RESUMO
BACKGROUND: Spinal anaesthesia, the most common form of anaesthesia for caesarean section, leads to sympathetic blockade and profound maternal hypotension resulting in adverse maternal and neonatal outcomes. Hypotension, nausea and vomiting remain common but until the publication of the National Institute of Health and Care Excellence (NICE) 2021 guidance, no national guideline existed on how best to manage maternal hypotension following spinal anaesthesia for caesarean section. A 2017 international consensus statement recommended prophylactic vasopressor administration to maintain a systolic blood pressure of >90% of an accurate pre-spinal value, and to avoid a drop to <80% of this value. This survey aimed to assess regional adherence to these recommendations, the presence of local guidelines for management of hypotension during caesarean section under spinal anaesthesia, and the individual clinician's treatment thresholds for maternal hypotension and tachycardia. METHODS: The West Midlands Trainee-led Research in Anaesthesia and Intensive Care Network co-ordinated surveys of obstetric anaesthetic departments and consultant obstetric anaesthetists across 11 National Health Service Trusts in the Midlands, England. RESULTS: One-hundred-and-two consultant obstetric anaesthetists returned the survey and 73% of sites had a policy for vasopressor use; 91% used phenylephrine as the first-line drug but a wide range of recommended delivery methods was noted and target blood pressure was only listed in 50% of policies. Significant variation existed in both vasopressor delivery methods and target blood pressures. CONCLUSIONS: Although NICE has since recommended prophylactic phenylephrine infusion and a target blood pressure, the previous international consensus statement was not adhered to routinely.
Assuntos
Anestesia Obstétrica , Raquianestesia , Cesárea , Hipotensão , Vasoconstritores , Humanos , Feminino , Gravidez , Adulto , Hipotensão/etiologia , Raquianestesia/efeitos adversos , Anestesia Obstétrica/efeitos adversos , Reino Unido , Inquéritos e Questionários , Vasoconstritores/administração & dosagem , Vasoconstritores/efeitos adversosRESUMO
Curative intervention is possible if colorectal cancer is identified early, underscoring the need to detect the earliest stages of malignant transformation. A candidate biomarker is the expanded proliferative zone observed in crypts before adenoma formation, also found in irradiated crypts. However, the underlying driving mechanism for this is not known. Wnt signaling is a key regulator of proliferation, and elevated Wnt signaling is implicated in cancer. Nonetheless, how cells differentiate Wnt signals of varying strengths is not understood. We use computational modeling to compare alternative hypotheses about how Wnt signaling and contact inhibition affect proliferation. Direct comparison of simulations with published experimental data revealed that the model that best reproduces proliferation patterns in normal crypts stipulates that proliferative fate and cell cycle duration are set by the Wnt stimulus experienced at birth. The model also showed that the broadened proliferation zone induced by tumorigenic radiation can be attributed to cells responding to lower Wnt concentrations and dividing at smaller volumes. Application of the model to data from irradiated crypts after an extended recovery period permitted deductions about the extent of the initial insult. Application of computational modeling to experimental data revealed how mechanisms that control cell dynamics are altered at the earliest stages of carcinogenesis.
Assuntos
Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/radioterapia , Inibição de Contato/efeitos da radiação , Via de Sinalização Wnt/efeitos da radiação , Animais , Divisão Celular/efeitos da radiação , Proliferação de Células/fisiologia , Proliferação de Células/efeitos da radiação , Transformação Celular Neoplásica , Simulação por Computador , Regulação Neoplásica da Expressão Gênica , Humanos , Mucosa Intestinal/metabolismo , Masculino , Camundongos , Proteínas Wnt/metabolismoRESUMO
The complete nucleotide sequence of the RNA genome segment coding for the outer capsid protein, VP5, of the United States prototypic strain of bluetongue virus (BTV) serotype 11 was determined from two overlapping cDNA clones. The genome segment was found to be 1638 nucleotides in length with a single open reading frame coding for a 526 amino acid protein of MW 59,278 and having a net charge of -4.0 at neutral pH. Comparisons of the predicted amino acid sequence of VP5 of BTV 11 with those of the United States serotypes 2, 10, and 13 and two isolates of BTV 1 from Australia and South Africa confirmed earlier reports that VP5 is a conserved protein with no clear regions of variability. A computer generated consensus sequence suggested VP5 of BTV 2 to be representative of the average VP5 sequences reported thus far.
Assuntos
Vírus Bluetongue/genética , Capsídeo/genética , Sequência de Aminoácidos , Sequência de Bases , Vírus Bluetongue/imunologia , Sequência Consenso , Dados de Sequência Molecular , Fases de Leitura Aberta , RNA Viral/genética , Alinhamento de Sequência , SorotipagemRESUMO
A liquid phase hybridization format has been adapted for the identification of bluetongue virus (BTV) with respect to serogroup and serotype. In this paper we present experiments which demonstrate the identification of bluetongue virus with respect to both serogroup and serotype. BTV serogroup-specific and BTV serotype 17-specific single-stranded RNA probes labeled with 35S-CTP were used to identify BTV viral nucleic acids in infected cell culture lysates. Cell lysates were prepared for testing by rapid solubilization in a guanidine isothiocyanate/EDTA/dithiothreitol medium. Hybridizations were done in 12.5 microliter reaction volumes for 2 h or overnight using nanogram quantities of probe. An RNAse solution was added subsequent to hybridization and incubated for 30 min. TCA-precipitable counts were collected and quantitated by scintillation counting. Specific hybridization signals were obtained in as little as 3 h in assays consisting of uninfected controls, cells infected with different BTV serotypes, and cells infected with a strain of epizootic hemorrhagic disease of deer (EHDV-1). By using asymmetric single-stranded RNA probes we were able to distinguish viral mRNA and viral double-stranded RNA components. We propose this technique as an ancillary or replacement procedure for the confirmation and classification of viral isolates using an appropriate battery of nucleic acid probes. Potential applications and methods of enhancing the technique are discussed.
Assuntos
Vírus Bluetongue/classificação , Hibridização de Ácido Nucleico , Reoviridae/classificação , Vírus Bluetongue/análise , Sondas RNA , RNA Viral/isolamento & purificação , Sorotipagem , SoluçõesRESUMO
Genome segments 2, 6, 8, and 9 of bluetongue virus (BTV) serotype 11, coding for P2, NS1, NS2, and P6, respectively, were cloned into pUC 8. Sizes of segment-2 and segment-6 clones indicated partial copies (55% and 80% of full length, respectively), whereas segment 8 and 9 clones represented full-length copies. Northern blot hybridizations of the clones to the 5 United States BTV prototypic serotypes (2, 10, 11, 13, and 17) revealed segment-2 clone to be serotype-specific to BTV-11, whereas segment 6, 8, and 9 clones were able to detect all serotypes to varying degrees. All clones failed to detect the related orbivirus, epizootic hemorrhagic disease virus.
Assuntos
Vírus Bluetongue/genética , RNA Viral/análise , Reoviridae/genética , Animais , Northern Blotting , Bluetongue/microbiologia , Clonagem Molecular , Sondas de DNA , Eletroforese em Gel de Ágar , Eletroforese em Gel de Poliacrilamida , Febres Hemorrágicas Virais/microbiologia , Febres Hemorrágicas Virais/veterinária , Hibridização de Ácido Nucleico , RNA de Cadeia Dupla/análise , RNA Viral/genética , Infecções por Reoviridae/microbiologia , Infecções por Reoviridae/veterinária , Células VeroRESUMO
A shotgun-cloning method incorporating all 10 bluetongue virus genome segments can simultaneously produce complete and partial copies of any of the genome segments. We report here 4 different cloned probes derived from 3 genome segments and individually defined by different hybridization recognition capabilities. One probe hybridized strongly with all 5 United States prototype strains of the 5 different bluetongue virus (BTV) serotypes existing in the United States and, as such, is a strong candidate for a broad BTV diagnostic probe in the United States. Another probe derived from genome segment 2 of BTV-17 hybridized only with the BTV-17 prototypic serotype, thereby demonstrating serospecific hybridization diagnostic potential. The implications for diagnostic and genetic relationship studies on BTV, using various genetic probes, are discussed.
Assuntos
Vírus Bluetongue/genética , Clonagem Molecular , Genes Virais , Reoviridae/genética , Animais , Linhagem Celular , Cricetinae , DNA/metabolismo , Rim , Hibridização de Ácido Nucleico , Plasmídeos , RNA de Cadeia Dupla/genética , RNA de Cadeia Dupla/isolamento & purificação , SorotipagemRESUMO
The gene regulatory circuitry through which pluripotent embryonic stem (ES) cells choose between self-renewal and differentiation appears vast and has yet to be distilled into an executive molecular program. We developed a data-constrained, computational approach to reduce complexity and to derive a set of functionally validated components and interaction combinations sufficient to explain observed ES cell behavior. This minimal set, the simplest version of which comprises only 16 interactions, 12 components, and three inputs, satisfies all prior specifications for self-renewal and furthermore predicts unknown and nonintuitive responses to compound genetic perturbations with an overall accuracy of 70%. We propose that propagation of ES cell identity is not determined by a vast interactome but rather can be explained by a relatively simple process of molecular computation.
Assuntos
Células-Tronco Embrionárias/metabolismo , Regulação da Expressão Gênica , Redes Reguladoras de Genes , Células-Tronco Pluripotentes/metabolismo , Fatores de Transcrição/metabolismo , Animais , Técnicas de Cultura de Células , Biologia Computacional , Camundongos , Fatores de Transcrição/genéticaRESUMO
Recombinant plasmids containing inserts representing genome segments 2, 5, 6, 8, and 9 of bluetongue virus (BTV) serotype 11, with tentative coding assignments for viral proteins P2, P5, NS1, NS2, and P6, respectively, have been used to study the genetic diversity within a BTV serotype using Northern blot hybridization. BTV 11 strains were isolated in California, Nevada, Oklahoma, and Mexico from elk, deer, and cattle. Diversity was indirectly indicated in the BTV 11 strains by comparisons of electropherotypes. Probes specific for segments 2, 6, 8, and 9 hybridized to all BTV 11 strains with only minor variation in hybridization signal. cDNA clones, representing 90 and 20% length copies of gene segment 5, detected a difference in the field isolates with hybridization signal correlating to mobility of this segment in SDS-PAGE. These two cDNA probes of genome segment 5 hybridized to BTV U.S. prototypic serotype 17 and not the remaining serotypes (BTV 2, 10, 13) or epizootic hemorrhagic disease virus (EHDV). These data suggest a close relationship between BTV-11 and 17 and/or alternatively are the result of genome segment reassortment between serotypes.
Assuntos
Vírus Bluetongue/genética , Variação Genética , RNA de Cadeia Dupla/análise , RNA Viral/análise , Reoviridae/genética , Animais , Autorradiografia , Northern Blotting , Vírus Bluetongue/classificação , California , Bovinos , Sondas de DNA , Cervos , Eletroforese em Gel de Poliacrilamida , México , Nevada , Hibridização de Ácido Nucleico , Oklahoma , Plasmídeos , Ruminantes , SorotipagemRESUMO
The nucleotide sequence of gene 5 encoding the rotavirus nonstructural protein NSP1 (NS53) of 6 strains (EW, EHP, RRV, I321, OSU, and Gottfried) was determined and compared to 6 previously reported strains (SA11, UK, RF, Hu803, DS-1, and Wa). The 12 rotavirus strains were derived from a total of five separate species (murine, bovine, simian, porcine, and human). Gene sizes ranged from 1564 to 1611 nucleotides in length and the deduced protein sequences were found to be 486 to 495 amino acids in length. Comparisons of NSP1 amino acid sequences showed identities ranging from 36 to 92%. This diversity was most evident between strains from different species. Phylogenetic analysis revealed a clustering of NSP1 sequences according to species origin with the exception that the human and porcine strains were included in a single grouping. Northern blot hybridizations using additional rotavirus strains from the five species confirmed the grouping found by sequence analysis. The species specificity of NSP1 is consistent with the hypothesis that NSP1 plays a role in host range restriction.
Assuntos
Proteínas de Ligação a RNA/química , Proteínas de Ligação a RNA/genética , Rotavirus/química , Rotavirus/genética , Proteínas não Estruturais Virais/química , Proteínas não Estruturais Virais/genética , Sequência de Aminoácidos , Sequência de Bases , Northern Blotting , Dados de Sequência Molecular , Análise de Sequência , Homologia de Sequência de Aminoácidos , Especificidade da EspécieRESUMO
The nucleotide sequence of genes 4 and 9, encoding the outer capsid proteins VP4 and VP7 of a serotype 10 tissue culture-adapted strain, I321, representative of asymptomatic neonatal rotaviruses isolated from neonates in Bangalore, India, were determined. Comparison of nucleotide and deduced amino acid sequences of I321 VP4 and VP7 with previously published sequences of various serotypes revealed that both genes were highly homologous to the respective genes of serotype 10 bovine rotavirus, B223. The VP4 of I321 represents a new human P serotype and the I321 and related strains represent the first description of neonatal rotaviruses that appear to derive both surface proteins from an animal rotavirus.
Assuntos
Antígenos Virais , Proteínas do Capsídeo , Capsídeo/genética , Rotavirus/classificação , Rotavirus/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Bovinos , Variação Genética , Humanos , Índia/epidemiologia , Recém-Nascido , Dados de Sequência Molecular , Rotavirus/crescimento & desenvolvimento , Homologia de Sequência de Aminoácidos , SorotipagemRESUMO
The antigenic structure of the VP4 protein of human rotavirus (HRV) strains Wa and ST3 was studied by using a panel of Wa- and ST3-derived VP4-specific neutralizing monoclonal antibodies (NMAbs) and NMAb-resistant variants. The VP4-coding genes from three Wa and three ST3 variants were sequenced. For Wa VP4, one homotypic and one heterotypic neutralization site, at amino acids 458 and 392, respectively, were identified. For ST3 VP4, three neutralization sites were found at amino acids 72, 217, and 385 that are either homotypic or associated with limited cross-reactivity. Cross-neutralization assays using several pairs of NMAbs and resistant variants showed that Wa VP4 has at least one large neutralization domain on its larger trypsin cleavage product, VP5*, consisting of several operationally related epitopes. VP4 of ST3 has at least two neutralization domains, one located on VP5* that is operationally related to the large neutralization domains on VP5* from HRVs Wa and KU, as well as an independent neutralization domain on VP8*, the smaller trypsin cleavage product of VP4.
Assuntos
Capsídeo/metabolismo , Rotavirus/metabolismo , Tripsina/metabolismo , Anticorpos Monoclonais/imunologia , Sequência de Bases , Sítios de Ligação de Anticorpos , Capsídeo/imunologia , Proteínas do Capsídeo , Reações Cruzadas , Primers do DNA , Epitopos/imunologia , Humanos , Hidrólise , Dados de Sequência Molecular , Testes de NeutralizaçãoRESUMO
Neutralizing monoclonal antibodies 2F1 and 1C10, which are specific for VP7 of serotype 2 rotaviruses (G2), were used to select neutralization escape variants of the human serotype 2 rotavirus, DS-1. Neutralization survival patterns generated by monoclonal antibodies 2F1, 1C10, and RV5:3 indicated that 2F1 and 1C10 did not recognize identical epitopes. Direct sequencing of PCR products of gene 8, encoding VP7, revealed that each escape variant possessed only a single nucleotide mutation which resulted in a single amino acid substitution. In one variant, a second nucleotide change occurred, but did not result in an amino acid change. Four independently selected 2F1 mutants showed mutations in four separate sites in antigenic regions A, C, and D. Two independently selected 1C10 variants had mutations in either the A region or an unreported site at amino acid 190. Two of the mutations resulted in the creation of new glycosylation sites which were utilized, but did not appear to greatly affect antigenicity. Of note, three of the mutants also demonstrated alterations in the migration patterns of gene 8 on PAGE. Such electrophoretic mobility shifts caused by single base neutralization escape mutations have not previously been reported for rotavirus. Since multiple mutations were selected with the same monoclonal antibody it appears that the antigenic regions of VP7, although widely separated in the linear sequence, are parts of a single, large and complex neutralization domain which includes amino acid 190 as well as the other previously reported epitopes.
Assuntos
Antígenos Virais/imunologia , Proteínas do Capsídeo , Capsídeo/imunologia , Epitopos/análise , Variação Genética , Mutação Puntual , Rotavirus/imunologia , Sequência de Aminoácidos , Anticorpos Monoclonais , Sequência de Bases , Capsídeo/biossíntese , Primers do DNA , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Genes Virais , Glicosilação , Humanos , Dados de Sequência Molecular , Testes de Neutralização , Reação em Cadeia da Polimerase , RNA Viral/análise , RNA Viral/metabolismo , Rotavirus/classificação , Rotavirus/genética , SorotipagemRESUMO
Bovine leukosis virus (BLV) is associated with the disease complex enzootic bovine leukosis. The infection may remain clinically silent in the form of an aleukaemic state or emerge as a persistent lymphocytosis and more rarely as lymphosarcroma. BLV has been considered classically to be a B lymphotropic virus, based upon the absolute increase in B lymphocytes in persistent lymphocytosis, the B lymphocyte phenotype of a majority of the cells making up lymphosarcomas and the identification of viral antigen expressed in B lymphocytes following in vitro culture of peripheral blood mononuclear leukocytes. This association of BLV with B lymphocytes is well established but the mechanism(s) of disease expression is not defined. To examine further the cellular tropism(s) of BLV, T lymphocyte subpopulations from 10 lymphocytotic cattle were established in vitro. Lymphocyte cultures were characterized by their subpopulation phenotype and DNA was extracted for identification of integrated provirus by Southern blot hybridization. Provirus was identified in T lymphocyte cultures derived from seven of 10 lymphocytotic cattle, with both T helper and T cytotoxic/suppressor subpopulations affected.
Assuntos
DNA Viral/análise , Vírus da Leucemia Bovina/genética , Leucemia/veterinária , Provírus/genética , Linfócitos T Citotóxicos/microbiologia , Linfócitos T Auxiliares-Indutores/microbiologia , Linfócitos T Reguladores/microbiologia , Animais , Southern Blotting , Bovinos , Doenças dos Bovinos/microbiologia , Células Cultivadas , Feminino , Citometria de Fluxo , Leucemia/microbiologia , Masculino , FenótipoRESUMO
The major neutralization antigen VP7 of rhesus rotavirus (RRV) was expressed in a baculovirus recombinant system. The expressed VP7 showed the same molecular mass as native VP7, and was recognized by hyperimmune sera as well as neutralizing and non-neutralizing monoclonal antibodies (MAbs) raised against RRV. Intraperitoneal administration of the expressed VP7 in mice elicited the production of serum antibodies which were able to immunoprecipitate VP7 from RRV-infected cell lysates and to neutralize the virus in vitro. Sera from immunized mice competed for binding to RRV in an ELISA with both neutralizing and non-neutralizing MAbs specific for VP7. Using a passive protection model of rotavirus disease, vaccination of mice with the recombinant VP7 induced partial protection from infection. These results suggest that the baculovirus-expressed VP7 may be useful in priming a protective immune response to rotavirus infection.
Assuntos
Anticorpos Antivirais/biossíntese , Antígenos Virais/imunologia , Proteínas do Capsídeo , Capsídeo/imunologia , Imunização , Infecções por Rotavirus/prevenção & controle , Rotavirus/imunologia , Animais , Animais Lactentes , Anticorpos Monoclonais , Especificidade de Anticorpos , Ligação Competitiva , Capsídeo/genética , Linhagem Celular , Feminino , Testes de Inibição da Hemaglutinação , Imunidade Materno-Adquirida , Macaca mulatta/virologia , Camundongos , Testes de Neutralização , Nucleopoliedrovírus/genética , Proteínas Recombinantes/imunologia , SpodopteraRESUMO
The nucleotide and deduced amino acid sequences of outer capsid proteins VP4 and VP7 of five murine rotavirus strains (EW, EB, EC, EL, EHP) were determined. Comparisons of the VP7 amino acid sequences of the five murine rotavirus strains with rotavirus strains representative of G serotypes 1-14 showed that the murine strains were highly homologous to one another and more closely related to strains representing G3 than to any other G type. Analysis of the VP4 amino acid sequences of the murine strains revealed the presence of at least two murine P types. Therefore, sequence analysis would predict that the murine rotaviruses are G3 or G3-like and represent at least two unique P types (tentatively P17 for EW, EB, EC, and EL and P18 for EHP). When we attempted to categorize the murine strains by serum neutralization tests, our results were less clear. Serum to two murine strains, EHP and EW, displayed one-way reactivity by focus-reduction neutralization assays using several prototype G3 strains. The G3 serotyping monoclonal antibody 159 failed to neutralize EHP and EW, while the G3-specific monoclonal YO-1E2 neutralized EHP, but not EW. A reassortant (A15) containing VP7 from EW and VP4 from RRV was neutralized by these two G3-specific monoclonal antibodies to a level 8- to 20-fold more than EW, but was still 64- to 250-fold less than either SA11 and RRV. These results suggest that VP4 can influence the antigenicity of VP7.
Assuntos
Antígenos Virais , Proteínas do Capsídeo , Capsídeo/química , Rotavirus/química , Sequência de Aminoácidos , Animais , Sequência de Bases , Capsídeo/imunologia , Bovinos , Cobaias , Humanos , Camundongos , Dados de Sequência Molecular , Testes de Neutralização , Rotavirus/imunologia , SorotipagemRESUMO
Rhesus rotavirus (RRV) VP4 trypsin cleavage product VP5(1)*, a truncated form of VP5*, was expressed in baculovirus and found by immunoprecipitation to be antigenically similar to VP5* on the virion. Immunization of mice with VP5(1)* elicited neutralizing antibody that was found to be cross-reactive with viruses representing P genotypes 1, 3, 4, 6, 7, and 8. Baculovirus expressed trypsin cleavage products, VP8* (amino acids 1-246) and VP5(1)* (amino acids 247-474), were tested for their ability to elicit a protective response in a murine model of passive protection. These results were compared to those obtained with baculovirus expressed RRV VP4. Dams immunized with baculovirus expressed RRV VP4 gave birth to pups protected from RRV virus challenge. Neither VP5(1)* nor VP8* was as effective at generating protective immunity as full length VP4. However, antibody to VP5(1)* was more effective than antibody to VP8* at mediating protection even though the neutralizing antibody titers as measured by hemagglutination inhibition and focus reduction neutralization were similar.
Assuntos
Antígenos Virais/genética , Proteínas do Capsídeo , Capsídeo/imunologia , Rotavirus/imunologia , Vacinas Sintéticas/imunologia , Animais , Antígenos Virais/química , Antígenos Virais/imunologia , Capsídeo/genética , Mapeamento de Epitopos , Feminino , Imunização Passiva , Camundongos , Nucleopoliedrovírus , Fragmentos de Peptídeos/imunologia , TripsinaRESUMO
Human rotaviruses were isolated from asymptomatic neonates at various hospitals and clinics in the city of Bangalore, India, and were found to be subgroup I specific and possess long RNA patterns (M. Sukumaran, K. Gowda, P. P. Maiya, T. P. Srinivas, M. S. Kumar, S. Aijaz, R. R. Reddy, L. Padilla, H. B. Greenberg, and C. D. Rao, Arch. Virol. 126:239-251, 1992). Three of these strains were adapted to tissue culture and found by serotype analysis and neutralization assays to be of serotype 10, a serotype commonly found in cattle but infrequently found in humans and not previously identified in neonates. By RNA-RNA hybridization, a high level of relatedness to a serotype 10 bovine rotavirus strain and a low-to-medium level of relatedness to a human rotavirus strain were observed. Since this human isolate shares a genogroup with bovine rotavirus, it is likely that it originated by interspecies transmission. A human rotavirus strain isolated from asymptomatic neonates and similar to bovine rotavirus might represent a good vaccine candidate.
Assuntos
Bovinos/microbiologia , Rotavirus/classificação , Rotavirus/genética , Animais , Anticorpos Monoclonais , Técnicas de Cultura , Genótipo , Humanos , Índia , Recém-Nascido , Hibridização de Ácido Nucleico , Sondas RNA , RNA Viral/análise , RNA Viral/genética , Sorotipagem , Especificidade da EspécieRESUMO
The development of general approaches for the isolation of efficient antivirals and the identification and validation of targets for drug screening are becoming increasingly important, due to the emergence of previously unrecognized viral diseases. The genetic suppressor element (GSE) technology is an approach based on the functional expression selection of efficient genetic inhibitors from random fragment libraries derived from a gene or genome of interest. We have applied this technology to isolate potent genetic inhibitors against HIV-1. Two strategies were used to select for GSEs that interfere with latent virus induction and productive HIV-1 infection based on the expression of intracellular and surface antigens. The selected GSEs clustered in seven narrowly defined regions of the HIV-1 genome and were found to be functionally active. These elements are potential candidates for the gene therapy of AIDS. The developed approaches can be applied to other viral pathogens, as well as for the identification of cellular genes supporting the HIV-1 life cycle.