RESUMO
Introduction: Fusarioid fungi that cause damping-off and root diseases can result in significant losses to conifer crops produced in forest nurseries across the USA. These nurseries are vital to reforestation and forest restoration efforts. Understanding the diversity of Fusarioid fungi associated with damping-off and root diseases of conifer seedlings can provide an approach for targeted management techniques to limit seedling losses and pathogen spread to novel landscapes. Methods: This study identifies 26 Fusarium spp. (F. acuminatum, F. annulatum, F. avenaceum, F. brachygibbosum, F. clavus, F. commune, F. cugenangense, F. diversisporum, F. elaeagni, F. elaeidis, F. flocciferum, F. fredkrugeri, F. fujikuroi, F. grosmichelii, F. ipomoeae, F. lactis, F. languescens, F. luffae, F. odoratissimum, F. oxysporum, F. queenslandicum, F. redolens, F. torulosum, F. triseptatum, F. vanleeuwenii, & F. verticillioides), 15 potential species within Fusarium and Neocosmospora species complexes (two from F. fujikuroi species complex, nine from F. oxysporum species complex, three from F. tricinctum species complex, and one from Neocosmospora species complex), and four Neocosmospora spp. (N. falciforme, N. metavorans, N. pisi, & N. solani) and associated host information collected from conifer-producing nurseries across the contiguous USA. Results: Phylogenetic analyses identified Fusarioid fungi haplotypes that were associated with 1) host specificity, 2) localization to geographic regions, or 3) generalists found on multiple hosts across diverse geographic regions. Discussion: The haplotypes and novel species identified on conifer seedlings should be considered for further analysis to determine pathogenicity, pathogen spread, and assess management practices.
RESUMO
Phosphonylmethoxyalkylpurine analogues were evaluated for their antitumor activity in murine tumor models. Three compounds, (S)-9-[(3-hydroxy-2-phosphonylmethoxy)propyl]adenine (HPMPA), 9-[(2-phosphonylmethoxy)ethyl]adenine (PMEA), and 9-[(2-phosphonylmethoxy)ethyl]guanine (PMEG) were modestly active with treated versus control (T/C) values of 125%-175% versus intraperitoneal P388 leukemia, but were inactive versus intravenously implanted P388. The most active and most potent of the three was PMEG, which was also evaluated against subcutaneously (SC) implanted B16 melanoma. In confirmatory experiments, optimal therapy with PMEG yielded reproducible increases in life span (T/C values of 164%-170%) and delays in primary tumor growth (7.3- to 13.0-day T-C values). PMEG is representative of a new class of antitumor antimetabolites heretofore recognized only for their antiviral properties.
Assuntos
Antineoplásicos , Guanina/análogos & derivados , Leucemia P388/tratamento farmacológico , Leucemia Experimental/tratamento farmacológico , Compostos Organofosforados/uso terapêutico , Animais , Relação Dose-Resposta a Droga , Guanina/administração & dosagem , Guanina/uso terapêutico , Injeções Intraperitoneais , Camundongos , Camundongos Endogâmicos , Transplante de Neoplasias , Compostos Organofosforados/administração & dosagem , Relação Estrutura-AtividadeRESUMO
An isoform of RGS9 was recently identified as the GTPase activating protein in bovine and mouse rod and cone photoreceptors. To explore the potential role of the RGS9 gene in human retinal disease, we determined its exon/intron arrangement, and investigated its expression in human retina. The results show that the gene, located on 17q24, consists of 19 exons and spans more than 75kb of genomic DNA. The entire gene was found to be contained on a single BAC clone with an insert size of 170kb. The major transcripts of the gene are alternatively spliced into a 9.5kb retina-specific transcript (RGS9-1) and a brain specific 2.5kb transcript (RGS9-2). Exons 1-16 are constitutive and present in both variants. Exon 17 contains the 3' end of the open reading frame and the 3'-UTR of the RGS9-1 variant. In RGS9-2, exon 17 is alternatively spliced and joined to exons 18 and 19 that are not present in the retina variant. Immunolocalization with a monoclonal antibody recognizing the retina and brain variants shows abundant expression in photoreceptors and possibly very low levels in cell types of the inner retina. Owing to the specific expression of RGS9-1 in photoreceptors the RGS9 gene is a candidate gene for RP17, a form of autosomal retinitis pigmentosa, located on the long arm of chromosome 17.
Assuntos
Processamento Alternativo , Genes/genética , Proteínas RGS/genética , Idoso , Sequência de Aminoácidos , Animais , Sequência de Bases , Southern Blotting , Western Blotting , Encéfalo/metabolismo , Pré-Escolar , Mapeamento Cromossômico , Cromossomos Humanos Par 17/genética , Corpo Estriado/química , Cricetinae , DNA/análise , DNA Complementar/genética , DNA Complementar/isolamento & purificação , Éxons , Regulação da Expressão Gênica no Desenvolvimento , Variação Genética , Humanos , Células Híbridas , Imuno-Histoquímica , Hibridização in Situ Fluorescente , Íntrons , Mamíferos/genética , Dados de Sequência Molecular , Isoformas de Proteínas/genética , Proteínas RGS/análise , Retina/química , Retina/metabolismo , Homologia de Sequência de AminoácidosRESUMO
The development of antibacterials was a very successful endeavor in the pharmaceutical company repertoire through the late 1970s, when interest in investing in antibiotic research and development temporarily waned. More recently, there have been a number of failures in late stage development or post-launch of human antibiotics. The answer to the dilemma of less-than-desired success may be the introduction of novel classes of agents, as well as development of new agents in traditional classes. This review provides an overview of the various "miscellaneous" antibacterials in development, excluding glycopeptides, macrolides, ketolides, and oxazolidinones. Among the agents highlighted in this review are the clinical candidates of quinolones, everninomycins, carbapenems, lipopeptides, glycylcyclines, and cephems. In several cases, certain quinolone agents described in this review will have been approved for marketing before press time.
Assuntos
Antibacterianos/farmacologia , Anti-Infecciosos/farmacologia , Avaliação Pré-Clínica de Medicamentos , Minociclina/análogos & derivados , 4-Quinolonas , Animais , Carbapenêmicos/farmacologia , Cefalosporinas/farmacologia , Ensaios Clínicos como Assunto , Daptomicina/farmacologia , Humanos , Testes de Sensibilidade Microbiana , Minociclina/farmacologia , Pró-Fármacos , Tetraciclinas/farmacologia , TigeciclinaRESUMO
The acyclic nucleotide analogue (S)-1-[3-hydroxy-2-(phosphonylmethoxy)propyl] cytosine (2, HPMPC) was prepared on a multigram scale in 18% overall yield starting from (R)-2,3-O-isopropylideneglycerol. The key step in the nine-step synthetic route is coupling of cytosine with the side-chain derivative 8 which bears a protected phosphonylmethyl ether group. In vitro data showed that HPMPC has good activity against herpes simplex virus types 1 and 2, although it was 10-fold less potent than acyclovir [AVC, 9-[(2-hydroxyethoxy)methyl]guanine]. By comparison, HPMPC exhibited greater activity than ACV against a thymidine kinase deficient strain of HSV 1 and was more potent than ganciclovir [DHPG, 9-[(1,3-dihydroxy-2-propoxy)methyl]guanine] against human cytomegalovirus. In vivo, HPMPC showed exceptional potency against HSV 1 systemic infection in mice, having an ED50 of 0.1 mg/kg per day (ip) compared with 50 mg/kg per day for ACV. HPMPC was also more efficacious than ACV in the topical treatment of HSV 1 induced cutaneous lesions in guinea pigs.
Assuntos
Antivirais/síntese química , Nucleotídeos de Citosina/síntese química , Citosina/análogos & derivados , Organofosfonatos , Compostos Organofosforados/síntese química , Animais , Fenômenos Químicos , Química , Cidofovir , Citomegalovirus/efeitos dos fármacos , Citosina/síntese química , Citosina/farmacologia , Nucleotídeos de Citosina/farmacologia , Feminino , Cobaias , Humanos , Camundongos , Compostos Organofosforados/farmacologia , Simplexvirus/efeitos dos fármacos , Células VeroRESUMO
A series of 2'-substituted derivatives of 9-[2-(phosphonomethoxy)ethyl]guanine (PMEG, 1) have been synthesized and evaluated in vitro for anti-human immunodeficiency virus (HIV) activity in the XTT assay and for anti-herpes activity in the plaque reduction assay. It has been observed that the anti-HIV activity of these derivatives depends on the size and the nature of the substituent as well as the chirality at the 2'-position of PMEG. In addition, these compounds generally demonstrated greater activity against HIV than herpes viruses. The most interesting analogues which emerged from these studies are (R)-2'-(azidomethyl)-PMEG [(R)-5] and (R)-2'-vinyl-PMEG [(R)-11]. The former showed anti-HIV activity with an IC50 of 5 microM and a cytotoxicity (CC50) greater than 1.4 mM in CEM cells. The latter has an IC50 of 13 microM for anti-HIV activity and a CC50 of greater than 1.6 mM. Furthermore, we have demonstrated that replacement of the guanine base of these 2'-substituted PMEG analogues with cytosine drastically reduces anti-HIV and anti-herpes activity.
Assuntos
Antivirais/síntese química , Guanina/análogos & derivados , Compostos Organofosforados/síntese química , Antineoplásicos/farmacologia , Antivirais/farmacologia , Guanina/farmacologia , Testes de Sensibilidade Microbiana , Compostos Organofosforados/farmacologia , Relação Estrutura-AtividadeRESUMO
A series of 9-(phosphonoalkyl)purines, which are analogues of 9-[2-(phosphonomethoxy)ethyl]purines (guanine, PMEG, 1; adenine, PMEA, 2), were synthesized. The analogues were tested for activity against herpes simplex virus types 1 and 2 (HSV-1 and HSV-2), human cytomegalovirus (HCMV), Rauscher murine leukemia virus (R-MuLV), and human immunodeficiency virus type 1 (HIV-1). With variations in the length of the alkyl chain, the optimal activity was achieved with two carbons between the purine base and the phosphonomethoxy functionality. Despite the structural similarity and the close pKa2 value of 8 to that of PMEG, no phosphorylation of 8 was observed by the bovine brain guanylate kinase. Since all isosteric analogues of PMEG (7-9) were not inhibitory against HSV-1 and HSV-2, the presence of the 3'-oxygen atom in the PME purines proved critical for anti-HSV activity. Introduction of the 1'-methyl group on the PMEG side chain significantly reduced its anti-HSV activity. Analogue 11, which is a mimic of the phosphate by incorporation of the alpha,alpha-difluoro carbon, was ineffective against HSV-1 and HSV-2. These results suggest that the structural requirements of PME purines for anti-HSV activity appear to be very strict.
Assuntos
Antivirais/síntese química , Organofosfonatos/síntese química , Purinas/síntese química , Fenômenos Químicos , Química , Cinética , Organofosfonatos/farmacologia , Purinas/farmacologia , Retroviridae/efeitos dos fármacos , Simplexvirus/efeitos dos fármacos , Relação Estrutura-AtividadeRESUMO
A number of methyl derivatives of 9-[2-(phosphonomethoxy)ethyl]guanine (PMEG, 1) have been synthesized and tested in vitro for anti-herpes and anti-human immunodeficiency virus (HIV) activity. Among these analogues, (R)-2'-methyl-PMEG [(R)-3] and 2',2'-dimethyl-PMEG (7) demonstrated potent anti-HIV activity in the XTT assay with EC50 values of 1.0 and 2.6 microM, respectively. The corresponding (S)-2'-methyl-PMEG [(S)-3] was found to be less potent against HIV. In addition, the (R) and (S) enantiomers of 9-[3-hydroxy-2-(phosphonomethoxy)propyl]guanine (HPMPG, 8) were prepared for comparison of biological activity, and shown to be active and equipotent against herpesviruses, but inactive against HIV.
Assuntos
Antivirais/farmacologia , Guanina/análogos & derivados , Compostos Organofosforados/farmacologia , Antivirais/síntese química , Antivirais/toxicidade , Linhagem Celular , Guanina/síntese química , Guanina/farmacologia , Guanina/toxicidade , HIV/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Compostos Organofosforados/síntese química , Compostos Organofosforados/toxicidade , Simplexvirus/efeitos dos fármacos , Estereoisomerismo , Relação Estrutura-AtividadeRESUMO
PURPOSE: To identify gene arrangement, chromosomal localization, and expression pattern of mouse guanylate cyclase activating proteins GCAP1 and GCAP2, retina-specific Ca2+-binding proteins, and photoreceptor guanylate cyclase activators. METHODS: The GCAP1 and GCAP2 genes were cloned from genomic libraries and sequenced. The chromosomal localization of the GCAP array was determined using fluorescent in situ hybridization. The expression of GCAP1 and GCAP2 in mouse retinal tissue was determined by immunocytochemistry. RESULTS: In this study, the mouse GCAP1 and GCAP2 gene array, its chromosomal localization, RNA transcripts, and immunolocalization of the gene products were fully characterized. The GCAP tail-to-tail array is located at the D band of chromosome 17. Each gene is transcribed into a single transcript of 0.8 kb (GCAP1) and 2 kb (GCAP2). Immunocytochemistry showed that both GCAP genes are expressed in retinal photoreceptor cells, but GCAP2 was nearly undetectable in cones. GCAP2 was also found in amacrine and ganglion cells of the inner retina. Light-adapted and dark-adapted retinas showed no significant difference in the distribution of the most intense GCAP2 staining within the outer segment and outer plexiform layers. CONCLUSIONS: Identical GCAP gene structures and the existence of the tail-to-tail gene array in mouse and human suggest an ancient gene duplication-inversion event preceding mammalian diversification. Identification of both GCAPs in synaptic regions, and of GCAP2 in the inner retina suggest roles of these Ca-binding proteins in addition to regulation of phototransduction.
Assuntos
Proteínas de Ligação ao Cálcio/genética , Proteínas de Ligação ao Cálcio/metabolismo , Células Fotorreceptoras/enzimologia , Adaptação Ocular , Sequência de Aminoácidos , Animais , Northern Blotting , Cromossomos/metabolismo , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Expressão Gênica , Proteínas Ativadoras de Guanilato Ciclase , Humanos , Hibridização in Situ Fluorescente , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Dados de Sequência Molecular , RNA Mensageiro/metabolismo , Retina/enzimologia , Células Ganglionares da Retina/enzimologia , Homologia de Sequência de AminoácidosRESUMO
PURPOSE: This study had three objectives: (1) to characterize the structures of the chicken GCAP1 and GCAP2 genes; (2) to determine if GCAP1, GCAP2, and GC1 genes are expressed in chicken pineal gland; (3) if GC1 is expressed in chicken pineal, to determine if the GC1 null mutation carried by the retinal degeneration (rd) chicken is associated with degenerative changes within the pineal glands of these animals. METHODS: GCAP1 and GCAP2 gene structures were determined by analyses of chicken cosmid and cDNA clones. The putative transcription start points for these genes were determined using 5'-RACE. GCAP1, GCAP2 and GC1 transcripts were analyzed using Northern blot and RT-PCR. Routine light microscopy was used to examine pineal morphology. RESULTS: Chicken GCAP1 and GCAP2 genes are arranged in a tail-to-tail array. Each protein is encoded by 4 exons that are interrupted by 3 introns of variable length, the positions of which are identical within each gene. The putative transcription start points for GCAP1 and GCAP2 are 314 and 243 bases upstream of the translation start codons of these genes, respectively. As in retina, GCAP1, GCAP2 and GC1 genes are expressed in the chicken pineal. Although the GC1 null mutation is present in both the retina and pineal of the rd chicken, only the retina appears to undergo degeneration. CONCLUSIONS: The identical arrangement of chicken, human, and mouse GCAP1/2 genes suggests that these genes originated from an ancient gene duplication/inversion event that occurred during evolution prior to vertebrate diversification. The expression of GC1, GCAP1, and GCAP2 in chicken pineal is consistent with the hypothesis that chicken pineal contains a functional phototransduction cascade. The absence of cellular degeneration in the rd pineal gland suggests that GC1 is not critical for pineal cell survival.
Assuntos
Proteínas de Ligação ao Cálcio/genética , Guanilato Ciclase/genética , Glândula Pineal/metabolismo , Receptores de Superfície Celular , Sequência de Aminoácidos , Animais , Animais Recém-Nascidos , Sequência de Bases , Northern Blotting , Proteínas de Ligação ao Cálcio/biossíntese , Galinhas/genética , Guanilato Ciclase/biossíntese , Proteínas Ativadoras de Guanilato Ciclase , Humanos , Camundongos , Dados de Sequência Molecular , Glândula Pineal/anatomia & histologia , Regiões Promotoras Genéticas , Degeneração Retiniana/genética , Degeneração Retiniana/patologia , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Our studies have shown that the acyclic nucleotide analogues PMEA and HPMPC are able to penetrate into cells and are then activated to mono- and diphosphate derivatives. The latter correspond to triphosphate analogues and presumably serve an important role in the biological activity exerted by these antiviral agents. In support of this idea, the inhibitory effect of PMEApp on HIV reverse transcriptase has been demonstrated with both RNA and DNA template-primer systems. Further studies will be undertaken to determine the effect of HPMPCpp on viral DNA polymerases. Whereas the metabolism of PMEA in CEM cells gives rise to only PMEAp and PMEApp, additional metabolites were obtained in MRC-5 cells; the identity of these metabolites remains to be determined. In the case of HPMPC, a third metabolite was obtained in addition to HPMPCp and HPMPCpp, which has been tentatively assigned as a phosphate-choline adduct by analogy with activation of cytosine-based nucleoside derivatives. The metabolism of HPMPC was unchanged between uninfected and infected cells, indicating that viral enzymes are not necessary for the activation of HPMPC. The long intracellular half-lives of the HPMPC metabolites may have implications for the antiviral efficacy of this compound. The persistence of activated metabolites suggests that infrequent dosing may be possible due to a prolonged antiviral effect. Our results on the effectiveness of infrequent dosing schedules with HPMPC in the treatment of HSV 2 infections in mice support this hypothesis. It is also possible that HPMPCp-choline may serve as a reservoir for HPMPC and therefore for the presumed active metabolite HPMPCpp.
Assuntos
Antivirais/farmacologia , HIV/efeitos dos fármacos , Nucleotídeos/farmacologia , Organofosfonatos , Adenina/análogos & derivados , Adenina/farmacologia , Células Cultivadas/efeitos dos fármacos , Cidofovir , Citosina/análogos & derivados , Citosina/farmacologia , Humanos , Compostos Organofosforados/farmacologiaRESUMO
General study design and environmental considerations. Scand j work environ health 9 (1983): suppl 1, 1-7. Recent concern regarding health hazards of methylene chloride stem primarily from the discovery of its metabolism to carboxyhemoglobin. In this report, a research program is described, the purpose of which was to assess potential health effects of methylene chloride exposure in an occupational setting. Particular attention was given to evaluating possible direct and carboxyhemoglobin-mediated effects on the hematopoietic and circulatory systems. The study involved one fiber production plant which used a methylene chloride/methanol mixture and acetone as solvents and a second fiber production plant that used acetone only. The research design included a retrospective cohort mortality study and several health evaluation studies, as well as an environmental assessment of the two plants. Industrial hygiene monitoring indicated that typical methylene chloride exposures ranged from an 8-h time-weighted average of 140 ppm in areas of low methylene chloride use to a corresponding average of 475 ppm in areas of high methylene chloride use and that methanol was present in about a one to ten ratio to methylene chloride. Acetone exposures in both plants ranged from 100 to over 1,000 ppm (time-weighted average).
Assuntos
Hidrocarbonetos Clorados/intoxicação , Cloreto de Metileno/intoxicação , Doenças Profissionais/induzido quimicamente , Adulto , Eletrocardiografia , Feminino , Inquéritos Epidemiológicos , Hematopoese/efeitos dos fármacos , Humanos , Masculino , Cloreto de Metileno/metabolismo , Pessoa de Meia-Idade , Doenças Profissionais/epidemiologia , Doenças Profissionais/metabolismo , Doenças Profissionais/mortalidade , Projetos de Pesquisa , Fumar , Solventes/intoxicaçãoRESUMO
Cedrus deodara is a highly valued conifer widely grown as an ornamental in the Pacific Northwest and southern United States. C. deodara in the Pacific Northwest is normally problem free but occasionally is damaged by dieback of shoot tips, which has been associated with a fungus resembling Sirococcus conigenus. In February 2002, bleeding cankers were observed on 2- to 4-year-old stems of potted nursery stock of C. deodara cv. Karl Fuchs from Clackamas County, OR. Cankers were dark with indistinct margins, shallow, and up to 30 cm long. Infection appeared to have originated with small twigs that had died. Cultures isolated from discolored bark on streptomycin-amended potato dextrose agar (PDA) produced conidiomata with hyaline, fusiform, two-celled conidia typical of S. conigenus (1,3). Inter-simple sequence repeat-polymerase chain reaction fingerprints of an isolate from one of these trees were consistent with the P group of S. conigenus (mostly from hosts in Picea and Pinus spp.) (2). This isolate (02-04, ATCC MYA-2969) was used to inoculate two shoots on each of 12 3-year-old potted deodar cedars in each of two trials. Removing a needle wounded each shoot, and an agar plug colonized with mycelium was placed over the wound and held in place for 2 weeks with Parafilm. Sterile agar plugs were applied to two wounded control shoots on each tree in each trial. After 10 weeks, 25 of 48 inoculated shoots were blighted and drooped with yellow to brown needles that eventually dropped. The pathogen was reisolated from 24 of 25 symptomatic shoots but not from asymptomatic or control shoots. To our knowledge, this is the first confirmed report of S. conigenus as a pathogen of C. deodara. References: (1) P. F. Cannon and D. W. Minter. Taxon 32:572, 1983. (2) D. R. Smith et al. For. Pathol. 33:141, 2003. (3) B. Sutton. The Coelomycetes. Commonw. Mycol. Inst., Kew, Surrey, England, 1980.
RESUMO
Despite the advent of newer imaging modalities, conventional radiography and clinical examination remain the primary screening method in evaluation of the mediastinum following blunt thoracic trauma. Mediastinal width (MW) is generally considered an important finding in assessing for aortic rupture. The degree of inspiration and patient positioning clearly affects MW, but is largely ignored in the literature. This study investigates the mediastinal widths of normal volunteers with differing degrees of inspiration and positioning, and compares them to radiographs of patients with known aortic ruptures. Mediastinal widths were obtained from chest radiographs of 16 patients with known aortic rupture, and from 50 volunteers using AP-inspiratory-supine, AP-expiratory-supine, and PA-inspiratory-upright technique. Upper 95% confidence limits were obtained for normals. A statistically significant difference in MW of normals was found between inspiratory-supine, expiratory-supine, and upright-inspiratory techniques. Compared to the same degree of inspiration in normals, 12 of 16 patients with aortic rupture had a MW above the upper 95% confidence limits. It is concluded that mediastinal width in normals is significantly affected by the degree of inspiration and positioning. When comparing mediastinal widths for normals and ruptures, there was a significant difference in MW for most degrees of inspiration. As depth of inspiration increased, differences between MW in controls and rupture patients increased. We conclude that patient positioning and degree of inspiration are important factors in assessing the mediastinum, and every effort should thus be made to obtain an upright-inspiratory film if clinically feasible prior to declaring a mediastinum as abnormal.
Assuntos
Ruptura Aórtica/diagnóstico por imagem , Mediastino/diagnóstico por imagem , Adolescente , Adulto , Estudos de Avaliação como Assunto , Feminino , Humanos , Masculino , Postura , Radiografia , Respiração , Supinação , Traumatismos Torácicos/diagnóstico por imagem , Ferimentos não Penetrantes/diagnóstico por imagemRESUMO
The rehabilitation of optimal masticatory function in many persons who have difficult prosthetic dental conditions can be significantly aided by newer orthognathic surgical procedures. The combined surgical-prosthetic approach for these patients requires close cooperation and communication between the general dentist or prosthodontist and the oral surgeon, particularly because various aspects of the surgical treatment planning for these patients differ greatly from those for patients with complete natural dentitions. A discussion and report of cases illustrate germane aspects of evaluation of conditions, treatment planning, and surgical approaches to the more commonly encountered surgical-prosthetic dental problems.
Assuntos
Dentaduras , Face/anormalidades , Anormalidades Maxilomandibulares/cirurgia , Adulto , Reabsorção Óssea/fisiopatologia , Cefalometria , Oclusão Dentária , Restauração Dentária Temporária , Estética Dentária , Face/cirurgia , Feminino , Humanos , Arcada Osseodentária/fisiologia , Anormalidades Maxilomandibulares/diagnóstico , Masculino , Mandíbula/cirurgia , Maxila/cirurgia , Pessoa de Meia-Idade , Osteotomia/métodos , Planejamento de Assistência ao Paciente , Periodonto/anatomia & histologia , Contenções , Extração DentáriaRESUMO
PURPOSE/AIM: To compare the efficacy of optical techniques with electrophysiological recordings for mapping retinal activity in response to electrical stimulation. MATERIALS AND METHODS: Whole cell patch clamp, Ca(2+) imaging (Fluo-4-AM), and Na(+) imaging (CoroNa Green-AM) techniques were used to detect responses of neurons from mouse and salamander retina to electrical stimulation. RESULTS: Synaptic currents were observed in ≥23% of retinal ganglion cells (RGCs), indicating presynaptic Ca(2+) increases in the inner plexiform layer (IPL). Modest depolarization with 20-30 mM K(+) consistently evoked Ca(2+) responses measured with Fluo4, but Ca(2+) responses were almost never evoked by epiretinal stimulation. In salamander retina, responses were seen in the inner nuclear layer (INL) and IPL. In mouse retina, responses were also sometimes seen in the outer pexiform layer (OPL). OPL responses showed a longer latency than IPL responses, suggesting that outer retinal circuits do not trigger synaptic responses of RGCs. Simultaneous Ca(2+) imaging and electrophysiological recording of synaptic currents confirmed that Fluo4-loaded retinas remained responsive to stimulation. Epiretinal stimulation evoked action potentials in ≥67% of RGCs. CoroNa Green detected Na(+) changes stimulated by 20 mM K(+), but epiretinal stimulation did not evoke detectable Na(+) responses. Simultaneous imaging and electrophysiological recording confirmed the health of CoroNa Green-loaded retinas. We confirmed stimulation efficacy by simultaneously recording Na(+) changes and electrophysiological responses. CONCLUSIONS: These data demonstrate that electrophysiological recordings show greater sensitivity than Na(+) or Ca(2+) imaging in response to electrical stimulation. The paucity of Ca(2+) responses is consistent with limited risk for Ca(2+)-mediated cell damage during electrical stimulation.