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1.
Fungal Biol ; 124(11): 958-968, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-33059847

RESUMO

Silicified fossil legume woods of Cynometroxylon Chowdhury & Ghosh collected from the Neogene (late Miocene) sediments of the Bengal Basin, eastern India, exhibit fungal decay seldom found in the fossil record. The wood possesses numerous perforate areas on the surface that seem to be the result of extensive fungal activity. In transverse section, the decayed areas (pockets) appear irregular to ellipsoidal in outline; in longitudinal section these areas of disrupted tissue are somewhat spindle-shaped. Individual pockets are randomly scattered throughout the secondary xylem or are restricted to a narrow zone. The aforesaid patterns of decay in fossil wood show similarities with that of white rot decay commonly produced by higher fungi, specifically basidiomycetes and ascomycetes. The host fossil wood harbors abundant ramifying and septate fungal hyphae with knob like swellings similar to pseudoclamps in basidiomycetes, and three-celled conidia-like reproductive structures. This record expands our current knowledge of wood decaying fungi-host plant interaction in the Neogene tropical forests of Peninsular India.


Assuntos
Basidiomycota , Fabaceae , Fósseis , Madeira , Fósseis/microbiologia , Índia , Madeira/microbiologia
2.
BJU Int ; 90(3): 216-23, 2002 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-12133055

RESUMO

OBJECTIVE: To assess the use of a thermo-expandable intraprostatic stent (Memokath(R), Engineers and Doctors A/S, Copenhagen, Denmark) for bladder outlet obstruction in men unable to undergo transurethral resection of the prostate (TURP), assessing symptoms, complications and duration of stent life. PATIENTS AND METHODS: The Memokath stent is a coil of a nickel-titanium alloy which has 'shape memory', the lower end expanding when heated to 55 degrees C. Risks associated with inserting the stent with a flexible cystoscope under local anaesthesia are minimal. Men were selected who were either permanently or temporarily unfit for TURP. Indications included severe respiratory and cardiovascular disease. Exclusion criteria included bladder carcinoma, calculi or detrusor failure; in all, 211 men were fitted with 217 intraprostatic stents over 8 years. RESULTS: There were 1511 TURPs during the study period; the mean age of men receiving a stent was 80.2 years, compared with 70.2 years for those undergoing TURP. The International Prostate Symptom Score decreased from a mean of 20.3 to 8.2 (P < 0.001) in the first 3 months after stent placement; there was virtually no change over 7 years. During the follow-up, 38% of men died with their stents in situ, 34% remain alive, 23% have had their stents removed for failure and 4% were removed as they were no longer required. There was a 13% migration rate and 16% repositioning rate. There were few side-effects (pain 3%, haematuria 3%, incontinence 6% and infection 6%). These frail men were more likely to die than have their stent fail. CONCLUSION: The Memokath intraprostatic stent is a valuable addition to the armamentarium of the urologist treating elderly or frail men with advanced bladder outlet obstruction and complements existing technologies.


Assuntos
Prostatectomia/instrumentação , Hiperplasia Prostática/cirurgia , Stents , Obstrução do Colo da Bexiga Urinária/cirurgia , Adulto , Idoso , Idoso de 80 Anos ou mais , Desenho de Equipamento , Humanos , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/etiologia , Prostatectomia/métodos , Hiperplasia Prostática/complicações , Análise de Sobrevida , Resultado do Tratamento , Obstrução do Colo da Bexiga Urinária/etiologia
3.
Int Immunol ; 13(9): 1109-20, 2001 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-11526091

RESUMO

This study examined whether therapy with a non-mitogenic, non-activating anti-CD3 mAb (G4.18) alone, or in combination with the T(h)2 cytokines, could inhibit induction or facilitate recovery from experimental allergic encephalomyelitis (EAE) in Lewis rats. G4.18, but not rIL-4, rIL-5 or anti-IL-4 mAb, reduced the severity and accelerated recovery from active EAE. A combination of rIL-4 with G4.18 was more effective than G4.18 alone. The infiltrate of CD4(+) and CD8(+) T cells, B cells, dendritic cells, and macrophages in the brain stem was less with combined G4.18 and IL-4 than G4.18 therapy or no treatment. Residual cells had preferential sparing of T(r)1 cytokines IL-5 and transforming growth factor-beta with loss of T(h)1 markers IL-2, IFN-gamma and IL-12Rbeta2, and the T(h)2 cytokine IL-4 as well as macrophage cytokines IL-10 and tumor necrosis factor-alpha. Lymph nodes draining the site of immunization had less mRNA for T(h)1 cytokines, but T(h)2 and T(r)1 cytokine expression was spared. Treatment with G4.18, rIL-4 or rIL-5 from the time of immunization had no effect on the course of active EAE. MRC OX-81, a mAb that blocks IL-4, delayed onset by 2 days, but had no effect on severity of active EAE. G4.18 also inhibited the ability of activated T cells from rats with active EAE to transfer passive EAE. This study demonstrated that T cell-mediated inflammation was rapidly reversed by a non-activating anti-CD3 mAb that blocked effector T(h)1 cells, and spared cells expressing T(h)2 and T(r)1 cytokines.


Assuntos
Anticorpos Monoclonais/uso terapêutico , Complexo CD3/imunologia , Encefalomielite Autoimune Experimental/terapia , Interleucina-4/uso terapêutico , Células Th1/efeitos dos fármacos , Animais , Tronco Encefálico/imunologia , Tronco Encefálico/patologia , Sinergismo Farmacológico , Imunização Passiva , Interleucina-4/imunologia , Interleucina-5/uso terapêutico , Leucócitos Mononucleares/patologia , Camundongos , Camundongos Endogâmicos BALB C , Proteína Básica da Mielina/imunologia , Ratos , Ratos Endogâmicos Lew , Proteínas Recombinantes/uso terapêutico , Medula Espinal/imunologia , Medula Espinal/patologia
4.
Antimicrob Agents Chemother ; 44(9): 2319-26, 2000 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-10952574

RESUMO

BMS-232632 is an azapeptide human immunodeficiency virus (HIV) type 1 (HIV-1) protease inhibitor that displays potent anti-HIV-1 activity (50% effective concentration [EC(50)], 2.6 to 5.3 nM; EC(90), 9 to 15 nM). In vitro passage of HIV-1 RF in the presence of inhibitors showed that BMS-232632 selected for resistant variants more slowly than nelfinavir or ritonavir did. Genotypic and phenotypic analysis of three different HIV strains resistant to BMS-232632 indicated that an N88S substitution in the viral protease appeared first during the selection process in two of the three strains. An I84V change appeared to be an important substitution in the third strain used. Mutations were also observed at the protease cleavage sites following drug selection. The evolution to resistance seemed distinct for each of the three strains used, suggesting multiple pathways to resistance and the importance of the viral genetic background. A cross-resistance study involving five other protease inhibitors indicated that BMS-232632-resistant virus remained sensitive to saquinavir, while it showed various levels (0. 1- to 71-fold decrease in sensitivity)-of cross-resistance to nelfinavir, indinavir, ritonavir, and amprenavir. In reciprocal experiments, the BMS-232632 susceptibility of HIV-1 variants selected in the presence of each of the other HIV-1 protease inhibitors showed that the nelfinavir-, saquinavir-, and amprenavir-resistant strains of HIV-1 remained sensitive to BMS-232632, while indinavir- and ritonavir-resistant viruses displayed six- to ninefold changes in BMS-232632 sensitivity. Taken together, our data suggest that BMS-232632 may be a valuable protease inhibitor for use in combination therapy.


Assuntos
Inibidores da Protease de HIV/farmacologia , Protease de HIV/metabolismo , HIV-1/efeitos dos fármacos , Oligopeptídeos/farmacologia , Piridinas/farmacologia , Sequência de Aminoácidos , Sulfato de Atazanavir , Resistência Microbiana a Medicamentos/fisiologia , Resistência a Múltiplos Medicamentos/fisiologia , Protease de HIV/genética , HIV-1/enzimologia , HIV-1/genética , Humanos , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , Mutação , Homologia de Sequência de Aminoácidos , Especificidade por Substrato
5.
J Assoc Nurses AIDS Care ; 9(3): 59-71, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-9589421

RESUMO

HIV/AIDS continues to pose a serious health hazard for gay men. Large numbers of men continue to become HIV infected each year, despite access to knowledge concerning how HIV is transmitted. Although gay men changed their sexual practices early in the epidemic, there is growing concern that there is a resurgence of risky sexual activities occurring in this group. Of particular concern is the resurgence of group sex activities. The purpose of this study was to explore the experiences of group sex in gay men to gain insight into the meaning of group sex to these men, the context in which it occurs, and men's views of how group sexual encounters relate to HIV transmission. The study used a qualitative approach to data collection. Ten self-identified gay men who reported engaging in group sex activities were interviewed concerning their experiences. Men reported that the most likely place for them to engage in group sex activities was in sex clubs, and a majority of their discussions centered on these clubs. Study data were analyzed using content analysis. Two overall categories of responses emerged from men's descriptions of their group sex experience: sexual desire and HIV/STD risk behaviors. Four themes--access to sex, sexual excitement or stimulation, sexual options, and control and sexual freedom--comprised the category of Sexual Desire. The themes identified within the category of HIV/STD risk behaviors included reframing risk, rejection of safer sex, and alcohol and drug use. The findings of the study suggest that sex in group settings such as sex clubs is a reality that must be addressed by HIV prevention efforts. Additionally, results indicate that current HIV prevention messages are being rejected by many gay men and need to be reevaluated for relevance two decades into the HIV/AIDS epidemic.


Assuntos
Infecções por HIV/prevenção & controle , Homossexualidade Masculina/psicologia , Assunção de Riscos , Parceiros Sexuais , Adulto , Atitude Frente a Saúde , Liberdade , Processos Grupais , Infecções por HIV/psicologia , Infecções por HIV/transmissão , Humanos , Relações Interpessoais , Entrevistas como Assunto , Masculino , Motivação , Comportamento Social , Sudeste dos Estados Unidos , Transtornos Relacionados ao Uso de Substâncias/psicologia , Saúde da População Urbana
6.
J Gen Virol ; 78 ( Pt 6): 1389-98, 1997 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-9191935

RESUMO

Degenerate and specific PCR assays were developed for bovine leukaemia virus (BLV) and/or primate T cell leukaemia/lymphoma viruses (PTLV). The degenerate assays detected all major variants of the BLV/PTLV genus at a sensitivity of 10-100 copies of input DNA; the specific systems detected 1-10 copies of input target. Sensitivity was 100% in specific DNA-PCR assays done on peripheral blood from seropositive BLV-infected cattle and HTLV-I- or HTLV-II-infected humans, and 62% in RNA/DNA-PCR assays on sera from BLV seropositive cattle. The pol fragments from 21 different BLV strains, isolated from cattle in North and Central America, were cloned and sequenced, and compared to other published BLV and PTLV pol sequences. BLV and PTLV sequences differed by 42%. Sequence divergence was up to 6% among the BLV strains, and up to 36% among the PTLV strains (with PTLV-I and PTLV-II differing among themselves by 15% and 8%, respectively). Some cows were infected with several BLV strains. Among retroviruses, BLV and PTLV sequences formed a distinct clade. The data support the interpretation that BLV and PTLV evolved from a common ancestor many millennia ago, and some considerable time before the PTLV-I and PTLV-II strains diverged from each other. The dissemination of the BLV strains studied probably resulted from the export of European cattle throughout the world over the last 500 years. The relatively similar mutation rates of BLV and PTLV, after their various points of divergence, suggest that there could be a much wider genetic range of BLV than has currently been defined.


Assuntos
Vírus da Leucemia Bovina/isolamento & purificação , Reação em Cadeia da Polimerase , Primatas/virologia , Retroviridae/isolamento & purificação , Sequência de Aminoácidos , Animais , Sequência de Bases , Bovinos , Linhagem Celular , Humanos , Vírus da Leucemia Bovina/classificação , Vírus da Leucemia Bovina/genética , Dados de Sequência Molecular , Filogenia , Retroviridae/classificação , Retroviridae/genética
8.
Transfusion ; 37(1): 52-9, 1997 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-9024490

RESUMO

BACKGROUND: Human T-cell lymphoma/leukemia viruses types I and II (HTLV-I and HTLV-II) are related exogenous human retroviruses. The former is definitely pathogenic while disease association with the latter is unclear. There are two subtypes of HTLV-II, A and B. Currently, enzyme-linked immunosorbent assays (ELISAs) based on HTLV-I antigens are used to screen for the presence of HTLV-I and -II antibodies. Confirmation and subtyping are accomplished by Western blot (WB) or ELISAs based on HTLV-I whole viral antigens and/or HTLV-I and HTLV-IIA peptides. The sensitivity and specificity of these serologic assays were compared to those of HTLV-I and-II-specific polymerase chain reaction (PCR) assays in tests on samples from Indians from South America in whom the HTLV-IIB subtype is endemic. STUDY DESIGN AND METHODS: Sera from 246 Gran Chaco Indians were evaluated for HTLV antibodies with the use of four ELISAs (Retrotek HTLV-I; Cambridge Biotech rgp21 enhanced HTLV-I/II; Vironostika HTLV-I/II; and Select HTLV-I/II), and a WB assay. Peripheral blood leukocyte DNA from each Indian was analyzed for HTLV-I or HTLV-II pol DNA via PCR. Fifteen of the PCR-positive samples were further subtyped via cloning and sequencing and/or oligomer restriction. RESULTS: Ninety-seven samples (39%) were positive for HTLV-II by serologic and/or PCR assays. All 15 positive DNA samples that were further analyzed were of the HTLV-IIB subtype and were clustered as a highly conserved phylogenetic group. Comparative analyses indicate that the sensitivity and specificity of the various assays were: PCR, 97 and 100 percent; Retrotek, 70 and 91 percent; Cambridge Biotech, 74 and 96 percent; Vironostika, 73 and 99 percent; Select 72 and 98 percent; and WB, 70 and 100 percent. CONCLUSION: The sensitivities of the tested HTLV serologic assays were comparable. However, the specificity of the Retrotek ELISA was significantly lower than that of the others. When positive, the subtyping assays were very specific. However, PCR assays would seem preferable or to be a necessary adjunct for the sensitive detection of HTLV-IIB infection.


Assuntos
Infecções por HTLV-II/etnologia , Infecções por HTLV-II/epidemiologia , Indígenas Sul-Americanos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Sequência de Aminoácidos , Western Blotting , Criança , Pré-Escolar , Ensaio de Imunoadsorção Enzimática , Reações Falso-Negativas , Soronegatividade para HIV , Soropositividade para HIV/diagnóstico , Infecções por HTLV-II/diagnóstico , Humanos , Pessoa de Meia-Idade , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Prevalência
9.
J Virol ; 69(6): 3675-82, 1995 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-7745716

RESUMO

Although a small amount of viral DNA has been shown to be enclosed within human immunodeficiency virus type 1 (HIV-1) virions, the majority of full-length viral DNA is formed after this virus infects target cells. Hence, we undertook investigations to identify the physical characteristics of the HIV-1 replication unit during the early events of infection. In these studies, nascent viral DNA synthesis was found to occur between 15 and 30 min after purified, DNase-treated HIV-1 virions were added to HUT 78 cells. At 1 h postinfection, a large amount of strong-stop viral DNA and some first-strand viral DNA had been synthesized. Several lines of evidence, including purification, nuclease digestion, and immunoprecipitation, indicated that these nascent viral DNAs were located within particles containing components such as reverse transcriptase and p24gag and gp120env proteins and having physical characteristics similar to those of intact virions.


Assuntos
HIV-1/genética , Transcrição Gênica , Vírion/genética , Linhagem Celular , DNA Viral/genética , HIV-1/fisiologia , HIV-1/ultraestrutura , Humanos , Cinética , Microscopia Eletrônica , Vírion/ultraestrutura , Replicação Viral
10.
J Med Virol ; 45(1): 1-9, 1995 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-7714483

RESUMO

There are 2 subtypes of human T-cell lymphoma/leukemia virus type II (HTLV-II), A and B. HTLV-II is increasingly associated with rare forms of lymphocytic neoplasia and a neurodegenerative disorder, characterized by hyperspasticity and ataxia. We have used PCR to amplify, clone and sequence 140 bp of the pol gene from many isolates of HTLV-IIA and HTLV-IIB from around the world. Analysis of these and other published sequence established that all HTLV-IIA sequences contained a unique Hinf I site and all HTLV-IIB sequences a unique Mse I site. A rapid and specific oligomer restriction (OR) assay was developed utilizing the primer pair SK110/SK111 and subsequent digestion with these enzymes. Concordance between sequenced and OR-based subtyping of DNA amplified by PCR was absolute among 22 HTLV-II isolates tested. Further OR or sequence analyses on an additional 30 other isolates indicated that the majority of North American non-indian HTLV-II isolates were subtype A, while all Paleo-Amerindian samples, including those from the Seminole of Florida; the Guaymi from Panama; and the Toba, Chorote, Wichi, and Chulupe of Argentina, belonged to subtype B. The SK110/SK111 PCR-OR format should facilitate molecular epidemiology studies of HTLV-II infection and allow for subtype stratification in assessing the sensitivity and specificity of HTLV detection formats and HTLV-II disease association.


Assuntos
DNA Viral/sangue , Infecções por HTLV-II/virologia , Vírus Linfotrópico T Tipo 2 Humano/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Polimorfismo de Fragmento de Restrição , Argentina , Sequência de Bases , Primers do DNA , Feminino , Genes pol/genética , Vírus Linfotrópico T Tipo 2 Humano/classificação , Humanos , Masculino , Dados de Sequência Molecular , Filogenia , Sensibilidade e Especificidade , Análise de Sequência de DNA , Homologia de Sequência do Ácido Nucleico , Especificidade da Espécie , Estados Unidos , Proteínas Virais/análise
11.
Aust N Z J Med ; 24(5): 554-60, 1994 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-7848159

RESUMO

BACKGROUND: Multiple organ transplants have become frequent. Combined heart-and-kidney grafting has been reported recently and we have pursued this in selected cases. AIMS: To devise a protocol for simultaneous heart-and-kidney transplantation, review our clinical experience with the procedure and the causes of cardiac and renal disease in this group. METHODS: Seven patients with advanced cardiac failure (LV ejection fraction < 0.29 units; five with IDCM), and chronic renal failure (serum creatinine > 375 mumol/L) due to a variety of causes, were accepted for combined heart-and-kidney transplantation. Four males, of mean age 33 years, underwent the procedure. Each received his organs from a single cadaveric donor with ABO blood group compatibility and a negative 'current' lymphocytotoxic cross-match, but without regard to HLA-antigen matching. Cardiac ischaemic time averaged 3 hours 40 minutes, the renal first warm time was 0 minutes in all cases, and renal cold and second warm ischaemic times averaged 5 hours 17 minutes and 52 minutes respectively. The heart was grafted first and the kidney second in a procedure which averaged seven hours. Immunosuppression was achieved by induction with antithymocyte globulin, thence steroids, azathioprine and cyclosporin A. RESULTS: No patient required post-operative dialysis. One patient had early urological complications requiring operative correction, but no serious opportunistic infections were observed. Early cardiac rejection on biopsy (ISHT grade 3a) was seen in three patients at four-ten weeks and responded promptly to increased steroids, but severe steroid-resistant rejection of both heart and kidney contemporaneously occurred in one of these three at 19 months and required a course of muromonab-CD3. All four patients developed hypertension. Mean creatinine clearance was 1.23 +/- 0.22 mL/second (74 +/- 13 mL/minute) at last follow-up. All four recipients were alive, well and rehabilitated 5, 20, 28 and 35 months after grafting. Two patients died while waiting for the double procedure and another patient eventually died after being taken off the dual waiting list and receiving a renal transplant only. CONCLUSIONS: In experienced hands, combined heart-and-kidney transplantation is feasible and offers a compelling therapeutic solution in the treatment of advanced cardiac and renal failure. IDCM is a frequent cause of the heart failure in this group.


Assuntos
Transplante de Coração , Transplante de Rim , Adolescente , Adulto , Feminino , Rejeição de Enxerto , Cardiopatias/complicações , Cardiopatias/cirurgia , Humanos , Falência Renal Crônica/complicações , Falência Renal Crônica/cirurgia , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias
13.
Am J Hematol ; 46(3): 234-40, 1994 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-7677806

RESUMO

Hemophilia B Leyden is a rare form of congenital factor IX deficiency which is characterized by severe factor IX deficiency at birth, which ameliorates after puberty. It is caused by mutations in the factor IX gene promoter region and the postpubertal amelioration is thought to be mediated by the action of testosterone on an androgen response element located in the promoter region. Three kindreds have been previously reported with a milder form of hemophilia B Leyden, associated with a guanine to adenine transition at nucleotide position -6 of the promoter region. We now report a fourth kindred with this mutation. The proband was a newborn with a factor IX level of 2.5%, his 12-year-old half-brother had a level of 28%, and his mother's 56-year-old maternal cousin had a level of 60%. A G to A transition at nucleotide -6 of the promoter region was demonstrated by cloning and sequencing polymerase chain reaction products from the half brother, and the mother was demonstrated to be a carrier. The mutation eliminates a TaqI restriction endonuclease site normally present in the wild type promoter, and the mother's cousin was demonstrated to carry the mutation by the absence of digestion with TaqI. The identification of hemophilia B Leyden with this specific mutation has practical importance to the clinical management because of its unique natural history and significantly better prognosis than classical hemophilia B.


Assuntos
Fator IX/genética , Sequência de Bases , Criança , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Mutação Puntual , Reação em Cadeia da Polimerase , Mapeamento por Restrição
14.
Virology ; 202(1): 379-89, 1994 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-7912021

RESUMO

In order to better understand the genomic diversity and molecular phylogeny of the human retroviruses, the plasmas from 250 Zairean patients collected in 1969 were tested for antibodies to human T-cell lymphoma and human immunodeficiency viruses (HTLV or HIV) using ELISA and confirmatory Western blots and for viral nucleic acids by reverse transcriptase-directed PCR (RT-PCR). Interestingly, none of the patients was confirmed positive for HIV, even though this region is now endemic for HIV-1. However, 74 (30%) and 3 (1%) of the samples were positive for antibodies to HTLV-I and II, respectively. Forty-four of 74 (59%) Western blot-positive Zairean samples were RT-PCR positive for HTLV-I, while 1 of 3 (33%) of HTLV-II-seropositive samples was RT-PCR positive. On the contrary, none of the Western blot-negative or indeterminate samples were RT-PCR positive for either HTLV-I or HTLV-II. We have cloned and sequenced 140 bp of the pol gene flanked by SK110/SK111 from 8 HTLV-I- and 1 HTLV-II-positive archival samples from Zaire. The HTLV-I isolates from Zaire cluster together as a phylogenetic group, diverging from the prototype Japanese HTLV-I (ATK) by a range of 1.4 to 3.6%. Their close homology to some African STLV-I isolates suggests relatively recent interspecies transmission. The Zairean HTLV-II isolate is closely grouped with the HTLV-II substrain of isolates found in Paleo-Amerindians of the New World, making it unlikely that it represents an endemic African strain.


Assuntos
Infecções por Deltaretrovirus/microbiologia , Deltaretrovirus/isolamento & purificação , Infecções por HIV/microbiologia , HIV/isolamento & purificação , Anticorpos Antivirais/sangue , Arquivos , Sequência de Bases , Western Blotting , DNA Viral/sangue , Deltaretrovirus/classificação , Deltaretrovirus/genética , Infecções por Deltaretrovirus/sangue , Infecções por Deltaretrovirus/epidemiologia , República Democrática do Congo/epidemiologia , Ensaio de Imunoadsorção Enzimática , Genes pol , HIV/classificação , HIV/genética , Infecções por HIV/sangue , Infecções por HIV/epidemiologia , Humanos , Vírus da Leucemia Bovina/genética , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , Prevalência
15.
J Virol Methods ; 46(1): 51-9, 1994 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-8175947

RESUMO

Signature primer pairs designed for use with the polymerase chain reaction have been developed which can determine if a positive result originated from the intended target nucleic acid or from so-called "carry-over" contamination of previously amplified DNA. The 3' ends of each signature primer, SK339/341, SSK110/111, and SSK58/59 contain a viral specific sequence complementary to regions of either HIV-1, HTLV-I and II respectively. The 5' ends of each primer contain a non-human, non-viral (NHNV) signature sequence including restriction endonuclease sites for subsequent cloning. A fourth set of primers, SK338/340, consist solely of these NHNV sequences and are designed to anneal to any product previously amplified by the viral-specific signature primers. These primers were tested against their corresponding positive and negative DNA targets, to determine their specificity and sensitivity. As expected, the viral-specific signature primers detected the retroviral infected samples while no detectable amplification occurred in negative DNA controls. Primers SK338/340 did not amplify any viral positive or negative template DNA's. Samples spiked with amplified material generated from the viral-specific signature primers could be specifically amplified by the NHNV primers SK338/340. Primers SK338/340 were determined to be more sensitive than the viral-specific signature primers, ensuring the detection of extremely low amounts of carryover. This strategy may be useful in developing other retroviral or non-retroviral primers with a built-in signature sequence that can differentiate false positives from true positives in a subsequent confirmatory test.


Assuntos
Artefatos , Primers do DNA , DNA Viral/isolamento & purificação , HIV-1/isolamento & purificação , Vírus Linfotrópico T Tipo 1 Humano/isolamento & purificação , Vírus Linfotrópico T Tipo 2 Humano/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Provírus/isolamento & purificação , Sequência de Bases , Linhagem Celular , Contaminação de Equipamentos , Reações Falso-Positivas , HIV-1/genética , Vírus Linfotrópico T Tipo 1 Humano/genética , Vírus Linfotrópico T Tipo 2 Humano/genética , Dados de Sequência Molecular , Provírus/genética , Sensibilidade e Especificidade
16.
AIDS Res Hum Retroviruses ; 9(12): 1287-96, 1993 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-8142146

RESUMO

Extracellular HIV-1 virions purified from cell culture supernatants have been found to contain viral DNA that is the result of partial reverse transcription within the virus particles. Our data supported these observations and further indicated that the ratio of genomic RNA to viral DNA was approximately 10(3):1 for the "strong stop" (R-U5) region and 10(5):1 for the gag region. We have shown that, in the absence of detergent, large amounts of DNase-resistant viral DNA can be synthesized within intact HIV-1 virions, indicating that this phenomenon is not dependent on perturbation of the viral envelope. Nascent viral DNA synthesis also occurred in purified virions incubated at 37 degrees C in cell-free human physiological fluids including seminal plasma, blood plasma, breast milk, and fecal fluid. In vitro HIV-1 infection assays, in which HIV-1 DNA synthesis was initiated in HIV-1 virions by prior incubation with deoxyribonucleoside triphosphates, demonstrated that virus particles so treated had an increased infectious titer over untreated virions when incubated with target human T cells. Our data suggest that HIV-1 virion-associated DNA synthesis may occur in vivo and may impact on the efficiency of intra- and interhost virus transmission. If so, this phenomenon should prove to be an important target for antiviral therapeutic strategies.


Assuntos
HIV-1/genética , Transcrição Gênica , Sequência de Bases , Líquidos Corporais/microbiologia , Primers do DNA/genética , DNA Viral/biossíntese , DNA Viral/genética , Desoxirribonucleotídeos/metabolismo , Fezes/microbiologia , Feminino , Genes gag , Infecções por HIV/microbiologia , Infecções por HIV/transmissão , HIV-1/metabolismo , Humanos , Técnicas In Vitro , Masculino , Leite Humano/microbiologia , Dados de Sequência Molecular , Plasma/microbiologia , Reação em Cadeia da Polimerase , RNA Viral/genética , Sêmen/microbiologia
17.
Cancer Res ; 53(24): 6067-73, 1993 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-8261424

RESUMO

Human T-cell lymphoma/leukemia virus type I (HTLV-I) causes adult T-cell leukemia/lymphoma and HTLV-I-associated myelopathy. Specific regions within the outer envelope proteins of other retroviruses, e.g., human immunodeficiency virus type 1, are highly immunogenic and, because of the selective pressure of the host immune system, quite variable. Mutations in the external envelope protein gene of murine retroviruses and human immunodeficiency virus type 1 influence cellular tropism and disease pathogenesis. By contrast, no disease-specific viral mutations have been identified in HTLV-I-infected patients. However, all isolates studied thus far have originated from leukemic cell lines, peripheral blood mononuclear cells, or cerebrospinal fluid lymphocytes from patients with HTLV-I-associated myelopathy and adult T-cell leukemia/lymphoma and, therefore, may not truly reflect tissue-associated variation. The midregion of the HTLV-I gp46 external envelope glycoprotein (amino acids 190-209) induces an antibody response in 90% of infected individuals, and a hexapeptide in this region (amino acids 191-196) elicits antibodies in rabbits which inhibit syncytia formation and infection of target lymphocytes. Because of the above, we expected the neutralizing domain of the gp46 env gene of HTLV-I to possess disease or organ-associated mutations selected by the infected host's immune system. Hence, we amplified, cloned, and sequenced HTLV-I DNA directly from in vivo central nervous system, spleen, and kidney specimens, and a leukemic cell line from a patient (M. J.) with both HTLV-I-associated myelopathy and adult T-cell leukemia/lymphoma to discern the possibility of tissue- and/or disease-specific variants. In addition, we sequenced several HTLV-I isolates from different regions of the world, including Papua New Guinea, Bellona, and Liberia, and compared them to other previously published HTLV-I and related retroviral sequences. The 239-base pair sequence corresponding to amino acids 178 to 256 in gp46 displayed minor tissue-specific variation in clones derived from central nervous system tissues from patient M. J., but overall was highly conserved at both the DNA and amino acid levels. Variation was observed in this region among the other HTLV-I, simian T-cell lymphoma virus type I, and HTLV-II isolates in a pattern that was consistent with their known phylogenetic relationship. No consistent disease-related changes were observed.(ABSTRACT TRUNCATED AT 400 WORDS)


Assuntos
Produtos do Gene env/química , Vírus Linfotrópico T Tipo 1 Humano/química , Leucemia de Células T/microbiologia , Proteínas de Neoplasias/química , Paraparesia Espástica Tropical/microbiologia , Proteínas Oncogênicas de Retroviridae/química , Vírus Linfotrópico T Tipo 1 de Símios/química , Sequência de Aminoácidos , Sequência de Bases , Produtos do Gene env/genética , Produtos do Gene env/imunologia , Vírus Linfotrópico T Tipo 1 Humano/isolamento & purificação , Humanos , Leucemia de Células T/complicações , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Paraparesia Espástica Tropical/complicações , Filogenia , Proteínas Oncogênicas de Retroviridae/genética , Proteínas Oncogênicas de Retroviridae/imunologia , Vírus Linfotrópico T Tipo 1 de Símios/isolamento & purificação
18.
J Virol ; 67(3): 1175-84, 1993 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-8437209

RESUMO

DNA from the peripheral blood mononuclear cells of 17 different individuals infected with human T-cell lymphoma/leukemia virus type II (HTLV-II) was successfully amplified by the polymerase chain reaction (PCR) with the primer pair SK110/SK111. This primer pair is conserved among the pol genes of all primate T-cell lymphoma viruses (PTLV) and flanks a 140-bp fragment of DNA which, when used in comparative analyses, reflects the relative degree of diversity among PTLV genomes. Cloning, sequencing, and phylogenetic comparisons of these amplified 140-bp pol fragments indicated that there are at least two distinct genetic substrains of HTLV-II in the Western Hemisphere. These data were confirmed for selected isolates by performing PCR, cloning, and sequencing with to 10 additional primer pair-probe sets specific for different regions throughout the PTLV genome. HTLV-II isolates from Seminole, Guaymi, and Tobas Indians belong in the new substrain of HTLV-II, while the prototype MoT isolate defines the original substrain. There was greater diversity among HTLV-II New World strains than among HTLV-I New World strains. In fact, the heterogeneity among HTLV-II strains from the Western Hemisphere was similar to that observed in HTLV-I and simian T-cell lymphoma/leukemia virus type I isolates from around the world, including Japan, Africa, and Papua New Guinea. Given these geographic and anthropological considerations and assuming similar mutation rates and selective forces among the PTLV, these data suggest either that HTLV-II has existed for a long time in the indigenous Amerindian population or that HTLV-II isolates introduced into the New World were more heterogeneous than the HTLV-I strains introduced into the New World.


Assuntos
Genes Virais/genética , Infecções por HTLV-II/genética , Vírus Linfotrópico T Tipo 2 Humano/genética , Polimorfismo Genético , Sequência de Aminoácidos , Argentina , Sequência de Bases , Genes env/genética , Genes pol/genética , Infecções por HTLV-I/epidemiologia , Infecções por HTLV-I/genética , Infecções por HTLV-II/epidemiologia , Vírus Linfotrópico T Tipo 1 Humano/classificação , Vírus Linfotrópico T Tipo 1 Humano/genética , Vírus Linfotrópico T Tipo 2 Humano/classificação , Humanos , Indígenas Sul-Americanos , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , Análise de Regressão , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido Nucleico , Estados Unidos
19.
Nurse Educ ; 17(4): 23-6, 1992.
Artigo em Inglês | MEDLINE | ID: mdl-1407733

RESUMO

The continued increase in reported cases of HIV disease has created a compelling need for nursing educators to provide students with comprehensive information on HIV-related issues. The authors discuss the need for students to care for HIV-positive individuals under faculty supervision to overcome fear and anxiety. Faculty education is proposed as a first step to develop a comprehensive approach to HIV education. A guide for integrating HIV content into the curriculum is provided.


Assuntos
Currículo , Educação em Enfermagem/normas , Infecções por HIV/enfermagem , HIV-1 , Competência Clínica/normas , Docentes de Enfermagem/normas , Humanos , Técnicas de Planejamento
20.
Burns Incl Therm Inj ; 13(1): 39-44, 1987 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-3548907

RESUMO

Despite improvements in functional rehabilitation secondary to better control of scar and contractures, aesthetic rehabilitation of the extensively burned face has remained a difficult problem. This study was undertaken to evaluate both technique and aesthetic results of early excision and split thickness autografting (STAG) of full skin thickness face burns. Twenty-five patients with full skin thickness face burns were operated on between days 4 and 14 post-burn. Thirteen patients had excision and STAG in one stage. Twelve patients had a two-stage procedure-excision and coverage with a biological dressing followed 24-72 h later by STAG. Seven of these patients had a pressure dressing in the form of a silicone face mask applied at the second stage. Early cosmetic results were encouraging in all patients. Twenty-five per cent of patients later required either contracture release or skin resurfacing. Preliminary results are encouraging and warrant evaluation by surgeons at other centres. When early excision of full skin thickness face burns is undertaken, cautious optimism as to the ultimate aesthetic result, both by the surgeon and the patient, is advisable.


Assuntos
Queimaduras/cirurgia , Traumatismos Faciais/cirurgia , Transplante de Pele , Cirurgia Plástica/métodos , Adolescente , Adulto , Criança , Pré-Escolar , Desbridamento , Estética , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Reoperação , Fatores de Tempo
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