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1.
Environ Res ; 259: 119432, 2024 Oct 15.
Article in English | MEDLINE | ID: mdl-38944104

ABSTRACT

The Mediterranean Basin has experienced substantial land use changes as traditional agriculture decreased and population migrated from rural to urban areas, which have resulted in a large forest cover increase. The combination of Landsat time series, providing spectral information, with lidar, offering three-dimensional insights, has emerged as a viable option for the large-scale cartography of forest structural attributes across large time spans. Here we develop and test a comprehensive framework to map forest above ground biomass, canopy cover and forest height in two regions spanning the most representative biomes in the peninsular Spain, Mediterranean (Madrid region) and temperate (Basque Country). As reference, we used lidar-based direct estimates of stand height and forest canopy cover. The reference biomass and volume were predicted from lidar metrics. Landsat time series predictors included annual temporal profiles of band reflectance and vegetation indices for the 1985-2023 period. Additional predictor variables including synthetic aperture radar, disturbance history, topography and forest type were also evaluated to optimize forest structural attributes retrieval. The estimates were independently validated at two temporal scales, i) the year of model calibration and ii) the year of the second lidar survey. The final models used as predictor variables only Landsat based metrics and topographic information, as the available SAR time-series were relatively short (1991-2011) and disturbance information did not decrease the estimation error. Model accuracies were higher in the Mediterranean forests when compared to the temperate forests (R2 = 0.6-0.8 vs. 0.4-0.5). Between the first (1985-1989) and the last (2020-2023) decades of the monitoring period the average forest cover increased from 21 ± 2% to 32 ± 1%, mean height increased from 6.6 ± 0.43 m to 7.9 ± 0.18 m and the mean biomass from 31.9 ± 3.6 t ha-1 to 50.4 ± 1 t ha-1 for the Mediterranean forests. In temperate forests, the average canopy cover increased from 55 ± 4% to 59 ± 3%, mean height increased from 15.8 ± 0.77 m to 17.3 ± 0.21m, while the growing stock volume increased from 137.8 ± 8.2 to 151.5 ± 3.8 m3 ha-1. Our results suggest that multispectral data can be successfully linked with lidar to provide continuous information on forest height, cover, and biomass trends.


Subject(s)
Biomass , Environmental Monitoring , Forests , Spain , Environmental Monitoring/methods , Remote Sensing Technology , Trees/growth & development
2.
Chem Commun (Camb) ; 53(18): 2717-2720, 2017 Feb 28.
Article in English | MEDLINE | ID: mdl-28197588

ABSTRACT

Using dual mesoporous titania as a support, due to the presence of intrinsic Brönsted acid sites, the main approach to 4,6-dimethylbenzothiophene (46DMDBT) hydrodesulfurization (HDS) becomes the direct desulfurization (DDS) route through isomerization and dismutation reactions, instead of the hydrogenation (HYD) pathway usually observed with a conventional promoted (by Ni or Co) MoS2/Al2O3 catalyst.

3.
Pediatr. aten. prim ; 16(64): e147-e150, oct.-dic. 2014. tab
Article in Spanish | IBECS | ID: ibc-131215

ABSTRACT

La artritis idiopática juvenil, a pesar de su baja incidencia, es la enfermedad reumática inflamatoria crónica más frecuente en la infancia. Ahora disponemos de criterios precisos y sencillos para diagnosticarla en Atención Primaria, ya que son criterios fundamentalmente clínicos. El diagnóstico de mono- o poliartritis, en ausencia de síntomas infecciosos generales y con radiografías normales, debe hacernos sospecharla en pacientes menores de 16 años. También nos hará pensar en ella su persistencia durante más de seis semanas. Las distintas formas clínicas serán diagnosticadas por el reumatólogo, pero la sospecha de la misma se debe establecer en Atención Primaria. El objetivo es conocer el tratamiento inicial para controlar el dolor y la inflamación, mantener la función y favorecer un crecimiento normal. No hay que olvidar las posibles complicaciones oculares. Enviar a Oftalmología en el momento del diagnóstico y realizar revisiones anuales así como al presentar cualquier síntoma que afecte a la cámara anterior (AU)


The Juvenile idiophatic arthritis in spite of his low frequency, is the most frequent rheumatic inflammatory chronic disease in the infancy. Now we have precise and simple criteria to diagnose her in Primary care since they are criteria fundamentally clinical and spread on the time. The diagnosis of monkey or polyarthritis, in absence of infectious general symptoms and with normal X-ray photographies, must make us suspect her in 16-year-old minor patients. Also it will make us think about her his persistence of more than 6 weeks. The different clinical forms will be diagnosed by the reumatólogo. But the suspicion of the same one must establish it the doctor of Primary care or the pediatrician of the Center of Health. The aim is to know the initial treatment that controls the pain and the inflammation, to support the function and to favor a normal growth. It is not necessary to forget the possible ocular complications. To send to ophthalmology in the moment of the diagnosis and to realize annual reviews as well as on having presented any symptom that concerns the previous chamber (AU)


Subject(s)
Humans , Female , Child , Arthritis, Juvenile/complications , Arthritis, Juvenile/diagnosis , Arthritis, Juvenile/drug therapy , Arthritis, Rheumatoid/complications , Arthritis, Rheumatoid/drug therapy , Pain Management/methods , Primary Health Care/methods , Primary Health Care/trends , Primary Health Care
4.
Plant Dis ; 97(7): 991, 2013 Jul.
Article in English | MEDLINE | ID: mdl-30722571

ABSTRACT

Cork oaks (Quercus suber L.) are key tree species at Doñana Biological Reserve (DBR), Huelva, Spain. Sampling was conducted on a total of 13 trees exhibiting symptoms of decline (foliar wilting and defoliation, branch dieback, and root necrosis). In 2008. Phytophthora cinnamomi was isolated from feeder roots of one tree and Pythium spiculum from two additional oaks. In 2011, both pathogens were isolated from six oaks, only P. cinnamomi from three oaks, and only Py. spiculum from one oak. This expansion was associated with high winter rainfall levels since 2009 that led to long periods of soil flooding. While P. cinnamomi is well known to cause a root disease on Q. suber (2), P. spiculum is a newly described species isolated from Quercus, Vitis, Prunus, Castanea, and Celtis species, but its pathogenicity was demonstrated only on Q. ilex (syn. Q. rotundifolia) (1). Pathogenicity tests were conducted on 4-year-old Q. suber plants. Inocula consisted of two isolates of Py. spiculum from DBR (DO8 and DO36 from Q. suber). For comparison with these, three isolates previously tested on Q. ilex (1) were included: two isolates of Py. spiculum, PA54 (from Q. suber) and PE156 (from Q. ilex); and one isolate of P. cinnamomi, PE90 (from Q. ilex). All these isolates came from the Andalucía region, stored at the oomycete collection of the University of Córdoba, and showed a 99 to 100% homology with their expected ITS sequences in GenBank (DQ196131 for Py. spiculum and AY943301 for P. cinnamomi). Inoculum was prepared by shaking and mixing propagule-bearing mycelium produced in carrot broth petri dishes (20°C, 4 weeks) in sterile water, to produce a concentration of 3 × 104 oospores × ml-1 (Py. spiculum) or 3 × 104 chlamydospores × ml-1 (P. cinnamomi). One hundred milliliters of inoculum was applied to each root (1). There were 10 inoculated plants per isolate and 10 non-inoculated control plants. All plants were waterlogged 2 days per week to favor root infection and maintained in an acclimatised greenhouse (12-28°C). Three months later, the inoculated plants showed symptoms of root necrosis that resulted in foliar wilting followed occasionally by defoliation. Control plants did not develop foliar symptoms nor root necrosis. Root damage severity assessed on a 0 to 4 scale (3) exhibited significant differences (P < 0.05) in relation to the control plants for all the isolates tested, with isolate PE90 (P. cinnamomi) and isolates PA54, DO8, and DO36 (P. spiculum) all averaging a root necrosis value of 2.5. Isolate PE156 of P. spiculum produced values of root necrosis (1.6 in average) significantly lower (P < 0.05) than the rest. This isolate belongs to the low virulence group of P. spiculum described on Q. ilex (1). The inoculated oomycete was always reisolated from necrotic roots and never from roots of control plants. To the best of our knowledge, this is the first report of P. spiculum as the cause of root rot of Q. suber. References: (1) Romero et al. J. Phytopathol. 155:289, 2007. (2) Sánchez et al. For. Pathol. 32:5, 2002. (3) Sánchez et al. For. Pathol. 35:115, 2005.

5.
Neuroscience ; 207: 182-97, 2012 Apr 05.
Article in English | MEDLINE | ID: mdl-22300980

ABSTRACT

Cocaine's multiple pharmacological substrates are ubiquitously present in the peripheral and central nervous system. Thus, upon its administration, cocaine acts in the periphery before directly acting in the brain. We determined whether cocaine alters ventral tegmental area (VTA) neuronal activity via its peripheral actions. In urethane-anesthetized rats, we recorded VTA neuron's responses to intravenous injections of two cocaine analogs: cocaine-hydrochloride (HCl, 0.25 mg/kg), which readily cross the blood-brain barrier (BBB), and cocaine-methiodide (MI, 0.33 mg/kg), which does not cross the BBB. Both cocaine analogs produced sustained changes in discharge rates that began 5 s after the initiation of a 10-s drug infusion. Within the first 90 s post-injection, the magnitudes of neuronal responsiveness of both cocaine analogs were comparable, but later the effects of cocaine-HCl were stronger and persisted longer than those of cocaine-MI. The proportion of neurons responsive to cocaine-HCl was twice that of cocaine-MI (74% and 35%, respectively). Both analogs also differed in their response onsets. Cocaine-MI rarely evoked responses after 1 min, whereas cocaine-HCl continued to evoke responses within 3 min post-injection. VTA neurons were either excited or inhibited by both cocaine analogs. Most units responsive to cocaine-MI, regardless of whether they were excited or inhibited, had electrophysiological characteristics of putative dopamine (DA) neurons. Units inhibited by cocaine-HCl also had characteristics of DA neurons, whereas excited neurons had widely varying action potential durations and discharge rates. Cocaine-MI and cocaine-HCl each produced changes in VTA neuron activity under full DA receptor blockade. However, the duration of inhibition was shortened and the number of excitations increased, and they occurred with an earlier onset during DA receptor blockade. These findings indicate that cocaine acts peripherally with a short latency and alters the activity of VTA neurons before its well-known direct actions in the brain.


Subject(s)
Cocaine-Related Disorders/physiopathology , Cocaine/pharmacokinetics , Dopaminergic Neurons/drug effects , Peripheral Nervous System/drug effects , Ventral Tegmental Area/drug effects , Animals , Cocaine/analogs & derivatives , Disease Models, Animal , Dopamine Uptake Inhibitors/pharmacokinetics , Dopaminergic Neurons/physiology , Injections, Intravenous/methods , Male , Peripheral Nervous System/physiology , Rats , Rats, Long-Evans , Ventral Tegmental Area/physiology
6.
Med Eng Phys ; 31(7): 775-83, 2009 Sep.
Article in English | MEDLINE | ID: mdl-19297233

ABSTRACT

The detailed quantitative characterization of soft-tissue in-growth into highly porous artificial implants is critical to understanding the biophysical processes that will lead to the best structural scaffolding construct. Previous studies have performed mechanical peel tests and mostly qualitative histological analyses of soft-tissue. The goal of this paper is to report the results obtained from applying two image analysis algorithms to quantify the morphological structure found in histological images of stained soft-tissue in-growth into alumina ceramic foam metal implants using a canine model. Three different pore sizes were used and three different post-operative time points were considered. Using the 2D Wavelet Transform Modulus Maxima method and 2D Fourier Transform analysis, a strong anisotropic signature (directional preference) is detected in early (4-week) histological samples. The direction of preference is towards the center of the implants. The strength of the anisotropy at later time points (8 and 16 weeks) becomes gradually weaker. Our interpretation is that after a short period of time, the main tissue growth activity has been concentrated on filling the artificial implant by growing towards its center. The weaker anisotropic signature found at later time points is interpreted as the tissue growth activity strengthening its structure by growing in more random directions.


Subject(s)
Aluminum Oxide/chemistry , Image Processing, Computer-Assisted/methods , Metals/chemistry , Tissue Scaffolds/chemistry , Animals , Anisotropy , Dogs , Gene Expression Regulation , Intercellular Signaling Peptides and Proteins/metabolism , Models, Animal , Porosity , Plastic Surgery Procedures , Surface Properties , Time Factors , Tissue Engineering
7.
Rheumatology (Oxford) ; 46(3): 398-402, 2007 Mar.
Article in English | MEDLINE | ID: mdl-17085771

ABSTRACT

OBJECTIVE: Localized scleroderma causes thickening of the skin due to excessive collagen deposition. This condition has clinical and histopathological similarities to chronic graft-vs-host disease. We wanted to identify whether chimeric cells are present in the affected tissue in localized scleroderma and to further investigate the role of chimerism by immunophenotyping the chimeric cells. We hypothesize that the presence of chimerism and immunotypic chimeric cells will lend to an understanding of the pathogenesis of localized scleroderma and possible mechanisms by which chimeric cells participate in autoimmunity. METHODS: We studied skin biopsies from 18 localized scleroderma patients and compared them with concurrent biopsies from unaffected skin in a subset of patients. Skin biopsies from morphoea and linear scleroderma patients were analysed for the presence of chimeric cells using male-female (X, Y) differences. Cell surface markers (CD4, CD8, CD19/20, CD68, S100, CD14 and CD56) were determined for cell phenotyping of chimeric cells. RESULTS: Overall, the affected tissue contained a greater number of lymphocytic inflammatory cells. In the affected tissue, 38% of the total chimeric cells were CD68+ (dendritic cell, monocyte and macrophage marker), 29% Langerin/S100+ (dendritic cell marker), 26% CD8+ (cytotoxic T-lymphocyte marker), 20% CD19/20+ (B-lymphocyte marker), 14% CD4+ (T-helper lymphocyte) and 0% CD56+ (natural killer cell marker). CONCLUSIONS: We report that not only are chimeric cells present in affected localized scleroderma lesions but they also are more likely to be dendritic cells and B lymphocytes suggesting a role in the pathogenesis of localized scleroderma.


Subject(s)
Autoimmune Diseases/immunology , Chimera/immunology , Scleroderma, Localized/immunology , Adolescent , Adult , Aged , B-Lymphocytes/immunology , Child , Dendritic Cells/immunology , Female , Humans , Immunophenotyping , Male , Middle Aged , Skin/immunology
8.
Sex Transm Infect ; 82(4): 311-6, 2006 Aug.
Article in English | MEDLINE | ID: mdl-16877581

ABSTRACT

OBJECTIVE: Assessment of HIV prevalence and associated risk behaviours among female commercial sex workers (FCSW) across major cities in South America. METHODS: Seroepidemiological, cross sectional studies of 13 600 FCSW were conducted in nine countries of South America during the years 1999-2002. Participants were recruited in brothels, massage parlours, hotels, and streets where anonymous questionnaires and blood samples were collected. HIV infection was determined by enzyme linked immunosorbent assay (ELISA) screening and western blot confirmatory tests. RESULTS: The overall HIV seroprevalence was 1.2% (range 0.0%-4.5%). The highest HIV seroprevalences were reported in Argentina (4.5%) and Paraguay (2.6%); no HIV infected FCSW were detected in Venezuela and Chile. Consistent predictors of HIV seropositivity were: (1) a previous history of sexually transmitted infections (STI, AORs = 3.8-8.3), and (2) 10 years or more in commercial sex work (AORs = 2.2-24.8). In addition, multiple (> or =3) sexual contacts (AOR = 5.0), sex with foreigners (AOR = 6.9), use of illegal drugs (AOR = 3.2), and marijuana use (AOR = 8.2) were associated with HIV seropositivity in Southern Cone countries. CONCLUSIONS: Consistently low HIV seroprevalences were detected among FCSW in South America, particularly in the Andean region. Predictors of HIV infection across the continent were STI and length of commercial sex work; however, use of illegal drugs, especially marijuana, and sexual contacts with foreigners were also found to be associated risk factors in the Southern Cone region. Interventions for the control of HIV and other STI need to be region and country specific; drug use appears to have an ever increasing role in the spread of HIV among heterosexually active populations.


Subject(s)
HIV Infections/epidemiology , HIV Seroprevalence , HIV-1 , Sex Work/statistics & numerical data , Adult , Blotting, Western , Enzyme-Linked Immunosorbent Assay , Epidemiologic Methods , Female , Humans , Risk Factors , South America/epidemiology
9.
Ann N Y Acad Sci ; 965: 157-71, 2002 Jun.
Article in English | MEDLINE | ID: mdl-12105092

ABSTRACT

The repeated use of psychostimulants in humans has been associated with progressive enhancement of anxiety, panic attacks, and eventually paranoid psychosis. The appearance of such behaviors has been termed behavioral sensitization, which forms part of the basic pathological mechanisms involved in drug addiction. Psychostimulants act via a circuit involving the ventral tegmental area (VTA), prefrontal cortex (PFC), and nucleus accumbens. The PFC sends glutamatergic projections that activate dopaminergic neurons in the VTA. These projections provide an extremely important excitatory drive necessary for the development of sensitization. The effects of cocaine administration on the response of dopaminergic VTA cells to activation of the PFC have not been reported. Here the effects of acute cocaine administration on VTA cell response to PFC stimulation are examined. Statistical analysis of the changes in spontaneous activity and evoked response revealed a significant decrease in spontaneous activity at 1.0 mg/kg i.v. after cocaine treatment compared to baseline levels. The net effect was an increase in signal-to-noise ratio. Treatment with MK-801 at a dose of 2 mg/kg showed that the excitatory response was, at least partially, NMDA-mediated. Prazosin pretreatment (0.5 mg/kg i.p.) did not prevent a significant decrease in spontaneous activity brought about by cocaine (15 mg/kg, i.p.). Nonetheless, prazosin alone induced a significant decrease in the response to PFC stimulation when compared to baseline. In addition, iontophoretic application of norepinephrine (NE) onto VTA cells revealed that NE potentiated (19.2%), enhanced (26.9%), or suppressed (46.2%) the glutamate-evoked response in VTA cells. The results suggest that a possible role of cocaine in the process of sensitization might be to amplify the PFC-induced excitation at the VTA. Since the iontophoretic release of NE in almost half of the sampled cells produced similar effects to those of cocaine it may suggest a possible NE-mediated mechanism for cocaine actions.


Subject(s)
Cocaine/pharmacology , Neurons/physiology , Prefrontal Cortex/physiology , Ventral Tegmental Area/physiology , Animals , Cocaine/administration & dosage , Electric Stimulation , Glutamic Acid/pharmacology , Injections, Intravenous , Male , Neurons/drug effects , Norepinephrine/pharmacology , Rats , Rats, Sprague-Dawley , Ventral Tegmental Area/drug effects
10.
Article in Spanish | LILACS | ID: lil-430667

ABSTRACT

Entre los virus conocidos, uno de los cuales se le ha dedicado una extensiva atenci¢n es la epidemiolog¡a molecular del virus de la inmunodeficiencia humana (VIH). Más recientemente, el intéres se ha expandido esencialmente al origen y evoluci¢n de su estructura genética, mecanismos patogénicos y respuestas inmune. Este art¡culo describe recientes trabajos para la comprensi¢n del origen y evoluci¢n del VIH


Subject(s)
Biological Evolution , History , HIV , RNA , Medicine , Venezuela
11.
J Antimicrob Chemother ; 45(5): 645-9, 2000 May.
Article in English | MEDLINE | ID: mdl-10797087

ABSTRACT

Minimum inhibitory concentrations (MICs) of amphotericin B, 5-flucytosine, fluconazole, itraconazole and ketoconazole were determined against 42 clinical isolates of Cryptococcus neoformans var. neoformans using the Alamar YeastOne colorimetric method and the NCCLS reference microdilution method. No strains with resistance to amphotericin B, itraconazole or ketoconazole were detected with either method. Using the reference method, the MICs of fluconazole were >/= 64 mg/L, whereas using the colorimetric method all MICs were >/=16 mg/L. The MIC values of 5-flucytosine were also higher using the reference method (8-16 mg/L for 32% of isolates) compared with the colorimetric method. The percentage of agreement between the methods, using a difference of two dilutions, was 70.7% for itraconazole, 73.2% for amphotericin B, 80% for fluconazole, 88% for 5-flucytosine and 95% for ketoconazole. Overall, we conclude that for fluconazole and 5-flucytosine, in a low but not insignificant number of isolates, results with the two methods are discordant, some isolates being found sensitive with the colorimetric test, but resistant with the reference method.


Subject(s)
Antifungal Agents/pharmacology , Cryptococcus neoformans/drug effects , Colorimetry , Cryptococcosis/microbiology , Cryptococcus neoformans/isolation & purification , Drug Resistance, Microbial , Humans , Microbial Sensitivity Tests/methods , Reference Standards , Reproducibility of Results
12.
Adm Policy Ment Health ; 28(2): 91-106, 2000 Nov.
Article in English | MEDLINE | ID: mdl-11194126

ABSTRACT

The authors utilized numerous documents created by advisory groups, expert panels and multicultural focus groups to develop performance measures for assessing the cultural competency of mental health systems. Competency was measured within three levels of organizational structure: administrative, provider network, and individual caregiver. Indicators, measures and data sources for needs assessment, information exchange, services, human resources, plans and policies, and outcomes were identified. Procedures for selection and implementation of the most critical measures are suggested. The products of this project are broadly applicable to the concerns of all cultural groups.


Subject(s)
Cultural Diversity , Management Audit , Mental Health Services/organization & administration , Professional Competence , Quality Indicators, Health Care , Focus Groups , Humans , Mental Health Services/standards , Minority Groups , Needs Assessment , New York , Organizational Policy , Program Evaluation/methods , Public Health Administration/standards
13.
J Virol ; 70(2): 985-91, 1996 Feb.
Article in English | MEDLINE | ID: mdl-8551639

ABSTRACT

Replication of the rotavirus genome involves two steps: (i) transcription and extrusion of transcripts and (ii) minus-strand RNA synthesis in viral complexes containing plus-strand RNA. In this study, we showed evidence for the importance of the viral nonstructural protein of rotavirus, NSP2, in the replication of viral RNAs. RNA-binding properties of NSP2 were tested by UV cross-linking in vivo (in rotavirus-infected MA104 cells and recombinant baculovirus-expressing NSP2-infected Sf9 cells). In rotavirus-infected cells, NSP2 is bound to the 11 double-stranded RNA genomic segments of rotavirus. Quantitative analysis (using hydrolysis by RNase A) is consistent with NSP2 being directly bound to partially replicated viral RNA. Using various monoclonal antibodies and specific antisera against the structural (VP1, VP2, and VP6) and nonstructural (NSP1, NSP2, NSP3, and NSP5) proteins, we developed a solid-phase assay for the viral replicase. In this test, we recovered a viral RNA-protein complex with replicase activity only with a monoclonal antibody directed against NSP2. Our results indicated that these viral complexes contain the structural proteins VP1, VP2, and VP6 and the nonstructural protein NSP2. Our results show that NSP2 is closely associated in vivo with the viral replicase.


Subject(s)
RNA-Binding Proteins/metabolism , RNA-Dependent RNA Polymerase/isolation & purification , Rotavirus/enzymology , Viral Nonstructural Proteins/metabolism , Animals , Antibodies, Monoclonal/immunology , Cattle , Cell Line , Cross-Linking Reagents , Photochemistry , RNA, Viral/metabolism , RNA-Binding Proteins/chemistry , RNA-Binding Proteins/immunology , RNA-Dependent RNA Polymerase/chemistry , RNA-Dependent RNA Polymerase/metabolism , Rotavirus/physiology , Templates, Genetic , Viral Nonstructural Proteins/chemistry , Viral Nonstructural Proteins/immunology , Virus Replication
14.
Arch Virol Suppl ; 12: 29-35, 1996.
Article in English | MEDLINE | ID: mdl-9015099

ABSTRACT

The genomes of viruses in the family Reoviridae consist of segmented double-stranded RNA. There are 10 to 12 segments depending on the genus. The 5' ends and the 3' ends of the RNAs present conserved motifs for each virus genus. These conserved motifs have been hypothesized to play a role in genomic segment assortment during virus morphogenesis. Using a set of monoclonal antibodies we have tried to identify rotaviral proteins that bind to RNA during infection in cell culture. This methodology takes advantage of being able to label RNA in vitro to high specific activity and also of solid phase processing of RNA-protein complexes. After cross-linking the RNA to protein in infected cells, protein-RNA complexes are precipitated with a specific MAb; then, the RNA in the complex is labeled in vitro and the protein or nucleic acid moieties are analyzed by usual protocols. This paper describes results using an anti NSP3 MAb. In infected cells, we have shown that NSP3 binds to the eleven messenger RNAs, and that a sequence from nucleotides 8 to 15 is protected from digestion with RNAse T1 by NSP3 in the RNA-protein complex. The availability of recombinant protein NSP3 expressed in the baculovirus-insect cell system has allowed the sequence specificity of NSP3 to be studied in vitro. The minimal sequence recognized by NSP3 is GACC. The role of NSP3 in rotavirus replication is discussed based on these results and by comparison with other RNA-binding proteins of members of the Reoviridae family.


Subject(s)
RNA, Viral/metabolism , Rotavirus/metabolism , Viral Nonstructural Proteins/chemistry , Viral Nonstructural Proteins/metabolism , Animals , Cell Line , Consensus Sequence , Humans , Protein Conformation , RNA, Messenger/metabolism , RNA, Viral/analysis , Recombinant Fusion Proteins/chemistry , Recombinant Fusion Proteins/metabolism , Rotavirus/chemistry , Rotavirus/genetics , Sequence Analysis, RNA , Spodoptera/cytology , Structure-Activity Relationship
15.
J Virol ; 67(6): 3159-65, 1993 Jun.
Article in English | MEDLINE | ID: mdl-8388495

ABSTRACT

Interaction between viral proteins and RNAs has been studied in rotavirus-infected cells. The use of UV cross-linking followed by immunoprecipitation and labeling with T4 polynucleotide kinase allowed us to detect interactions between RNA and nonstructural viral proteins. The RNAs linked to the nonstructural protein NSP3 have been identified as rotavirus mRNAs, and the sequences of the RNase T1-protected fragments have been established. These sequences correspond to the 3' end sequence common to all rotavirus group A genes. We also show that the last 3' nucleotide is cross-linked to the protein and that monomeric and multimeric forms of NSP3 are bound to rotavirus mRNA. The role of NSP3 in rotavirus replication is discussed in the light of our results and by comparison with other RNA-binding proteins of members of the Reoviridae family.


Subject(s)
RNA, Messenger/genetics , RNA, Viral/genetics , Rotavirus/genetics , Viral Nonstructural Proteins/metabolism , Base Sequence , Cells, Cultured , Consensus Sequence , Humans , Molecular Sequence Data , Protein Conformation , RNA, Messenger/metabolism , RNA, Viral/metabolism , Rotavirus/growth & development , Sequence Analysis, RNA
16.
Arch Virol ; 133(1-2): 85-95, 1993.
Article in English | MEDLINE | ID: mdl-8240020

ABSTRACT

Studies on rotavirus non-structural proteins have been hampered in the past by difficulties in obtaining monospecific reagents. To make such reagents available, we have expressed in the baculovirus system NSP2 and NSP3 (formerly called NS35 and NS34, respectively) of the bovine rotavirus RF and produced hybridomas against these proteins. Full-length DNA copies of RNA segments 7 (coding for NSP3) and 8 (coding for NSP2) of the virus strain RF were cloned and sequenced. Each cDNA was inserted in the transfer vector pVL941 and used to transfect Spodoptera frugiperda cells (Sf9). Recombinant baculoviruses encoding these proteins were obtained. Infection of Sf9 cells with these recombinant viruses resulted in a high level of expression of NSP2 and NSP3 (range of 1 microgram per 10(6) cells). Monoclonal antibodies (MAbs) were elicited by immunization of BALB/c mice with adjuvented, unpurified recombinant proteins in the rear foot pads. Fusion was performed using lymphocytes from popliteal lymph nodes with SP2/O-Ag14 myeloma line. Screening was by differential indirect immunofluorescent staining on monolayers of Sf9 cells infected with each recombinant virus. Two MAbs proved to be reactive against NSP3 and a single one against NSP2. They showed high specificity by immunofluorescence, immunoprecipitation and Western blot. The isotype of these MAbs was IgG1. Oligomeric forms of NSP3 and NSP2 proteins were detected and the existence of intra-chain disulfide bridge in NSP2 protein was suggested. The levels of synthesis and cellular localization of NSP3 and NSP2 proteins were different as shown by immunoprecipitation and immunofluorescence.


Subject(s)
Antibodies, Monoclonal/immunology , Rotavirus/genetics , Viral Nonstructural Proteins/genetics , Animals , Antibodies, Monoclonal/biosynthesis , Baculoviridae , Base Sequence , Cattle , Cell Line , Cloning, Molecular , DNA, Viral , Electrophoresis, Polyacrylamide Gel , Fluorescent Antibody Technique , Genes, Viral , Humans , Hybridomas , Molecular Sequence Data , Moths , Precipitin Tests , Recombinant Proteins , Viral Nonstructural Proteins/biosynthesis , Viral Nonstructural Proteins/immunology
17.
Virology ; 190(1): 68-83, 1992 Sep.
Article in English | MEDLINE | ID: mdl-1326821

ABSTRACT

Intermolecular interactions between polypeptide chains often play essential roles in such biological phenomena as replication, transcription, translation, transport, ligand binding, and assembly. We have initiated studies of the functions of the rotavirus SA114F gene 7 product by sequence analysis and expression in insect cells. This nonstructural protein, NS34, is a slightly acidic protein, and its secondary structure is predicted to be 78% alpha-helix, with several heptad repeats of hydrophobic amino acids being present in its carboxy half. NS34 was found in oligomers when analyzed in insect cells, in SA11-infected MA104 cells, and in cell-free translation reactions. Investigation of the multiple electrophoretically distinct forms of NS34 showed they were all composed of homooligomers. Deletion mutants constructed and tested for oligomerization showed that the carboxy terminus of the protein, containing the predicted heptad repeats, was responsible for oligomerization. A basic region present in NS34 of group A rotaviruses, found to be 40% conserved in NS34 of group C rotavirus, is a candidate for a functional domain of this protein. NS34, which was found to be associated with the cytoskeleton fraction of cells, also interacts with viral RNA. These results make it likely that NS34 plays a central role in the replication and assembly of genomic RNA structures.


Subject(s)
Capsid/chemistry , RNA-Binding Proteins/chemistry , Rotavirus/genetics , Viral Core Proteins/chemistry , Amino Acid Sequence , Animals , Capsid/genetics , Capsid/metabolism , Cell Line , Cloning, Molecular , Electrophoresis, Polyacrylamide Gel , Fluorescent Antibody Technique , Genes, Viral , Molecular Sequence Data , Protein Biosynthesis , RNA, Viral/metabolism , RNA-Binding Proteins/genetics , RNA-Binding Proteins/metabolism , Repetitive Sequences, Nucleic Acid , Rotavirus/physiology , Sequence Alignment , Viral Core Proteins/genetics , Viral Core Proteins/metabolism , Viral Nonstructural Proteins
18.
Clin Exp Rheumatol ; 7(3): 229-36, 1989.
Article in English | MEDLINE | ID: mdl-2667829

ABSTRACT

We report two Caucasian families with systemic sclerosis and other connective tissue and immunological disorders, including rheumatoid arthritis, discoid lupus erythematosus, psoriasis, psoriatic arthritis, ankylosing spondylitis, ulcerative colitis, asthma, Sjögren's syndrome, Raynaud's phenomenon and thyroid disease. In one of these families, two sisters are affected with systemic sclerosis. Clinical, serological, and HLA haplotype results are reported, along with a review of the medical literature on familial occurrence of systemic sclerosis.


Subject(s)
Arthritis, Rheumatoid/genetics , HLA Antigens/genetics , Immune System Diseases/genetics , Scleroderma, Systemic/genetics , Aged , Arthritis, Rheumatoid/immunology , Humans , Immune System Diseases/immunology , Male , Middle Aged , Pedigree , Phenotype , Scleroderma, Systemic/immunology
19.
Int J Addict ; 20(4): 643-50, 1985 Apr.
Article in English | MEDLINE | ID: mdl-4030178

ABSTRACT

A retrospective study of 781 alcoholics detoxified at two treatment centers suggested that magnesium sulfate was significant in preventing seizures and that benzodiazepines were essential in minimizing other complications. Future investigations should determine the most effective mineral dosage levels for alcohol detoxification.


Subject(s)
Alcohol Withdrawal Delirium/prevention & control , Alcoholism/drug therapy , Magnesium Sulfate/therapeutic use , Psychoses, Alcoholic/prevention & control , Benzodiazepines/therapeutic use , Ethanol/adverse effects , Humans , Retrospective Studies , Substance Withdrawal Syndrome/drug therapy
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