaeus vannamei.

    PubMed

    Sirirustananun, Nuttarin; Chen, Jiann-Chu; Lin, Yong-Chin; Yeh, Su-Tuen; Liou, Chyng-Hwa; Chen, Li-Li; Sim, Su Sing; Chiew, Siau Li

    2011-12-01

    The haemogram, phenoloxidase (PO) activity, respiratory bursts (RBs), superoxide dismutase (SOD) activity, glutathione peroxidase (GPx) activity, lysozyme activity, and the mitotic index of haematopoietic tissue (HPT) were examined after the white shrimp Litopenaeus vannamei had been fed diets containing the hot-water extract of Gracilaria tenuistipitata at 0 (control), 0.5, 1.0, and 2.0 g kg(-1) for 7-35 days. Results indicated that these parameters directly increased with the amount of extract and time, but slightly decreased after 35 days. RBs, SOD activity, and GPx activity reached the highest levels after 14 days, whereas PO and lysozyme activities reached the highest levels after 28 days. In a separate experiment, white shrimp L. vannamei, which had been fed diets containing the extract for 14 days, were challenged with Vibrio alginolyticus at 2 × 10(6) cfu shrimp(-1) and white spot syndrome virus (WSSV) at 1 × 10(3) copies shrimp(-1), and then placed in seawater. The survival rate of shrimp fed the extract-containing diets was significantly higher than that of shrimp fed the control diet at 72-144 h post-challenge. We concluded that dietary administration of the G. tenuistipitata extract at ≤1.0 g kg(-1) could enhance the innate immunity within 14 days as evidenced by the increases in immune parameters and mitotic index of HPT in shrimp and their enhanced resistance against V. alginolyticus and WSSV infections. Shrimp fed the extract-containing diets showed a higher and continuous increase in the humoral response indicating its persistent role in innate immunity.

  3. Expression Profile of Penaeus monodon Ubiquitin Conjugating Enzyme (PmUbc) at Protein Level in White spot syndrome virus Challenged Shrimp.

    PubMed

    Keezhedath, Jeena; Kurcheti, Pani Prasad; Pathan, Mujahid Khan; Babu, Gireesh P; Tripathi, Gayatri; Sudhagar, Arun; Rao, Srinivas P

    2013-06-01

    White spot syndrome virus (WSSV) is one of the major pathogens in shrimp aquaculture. Four proteins of WSSV are predicted to encode a RING H2 domain, which in presence of ubiquitin conjugating enzyme (E2) in shrimps can function as viral E3 ligase and modulate the host ubiquitin proteasome pathway. Modulation of host ubiquitin proteasome pathway by viral proteins is implicated in viral pathogenesis. In the present study, expression profile of Penaeus monodon Ubiquitin conjugating enzyme (PmUbc) was studied at protein level in WSSV challenged shrimp. A time point analysis of the expression of PmUbc was carried out at 0, 3, 6, 12, 24, 48 and 72 h post WSSV challenge in P. monodon. Recombinant PmUbc (rPmUbc) was produced in prokaryotic expression vector, BL21 (DE3) pLys S. The PmUbc expression pattern was studied by ELISA with rPmUbc antibodies raised in rabbit. A significant increase in PmUbc expression at 24 h post infection (hpi) was observed followed by a decline till 72 hpi. Since the up-regulation and a tremendous decline of PmUbc protein expression was observed at 24 and in 72 hpi respectively in ELISA, it can be speculated that these proteins might interact with host ubiquitination pathway for viral pathogenesis. Many findings have shown that viral infection can up-regulate expression of ubiquitin and that the ubiquitin system plays a key role in the course of viral infection. The present study reveals the expression patterns of PmUbc at protein level in WSSV infected P. monodon. However, further studies are to be carried out to unfold the molecular mechanism of interaction between host and virus to devise efficient control strategies for this major culprit in shrimp culture industry.

  4. Activating Transcription Factor 4 and X Box Binding Protein 1 of Litopenaeus vannamei Transcriptional Regulated White Spot Syndrome Virus Genes Wsv023 and Wsv083

    PubMed Central

    Li, Xiao-Yun; Pang, Li-Ran; Chen, Yong-Gui; Weng, Shao-Ping; Yue, Hai-Tao; Zhang, Ze-Zhi; Chen, Yi-Hong; He, Jian-Guo

    2013-01-01

    In response to endoplasmic reticulum (ER) stress, the signaling pathway termed unfolded protein response (UPR) is activated. To investigate the role of UPR in Litopenaeus vannamei immunity, the activating transcription factor 4 (designated as LvATF4) which belonged to a branch of the UPR, the [protein kinase RNA (PKR)-like ER kinase, (PERK)]-[eukaryotic initiation factor 2 subunit alpha (eIF2α)] pathway, was identified and characterized. The full-length cDNA of LvATF4 was 1972 bp long, with an open reading frame of 1299 bp long that encoded a 432 amino acid protein. LvATF4 was highly expressed in gills, intestines and stomach. For the white spot syndrome virus (WSSV) challenge, LvATF4 was upregulated in the gills after 3 hpi and increased by 1.9-fold (96 hpi) compared to the mock-treated group. The LvATF4 knock-down by RNA interference resulted in a lower cumulative mortality of L. vannamei under WSSV infection. Reporter gene assays show that LvATF4 could upregulate the expression of the WSSV gene wsv023 based on the activating transcription factor/cyclic adenosine 3′, 5′-monophosphate response element (ATF/CRE). Another transcription factor of L. vannamei, X box binding protein 1 (designated as LvXBP1), has a significant function in [inositol-requiring enzyme-1(IRE1) – (XBP1)] pathway. This transcription factor upregulated the expression of the WSSV gene wsv083 based on the UPR element (UPRE). These results suggest that in L. vannamei UPR signaling pathway transcription factors are important for WSSV and might facilitate WSSV infection. PMID:23638122

  5. Marine derived compounds as binders of the White spot syndrome virus VP28 envelope protein: In silico insights from molecular dynamics and binding free energy calculations.

    PubMed

    Sivakumar, K C; Sajeevan, T P; Bright Singh, I S

    2016-10-01

    White spot syndrome virus (WSSV) remains as one of the most dreadful pathogen of the shrimp aquaculture industry owing to its high virulence. The cumulative mortality reaches up to 100% within in 2-10days in a shrimp farm. Currently, no chemotherapeutics are available to control WSSV. The viral envelope protein, VP28, located on the surface of the virus particle acts as a vital virulence factor in the initial phases of inherent WSSV infection in shrimp. Hence, inhibition of envelope protein VP28 could be a novel way to deal with infection by inhibiting its interaction in the endocytic pathway. In this direction, a timely attempt was made to recognize a potential drug candidate of marine origin against WSSV using VP28 as a target by employing in silico docking and molecular dynamic simulations. A virtual library of 388 marine bioactive compounds was extracted from reports published in Marine Drugs. The top ranking compounds from docking studies were chosen from the flexible docking based on the binding affinities (ΔGb). In addition, the MD simulation and binding free energy analysis were implemented to validate and capture intermolecular interactions. The results suggested that the two compounds obtained a negative binding free energy with -40.453kJ/mol and -31.031kJ/mol for compounds with IDs 30797199 and 144162 respectively. The RMSD curve indicated that 30797199 moves into the hydrophobic core, while the position of 144162 atoms changes abruptly during simulation and is mostly stabilized by water bridges. The shift in RMSD values of VP28 corresponding to ligand RMSD gives an insight into the ligand induced conformational changes in the protein. This study is first of its kind to elucidate the explicit binding of chemical inhibitor to WSSV major structural protein VP28.

  6. The novel white spot syndrome virus-induced gene, PmERP15, encodes an ER stress-responsive protein in black tiger shrimp, Penaeus monodon.

    PubMed

    Leu, Jiann-Horng; Liu, Kuan-Fu; Chen, Kuan-Yu; Chen, Shu-Hwa; Wang, Yu-Bin; Lin, Chung-Yen; Lo, Chu-Fang

    2015-04-01

    By microarray screening, we identified a white spot syndrome virus (WSSV)-strongly induced novel gene in gills of Penaeus monodon. The gene, PmERP15, encodes a putative transmembrane protein of 15 kDa, which only showed some degree of similarity (54-59%) to several unknown insect proteins, but had no hits to shrimp proteins. RT-PCR showed that PmERP15 was highly expressed in the hemocytes, heart and lymphoid organs, and that WSSV-induced strong expression of PmERP15 was evident in all tissues examined. Western blot analysis likewise showed that WSSV strongly up-regulated PmERP15 protein levels. In WSSV-infected hemocytes, immunofluorescence staining showed that PmERP15 protein was colocalized with an ER enzyme, protein disulfide isomerase, and in Sf9 insect cells, PmERP15-EGFP fusion protein colocalized with ER -Tracker™ Red dye as well. GRP78, an ER stress marker, was found to be up-regulated in WSSV-infected P. monodon, and both PmERP15 and GRP78 were up-regulated in shrimp injected with ER stress inducers tunicamycin and dithiothreitol. Silencing experiments showed that although PmERP15 dsRNA-injected shrimp succumbed to WSSV infection more rapidly, the WSSV copy number had no significant changes. These results suggest that PmERP15 is an ER stress-induced, ER resident protein, and its induction in WSSV-infected shrimp is caused by the ER stress triggered by WSSV infection. Furthermore, although PmERP15 has no role in WSSV multiplication, its presence is essential for the survival of WSSV-infected shrimp.

  7. Protection of Penaeus monodon against white spot syndrome by continuous oral administration of a low concentration of Bacillus subtilis spores expressing the VP28 antigen.

    PubMed

    Pham, K-C; Tran, H T T; Van Doan, C; Le, P H; Van Nguyen, A T; Nguyen, H A; Hong, H A; Cutting, S M; Phan, T-N

    2017-03-01

    In this study, Bacillus subtilis spores expressing a chimeric protein, CotB-VP28, were used as a probiotic vaccine to protect black tiger shrimps (Penaeus monodon) against white spot syndrome virus (WSSV) infection. Oral administration of pellets coated with CotB-VP28 spores (at ≥1 × 10(9 ) CFU per g pellet) to shrimps induced immune-relating phenoloxydase activity (PO) in shrimps after 14 days of feeding (prior challenge) and at day 3 post challenge (1·26 and 1·70 fold increase respectively). A 75% protection rate was obtained by continuous feeding of the spore-coated pellets at ≥1 × 10(9 ) CFU per g for 14 days prior to WSSV challenge and during all the postchallenge period. Even when the amount of CotB-VP28 spores in feed pellets was reduced down to ≥5 × 10(7)  CFU per g and ≥1 × 10(6)  CFU per g, relatively high protection rates of 70 and 67·5%, respectively, were still obtained. By contrast, feeding pellets without spores (untreated group) and with naked spores (PY79 group) at ≥1 × 10(9)  CFU per g could not protect shrimps against WSSV. These data suggest that supplementation of CotB-VP28 spores at low dose of ≥1 × 10(6)  CFU per g could be effective as a prophylactic treatment of WSS for black tiger shrimps.

  8. The shrimp IKK-NF-κB signaling pathway regulates antimicrobial peptide expression and may be subverted by white spot syndrome virus to facilitate viral gene expression.

    PubMed

    Wang, Pei-Hui; Gu, Zhi-Hua; Wan, Ding-Hui; Liu, Bo-Du; Huang, Xian-De; Weng, Shao-Ping; Yu, Xiao-Qiang; He, Jian-Guo

    2013-09-01

    The IκB kinases IKKα and IKKβ and the IKK-related kinases TANK-binding kinase 1 (TBK1) and IKKε are the master regulators of the NF-κB signaling pathway. Although this pathway has been extensively studied in mammals, less attention has been paid in crustaceans, which have significant economic value. Here, we report the cloning and functional studies of two IKK homologs, LvIKKβ and LvIKKε, from Pacific white shrimp, Litopenaeus vannamei. LvIKKβ and LvIKKε mRNAs are widely expressed in different tissues and are responsive to white spot syndrome virus (WSSV) infection. When overexpressed in Drosophila S2 cells, LvIKKβ but not LvIKKε activates the promoters of NF-κB pathway-controlled antimicrobial peptide genes (AMPs), such as the Penaeidins (PENs). In HEK 293T cells, both LvIKKβ and LvIKKε activate an NF-κB reporter. The silencing of LvIKKβ or LvIKKε using double-stranded RNA (dsRNA)-mediated RNA interference (RNAi) decreases the expression of L. vannamei AMPs, including PENs, lysozyme and crustins. Intriguingly, LvIKKβ- or LvIKKε-silenced L. vannamei are resistant to WSSV infection. We hypothesized that successful infection with WSSV requires the activation of the IKK-NF-κB signaling pathway to modulate viral gene expression. We constructed luciferase reporters for 147 WSSV genes. By screening, we found that the WSV051, WSV059, WSV069, WSV083, WSV090, WSV107, WSV244, WSV303, WSV371 and WSV445 promoters can be activated by LvIKKβ or LvIKKε in Drosophila S2 cells. Taken together, our results reveal that LvIKKβ and LvIKKε may participate in the regulation of shrimp AMPs and that WSSV may subvert the L. vannamei IKK-NF-κB signaling pathway to facilitate viral gene expression.

  9. Characterization of microRNAs by deep sequencing in red claw crayfish Cherax quadricarinatus haematopoietic tissue cells after white spot syndrome virus infection.

    PubMed

    Zhao, Meng-Ru; Meng, Chuang; Xie, Xiao-Lu; Li, Cheng-Hua; Liu, Hai-Peng

    2016-12-01

    White spot syndrome virus (WSSV) is one of the most prevalent and widespread viruses in both shrimp and crayfish aquaculture. MicroRNAs (miRNAs) are crucial post-transcriptional regulators and play critical roles in cell differentiation and proliferation, apoptosis, signal transduction and immunity. In this study, miRNA expression profiles were identified via deep sequencing in red claw crayfish Cherax quadricarinatus haematopoietic tissue (Hpt) cell cultures infected with WSSV at both early (i.e., 1 hpi) and late (i.e., 12 hpi) infection stages. The results showed that 2 known miRNAs, namely, miR-7 and miR-184 play key roles in immunity. Meanwhile, 106 novel miRNA candidates were predicted by software in these combined miRNA transcriptomes. Compared with two control groups, 36 miRNAs showed significantly different expression levels after WSSV challenge. Furthermore, 10 differentially expressed miRNAs in WSSV-exposed Hpt cells were randomly selected for expression analysis by quantitative real-time RT-PCR. Consistent with the expression profiles identified by deep sequencing, RT-PCR showed a significant increase or decrease in miRNA expression in Hpt cells after WSSV infection. Prediction of targets of miRNAs such as miR-7, cqu-miR-52, cqu-miR-126 and cqu-miR-141 revealed that their target genes have diverse biological roles, including not only immunity but also transcriptional regulation, energy metabolism, cell communication, cell differentiation, cell death, autophagy, endocytosis and apoptosis. These results provide insight into the molecular mechanism of WSSV infection and highlight the function of miRNAs in the regulation of the immune response against WSSV infection in crustaceans.

  10. Immune responses of whiteleg shrimp, Litopenaeus vannamei (Boone, 1931), to bacterially expressed dsRNA specific to VP28 gene of white spot syndrome virus.

    PubMed

    Taju, G; Madan, N; Abdul Majeed, S; Kumar, T Raj; Thamizhvanan, S; Otta, S K; Sahul Hameed, A S

    2015-05-01

    In this study, dsRNA specific to VP28 gene of white spot syndrome virus (WSSV) of shrimp was synthesized in Escherichia coli in large scale and studied the immune response of shrimp to dsRNA-VP28. The haematological parameters such as clotting time and total haemocytes counts, and immunological parameters such as prophenoloxidase (proPO), superoxide dismutase (SOD), superoxide anion (SOA) and malondialdehyde content, as well as the mRNA expression of ten immune-related genes were examined to estimate the effect of dsRNA-VP28 on the innate immunity of Litopenaeus vannamei. The activities of proPO, SOA and SOD significantly increased in haemocyte after dsRNA-VP28 treatment, whereas MDA content did not change significantly. Among the ten immune-related genes examined, only the mRNA expression of proPO, cMnSOD, haemocyanin, crustin, BGBP, lipopolysaccharides (LPs), lectin and lysozyme in haemocytes, gill and hepatopancreas of L. vannamei, was significantly upregulated at 12 h after dsRNA-VP28 treatment, while no significant expression changes were observed in Toll receptor and tumour receptor genes. The increase of proPO and SOD activities, and SOA level and mRNA expression level of proPO, cMnSOD, haemocyanin, crustin, BGBP, LPs, lectin and lysozyme after dsRNA-VP28 stimulation indicate that these immune-related genes were involved in dsRNA-VP28-induced innate immunity in shrimp.

  11. Analysis of variable genomic loci in white spot syndrome virus to predict its origins in Procambarus clarkii crayfish farmed in China.

    PubMed

    Zeng, Wei; Zeng, Yong; Fei, Rong-Mei; Zeng, Ling-Bing; Wei, Kai-Jin

    2011-09-09

    Variable genomic loci were examined in 4 white spot syndrome virus (WSSV) isolates (08HB, 09HB, 08JS and 09JS) from Procambarus clarkii crayfish collected from Jiangsu and Hubei Provinces in China in 2008 and 2009. In ORF75, sequence variation detected in the 4 isolates, as well as in isolates sequenced previously, suggested that WSSV might have segregated into 2 lineages since first emerging as a serious pathogen of farmed shrimp in East Asia in the early-mid 1990s, with one lineage remaining in East Asia and the other separating to South Asia. In ORF23/24, deletions of 9.31, 10.97, or 11.09 kb were evident compared to a reference isolate from Taiwan (WSSV-TW), and, in ORF14/15, deletions of 5.14 or 5.95 kb were evident compared to a reference isolate from Thailand with the largest genome size (TH-96-II). With respect to these genome characteristics, the crayfish isolates 08HB, 09HB and 08JS were similar to WSSV-TW and the isolate 09JS was similar to a reference isolate from China (WSSV-CN). In addition to these loci, sequence variation was evident in ORF94 and ORF125 that might be useful for differentiating isolates and in epidemiological tracing of WSSV spread in crayfish farmed in China. However, as all 4 crayfish isolates possessed a Homologous Region 9 sequence identical to isolate WSSV-TW and another Thailand isolate (WSSV-TH), and as their transposase sequence was identical to isolates WSSV-CN and WSSV-TH, these 2 loci were not useful in predicting their origins.

  12. Anti-melanization mechanism of the white spot syndrome viral protein, WSSV453, via interaction with shrimp proPO-activating enzyme, PmproPPAE2.

    PubMed

    Sutthangkul, Jantiwan-; Amparyup, Piti-; Eum, Jai Hoon; Strand, Michael R; Tassanakajon, Anchalee

    2017-01-28

    Inhibition of the host melanization reaction, activated by the prophenoloxidase activating (proPO) system, is one of the crucial evasion strategies of pathogens. Recently, the shrimp pathogen, white spot syndrome virus (WSSV), was found to inhibit melanization in the shrimp, Penaeus monodon. The viral protein WSSV453 was previously shown to interact with PO-activating enzyme 2 (PmPPAE2) and reported to be involved in suppressing the shrimp melanization response after WSSV infection. Here, we characterized how WSSV453 inhibits melanization. WSSV453 is a non-structural viral protein, which was first detected in shrimp hemocytes at 6 hours post infection (hpi) by WSSV and in shrimp plasma at 24 hpi. We produced recombinant proteins for three components of the P. monodon, proPO system: PmproPPAE2, PmproPO1 and PmproPO2. Functional assays showed that active PmPPAE2 processed PmproPO1 and 2 to produce functional PO. Incubation of WSSV453 with PmproPPAE2 dose-dependently reduced PmPPAE2 activity toward PmPO1 or PmPO2. In contrast, WSSV453 had no effect on activated PmPPAE2. The addition of active PmPPAE2 to WSSV-infected shrimp plasma at day 2 post-infection also rescued PO activity. Taken together, these results indicate that the anti-melanization activity of WSSV is due to WSSV453, which interacts with PmproPPAE2 and interferes its activation to active PmPPAE2.

  13. The c-Fos and c-Jun from Litopenaeus vannamei play opposite roles in Vibrio parahaemolyticus and white spot syndrome virus infection.

    PubMed

    Li, Chaozheng; Li, Haoyang; Wang, Sheng; Song, Xuan; Zhang, Zijian; Qian, Zhe; Zuo, Hongliang; Xu, Xiaopeng; Weng, Shaoping; He, Jianguo

    2015-09-01

    Growing evidence indicates that activator protein-1 (AP-1) plays a major role in stimulating the transcription of immune effector molecules in cellular response to an incredible array of stimuli, including growth factors, cytokines, cellular stresses and bacterial and viral infection. Here, we reported the isolation and characterization of a cDNA from Litopenaeus vannamei encoding the full-length c-Fos protein (named as Lvc-Fos). The predicted amino acid sequences of Lvc-Fos contained a basic-leucine zipper (bZIP) domain, which was characteristic of members of the AP-1 family. Immunoprecipitation and native-PAGE assays determined that Lvc-Fos could interact with the Lvc-Jun, a homolog of c-Jun family in L. vannamei, in a heterodimer manner. Further investigation demonstrated that Lvc-Fos and Lvc-Jun were expressed in all tested tissues and located in the nucleus. Real-time RT-PCR analysis showed both Lvc-Fos and Lvc-Jun in gills were up-regulated during Vibrio parahaemolyticus and white spot syndrome virus (WSSV) challenges. In addition, reporter gene assays indicated Lvc-Fos and Lvc-Jun could activate the expression of antimicrobial peptides (AMPs) of Drosophila and shrimp, as well as WSSV immediate early (IE) genes wsv069 and wsv249, in a different manner. Knockdown of Lvc-Fos or Lvc-Jun by RNA interference (RNAi) resulted in higher mortalities of L. vannamei after infection with V. parahaemolyticus, suggesting that Lvc-Fos and Lvc-Jun might play protective roles in bacterial infection. However, silencing of Lvc-Fos or Lvc-Jun in shrimp caused lower mortalities and virus loads under WSSV infection, suggesting that Lvc-Fos and Lvc-Jun could be engaged for WSSV replication and pathogenesis. In conclusion, our results provided experimental evidence and novel insight into the roles of L. vannamei AP-1 in bacterial and viral infection.

  14. Crystal Structures of Major Envelope Proteins VP26 and VP28 from White Spot Syndrome Virus Shed Light on Their Evolutionary Relationship

    SciTech Connect

    Tang,X.; Wu, J.; Sivaraman, J.; Hew, C.

    2007-01-01

    White spot syndrome virus (WSSV) is a virulent pathogen known to infect various crustaceans. It has bacilliform morphology with a tail-like appendage at one end. The envelope consists of four major proteins. Envelope structural proteins play a crucial role in viral infection and are believed to be the first molecules to interact with the host. Here, we report the localization and crystal structure of major envelope proteins VP26 and VP28 from WSSV at resolutions of 2.2 and 2.0 {angstrom}, respectively. These two proteins alone account for approximately 60% of the envelope, and their structures represent the first two structural envelope proteins of WSSV. Structural comparisons among VP26, VP28, and other viral proteins reveal an evolutionary relationship between WSSV envelope proteins and structural proteins from other viruses. Both proteins adopt {beta}-barrel architecture with a protruding N-terminal region. We have investigated the localization of VP26 and VP28 using immunoelectron microscopy. This study suggests that VP26 and VP28 are located on the outer surface of the virus and are observed as a surface protrusion in the WSSV envelope, and this is the first convincing observation for VP26. Based on our studies combined with the literature, we speculate that the predicted N-terminal transmembrane region of VP26 and VP28 may anchor on the viral envelope membrane, making the core {beta}-barrel protrude outside the envelope, possibly to interact with the host receptor or to fuse with the host cell membrane for effective transfer of the viral infection. Furthermore, it is tempting to extend this host interaction mode to other structural viral proteins of similar structures. Our finding has the potential to extend further toward drug and vaccine development against WSSV.

  15. Defrosting Spots

    NASA Technical Reports Server (NTRS)

    2005-01-01

    3 October 2005 This Mars Global Surveyor (MGS) Mars Orbiter Camera (MOC) image shows dark, defrosting spots formed on a polygon-cracked plain in the south polar region of Mars. The surface was covered with carbon dioxide frost during the previous winter. In spring, the material begins to sublime away, creating a pattern of dark spots that sometimes have wind streaks emanating from them, as wind carries away or erodes the frost.

    Location near: 87.2oS, 28.4oW Image width: width: 3 km (1.9 mi) Illumination from: upper left Season: Southern Spring

  16. Simultaneous and rapid detection of white spot syndrome virus and yellow head virus infection in shrimp with a dual immunochromatographic strip test.

    PubMed

    Sithigorngul, Paisarn; Rukpratanporn, Sombat; Chaivisuthangkura, Parin; Sridulyakul, Pattarin; Longyant, Siwaporn

    2011-04-01

    A strip test for the dual detection of white spot syndrome virus (WSSV) and yellow head virus (YHV) was developed using monoclonal antibodies (MAbs) specific to the WSSV major envelope protein VP28 (W1 and W30) and the YHV nucleocapsid protein p20 (Y19 and Y21). The MAbs W30 and Y19 were conjugated with colloidal gold and sprayed onto a glass fiber pad that was placed adjacent to a sample chamber. The MAbs W1 and Y21 and the goat anti-mouse immunoglobulin G (GAM) antibody were sprayed onto a nitrocellulose membrane in strips at positions designated W, Y and C, respectively. These test strips were placed in plastic cases and stored desiccated in a plastic bag. The test strips were assessed for their ability to detect WSSV and YHV simultaneously using pleopods sampled from shrimp. A pleopod homogenate in application buffer 100μl was applied to the sample chamber to flow through the nitrocellulose membrane strip, and antibody-protein complexes could be observed within 15min. In sample from shrimp infected with WSSV and/or YHV, viral protein bound to the colloidal gold-conjugated MAbs. These complexes were captured by the MAbs at the W and/or Y test lines, resulting in the appearance of reddish-purple coloured bands. Any unbound colloidal gold-conjugated MAbs migrated pass the W and Y lines would be captured by the GAM antibody, forming a band at position C. When samples not containing WSSV and YHV proteins or containing viral proteins at below the detection limit of the test, only the band at position C was observed. The sensitivity of the test was comparable to dot blot tests using single MAbs, and ∼500-fold less sensitive than a 1-step PCR test for WSSV and 1000-fold less sensitive than an RT-PCR test for YHV. Despite this lower sensitivity, the dual strip test has advantages in speed and simplicity in not requiring sophisticated equipment or specialized skills. The ability to co-detect WSSV and YHV provides simultaneously cost savings.

  17. SUMO and SUMO-Conjugating Enzyme E2 UBC9 Are Involved in White Spot Syndrome Virus Infection in Fenneropenaeus chinensis

    PubMed Central

    Tang, Xiaoqian; Li, Wei; Xing, Jing; Sheng, Xiuzhen; Zhan, Wenbin

    2016-01-01

    In previous work, small ubiquitin-like modifier (SUMO) in hemocytes of Chinese shrimp Fenneropenaeus chinensis was found to be up-regulated post-white spot syndrome virus (WSSV) infection using proteomic approach. However, the role of SUMO in viral infection is still unclear. In the present work, full length cDNAs of SUMO (FcSUMO) and SUMO-conjugating enzyme E2 UBC9 (FcUBC9) were cloned from F. chinensis using rapid amplification of cDNA ends approach. The open reading frame (ORF) of FcSUMO encoded a 93 amino acids peptide with the predicted molecular weight (M.W) of 10.55 kDa, and the UBC9 ORF encoded a 160 amino acids peptide with the predicted M.W of 18.35 kDa. By quantitative real-time RT-PCR, higher mRNA transcription levels of FcSUMO and FcUBC9 were detected in hemocytes and ovary of F. chinensis, and the two genes were significantly up-regulated post WSSV infection. Subsequently, the recombinant proteins of FcSUMO and FcUBC9 were expressed in Escherichia coli BL21 (DE3), and employed as immunogens for the production of polyclonal antibody (PAb). Indirect immunofluorescence assay revealed that the FcSUMO and UBC9 proteins were mainly located in the hemocytes nuclei. By western blotting, a 13.5 kDa protein and a 18.7 kDa protein in hemocytes were recognized by the PAb against SUMO or UBC9 respectively. Furthermore, gene silencing of FcSUMO and FcUBC9 were performed using RNA interference, and the results showed that the number of WSSV copies and the viral gene expressions were inhibited by knockdown of either SUMO or UBC9, and the mortalities of shrimp were also reduced. These results indicated that FcSUMO and FcUBC9 played important roles in WSSV infection. PMID:26927328

  18. Ferritin administration effectively enhances immunity, physiological responses, and survival of Pacific white shrimp (Litopenaeus vannamei) challenged with white spot syndrome virus.

    PubMed

    Ruan, Yuan-Hwa; Kuo, Ching-Ming; Lo, Chu-Fang; Lee, Min-Hsien; Lian, Juang-Lin; Hsieh, Shu-Ling

    2010-04-01

    We examined the physiological (hemolymph glucose, lactate, and lipid) and innate non-specific immune responses (total hemocyte count (THC), phenoloxidase (PO) activity, respiratory bursts (release of superoxide anion, O(2)(-)) and superoxide dismutase (SOD) activity) to white spot syndrome virus (WSSV) in white shrimp (Litopenaeus vannamei) that were individually injected with 0.1, 0.5, and 1 ng g(-1) ferritin. Results showed that the THC, PO activity, and respiratory bursts of L. vannamei obviously increased (p < 0.05) 12 h after being injected with any dose of ferritin. However, the THC, PO activity, and respiratory bursts of L. vannamei that had received 0.5 and 1 ng g(-1) ferritin were significant higher than those of the other groups at 36-60, 60-72, and 36-60 h, respectively. SOD activities of L. vannamei 12 h after receiving 0.1, 0.5, and 1 ng g(-1) ferritin were significantly higher than those receiving saline. L. vannamei injected with ferritin at any dose maintained lower glucose, lactate, and lipid levels in response to WSSV challenge after 12-36, 24-48, and 36-60 h, respectively. The survival of shrimp that had received 0.5 and 1 ng g(-1) ferritin was significantly higher than that of shrimp that received saline and of control shrimp after 72 h. The ferritin messenger RNA transcripts of shrimp that had received 0.5 and 1 ng g(-1) ferritin were significantly higher than that of shrimp that received saline after 36 h. It was, therefore, concluded that the immune ability and resistance against WSSV infection increased in L. vannamei that had received > 0.5 ng g(-1) ferritin. Ferritin does play important roles in the innate immunity of the white shrimp. We observed higher SOD activities of L. vannamei that had received 0.1, 0.5, and 1 ng ferritin after 12 h than those that had received only saline (control), and the high SOD expression remained at the same levels even after 72 h of treatment.

  19. Influence of Agathi grandiflora active principles inhibit viral multiplication and stimulate immune system in Indian white shrimp Fenneropenaeus indicus against white spot syndrome virus infection.

    PubMed

    Bindhu, Francis; Velmurugan, Subramanian; Donio, Mariathason Birdilla Selva; Michaelbabu, Mariavincent; Citarasu, Thavasimuthu

    2014-12-01

    Five herbs including Adathoda vasica, Agathi grandiflora, Leucas aspera, Psoralea corylifolia, and Quercus infectoria were selected to screen the antiviral and immunostimulant activity against white spot syndrome virus (WSSV) and Vibrio harveyi respectively using different organic polar and non-polar solvents. Based on the initial screening results, ethyl acetate and methanolic extracts of A. grandiflora had strong antiviral and immunostimulant activities. Those extracts incubated with WSSV injected Fenneropenaeus indicus got only 20% mortality and no PCR positive signals were seen in two step PCR amplification. The methanolic extracts of A. grandiflora were further purified through silica column chromatography and the fractions screened again for antiviral and immunostimulant activity. The secondary screening results revealed that, the fractions of F5 to F7 had effectively controlled the WSSV multiplication and V. harveyi growth. The pooled fractions (F5 to F7) was structurally characterized by gas chromatograph-mass spectrometry (GC-MS) analysis and few compounds were identified including 3,7.11,15-Tetramethyl-2-Hexane-1-ol, pytol and 1,2-Benzenedicarboxylic acid, diisooctyl ester. The pooled fractions were mixed with the basal feed ingredients at the concentration of 100 (D-1), 200 (D-2), 300 (D-3) and 400 (D-4) mg kg(-1) and the diets fed to the F. indicus (9.0 ± 0.5 g) for 30 days. After the completion of feeding trail, they were challenged with virulent WSSV and studied the cumulative mortality, molecular diagnosis by quantitative real time PCR (qRT-PCR), biochemical, haematological and immunological parameters. The control diet fed F. indicus succumbed to death 100% within 3 days whereas the D-3 and D-4 helped to reduced the cumulative mortality of 60-80% respectively. The qRT-PCR revealed that, the WSSV copy number was gradually decreased when increasing concentration of A. grandiflora extract active fraction in the diets. The diets D-3 and D-4 helped to

  20. Role of Litopenaeus vannamei Yin Yang 1 in the Regulation of the White Spot Syndrome Virus Immediate Early Gene ie1.

    PubMed

    Huang, Ping-Han; Huang, Ting-Yi; Cai, Pei-Si; Chang, Li-Kwan

    2017-03-15

    Yin Yang 1 (YY1) is a multifunctional zinc finger transcription factor that regulates many key cellular processes. In this study, we report the cloning of YY1 from Litopenaeus vannamei shrimp (LvYY1). This study shows that LvYY1 is ubiquitously expressed in shrimp tissues, and knockdown of LvYY1 expression by double-stranded RNA (dsRNA) injection in white spot syndrome virus (WSSV)-infected shrimp reduced both mRNA levels of the WSSV immediate early gene ie1 as well as overall copy numbers of the WSSV genome. The cumulative mortality rate of infected shrimp also declined with LvYY1 dsRNA injection. Using an insect cell model, we observed that LvYY1 activates ie1 expression, and a mutation introduced into the ie1 promoter subsequently repressed this capability. Moreover, reporter assay results suggested that LvYY1 is involved in basal transcriptional regulation via an interaction with L. vannamei TATA-binding protein (LvTBP). Electrophoretic mobility shift assay (EMSA) results further indicated that LvYY1 binds to a YY1-binding site in the region between positions -119 and -126 in the ie1 promoter. Chromatin immunoprecipitation analysis also confirmed that LvYY1 binds to the ie1 promoter in WSSV-infected shrimp. Taken together, these results indicate that WSSV uses host LvYY1 to enhance ie1 expression via a YY1-binding site and the TATA box in the ie1 promoter, thereby facilitating lytic activation and viral replication.IMPORTANCE WSSV has long been a scourge of the shrimp industry and remains a serious global threat. Thus, there is a pressing need to understand how the interactions between WSSV and its host drive infection, lytic development, pathogenesis, and mortality. Our successful cloning of L. vannamei YY1 (LvYY1) led to the elucidation of a critical virus-host interaction between LvYY1 and the WSSV immediate early gene ie1 We observed that LvYY1 regulates ie1 expression via a consensus YY1-binding site and TATA box. LvYY1 was also found to interact with L

  1. Mongolian spots.

    PubMed

    Gupta, Divya; Thappa, Devinder Mohan

    2013-01-01

    Mongolian spots (MS) are birthmarks that are present at birth and their most common location is sacrococcygeal or lumbar area. Lesions may be single or multiple and usually involve < 5% total body surface area. They are macular and round, oval or irregular in shape. The color varies from blue to greenish, gray, black or a combination of any of the above. The size varies from few to more than 20 centimetres. Pigmentation is most intense at the age of one year and gradually fades thereafter. It is rarely seen after the age of 6 years. Aberrant MS over occiput, temple, mandibular area, shoulders and limbs may be confused with other dermal melanocytoses and bruises secondary to child abuse, thus necessitating documentation at birth. Although regarded as benign, recent data suggest that MS may be associated with inborn errors of metabolism and neurocristopathies. Mongolian spots usually resolve by early childhood and hence no treatment is generally needed if they are located in the sacral area. However, sometimes it may be required for extrasacral lesions for cosmesis.

  2. Dietary supplementation of probiotic Bacillus PC465 isolated from the gut of Fenneropenaeus chinensis improves the health status and resistance of Litopenaeus vannamei against white spot syndrome virus.

    PubMed

    Chai, Peng-Cheng; Song, Xiao-Ling; Chen, Guo-Fu; Xu, Hua; Huang, Jie

    2016-07-01

    days 15 and 30 post-feeding. In two white spot syndrome virus (WSSV) infections, the weight gain, survival, and WSSV copies within the gills of the probiotic-treated shrimp significantly differed (p < 0.05) from those of the control group. Relatively efficient protection was associated with probiotic feeding. Results suggested that Bacillus PC465 feeding improves the growth performance, survival, digestion, and nutrient absorption of L. vannamei. Probiotic treatment also enhances the microbial structures in the gut, promotes the immune status of shrimp, and provides protection against viral infection. The supplementation with 10(9) CFU g(-1) can also improve the growth and survival of L. vannamei.

  3. Southern Spots

    NASA Technical Reports Server (NTRS)

    2005-01-01

    [figure removed for brevity, see original site] Context image for PIA03092 Southern Spots

    This VIS image of the south polar region was collected during the summer season. The markings of the pole are very diverse and easy to see after the winter frost has been removed.

    Image information: VIS instrument. Latitude 79.7S, Longitude 56.6E. 17 meter/pixel resolution.

    Note: this THEMIS visual image has not been radiometrically nor geometrically calibrated for this preliminary release. An empirical correction has been performed to remove instrumental effects. A linear shift has been applied in the cross-track and down-track direction to approximate spacecraft and planetary motion. Fully calibrated and geometrically projected images will be released through the Planetary Data System in accordance with Project policies at a later time.

    NASA's Jet Propulsion Laboratory manages the 2001 Mars Odyssey mission for NASA's Office of Space Science, Washington, D.C. The Thermal Emission Imaging System (THEMIS) was developed by Arizona State University, Tempe, in collaboration with Raytheon Santa Barbara Remote Sensing. The THEMIS investigation is led by Dr. Philip Christensen at Arizona State University. Lockheed Martin Astronautics, Denver, is the prime contractor for the Odyssey project, and developed and built the orbiter. Mission operations are conducted jointly from Lockheed Martin and from JPL, a division of the California Institute of Technology in Pasadena.

  4. Are 'hot spots' hot spots?

    NASA Astrophysics Data System (ADS)

    Foulger, Gillian R.

    2012-07-01

    The term 'hot spot' emerged in the 1960s from speculations that Hawaii might have its origins in an unusually hot source region in the mantle. It subsequently became widely used to refer to volcanic regions considered to be anomalous in the then-new plate tectonic paradigm. It carried with it the implication that volcanism (a) is emplaced by a single, spatially restricted, mongenetic melt-delivery system, assumed to be a mantle plume, and (b) that the source is unusually hot. This model has tended to be assumed a priori to be correct. Nevertheless, there are many geological ways of testing it, and a great deal of work has recently been done to do so. Two fundamental problems challenge this work. First is the difficulty of deciding a 'normal' mantle temperature against which to compare estimates. This is usually taken to be the source temperature of mid-ocean ridge basalts (MORBs). However, Earth's surface conduction layer is ˜200 km thick, and such a norm is not appropriate if the lavas under investigation formed deeper than the 40-50 km source depth of MORB. Second, methods for estimating temperature suffer from ambiguity of interpretation with composition and partial melt, controversy regarding how they should be applied, lack of repeatability between studies using the same data, and insufficient precision to detect the 200-300 °C temperature variations postulated. Available methods include multiple seismological and petrological approaches, modelling bathymetry and topography, and measuring heat flow. Investigations have been carried out in many areas postulated to represent either (hot) plume heads or (hotter) tails. These include sections of the mid-ocean spreading ridge postulated to include ridge-centred plumes, the North Atlantic Igneous Province, Iceland, Hawaii, oceanic plateaus, and high-standing continental areas such as the Hoggar swell. Most volcanic regions that may reasonably be considered anomalous in the simple plate-tectonic paradigm have been

  5. Mutational Hot Spot Potential of a Novel Base Pair Mutation of the CSPG2 Gene in a Family With Wagner Syndrome

    PubMed Central

    Ronan, Shawn M.; Tran-Viet, Khanh-Nhat; Burner, Erica L.; Metlapally, Ravikanth; Toth, Cynthia A.; Young, Terri L.

    2012-01-01

    Objective To report a 3-generation white family clinically diagnosed variably with Wagner, Stickler, and Jansen syndromes and screened for sequence variants in the COL2A1 and CSPG2 genes. Wagner syndrome is an autosomal dominant vitreoretinopathy with a predisposition to retinal detachment and cataracts. It has significant phenotypic overlap with allelic Jansen syndrome and ocular Stickler syndrome type 1. Sticker syndrome type 1 maps to chromosome 12q13.11-q13.2, with associated COL2A1 gene mutations. Wagner syndrome maps to chromosome 5q13-q14 and is associated with mutations in CSPG2 encoding versican, a proteoglycan present in human vitreous. Methods Genomic DNA samples derived from venous blood were collected from all family members. Complete sequencing of COL2A1 was performed on a proband. Primers for polymerase chain reaction and sequencing were designed to cover all exon and intronexon boundaries. Direct sequencing of CSPG2 was performed on all family member samples. Results No detectable COL2A1 mutations were noted, making the diagnosis of ocular Stickler syndrome highly unlikely for this family. A unique base pair substitution (c.9265+1G>T) in intron 8 of the CSPG2 gene cosegregating with disease status was identified. This mutation occurred in a highly conserved previously reported splice site with a similar base pair substitution(G>A). Direct sequencing of this splice site mutation in 107 unrelated external controls revealed no variants, supporting the rarity of this base pair change and its causation in Wagner syndrome. This novel base pair substitution is thought to cause the deletion of exon 8 and formation of a truncated protein product. Conclusion Mutation screening of CSPG2 in autosomal dominant vitreoretinopathy families is important for accurate diagnosis. Clinical Relevance This study underscores the importance of obtaining extensive pedigree information and comparative ophthalmologic clinical information, as the phenotypic findings may vary

  6. Mongolian blue spots (image)

    MedlinePlus

    Mongolian blue spots are flat bluish- to bluish-gray skin markings commonly appearing at birth or shortly ... back and also can appear on the shoulders. Mongolian spots are benign and are not associated with ...

  7. Rocky Mountain spotted fever

    MedlinePlus

    ... this page: //medlineplus.gov/ency/article/000654.htm Rocky Mountain spotted fever To use the sharing features on this page, please enable JavaScript. Rocky Mountain spotted fever is a disease caused by a type of ...

  8. Interaction between shrimp and white spot syndrome virus through PmRab7-VP28 complex: an insight using simulation and docking studies.

    PubMed

    Verma, Arunima Kumar; Gupta, Shipra; Verma, Sharad; Mishra, Abha; Nagpure, N S; Singh, Shivesh Pratap; Pathak, Ajey Kumar; Sarkar, Uttam Kumar; Singh, Shri Prakash; Singh, Mahender; Seth, Prahlad Kishore

    2013-03-01

    White spot disease is a devastating disease of shrimp Penaeus monodon in which the shrimp receptor protein PmRab7 interacts with viral envelop protein VP28 to form PmRab7-VP28 complex, which causes initiation of the disease. The molecular mechanism implicated in the disease, the dynamic behavior of proteins as well as interaction between both the biological counterparts that crafts a micro-environment feasible for entry of virus into the shrimp is still unknown. In the present study, we applied molecular modeling (MM), molecular dynamics (MD) and docking to compute surface mapping of infective amino acid residues between interacting proteins. Our result showed that α-helix of PmRab7 (encompassing Ser74, Ile143, Thr184, Arg53, Asn144, Thr184, Arg53, Arg79) interacts with β-sheets of VP28 (containing Ser74, Ile143, Thr184, Arg53, Asn144, Thr184, Arg53, Arg79) and Arg69-Ser74, Val75-Ile143, Leu73-Ile143, Arg79-Asn144, Ala198-Ala182 bonds contributed in the formation of PmRab7-VP28 complex. Further studies on the amino acid residues and bonds may open new possibilities for preventing PmRab7-VP28 complex formation, thus reducing chances of WSD. The quantitative predictions provide a scope for experimental testing in future as well as endow with a straightforward evidence to comprehend cellular mechanisms underlying the disease.

  9. Multiplex real-time PCR and high-resolution melting analysis for detection of white spot syndrome virus, yellow-head virus, and Penaeus monodon densovirus in penaeid shrimp.

    PubMed

    Panichareon, Benjaporn; Khawsak, Paisarn; Deesukon, Warin; Sukhumsirichart, Wasana

    2011-12-01

    A multiplex real-time PCR and high-resolution melting (HRM) analysis was developed to detect simultaneously three of the major viruses of penaeid shrimp including white spot syndrome virus (WSSV), yellow-head virus (YHV), and Penaeus monodon densovirus (PmDNV). Plasmids containing DNA/cDNA fragments of WSSV and YHV, and genomic DNAs of PmDNV and normal shrimp were used to test sensitivity of the procedure. Without the need of any probe, the products were identified by HRM analysis after real-time PCR amplification using three sets of viral specific primers. The results showed DNA melting curves that were specific for individual virus. No positive result was detected with nucleic acids from shrimp, Penaeus monodon nucleopolyhedrovirus (PemoNPV), Penaeus stylirostris densovirus (PstDNV), or Taura syndrome virus (TSV). The detection limit for PmDNV, YHV and WSSV DNAs were 40fg, 50fg, and 500fg, respectively, which was 10 times more sensitive than multiplex real-time PCR analyzed by agarose gel electrophoresis. In viral nucleic acid mixtures, HRM analysis clearly identified each virus in dual and triple infection. To test the capability to use this method in field, forty-one of field samples were examined by HRM analysis in comparison with agarose gel electrophoresis. For HRM analysis, 11 (26.83%), 9 (21.95%), and 4 (9.76%) were infected with WSSV, PmDNV, and YHV, respectively. Agarose gel electrophoresis detected lesser number of PmDNV infection which may due to the limit of sensitivity. No multiple infection was found in these samples. This method provides a rapid, sensitive, specific, and simultaneous detection of three major viruses making it as a useful tool for diagnosis and epidemiological studies of these viruses in shrimp and carriers.

  10. Development of a rapid method for identifying carryover contamination of positive control DNA, using a chimeric positive control and restriction enzyme for the diagnosis of white spot syndrome virus by nested PCR.

    PubMed

    Kim, Hyoung Jun; Kwon, Se Ryun

    2014-12-01

    Chimeric positive plasmids have been developed to minimize false-positive reactions caused by polymerase chain reaction (PCR) contamination. Here, we developed a rapid method for identifying false-positive results while detecting white spot syndrome virus (WSSV) by nested PCR, using chimeric positive plasmids. The results of PCRs using WSSV diagnostic primer sets showed PCR products of a similar size (WSSV 1st PCR product, 1,447 bp; WSSV 2nd PCR product, 941 bp) using WSSV chimeric plasmids or DNA from shrimp infected with WSSV. The PCR products were digested with DraI for 1 h at 37 °C. The digested chimeric DNA separated into two DNA bands; however, the WSSV-infected shrimp DNA did not separate. Thus, chimeric plasmid DNA may be used as positive control DNA instead of DNA from WSSV-infected shrimp, in order to prevent PCR contamination. Thus, the use of restriction enzyme digestion allowed us to rapidly distinguish between WSSV DNA and WSSV chimeric plasmid DNA.

  11. Astakine LvAST binds to the β subunit of F1-ATP synthase and likely plays a role in white shrimp Litopeneaus vannamei defense against white spot syndrome virus.

    PubMed

    Liang, Gao-Feng; Liang, Yan; Xue, Qinggang; Lu, Jin-Feng; Cheng, Jun-Jun; Huang, Jie

    2015-03-01

    Cytokines play a critical role in innate and adaptive immunity. Astakines represent a group of invertebrate cytokines that are related to vertebrate prokineticin and function in promoting hematopoiesis in crustaceans. We have identified an astakine from the white shrimp Litopeneaus vannamei and named it LvAST in a previous research. In the present research, we investigated the interactions among LvAST, the envelope protein VP37 of white spot syndrome virus (i.e., WSSV), and the β subunit of F1-ATP synthase (ATPsyn-β) of the white shrimp (i.e., BP53) using binding assays and co-precipitations. We also examined the effects of LvAST on shrimp susceptibility to WSSV. We found that LvAST and VP37 competitively bound to BP53, but did not bind to each other. Shrimps that had been injected with recombinant LvAST exhibited significantly lower mortality and longer survival time in experimental infections by WSSV. In contrast, shrimps whose LvAST gene expression had been inhibited by RNA interference showed significantly higher WSSV infection intensity and shorter survival time following viral challenges. These results suggested that LvAST and WSSV both likely use ATPsyn-β as a receptor and LvAST plays a role in shrimp defense against WSSV infection. This represented the first research showing the involvement of astakines in host antiviral immunity.

  12. The Earth's Hot Spots.

    ERIC Educational Resources Information Center

    Vink, Gregory E.; And Others

    1985-01-01

    Hot spots are isolated areas of geologic activity where volcanic eruptions, earthquakes, and upwelling currents occur far from plate boundaries. These mantle plumes are relatively stable and crustal plates drift over them. The nature and location of hot spots (with particular attention to the Hawaiian Islands and Iceland) are discussed. (DH)

  13. Watermarking spot colors

    NASA Astrophysics Data System (ADS)

    Alattar, Osama M.; Reed, Alastair M.

    2003-06-01

    Watermarking of printed materials has usually focused on process inks of cyan, magenta, yellow and black (CMYK). In packaging, almost three out of four printed materials include spot colors. Spot colors are special premixed inks, which can be produced in a vibrant range of colors, often outside the CMYK color gamut. In embedding a watermark into printed material, a common approach is to modify the luminance value of each pixel in the image. In the case of process color work pieces, the luminance change can be scaled to the C, M, Y and K channels using a weighting function, to produce the desired change in luminance. In the case of spot color art designs, there is only one channel available and the luminance change is applied to this channel. In this paper we develop a weighting function to embed the watermark signal across the range of different spot colors. This weighting function normalizes visibility effect and signal robustness across a wide range of different spot colors. It normalizes the signal robustness level over the range of an individual spot color"s intensity levels. Further, it takes into account the sensitivity of the capturing device to the different spot colors.

  14. Neuroendocrine responses of a crustacean host to viral infection: effects of infection of white spot syndrome virus on the expression and release of crustacean hyperglycemic hormone in the crayfish Procambarus clarkii.

    PubMed

    Lin, Ling-Jiun; Chen, Yan-Jhou; Chang, Yun-Shiang; Lee, Chi-Ying

    2013-02-01

    The objectives of the present study were to characterize the changes in crustacean hyperglycemic hormone (CHH) transcript and peptide levels in response to infection of white spot syndrome virus (WSSV) in a crustacean, Procambarus clarkii. After viral challenge, significant increase in virus load began at 24 h post injection (hpi) and the increase was much more substantial at 48 and 72 hpi. The hemolymph CHH levels rapidly increased after viral challenge; the increase started as early as 3 hpi and lasted for at least 2 d after the challenge. In contrast, the hemolymph glucose levels did not significantly changed over a 2 d period in the WSSV-infected animals. The CHH transcript and peptide levels in tissues were also determined. The CHH transcript levels in the eyestalk ganglia (the major site of CHH synthesis) of the virus-infected animals did not significantly change over a 2 d period and those in 2 extra-eyestalk tissues (the thoracic ganglia and cerebral ganglia) significantly increased at 24 and 48 hpi. The CHH peptide levels in the eyestalk ganglia of the virus-infected animals significantly decreased at 24 and 48 hpi and those in the thoracic ganglia and cerebral ganglia remained unchanged over a 2 d period. These data demonstrated a WSSV-induced increase in the release of CHH into hemolymph that is rapid in onset and lasting in duration. Changes in the CHH transcript and peptide levels implied that the WSSV-induced increase in hemolymph CHH levels primarily resulted from an enhanced release from the eyestalk ganglia, but the contribution of the 2 extra-eyestalk tissues to hemolymph pool of CHH increased as viral infection progressed. The combined patterns of change in the hemolymph glucose and CHH levels further suggest that the virus-enhanced CHH release would lead to higher glycolytic activity and elevated glucose mobilization presumably favorable for viral replication.

  15. Mononucleosis spot test

    MedlinePlus

    Monospot test; Heterophile antibody test; Heterophile agglutination test; Paul-Bunnell test; Forssman antibody test ... The mononucleosis spot test is done when symptoms of mononucleosis are ... Fatigue Fever Large spleen (possibly) Sore throat Tender ...

  16. Lincoln's Spot Resolutions.

    ERIC Educational Resources Information Center

    Mueller, Jean West; Schamel, Wynell Burroughs

    1988-01-01

    Examines the events leading to and immediately following the declaration of war on Mexico in 1846. Includes the second and third pages of Abraham Lincoln's "Spot Resolutions" and presents teaching suggestions for interpreting the document and assessing public opinion. (GEA)

  17. SPOT4 Management Centre

    NASA Technical Reports Server (NTRS)

    Labrune, Yves; Labbe, X.; Roussel, A.; Vielcanet, P.

    1994-01-01

    In the context of the CNES SPOT4 program CISI is particularly responsible for the development of the SPOT4 Management Centre, part of the SPOT4 ground control system located at CNES Toulouse (France) designed to provide simultaneous control over two satellites. The main operational activities are timed to synchronize with satellite visibilities (ten usable passes per day). The automatic capability of this system is achieved through agenda services (sequence of operations as defined and planned by operator). Therefore, the SPOT4 Management Centre offers limited, efficient and secure human interventions for supervision and decision making. This paper emphasizes the main system characteristics as degree of automation, level of dependability and system parameterization.

  18. Scientists Spot 'Teetotaler' Gene

    MedlinePlus

    ... gov/news/fullstory_162265.html Scientists Spot 'Teetotaler' Gene Discovery might one day lead to drugs to ... HealthDay News) -- Scientists say they've identified a gene variant that dampens the desire to drink alcohol. ...

  19. Dietary administration of Gynura bicolor (Roxb. Willd.) DC water extract enhances immune response and survival rate against Vibrio alginolyticus and white spot syndrome virus in white shrimp Litopeneaus vannamei.

    PubMed

    Wu, Chih-Chung; Chang, Yueh-Ping; Wang, Jyh-Jye; Liu, Chun-Hung; Wong, Saou-Lien; Jiang, Chii-Ming; Hsieh, Shu-Ling

    2015-01-01

    Gynura bicolor (Roxb. & Willd.) DC., a perennial plant belonging to the Asteraceae family, is originated from the tropical area of Asia. The total hemocyte count (THC), phenoloxidase (PO) activity, respiratory bursts (RBs), superoxide dismutase (SOD) activity, and lysozyme activity were examined after white shrimp Litopenaeus vannamei had been fed diets containing the water extract of G. bicolor at 0 (control), 0.5, 1.0, and 2.0 g (kg diet)(-1) for 7-28 days. The results indicated that these parameters increased accordingly with the amount of extract and time. THCs of the shrimp fed the G. bicolor diets at 1.0 and 2.0 g (kg diet)(-1) were significantly higher than that fed the control diet for 14-28 days. For the shrimp fed the G. bicolor diets at 0.5, 1.0, and 2.0 g (kg diet)(-1), the PO, RBs, and lysozyme activities reached the highest levels after 7 days, whereas SOD activity reached the highest levels after 14 days. In a separate experiment, white shrimp L. vannamei fed the diets containing the G. bicolor extract for 28 days were challenged with Vibrio alginolyticus at 3 × 10(6) cfu shrimp(-1) and white spot syndrome virus (WSSV) at 1 × 10(3) copies shrimp(-1). The survival rate of the shrimp fed the G. bicolor diets was significantly higher than that of the shrimp fed the control diet at 48-144 h post challenge V. alginolyticus and WSSV. For the shrimp fed the G. bicolor diets at 0.5, 1 and 2 g (kg diet)(-1) under challenges of V. alginolyticus and WSSV, their LPS- and β-1,3-glucan-binding protein (LGBP) and peroxinectin (PE) mRNA expressions were significantly higher than those of the challenged control shrimp at 12-96 and 24-144 h post-challenge, respectively. We concluded that dietary administration of a G. bicolor extract could enhance the innate immunity within 28 days as evidenced by the increases in immune parameters (PO, RBs, and lysozyme) and antioxidant enzyme (SOD) activities of shrimp to against V. alginolyticus and WSSV

  20. IR Spot Weld Inspect

    SciTech Connect

    Chen, Jian; Feng, Zhili

    2014-01-01

    In automotive industry, destructive inspection of spot welds is still the mandatory quality assurance method due to the lack of efficient non-destructive evaluation (NDE) tools. However, it is costly and time-consuming. Recently at ORNL, a new NDE prototype system for spot weld inspection using infrared (IR) thermography has been developed to address this problem. This software contains all the key functions that ensure the NDE system to work properly: system input/output control, image acquisition, data analysis, weld quality database generation and weld quality prediction, etc.

  1. Rolling Spot Welder

    NASA Technical Reports Server (NTRS)

    Wagner, Garret E.; Fonteyne, Steve L.

    1990-01-01

    Wheeled tool speeds tack-welding operations. Spotwelds foil to parts in preparation for brazing. Includes electrode wheel rolling across foil. Welding current in electrode pulsed as electrode moves along, making series of uniformly-spaced low-current spot welds.

  2. Bacterial leaf spot

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacterial leaf spot has been reported in Australia (Queensland), Egypt, El Salvador, India, Japan, Nicaragua, Sudan, and the United States (Florida, Iowa, Kansas, Maryland, and Wisconsin). It occasionally causes locally severe defoliation and post-emergence damping-off and stunting. The disease is...

  3. Rocky Mountain Spotted Fever.

    PubMed

    Phillips, Jennan

    2017-01-01

    The tick-borne disease Rocky Mountain spotted fever (RMSF) can have deadly outcomes unless treated appropriately, yet nonspecific flu-like symptoms complicate diagnosis. Occupational health nurses must have a high index of suspicion with symptomatic workers and recognize that recent recreational or occupational activities with potential tick exposure may suggest RMSF.

  4. Arc spot grouping: An entanglement of arc spot cells

    SciTech Connect

    Kajita, Shin; Hwangbo, Dogyun; Ohno, Noriyasu; Tsventoukh, Mikhail M.; Barengolts, Sergey A.

    2014-12-21

    In recent experiments, clear transitions in velocity and trail width of an arc spot initiated on nanostructured tungsten were observed on the boundary of the thick and thin nanostructured layer regions. The velocity of arc spot was significantly decreased on the thick nanostructured region. It was suggested that the grouping decreased the velocity of arc spot. In this study, we try to explain the phenomena using a simple random walk model that has properties of directionality and self-avoidance. And grouping feature was added by installing an attractive force between spot cells with dealing with multi-spots. It was revealed that an entanglement of arc spot cells decreased the spot velocity, and spot cells tend to stamp at the same location many times.

  5. TV spots' impact.

    PubMed

    El-bakly, S

    1994-09-01

    The Information, Education and Communication (IEC) Center of the State Information Service was established in 1979 for the purpose of providing information to the people on the population issue. The Ministry of Information has accorded the State Information Service free TV and radio air time for family planning dramas and spots. In the early years information campaigns were organized to make people aware of the population problem by slogans, songs, and cartoons. Around 1984 misconceptions about family planning and contraceptives were attacked through a number of TV and radio spots. A few years later 21 spots on specific contraceptive methods were broadcast which were aired for three years over 3000 times. They were extremely successful. The impact of these TV spots was one of the major reasons why the contraceptive prevalence rate increased from 30% in 1984 to 38% in 1988 and 47% in 1992. Spots were also broadcast about the social implications of large families. The TV soap opera "And The Nile Flows On", with the family planning message interwoven into it, was very well received by the target audience. A program entitled "Wedding of the Month" features couples who know family planning well. The most successful radio program is a 15-20 minute long quiz show for residents of the villages where the Select Villages Project is being implemented. The State Information Service has 60 local information centers in the 26 governorates of Egypt that make plans for the family planning campaign. In 1992 the Minya Initiative, a family planning project was implemented in the Minya Governorate. As a result, the contraceptive prevalence rate rose from 22% to 30% over 18 months. A new project, the Select Village Project, was developed in 1993 that replicates the Minya Initiative on the village level in other governorates. This new project that was implemented in sixteen governorates.

  6. Human Infection with Novel Spotted Fever Group Rickettsia Genotype, China, 2015

    PubMed Central

    Li, Hao; Cui, Xiao-Ming; Cui, Ning; Yang, Zhen-Dong; Hu, Jian-Gong; Fan, Ya-Di; Fan, Xue-Juan; Zhang, Lan; Zhang, Pan-He

    2016-01-01

    Only 4 species of spotted fever group rickettsiae have been detected in humans in China. However, phylogenetic analysis of samples from 5 ill patients in China indicated infection with a novel spotted fever group Rickettsia, designated Rickettsia sp. XY99. Clinical signs resembled those of severe fever with thrombocytopenia syndrome. PMID:27869588

  7. Poisson Spot with Magnetic Levitation

    ERIC Educational Resources Information Center

    Hoover, Matthew; Everhart, Michael; D'Arruda, Jose

    2010-01-01

    In this paper we describe a unique method for obtaining the famous Poisson spot without adding obstacles to the light path, which could interfere with the effect. A Poisson spot is the interference effect from parallel rays of light diffracting around a solid spherical object, creating a bright spot in the center of the shadow.

  8. Rocky Mountain spotted fever.

    PubMed

    Lacz, N L; Schwartz, R A; Kapila, R

    2006-04-01

    Rocky Mountain spotted fever (RMSF) is an unusual but important dermatological condition to identify without hesitation. The classic triad of headache, fever, and a rash that begins on the extremities and travels proximally to involve the trunk is found in a majority of patients. The cutaneous centripetal pattern is a result of cell to cell migration by the causative organism Rickettsia rickettsii. Such individuals should receive prompt antimicrobial therapy and supportive care to avoid serious and potentially fatal complications.

  9. Rocky Mountain spotted fever.

    PubMed

    Kamper, C A; Chessman, K H; Phelps, S J

    1988-02-01

    The epidemiology, pathogenesis, clinical features, and treatment of Rocky Mountain spotted fever are reviewed. Rocky Mountain spotted fever is a severe infection caused by Rickettsia rickettsii transmitted to man by various species of ticks. High-incidence areas exist in the southeast and south central United States. Only 60-70% of patients with the disease report a history of tick bite or exposure to tick-infested areas. The disease is initially characterized by fever, headache, gastrointestinal complaints, myalgia, and a generalized rash. In several days generalized vasculitis may lead to periorbital edema and nonpitting edema of the face and extremities. Central nervous system involvement is common. Because signs and symptoms associated with the disease are nonspecific, the diagnosis is often delayed or missed. Traditionally diagnostic confirmation relied on serologic testing, but an indirect fluorescent antibody assay will soon be commercially available. Rocky Mountain spotted fever is usually treated with the rickettsiostatic agents chloramphenicol or tetracycline, but few comparative data on these agents in patients with the disease are available. For patients who cannot tolerate oral medications, intravenous chloramphenicol sodium succinate is the preferred treatment; chloramphenicol is also the drug of choice for children less than eight years of age. Otherwise, oral tetracycline hydrochloride is the drug of choice. Antibiotic therapy should be continued for 7-10 days or until the patient is afebrile for two to five days. All cases of Rocky Mountain spotted fever must be reported to the Centers for Disease Control. The best ways to decrease the morbidity and mortality of the disease are to increase awareness of its signs and symptoms and to prevent exposure to ticks.

  10. Mongolian spots: How important are they?

    PubMed

    Gupta, Divya; Thappa, Devinder Mohan

    2013-11-16

    Mongolian spots (MS) are congenital birthmarks seen most commonly over the lumbosacral area. They are bluish-green to black in color and oval to irregular in shape. They are most commonly found in individuals of African or Asian ethnic background. Although these lesions resolve by one to two years of age, widespread, extrasacral and dark colored MS sometimes persist into adulthood. Aberrant MS over occiput, temple, mandibular area, shoulders and limbs may be confused with other dermal melanocytoses and bruises secondary to child abuse, thus necessitating documentation at birth. Although traditionally believed to be benign in nature, they have now been shown to co-exist with inborn errors of metabolism, most commonly GM1 gangliosidosis and mucopolysaccharidosis type I (Hurler's disease), followed by mucopolysaccharidosis type II (Hunter's syndrome), mucolipidosis, Niemann-Pick disease and mannosidosis. They have also been seen to co-exist with various vascular or other pigmented birthmarks like café-au-lait macules. Co-existing Mongolian spots and vascular birthmarks like nevus flammeus, nevus anemicus or nevus spilus is termed as phakomatosis pigmentovascularis. This review focuses on the important associations of Mongolian spots and stresses upon the importance of screening babies with extensive MS.

  11. Red Spot Movie

    NASA Technical Reports Server (NTRS)

    2000-01-01

    This brief movie shows counterclockwise atmospheric motion around Jupiter's Great Red Spot. The clip was made from blue-filter images taken with the narrow-angle camera on NASA's Cassini spacecraft during seven separate rotations of Jupiter between Oct. 1 and Oct. 5, 2000.

    The clip also shows the eastward and westward motion of the zonal jets, seen as the horizontal stripes flowing in opposite directions. The zonal jets circle the planet. As far as can be determined from both Earth-based and spacecraft measurements, the positions and speeds of the jets have not changed for 100 years. Since Jupiter is a fluid planet without a solid boundary, the jet speeds are measured relative to Jupiter's magnetic field, which rotates, wobbling like a top because of its tilt, every 9 hours 55.5 minutes. The movie shows motions in the magnetic reference frame, so winds to the west correspond to features that are rotating a little slower than the magnetic field, and eastward winds correspond to features rotating a little faster.

    Because the Red Spot is in the southern hemisphere, the direction of motion indicates it is a high-pressure center. Small bright clouds appear suddenly to the west of the Great Red Spot. Scientists suspect these small white features are lightning storms. The storms eventually merge with the Red Spot and surrounding jets, and may be the main energy source for the large-scale features.

    The smallest features in the movie are about 500 kilometers (about 300 miles) across. The spacing of the movie frames in time is not uniform; some consecutive images are separated by two Jupiter rotations, and some by one. The images have been re-projected using a simple cylindrical map projection. They show an area from 50 degrees north of Jupiter's equator to 50 degrees south, extending 100 degrees east-west, about one quarter of Jupiter's circumference.

    Cassini is a cooperative project of NASA, the European Space Agency and the Italian Space Agency. The Jet

  12. [Rocky Mountain spotted fever].

    PubMed

    Reinauer, K M; Jaschonek, K; Kusch, G; Heizmann, W R; Döller, P C; Jenss, H

    1990-01-12

    After returning from a holiday in the USA a 24-year-old man fell ill with diarrhoea, high fever and marked rash including the palms of the hands and soles of the feet. When a history of a tick bite in the USA was elicited, a rickettsial infection was suspected. Treatment with doxycycline, 100 mg twice daily, was instituted finally and the fever slowly resolved. The patient became completely well again within four weeks. Serological tests confirmed the diagnosis of Rocky Mountain spotted fever.

  13. Dark Spots and Fans

    NASA Technical Reports Server (NTRS)

    2006-01-01

    As winter turns to spring at the south polar ice cap of Mars, the rising sun reveals dark spots and fans emerging from the cold polar night. Using visual images (left) and temperature data (right) from the Thermal Emission Imaging system on NASA's Mars Odyssey orbiter, scientists have built a new model for the origin of the dark markings. Scientists propose the markings come from dark sand and dust strewn by high-speed jets of carbon-dioxide gas. These erupt from under a layer of carbon-dioxide ice that forms each Martian winter.

  14. Configurable hot spot fixing system

    NASA Astrophysics Data System (ADS)

    Kajiwara, Masanari; Kobayashi, Sachiko; Mashita, Hiromitsu; Aburada, Ryota; Furuta, Nozomu; Kotani, Toshiya

    2014-03-01

    Hot spot fixing (HSF) method has been used to fix many hot spots automatically. However, conventional HSF based on a biasing based modification is difficult to fix many hot spots under a low-k1 lithography condition. In this paper we proposed a new HSF, called configurable hotspot fixing system. The HSF has two major concepts. One is a new function to utilize vacant space around a hot spot by adding new patterns or extending line end edges around the hot spot. The other is to evaluate many candidates at a time generated by the new functions. We confirmed the proposed HSF improves 73% on the number of fixing hot spots and reduces total fixing time by 50% on a device layout equivalent to 28nm-node. The result shows the proposed HSF is effective for layouts under the low-k1 lithography condition.

  15. Turbulent spots in hypervelocity flow

    NASA Astrophysics Data System (ADS)

    Jewell, Joseph S.; Leyva, Ivett A.; Shepherd, Joseph E.

    2017-04-01

    The turbulent spot propagation process in boundary layer flows of air, nitrogen, carbon dioxide, and air/carbon dioxide mixtures in thermochemical nonequilibrium at high enthalpy is investigated. Experiments are performed in a hypervelocity reflected shock tunnel with a 5-degree half-angle axisymmetric cone instrumented with flush-mounted fast-response coaxial thermocouples. Time-resolved and spatially demarcated heat transfer traces are used to track the propagation of turbulent bursts within the mean flow, and convection rates at approximately 91, 74, and 63% of the boundary layer edge velocity, respectively, are observed for the leading edge, peak, and trailing edge of the spots. A simple model constructed with these spot propagation parameters is used to infer spot generation rates from observed transition onset to completion distance. Spot generation rates in air and nitrogen are estimated to be approximately twice the spot generation rates in air/carbon dioxide mixtures.

  16. Jovian Dark Spot

    NASA Technical Reports Server (NTRS)

    1998-01-01

    A recently discovered black spot in Jupiter's clouds is darker than any feature ever before observed on the giant planet. The spot may be the result of a downward spiraling wind that blows away high clouds and reveals deeper, very dark cloud layers. These three panels depict the same area of Jupiter's atmosphere. A map of Jovian temperatures near 250 millibar pressure (top) panel is derived from the photopolarimeter-radiometer instrument on NASA's Galileo Jupiter orbiter. This map is compared with maps derived from images of the same area in visible light (middle panel)and thermal radiation sensitive to cloud-top temperatures (bottom panel).

    The single downward-pointing arrow in the top panel indicates the location of a warm area that corresponds to the position of a so-called 'black spot'(shown in the middle panel), a feature that is about a year old. Features this dark are rare on Jupiter. The bottom panel, sensitive to temperatures at Jupiter's cloud tops, shows this feature as a bright object, meaning that upper-level cold clouds are missing - allowing us to see deeper into Jupiter's warmer interior. The dark visible appearance of the feature than most likely represents the color of very deep clouds. The warm temperatures and cloud-free conditions imply that this feature is a region where dry upper-atmospheric gas is being forced to converge, is warmed up and then forced to descend, clearing out clouds. It is the opposite of wet, upwelling gas in areas such as Jupiter's Great Red Spot or white ovals. On the other hand, it is unlike the dry and relatively cloudless feature into which the Galileo probe descended in 1995, because that region had the same temperatures as its surroundings and did not appear nearly as dark as this new spot.

    The temperatures sampled by the photopolarimeter radiometer are near the top of Jupiter's troposphere, where wind motions control the atmosphere. The top row of arrows shows the location of temperature waves in a warm region

  17. Resolving stellar surface spots

    NASA Astrophysics Data System (ADS)

    Strassmeier, K. G.; Carroll, T.; Rice, J. B.; Savanov, I. S.

    Doppler imaging of stellar surfaces is a novel technique with similarities to medical brain tomography (instead of a fixed brain and a rotating scanner, astronomers have a fixed spectrograph and a rotating brain, star of course). The number of free (internal) parameters is of the order of the number of surface grid points and only constrained by the number of input data points. This obviously ill-posed situation requires modern inversion algorithms with penalty functions of the form of maximum entropy or Tikhonov etc.. We present a brief status review of our Doppler imaging codes at AIP that span from temperature and spot-filling-factor mapping to full Stokes-based magnetic field mapping.

  18. Rocky Mountain spotted fever.

    PubMed

    Dantas-Torres, Filipe

    2007-11-01

    Rocky Mountain spotted fever (RMSF) is a life-threatening disease caused by Rickettsia rickettsii, an obligately intracellular bacterium that is spread to human beings by ticks. More than a century after its first clinical description, this disease is still among the most virulent human infections identified, being potentially fatal even in previously healthy young people. The diagnosis of RMSF is based on the patient's history and a physical examination, and often presents a dilemma for clinicians because of the non-specific presentation of the disease in its early course. Early empirical treatment is essential to prevent severe complications or a fatal outcome, and treatment should be initiated even in unconfirmed cases. Because there is no vaccine available against RMSF, avoidance of tick-infested areas is still the best way to prevent the infection.

  19. Rocky Mountain spotted fever, Colombia.

    PubMed

    Hidalgo, Marylin; Orejuela, Leonora; Fuya, Patricia; Carrillo, Pilar; Hernandez, Jorge; Parra, Edgar; Keng, Colette; Small, Melissa; Olano, Juan P; Bouyer, Donald; Castaneda, Elizabeth; Walker, David; Valbuena, Gustavo

    2007-07-01

    We investigated 2 fatal cases of Rocky Mountain spotted fever that occurred in 2003 and 2004 near the same locality in Colombia where the disease was first reported in the 1930s. A retrospective serosurvey of febrile patients showed that > 21% of the serum samples had antibodies aaainst spotted fever group rickettsiae.

  20. Hot Spot Cosmic Accelerators

    NASA Astrophysics Data System (ADS)

    2002-11-01

    length of more than 3 million light-years, or no less than one-and-a-half times the distance from the Milky Way to the Andromeda galaxy, this structure is indeed gigantic. The region where the jets collide with the intergalactic medium are known as " hot spots ". Superposing the intensity contours of the radio emission from the southern "hot spot" on a near-infrared J-band (wavelength 1.25 µm) VLT ISAAC image ("b") shows three distinct emitting areas; they are even better visible on the I-band (0.9 µm) FORS1 image ("c"). This emission is obviously associated with the shock front visible on the radio image. This is one of the first times it has been possible to obtain an optical/near-IR image of synchrotron emission from such an intergalactic shock and, thanks to the sensitivity and image sharpness of the VLT, the most detailed view of its kind so far . The central area (with the strongest emission) is where the plasma jet from the galaxy centre hits the intergalactic medium. The light from the two other "knots", some 10 - 15,000 light-years away from the central "hot spot", is also interpreted as synchrotron emission. However, in view of the large distance, the astronomers are convinced that it must be caused by electrons accelerated in secondary processes at those sites . The new images thus confirm that electrons are being continuously accelerated in these "knots" - hence called "cosmic accelerators" - far from the galaxy and the main jets, and in nearly empty space. The exact physical circumstances of this effect are not well known and will be the subject of further investigations. The present VLT-images of the "hot spots" near 3C 445 may not have the same public appeal as some of those beautiful images that have been produced by the same instruments during the past years. But they are not less valuable - their unusual importance is of a different kind, as they now herald the advent of fundamentally new insights into the mysteries of this class of remote and active

  1. Saturn's Hot Spot

    NASA Technical Reports Server (NTRS)

    2005-01-01

    This is the sharpest image of Saturn's temperature emissions taken from the ground; it is a mosaic of 35 individual exposures made at the W.M. Keck I Observatory, Mauna Kea, Hawaii on Feb. 4, 2004.

    The images to create this mosaic were taken with infrared radiation. The mosaic was taken at a wavelength near 17.65 microns and is sensitive to temperatures in Saturn's upper troposphere. The prominent hot spot at the bottom of the image is right at Saturn's south pole. The warming of the southern hemisphere was expected, as Saturn was just past southern summer solstice, but the abrupt changes in temperature with latitude were not expected. The tropospheric temperature increases toward the pole abruptly near 70 degrees latitude from 88 to 89 Kelvin (-301 to -299 degrees Fahrenheit) and then to 91 Kelvin (-296 degrees Fahrenheit) right at the pole.

    Ring particles are not at a uniform temperature everywhere in their orbit around Saturn. The ring particles are orbiting clockwise in this image. Particles are coldest just after having cooled down in Saturn's shadow (lower left). As they orbit Saturn, the particles increase in temperature up to a maximum (lower right) just before passing behind Saturn again in shadow.

    A small section of the ring image is missing because of incomplete mosaic coverage during the observing sequence.

  2. Single spots, unipolar magnetic regions, and pairs of spots

    NASA Astrophysics Data System (ADS)

    Akasofu, S.-I.

    2014-06-01

    McIntosh (1981) noted that sunspot pairs appear preferentially near the boundary of unipolar magnetic (UM) regions of opposite polarity. A large number of solar magnetograms from the Mount Wilson Observatory and the Kitt Peak Observatory during fairly quiet periods are examined to confirm his finding. In this study, it is also found collaterally that positive single spots appear in a positive UM region and vice versa. It is suggested thus that a pair of spots of opposite polarity is formed because two single spots develop in the vicinity of the boundary (the neutral line) of two UM regions of opposite polarity for polarity arrangement appropriate to the Hale law, namely, the Hale boundary. For these reasons, it is suggested that single spots and UM regions have significant meaning in solar magnetism.

  3. Weird Warm Spot on Exoplanet

    NASA Video Gallery

    This animation illustrates an unexpected warm spot on the surface of a gaseous exoplanet. NASA's Spitzer Space Telescope discovered that the hottest part of the planet, shown here as bright, orange...

  4. Center Spot: Shoe Box Science

    ERIC Educational Resources Information Center

    Hoffman, Jan

    1976-01-01

    This is the second "Center Spot" devoted to Jan Hoffman's "Shoe Box Science," a program that organizes manipulative materials so that children can identify, describe, order, construct, name and distinguish on their own.

  5. Neurofibromatosis type 1 with overlap Turner syndrome and Klinefelter syndrome.

    PubMed

    Hatipoglu, Nihal; Kurtoglu, Selim; Kendirci, Mustafa; Keskin, Mehmet; Per, Hüseyin

    2010-02-01

    Turner's syndrome is a sex chromosome disorder. Klinefelter's syndrome is one of the most severe genetic diseases. Neurofibromatosis is an autosomal dominant disorder characterized by cafe-au-lait spots and fibromatous tumors of the skin. In this article, we report the overlap of neurofibromatosis-1 with Turner and Klinefelter syndromes. Thus, these disorders might overlap within the same patient. Due to these cases, we suggest that each patient with Turner-like symptoms or Klinefelter's-like syndrome, be carefully examined for café au lait macules before the initiation of hormone replacement treatment.

  6. Intermittency Models and Spot Measurements

    NASA Technical Reports Server (NTRS)

    Ashworth, D. A.

    2007-01-01

    Experimental work at the University of Oxford Osney Lab has demonstrated characteristics of the late-stage transition process by the use of thin-film heat transfer gauges. The development of turbulent spots has been observed in a range of environments, including flat plates, turbine blade cascade tests and wake-passing experiments. These results were taken at Mach/Reynolds numbers and gas-to-wall temperature ratios representative of gas turbines. Analyses of the spot characteristics are consistent with measurements taken in low speed experiments, and support the Schubauer and Klebanoff type of turbulent spots. The addition of simulated wakes from upstream stages has been observed to be primarily superpositional for these tests.

  7. Poisson's spot and Gouy phase

    NASA Astrophysics Data System (ADS)

    da Paz, I. G.; Soldati, Rodolfo; Cabral, L. A.; de Oliveira, J. G. G.; Sampaio, Marcos

    2016-12-01

    Recently there have been experimental results on Poisson spot matter-wave interferometry followed by theoretical models describing the relative importance of the wave and particle behaviors for the phenomenon. We propose an analytical theoretical model for Poisson's spot with matter waves based on the Babinet principle, in which we use the results for free propagation and single-slit diffraction. We take into account effects of loss of coherence and finite detection area using the propagator for a quantum particle interacting with an environment. We observe that the matter-wave Gouy phase plays a role in the existence of the central peak and thus corroborates the predominantly wavelike character of the Poisson's spot. Our model shows remarkable agreement with the experimental data for deuterium (D2) molecules.

  8. Laser based spot weld characterization

    NASA Astrophysics Data System (ADS)

    Jonietz, Florian; Myrach, Philipp; Rethmeier, Michael; Suwala, Hubert; Ziegler, Mathias

    2016-02-01

    Spot welding is one of the most important joining technologies, especially in the automotive industry. Hitherto, the quality of spot welded joints is tested mainly by random destructive tests. A nondestructive testing technique offers the benefit of cost reduction of the testing procedure and optimization of the fabrication process, because every joint could be examined. This would lead to a reduced number of spot welded joints, as redundancies could be avoided. In the procedure described here, the spot welded joint between two zinc-coated steel sheets (HX340LAD+Z100MB or HC340LA+ZE 50/50) is heated optically on one side. Laser radiation and flash light are used as heat sources. The melted zone, the so called "weld nugget" provides the mechanical stability of the connection, but also constitutes a thermal bridge between the sheets. Due to the better thermal contact, the spot welded joint reveals a thermal behavior different from the surrounding material, where the heat transfer between the two sheets is much lower. The difference in the transient thermal behavior is measured with time resolved thermography. Hence, the size of the thermal contact between the two sheets is determined, which is directly correlated to the size of the weld nugget, indicating the quality of the spot weld. The method performs well in transmission with laser radiation and flash light. With laser radiation, it works even in reflection geometry, thus offering the possibility of testing with just one-sided accessibility. By using heating with collimated laser radiation, not only contact-free, but also remote testing is feasible. A further convenience compared to similar thermographic approaches is the applicability on bare steel sheets without any optical coating for emissivity correction. For this purpose, a proper way of emissivity correction was established.

  9. Rocky Mountain spotted fever, Panama.

    PubMed

    Estripeaut, Dora; Aramburú, María Gabriela; Sáez-Llorens, Xavier; Thompson, Herbert A; Dasch, Gregory A; Paddock, Christopher D; Zaki, Sherif; Eremeeva, Marina E

    2007-11-01

    We describe a fatal pediatric case of Rocky Mountain spotted fever in Panama, the first, to our knowledge, since the 1950s. Diagnosis was established by immunohistochemistry, PCR, and isolation of Rickettsia rickettsii from postmortem tissues. Molecular typing demonstrated strong relatedness of the isolate to strains of R. rickettsii from Central and South America.

  10. Multiple café au lait spots in familial patients with MAP2K2 mutation.

    PubMed

    Takenouchi, Toshiki; Shimizu, Atsushi; Torii, Chiharu; Kosaki, Rika; Takahashi, Takao; Saya, Hideyuki; Kosaki, Kenjiro

    2014-02-01

    Recent advances in genetic diagnostic technologies have made the classic disease nosology highly complicated. This situation is exemplified by rasopathies, among which neurofibromatosis type 1 and Noonan syndrome represent prototypic entities. The former condition is characterized by multiple café au lait spots and neurofibromas, while the latter is characterized by distinct facial features, webbed neck, congenital heart disease, and a short stature. On rare occasions, the features of both neurofibromatosis and Noonan syndrome co-exist within an individual; such patients are diagnosed as having neurofibromatosis-Noonan syndrome. Here, we report familial patients with multiple café au lait spots and Noonan syndrome-like facial features. A mutation analysis unexpectedly revealed a mutation in MAP2K2 in both the propositus and his mother. The propositus fulfilled the diagnostic criteria for neurofibromatosis type 1, but his mother did not. Their phenotype was not consistent with that of cardio-facio-cutaneous syndrome, which is classically known to be associated with MAP2K2 mutations. The mother of the propositus had cervical cancer at the age of 23 years, consistent with the oncogenic tendency associated with rasopathies. The phenotypic combination of multiple café au lait spots and Noonan syndrome-like facial features suggested a diagnosis of neurofibromatosis-Noonan syndrome. Whether this condition represents a discrete disease entity or a variable expression of neurofibromatosis type 1 has long been debated. The present observation suggests that some perturbation in the RAS/MAPK signaling cascade results in multiple café au lait spots, a key diagnostic phenotype of rasopathies, although the exact mechanism remains to be elucidated.

  11. 9 CFR 149.4 - Spot audit.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Spot audit. 149.4 Section 149.4... LIVESTOCK IMPROVEMENT VOLUNTARY TRICHINAE CERTIFICATION PROGRAM § 149.4 Spot audit. (a) In addition to regularly scheduled site audits, certified production sites will be subject to spot audits. (1) Random...

  12. 9 CFR 149.4 - Spot audit.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Spot audit. 149.4 Section 149.4... LIVESTOCK IMPROVEMENT VOLUNTARY TRICHINAE CERTIFICATION PROGRAM § 149.4 Spot audit. (a) In addition to regularly scheduled site audits, certified production sites will be subject to spot audits. (1) Random...

  13. Managing Rocky Mountain spotted fever.

    PubMed

    Minniear, Timothy D; Buckingham, Steven C

    2009-11-01

    Rocky Mountain spotted fever is caused by the tick-borne bacterium Rickettsia rickettsii. Symptoms range from moderate illness to severe illness, including cardiovascular compromise, coma and death. The disease is prevalent in most of the USA, especially during warmer months. The trademark presentation is fever and rash with a history of tick bite, although tick exposure is unappreciated in over a third of cases. Other signature symptoms include headache and abdominal pain. The antibiotic therapy of choice for R. rickettsii infection is doxycycline. Preventive measures for Rocky Mountain spotted fever and other tick-borne diseases include: wearing long-sleeved, light colored clothing; checking for tick attachment and removing attached ticks promptly; applying topical insect repellent; and treating clothing with permethrin.

  14. Sweet Spots and Door Stops

    ERIC Educational Resources Information Center

    Thompson, Michael; Tsui, Stella; Leung, Chi Fan

    2011-01-01

    A sweet spot is referred to in sport as the perfect place to strike a ball with a racquet or bat. It is the point of contact between bat and ball where maximum results can be produced with minimal effort from the hand of the player. Similar physics can be applied to the less inspiring examples of door stops; the perfect position of a door stop is…

  15. Justifications shape ethical blind spots.

    PubMed

    Pittarello, Andrea; Leib, Margarita; Gordon-Hecker, Tom; Shalvi, Shaul

    2015-06-01

    To some extent, unethical behavior results from people's limited attention to ethical considerations, which results in an ethical blind spot. Here, we focus on the role of ambiguity in shaping people's ethical blind spots, which in turn lead to their ethical failures. We suggest that in ambiguous settings, individuals' attention shifts toward tempting information, which determines the magnitude of their lies. Employing a novel ambiguous-dice paradigm, we asked participants to report the outcome of the die roll appearing closest to the location of a previously presented fixation cross on a computer screen; this outcome would determine their pay. We varied the value of the die second closest to the fixation cross to be either higher (i.e., tempting) or lower (i.e., not tempting) than the die closest to the fixation cross. Results of two experiments revealed that in ambiguous settings, people's incorrect responses were self-serving. Tracking participants' eye movements demonstrated that people's ethical blind spots are shaped by increased attention toward tempting information.

  16. Systematic Propulsion Optimization Tools (SPOT)

    NASA Technical Reports Server (NTRS)

    Bower, Mark; Celestian, John

    1992-01-01

    This paper describes a computer program written by senior-level Mechanical Engineering students at the University of Alabama in Huntsville which is capable of optimizing user-defined delivery systems for carrying payloads into orbit. The custom propulsion system is designed by the user through the input of configuration, payload, and orbital parameters. The primary advantages of the software, called Systematic Propulsion Optimization Tools (SPOT), are a user-friendly interface and a modular FORTRAN 77 code designed for ease of modification. The optimization of variables in an orbital delivery system is of critical concern in the propulsion environment. The mass of the overall system must be minimized within the maximum stress, force, and pressure constraints. SPOT utilizes the Design Optimization Tools (DOT) program for the optimization techniques. The SPOT program is divided into a main program and five modules: aerodynamic losses, orbital parameters, liquid engines, solid engines, and nozzles. The program is designed to be upgraded easily and expanded to meet specific user needs. A user's manual and a programmer's manual are currently being developed to facilitate implementation and modification.

  17. Downstream Development of a Laminar Spot

    NASA Astrophysics Data System (ADS)

    Sekiya, Naoki; Matsumoto, Akira

    It was well-known that a disturbance, introduced artificially into a supercritical laminar boundary layer along a flat plate, is still laminar in the initial stage of its downstream development. Thus, we named it a "laminar spot" because it resembles a turbulent spot though its velocity perturbation remains laminar. From velocity measurements using a rake-type 16-channel hot-wire probe, we found that in the first stage of the downstream development of a laminar spot, its maximum width was at 0.2δ (what is called the critical layer) and one-half of its lateral growth angle was about 5°, which is almost one-half that of a turbulent spot. We call this region a "laminar spot region". In the present study, we measured in detail the velocity field of a laminar spot using a new hot-wire probe in the laminar spot region. The results showed that a laminar spot consists of some hairpin vortices and some induced U-shaped vortices under the hairpin vortices. Because of the interaction of the velocities induced by the respective vortex legs, the legs of the U-shaped vortices were located at the outermost part of the spot. Moreover, the new vortex legs extended spanwise at about 4° as the spot traveled downstream. Consequently, we concluded that the laminar spot grew spanwise in accordance with the span of these vortex legs.

  18. ESA uncovers Geminga's `hot spot'

    NASA Astrophysics Data System (ADS)

    2004-07-01

    16 July 2004 Astronomers using ESA’s X-ray observatory XMM-Newton have detected a small, bright ‘hot spot’ on the surface of the neutron star called Geminga, 500 light-years away. The hot spot is the size of a football field and is caused by the same mechanism producing Geminga’s X-ray tails. This discovery identifies the missing link between the X-ray and gamma-ray emission from Geminga. hi-res Size hi-res: 1284 kb Credits: ESA, P. Caraveo (IASF, Milan) Geminga's hot spot This figure shows the effects of charged particles accelerated in the magnetosphere of Geminga. Panel (a) shows an image taken with the EPIC instrument on board the XMM-Newton observatory. The bright tails, made of particles kicked out by Geminga’s strong magnetic field, trail the neutron star as it moves about in space. Panel (b) shows how electrically charged particles interact with Geminga’s magnetic field. For example, if electrons (blue) are kicked out by the star, positrons (in red) hit the star’s magnetic poles like in an ‘own goal’. Panel (c) illustrates the size of Geminga’s magnetic field (blue) compared to that of the star itself at the centre (purple). The magnetic field is tilted with respect to Geminga’s rotation axis (red). Panel (d) shows the magnetic poles of Geminga, where charged particles hit the surface of the star, creating a two-million degrees hot spot, a region much hotter than the surroundings. As the star spins on its rotation axis, the hot spot comes into view and then disappears, causing the periodic colour change seen by XMM-Newton. An animated version of the entire sequence can be found at: Click here for animated GIF [low resolution, animated GIF, 5536 KB] Click here for AVI [high resolution, AVI with DIVX compression, 19128 KB] hi-res Size hi-res: 371 kb Credits: ESA, P. Caraveo (IASF, Milan) Geminga's hot spot, panel (a) Panel (a) shows an image taken with the EPIC instrument on board the XMM-Newton observatory. The bright tails, made of

  19. Watermarking spot colors in packaging

    NASA Astrophysics Data System (ADS)

    Reed, Alastair; Filler, TomáÅ.¡; Falkenstern, Kristyn; Bai, Yang

    2015-03-01

    In January 2014, Digimarc announced Digimarc® Barcode for the packaging industry to improve the check-out efficiency and customer experience for retailers. Digimarc Barcode is a machine readable code that carries the same information as a traditional Universal Product Code (UPC) and is introduced by adding a robust digital watermark to the package design. It is imperceptible to the human eye but can be read by a modern barcode scanner at the Point of Sale (POS) station. Compared to a traditional linear barcode, Digimarc Barcode covers the whole package with minimal impact on the graphic design. This significantly improves the Items per Minute (IPM) metric, which retailers use to track the checkout efficiency since it closely relates to their profitability. Increasing IPM by a few percent could lead to potential savings of millions of dollars for retailers, giving them a strong incentive to add the Digimarc Barcode to their packages. Testing performed by Digimarc showed increases in IPM of at least 33% using the Digimarc Barcode, compared to using a traditional barcode. A method of watermarking print ready image data used in the commercial packaging industry is described. A significant proportion of packages are printed using spot colors, therefore spot colors needs to be supported by an embedder for Digimarc Barcode. Digimarc Barcode supports the PANTONE spot color system, which is commonly used in the packaging industry. The Digimarc Barcode embedder allows a user to insert the UPC code in an image while minimizing perceptibility to the Human Visual System (HVS). The Digimarc Barcode is inserted in the printing ink domain, using an Adobe Photoshop plug-in as the last step before printing. Since Photoshop is an industry standard widely used by pre-press shops in the packaging industry, a Digimarc Barcode can be easily inserted and proofed.

  20. Vibrio sp. causing Porites ulcerative white spot disease.

    PubMed

    Arboleda, Mark D M; Reichardt, Wolfgang T

    2010-06-11

    The causative agent of the Indo-Pacific coral disease, Porites ulcerative white spot syndrome (PUWS), that affects Porites spp. and a few other coral genera has so far remained unidentified. Inoculation of thiosulphate citrate bile sucrose (TCBS) agar with tissue material from Porites cylindrica infected with white spot produced colonies of approximately 3 mm diameter consisting of Gram-negative, motile, non-sucrose-fermenting, slightly curved rods with a minimum NaCl requirement of 0.3%. Three of these putative Vibrio sp. isolates were used for infection trials that included different stages of cell growth. Four modes of inoculation and 3 stages of bacterial cell growth were considered for testing Koch's postulates. Stationary phase cells proved more consistently infectious than did exponentially growing or starved cells using a 1-step immersion technique at cell concentrations of 10(4) cells ml(-1). A 1-step immersion technique proved more reliable in producing signs of white spot than did other techniques, such as injection, smearing and 2-step immersion of the inoculum. At inoculum densities >10(4) cells ml(-1) further signs of disease, such as tissue degradation and bleaching, also became evident. At elevated temperatures (>29 degrees C) bleaching remained absent for at least 2 mo from non-inoculated corals serving as controls, but was observed in artificially infected coral fragments. Of the 9 seawater aquaria containing healthy specimens of P. cylindrica, 6 showed signs of white spot 15 d after infection with an isolate tentatively identified as Vibrio sp. Based on 99% similarity of its 16S rRNA gene sequence and selected phenotypical features, this isolate revealed a close relationship to V. natriegens and V. parahaemolyticus.

  1. Nonbright spot AVO: Two examples

    SciTech Connect

    Ross, C.P.; Kinman, D.L.

    1994-12-31

    Utilization of amplitude-versus-offset (AVO) attribute sections such as the product of the normal incidence trace and gradient trace have been used extensively in bright spot (Class 3) AVO analysis and interpretation. However, while these sections have often worked well with Class 3 responses they are not reliable indicators of non-bright spot (Class 2) seismic anomalies. Analyzing Class 2 seismic data with AVO products will: (1) not detect the gas-charged reservoir because of near-zero acoustic impedance contrast between the sands and encasing shales, or (2) yield an incorrect (negative) AVO product if the normal incidence and gradient values are opposite in sign. Class 2 offset responses are divided into two sub-categories: those with phase reversals (Class 2p) and those without phase reversals (Class 2). An AVO procedure for these types of Class 2 anomalies is presented through two examples. The technique better exploits the nature of the prestack response, yielding a more definitive AVO attribute section, and the technique is adaptive to both Class 2 and Class 2p responses. When compared to a conventionally processed relative amplitude seismic section with characteristically low amplitude responses for near-zero acoustic impedance sands, this procedure clearly denotes the presence of gas-charged pore fluids within the reservoir.

  2. Still from Red Spot Movie

    NASA Technical Reports Server (NTRS)

    2000-01-01

    This image is one of seven from the narrow-angle camera on NASA's Cassini spacecraft assembled as a brief movie of cloud movements on Jupiter. It was taken with a blue filter. The smallest features visible are about 500 kilometers (about 300 miles) across.

    Small bright clouds appear suddenly to the west of the Great Red Spot. Based on data from NASA's Galileo spacecraft, scientists suspect that these small white features are lightning storms, where falling raindrops create an electrical charge. The lightning storms eventually merge with the Red Spot and surrounding jets, and may be the main energy source for these large-scale features. Imaging observations of the darkside of the planet in the weeks following Cassini's closest approach to Jupiter on Dec. 30, 2000 will search for lightning storms like these.

    This image was re-projected by cylindrical-map projection of an image taken in the first week of October 2000. It shows an area from 50 degrees north of Jupiter's equator to 50 degrees south, extending 100 degrees east west, about one quarter of Jupiter's circumference.

    Cassini is a cooperative project of NASA, the European Space Agency and the Italian Space Agency. The Jet Propulsion Laboratory, a division of the California Institute of Technology in Pasadena, manages the Cassini mission for NASA's Office of Space Science, Washington, D.C.

  3. Spot and Runway Departure Advisor

    NASA Technical Reports Server (NTRS)

    Jung, Yoon Chul

    2013-01-01

    The Spot and Runway Departure Advisor (SARDA) is a research prototype of a decision support tool for ATC tower controllers to assist in manging and controlling traffic on the surface of an airport. SARDA employs a scheduler to generate an optimal runway schedule and gate push-back - spot release sequence and schedule that improves efficiency of surface operations. The advisories for ATC tower controllers are displayed on an Electronic Flight Strip (EFS) system. The human-in-the-loop simulation of the SARDA tool was conducted for east operations of Dallas-Ft. Worth International Airport (DFW) to evaluate performance of the SARDA tool and human factors, such as situational awareness and workload. The results indicates noticeable taxi delay reduction and fuel savings by using the SARDA tool. Reduction in controller workload were also observed throughout the scenario runs. The future plan includes modeling and simulation of the ramp operations of the Charlotte International Airport, and develop a decision support tool for the ramp controllers.

  4. Instructor Debrief Training in SPOT

    NASA Technical Reports Server (NTRS)

    Martin, Lynne; Orasanu, Judith; Villeda, Eric; Conners, Mary M. (Technical Monitor)

    2002-01-01

    One way to enhance the effectiveness of Special Purpose Operational Training' (SPOT) debriefing sessions may be for instructors to make explicit connections between the Crew Resource Management (CRM) concepts a carrier advocates and the behaviors displayed by the crew in question. A tool listing key behaviors from the scenario was devised, accompanied by an instructors' training session in which links were made between the behaviors and the underlying CRM processes they reflect. The aim of the tool is to assist instructors to focus the debriefing on the key SPOT/ CRM issues, in this case on planning. A second tool suggested ways to facilitate the discussion. Fourteen instructors at a major U.S. carrier took part in the training session and used the toolkit in their subsequent debriefs. Pre- and post-training debriefing samples from each instructor were compared to assess whether there were any changes in instructors' approaches to discussions in terms of the topics they covered and how they raised the points.

  5. Solar 'hot spots' are still hot

    NASA Technical Reports Server (NTRS)

    Bai, Taeil

    1990-01-01

    Longitude distributions of solar flares are not random but show evidence for active zones (or hot spots) where flares are concentrated. According to a previous study, two hot spots in the northern hemisphere, which rotate with a synodic period of about 26.72 days, produced the majority of major flares, during solar cycles 20 and 21. The more prominent of these two hot spots is found to be still active during the rising part of cycle 22, producing the majority of northern hemisphere major flares. The synodic rotation period of this hot spot is 26.727 + or - 0.007 days. There is also evidence for hot spots in the southern hemisphere. Two hot spots separated by 180 deg are found to rotate with a period of 29.407 days, with one of them having persisted in the same locations during cycles 19-22 and the other, during cycles 20-22.

  6. Spotted bass Micropterus punctulatus (Rafinesque 1819)

    USGS Publications Warehouse

    Churchill, Timothy N.; Bettoli, Phillip William

    2015-01-01

    Three subspecies of Spotted Bass Micropterus punctulatus were historically recognized: the smaller Northern Spotted Bass M. p. punctulatus, the larger, longer-lived Alabama Spotted Bass M. p. henshalli, and the now invalidated Wichita Spotted Bass M. p. wichitae (Bailey and Hubbs 1940; Cofer 1995; Warren 2009; Rider and Maceina 2015, this volume). The subspecific status has been examined over the past decade as advanced genetic analyses have been developed (e.g., Kassler et al. 2002; Baker et al. 2008; Tringali et al. 2015, this volume). The American Fisheries Society has recently changed the designation of the Alabama Spotted Bass to a separate species, Alabama Bass M. henshalli (Page et al. 2013). The remainder of this paper will discuss the biology and conservation of only Spotted Bass. Both species have been observed to hybridize with other Micropterus spp. (Koppelman 1994; Pierce and Van Den Avyle 1997; Barwick et al. 2006).

  7. Integrating sustainable hunting in biodiversity protection in Central Africa: hot spots, weak spots, and strong spots.

    PubMed

    Fa, John E; Olivero, Jesús; Farfán, Miguel Ángel; Márquez, Ana Luz; Vargas, Juan Mario; Real, Raimundo; Nasi, Robert

    2014-01-01

    Wild animals are a primary source of protein (bushmeat) for people living in or near tropical forests. Ideally, the effect of bushmeat harvests should be monitored closely by making regular estimates of offtake rate and size of stock available for exploitation. However, in practice, this is possible in very few situations because it requires both of these aspects to be readily measurable, and even in the best case, entails very considerable time and effort. As alternative, in this study, we use high-resolution, environmental favorability models for terrestrial mammals (N = 165) in Central Africa to map areas of high species richness (hot spots) and hunting susceptibility. Favorability models distinguish localities with environmental conditions that favor the species' existence from those with detrimental characteristics for its presence. We develop an index for assessing Potential Hunting Sustainability (PHS) of each species based on their ecological characteristics (population density, habitat breadth, rarity and vulnerability), weighted according to restrictive and permissive assumptions of how species' characteristics are combined. Species are classified into five main hunting sustainability classes using fuzzy logic. Using the accumulated favorability values of all species, and their PHS values, we finally identify weak spots, defined as high diversity regions of especial hunting vulnerability for wildlife, as well as strong spots, defined as high diversity areas of high hunting sustainability potential. Our study uses relatively simple models that employ easily obtainable data of a species' ecological characteristics to assess the impacts of hunting in tropical regions. It provides information for management by charting the geography of where species are more or less likely to be at risk of extinction from hunting.

  8. [Syndromes 2. Pfeiffer syndrome].

    PubMed

    Freihofer, H P

    1998-07-01

    Acrocephalosyndactylias are syndromes characterized by abnormalities of the head (craniosynostosis), the face (hypertelorism, retromaxillism), hands and feet (cutaneous or bony syndactyly). Inheritance is autosomal dominant, but spontaneous cases are described also. The group is divided into several syndromes with varying penetrance and expressivity. As an example of an acrocephalosyndactylia is the Pfeiffer syndrome presented.

  9. Voyager 1 Red Spot Movie

    NASA Technical Reports Server (NTRS)

    2000-01-01

    This movie shows the portion of Jupiter around the Great Red Spot as it swirls through more than 60 Jupiter days. Notice the difference in speed and direction of the various zones of the atmosphere. The interaction of the atmospheric clouds and storm shows how dynamic the Jovian atmosphere is.

    As Voyager 1 approached Jupiter in 1979, it took images of the planet at regular intervals. This sequence is made from 66 images taken once every Jupiter rotation period (about 10 hours). This time-lapse movie uses images taken every time Jupiter longitude 68W passed under the spacecraft. These images were acquired in the Blue filter from Jan. 6 to Feb. 3 1979. The spacecraft flew from 58 million kilometers to 31 million kilometers from Jupiter during that time.

    This time-lapse movie was produced at JPL by the Image Processing Laboratory in 1979.

  10. Fatal Rocky Mountain spotted fever.

    PubMed

    Hattwick, M A; Retailliau, H; O'Brien, R J; Slutzker, M; Fontaine, R E; Hanson, B

    1978-09-29

    Forty-four fatal cases of Rocky Mountain spotted fever (RMSF) occurring in 1974 were compared with 50 nonfatal cases of similar age, sex, date of onset, and place of occurrence. Diagnosis and initiation of treatment in fatal cases were substantially delayed compared with nonfatal cases. Several reasons for this delay were identified: (1) the rash appeared later in the course of illness in the fatal cases, often not until the patient was terminal, (2) a history of tick bite was less often obtained during life or obtained late in the clinical course in fatal cases, and (3) initial nonspecific symptoms or unexpected symptoms led to an initial diagnosis of more common diseases. Only two fatal cases were treated with either tetracycline or chloramphenicol before the sixth day of illness. Presumptive diagnosis of RMSF and initiation of tetracycline therapy before onset of rash may be necessary to reduce mortality.

  11. SU-E-T-510: Interplay Between Spots Sizes, Spot / Line Spacing and Motion in Spot Scanning Proton Therapy

    SciTech Connect

    Lee, TK

    2015-06-15

    Purpose In proton beam configuration for spot scanning proton therapy (SSPT), one can define the spacing between spots and lines of scanning as a ratio of given spot size. If the spacing increases, the number of spots decreases which can potentially decrease scan time, and so can whole treatment time, and vice versa. However, if the spacing is too large, the uniformity of scanned field decreases. Also, the field uniformity can be affected by motion during SSPT beam delivery. In the present study, the interplay between spot/ line spacing and motion is investigated. Methods We used four Gaussian-shape spot sizes with 0.5cm, 1.0cm, 1.5cm, and 2.0cm FWHM, three spot/line spacing that creates uniform field profile which are 1/3*FWHM, σ/3*FWHM and 2/3*FWHM, and three random motion amplitudes within, +/−0.3mm, +/−0.5mm, and +/−1.0mm. We planned with 2Gy uniform single layer of 10×10cm2 and 20×20cm2 fields. Then, mean dose within 80% area of given field size, contrubuting MU per each spot assuming 1cGy/MU calibration for all spot sizes, number of spots and uniformity were calculated. Results The plans with spot/line spacing equal to or smaller than 2/3*FWHM without motion create ∼100% uniformity. However, it was found that the uniformity decreases with increased spacing, and it is more pronounced with smaller spot sizes, but is not affected by scanned field sizes. Conclusion It was found that the motion during proton beam delivery can alter the dose uniformity and the amount of alteration changes with spot size which changes with energy and spot/line spacing. Currently, robust evaluation in TPS (e.g. Eclipse system) performs range uncertainty evaluation using isocenter shift and CT calibration error. Based on presented study, it is recommended to add interplay effect evaluation to robust evaluation process. For future study, the additional interplay between the energy layers and motion is expected to present volumetric effect.

  12. Ambystoma maculatum (spotted salamander). Reproduction

    USGS Publications Warehouse

    Glorioso, Brad M.; Waddle, Hardin; Hefner, Jeromi

    2012-01-01

    The Spotted Salamander is a wide-ranging salamander of the eastern United States that typically breeds in winter or early spring in ephemeral pools in lowland forests. Ambystoma maculatum is known to deposit 2-4 egg masses per year, each containing 1-250 eggs. As part of ongoing research into the ecology and reproductive biology of Spotted Salamanders in the Kisatchie District of Kisatchie National Forest in Natchitoches Parish, Louisiana, USA, we have been counting the number of embryos per egg mass. We captured seven female A. maculatum in a small pool, six of which were still gravid. We took standard measurements, including SVL, and then implanted a Passive Integrated Transponder (PIT tag) into each adult female as was the protocol. About an hour after processing these animals we marked new A. maculatum egg masses found in the same small pool using PVC pin flags pushed carefully through the outer jelly. We did not have enough time to process them that evening, and it was not until a few days later that we photographed those masses. We discovered that one of the masses contained a PIT tag in the outer jelly that corresponded to one of the six gravid females that were marked that same evening. To our knowledge, this is the first report of PIT tags being the means, albeit coincidentally, by which a particular egg mass of Ambystoma maculatum has been assigned to a particular female. For our purposes, losing the PIT tag from the adult female is counter to the goals of our study of this population, and we will no longer be implanting PIT tags into gravid females.

  13. Strategic Improvements to TSA Spot Program

    DTIC Science & Technology

    2015-03-01

    Administration’s Screening Passengers by Observation Techniques (SPOT) program analyzes the Government Accountability Office (GAO) and Office of Inspector...INTENTIONALLY LEFT BLANK iv ABSTRACT This study of Transportation Security Administration’s Screening Passengers by Observation Techniques ...area SME subject matter expert SPOT screening passengers by observation techniques TAC threat assessment capabilities TAD threat assessment

  14. Hot Spot Removal System: System description

    SciTech Connect

    1997-09-01

    Hazardous wastes contaminated with radionuclides, chemicals, and explosives exist across the Department of Energy complex and need to be remediated due to environmental concerns. Currently, an opportunity is being developed to dramatically reduce remediation costs and to assist in the acceleration of schedules associated with these wastes by deploying a Hot Spot Removal System. Removing the hot spot from the waste site will remove risk driver(s) and enable another, more cost effective process/option/remedial alternative (i.e., capping) to be applied to the remainder of the site. The Hot Spot Removal System consists of a suite of technologies that will be utilized to locate and remove source terms. Components of the system can also be used in a variety of other cleanup activities. This Hot Spot Removal System Description document presents technologies that were considered for possible inclusion in the Hot Spot Removal System, technologies made available to the Hot Spot Removal System, industrial interest in the Hot Spot Removal System`s subsystems, the schedule required for the Hot Spot Removal System, the evaluation of the relevant technologies, and the recommendations for equipment and technologies as stated in the Plan section.

  15. HotSpot Software Test Plan

    SciTech Connect

    Walker, H; Homann, S G

    2009-03-12

    This Software Test Plan (STP) describes the procedures used to verify and validate that the HotSpot Health Physics Codes meet the requirements of its user base, which includes: (1) Users of the PC version of HotSpot conducting consequence assessment, hazard assessment and safety analysis calculations; and (2) Users of the NARAC Web and iClient software tools, which allow users to run HotSpot for consequence assessment modeling. This plan is intended to meet Critical Recommendation 2 from the Software Evaluation of HotSpot and DOE Safety Software Toolbox Recommendation for inclusion of HotSpot in the Department of Energy (DOE) Safety Software Toolbox. These users and sponsors of the HotSpot software and the organizations they represent constitute the intended audience for this document. HotSpot software is maintained for the Department of Energy Office of Emergency Operations by the National Atmospheric Release Advisory Center (NARAC) at Lawrence Livermore National Laboratory (LLNL). An overview of HotSpot and NARAC are provided.

  16. Rocky Mountain Spotted Fever in Argentina

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We describe the first molecular confirmation of Rickettsia rickettsii, the cause of Rocky Mountain spotted fever (RMSF), from a tick vector, Amblyomma cajennense, and from a cluster of fatal spotted fever cases in Argentina. Questing A. cajennense ticks were collected at or near sites of presumed or...

  17. Rocky Mountain spotted fever in children.

    PubMed

    Woods, Charles R

    2013-04-01

    Rocky Mountain spotted fever is typically undifferentiated from many other infections in the first few days of illness. Treatment should not be delayed pending confirmation of infection when Rocky Mountain spotted fever is suspected. Doxycycline is the drug of choice even for infants and children less than 8 years old.

  18. Rocky Mountain spotted fever in Argentina

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cases of epidemic typhus have been documented in Argentina since 1919; however, no confirmed reports of spotted fever rickettsiosis were described in this country until 1999. We describe the first molecular confirmation of Rickettsia rickettsii, the etiologic agent of Rocky Mountain spotted fever (R...

  19. Rocky Mountain Spotted Fever: Statistics and Epidemiology

    MedlinePlus

    ... Search The CDC Cancel Submit Search The CDC Rocky Mountain Spotted Fever (RMSF) Note: Javascript is disabled or is not ... message, please visit this page: About CDC.gov . Rocky Mountain Spotted Fever (RMSF) Symptoms, Diagnosis, and Treatment Statistics and Epidemiology ...

  20. Habitat Suitability Index Models: Spotted seatrout

    USGS Publications Warehouse

    Kostecki, Paul T.

    1984-01-01

    The estuarine spotted seatrout, a primarily estuarine species, is one of the most important sport and commercial fishes in coastal Gulf of Mexico waters (Arnold et a1. 1976). Spotted seatrout rank second by weight in catches by U.S. saltwater sport fishermen (National Marine Fisheries Services 1981) .

  1. Moebius Syndrome

    MedlinePlus

    ... children with Moebius syndrome have some degree of autism. There are four recognized categories of Moebius syndrome: ... children with Moebius syndrome have some degree of autism. There are four recognized categories of Moebius syndrome: ...

  2. 7 CFR 28.415 - Low Middling Light Spotted Color.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 2 2014-01-01 2014-01-01 false Low Middling Light Spotted Color. 28.415 Section 28... Spotted Color. Low Middling Light Spotted Color is color which in spot or color, or both, is between Low Middling Color and Low Middling Spotted Color....

  3. 7 CFR 28.415 - Low Middling Light Spotted Color.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 2 2013-01-01 2013-01-01 false Low Middling Light Spotted Color. 28.415 Section 28... Spotted Color. Low Middling Light Spotted Color is color which in spot or color, or both, is between Low Middling Color and Low Middling Spotted Color....

  4. 7 CFR 28.415 - Low Middling Light Spotted Color.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Low Middling Light Spotted Color. 28.415 Section 28... Spotted Color. Low Middling Light Spotted Color is color which in spot or color, or both, is between Low Middling Color and Low Middling Spotted Color....

  5. 7 CFR 28.413 - Middling Light Spotted Color.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 2 2013-01-01 2013-01-01 false Middling Light Spotted Color. 28.413 Section 28.413... Spotted Color. Middling Light Spotted Color is color which in spot or color, or both, is between Middling Color and Middling Spotted Color....

  6. 7 CFR 28.411 - Good Middling Light Spotted Color.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 2 2011-01-01 2011-01-01 false Good Middling Light Spotted Color. 28.411 Section 28... Light Spotted Color. Good Middling Light Spotted Color is color which in spot or color, or both, is between Good Middling Color and Good Middling Spotted Color....

  7. 7 CFR 28.415 - Low Middling Light Spotted Color.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 2 2011-01-01 2011-01-01 false Low Middling Light Spotted Color. 28.415 Section 28... Spotted Color. Low Middling Light Spotted Color is color which in spot or color, or both, is between Low Middling Color and Low Middling Spotted Color....

  8. 7 CFR 28.412 - Strict Middling Light Spotted Color.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 2 2013-01-01 2013-01-01 false Strict Middling Light Spotted Color. 28.412 Section 28... Light Spotted Color. Strict Middling Light Spotted Color is color which in spot or color, or both, is between Strict Middling Color and Strict Middling Spotted Color....

  9. 7 CFR 28.415 - Low Middling Light Spotted Color.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 2 2012-01-01 2012-01-01 false Low Middling Light Spotted Color. 28.415 Section 28... Spotted Color. Low Middling Light Spotted Color is color which in spot or color, or both, is between Low Middling Color and Low Middling Spotted Color....

  10. 7 CFR 28.411 - Good Middling Light Spotted Color.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 2 2012-01-01 2012-01-01 false Good Middling Light Spotted Color. 28.411 Section 28... Light Spotted Color. Good Middling Light Spotted Color is color which in spot or color, or both, is between Good Middling Color and Good Middling Spotted Color....

  11. 7 CFR 28.412 - Strict Middling Light Spotted Color.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Strict Middling Light Spotted Color. 28.412 Section 28... Light Spotted Color. Strict Middling Light Spotted Color is color which in spot or color, or both, is between Strict Middling Color and Strict Middling Spotted Color....

  12. 7 CFR 28.413 - Middling Light Spotted Color.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 2 2011-01-01 2011-01-01 false Middling Light Spotted Color. 28.413 Section 28.413... Spotted Color. Middling Light Spotted Color is color which in spot or color, or both, is between Middling Color and Middling Spotted Color....

  13. 7 CFR 28.411 - Good Middling Light Spotted Color.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 2 2013-01-01 2013-01-01 false Good Middling Light Spotted Color. 28.411 Section 28... Light Spotted Color. Good Middling Light Spotted Color is color which in spot or color, or both, is between Good Middling Color and Good Middling Spotted Color....

  14. 7 CFR 28.412 - Strict Middling Light Spotted Color.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 2 2012-01-01 2012-01-01 false Strict Middling Light Spotted Color. 28.412 Section 28... Light Spotted Color. Strict Middling Light Spotted Color is color which in spot or color, or both, is between Strict Middling Color and Strict Middling Spotted Color....

  15. 7 CFR 28.413 - Middling Light Spotted Color.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 2 2014-01-01 2014-01-01 false Middling Light Spotted Color. 28.413 Section 28.413... Spotted Color. Middling Light Spotted Color is color which in spot or color, or both, is between Middling Color and Middling Spotted Color....

  16. 7 CFR 28.413 - Middling Light Spotted Color.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 2 2012-01-01 2012-01-01 false Middling Light Spotted Color. 28.413 Section 28.413... Spotted Color. Middling Light Spotted Color is color which in spot or color, or both, is between Middling Color and Middling Spotted Color....

  17. 7 CFR 28.411 - Good Middling Light Spotted Color.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 2 2014-01-01 2014-01-01 false Good Middling Light Spotted Color. 28.411 Section 28... Light Spotted Color. Good Middling Light Spotted Color is color which in spot or color, or both, is between Good Middling Color and Good Middling Spotted Color....

  18. 7 CFR 28.412 - Strict Middling Light Spotted Color.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 2 2014-01-01 2014-01-01 false Strict Middling Light Spotted Color. 28.412 Section 28... Light Spotted Color. Strict Middling Light Spotted Color is color which in spot or color, or both, is between Strict Middling Color and Strict Middling Spotted Color....

  19. 7 CFR 28.411 - Good Middling Light Spotted Color.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Good Middling Light Spotted Color. 28.411 Section 28... Light Spotted Color. Good Middling Light Spotted Color is color which in spot or color, or both, is between Good Middling Color and Good Middling Spotted Color....

  20. 7 CFR 28.412 - Strict Middling Light Spotted Color.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 2 2011-01-01 2011-01-01 false Strict Middling Light Spotted Color. 28.412 Section 28... Light Spotted Color. Strict Middling Light Spotted Color is color which in spot or color, or both, is between Strict Middling Color and Strict Middling Spotted Color....

  1. 7 CFR 28.413 - Middling Light Spotted Color.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Middling Light Spotted Color. 28.413 Section 28.413... REGULATIONS COTTON CLASSING, TESTING, AND STANDARDS Standards Light Spotted Cotton § 28.413 Middling Light Spotted Color. Middling Light Spotted Color is color which in spot or color, or both, is between...

  2. Neptune's small dark spot (D2)

    NASA Technical Reports Server (NTRS)

    1999-01-01

    This bulls-eye view of Neptune's small dark spot (D2) was obtained by Voyager 2's narrow-angle camera. Banding surrounding the feature indicates unseen strong winds, while structures within the bright spot suggest both active upwelling of clouds and rotation about the center. A rotation rate has not yet been measured, but the V-shaped structure near the right edge of the bright area indicates that the spot rotates clockwise. Unlike the Great Red Spot on Jupiter, which rotates counterclockwise, if the D2 spot on Neptune rotates clockwise, the material will be descending in the dark oval region. The fact that infrared data will yield temperature information about the region above the clouds makes this observation especially valuable. The Voyager Mission is conducted by JPL for NASA's Office of Space Science and Applications.

  3. Rocky Mountain spotted fever in Argentina.

    PubMed

    Paddock, Christopher D; Fernandez, Susana; Echenique, Gustavo A; Sumner, John W; Reeves, Will K; Zaki, Sherif R; Remondegui, Carlos E

    2008-04-01

    We describe the first molecular confirmation of Rickettsia rickettsii, the cause of Rocky Mountain spotted fever (RMSF), from a tick vector, Amblyomma cajennense, and from a cluster of fatal spotted fever cases in Argentina. Questing A. cajennense ticks were collected at or near sites of presumed or confirmed cases of spotted fever rickettsiosis in Jujuy Province and evaluated by polymerase chain reaction assays for spotted fever group rickettsiae. DNA of R. rickettsii was amplified from a pool of A. cajennense ticks and from tissues of one of four patients who died during 2003-2004 after illnesses characterized by high fever, severe headache, myalgias, and petechial rash. The diagnosis of spotted fever rickettsiosis was confirmed in the other patients by indirect immunofluorescence antibody and immunohistochemical staining techniques. These findings show the existence of RMSF in Argentina and emphasize the need for clinicians throughout the Americas to consider RMSF in patients with febrile rash illnesses.

  4. Fatal Israeli spotted fever in children.

    PubMed

    Yagupsky, P; Wolach, B

    1993-11-01

    We describe three Israeli children with fatal spotted fever. Clinical disease was characterized by irreversible shock, encephalopathy, renal failure, bleeding tendency, and death within 24 hours of admission. None of the patients had a history of tick bite, and no tache noire was noted. One child presented without rash, and the other two did not have antibodies to spotted-fever-group rickettsiae. The disease was confirmed by isolation of Rickettsia conorii from the patients' blood and tissues in cell cultures or from susceptible laboratory animals inoculated with human specimens. The present cases demonstrate the existence of a severe form of Israeli spotted fever in this population that resembles Rocky Mountain spotted fever. Because Israeli spotted fever may follow a quick, unpredictable, rapidly fatal clinical course, specific antimicrobial therapy should be promptly administered whenever the diagnosis is suspected.

  5. TEVA-SPOT Toolkit 1.2

    SciTech Connect

    Berry, Jonathan; Riesen, Lee Ann; Hart, William

    2007-07-26

    The TEVA-SPOT Toolkit (SPOT) supports the design of contaminant warning systems (CWSs) that use real-time sensors to detect contaminants in municipal water distribution networks. Specifically, SPOT provides the capability to select the locations for installing sensors in order to maximize the utility and effectiveness of the CWS. SPOT models the sensor placement process as an optimization problem, and the user can specify a wide range of performance objectives for contaminant warning system design, including population health effects, time to detection, extent of contamination, volume consumed and number of failed detections. For example, a SPOT user can integrate expert knowledge during the design process by specigying required sensor placements or designating network locations as forbidden. Further, cost considerations can be integrated by limiting the design with user-specified installation costs at each location.

  6. Laser Spot Detection Based on Reaction Diffusion

    PubMed Central

    Vázquez-Otero, Alejandro; Khikhlukha, Danila; Solano-Altamirano, J. M.; Dormido, Raquel; Duro, Natividad

    2016-01-01

    Center-location of a laser spot is a problem of interest when the laser is used for processing and performing measurements. Measurement quality depends on correctly determining the location of the laser spot. Hence, improving and proposing algorithms for the correct location of the spots are fundamental issues in laser-based measurements. In this paper we introduce a Reaction Diffusion (RD) system as the main computational framework for robustly finding laser spot centers. The method presented is compared with a conventional approach for locating laser spots, and the experimental results indicate that RD-based computation generates reliable and precise solutions. These results confirm the flexibility of the new computational paradigm based on RD systems for addressing problems that can be reduced to a set of geometric operations. PMID:26938537

  7. Laser Spot Detection Based on Reaction Diffusion.

    PubMed

    Vázquez-Otero, Alejandro; Khikhlukha, Danila; Solano-Altamirano, J M; Dormido, Raquel; Duro, Natividad

    2016-03-01

    Center-location of a laser spot is a problem of interest when the laser is used for processing and performing measurements. Measurement quality depends on correctly determining the location of the laser spot. Hence, improving and proposing algorithms for the correct location of the spots are fundamental issues in laser-based measurements. In this paper we introduce a Reaction Diffusion (RD) system as the main computational framework for robustly finding laser spot centers. The method presented is compared with a conventional approach for locating laser spots, and the experimental results indicate that RD-based computation generates reliable and precise solutions. These results confirm the flexibility of the new computational paradigm based on RD systems for addressing problems that can be reduced to a set of geometric operations.

  8. Geo Spots and Vortex Theory

    NASA Astrophysics Data System (ADS)

    Straser, Valentino

    2014-05-01

    ), when limited to particular regions, may have created in the past and perhaps still do to this day torsions in localized spots of the Earth's crust (Geo Spots), which over time have conditioned the distribution of tectonic stresses on the surface of the Earth at a regional scale.

  9. HUBBLE FINDS NEW DARK SPOT ON NEPTUNE

    NASA Technical Reports Server (NTRS)

    2002-01-01

    NASA's Hubble Space Telescope has discovered a new great dark spot, located in the northern hemisphere of the planet Neptune. Because the planet's northern hemisphere is now tilted away from Earth, the new feature appears near the limb of the planet. The spot is a near mirror-image to a similar southern hemisphere dark spot that was discovered in 1989 by the Voyager 2 probe. In 1994, Hubble showed that the southern dark spot had disappeared. Like its predecessor, the new spot has high altitude clouds along its edge, caused by gasses that have been pushed to higher altitudes where they cool to form methane ice crystal clouds. The dark spot may be a zone of clear gas that is a window to a cloud deck lower in the atmosphere. Planetary scientists don t know how long lived this new feature might be. Hubble's high resolution will allow astronomers to follow the spot's evolution and other unexpected changes in Neptune's dynamic atmosphere. The image was taken on November 2, 1994 with Hubble's Wide Field Planetary Camera 2, when Neptune was 2.8 billion miles (4.5 billion kilometers) from Earth. Hubble can resolve features as small as 625 miles (1,000 kilometers) across in Neptune's cloud tops. Credit: H. Hammel (Massachusetts Institute of Technology) and NASA

  10. Mediterranean spotted fever and hearing impairment: a rare complication.

    PubMed

    Rossio, Raffaella; Conalbi, Valeria; Castagna, Valentina; Recalcati, Sebastiano; Torri, Adriana; Coen, Massimo; Cassulini, Lucia Restano; Peyvandi, Flora

    2015-06-01

    Mediterranean spotted fever (MSF) is caused by Rickettsia conorii and transmitted by the brown dog tick Rhipicephalus sanguineus. It is prevalent in southern Europe, Africa and central Asia. The disease usually has a benign course and is characterized by fever, myalgia and a characteristic papular rash with an inoculation eschar ('tache noir') at the site of the tick bite. Severe forms of disease can have cardiac, neurologic or renal involvement. Nervous system complications are unusual and may develop in the early phase of disease or as a delayed complication. Neurological symptoms include headache and alterations of the level of consciousness, and some cases of meningoenchefalitis and Guillain-Barrè syndrome have been also reported. Peripheral nerve involvement is reported only in a limited number of case reports. We describe a case of Rickettsia conorii that was complicated with hearing loss and did not respond to specific treatment. Hearing loss is a rare event, but clinicians should be aware of this complication.

  11. Complete Genome Sequence of the Shrimp White Spot Bacilliform Virus

    PubMed Central

    Yang, Feng; He, Jun; Lin, Xionghui; Li, Qin; Pan, Deng; Zhang, Xiaobo; Xu, Xun

    2001-01-01

    We report the first complete genome sequence of a marine invertebrate virus. White spot bacilliform virus (WSBV; or white spot syndrome virus) is a major shrimp pathogen with a high mortality rate and a wide host range. Its double-stranded circular DNA genome of 305,107 bp contains 181 open reading frames (ORFs). Nine homologous regions containing 47 repeated minifragments that include direct repeats, atypical inverted repeat sequences, and imperfect palindromes were identified. This is the largest animal virus that has been completely sequenced. Although WSBV is morphologically similar to insect baculovirus, the two viruses are not detectably related at the amino acid level. Rather, some WSBV genes are more homologous to eukaryotic genes than viral genes. In fact, sequence analysis indicates that WSBV differs from all known viruses, although a few genes display a weak homology to herpesvirus genes. Most of the ORFs encode proteins that bear no homology to any known proteins, either suggesting that WSBV represents a novel class of viruses or perhaps implying a significant evolutionary distance between marine and terrestrial viruses. The most unique feature of WSBV is the presence of an intact collagen gene, a gene encoding an extracellular matrix protein of animal cells that has never been found in any viruses. Determination of the genome of WSBV will facilitate a better understanding of the molecular mechanism underlying the pathogenesis of the WSBV virus and will also provide useful information concerning the evolution and divergence of marine and terrestrial animal viruses at the molecular level. PMID:11689662

  12. Validity of urinary monoamine assay sales under the "spot baseline urinary neurotransmitter testing marketing model".

    PubMed

    Hinz, Marty; Stein, Alvin; Uncini, Thomas

    2011-01-01

    Spot baseline urinary monoamine assays have been used in medicine for over 50 years as a screening test for monoamine-secreting tumors, such as pheochromocytoma and carcinoid syndrome. In these disease states, when the result of a spot baseline monoamine assay is above the specific value set by the laboratory, it is an indication to obtain a 24-hour urine sample to make a definitive diagnosis. There are no defined applications where spot baseline urinary monoamine assays can be used to diagnose disease or other states directly. No peer-reviewed published original research exists which demonstrates that these assays are valid in the treatment of individual patients in the clinical setting. Since 2001, urinary monoamine assay sales have been promoted for numerous applications under the "spot baseline urinary neurotransmitter testing marketing model". There is no published peer-reviewed original research that defines the scientific foundation upon which the claims for these assays are made. On the contrary, several articles have been published that discredit various aspects of the model. To fill the void, this manuscript is a comprehensive review of the scientific foundation and claims put forth by laboratories selling urinary monoamine assays under the spot baseline urinary neurotransmitter testing marketing model.

  13. A Dark Spot on Europa

    NASA Technical Reports Server (NTRS)

    1998-01-01

    This view taken by NASA's Galileo spacecraft of Jupiter's icy moon Europa focuses on a dark, smooth region whose center is the lowest area in this image. To the west (left), it is bounded by a cliff and terraces, which might have been formed by normal faulting. The slopes toward the east (right) leading into the dark spot are gentle.

    Near the center of the dark area, it appears the dark materials have covered some of the bright terrain and ridges. This suggests that when the dark material was deposited, it may have been a fluid or an icy slush.

    Only a few impact craters are visible, with some of them covered or flooded by dark material. Some appear in groups, which may indicate that they are secondary craters formed by debris excavated during a larger impact event. A potential source for these is the nearby crater Mannann`an.

    North is to the top of the picture which is centered at 1 degree south latitude and 225 degrees west longitude. The images in this mosaic have been re-projected to 50 meters (55 yards) per picture element. They were obtained by the Solid State Imaging (SSI) system on March 29, 1998, during Galileo's fourteenth orbit of Jupiter, at ranges as close as 1940 kilometers (1,200 miles) from Europa.

    The Jet Propulsion Laboratory, Pasadena, CA manages the Galileo mission for NASA's Office of Space Science, Washington, DC. JPL is an operating division of California Institute of Technology (Caltech).

    This image and other images and data received from Galileo are posted on the World Wide Web, on the Galileo mission home page at URL http://galileo.jpl.nasa.gov. Background information and educational context for the images can be found at URL http://www.jpl.nasa.gov/galileo/sepo

  14. Finding your innovation sweet spot.

    PubMed

    Goldenberg, Jacob; Horowitz, Roni; Levav, Amnon; Mazursky, David

    2003-03-01

    Most new product ideas are either uninspired or impractical. So how can developers hit the innovation sweet spot--far enough from existing products to attract real interest but close enough that they are feasible to make and market? They can apply five innovation patterns that manipulate existing components of a product and its immediate environment to come up with something both ingenious and viable, say the authors. The subtraction pattern works by removing product components, particularly those that seem desirable or indispensable. Think of the legless high chair that attaches to the kitchen table. The multiplication pattern makes one or more copies of an existing component, then alters those copies in some important way. For example, the Gillette double-bladed razor features a second blade that cuts whiskers at a slightly different angle. By dividing an existing product into its component parts--the division pattern--you can see something that was an integrated whole in an entirely different light. Think of the modern home stereo--it has modular speakers, tuners, and CD and tape players, which allow users to customize their sound systems. The task unification pattern involves assigning a new task to an existing product element or environmental attribute, thereby unifying two tasks in a single component. An example is the defrosting filament in an automobile windshield that also serves as a radio antenna. Finally, the attribute dependency pattern alters or creates the dependent relationships between a product and its environment. For example, by creating a dependent relationship between lens color and external lighting conditions, eyeglass developers came up with a lens that changes color when exposed to sunlight.

  15. Self-Checks Help Spot Melanoma's Return

    MedlinePlus

    ... news/fullstory_163682.html Self-Checks Help Spot Melanoma's Return Patient-detected symptoms were most common way ... Feb. 20, 2017 (HealthDay News) -- Self-checks by melanoma skin cancer patients play an important role in ...

  16. Rocky Mountain Spotted Fever (For Parents)

    MedlinePlus

    ... Safety for the Whole Family Evaluate Your Child's Lyme Disease Risk Lyme Disease Lyme Disease Hey! A Tick Bit Me! Bug Bites and Stings Rocky Mountain Spotted Fever Lyme Disease Contact Us Print Resources Send to a friend ...

  17. Measuring microfocus focal spots using digital radiography

    SciTech Connect

    Fry, David A

    2009-01-01

    Measurement of microfocus spot size can be important for several reasons: (1) Quality assurance during manufacture of microfocus tubes; (2) Tracking performance and stability of microfocus tubes; (3) Determining magnification (especially important for digital radiography where the native spatial resolution of the digital system is not adequate for the application); (4) Knowledge of unsharpness from the focal spot alone. The European Standard EN 12543-5 is based on a simple geometrical method of calculating focal spot size from unsharpness of high magnification film radiographs. When determining microfocus focal spot dimensions using unsharpness measurements both signal-to-noise (SNR) and magnification can be important. There is a maximum accuracy that is a function of SNR and therefore an optimal magnification. Greater than optimal magnification can be used but it will not increase accuracy.

  18. Interferometric diagnostics for magnetic spots detection

    NASA Astrophysics Data System (ADS)

    Ligi, R.

    2014-09-01

    The signature of activity in general, and of stellar magnetic spots in particular, is present in every measurements, including interferometric ones. Indeed, stellar spots can be found on many stellar surfaces, their size and number varying according to their host's magnetic field and rotational velocity. To correctly determine stellar parameters, it is thus necessary to determine and extract stellar activity's signals. Interferometric observables are disturbed by activity, and this observing technique thus constitutes a good way of probing stellar surface. However, magnetic spots sometimes mimic other phenomenon, like a transiting exoplanet. In that case, the combination of several observing techniques, like photometry and interferometry, is mandatory to extract the planetary signal from the spot's one, and then characterize the exoplanet.

  19. How Many Spots Does a Cheetah Have?

    ERIC Educational Resources Information Center

    Reed, Kristine M.

    2000-01-01

    Describes first grade students' mathematical investigation of the number of spots on a cheetah. The exploration of counting and estimation strategies that grew from the investigation gives evidence that mathematicians come in all ages. (ASK)

  20. Investigations of initiation spot size effects

    SciTech Connect

    Clarke, Steven A; Akinci, Adrian A; Leichty, Gary; Schaffer, Timothy; Murphy, Michael J; Munger, Alan; Thomas, Keith A

    2010-01-01

    As explosive components become smaller, a greater understanding of the effect of initiation spot size on detonation becomes increasingly critical. A series of tests of the effect of initiation spot size will be described. A series of DOI (direct optical initiation) detonators with initiation spots sizes from {approx}50 um to 1000um have been tested to determine laser parameters for threshold firing of low density PETN pressings. Results will be compared with theoretical predictions. Outputs of the initiation source (DOI ablation) have been characterized by a suite of diagnostics including PDV and schlieren imaging. Outputs of complete detonators have been characterized using PDV, streak, and/or schlieren imaging. At present, we have not found the expected change in the threshold energy to spot size relationship for DOI type detonators found in similar earlier for projectiles, slappers and EBWs. New detonators designs (Type C) are currently being tested that will allow the determination of the threshold for spot sizes from 250 um to 105um, where we hope to see change in the threshold vs. spot size relationship. Also, one test of an extremely small diameter spot size (50um) has resulted in preliminary NoGo only results even at energy densities as much as 8 times the energy density of the threshold results presented here. This gives preliminary evidence that 50um spot may be beyond the critical initiation diameter. The constant threshold energy to spot size relationship in the data to date does however still give some insight into the initiation mechanism of DOI detonators. If the DOI initiation mechanism were a 1D mechanism similar to a slapper or a flyer impact, the expected inflection point in the graph would have been between 300um and 500um diameter spot size, within the range of the data presented here. The lack of that inflection point indicates that the DOI initiation mechanism is more likely a 2D mechanism similar to a sphere or rod projectile. We expect to

  1. White-spot disease of salmon fry

    USGS Publications Warehouse

    Mazuranich, J.J.; Nielson, W.E.

    1959-01-01

     White-spot disease, sometimes referred to as coagulated-yolk disease, has been associated with excessive mortalities occurring among the fry and early fingerling stages of the fall chinook salmon (Oncorhynchus tshawytacha) at the U.S. Fish-Cultural Stations at Carson, Cook, Underwood, and Willard, Washington. This disease of eggs and fry should not be confused with the "white-spot" infection that is caused in fingerlings by members of the protozoan genus Ichthyophthirius.

  2. HotSpot Health Physics Codes

    SciTech Connect

    Homann, S. G.

    2013-04-18

    The HotSpot Health Physics Codes were created to provide emergency response personnel and emergency planners with a fast, field-portable set of software tools for evaluating insidents involving redioactive material. The software is also used for safety-analysis of facilities handling nuclear material. HotSpot provides a fast and usually conservative means for estimation the radiation effects associated with the short-term (less than 24 hours) atmospheric release of radioactive materials.

  3. Unblinding the dark matter blind spots

    DOE PAGES

    Han, Tao; Kling, Felix; Su, Shufang; ...

    2017-02-10

    The dark matter (DM) blind spots in the Minimal Supersymmetric Standard Model (MSSM) refer to the parameter regions where the couplings of the DM particles to the $Z$-boson or the Higgs boson are almost zero, leading to vanishingly small signals for the DM direct detections. In this paper, we carry out comprehensive analyses for the DM searches under the blind-spot scenarios in MSSM. Guided by the requirement of acceptable DM relic abundance, we explore the complementary coverage for the theory parameters at the LHC, the projection for the future underground DM direct searches, and the indirect searches from the relicmore » DM annihilation into photons and neutrinos. We find that (i) the spin-independent (SI) blind spots may be rescued by the spin-dependent (SD) direct detection in the future underground experiments, and possibly by the indirect DM detections from IceCube and SuperK neutrino experiments; (ii) the detection of gamma rays from Fermi-LAT may not reach the desirable sensitivity for searching for the DM blind-spot regions; (iii) the SUSY searches at the LHC will substantially extend the discovery region for the blind-spot parameters. As a result, the dark matter blind spots thus may be unblinded with the collective efforts in future DM searches.« less

  4. Nonbright-spot AVO: Two examples

    SciTech Connect

    Ross, C.P.; Kinman, D.L.

    1995-09-01

    The use of amplitude variation with offset (AVO) attribute sections such as the product of the normal incidence trace (A) and the gradient trace (B) have been used extensively in bright spot AVO analysis and interpretation. However, while these sections have often worked well with low acoustic impedance bright spot responses, they are not reliable indicators of nonbright-spot seismic anomalies. Analyzing nonbright-spot seismic data with common AVO attribute sections will: (1) not detect the gas-charged reservoir because of near-zero acoustic impedance contrast between the sands and encasing shales, or (2) yield an incorrect (negative) AVO product if the normal incidence and gradient values are opposite in sign. The authors divide nonbright-spot AVO offset responses into two subcategories: those with phase reversals and those without. An AVO analysis procedure for these anomalies is presented through two examples. The procedure exploits the nature of the prestack response, yielding a more definitive AVO attribute section, and this technique is adaptive to both subcategories of nonbright-spot AVO responses. This technique identifies the presence of gas-charged pore fluids within the reservoir when compared to a conventionally processed, relative amplitude seismic section with characteristically low amplitude responses for near-zero acoustic impedance contrast sands.

  5. Unblinding the dark matter blind spots

    NASA Astrophysics Data System (ADS)

    Han, Tao; Kling, Felix; Su, Shufang; Wu, Yongcheng

    2017-02-01

    The dark matter (DM) blind spots in the Minimal Supersymmetric Standard Model (MSSM) refer to the parameter regions where the couplings of the DM particles to the Z-boson or the Higgs boson are almost zero, leading to vanishingly small signals for the DM direct detections. In this paper, we carry out comprehensive analyses for the DM searches under the blind-spot scenarios in MSSM. Guided by the requirement of acceptable DM relic abundance, we explore the complementary coverage for the theory parameters at the LHC, the projection for the future underground DM direct searches, and the indirect searches from the relic DM annihilation into photons and neutrinos. We find that (i) the spin-independent (SI) blind spots may be rescued by the spin-dependent (SD) direct detection in the future underground experiments, and possibly by the indirect DM detections from IceCube and SuperK neutrino experiments; (ii) the detection of gamma rays from Fermi-LAT may not reach the desirable sensitivity for searching for the DM blind-spot regions; (iii) the SUSY searches at the LHC will substantially extend the discovery region for the blind-spot parameters. The dark matter blind spots thus may be unblinded with the collective efforts in future DM searches.

  6. Canopy hot-spot as crop identifier

    SciTech Connect

    Gerstl, S.A.W.; Simmer, C.; Powers, B.J.

    1986-05-01

    Illuminating any reflective rough or structured surface by a directional light source results in an angular reflectance distribution that shows a narrow peak in the direction of retro-reflection. This is called the Heiligenschein or hot-spot of vegetation canopies and is caused by mutual shading of leaves. The angular intensity distribution of the hot-spot, its brightness and slope, are therefore indicators of the plant's geometry. We propose the use of hot-spot characteristics as crop identifiers in satellite remote sensing because the canopy hot-spot carries information about plant stand architecture that is more distinctive for different plant species than, for instance, their spectral reflectance characteristics. A simple three-dimensional Monte Carlo/ray tracing model and an analytic two-dimensional model are developed to estimate the angular distribution of the hot-spot as a function of the size of the plant leaves. The results show that the brightness-distribution and slope of the hot-spot change distinctively for different leaf sizes indicating a much more peaked maximum for the smaller leaves.

  7. Modeling deflagration waves out of hot spots

    NASA Astrophysics Data System (ADS)

    Partom, Yehuda

    2017-01-01

    It is widely accepted that shock initiation and detonation of heterogeneous explosives comes about by a two-step process known as ignition and growth. In the first step a shock sweeping through an explosive cell (control volume) creates hot spots that become ignition sites. In the second step, deflagration waves (or burn waves) propagate out of those hot spots and transform the reactant in the cell into reaction products. The macroscopic (or average) reaction rate of the reactant in the cell depends on the speed of those deflagration waves and on the average distance between neighboring hot spots. Here we simulate the propagation of deflagration waves out of hot spots on the mesoscale in axial symmetry using a 2D hydrocode, to which we add heat conduction and bulk reaction. The propagation speed of the deflagration waves may depend on both pressure and temperature. It depends on pressure for quasistatic loading near ambient temperature, and on temperature at high temperatures resulting from shock loading. From the simulation we obtain deflagration fronts emanating out of the hot spots. For 8 to 13 GPa shocks, the emanating fronts propagate as deflagration waves to consume the explosive between hot spots. For higher shock levels deflagration waves may interact with the sweeping shock to become detonation waves on the mesoscale. From the simulation results we extract average deflagration wave speeds.

  8. Dressler's Syndrome

    MedlinePlus

    ... syndrome may also be called postpericardiotomy syndrome, post-myocardial infarction syndrome and post-cardiac injury syndrome. With recent ... Dressler's syndrome. References LeWinter MM. Pericardial complications of myocardial infarction. http://www.uptodate.com/home. Accessed May 27, ...

  9. 7 CFR 28.423 - Middling Spotted Color.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 2 2014-01-01 2014-01-01 false Middling Spotted Color. 28.423 Section 28.423... REGULATIONS COTTON CLASSING, TESTING, AND STANDARDS Standards Spotted Cotton § 28.423 Middling Spotted Color. Middling Spotted Color is color which is within the range represented by a set of samples in the custody...

  10. 7 CFR 28.423 - Middling Spotted Color.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 2 2013-01-01 2013-01-01 false Middling Spotted Color. 28.423 Section 28.423... REGULATIONS COTTON CLASSING, TESTING, AND STANDARDS Standards Spotted Cotton § 28.423 Middling Spotted Color. Middling Spotted Color is color which is within the range represented by a set of samples in the custody...

  11. 7 CFR 28.423 - Middling Spotted Color.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Middling Spotted Color. 28.423 Section 28.423... REGULATIONS COTTON CLASSING, TESTING, AND STANDARDS Standards Spotted Cotton § 28.423 Middling Spotted Color. Middling Spotted Color is color which is within the range represented by a set of samples in the custody...

  12. 7 CFR 28.423 - Middling Spotted Color.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 2 2011-01-01 2011-01-01 false Middling Spotted Color. 28.423 Section 28.423... REGULATIONS COTTON CLASSING, TESTING, AND STANDARDS Standards Spotted Cotton § 28.423 Middling Spotted Color. Middling Spotted Color is color which is within the range represented by a set of samples in the custody...

  13. 7 CFR 28.423 - Middling Spotted Color.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 2 2012-01-01 2012-01-01 false Middling Spotted Color. 28.423 Section 28.423... REGULATIONS COTTON CLASSING, TESTING, AND STANDARDS Standards Spotted Cotton § 28.423 Middling Spotted Color. Middling Spotted Color is color which is within the range represented by a set of samples in the custody...

  14. HIV-related ocular microangiopathic syndrome and cognitive functioning.

    PubMed

    Geier, S A; Perro, C; Klauss, V; Naber, D; Kronawitter, U; Bogner, J R; Goebel, F D; Lund, O E; Hippius, H

    1993-03-01

    Ocular microangiopathic syndrome is found frequently in patients with AIDS or severe HIV infection. Symptoms of this microvascular syndrome can include cotton-wool spots, hemorrhages, and Roth's spots. The clinical and functional significance of HIV-related ocular microangiopathic syndrome has not been clarified as yet. The objective of this study was to evaluate a possible association between HIV-related ocular microangiopathic syndrome and cognitive functioning. Thirty-seven patients infected with HIV (24 with AIDS) underwent ophthalmological and neuropsychological examination. HIV-related ocular microangiopathic syndrome was measured by counting the number of cotton-wool spots in both eyes. Neuropsychological examination included five standardized tests, with the first three primarily measuring function of short-term memory; these tests were as follows: the Auditory-Verbal Learning Test, the Benton Test, the Stroop Colour Word Test, the Trail-Making Part B test, and the Vocabulary for Measuring Premorbid Intelligence test. HIV-related ocular microangiopathic syndrome was found in 15 patients with AIDS (62.5%), and in one patient, staged Walter Reed 5. In 10 patients, one eye was affected (mean count of cotton-wool spots 1.5). In six patients, both eyes were affected (mean count of cotton-wool spots 7.0). Univariate correlations between the number of cotton-wool spots in both eyes and test scores were as follows: Auditory-Verbal Learning Test: 0.56 (p < 0.001); Benton Test: 0.51 (p < 0.001); Stroop Colour and Word: 0.50 (p < 0.001); Trail-Making Part B: 0.15 (not significant); Vocabulary for Measuring Premorbid Intelligence: -0.05 (not significant). Multiple correlation between the test scores and the number of cotton-wool spots was 0.70 (p < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

  15. SPOTTED STAR LIGHT CURVES WITH ENHANCED PRECISION

    SciTech Connect

    Wilson, R. E.

    2012-09-15

    The nearly continuous timewise coverage of recent photometric surveys is free of the large gaps that compromise attempts to follow starspot growth and decay as well as motions, thereby giving incentive to improve computational precision for modeled spots. Due to the wide variety of star systems in the surveys, such improvement should apply to light/velocity curve models that accurately include all the main phenomena of close binaries and rotating single stars. The vector fractional area (VFA) algorithm that is introduced here represents surface elements by small sets of position vectors so as to allow accurate computation of circle-triangle overlap by spherical geometry. When computed by VFA, spots introduce essentially no noticeable scatter in light curves at the level of one part in 10,000. VFA has been put into the Wilson-Devinney light/velocity curve program and all logic and mathematics are given so as to facilitate entry into other such programs. Advantages of precise spot computation include improved statistics of spot motions and aging, reduced computation time (intrinsic precision relaxes needs for grid fineness), noise-free illustration of spot effects in figures, and help in guarding against false positives in exoplanet searches, where spots could approximately mimic transiting planets in unusual circumstances. A simple spot growth and decay template quantifies time profiles, and specifics of its utilization in differential corrections solutions are given. Computational strategies are discussed, the overall process is tested in simulations via solutions of synthetic light curve data, and essential simulation results are described. An efficient time smearing facility by Gaussian quadrature can deal with Kepler mission data that are in 30 minute time bins.

  16. Hubble Finds New Dark Spot on Neptune

    NASA Technical Reports Server (NTRS)

    1995-01-01

    NASA's Hubble Space Telescope has discovered a new great dark spot, located in the northern hemisphere of the planet Neptune. Because the planet's northern hemisphere is now tilted away from Earth, the new feature appears near the limb of the planet.

    The spot is a near mirror-image to a similar southern hemisphere dark spot that was discovered in 1989 by the Voyager 2 probe. In 1994, Hubble showed that the southern dark spot had disappeared.

    Like its predecessor, the new spot has high altitude clouds along its edge, caused by gasses that have been pushed to higher altitudes where they cool to form methane ice crystal clouds. The dark spot may be a zone of clear gas that is a window to a cloud deck lower in the atmosphere.

    Planetary scientists don t know how long lived this new feature might be. Hubble's high resolution will allow astronomers to follow the spot's evolution and other unexpected changes in Neptune's dynamic atmosphere.

    The image was taken on November 2, 1994 with Hubble's Wide Field Planetary Camera 2, when Neptune was 2.8 billion miles (4.5 billion kilometers) from Earth. Hubble can resolve features as small as 625 miles (1,000 kilometers) across in Neptune's cloud tops.

    The Wide Field/Planetary Camera 2 was developed by the Jet Propulsion Laboratory and managed by the Goddard Spaced Flight Center for NASA's Office of Space Science.

    This image and other images and data received from the Hubble Space Telescope are posted on the World Wide Web on the Space Telescope Science Institute home page at URL http://oposite.stsci.edu/pubinfo/

  17. Cushing's Syndrome

    MedlinePlus

    ... example, polycystic ovary syndrome can cause menstrual disturbances, weight gain beginning in adolescence, excess hair growth, and impaired insulin action and diabetes. Metabolic syndrome-a combination of ...

  18. Reaction of sorghum lines to zonate leaf spot and rough leaf spot

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Abundant, frequent rains, along with humid and cloudy conditions during the early part of the 2015 growing season, provided conducive conditions for an unusually severe outbreak of zonate leaf spot and rough leaf spot in a block of sorghum lines at the Texas A&M AgriLife Research Farm, Burleson Coun...

  19. Superoscillating electron wave functions with subdiffraction spots

    NASA Astrophysics Data System (ADS)

    Remez, Roei; Tsur, Yuval; Lu, Peng-Han; Tavabi, Amir H.; Dunin-Borkowski, Rafal E.; Arie, Ady

    2017-03-01

    Almost one and a half centuries ago, Abbe [Arch. Mikrosk. Anat. 9, 413 (1873), 10.1007/BF02956173] and shortly after Lord Rayleigh [Philos. Mag. Ser. 5 8, 261 (1879), 10.1080/14786447908639684] showed that, when an optical lens is illuminated by a plane wave, a diffraction-limited spot with radius 0.61 λ /sinα is obtained, where λ is the wavelength and α is the semiangle of the beam's convergence cone. However, spots with much smaller features can be obtained at the focal plane when the lens is illuminated by an appropriately structured beam. Whereas this concept is known for light beams, here, we show how to realize it for a massive-particle wave function, namely, a free electron. We experimentally demonstrate an electron central spot of radius 106 pm, which is more than two times smaller than the diffraction limit of the experimental setup used. In addition, we demonstrate that this central spot can be structured by adding orbital angular momentum to it. The resulting superoscillating vortex beam has a smaller dark core with respect to a regular vortex beam. This family of electron beams having hot spots with arbitrarily small features and tailored structures could be useful for studying electron-matter interactions with subatomic resolution.

  20. The development and validation of a fast and robust dried blood spot based lipid profiling method to study infant metabolism.

    PubMed

    Koulman, Albert; Prentice, Philippa; Wong, Max C Y; Matthews, Lee; Bond, Nicholas J; Eiden, Michael; Griffin, Julian L; Dunger, David B

    2014-01-01

    Early life exposures and metabolic programming are associated with later disease risk. In particular lipid metabolism is thought to play a key role in the development of the metabolic syndrome and insulin resistance in later life. Investigative studies of metabolic programming are limited by the ethics and practicalities of sample collection in small infants. Dried blood spots on filter paper, derived from heel pricks are considered as the most suitable option for this age group. We validated a novel lipid profiling method, based on high resolution mass spectrometry to successfully determine the lipid composition of infants using dried blood spots. The spotting and air drying of blood on paper has noticeable effects on many of the lipids, leading to lipid oxidation and hydrolysis, which demand careful interpretation of the obtained data. We compared the lipid profiles from plasma or whole blood samples and the results from dried blood spots to determine if these revealed the same inter-subject differences. The results from dried blood spots were no less reproducible than other lipid profiling methods which required comparatively larger sample volumes. Therefore, lipid profiles obtained from dried blood spots can be successfully used to monitor infancy lipid metabolism and we show significant differences in the lipid metabolism of infants at age 3 versus 12 months.

  1. Cartographic potential of SPOT image data

    NASA Technical Reports Server (NTRS)

    Welch, R.

    1985-01-01

    In late 1985, the SPOT (Systeme Probatoire d'Observation de la Terre) satellite is to be launched by the Ariane rocket from French Guiana. This satellite will have two High Resolution Visible (HRV) line array sensor systems which are capable of providing monoscopic and stereoscopic coverage of the earth. Cartographic applications are related to the recording of stereo image data and the acquisition of 20-m data in a multispectral mode. One of the objectives of this study involves a comparison of the suitability of SPOT and TM image data for mapping urban land use/cover. Another objective is concerned with a preliminary assessment of the potential of SPOT image data for map revision when merged with conventional map sheets converted to raster formats.

  2. On turbulent spots in plane Poiseuille flow

    NASA Technical Reports Server (NTRS)

    Henningson, Dan S.; Kim, John

    1991-01-01

    Turbulence characteristics inside a turbulent spot in plane Poiseuille flow are investigated by analyzing a database obtained from a direct numerical simulation. The spot is found to consist of two distinct regions - a turbulent area and a wave area. The flow inside the turbulent area has a strong resemblance to that found in the fully developed turbulent channel. Suitably defined mean and r.m.s. fluctuations as well as the internal shear-layer structures are found to be similar to the turbulent counterpart. In the wave area the inflexional mean spanwise profiles cause a rapid growth of oblique waves, which break down to turbulence. The breakdown process of the oblique waves is reminiscent of the secondary instability observed during transition to turbulence in channel and boundary-layer flows. Other detailed characteristics associated with the Poiseuille spot are presented and are compared with experimental results.

  3. Fatal Spotted Fever Rickettsiosis, Minas Gerais, Brazil

    PubMed Central

    Dumler, J. Stephen; Mafra, Cláudio Lísias; Calic, Simone Berger; Chamone, Chequer Buffe; Filho, Gracco Cesarino; Olano, Juan Pablo; Walker, David H.

    2003-01-01

    The emergence and reemergence of a serious infectious disease are often associated with a high case-fatality rate because of misdiagnosis and inappropriate or delayed treatment. The current reemergence of spotted fever rickettsiosis caused by Rickettsia rickettsii in Brazil has resulted in a high proportion of fatal cases. We describe two familial clusters of Brazilian spotted fever in the state of Minas Gerais, involving six children 9 months to 15 years of age; five died. Immunohistochemical investigation of tissues obtained at necropsy of a child in each location, Novo Cruzeiro and Coronel Fabriciano municipalities, established the diagnosis by demonstration of disseminated endothelial infection with spotted fever group rickettsiae. The diagnosis in the two fatal cases from Coronel Fabriciano and the surviving patient from Novo Cruzeiro was further supported by immunofluorescence serologic tests. PMID:14718082

  4. [Rocky Mountain spotted fever in Brazil].

    PubMed

    del Sá DelFiol, Fernando; Junqueira, Fábio Miranda; da Rocha, Maria Carolina Pereira; de Toledo, Maria Inês; Filho, Silvio Barberato

    2010-06-01

    Although the number of confirmed cases of spotted fever has been declining in Brazil since 2005, the mortality rate (20% to 30%) is still high in comparison to other countries. This high mortality rate is closely related to the difficulty in making the diagnosis and starting the correct treatment. Only two groups of antibiotics have proven clinical effectiveness against spotted fever: chloramphenicol and tetracyclines. Until recently, the use of tetracyclines was restricted to adults because of the associated bone and tooth changes in children. Recently, however, the American Academy of Pediatrics and various researchers have recommended the use of doxycycline in children. In more severe cases, chloramphenicol injections are often preferred in Brazil because of the lack of experience with injectable tetracycline. Since early diagnosis and the adequate drug treatment are key to a good prognosis, health care professionals must be better prepared to recognize and treat spotted fever.

  5. On turbulent spots in plane Poiseuille flow

    NASA Technical Reports Server (NTRS)

    Henningson, Dan S.; Kim, John

    1992-01-01

    Turbulence characteristics inside a turbulent spot in plume Poiseuille flow are investigated by analyzing a data base obtained from a direct numerical simulation. The spot is found to consist of two distinct regions - a turbulent area and a wave area. The flow inside the turbulent area has a strong resemblance to that found in the fully developed turbulent channel. Suitably defined mean and rms fluctuations as well as the internal shear layer structures are found to be similar to the turbulent counterpart. In the wave area, the inflexional mean spanwise profiles cause a rapid growth of oblique waves, which break down to turbulence. The breakdown process of the oblique waves is reminiscent of the secondary instability observed during transition to turbulence in channel and boundary layer flows. Other detailed characteristics associated with the Poiseuille spot are presented and are compared with experimental results.

  6. Spots on AG Virginis - Paradigm or panacea?

    NASA Astrophysics Data System (ADS)

    Bell, S. A.; Rainger, P. P.; Hilditch, R. W.

    1990-12-01

    New photometric and spectroscopic observations of the eclipsing binary AG Vir are presented. Medium-resolution spectroscopy has allowed the measurement of velocities for the secondary component for the first time. The V light curve shows many of the features seen in previous studies of this system. A full analysis of the spectroscopic and photometric data has been made which suggests that the system is either in a marginal state of conatact or a deep-contact configuration depending on the type of spot model invoked. AG Vir constitutes an excellent example of the expected manifestations of spot activity on a light curve. It also demonstrates the ease with which the spot phenomenon can be invoked to explain the appearance of a light curve and to provide conflicting results. This study shows the necessity of a more thorough investigation of this system using Doppler-imaging techniques and simultaneous infrared and optical photometry.

  7. Development of hot spot fixer (HSF)

    NASA Astrophysics Data System (ADS)

    Kotani, Toshiya; Kyoh, Suigen; Kobayashi, Sachiko; Inazu, Takatoshi; Ikeuchi, Atsuhiko; Urakawa, Yukihiro; Inoue, Soichi; Morita, Etsuya; Klaver, Simon; Horiuchi, Takumi; Peeters, Johan; Kuramoto, Satoshi

    2006-03-01

    A new design for manufacturability (DfM) scheme with a lithography compliance check (LCC) and hot spot fixing (HSF) flow has been developed to guarantee design compliance for OPC and RET by combining lithography simulator, hot spot detector and layout modification tool. Hot spots highlighted by the LCC flow are removed by the HSF flow following modification rule consists of "Line-Sizing" (LS) and "Space-Sizing (SS)" that are resize value of line-width and space-width for the original pattern. In order to meet layout modification requirements at the pre- and post- tape out (T.O.) stages, the priorities individually set for the modification rules and the design rules, which provides flexibly to achieve the modification scheme desirable at each stage. For handling large data at a fast speed, Layout Analyzer (LA) and Layout Optimizer (LO) engines were combined with the HSF flow. LA is used to reconstruct the original hierarchy structure, clips off small parts of the layout that include hot spots from the original layout and sends those to LO in order to reduce the computational time and resource. LO optimizes the clipped off layout following the prioritized modification- and design-rules. The new DfM scheme was found to be quite effective for hot spot cleaning for 65nm node and beyond, since it was demonstrated that the HSF flow improved the lithography margin for the metal layer of 65nm node full-chip data by reducing number of hot spots to below 0.1% of original within about 12 hours, using 1CPU of commercially available workstation.

  8. Statistical Hot Spot Model for Explosive Detonation

    SciTech Connect

    Nichols, III, A L

    2005-07-14

    The Non-local Thermodynamic Equilibrium Statistical Hot Spot Model (NLTE SHS), a new model for explosive detonation, is described. In this model, the formation, ignition, propagation, and extinction of hot spots is explicitly modeled. The equation of state of the explosive mixture is treated with a non-local equilibrium thermodynamic assumption. A methodology for developing the parameters for the model is discussed, and applied to the detonation velocity diameter effect. Examination of these results indicates where future improvements to the model can be made.

  9. Statistical Hot Spot Model for Explosive Detonation

    SciTech Connect

    Nichols III, A L

    2004-05-10

    The Non-local Thermodynamic Equilibrium Statistical Hot Spot Model (NLTE SHS), a new model for explosive detonation, is described. In this model, the formation, ignition, propagation, and extinction of hot spots is explicitly modeled. The equation of state of the explosive mixture is treated with a nonlocal equilibrium thermodynamic assumption. A methodology for developing the parameters for the model is discussed, and applied to the detonation velocity diameter effect. Examination of these results indicates where future improvements to the model can be made.

  10. The simple spot-welding apparatus

    NASA Astrophysics Data System (ADS)

    Hiraoka, Takeshi

    1998-07-01

    A simple spot-welding apparatus has been developed for the fabrication of stable electrodes used in the measurement of transport properties in condensed matter physics. The apparatus is especially suited for the use of brittle and small sized samples of rare-earth intermetallic compounds (REICs). The spot welding is made by a sharp pulse (150 A and several μs), generated by the precise time generation IC in a capacitor discharge circuit, to avoid breaking the sample. Stable electrodes of six 15 μ Au wires can be made on a REIC sample with length less than 1 mm.

  11. Jorge Rivera congratulated for spotting misplaced pin

    NASA Technical Reports Server (NTRS)

    2000-01-01

    Jorge Rivera (with microphone) shares his thoughts after receiving congratulations for his observatory powers from NASA Administrator Dan Goldin (right). A NASA external tank mechanical engineer, Rivera is the one who spotted the misplaced lock pin on Shuttle Discovery Oct. 10, shortly before the intended launch of mission STS-92, causing a scrub for safety reasons.

  12. Jorge Rivera congratulated for spotting misplaced pin

    NASA Technical Reports Server (NTRS)

    2000-01-01

    Jorge Rivera (center) receives a plaque and congratulations for his observatory powers from Shuttle Launch Director Mike Leinbach (right). A NASA external tank mechanical engineer, Rivera is the one who spotted the misplaced lock pin on Shuttle Discovery Oct. 10, shortly before the intended launch of mission STS-92, causing a scrub for safety reasons.

  13. The sweet spots in human communication.

    PubMed

    Salem, Philip

    2011-07-01

    In baseball, the sweet spot is a special place on a bat where the batter can hit the ball with the most power. It is the place where the performances of the batter and pitcher collide with maximum effect. It is the place where the dynamic tension between opponents leads to transformation. The dynamic tension in all living systems is between similarity and difference. Chaos and complexity scholars recognized this tension as amounts of information. When the amounts of information were high, but not too high, the system moved to the edge of chaos, to the complexity regime, to strange attractors, or to chaos, depending on the model. The sweet spot is that range of relative variety, just the proper mix of similarity and difference, leading to transformation. This essay contains a model of human communication as an emergent social process with its own sweet spots. The essay also includes a description of current literature highlighting tensions between similarity and difference, and there is an exploration of the potential to move from one basin of attraction to another. The primary constraints on finding communication sweet spots are paradigmatic - adopting a process orientation, discovering the proper parameters, bracketing sequences to define initial conditions, and understanding the strengths and weaknesses of various modeling techniques.

  14. 9 CFR 149.4 - Spot audit.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Spot audit. 149.4 Section 149.4 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE... results as a result of testing of certified swine from that site at the slaughter facility. (b) All...

  15. 9 CFR 149.4 - Spot audit.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Spot audit. 149.4 Section 149.4 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF AGRICULTURE... results as a result of testing of certified swine from that site at the slaughter facility. (b) All...

  16. Arc spot welding technique for underwater use

    SciTech Connect

    Koga, H.; Ide, Y.; Ogawa, Y.

    1995-12-31

    An arc spot welding equipment with special local cavity shroud was developed for underwater salvaging activity. Arc spot welding for lapped plates is an effective method to recover defects. This method in surface is so simple to use widely in the field of railways and chemical plants manufacturing. But there is some problems on the reliability of joint strength and bead shapes. A special arc spot nozzle to improve welding quality was developed. A small outlet of air jet at the bottom of the nozzle was created to maintain the swirl flow of shielding gas and certain rejection of excessive molten metal. This nozzle covers the welding part completely, then it also works as a local cavity shroud under water. This paper describes the design and function of the nozzle for CO{sub 2} arc spot welding system. A programmable controller manages the welding sequence of shielding gas flow, air jet flow, and arcing time. This welding gun is operated manually, but the operation is only to press the gun on the weld point. After that welding will proceed automatically, and arcing time is about three seconds. Whole time for welding which includes pre and post gas flow time is less than ten seconds for surface use, it is required some more additional pre drying process of welding point for underwater use to guarantee the high quality welding results. Fundamental analysis of welding conditions and the effects of air jet were considered.

  17. Turbulent Spots Inside the Turbulent Boundary Layer

    NASA Astrophysics Data System (ADS)

    Skarda, Jinhie; Wu, Xiaohua; Moin, Parviz; Lozano-Duran, Adrian; Wallace, James; Hickey, Jean-Pierre

    2016-11-01

    We present evidence that the buffer region of the canonical turbulent boundary layer is populated by locally generated turbulent spots, which cause strong indentations on the near-wall low-momentum streaks. This evidence is obtained from a spatially-developing direct numerical simulation carrying the inlet Blasius boundary layer through a bypass transition to the turbulent boundary layer state over a moderate Reynolds number range. The turbulent spots are structurally analogous to their transitional counter-parts but without any direct causality connection. High-pass filtered time-history records are used to calculate the period of turbulent spot detection and this period is compared to the boundary layer bursting period reported in hot-wire experiments. The sensitivity of the results to parameters such as the high pass filter frequency and the amplitude discriminator level is examined. The characteristics of these turbulent spots are also quantified using a spatial connectivity based conditional sampling technique. This evidence seems to be at odds with the notion that the buffer region is dominated by quasi-streamwise vortices, and contributes to the potential unification of the studies on near-wall turbulent boundary layer dynamics.

  18. THE SPACE PUBLIC OUTREACH TEAM (SPOT)

    NASA Astrophysics Data System (ADS)

    Williamson, Kathryn; National Radio Astronomy Observatory; Montana Space Grant Consortium; West Virginia Space Grant Consortium; NASA Independent Verification and Validation Center

    2014-01-01

    The Space Public Outreach Team (SPOT) has shown over 17 years of success in bringing astronomy and space science-themed presentations to approximately 10,000 students per year in Montana, and the program is now being piloted in West Virginia through a joint partnership between the National Radio Astronomy Observatory (NRAO), the West Virginia Space Grant Consortium, and NASA Independent Verification and Validation Center. SPOT recruits and trains undergraduate presenters from all over the state to learn interactive slide shows that highlight the state’s on-going and world-class space science research. Presenters then travel to K-12 schools to deliver these presentations and provide teachers additional supplemental information for when the SPOT team leaves. As a large-scale, low-cost, and sustainable program being implemented in both Montana and West Virginia, SPOT has the potential to become a nation-wide effort that institutions in other states can model to increase their education and public outreach presence.

  19. Easy Demonstration of the Poisson Spot

    ERIC Educational Resources Information Center

    Gluck, Paul

    2010-01-01

    Many physics teachers have a set of slides of single, double and multiple slits to show their students the phenomena of interference and diffraction. Thomas Young's historic experiments with double slits were indeed a milestone in proving the wave nature of light. But another experiment, namely the Poisson spot, was also important historically and…

  20. Expert Blind Spot among Preservice Teachers

    ERIC Educational Resources Information Center

    Nathan, Mitchell J.; Petrosino, Anthony

    2003-01-01

    This study (N = 48) examined the relationship between preservice secondary teachers' subject-matter expertise in mathematics and their judgments of students' algebra problem-solving difficulty. As predicted by the "expert blind spot" hypothesis, participants with more advanced mathematics education, regardless of their program affiliation or…

  1. Triggered tremor sweet spots in Alaska

    USGS Publications Warehouse

    Gomberg, Joan; Prejean, Stephanie

    2013-01-01

    To better understand what controls fault slip along plate boundaries, we have exploited the abundance of seismic and geodetic data available from the richly varied tectonic environments composing Alaska. A search for tremor triggered by 11 large earthquakes throughout all of seismically monitored Alaska reveals two tremor “sweet spots”—regions where large-amplitude seismic waves repeatedly triggered tremor between 2006 and 2012. The two sweet spots locate in very different tectonic environments—one just trenchward and between the Aleutian islands of Unalaska and Akutan and the other in central mainland Alaska. The Unalaska/Akutan spot corroborates previous evidence that the region is ripe for tremor, perhaps because it is located where plate-interface frictional properties transition between stick-slip and stably sliding in both the dip direction and laterally. The mainland sweet spot coincides with a region of complex and uncertain plate interactions, and where no slow slip events or major crustal faults have been noted previously. Analyses showed that larger triggering wave amplitudes, and perhaps lower frequencies (<~0.03 Hz), may enhance the probability of triggering tremor. However, neither the maximum amplitude in the time domain or in a particular frequency band, nor the geometric relationship of the wavefield to the tremor source faults alone ensures a high probability of triggering. Triggered tremor at the two sweet spots also does not occur during slow slip events visually detectable in GPS data, although slow slip below the detection threshold may have facilitated tremor triggering.

  2. Hot-spot tectonics on Io

    NASA Technical Reports Server (NTRS)

    Mcewen, A. S.

    1985-01-01

    The thesis is that extensional tectonics and low-angle detachment faults probably occur on Io in association with the hot spots. These processes may occur on a much shorter timescale on Ion than on Earth, so that Io could be a natural laboratory for the study of thermotectonics. Furthermore, studies of heat and detachment in crustal extension on Earth and the other terresrial planets (especially Venus and Mars) may provide analogs to processes on Io. The geology of Io is dominated by volcanism and hot spots, most likely the result of tidal heating. Hot spots cover 1 to 2% of Io's surface, radiating at temperatures typically from 200 to 400 K, and occasionally up to 700K. Heat loss from the largest hot spots on Io, such as Loki Patera, is about 300 times the heat loss from Yellowstone, so a tremendous quantity of energy is available for volcanic and tectonic work. Active volcanism on Io results in a resurfacing rate as high as 10 cm per year, yet many structural features are apparent on the surface. Therefore, the tectonics must be highly active.

  3. The Power Company Hot-Spot

    NASA Astrophysics Data System (ADS)

    Barresi, Joseph F.

    1984-03-01

    Just as your companies screen and monitor the market place to determine demand and the public's perception of product need, the power company is constantly faced with studying and forecasting the potential for load growth within it's franchised areas. This study and it's resultant forecast for growth has placed the power industry in a hot spot.

  4. Digital Sun Sensor Multi-Spot Operation

    PubMed Central

    Rufino, Giancarlo; Grassi, Michele

    2012-01-01

    The operation and test of a multi-spot digital sun sensor for precise sun-line determination is described. The image forming system consists of an opaque mask with multiple pinhole apertures producing multiple, simultaneous, spot-like images of the sun on the focal plane. The sun-line precision can be improved by averaging multiple simultaneous measures. Nevertheless, the sensor operation on a wide field of view requires acquiring and processing images in which the number of sun spots and the related intensity level are largely variable. To this end, a reliable and robust image acquisition procedure based on a variable shutter time has been considered as well as a calibration function exploiting also the knowledge of the sun-spot array size. Main focus of the present paper is the experimental validation of the wide field of view operation of the sensor by using a sensor prototype and a laboratory test facility. Results demonstrate that it is possible to keep high measurement precision also for large off-boresight angles. PMID:23443388

  5. Spotting Junk Science A Classroom Exercise

    NASA Astrophysics Data System (ADS)

    Houser, Brian

    2006-12-01

    This presentation is on an exercise in teaching non-science students to spot junk science and to be better consumers of scientific information. It is based upon critical analysis of the website advocating the theory of the solid surface of the sun. I will present examples of student work and solicit comments on how to improve the exercise.

  6. Two New Hot Spots on Io

    NASA Technical Reports Server (NTRS)

    1997-01-01

    The Near-Infrared Mapping Spectrometer (NIMS) on Galileo obtained this image of half of Io's disk in darkness on September 19, 1997. This image, at 5 microns, shows several hot spots on Io, which are volcanic regions of enhanced thermal emission. The area shown is part of the leading hemisphere of Io.

    Two new hot spots are shown and indicated in the image (New, and Shamshu). Neither of these hot spots were seen by NIMS or the Solid State Imaging Experiment, (SSI) prior to this observation, becoming only recently active. Several other previously known hot spots are labelled in the image. Galileo was at a distance of 342,000 km from Io when this observation was made.

    The Jet Propulsion Laboratory, Pasadena, CA manages the mission for NASA's Office of Space Science, Washington, DC.

    The Jet Propulsion Laboratory, Pasadena, CA manages the mission for NASA's Office of Space Science, Washington, DC.

    This image and other images and data received from Galileo are posted on the World Wide Web, on the Galileo mission home page at URL http://galileo.jpl.nasa.gov.

  7. Rocky Mountain spotted fever in dogs, Brazil.

    PubMed

    Labruna, Marcelo B; Kamakura, Orson; Moraes-Filho, Jonas; Horta, Mauricio C; Pacheco, Richard C

    2009-03-01

    Clinical illness caused by Rickettsia rickettsii in dogs has been reported solely in the United States. We report 2 natural clinical cases of Rocky Mountain spotted fever in dogs in Brazil. Each case was confirmed by seroconversion and molecular analysis and resolved after doxycycline therapy.

  8. Dramatic Change in Jupiter's Great Red Spot

    NASA Technical Reports Server (NTRS)

    Simon, A. A.; Wong, M. H.; Rogers, J. H.; Orton, G. S.; de Pater, I.; Asay-Davis, X.; Carlson, R. W.; Marcus, P. S.

    2015-01-01

    Jupiter's Great Red Spot (GRS) is one of its most distinct and enduring features, having been continuously observed since the 1800's. It currently spans the smallest latitude and longitude size ever recorded. Here we show analyses of 2014 Hubble spectral imaging data to study the color, structure and internal dynamics of this long-live storm.

  9. Mutation hot spots in mammalian mitochondrial DNA.

    PubMed

    Galtier, Nicolas; Enard, David; Radondy, Yoan; Bazin, Eric; Belkhir, Khalid

    2006-02-01

    Animal mitochondrial DNA is characterized by a remarkably high level of within-species homoplasy, that is, phylogenetic incongruence between sites of the molecule. Several investigators have invoked recombination to explain it, challenging the dogma of maternal, clonal mitochondrial inheritance in animals. Alternatively, a high level of homoplasy could be explained by the existence of mutation hot spots. By using an exhaustive mammalian data set, we test the hot spot hypothesis by comparing patterns of site-specific polymorphism and divergence in several groups of closely related species, including hominids. We detect significant co-occurrence of synonymous polymorphisms among closely related species in various mammalian groups, and a correlation between the site-specific levels of variability within humans (on one hand) and between Hominoidea species (on the other hand), indicating that mutation hot spots actually exist in mammalian mitochondrial coding regions. The whole data, however, cannot be explained by a simple mutation hot spots model. Rather, we show that the site-specific mutation rate quickly varies in time, so that the same sites are not hypermutable in distinct lineages. This study provides a plausible mutation model that potentially accounts for the peculiar distribution of mitochondrial sequence variation in mammals without the need for invoking recombination. It also gives hints about the proximal causes of mitochondrial site-specific hypermutability in humans.

  10. Measuring microfocal spots using digital radiography

    SciTech Connect

    Fry, David A; Ewert, Uwe

    2009-01-01

    Measurement of microfocus spot size can be important for several reasons: (1) Quality assurance during manufacture of microfocus tubes; (2) Tracking performance and stability of microfocus tubes; (3) Determining magnification is especially important for digital radiography where the native spatial resolution of the digital system is not adequate for the application; and (4) Knowledge of unsharpness from the focal spot alone. The European Standard EN 12543-5 is based on a simple geometrical method of calculating focal spot size from unsharpness of high magnification film radiographs. The following equations are used for the focal spot size measurement: By similar triangles the following equations are presupposed: f/a = U/b and M = (a+b)/a. These equations can be combined to yield the well known expression: U = f(M - 1). Solving for f, f = U/(M-1). Therefore, the focal spot size, f, can be calculated by measuring the radiographic unsharpness and magnification of a known object. This is the basis for these tests. The European standard actually uses one-half of the unsharpness (which are then added together) from both sides of the object to avoid additional unsharpness contributions due to edge transmission unsharpness of the round test object (the outside of the object is measured). So the equation becomes f = (1/2 U{sub 1} + 1/2 U{sub 2})/(M-1). In practice 1/2 U is measured from the 50% to the 90% signal points on the transition profile from ''black'' to ''white,'' (positive image) or attenuated to unattenuated portion of the image. The 50% to 90% points are chosen as a best fit to an assumed Gaussian radiation distribution from the focal spot and to avoid edge transmission effects. 1/2 U{sub 1} + 1/2 U{sub 2} corresponds about to the full width at half height of a Gaussian focal spot. A highly absorbing material (Tungsten, Tungsten Alloy, or Platinum) is used for the object. Either wires or a sphere are used as the object to eliminate alignment issues. One

  11. The Little Red Spot: Closest View Yet

    NASA Technical Reports Server (NTRS)

    2007-01-01

    This is a mosaic of three New Horizons images of Jupiter's Little Red Spot, taken with the spacecraft's Long Range Reconnaissance Imager (LORRI) camera at 17:41 Universal Time on February 26 from a range of 3.5 million kilometers (2.1 million miles). The image scale is 17 kilometers (11 miles) per pixel, and the area covered measures 33,000 kilometers (20,000 miles) from top to bottom, two and one-half times the diameter of Earth.

    The Little Red Spot, a smaller cousin of the famous Great Red Spot, formed in the past decade from the merger of three smaller Jovian storms, and is now the second-largest storm on Jupiter. About a year ago its color, formerly white, changed to a reddish shade similar to the Great Red Spot, perhaps because it is now powerful enough to dredge up reddish material from deeper inside Jupiter. These are the most detailed images ever taken of the Little Red Spot since its formation, and will be combined with even sharper images taken by New Horizons 10 hours later to map circulation patterns around and within the storm.

    LORRI took the images as the Sun was about to set on the Little Red Spot. The LORRI camera was designed to look at Pluto, where sunlight is much fainter than it is at Jupiter, so the images would have been overexposed if LORRI had looked at the storm when it was illuminated by the noonday Sun. The dim evening illumination helped the LORRI camera obtain well-exposed images. The New Horizons team used predictions made by amateur astronomers in 2006, based on their observations of the motion of the Little Red Spot with backyard telescopes, to help them accurately point LORRI at the storm.

    These are among a handful of Jupiter system images already returned by New Horizons during its close approach to Jupiter. Most of the data being gathered by the spacecraft are stored onboard and will be downlinked to Earth during March and April 2007.

  12. Hyperspectral analysis of columbia spotted frog habitat

    USGS Publications Warehouse

    Shive, J.P.; Pilliod, D.S.; Peterson, C.R.

    2010-01-01

    Wildlife managers increasingly are using remotely sensed imagery to improve habitat delineations and sampling strategies. Advances in remote sensing technology, such as hyperspectral imagery, provide more information than previously was available with multispectral sensors. We evaluated accuracy of high-resolution hyperspectral image classifications to identify wetlands and wetland habitat features important for Columbia spotted frogs (Rana luteiventris) and compared the results to multispectral image classification and United States Geological Survey topographic maps. The study area spanned 3 lake basins in the Salmon River Mountains, Idaho, USA. Hyperspectral data were collected with an airborne sensor on 30 June 2002 and on 8 July 2006. A 12-year comprehensive ground survey of the study area for Columbia spotted frog reproduction served as validation for image classifications. Hyperspectral image classification accuracy of wetlands was high, with a producer's accuracy of 96 (44 wetlands) correctly classified with the 2002 data and 89 (41 wetlands) correctly classified with the 2006 data. We applied habitat-based rules to delineate breeding habitat from other wetlands, and successfully predicted 74 (14 wetlands) of known breeding wetlands for the Columbia spotted frog. Emergent sedge microhabitat classification showed promise for directly predicting Columbia spotted frog egg mass locations within a wetland by correctly identifying 72 (23 of 32) of known locations. Our study indicates hyperspectral imagery can be an effective tool for mapping spotted frog breeding habitat in the selected mountain basins. We conclude that this technique has potential for improving site selection for inventory and monitoring programs conducted across similar wetland habitat and can be a useful tool for delineating wildlife habitats. ?? 2010 The Wildlife Society.

  13. Revised Thorium Abundances for Lunar Red Spots

    NASA Technical Reports Server (NTRS)

    Hagerty, J. J.; Lawrence, D. J.; Elphic, R. C.; Feldman, W. C.; Vaniman, D. T.; Hawke, B. R.

    2005-01-01

    Lunar red spots are features on the nearside of the Moon that are characterized by high albedo and by a strong absorption in the ultraviolet. These red spots include the Gruithuisen domes, the Mairan domes, Hansteen Alpha, the southern portion of Montes Riphaeus, Darney Chi and Tau, Helmet, and an area near the Lassell crater. It has been suggested that many of the red spots are extrusive, nonmare, volcanic features that could be composed of an evolved lithlogy enriched in thorium. In fact, Hawke et al. used morphological characteristics to show that Hansteen Alpha is a nonmare volcanic construct. However, because the apparent Th abundances (6 - 7 ppm) were lower than that expected for evolved rock types, Hawke et al. concluded that Hansteen Alpha was composed of an unknown rock type. Subsequent studies by Lawrence et al. used improved knowledge of the Th spatial distribution for small area features on the lunar surface to revisit the interpretation of Th abundances at the Hansteen Alpha red spot. As part of their study, Lawrence et al. used a forward modeling technique to show that the Th abundance at Hansteen Alpha is not 6 ppm, but is more likely closer to 25 ppm, a value consistent with evolved lithologies. This positive correlation between the morphology and composition of Hansteen Alpha provides support for the presence of evolved lithologies on the lunar surface. It is possible, however, that Hansteen Alpha represents an isolated occurrence of non-mare volcanism. That is why we have chosen to use the forward modeling technique of Lawrence et al. to investigate the Th abundances at other lunar red spots, starting with the Gruithuisen domes. Additional information is included in the original extended abstract.

  14. Tests to Spot 'Preemie' Birth Risk Ineffective in First Pregnancies

    MedlinePlus

    ... news/fullstory_164081.html Tests to Spot 'Preemie' Birth Risk Ineffective in First Pregnancies Finding suggests other ... HealthDay News) -- Two tests that screen for preterm birth risk rarely spot trouble in first-time pregnancies, ...

  15. SPOT: How good for geology? A comparison with LANDSAT MSS

    NASA Astrophysics Data System (ADS)

    Sesoeren, A.

    1986-12-01

    Geological interpretation possibilities of SPOT MSS and LANDSAT MSS positive prints enlarged to the same scale were compared, using as a test area part of the Jebel Amour (Algeria). The SPOT imagery offers many advantages, filling the gap between remote sensing from space and aerial photography. The best results by visual interpretation are obtained in combining SPOT for the required details with LANDSAT for the synoptic veiw. Further improvements are expected from the use of SPOT stereo-pairs.

  16. Vibrational modes identify soft spots in a sheared disordered packing.

    PubMed

    Manning, M L; Liu, A J

    2011-09-02

    We analyze low-frequency vibrational modes in a two-dimensional, zero-temperature, quasistatically sheared model glass to identify a population of structural "soft spots" where particle rearrangements are initiated. The population of spots evolves slowly compared to the interval between particle rearrangements, and the soft spots are structurally different from the rest of the system. Our results suggest that disordered solids flow via localized rearrangements that tend to occur at soft spots, which are analogous to dislocations in crystalline solids.

  17. Turbulent Region Near Jupiter's Great Red Spot

    NASA Technical Reports Server (NTRS)

    1997-01-01

    True and false color mosaics of the turbulent region west of Jupiter's Great Red Spot. The Great Red Spot is on the planetary limb on the right hand side of each mosaic. The region west (left) of the Great Red Spot is characterized by large, turbulent structures that rapidly change in appearance. The turbulence results from the collision of a westward jet that is deflected northward by the Great Red Spot into a higher latitude eastward jet. The large eddies nearest to the Great Red Spot are bright, suggesting that convection and cloud formation are active there.

    The top mosaic combines the violet (410 nanometers) and near infrared continuum (756 nanometers) filter images to create a mosaic similar to how Jupiter would appear to human eyes. Differences in coloration are due to the composition and abundance of trace chemicals in Jupiter's atmosphere. The lower mosaic uses the Galileo imaging camera's three near-infrared (invisible) wavelengths (756 nanometers, 727 nanometers, and 889 nanometers displayed in red, green, and blue) to show variations in cloud height and thickness. Light blue clouds are high and thin, reddish clouds are deep, and white clouds are high and thick. Purple most likely represents a high haze overlying a clear deep atmosphere. Galileo is the first spacecraft to distinguish cloud layers on Jupiter.

    The mosaic is centered at 16.5 degrees south planetocentric latitude and 85 degrees west longitude. The north-south dimension of the Great Red Spot is approximately 11,000 kilometers. The smallest resolved features are tens of kilometers in size. North is at the top of the picture. The images used were taken on June 26, 1997 at a range of 1.2 million kilometers (1.05 million miles) by the Solid State Imaging (SSI) system on NASA's Galileo spacecraft.

    The Jet Propulsion Laboratory, Pasadena, CA manages the Galileo mission for NASA's Office of Space Science, Washington, DC. JPL is an operating division of California Institute of Technology

  18. 7 CFR 28.424 - Strict Low Middling Spotted Color.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 2 2011-01-01 2011-01-01 false Strict Low Middling Spotted Color. 28.424 Section 28.424 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Spotted Color. Strict Low Middling Spotted Color is color which is within the range represented by a...

  19. 7 CFR 28.426 - Strict Good Ordinary Spotted Color.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Strict Good Ordinary Spotted Color. 28.426 Section 28.426 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Spotted Color. Strict Good Ordinary Spotted Color is color which is within the range represented by a...

  20. 7 CFR 28.426 - Strict Good Ordinary Spotted Color.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 2 2012-01-01 2012-01-01 false Strict Good Ordinary Spotted Color. 28.426 Section 28.426 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Spotted Color. Strict Good Ordinary Spotted Color is color which is within the range represented by a...

  1. 7 CFR 28.426 - Strict Good Ordinary Spotted Color.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 2 2014-01-01 2014-01-01 false Strict Good Ordinary Spotted Color. 28.426 Section 28.426 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Spotted Color. Strict Good Ordinary Spotted Color is color which is within the range represented by a...

  2. 7 CFR 28.424 - Strict Low Middling Spotted Color.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Strict Low Middling Spotted Color. 28.424 Section 28.424 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Spotted Color. Strict Low Middling Spotted Color is color which is within the range represented by a...

  3. 7 CFR 28.421 - Good Middling Spotted Color.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 2 2013-01-01 2013-01-01 false Good Middling Spotted Color. 28.421 Section 28.421 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Color. Good Middling Spotted Color is color which is better than Strict Middling Spotted Color....

  4. 7 CFR 28.424 - Strict Low Middling Spotted Color.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 2 2013-01-01 2013-01-01 false Strict Low Middling Spotted Color. 28.424 Section 28.424 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Spotted Color. Strict Low Middling Spotted Color is color which is within the range represented by a...

  5. 7 CFR 28.421 - Good Middling Spotted Color.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 2 2012-01-01 2012-01-01 false Good Middling Spotted Color. 28.421 Section 28.421 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Color. Good Middling Spotted Color is color which is better than Strict Middling Spotted Color....

  6. 7 CFR 28.426 - Strict Good Ordinary Spotted Color.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 2 2013-01-01 2013-01-01 false Strict Good Ordinary Spotted Color. 28.426 Section 28.426 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Spotted Color. Strict Good Ordinary Spotted Color is color which is within the range represented by a...

  7. 7 CFR 28.421 - Good Middling Spotted Color.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 2 2014-01-01 2014-01-01 false Good Middling Spotted Color. 28.421 Section 28.421 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Color. Good Middling Spotted Color is color which is better than Strict Middling Spotted Color....

  8. 7 CFR 28.424 - Strict Low Middling Spotted Color.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 2 2014-01-01 2014-01-01 false Strict Low Middling Spotted Color. 28.424 Section 28.424 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Spotted Color. Strict Low Middling Spotted Color is color which is within the range represented by a...

  9. 7 CFR 28.424 - Strict Low Middling Spotted Color.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 2 2012-01-01 2012-01-01 false Strict Low Middling Spotted Color. 28.424 Section 28.424 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Spotted Color. Strict Low Middling Spotted Color is color which is within the range represented by a...

  10. 7 CFR 28.421 - Good Middling Spotted Color.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 2 2011-01-01 2011-01-01 false Good Middling Spotted Color. 28.421 Section 28.421 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Color. Good Middling Spotted Color is color which is better than Strict Middling Spotted Color....

  11. 7 CFR 28.426 - Strict Good Ordinary Spotted Color.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 2 2011-01-01 2011-01-01 false Strict Good Ordinary Spotted Color. 28.426 Section 28.426 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Spotted Color. Strict Good Ordinary Spotted Color is color which is within the range represented by a...

  12. 7 CFR 28.421 - Good Middling Spotted Color.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Good Middling Spotted Color. 28.421 Section 28.421 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Color. Good Middling Spotted Color is color which is better than Strict Middling Spotted Color....

  13. Spot-Welding Gun With Pivoting Twin-Collet Assembly

    NASA Technical Reports Server (NTRS)

    Nguyen, Francis; Simpson, Gareth; Hoult, William S.

    1996-01-01

    Modified spot-welding gun includes pivoting twin-collet assembly that holds two spot-welding electrodes. Designed to weld highly conductive (30 percent gold) brazing-alloy foils to thin nickel alloy workpieces; also suitable for other spot-welding applications compatible with two-electrode configuration.

  14. Spot-Welding Gun Is Easy To Use

    NASA Technical Reports Server (NTRS)

    Morgan, Gene E.; Nguyen, Francis H.

    1991-01-01

    Electrical-resistance spot-welding gun designed to produce more welds per unit time by decreasing technician's effort and fatigue. Vacuum cups on frame secure welding gun to workpiece while compressed air drives welding tip against workpiece to make spot resistance weld. When weld completed, vacuum in frame cups released so frame and gun moved to position of next spot weld.

  15. 7 CFR 27.93 - Bona fide spot markets.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 2 2011-01-01 2011-01-01 false Bona fide spot markets. 27.93 Section 27.93... REGULATIONS COTTON CLASSIFICATION UNDER COTTON FUTURES LEGISLATION Regulations Spot Markets § 27.93 Bona fide spot markets. The following markets have been determined, after investigation, and are...

  16. 7 CFR 27.93 - Bona fide spot markets.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Bona fide spot markets. 27.93 Section 27.93... REGULATIONS COTTON CLASSIFICATION UNDER COTTON FUTURES LEGISLATION Regulations Spot Markets § 27.93 Bona fide spot markets. The following markets have been determined, after investigation, and are...

  17. 21 CFR 892.1670 - Spot-film device.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Spot-film device. 892.1670 Section 892.1670 Food... DEVICES RADIOLOGY DEVICES Diagnostic Devices § 892.1670 Spot-film device. (a) Identification. A spot-film... medical purposes to position a radiographic film cassette to obtain radiographs during fluoroscopy....

  18. 21 CFR 892.1670 - Spot-film device.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Spot-film device. 892.1670 Section 892.1670 Food... DEVICES RADIOLOGY DEVICES Diagnostic Devices § 892.1670 Spot-film device. (a) Identification. A spot-film... medical purposes to position a radiographic film cassette to obtain radiographs during fluoroscopy....

  19. 21 CFR 892.1670 - Spot-film device.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Spot-film device. 892.1670 Section 892.1670 Food... DEVICES RADIOLOGY DEVICES Diagnostic Devices § 892.1670 Spot-film device. (a) Identification. A spot-film... medical purposes to position a radiographic film cassette to obtain radiographs during fluoroscopy....

  20. 21 CFR 892.1670 - Spot-film device.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Spot-film device. 892.1670 Section 892.1670 Food... DEVICES RADIOLOGY DEVICES Diagnostic Devices § 892.1670 Spot-film device. (a) Identification. A spot-film... medical purposes to position a radiographic film cassette to obtain radiographs during fluoroscopy....

  1. 21 CFR 892.1670 - Spot-film device.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Spot-film device. 892.1670 Section 892.1670 Food... DEVICES RADIOLOGY DEVICES Diagnostic Devices § 892.1670 Spot-film device. (a) Identification. A spot-film... medical purposes to position a radiographic film cassette to obtain radiographs during fluoroscopy....

  2. On the Spreading Rate and Structure of a Turbulent Spot.

    DTIC Science & Technology

    1980-03-19

    ensemble-averaged spot. The particle trajectories led CCD to conclude that there are two vortex structures associated with an ensembled averaged spot . One transverse...virtual origin for the spot . One may chose the intersec- tion of _Z and a for the virtual origin because these loci represent the ex- treme

  3. Turner Syndrome

    MedlinePlus

    Turner syndrome is a genetic disorder that affects a girl's development. The cause is a missing or incomplete ... t work properly. Other physical features typical of Turner syndrome are Short, "webbed" neck with folds of skin ...

  4. Alport syndrome

    MedlinePlus

    ... Autosomal dominant Alport syndrome (ADAS) -- This is the rarest type. Males and females have equally severe disease. Symptoms KIDNEYS With all types of Alport syndrome the kidneys are affected. The tiny blood vessels in the glomeruli of the kidneys are ...

  5. Reye syndrome

    MedlinePlus

    ... syndrome has occurred in children who were given aspirin when they had chickenpox or the flu. Reye syndrome has become very rare. This is because aspirin is no longer recommended for routine use in ...

  6. Rett Syndrome

    MedlinePlus

    Rett syndrome is a rare genetic disease that causes developmental and nervous system problems, mostly in girls. It's related to autism spectrum disorder. Babies with Rett syndrome seem to grow and develop normally at first. ...

  7. Tourette Syndrome

    MedlinePlus

    ... will order several other tests like blood tests, EEG, and brain scans. How Is Tourette Syndrome Treated? ... connected to Tourette syndrome, like ADHD and anxiety. Stress or being upset can make the tics worse, ...

  8. LEOPARD syndrome

    MedlinePlus

    LEOPARD syndrome is a very rare inherited disorder in which there are problems with the skin, face, ... LEOPARD syndrome is inherited as an autosomal dominant trait. This means the person only needs the abnormal ...

  9. Myelodysplastic Syndromes

    MedlinePlus

    ... help with blood clotting. If you have a myelodysplastic syndrome, the stem cells do not mature into healthy ... can lead to infection, anemia, or easy bleeding. Myelodysplastic syndromes often do not cause early symptoms and are ...

  10. Marfan Syndrome

    MedlinePlus

    ... Like for Kids With Marfan Syndrome? en español Síndrome de Marfan Evan couldn't wait for school ... for Marfan syndrome runs in families, getting passed down to children from parents who have the disease. ...

  11. Edwards' syndrome.

    PubMed

    Crawford, Doreen; Dearmun, Annette

    2016-12-08

    Edwards' syndrome is a serious genetic condition that affects fetal cellular functions, tissue development and organogenesis. Most infants with the syndrome are female, but there is no race predominance.

  12. Proteus Syndrome

    MedlinePlus

    ... Donate Cash Donation Life Insurance Gift Matching Gift Stock Gift Sunshine Society Contact Privacy Policy Proteus Syndrome ... approved by the Proteus Syndrome Foundation Assessment and management of the orthopedic and other complications of Proteus ...

  13. Apert Syndrome.

    PubMed

    Datta, Saikat; Saha, Sandip; Kar, Arnab; Mondal, Souvonik; Basu, Syamantak

    2014-09-01

    Apert syndrome is one of the craniosynostosis syndromes which, due to its association with other skeletal anomalies, is also known as acrocephalosyndactyly. It is a rare congenital anomaly which stands out from other craniosynostosis due to its characteristic skeletal presentations.

  14. Spot foreign exchange market and time series

    NASA Astrophysics Data System (ADS)

    Petroni, F.; Serva, M.

    2003-08-01

    We investigate high frequency price dynamics in foreign exchange market using data from Reuters information system (the dataset has been provided to us by Olsen and Associates). In our analysis we show that a naïve approach to the definition of price (for example using the spot mid price) may lead to wrong conclusions on price behavior as for example the presence of short term correlations for returns. For this purpose we introduce an algorithm which only uses the non arbitrage principle to estimate real prices from the spot ones. The new definition leads to returns which are not affected by spurious correlations. Furthermore, any apparent information (defined by using Shannon entropy) contained in the data disappears.

  15. Sweet spot supersymmetry and composite messengers

    NASA Astrophysics Data System (ADS)

    Ibe, Masahiro; Kitano, Ryuichiro

    2008-05-01

    Sweet spot supersymmetry is a phenomenological effective Lagrangian of weak scale supersymmetry with a certain set of natural assumptions. This framework is designed to avoid problems in low-energy phenomenology and cosmology of supersymmetric models. We discuss a class of dynamical models of supersymmetry breaking and its mediation, whose low-energy effective description falls into this framework. Hadron fields in the dynamical models play a role of the messengers of the supersymmetry breaking. As is always true in the models of the sweet spot supersymmetry, the messenger scale is predicted to be 105 GeV ≲Mmess ≲1010 GeV. Various values of the effective number of messenger fields Nmess are possible depending on the choice of the gauge group.

  16. Sweet Spot Supersymmetry and Composite Messengers

    SciTech Connect

    Ibe, Masahiro; Kitano, Ryuichiro

    2007-10-30

    Sweet spot supersymmetry is a phenomenologically and cosmologically perfect framework to realize a supersymmetric world at short distance. We discuss a class of dynamical models of supersymmetry breaking and its mediation whose low-energy effective description falls into this framework. Hadron fields in the dynamical models play a role of the messengers of the supersymmetry breaking. As is always true in the models of the sweet spot supersymmetry, the messenger scale is predicted to be 10{sup 5} GeV {approx}< M{sub mess} {approx}< 10{sup 10} GeV. Various values of the effective number of messenger fields N{sub mess} are possible depending on the choice of the gauge group.

  17. Dynamically variable spot size laser system

    NASA Technical Reports Server (NTRS)

    Gradl, Paul R. (Inventor); Hurst, John F. (Inventor); Middleton, James R. (Inventor)

    2012-01-01

    A Dynamically Variable Spot Size (DVSS) laser system for bonding metal components includes an elongated housing containing a light entry aperture coupled to a laser beam transmission cable and a light exit aperture. A plurality of lenses contained within the housing focus a laser beam from the light entry aperture through the light exit aperture. The lenses may be dynamically adjusted to vary the spot size of the laser. A plurality of interoperable safety devices, including a manually depressible interlock switch, an internal proximity sensor, a remotely operated potentiometer, a remotely activated toggle and a power supply interlock, prevent activation of the laser and DVSS laser system if each safety device does not provide a closed circuit. The remotely operated potentiometer also provides continuous variability in laser energy output.

  18. A case of Rocky Mountain spotted fever.

    PubMed

    Rubel, Barry S

    2007-01-01

    Rocky Mountain spotted fever is a serious, generalized infection that is spread to humans through the bite of infected ticks. It can be lethal but it is curable. The disease gets its name from the Rocky Mountain region where it was first identified in 1896. The fever is caused by the bacterium Rickettsia rickettsii and is maintained in nature in a complex life cycle involving ticks and mammals. Humans are considered to be accidental hosts and are not involved in the natural transmission cycle of this pathogen. The author examined a 47-year-old woman during a periodic recall appointment. The patient had no dental problems other than the need for routine prophylaxis but mentioned a recent problem with swelling of her extremities with an accompanying rash and general malaise and soreness in her neck region. Tests were conducted and a diagnosis of Rocky Mountain spotted fever was made.

  19. Rocky Mountain spotted fever: a clinician's dilemma.

    PubMed

    Masters, Edwin J; Olson, Gary S; Weiner, Scott J; Paddock, Christopher D

    2003-04-14

    Rocky Mountain spotted fever is still the most lethal tick-vectored illness in the United States. We examine the dilemmas facing the clinician who is evaluating the patient with possible Rocky Mountain spotted fever, with particular attention to the following 8 pitfalls in diagnosis and treatment: (1) waiting for a petechial rash to develop before diagnosis; (2) misdiagnosing as gastroenteritis; (3) discounting a diagnosis when there is no history of a tick bite; (4) using an inappropriate geographic exclusion; (5) using an inappropriate seasonal exclusion; (6) failing to treat on clinical suspicion; (7) failing to elicit an appropriate history; and (8) failing to treat with doxycycline. Early diagnosis and proper treatment save lives.

  20. Automated Spot Weld Inspection using Infrared Thermography

    SciTech Connect

    Chen, Jian; Zhang, Wei; Yu, Zhenzhen; Feng, Zhili

    2012-01-01

    An automated non-contact and non-destructive resistance spot weld inspection system based on infrared (IR) thermography was developed for post-weld applications. During inspection, a weld coupon was heated up by an auxiliary induction heating device from one side of the weld, while the resulting thermal waves on the other side were observed by an IR camera. The IR images were analyzed to extract a thermal signature based on normalized heating time, which was then quantitatively correlated to the spot weld nugget size. The use of normalized instead of absolute IR intensity was found to be useful in minimizing the sensitivity to the unknown surface conditions and environment interference. Application of the IR-based inspection system to different advanced high strength steels, thickness gauges and coatings were discussed.

  1. [Macrolides in the treatment of children with Mediterranean spotted fever].

    PubMed

    Cascio, Antonio; Colomba, Claudia

    2002-09-01

    Till now there is not a gold standard therapy for Mediterranean spotted fever (MSF) in children. Standard treatment for MSF is the administration of tetracycline or chloramphenicol, however both these drugs can cause significant adverse effects in children (tetracyclines can cause staining of teeth, chloramphenicol severe hematological adverse events such as aplastic anemia, gray baby syndrome and hemolytic anemia in patients with the Mediterranean form of G6PD deficiency). We conducted two randomized clinical trials; the first compared clarithromycin versus chloramphenicol: mean time to defervescence was 36.7 +/- 18.1 h in the clarithromycin group and 47.1+/- 21.9 h in the chloramphenicol group (P= 0.047). The second trial compared clarithromycin versus azithromycin and did not show any statistically significant difference: mean time to defervescence was 46.2 +/- 36.4 h in the clarithromycin group and 39.3 +/- 31.3 h in the azithromycin group (P= 0.34). On the basis of these studies we think that clarithromycin and azithromycin could constitute an acceptable alternative to chloramphenicol and to tetracyclines for the treatment of MSF in children

  2. Strong transverse fields in delta-spots

    NASA Technical Reports Server (NTRS)

    Zirin, Harold; Wang, Haimin

    1993-01-01

    Spectroscopic measurements of the strength and direction of transverse magnetic fields in six delta-spots are presented. The field direction is determined by the relative strength of the pi- and sigma-components at different polarizer orientations, and is, with one exception, parallel to the neutral line and as strong as the umbral field. Field strengths determined by line splitting are as high as 3980 G.

  3. Does alprostadil cream hit the spot?

    PubMed

    2015-02-01

    Alprostadil, a prostaglandin, has been marketed for many years as a urethral stick and an intracavernous injection for the treatment of erectile dysfunction.(1) It is now available in the form of a cream (Vitaros-Takeda). Adverts for the product declare: "Sex with no pills, pellets or needles. Spot on." In this article, we consider the evidence for alprostadil cream, and its place in the management of erectile dysfunction.

  4. First- and second-order Poisson spots

    NASA Astrophysics Data System (ADS)

    Kelly, William R.; Shirley, Eric L.; Migdall, Alan L.; Polyakov, Sergey V.; Hendrix, Kurt

    2009-08-01

    Although Thomas Young is generally given credit for being the first to provide evidence against Newton's corpuscular theory of light, it was Augustin Fresnel who first stated the modern theory of diffraction. We review the history surrounding Fresnel's 1818 paper and the role of the Poisson spot in the associated controversy. We next discuss the boundary-diffraction-wave approach to calculating diffraction effects and show how it can reduce the complexity of calculating diffraction patterns. We briefly discuss a generalization of this approach that reduces the dimensionality of integrals needed to calculate the complete diffraction pattern of any order diffraction effect. We repeat earlier demonstrations of the conventional Poisson spot and discuss an experimental setup for demonstrating an analogous phenomenon that we call a "second-order Poisson spot." Several features of the diffraction pattern can be explained simply by considering the path lengths of singly and doubly bent paths and distinguishing between first- and second-order diffraction effects related to such paths, respectively.

  5. Hot spot conditions during cavitation in water

    SciTech Connect

    Didenko, Y.T.; McNamara, W.B. III; Suslick, K.S.

    1999-06-23

    Liquids irradiated with high-intensity ultrasound undergo acoustic cavitation--the formation, growth, and implosive collapse of bubbles. The energy stored during the growth of the bubble in the rarefaction phase of the acoustic field is released when the bubble violently collapses in the compression phase of the acoustic field, as acoustic noise, shock waves, chemical reactions, and the emission of light (sonoluminescence, SL). This violent collapse is predicted to generate a hot spot of thousands of Kelvin within the bubble, but, to date, there have been only a limited number of experimental measurements of the temperature of this hot spot. Although the SL of water has been studied for more than 50 years, the effective hot spot temperature during aqueous cavitation remains unresolved. Given the importance of aqueous cavitation (sonography and bioeffects of ultrasound, sonochemical remediation of aqueous pollutants, synthetic applications of sonochemistry, etc.), the authors decided to apply previous spectroscopic analysis of SL of nonaqueous liquids to aqueous solutions doped with small amounts of hydrocarbons. The authors have collected and analyzed excited-state C{sub 2} NBSL (multi-bubble sonoluminescence, light emission from the clouds of cavitating bubbles) spectra from mixtures of organic liquids in water at 20 kHz and find an effective emission temperature of 4,300 {+-} 200 K.

  6. Velocardiofacial Syndrome

    ERIC Educational Resources Information Center

    Gothelf, Doron; Frisch, Amos; Michaelovsky, Elena; Weizman, Abraham; Shprintzen, Robert J.

    2009-01-01

    Velocardiofacial syndrome (VCFS), also known as DiGeorge, conotruncal anomaly face, and Cayler syndromes, is caused by a microdeletion in the long arm of Chromosome 22. We review the history of the syndrome from the first clinical reports almost half a century ago to the current intriguing molecular findings associating genes from the…

  7. Fraser syndrome.

    PubMed

    Kalpana Kumari, M K; Kamath, Sulata; Mysorekar, Vijaya V; Nandini, G

    2008-01-01

    Fraser syndrome or cryptophthalmos is a rare autosomal recessive disorder characterized by major features such as cryptophthalmos, syndactyly and abnormal genitalia. The diagnosis of this syndrome can be made on clinical examination and perinatal autopsy. We present the autopsy findings of a rare case of Fraser syndrome in a male infant.

  8. Friction Stir Spot Welding of Advanced High Strength Steels

    SciTech Connect

    Hovanski, Yuri; Grant, Glenn J.; Santella, M. L.

    2009-11-13

    Friction stir spot welding techniques were developed to successfully join several advanced high strength steels. Two distinct tool materials were evaluated to determine the effect of tool materials on the process parameters and joint properties. Welds were characterized primarily via lap shear, microhardness, and optical microscopy. Friction stir spot welds were compared to the resistance spot welds in similar strength alloys by using the AWS standard for resistance spot welding high strength steels. As further comparison, a primitive cost comparison between the two joining processes was developed, which included an evaluation of the future cost prospects of friction stir spot welding in advanced high strength steels.

  9. Characterizing hot spots throughout the catchment

    NASA Astrophysics Data System (ADS)

    Welti, N.; Lockington, D.; Jakeman, T.; Hunt, R.

    2012-04-01

    Few catchments in the world are left truly undisturbed. Rather, they are under anthropogenic stress for a variety of reasons ranging from climate forcing to meeting the basic water allocation needs of the population. Reduction in the number of inundation areas has significantly decreased the nutrient and organic matter retention capacity along the river corridor, with major consequences for the both the riverine and coastal ecosystems. Cumulative stress may build up to a "tipping point" which can cause a change or set of changes which could occur non-linearly. In order to mitigate the environmental stress on these ecosystems, management plans are created to balance the needs of the dependent populations and those of ecology. While these catchment-wide plans aim to improve the ecological function of aquatic areas over the large scale, this sledge-hammer approach ignores the inherent heterogeneity in the catchment. Societal (and policy) decisions involve more than abiotic quantification of water storage and flow. A more encompassing ecohydrological view facilitates a more rounded policy framework that has flexibility to accommodate multiple social drivers, and one that can accommodate an "ecosystem improvement" rather than single species improvement. Not every spot in the landscape is equally valuable for specific societal values. Areas of high activity may provide the resilience capacity necessary to prevent catastrophic changes. In times of ecological instability, ecosystem resilience is of paramount importance in maintaining essential ecosystem services. Hot spots of biogeochemical cycling will occur where unique situations arise, such as areas of surface and groundwater interaction, creating spots of localized, high activity. In order to understand the systems' potential to support various habitat niches in the large scale, the identification of specific hot spots or hot moments is necessary. A basal understanding of the concurrent biogeochemical cycles enables

  10. Surgical syndromes of the hypothalamus.

    PubMed

    Carmel, P W

    1980-01-01

    The clinical syndromes described with lesions of the hypothalamus are summarized in Table 9.5-9.7. The anterior hypothalamic syndrome consists of insomnia and loss of thirst regulatory mechanisms. In occasional larger lesions which interrupt the output from the supraoptic and paraventricular nuclei, diabetes insipidus has been noticed. In the tuberal region of the hypothalamus the most prominent findings are those that are caused by the disruption of the final common pathway to the pituitary. This results in endocrinopathy, most often the syndrome originally reported by Frohlich, with failure of sexual maturation and obesity. In the tuberal region, differences between lesions of the medial and lateral portions are quite marked. Medial lesions result in obesity while bilateral lesions result in anorexia and emaciation. The diencephalic syndrome of infancy with it's severe emaciation in young years and obesity in later years clearly indicates a different organizational pattern in the neonatal hypothalamus. Emotional disorders may be seen with lesions either in the medial or lateral hypothalamus at the tuberal level. Finally, in the posterior hypothalamic region, which includes the greatest effector apparatus, hypersomnia, apathy, and poikilothermia have been reported. Emotional disturbances and the Wernicke-Korsakoff syndrome also seemed to be associated with lesions in this area. The hypothalamus remains the single most important integrator of vegetative and endocrinologic regulation of the body. Cushing said of the hypothalamus, "here in this hidden spot, almost to be covered with a thumb nail, lies the very main spring of primitive existence: vegetative, emotional and reproductive".

  11. Star spot location estimation using Kalman filter for star tracker.

    PubMed

    Liu, Hai-bo; Yang, Jian-kun; Wang, Jiong-qi; Tan, Ji-chun; Li, Xiu-jian

    2011-04-20

    Star pattern recognition and attitude determination accuracy is highly dependent on star spot location accuracy for the star tracker. A star spot location estimation approach with the Kalman filter for a star tracker has been proposed, which consists of three steps. In the proposed approach, the approximate locations of the star spots in successive frames are predicted first; then the measurement star spot locations are achieved by defining a series of small windows around each predictive star spot location. Finally, the star spot locations are updated by the designed Kalman filter. To confirm the proposed star spot location estimation approach, the simulations based on the orbit data of the CHAMP satellite and the real guide star catalog are performed. The simulation results indicate that the proposed approach can filter out noises from the measurements remarkably if the sampling frequency is sufficient.

  12. Stability of the Boundary Layer and the Spot

    NASA Technical Reports Server (NTRS)

    Wygnanski, I.

    2007-01-01

    The similarity among turbulent spots observed in various transition experiments, and the rate in which they contaminate the surrounding laminar boundary layer is only cursory. The shape of the spot depends on the Reynolds number of the surrounding boundary layer and on the pressure gradient to which it and the surrounding laminar flow are exposed. The propagation speeds of the spot boundaries depend, in addition, on the location from which the spot originated and do not simply scale with the local free stream velocity. The understanding of the manner in which the turbulent manner in which the turbulent spot destabilizes the surrounding, vortical fluid is a key to the understanding of the transition process. We therefore turned to detailed observations near the spot boundaries in general and near the spanwise tip of the spot in particular.

  13. [Autoinflammatory syndrome].

    PubMed

    Ida, Hiroaki; Eguchi, Katsumi

    2009-03-01

    The autoinflammatory syndromes include a group of inherited diseases that are characterized by 1) seemingly unprovoked episodes of systemic inflammations, 2) absence of high titer of autoantibody or auto-reactive T cell, and 3) inborn error of innate immunity. In this article, we will focus on the clinical features, the pathogenesis related the genetic defects, and the therapeutic strategies in the representative disorders including familial Mediterranean fever (FMF), TNF receptor associated periodic syndrome (TRAPS), cryopyrin-associated periodic syndrome (CAPS), hyper-IgD with periodic fever syndrome (HIDS), syndrome of pyogenic arthritis with pyoderma gangrenosum and acne (PAPA), and Blau syndrome. Recent advances in genetics and molecular biology have proceeded our understanding of the pathogenesis of autoinflammatory syndromes.

  14. Hurler syndrome with a tuft of hair.

    PubMed

    Behera, Binodini; Jena, D K; Chhetia, R; Vijayashree, J

    2006-01-01

    A 2-year-old girl presented with coarse, thick hairy skin all over the body, a tuft of hair in the parietal region, coarse facial features and a prominent forehead with a large tongue, hepatosplenomegaly and skeletal deformities. Mucopolysaccharides excretion spot test of the urine was positive; and an assay for glycosaminoglycans in the urine was also high, which confirmed the clinical diagnosis of Hurler syndrome. We present this rare case to discuss the possibility of the association of mental retardation with a tuft of hair in this syndrome.

  15. Detection of a non-Gaussian spot in WMAP

    NASA Astrophysics Data System (ADS)

    Cruz, M.; Martínez-González, E.; Vielva, P.; Cayón, L.

    2005-01-01

    An extremely cold and big spot in the Wilkinson Microwave Anisotropy Probe (WMAP) 1-yr data is analysed. Our work is a continuation of a previous paper by Vielva et al. that reported the detection of non-Gaussianity, with a method based on the spherical Mexican hat wavelet (SMHW) technique. We study the spots at different thresholds on the SMHW coefficient maps, considering six estimators, namely the number of maxima, the number of minima, the numbers of hot and cold spots, and the number of pixels of those spots. At SMHW scales around 4° (10° on the sky), the data deviate from Gaussianity. The analysis is performed on all of the sky, the Northern and Southern hemispheres, and on four regions covering all of the sky. A cold spot at (b=-57°,l= 209°) is found to be the source of this non-Gaussian signature. We compare the spots of our data with 10000 Gaussian simulations, and conclude that only around 0.2 per cent of them present such a cold spot. Excluding this spot, the remaining map is compatible with Gaussianity, and even the excess of kurtosis in the paper by Vielva et al. is found to be due exclusively to this spot. Finally, we study whether the spot causing the observed deviation from Gaussianity could be generated by systematics or foregrounds. None of them seem to be responsible for the non-Gaussian detection.

  16. The fractal nature of vacuum arc cathode spots

    SciTech Connect

    Anders, Andre

    2005-05-27

    Cathode spot phenomena show many features of fractals, for example self-similar patterns in the emitted light and arc erosion traces. Although there have been hints on the fractal nature of cathode spots in the literature, the fractal approach to spot interpretation is underutilized. In this work, a brief review of spot properties is given, touching the differences between spot type 1 (on cathodes surfaces with dielectric layers) and spot type 2 (on metallic, clean surfaces) as well as the known spot fragment or cell structure. The basic properties of self-similarity, power laws, random colored noise, and fractals are introduced. Several points of evidence for the fractal nature of spots are provided. Specifically power laws are identified as signature of fractal properties, such as spectral power of noisy arc parameters (ion current, arc voltage, etc) obtained by fast Fourier transform. It is shown that fractal properties can be observed down to the cutoff by measurement resolution or occurrence of elementary steps in physical processes. Random walk models of cathode spot motion are well established: they go asymptotically to Brownian motion for infinitesimal step width. The power spectrum of the arc voltage noise falls as 1/f {sup 2}, where f is frequency, supporting a fractal spot model associated with Brownian motion.

  17. Introgression and dispersal among spotted owl (Strix occidentalis) subspecies

    PubMed Central

    Funk, W Chris; Forsman, Eric D; Mullins, Thomas D; Haig, Susan M

    2008-01-01

    Abstract Population genetics plays an increasingly important role in the conservation and management of declining species, particularly for defining taxonomic units. Subspecies are recognized by several conservation organizations and countries and receive legal protection under the US Endangered Species Act (ESA). Two subspecies of spotted owls, northern (Strix occidentalis caurina) and Mexican (S. o. lucida) spotted owls, are ESA-listed as threatened, but the California (S. o. occidentalis) spotted owl is not listed. Thus, determining the boundaries of these subspecies is critical for effective enforcement of the ESA. We tested the validity of previously recognized spotted owl subspecies by analysing 394 spotted owls at 10 microsatellite loci. We also tested whether northern and California spotted owls hybridize as suggested by previous mitochondrial DNA studies. Our results supported current recognition of three subspecies. We also found bi-directional hybridization and dispersal between northern and California spotted owls centered in southern Oregon and northern California. Surprisingly, we also detected introgression of Mexican spotted owls into the range of northern spotted owls, primarily in the northern part of the subspecies’ range in Washington, indicating long-distance dispersal of Mexican spotted owls. We conclude with a discussion of the conservation implications of our study. PMID:25567499

  18. False Color Mosaic Great Red Spot

    NASA Technical Reports Server (NTRS)

    1996-01-01

    False color representation of Jupiter's Great Red Spot (GRS) taken through three different near-infrared filters of the Galileo imaging system and processed to reveal cloud top height. Images taken through Galileo's near-infrared filters record sunlight beyond the visible range that penetrates to different depths in Jupiter's atmosphere before being reflected by clouds. The Great Red Spot appears pink and the surrounding region blue because of the particular color coding used in this representation. Light reflected by Jupiter at a wavelength (886 nm) where methane strongly absorbs is shown in red. Due to this absorption, only high clouds can reflect sunlight in this wavelength. Reflected light at a wavelength (732 nm) where methane absorbs less strongly is shown in green. Lower clouds can reflect sunlight in this wavelength. Reflected light at a wavelength (757 nm) where there are essentially no absorbers in the Jovian atmosphere is shown in blue: This light is reflected from the deepest clouds. Thus, the color of a cloud in this image indicates its height. Blue or black areas are deep clouds; pink areas are high, thin hazes; white areas are high, thick clouds. This image shows the Great Red Spot to be relatively high, as are some smaller clouds to the northeast and northwest that are surprisingly like towering thunderstorms found on Earth. The deepest clouds are in the collar surrounding the Great Red Spot, and also just to the northwest of the high (bright) cloud in the northwest corner of the image. Preliminary modeling shows these cloud heights vary over 30 km in altitude. This mosaic, of eighteen images (6 in each filter) taken over a 6 minute interval during the second GRS observing sequence on June 26, 1996, has been map-projected to a uniform grid of latitude and longitude. North is at the top.

    Launched in October 1989, Galileo entered orbit around Jupiter on December 7, 1995. The spacecraft's mission is to conduct detailed studies of the giant planet

  19. A Magic Spot in Genome Maintenance.

    PubMed

    Rasouly, Aviram; Pani, Bibhusita; Nudler, Evgeny

    2017-01-01

    Nucleotide excision repair (NER) is the key DNA repair system that eliminates the majority of DNA helix-distorting lesions. RNA polymerase (RNAP) expedites the recognition of DNA damage by NER components via transcription-coupled DNA repair (TCR). In bacteria, a modified nucleotide ppGpp ('magic spot') is a pleiotropic second messenger that mediates the response to nutrient deficiencies by altering the initiation properties of RNAP. In this review, we discuss newly elucidated roles of guanosine 5'-diphosphate 3'-diphosphate (ppGpp) in transcription elongation that couple this alarmone to DNA damage repair and maintenance.

  20. What's new in Rocky Mountain spotted fever?

    PubMed

    Chen, Luke F; Sexton, Daniel J

    2008-09-01

    Rocky Mountain spotted fever (RMSF) remains an important illness despite an effective therapy because it is difficult to diagnose and is capable of producing a fatal outcome. The pathogenesis of RMSF remains, in large part, an enigma. However, recent research has helped shed light on this mystery. Importantly, the diagnosis of RMSF must be considered in all febrile patients who have known or possible exposure to ticks, especially if they live in or have traveled to endemic regions during warmer months. Decisions about giving empiric therapy to such patients are difficult and require skill and careful judgement.

  1. Roentgenographic abnormalities in Rocky Mountain Spotted Fever.

    PubMed

    McCook, T A; Briley, C; Ravin, C E

    1982-02-01

    Rock Mountain spotted fever (RMSF) is a tick-borne rickettsial disease which produces a widespread vasculitis. A mortality of 7% to 13% has been reported in the United States which is due at least in part to delay in diagnosis and appropriate treatment. The classic features of this disease include a history of tick bite with the clinical presentation of skin rash and fever in association with thrombocytopenia. Few reports have emphasized the radiologic chest abnormalities in this disease or their relationship to thrombocytopenia. We review 70 cases of RMSF with abnormal roentgenographic features and their pathologic correlation.

  2. Studies on Typhus and Spotted Fever.

    DTIC Science & Technology

    1980-02-01

    by block ntmber) Rickettsia , R. prowazekii, R. mooseri, typhus, R. rickettsii , Rocky Mountain spotted fever, Rochalimaea guintana, trench fever, DNA...which appear to be the major target cells for the obligate intracellular parasitic bacteria of the genus Rickettsia . This would constitute a unique...Yes e Rickettsia rickettsii Sheila Smith Human GP2/E6/TC3/E2/TC4/E6 Yes f Ripley Human GP1/CE1/E1 Yes g Rickettsia tsutsugamushi Gilliam E141/M2/E2 No h

  3. Real prices from spot foreign exchange market

    NASA Astrophysics Data System (ADS)

    Petroni, Filippo; Serva, Maurizio

    2004-12-01

    In this work we discuss the problem of price definition when using high frequency foreign exchange data. If one uses the spot mid price a strong autocorrelation of returns, at one lag, is found which is only due to microstructure effect and does not capture the real behavior of price dynamics. This autocorrelation increases the intraday volatility estimated from this type of data. To solve this problem we introduce an algorithm which is able, by using the no-arbitrage principle, of eliminating every microstructure effects.

  4. [Autoinflammatory syndromes].

    PubMed

    Lamprecht, P; Gross, W L

    2009-06-01

    In its strict sense, the term "autoinflammatory syndromes" comprises the hereditary periodic fever syndromes (HPF), which are caused by mutations of pattern-recognition receptors (PRR) and perturbations of the cytokine balance. These include the crypyrinopathies, familial Mediterranean fever, TNF-receptor associated periodic fever syndrome (TRAPS), hyper-IgD and periodic syndrome (HIDS), pyogenic sterile arthritis, pyoderma gangrenosum and acne (PAPA) syndrome, NALP12-HPF, and the Blau syndrome. The diseases are characterized by spontaneous activation of cells of the innate immunity in the absence of ligands. Autoantibodies are usually not found. HPF clinically present with recurrent fever episodes and inflammation, especially of serosal and synovial interfaces and the skin. Intriguingly, PRR-mediated autoinflammtory mechanisms also play a role in a number of chronic inflammatory and autoimmune diseases.

  5. Inducibility of somatic colour and white spots in the mammalian spot test.

    PubMed

    Peter, S

    1980-03-01

    F1 embryos of the C57BL/6JHan x T-Stock were exposed to utero to 5, 10, 15 and 45 mg/kg b.w. of cyclophosphamide (CP) s.c. on the 10th day of pregnancy. 3-5 weeks post-partum the offspring were examined for the appearance of recessive coat-colour mosaics and white midventral spots (WMVS). The frequencies of the coloured spots were 0.93 and 2.59% at 5- and 10-mg/kg doses of CP, resp. These frequencies were dose-dependent, but the difference between the doses was not significant. No coloured spots were obtained with CP at 15 and 45 mg/kg. The frequencies of WMVS were much higher than those of coloured spots (1.40, 6.03 and 51.16%, resp.). The differences were highly significant. The reduction in the number of offspring compared with the raio of the offspring/female of the control (control = 100%) were 16.16, 22.72, 72.48 and 100% at the different doses of CP (5, 10, 15 and 45 mg/kg b.w.).

  6. Ammonium Hydrosulfide and Jupiter's Great Red Spot

    NASA Astrophysics Data System (ADS)

    Loeffler, M. J.; Hudson, R.; Chanover, N.; Simon, A. A.

    2014-12-01

    The color and composition of Jupiter's Great Red Spot (GRS) has been debated for more than a century. While there are numerous hypotheses for the origin of Jupiter's GRS, recent work suggests that the GRS's color could originate from multiple components (Carlson et al., 2012; Simon et al., submitted). In light of this, we have recently begun conducting in situ laboratory experiments that test whether ammonium hydrosulfide, NH4SH, or its radiation decomposition products contribute to the GRS spectrum. In this presentation, we will discuss some of our most recent results, where we have studied the stability of NH4SH samples as a function of temperature using infrared and mass spectrometry. Funding for this work has been provided by NASA's Planetary Atmospheres and Outer Planets Research programs. ReferencesCarlson, R. W., K. H. Baines, M. S. Anderson, G. Filacchione. Chromophores from photolyzed ammonia reacting with acetylene: Application to Jupiter's Great Red Spot, DPS, 44, 2012. Simon, A. A., J. Legarreta, F. Sanz-Requena, S. Perez-Hoyos, E. Garcia-Melendo, R. W. Carlson. Spectral Comparison and Stability of Red Regions on Jupiter. J. Geophys. Res. - Planets, submitted.

  7. SPOT4 Operational Control Center (CMP)

    NASA Technical Reports Server (NTRS)

    Zaouche, G.

    1993-01-01

    CNES(F) is responsible for the development of a new generation of Operational Control Center (CMP) which will operate the new heliosynchronous remote sensing satellite (SPOT4). This Operational Control Center takes large benefit from the experience of the first generation of control center and from the recent advances in computer technology and standards. The CMP is designed for operating two satellites all the same time with a reduced pool of controllers. The architecture of this CMP is simple, robust, and flexible, since it is based on powerful distributed workstations interconnected through an Ethernet LAN. The application software uses modern and formal software engineering methods, in order to improve quality and reliability, and facilitate maintenance. This software is table driven so it can be easily adapted to other operational needs. Operation tasks are automated to the maximum extent, so that it could be possible to operate the CMP automatically with very limited human interference for supervision and decision making. This paper provides an overview of the SPOTS mission and associated ground segment. It also details the CMP, its functions, and its software and hardware architecture.

  8. Beyond Binarity: Spots, Pulsations, and Triple Systems

    NASA Astrophysics Data System (ADS)

    Johnston, Cole; Prsa, A.

    2014-01-01

    We use the Kepler Eclipsing Binary Catalog (Prsa et al. 2011) to find and explore previously unstudied intrinsic stellar variability and stellar multiplicity. All but the highest-amplitude intrinsic variation in these systems is dominated by the eclipsing binary signature, however by fitting a physical model to the eclipsing binary signal and then subtracting this model from the lightcurve, we effectively remove binary effects and can search the residuals for other sources of variability. Using 120 stars for our sample, observed at a 1-min cadence by NASA's Kepler satellite (Borucki et al. 2009), we find low amplitude spot variation, pulsations, and background eclipsing binary stars. Frequencies derived from the spot variations and pulsations provide us with information on the rotation rates, internal structure and physical parameters of the stars that comprise each system. Using frequency and period relations derived by Tassoul (1980), we identify g-mode and p-mode pulsations from the derived signals. We apply asteroseismic methods to interpret these signals and determine the radii and masses of the system components, differential surface and interior rotation, and evolutionary state of these stars. Binary star modeling of these systems yields independent values of the masses, radii, and temperatures of both components, as well as any tidal deformation that may occur. By coupling these approaches, we aim able to construct a fully consistent model of the systems that undergo these variations. We highlight systems of particular interest and discuss frequently observed features in the power spectra.

  9. Laser induced single spot oxidation of titanium

    NASA Astrophysics Data System (ADS)

    Jwad, Tahseen; Deng, Sunan; Butt, Haider; Dimov, S.

    2016-11-01

    Titanium oxides have a wide range of applications in industry, and they can be formed on pure titanium using different methods. Laser-induced oxidation is one of the most reliable methods due to its controllability and selectivity. Colour marking is one of the main applications of the oxidation process. However, the colourizing process based on laser scanning strategies is limited by the relative large processing area in comparison to the beam size. Single spot oxidation of titanium substrates is proposed in this research in order to increase the resolution of the processed area and also to address the requirements of potential new applications. The method is applied to produce oxide films with different thicknesses and hence colours on titanium substrates. High resolution colour image is imprinted on a sheet of pure titanium by converting its pixels' colours into laser parameter settings. Optical and morphological periodic surface structures are also produced by an array of oxide spots and then analysed. Two colours have been coded into one field and the dependencies of the reflected colours on incident and azimuthal angles of the light are discussed. The findings are of interest to a range of application areas, as they can be used to imprint optical devices such as diffusers and Fresnel lenses on metallic surfaces as well as for colour marking.

  10. Spinning-Spot Shadowless TIRF Microscopy.

    PubMed

    Ellefsen, Kyle L; Dynes, Joseph L; Parker, Ian

    2015-01-01

    Total internal reflection fluorescence (TIRF) microscopy is a powerful tool for visualizing near-membrane cellular structures and processes, including imaging of local Ca2+ transients with single-channel resolution. TIRF is most commonly implemented in epi-fluorescence mode, whereby laser excitation light is introduced at a spot near the periphery of the back focal plane of a high numerical aperture objective lens. However, this approach results in an irregular illumination field, owing to interference fringes and scattering and shadowing by cellular structures. We describe a simple system to circumvent these limitations, utilizing a pair of galvanometer-driven mirrors to rapidly spin the laser spot in a circle at the back focal plane of the objective lens, so that irregularities average out during each camera exposure to produce an effectively uniform field. Computer control of the mirrors enables precise scanning at 200 Hz (5ms camera exposure times) or faster, and the scan radius can be altered on a frame-by-frame basis to achieve near-simultaneous imaging in TIRF, widefield and 'skimming plane' imaging modes. We demonstrate the utility of the system for dynamic recording of local inositol trisphosphate-mediated Ca2+ signals and for imaging the redistribution of STIM and Orai proteins during store-operated Ca2+ entry. We further anticipate that it will be readily applicable for numerous other near-membrane studies, especially those involving fast dynamic processes.

  11. Spinning-Spot Shadowless TIRF Microscopy

    PubMed Central

    Ellefsen, Kyle L.; Dynes, Joseph L.; Parker, Ian

    2015-01-01

    Total internal reflection fluorescence (TIRF) microscopy is a powerful tool for visualizing near-membrane cellular structures and processes, including imaging of local Ca2+ transients with single-channel resolution. TIRF is most commonly implemented in epi-fluorescence mode, whereby laser excitation light is introduced at a spot near the periphery of the back focal plane of a high numerical aperture objective lens. However, this approach results in an irregular illumination field, owing to interference fringes and scattering and shadowing by cellular structures. We describe a simple system to circumvent these limitations, utilizing a pair of galvanometer-driven mirrors to rapidly spin the laser spot in a circle at the back focal plane of the objective lens, so that irregularities average out during each camera exposure to produce an effectively uniform field. Computer control of the mirrors enables precise scanning at 200 Hz (5ms camera exposure times) or faster, and the scan radius can be altered on a frame-by-frame basis to achieve near-simultaneous imaging in TIRF, widefield and ‘skimming plane’ imaging modes. We demonstrate the utility of the system for dynamic recording of local inositol trisphosphate-mediated Ca2+ signals and for imaging the redistribution of STIM and Orai proteins during store-operated Ca2+ entry. We further anticipate that it will be readily applicable for numerous other near-membrane studies, especially those involving fast dynamic processes. PMID:26308212

  12. Gorlin syndrome.

    PubMed

    Devi, Basanti; Behera, Binodini; Patro, Sibasish; Pattnaik, Subhransu S; Puhan, Manas R

    2013-05-01

    Gorlin Syndrome, a rare genodermatosis, otherwise known as Nevoid basal cell carcinoma syndrome (NBCCS) is a multisystem disease affecting skin, nervous system, eyes, endocrine glands, and bones. It is characterized by multiple basal cell carcinomas, palmoplantar pits, jaw cysts, and bony deformities like kyphoscoliosis and frontal bossing. We would like to report a case of Gorlin syndrome with classical features, as this is a rare genodermatosis.

  13. Overgrowth Syndromes

    PubMed Central

    Edmondson, Andrew C.; Kalish, Jennifer M.

    2015-01-01

    Numerous multiple malformation syndromes associated with pathologic overgrowth have been described and, for many, their molecular bases elucidated. This review describes the characteristic features of these overgrowth syndromes, as well as the current understanding of their molecular bases, intellectual outcomes, and cancer predispositions. We review syndromes such as Sotos, Malan, Marshall–Smith, Weaver, Simpson–Golabi–Behmel, Perlman, Bannayan–Riley–Ruvalcaba, PI3K-related, Proteus, Beckwith–Wiedemann, fibrous dysplasia, Klippel–Trenaunay–Weber, and Maffucci. PMID:27617124

  14. Spot: A Programming Language for Verified Flight Software

    NASA Technical Reports Server (NTRS)

    Bocchino, Robert L., Jr.; Gamble, Edward; Gostelow, Kim P.; Some, Raphael R.

    2014-01-01

    The C programming language is widely used for programming space flight software and other safety-critical real time systems. C, however, is far from ideal for this purpose: as is well known, it is both low-level and unsafe. This paper describes Spot, a language derived from C for programming space flight systems. Spot aims to maintain compatibility with existing C code while improving the language and supporting verification with the SPIN model checker. The major features of Spot include actor-based concurrency, distributed state with message passing and transactional updates, and annotations for testing and verification. Spot also supports domain-specific annotations for managing spacecraft state, e.g., communicating telemetry information to the ground. We describe the motivation and design rationale for Spot, give an overview of the design, provide examples of Spot's capabilities, and discuss the current status of the implementation.

  15. Not so hot "hot spots" in the oceanic mantle.

    PubMed

    Bonath, E

    1990-10-05

    Excess volcanism and crustal swelling associated with hot spots are generally attributed to thermal plumes upwelling from the mantle. This concept has been tested in the portion of the Mid-Atlantic Ridge between 34 degrees and 45 degrees (Azores hot spot). Peridotite and basalt data indicate that the upper mantle in the hot spot has undergone a high degree of melting relative to the mantle elsewhere in the North Atlantic. However, application of various geothermometers suggests that the temperature of equilibration of peridotites in the mantle was lower, or at least not higher, in the hot spot than elsewhere. The presence of H(2)O-rich metasomatized mantle domains, inferred from peridotite and basalt data, would lower the melting temperature of the hot spot mantle and thereby reconcile its high degree ofmelting with the lack of a mantle temperature anomaly. Thus, some so-called hot spots might be melting anomalies unrelated to abnormally high mantle temperature or thermal plumes.

  16. Proteus Syndrome Foundation

    MedlinePlus

    ... Gift Stock Gift Sunshine Society Contact Privacy Policy Proteus Syndrome Foundation The Proteus Syndrome Foundation , a 501c3 ... 1 Trial with ARQ 092 in Proteus Syndrome Proteus Syndrome Patient Registry The Proteus Syndrome Foundation Contact ...

  17. Bier’s spots with onset in childhood*

    PubMed Central

    Portocarrero, Larissa Karine Leite; Saraiva, Maria Isabel Ramos; Barbosa, Marcella Amaral Horta; Veronez, Isis Suga; Swiczar, Bethania Cabral Cavalli; Valente, Neusa Yuriko Sakai

    2016-01-01

    Bier spots are small, irregular, hypopigmented macules that are usually found on the arms and legs. The macules disappear when the limb is raised. Bier spots have been reported in association with a number of conditions but there is no consistent association to specific desease. Although they usually affect young adults, we report a case of Bier spots that began in childhood. As an asymptomatic and possibly transitional condition, the disease does not require treatment. PMID:28300906

  18. Nondestructive spot test method for magnesium and magnesium alloys

    NASA Technical Reports Server (NTRS)

    Wilson, M. L. (Inventor)

    1973-01-01

    A method for spot test identification of magnesium and various magnesium alloys commonly used in aerospace applications is described. The spot test identification involves color codes obtained when several drops of 3 M hydrochloric acid are placed on the surface to be tested. After approximately thirty seconds, two drops of this reacted acid is transferred to each of two depressions in a spot plate for additions of other chemicals with subsequent color changes indicating magnesium or its alloy.

  19. Dominant inheritance of overo spotting in paint horses.

    PubMed

    Bowling, A T

    1994-01-01

    Analysis of selected studbook records of the American Paint Horse Association, consisting of 687 foals sired by 13 overo stallions from non-overo mares, supports the inheritance of overo spotting as an autosomal dominant gene. More than one gene may control patterns registered as overo. Additional studies are necessary to explain the sporadic occurrence of overo spotting from nonspotted quarter horse parents and to confirm the inheritance of overo spotting in other breeds.

  20. Aerosol particle microphotography and glare-spot absorption spectroscopy.

    PubMed

    Arnold, S; Holler, S; Li, J H; Serpengüzel, A; Auffermann, W F; Hill, S C

    1995-04-01

    The relative intensities of glare spots in the image of an electrodynamically trapped aerosol droplet are measured experimentally with an aerosol particle microscope and calculated theoretically. The theoretical calculations are in good agreement with these experiments and indicate that the intensities of these spots are extremely sensitive to the imaginary part of the refractive index. Experimentally, we obtain the molecular absorption spectrum of an impurity within a droplet by recording the spectrum of an individual glare spot produced by broadband illumination.

  1. Dynamic Characterization of the CT Angiographic ‘Spot Sign’

    PubMed Central

    Chakraborty, Santanu; Alhazzaa, Mohammed; Wasserman, Jason K.; Sun, Yang Yang; Stotts, Grant; Hogan, Mathew J.; Demchuk, Andrew; Aviv, Richard I.; Dowlatshahi, Dar

    2014-01-01

    Background and purpose Standard (static) CT angiography is used to identify the intracerebral hemorrhage (ICH) spot sign. We used dynamic CT-angiography to describe spot sign characteristics and measurement parameters over 60-seconds of image acquisition. Methods We prospectively identified consecutive patients presenting with acute ICH within 4.5 hours of symptom onset, and collected whole brain dynamic CT-angiography (dCTA). Spot parameters (earliest appearance, duration, maximum Hounsfield unit (HU), time to maximum HU, time to spot diagnostic definition, spot volume and hematoma volumes) were measured using volumetric analysis software. Result We enrolled 34 patients: three were excluded due to secondary causes of ICH. Of the remaining 31 patients there were 18 females (58%) with median age 70 (range 47–86) and baseline hematoma volume 33 ml (range 0.7–103 ml). Positive dCTA spot sign was present in 13 patients (42%) visualized as an expanding 3-dimensional structure temporally evolving its morphology over the scan period. Median time to spot appearance was 21 s (range 15–35 seconds). This method allowed tracking of spots evolution until the end of venous phase (active extravasation) with median duration of 39 s (range 25–45 seconds). The average density and time to maximum density was 204HU and 30.8 s (range 23–31 s) respectively. Median time to spot diagnosis was 20.8 s using either 100 or 120HU definitions. Conclusion Dynamic CTA allows a 3-dimensional assessment of spot sign formation during acute ICH, and captured higher spot sign prevalence than previously reported. This is the first study to describe and quantify spot sign characteristics using dCTA; these can be used in ongoing and upcoming ICH studies. PMID:24594897

  2. Eye-spots in Lepidoptera attract attention in humans

    PubMed Central

    Yorzinski, Jessica L.; Platt, Michael L.; Adams, Geoffrey K.

    2015-01-01

    Many prey species exhibit defensive traits to decrease their chances of predation. Conspicuous eye-spots, concentric rings of contrasting colours, are one type of defensive trait that some species exhibit to deter predators. We examined the function of eye-spots in Lepidoptera to determine whether they are effective at deterring predators because they resemble eyes (‘eye mimicry hypothesis’) or are highly salient (‘conspicuous signal hypothesis’). We recorded the gaze behaviour of men and women as they viewed natural images of butterflies and moths as well as images in which the eye-spots of these insects were modified. The eye-spots were modified by removing them, scrambling their colours, or replacing them with elliptical or triangular shapes that had either dark or light centres. Participants were generally more likely to look at, spend more time looking at and be faster to first fixate the eye-spots of butterflies and moths that were natural compared with ones that were modified, including the elliptical eye-spots with dark centres that most resembled eyes as well as the scrambled eye-spots that had the same contrast as the natural eye-spots. Participants were most likely to look at eye-spots that were numerous, had a large surface area and were located close to the insects' heads. Participants' pupils were larger when viewing eye-spots compared with the rest of the insects' body, suggesting a greater arousal when viewing eye-spots. Our results provide some support for the conspicuous signal hypothesis (and minimal support for the eye mimicry hypothesis) and suggest that eye-spots may be effective at deterring predators because they are highly conspicuous signals that draw attention. PMID:26543589

  3. Simultaneous velocity-surface heat transfer behavior of turbulent spots

    NASA Astrophysics Data System (ADS)

    Sabatino, D.; Smith, C. R.

    2002-06-01

    The properties of artificially initiated turbulent spots in a heated laminar boundary layer were investigated in a water channel facility. The instantaneous velocity field and surface heat transfer were determined simultaneously over a temporal sequence using a technique that combines traditional particle image velocimetry and thermochromic liquid crystal temperature measurements. The correlated results indicate that the highest surface heat transfer occurs in the trailing region of the spot where only the near-wall fluid is energetic. The results suggest that the "body" of the spot entrains, and subsequently recirculates, warm surface fluid within the spot, which reduces the effective heat transfer.

  4. Turbulent spot flow topology and mechanisms for surface heat transfer

    NASA Astrophysics Data System (ADS)

    Sabatino, D. R.; Smith, C. R.

    The properties of artificially initiated turbulent spots over a heated plate were investigated in a water channel. The instantaneous velocity field and surface Stanton number were simultaneously established using a technique that combines particle image velocimetry and thermochromic liquid crystal thermography. Several characteristics of a spot are found to be similar to those of a turbulent boundary layer. The spacing of the surface heat transfer streak patterns within the middle or of a turbulent spot are comparable to the low-speed streak spacing within a turbulent boundary layer. Additionally, the surface shear stress in the same region of a spot is also found to be comparable to a turbulent boundary layer. However, despite these similarities, the heat transfer within the spot body is found to be markedly less than the heat transfer for a turbulent boundary layer. In fact, the highest surface heat transfer occurs at the trailing or calmed region of a turbulent spot, regardless of maturity. Using a modified set of similarity coordinates, instantaneous two-dimensional streamlines suggest that turbulent spots entrain and subsequently recirculate warm surface fluid, thereby reducing the effective heat transfer within the majority of the spot. It is proposed that energetic vortices next to the wall, near the trailing edge of the spot body, are able to generate the highest surface heat transfer because they have the nearest access to cooler free-stream fluid.

  5. Acute infectious purpura fulminans due to probable spotted fever.

    PubMed

    Kundavaram, A; Francis, N R; Jude, A P J; Varghese, G N

    2014-01-01

    Purpura fulminans (PF) is associated with several infections, most notably with meningococcus, staphylococcus, and streptococcus infections. However, there are few reports of association of this entity with spotted fever from India. We report the case of a 55-year-old man who presented with fever, headache, and myalgia. On the seventh day of fever he developed nonblanching purple hemorrhagic purpura on the trunk and most prominently on the extremities consistent with purpura fulminans. Immunofluorescent assay confirmed the diagnosis of spotted fever. PF though common with rocky mountain spotted fever (RMSF) is rarely seen in association with Indian tick typhus, the usual cause of spotted fever in India.

  6. X-linked albinism-deafness syndrome and Waardenburg syndrome type II: A hypothesis

    SciTech Connect

    Zlotogora, J.

    1995-11-20

    Margolis reported on a large pedigree with a {open_quotes}new{close_quotes} X-linked syndrome of profound deafness and albinism (MIM 300700, albinism-deafness syndrome). The affected males presented with profound deafness and severe pigmentary abnormalities of the skin. At birth the skin appeared as almost albinotic except for areas of light pigmentation over the gluteal and scrotal areas, and thereafter pigmentation gradually increased over the body. Skin changes ultimately included areas of hypopigmentation and spots of hyperpigmentation. Some of the affected males also had blue irides, heterochromia, or segmental color iris changes. In carrier females, variable hearing impairment was documented without any pigmentary changes. 9 refs., 1 fig.

  7. Tourette Syndrome.

    ERIC Educational Resources Information Center

    Look, Kathy

    Tourette Syndrome has a history of being misdiagnosed or undiagnosed due to its unusual and complex symptoms. This paper describes: the symptoms of Tourette Syndrome; its etiology; age of onset; therapeutic methods, such as drug therapy, psychotherapy, diet control, and hypnosis; educational implications; and employment prospects. Several…

  8. Cardiorenal syndrome

    PubMed Central

    2009-01-01

    Kidney dysfunction in patients with heart failure and cardiovascular disorders in patients with chronic kidney disease are common. A recently proposed consensus definition of cardiorenal syndrome stresses the bidirectional nature of these heart-kidney interactions. The treatment of cardiorenal syndrome is challenging, however, promising new therapeutic options are currently being investigated in recent and ongoing clinical trials. PMID:20948701

  9. Down syndrome

    MedlinePlus

    ... Parents and caregivers should learn to help a person with Down syndrome deal with frustration. At the same time, it is important to encourage independence. Teen girls and women with Down syndrome are usually able to get pregnant. There is an increased risk of sexual abuse ...

  10. Turner Syndrome

    MedlinePlus

    ... opportunity to exchange ideas, develop coping strategies and locate resources. Peer groups for girls with Turner syndrome can help reinforce your daughter's self-esteem and provide her with a social network of people who understand her experience with Turner syndrome. References ...

  11. Turner syndrome

    MedlinePlus

    ... at birth is often smaller than average. A child with Turner syndrome is much shorter than children who are the ... Growth hormone may help a child with Turner syndrome grow taller. ... started when the girl is 12 or 13 years old. These help trigger ...

  12. Spotted black snake (Pseudechis guttatus) envenoming.

    PubMed

    Jansen, Melanie; McLeod, Monique; White, Julian; Isbister, Geoffrey K

    2007-01-01

    We report two cases of spotted black snake (Pseudechis guttatus) envenoming. One patient experienced localised burning pain around the bite and developed nausea, vomiting, diarrhoea, upper abdominal cramping and diaphoresis. He was treated with intravenous fluids and antiemetics, but no antivenom, and was discharged 23 hours after the bite. The second patient developed a severe headache, blurred vision and mild nausea, associated with severe pain and swelling of the bitten limb that took 4 days to resolve. No antivenom was given and the patient had no sequelae. Neither patient developed significant coagulopathy, myolysis or neuromuscular paralysis. Bites by this species appear to cause effects similar to those of the more common red-bellied black snake (P. porphyriacus).

  13. SPOT satellite mapping of Ice Stream B

    NASA Technical Reports Server (NTRS)

    Merry, Carolyn J.

    1993-01-01

    Numerous features of glaciological significance appear on two adjoining SPOT High Resolution Visible (HRV) images that cover the onset region of ice stream B. Many small-scale features, such as crevasses and drift plumes, have been previously observed in aerial photography. Subtle features, such as long flow traces that have not been mapped previously, are also clear in the satellite imagery. Newly discovered features include ladder-like runners and rungs within certain shear margins, flow traces that are parallel to ice flow, unusual crevasse patterns, and flow traces originating within shear margins. An objective of our work is to contribute to an understanding of the genesis of the features observed in satellite imagery. The genetic possibilities for flow traces, other lineations, bands of transverse crevasses, shear margins, mottles, and lumps and warps are described.

  14. Multiscale Simulation of Hot Spot Ignition

    NASA Astrophysics Data System (ADS)

    Fried, Laurence; Najjar, Fady; Howard, W. Michael; Manaa, M. Riad; Bastea, Sorin

    2011-06-01

    High explosive shock sensitivity is controlled by a combination of mechanical response, thermal properties, and chemical properties. How these properties interplay in realistic condensed energetic materials is not well understood. In this paper, we use a multiscale approach to achieve a realistic simulation of hot spot (void) ignition in a single crystal of the explosive 1,3,5-triamino-2,4,6-trinitrobenzene (TATB). The smallest length scale (<10 nm) of the multiscale model was treated quantum mechanically. We have conducted multiple simulations of the decomposition of the explosive TATB using density functional tight binding molecular dynamics (DFTB-MD). Nanoscale continuum simulations were performed of void ignition using the ALE3D hydrodynamic/thermal/chemical code. This work was performed under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344.

  15. Spot and Runway Departure Advisor (SARDA)

    NASA Technical Reports Server (NTRS)

    Jung, Yoon

    2016-01-01

    Spot and Runway Departure Advisor (SARDA) is a decision support tool to assist airline ramp controllers and ATC tower controllers to manage traffic on the airport surface to significantly improve efficiency and predictability in surface operations. The core function of the tool is the runway scheduler which generates an optimal solution for runway sequence and schedule of departure aircraft, which would minimize system delay and maximize runway throughput. The presentation also discusses the latest status of NASA's current surface research through a collaboration with an airline partner, where a tool is developed for airline ramp operators to assist departure pushback operations. The presentation describes the concept of the SARDA tool and results from human-in-the-loop simulations conducted in 2012 for Dallas-Ft. Worth International Airport and 2014 for Charlotte airport ramp tower.

  16. Syndromic craniosynostosis.

    PubMed

    Derderian, Christopher; Seaward, James

    2012-05-01

    Although most cases of craniosynostosis are nonsyndromic, craniosynostosis is known to occur in conjunction with other anomalies in well-defined patterns that make up clinically recognized syndromes. Patients with syndromic craniosynostoses are much more complicated to care for, requiring a multidisciplinary approach to address all of their needs effectively. This review describes the most common craniosynostosis syndromes, their characteristic features and syndrome-specific functional issues, and new modalities utilized in their management. General principles including skull development, the risk of developing increased intracranial pressure in craniosynostosis syndromes, and techniques to measure intracranial pressure are discussed. Evolving techniques of the established operative management of craniosynostosis are discussed together with more recent techniques including spring cranioplasty and posterior cranial vault distraction osteogenesis.

  17. Linburg syndrome

    PubMed Central

    Rennie, William R.J.; Muller, Hellmuth

    1998-01-01

    Objective To review the causes and demographics of Linburg syndrome. Design An illustrative case report and a demographic study. Setting Adult and pediatric orthopedic clinics at the Health Sciences Centre in Winnipeg. Patients One patient with Linburg syndrome and 200 patients and relatives presenting to adult and pediatric orthopedic clinics with conditions not involving their hands, wrists or forearms. Outcome measures The presence of the intertendinous anomaly and of carpal tunnel syndrome. Results Tendinous connection(s) between flexor pollicis longus and flexor digitorum profundus muscles were found in 20% of the study population. The anomaly was found in all age groups. No association was found between Linburg syndrome and the presence of carpal tunnel syndrome or previous injury to the hand or forearm. Conclusion Tendinous connection between flexor pollicis longus and flexor digitorum profundus muscles is a common anomaly that rarely causes clinical symptoms. PMID:9711164

  18. Demography of Northern Spotted Owls in southwestern Oregon

    USGS Publications Warehouse

    Zabel, Cynthia J.; Salmons, Susan E.; Forsman, Eric D.; Destefano, Stephen; Raphael, Martin G.; Gutierrez, R.J.

    1996-01-01

    Northern Spotted Owls (Strix occidentalis caurina) are associated with lower elevation, commercially valuable, late-successional coniferous forests in the Pacific Northwest. Meta-analyses of demographic parameters indicate that Northern Spotted Owl populations are declining throughout their range (Anderson and Burnham 1992, Burnham et al. this volume). Recent research has attempted to determine whether management activities have affected the viability of Spotted Owl populations, and results have led to development of conservation plans for the species (Dawson et al. 1987, Thomas et al. 1990, Murphy and Noon 1992, USDI 1992, Thomas et al. 1993b).In the Recovery Plan for the Northern Spotted Owl (USDI 1992b) threats to the species were identified as small population sizes, declining populations, limited amounts of habitat, continued loss and fragmentation of habitat, geographically isolated populations, and predation and competition from other avian species. Weather and fire are natural processes that also may affect reproductive success of Spotted Owls. Weather may be a factor in the high annual variability in fecundity of Spotted Owls, as has been suggested for other predatory bird species (Newton, 1979, 1986). However, these factors have not been addressed in previous studies of Spotted Owls.Our objectives were to estimate survival, fecundity, and annual rates of population change (l) for resident, territorial female Spotted Owls at two study areas in the coastal mountains of southwestern Oregon. We tested if the amount of rainfall was correlated with reproduction of Spotted Owls. While surveying for Spotted Owls, we documented the increased presence of Barred Owls (Strix varia), a potential competitor of Spotted Owls.

  19. Escobar syndrome mimicing congenital patellar syndrome.

    PubMed

    Ezirmik, Naci; Yildiz, Kadri; Can, Cahit Emre

    2012-08-01

    Multiple pterygium syndrome (MPS) is a syndrome that is characterized abnormal face, short length and skin pterygiums on some body legions (servical, antecubital, popliteal, interdigital and on neck). It is also called as Pterygium Colli syndrome, Escobar syndrome or Pterygium syndrome. Escobar (multyple pterygium) syndrome is a rare syndrome. Intrauterin growth reterdation, abnormal face, wide-spead pterygiums that resulted in joint contractures, ptosis, chryptoorchidism, patellar dysplasia and foot deformities are seen on this syndrome. Primarly autosomal resesive crossing are observed; also autosomal dominant and X-linked crossing. This case were presented as it has components of Escobar syndrome and Isolated Patellar Aplasia syndrome in same time.

  20. 7 CFR 28.422 - Strict Middling Spotted Color.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 2 2011-01-01 2011-01-01 false Strict Middling Spotted Color. 28.422 Section 28.422 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Color. Strict Middling Spotted Color is color which is within the range represented by a set of...