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1.
Georgian Med News ; (322): 94-100, 2022 Jan.
Artigo em Russo | MEDLINE | ID: mdl-35134768

RESUMO

In a prospective randomized controlled longitudinal study, there have been assessed the results of early comprehensive rehabilitation of patients after total hip arthroplasty using "Levitas" passive suspension system. The study involved 85 patients (44 women and 41 men) with primary (77 people; 90,6%) and dysplastic coxarthrosis (8 people; 9,4%). The average age of the patients was 64,3±5,1 years old. Immediate results were evaluated in terms of pain intensity (visual-analogue scale), range of motion (M. D'Aubigne and M. Postel scales), functional activity (Harris scale) and mobility tests ("forward bend", "get up and go"), distant - by posturographic indicators and quality of life of the patients. All the patients were divided into 2 groups by simple randomization. The control group of the patients (42 people) had standard of rehabilitation measures (basic version of standard diet; prevention of early postoperative complications; drug support; alternating magnetic field; magnetic laser therapy; standard complex of physical therapy). The patients of the main group (43 people) additionally had treatment using a passive suspension system "Levitas" (Technomex, Poland). In both groups of patients, after a course of procedures, there has been recorded a significant decrease in pain, an increase in functional activity and an improvement in mobility tests (p <0.05-0.01). At the same time when comparing the final results obtained in the groups, the studied indicators had more significant shifts with the patients who received a complex treatment with additional prescription of "Levitas" sling system (p<0,05). According to the follow-up data, significant differences (p<0,05-0,01) were noted in stabilometric indicators and the quality of life (physical and mental health) which testified in favor of the patients in the main group. The absence of complications, side effects, deterioration in the condition of the patients in both groups indicates the safety of the used early rehabilitation methods. In the main group of the patients who additionally had exercises on the sling system, in comparison with the control group, there has been noted a more complete and high-quality restoration of the functional activity and support ability of the hip joint due to the early relief of pain syndrome, reducing in gait asymmetry, and optimization of the energy efficiency of body balance, which, apparently, is associated with adequate training of neuromuscular apparatus of both operated and contralateral limb, which provides more complete restructuring of the segments of the pelvic girdle.


Assuntos
Artroplastia de Quadril , Qualidade de Vida , Feminino , Humanos , Recém-Nascido , Estudos Longitudinais , Masculino , Plásticos , Estudos Prospectivos , Amplitude de Movimento Articular , Resultado do Tratamento
2.
Gynecol Oncol Rep ; 33: 100586, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32529019

RESUMO

•Serous borderline tumor outside of the peritoneal cavity is rare.•Involvement of cardiophrenic lymph nodes with serous borderline tumor can occur.•Preoperative imaging may aid surgical planning even in serous borderline tumor cases.•Sequencing can help confirm a diagnosis of serous borderline tumor at distant sites.

3.
Dalton Trans ; 46(48): 16783-16786, 2017 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-29165468

RESUMO

A unique phenomenon of regioisomerism in coordination chemistry was discovered: the reaction of a sterically hindered o-quinone annelated with a dithiete ring with Pd2dba3 in the presence of 1,2-bis(diphenylphosphino)ethane (dppe) gave a mixture of two regioisomers: catecholate 3Cat and dithiolate 3Dit. Both isomers were isolated in crystalline form and characterized by NMR, IR and X-ray diffractometry studies. DFT calculations reveal that the 3Dit species is more thermodynamically stable than the isomer 3Cat. Isomerization of 3Cat to 3Dit in solution was observed.

4.
Child Care Health Dev ; 43(6): 861-868, 2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-28744889

RESUMO

BACKGROUND: Cerebral palsy (CP) is the most common motor disability worldwide with an incidence of 2.5 per 1,000 births globally. Health beliefs among caregivers may be major drivers of health-related behaviours and service utilization, but little is known regarding health beliefs around CP in Africa. METHODS: Between July 2013 and September 2015, children with CP were identified in Gaborone, Botswana, and their caregivers were invited to participate in a qualitative study utilizing semistructured in-person one-on-one interviews. Interview questions addressed their understanding of CP, challenges of caring for a handicapped child, and community response to children with CP. RESULTS: Sixty-two caregivers participated in the study. Common themes elicited were variable knowledge about CP, financial and physical burden, lack of therapies and educational resources, and the impact of stigma. Caregivers in Botswana generally subscribed to a biomedical explanation of CP but expressed concerns regarding more stigmatizing folks beliefs expressed in the community. CONCLUSION: Health beliefs regarding CP in Botswana likely have a significant impact on utilization of healthcare resources. Information from this study should inform future educational interventions for caregivers of children with CP.


Assuntos
Cuidadores/psicologia , Paralisia Cerebral/psicologia , Conhecimentos, Atitudes e Prática em Saúde , Adolescente , Adulto , Botsuana , Paralisia Cerebral/etiologia , Criança , Pré-Escolar , Informação de Saúde ao Consumidor/organização & administração , Efeitos Psicossociais da Doença , Crianças com Deficiência/psicologia , Feminino , Folclore , Acessibilidade aos Serviços de Saúde/estatística & dados numéricos , Humanos , Entrevistas como Assunto , Masculino , Prognóstico , Pesquisa Qualitativa , Estigma Social , Adulto Jovem
5.
Dalton Trans ; 45(40): 15872-15878, 2016 Oct 12.
Artigo em Inglês | MEDLINE | ID: mdl-27301590

RESUMO

The reaction of (dpp-bian)AlEt(Et2O) (1) (dpp-bian = 1,2-bis[(2,6-diisopropylphenyl)imino]acenaphthene) with 4,4'-dimethoxybenzophenone leads to the replacement of Et2O with the ketone resulting complex (dpp-bian)AlEt[O[double bond, length as m-dash]C(4-MeOC6H4)2] (2). The reaction of compound 1 with 2,6-di-tert-butyl-4-methylphenol (1 : 1) or Ph2NH (1 : 1) affords the addition products [dpp-bian(H)]AlEt[O(2,6-tBu2-4-MeC6H2)] (3) and [dpp-bian(H)]AlEt(NPh2) (4) correspondingly. Treatment of complex 1 with 0.5 molar equivalents of H2O gives ethylalumoxane {[dpp-bian(H)]AlEt}2(µ-O) (5). Newly prepared complexes 2-5 have been characterized by NMR and IR spectroscopy, and the molecular structures of 2, 3, and 5 have been determined by single crystal X-ray analysis.

6.
Dalton Trans ; 44(47): 20532-41, 2015 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-26548805

RESUMO

In the presence of formic acid, acenaphthenequinone (AQ) reacts with one molar equivalent of 2,6-diisopropylaniline in toluene to give monoiminoacenaphtheneone (3, dpp-mian) in good yield. Reduction of compound 3 with an excess of magnesium in thf results in green crystalline amido-alcoholate [(dpp-mian)Mg(thf)2]2 (4). Crystallization of complex 4 from toluene affords a blue tetramer [(dpp-mian)Mg(thf)]4 (5). Reactions of compounds 4 and 5 with phenylacetylene proceed with C-C bond formation between the alkyne and the dpp-mian ligand to give the monomeric alkynyl-magnesium derivative [(dpp-mian)(PhC=CH2)Mg(C≡CPh)2(thf)]2 (7). Hydrolysis of complex 5 gives metal-free dpp-mian(PhC=CH2)H (8). Reaction of 7 with acetylacetone yields [{dpp-mian(PhC=CH2)}Mg(acac)]2 (9). Compounds 3-5 and 7-9 have been characterized by IR and NMR spectroscopy; molecular structures of 3, 5, 7, 8 and 9 have been determined by single crystal X-ray analysis.

7.
Vopr Onkol ; 59(2): 52-8, 2013.
Artigo em Russo | MEDLINE | ID: mdl-23814850

RESUMO

In this paper we present the results of a randomized prospective study, in which we examined the influence of rIL-2 ("Ronkoleykin ") on the effectiveness of the R-CHOP combination in the first-line treatment of patients with B-cell non-Hodgkin lymphomas. From March 2006 to December 2009, the study included 109 newly diagnosed patients. Standard R-CHOP was used in 59 patients, in 50 patients rIL-2 was added, injected subcutaneously in a dose of 1 million IU per day during each course in 1-5th days. It was found that the inclusion of rIL-2 in the R-CHOP regimen increased the effectiveness of the treatment of B-cell NHL with a high-risk of poor disease's course.


Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Interleucina-2/uso terapêutico , Linfoma de Células B/tratamento farmacológico , Adulto , Idoso , Anticorpos Monoclonais Murinos/administração & dosagem , Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Ciclofosfamida/administração & dosagem , Doxorrubicina/administração & dosagem , Esquema de Medicação , Sinergismo Farmacológico , Feminino , Humanos , Injeções Subcutâneas , Estimativa de Kaplan-Meier , Linfoma de Células B/patologia , Masculino , Pessoa de Meia-Idade , Prednisona/administração & dosagem , Estudos Prospectivos , Proteínas Recombinantes/uso terapêutico , Medição de Risco , Fatores de Risco , Rituximab , Resultado do Tratamento , Vincristina/administração & dosagem
8.
Angiol Sosud Khir ; 16(4): 93-6, 2010.
Artigo em Russo | MEDLINE | ID: mdl-21389951

RESUMO

The present study was aimed at assessing the patients' perception of the remote outcomes obtained after treatment for relapsing varicose disease. The patients treated for recurrent varicosity in 2007-2009 at the Minsk Municipal Centre for Vascular Surgery were encouraged to fill in the Chronic Venous Insufficiency Questionnaire (CIVIQ-2). The Questionnaire had been sent by post to a total of 67 operated patients, with the answers received from 40 (60%) subjects. The mean duration of the postoperative period amounted to 29.78 months. An excellent surgical outcome (i. е., no varicose veins present) was observed in seven responders, a satisfactory one was noted in sixteen patients (a minor, symptom-free relapse), an unsatisfactory outcome was noted in eleven subjects (a symptomatic relapse), and a poor one--in six. 57.9% of the patients considered their result as excellent to satisfactory and 42.1% turned out to be dissatisfied with the outcomes of the surgical intervention performed. The median of the CIVIQ-2 score in the group of the patients with satisfactory postoperative results amounted to 55.50 points [interquartile range (IQR) 27-87] as compared with 66.60 points (IQR 28-90) in the cohort of the patients dissatisfied with the therapeutic outcomes (P=0.009, Wilcoxon test). Hence, on average 29.78 months after surgery for a relapse of varicose disease, two thirds of the patients turned out to be satisfied with the outcomes of the treatment performed, with the remaining one third of the patients having remained dissatisfied therewith. Based on these findings, a conclusion was drawn that in all cases it is necessary to advise the patients presenting with varicose disease of possible long-term sequelae of the operation and possible development of recurrent varicosity.


Assuntos
Satisfação do Paciente , Qualidade de Vida , Varizes/psicologia , Adulto , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Recidiva , Estudos Retrospectivos , Inquéritos e Questionários , Varizes/cirurgia , Procedimentos Cirúrgicos Vasculares/psicologia
9.
J Exp Biol ; 210(Pt 15): 2607-17, 2007 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-17644675

RESUMO

Blood gases (P(O)2, P(CO)2, pH), oxygen content, hematocrit and hemoglobin concentration were measured during rest-associated apneas of nine juvenile northern elephant seals. In conjunction with blood volume determinations, these data were used to determine total blood oxygen stores, the rate and magnitude of blood O(2) depletion, the contribution of the blood O(2) store to apneic metabolic rate, and the degree of hypoxemia that occurs during these breath-holds. Mean body mass was 66+/-9.7 kg (+/- s.d.); blood volume was 196+/-20 ml kg(-1); and hemoglobin concentration was 23.5+/-1.5 g dl(-1). Rest apneas ranged in duration from 3.1 to 10.9 min. Arterial P(O)2 declined exponentially during apnea, ranging between a maximum of 108 mmHg and a minimum of 18 mmHg after a 9.1 min breath-hold. Venous P(O)2 values were indistinguishable from arterial values after the first minute of apnea; the lowest venous P(O)2 recorded was 15 mmHg after a 7.8 min apnea. O(2) contents were also similar between the arterial and venous systems, declining linearly at rates of 2.3 and 2.0 ml O(2) dl(-1) min(-1), respectively, from mean initial values of 27.2 and 26.0 ml O(2) dl(-1). These blood O(2) depletion rates are approximately twice the reported values during forced submersion and are consistent with maintenance of previously measured high cardiac outputs during rest-associated breath-holds. During a typical 7-min apnea, seals consumed, on average, 56% of the initial blood O(2) store of 52 ml O(2) kg(-1); this contributed 4.2 ml O(2) kg(-1) min(-1) to total body metabolic rate during the breath-hold. Extreme hypoxemic tolerance in these seals was demonstrated by arterial P(O)2 values during late apnea that were less than human thresholds for shallow-water blackout. Despite such low P(O)2s, there was no evidence of significant anaerobic metabolism, as changes in blood pH were minimal and attributable to increased P(CO)2. These findings and the previously reported lack of lactate accumulation during these breath-holds are consistent with the maintenance of aerobic metabolism even at low oxygen tensions during rest-associated apneas. Such hypoxemic tolerance is necessary in order to allow dissociation of O(2) from hemoglobin and provide effective utilization of the blood O(2) store.


Assuntos
Apneia/sangue , Oxigênio/sangue , Focas Verdadeiras/metabolismo , Animais , Apneia/metabolismo , Focas Verdadeiras/sangue
10.
Artigo em Inglês | MEDLINE | ID: mdl-16820312

RESUMO

In order to evaluate hemodynamics in the complex vascular system of phocid seals, intravascular pressure profiles were measured during periods of rest-associated apnea in young elephant seals (Mirounga angustirostris). There were no significant differences between apneic and eupneic mean arterial pressures. During apnea, venous pressure profiles (pulmonary artery, thoracic portion of the vena cava (thoracic vena cava), extradural vein, and hepatic sinus) demonstrated only minor, transient fluctuations. During eupnea, all venous pressure profiles were dominated by respiratory fluctuations. During inspiration, pressures in the thoracic vena cava and extradural vein decreased -9 to -21 mm Hg, and -9 to -17 mm Hg, respectively. In contrast, hepatic sinus pressure increased 2-6 mm Hg during inspiration. Nearly constant hepatic sinus and intrathoracic vascular pressure profiles during the breath-hold period are consistent with incomplete constriction of the caval sphincter during these rest-associated apneas. During eupnea, negative inspiratory intravascular pressures in the chest ("the respiratory pump") should augment venous return via both the venae cavae and the extradural vein. It is hypothesized that, in addition to the venae cavae, the prominent para-caval venous system of phocid seals (i.e., the extradural vein) is necessary to allow adequate venous return for maintenance of high cardiac outputs and blood pressure during eupnea.


Assuntos
Pressão Sanguínea/fisiologia , Coração/fisiologia , Focas Verdadeiras/fisiologia , Veias/fisiologia , Animais , Apneia/fisiopatologia , Artérias/fisiologia , Frequência Cardíaca/fisiologia , Modelos Cardiovasculares , Fluxo Sanguíneo Regional/fisiologia
11.
Artigo em Inglês | MEDLINE | ID: mdl-16574449

RESUMO

In order to evaluate hemodynamics and blood flow during rest-associated apnea in young elephant seals (Mirounga angustirostris), cardiac outputs (CO, thermodilution), heart rates (HR), and muscle blood flow (MBF, laser Doppler flowmetry) were measured. Mean apneic COs and HRs of three seals were 46% and 39% less than eupneic values, respectively (2.1+/-0.3 vs. 4.0+/-0.1 mL kg(-1) s(-1), and 54+/-6 vs. 89+/-14 beats min(-1)). The mean apneic stroke volume (SV) was not significantly different from the eupneic value (2.3+/-0.2 vs. 2.7+/-0.5 mL kg(-1)). Mean apneic MBF of three seals was 51% of the eupneic value. The decline in MBF during apnea was gradual, and variable in both rate and magnitude. In contrast to values previously documented in seals during forced submersions (FS), CO and SV during rest-associated apneas were maintained at levels characteristic of previously published values in similarly-sized terrestrial mammals at rest. Apneic COs of such magnitude and incomplete muscle ischemia during the apnea suggest that (1) most organs are not ischemic during rest-associated apneas, (2) the blood O(2) depletion rate is greater during rest-associated apneas than during FS, and (3) the blood O(2) store is not completely isolated from muscle during rest-associated apneas.


Assuntos
Apneia , Débito Cardíaco/fisiologia , Músculo Esquelético/irrigação sanguínea , Músculo Esquelético/fisiologia , Focas Verdadeiras/fisiologia , Animais , Frequência Cardíaca , Artéria Pulmonar/fisiopatologia , Fluxo Sanguíneo Regional/fisiologia , Especificidade da Espécie , Volume Sistólico , Temperatura , Fatores de Tempo
12.
Proc Natl Acad Sci U S A ; 98(17): 9814-8, 2001 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-11481427

RESUMO

We describe imaging the luminance of green fluorescent protein (GFP)-expressing bacteria from outside intact infected animals. This simple, nonintrusive technique can show in great detail the spatial-temporal behavior of the infectious process. The bacteria, expressing the GFP, are sufficiently bright as to be clearly visible from outside the infected animal and recorded with simple equipment. Introduced bacteria were observed in several mouse organs including the peritoneal cavity, stomach, small intestine, and colon. Instantaneous real-time images of the infectious process were acquired by using a color charge-coupled device video camera by simply illuminating mice at 490 nm. Most techniques for imaging the interior of intact animals may require the administration of exogenous substrates, anesthesia, or contrasting substances and require very long data collection times. In contrast, the whole-body fluorescence imaging described here is fast and requires no extraneous agents. The progress of Escherichia coli-GFP through the mouse gastrointestinal tract after gavage was followed in real-time by whole-body imaging. Bacteria, seen first in the stomach, migrated into the small intestine and subsequently into the colon, an observation confirmed by intravital direct imaging. An i.p. infection was established by i.p. injection of E. coli-GFP. The development of infection over 6 h and its regression after kanamycin treatment were visualized by whole-body imaging. This imaging technology affords a powerful approach to visualizing the infection process, determining the tissue specificity of infection, and the spatial migration of the infectious agents.


Assuntos
Antibacterianos/uso terapêutico , Colite/microbiologia , Sistema Digestório/microbiologia , Enterite/microbiologia , Infecções por Escherichia coli/microbiologia , Escherichia coli/isolamento & purificação , Corantes Fluorescentes , Fluorometria/métodos , Gastrite/microbiologia , Canamicina/uso terapêutico , Proteínas Luminescentes , Peritonite/microbiologia , Gravação de Videoteipe/métodos , Administração Oral , Animais , Antibacterianos/farmacologia , Colite/tratamento farmacológico , Colite/patologia , Sistemas Computacionais , Sistema Digestório/patologia , Progressão da Doença , Enterite/tratamento farmacológico , Enterite/patologia , Escherichia coli/efeitos dos fármacos , Infecções por Escherichia coli/tratamento farmacológico , Infecções por Escherichia coli/patologia , Feminino , Tecnologia de Fibra Óptica , Corantes Fluorescentes/análise , Fluorometria/instrumentação , Gastrite/tratamento farmacológico , Gastrite/patologia , Proteínas de Fluorescência Verde , Processamento de Imagem Assistida por Computador , Injeções Intraperitoneais , Canamicina/farmacologia , Iluminação , Proteínas Luminescentes/análise , Camundongos , Camundongos Nus , Peritonite/tratamento farmacológico , Peritonite/patologia , Gravação de Videoteipe/instrumentação
13.
J Opt Soc Am A Opt Image Sci Vis ; 18(5): 1186-91, 2001 May.
Artigo em Inglês | MEDLINE | ID: mdl-11336221

RESUMO

Waveguides in LiNbO3 are realized by a soft proton exchange (SPE) process with use of a melt of stearic acid highly diluted by lithium stearate. No phase transitions are formed when alpha-phase waveguides are obtained by SPE. The alpha-phase presents the same crystalline structure as that of pure LiNbO3 crystal, and it maintains the excellent nonlinear and electro-optical properties of the bulk material. The kinetics of the SPE method is studied by the use of secondary-ion mass spectrometry and prism-coupling techniques. The hydrogen effective diffusion coefficient as well as the self-diffusion coefficients of H+ and Li+ ions are determined as a function of the proton-exchange temperature for X-cut LiNbO3.

14.
Mol Ther ; 3(3): 319-22, 2001 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-11273773

RESUMO

Viral vectors based on lentiviruses, such as the human immunodeficiency virus, are able to transduce a broad spectrum of nondividing cells in vivo. This ability of lentiviral vectors makes them an attractive vehicle for gene transfer into the liver. In order to determine the requirements for efficient lentiviral gene transfer, we used a fluorescence imaging system, which allows the detection of cells and tissues that express fluorescent reporter genes (e.g., green fluorescence protein) in the living animal. We show that the latest generation of lentiviral vectors efficiently transduces the murine liver. Further analysis demonstrated that neither cell-cycle activation nor division of liver cells is a prerequisite for lentiviral gene transfer in vivo.


Assuntos
Técnicas de Transferência de Genes , Vetores Genéticos , Hepatócitos/metabolismo , Lentivirus/genética , Transdução Genética , Animais , Feminino , Genes Reporter/genética , Terapia Genética , Hepatócitos/citologia , Camundongos , Camundongos Nus , Transfecção , Integração Viral
15.
Proc Natl Acad Sci U S A ; 98(5): 2616-21, 2001 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-11226288

RESUMO

The development of drugs for the control of tumor angiogenesis requires a simple, accurate, and economical assay for tumor-induced vascularization. We have adapted the orthotopic implantation model to angiogenesis measurement by using human tumors labeled with Aequorea victoria green fluorescent protein for grafting into nude mice. The nonluminous induced capillaries are clearly visible against the very bright tumor fluorescence examined either intravitally or by whole-body luminance in real time. The orthotopic implantation model of human cancer has been well characterized, and fluorescence shadowing replaces the laborious histological techniques for determining blood vessel density. Intravital images of orthotopically implanted human pancreatic tumors clearly show angiogenic capillaries at both primary and metastatic sites. A quantitative time course of angiogenesis was determined for an orthotopically growing human prostate tumor periodically imaged intravitally in a single nude mouse over a 19-day period. Whole-body optical imaging of tumor angiogenesis was demonstrated by injecting fluorescent Lewis lung carcinoma cells into the s.c. site of the footpad of nude mice. The footpad is relatively transparent, with comparatively few resident blood vessels, allowing quantitative imaging of tumor angiogenesis in the intact animal. Capillary density increased linearly over a 10-day period as determined by whole-body imaging. Similarly, the green fluorescent protein-expressing human breast tumor MDA-MB-435 was orthotopically transplanted to the mouse fat pad, where whole-body optical imaging showed that blood vessel density increased linearly over a 20-week period. These powerful and clinically relevant angiogenesis mouse models can be used for real-time in vivo evaluation of agents inhibiting or promoting tumor angiogenesis in physiological microenvironments.


Assuntos
Neoplasias/irrigação sanguínea , Neovascularização Patológica , Animais , Feminino , Fluorescência , Proteínas de Fluorescência Verde , Humanos , Proteínas Luminescentes/genética , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Transplante de Neoplasias , Neoplasias/genética , Neoplasias/patologia , Células Tumorais Cultivadas
18.
Proc Natl Acad Sci U S A ; 97(22): 12278-82, 2000 Oct 24.
Artigo em Inglês | MEDLINE | ID: mdl-11050247

RESUMO

Transgene expression in intact animals now can be visualized by noninvasive techniques. However, the instruments and protocols developed so far have been formidable and expensive. We describe here a system for rapidly visualizing transgene expression in major organs of intact live mice that is simple, rapid, and eminently affordable. Green fluorescent protein (GFP) is expressed in the cells of brain, liver, pancreas, prostate, and bone, and its fluorescence is encoded in whole-body optical images. For low-magnification images, animals are illuminated atop a fluorescence light box and directly viewed with a thermoelectrically cooled color charge-coupled device camera. Higher-magnification images are made with the camera focused through an epi-fluorescence dissecting microscope. Both nude and normal mice were labeled by directly injecting 8 x 10(10) plaque-forming units/ml of adenoviral GFP in 20-100 microl PBS and 10% glycerol into either the brain, liver, pancreas, prostate, or bone marrow. Within 5-8 h after adenoviral GFP injection, the fluorescence of the expressed GFP in brain and liver became visible, and whole-body images were recorded at video rates. The GFP fluorescence continued to increase for at least 12 h and remained detectable in liver for up to 4 months. The system's rapidity of image acquisition makes it capable of real-time recording. It requires neither exogenous contrast agents, radioactive substrates, nor long processing times. The method requires only that the expressed gene or promoter be fused or operatively linked to GFP. A comparatively modest investment allows the study of the therapeutic and diagnostic potential of suitably tagged genes in relatively opaque organisms.


Assuntos
Perfilação da Expressão Gênica , Microscopia de Fluorescência/métodos , Adenoviridae/genética , Animais , Feminino , Genes Reporter , Proteínas de Fluorescência Verde , Proteínas Luminescentes/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Nus
19.
Pigment Cell Res ; 13(5): 345-51, 2000 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-11041211

RESUMO

In order to induce melanin production in mammalian cells with pigment disorders such as albino hair, a recombinant retrovirus containing the mel locus of Streptomyces antibioticus was constructed. The S. antibioticus mel locus, which consists of the open reading frame (ORF)-438 and the tyrosinase gene, was specifically derived by polymerase chain reaction (PCR) from Streptomyces plasmid pIJ702. The ORF-438 is required for the transfer of copper to apotyrosinase, which is essential for tyrosinase enzymatic activity. The tyrosinase gene was inserted into the XhoI/BamHI cloning site of the pLXSN retroviral vector to obtain pLtyrSN. An internal ribosome entry site (IRES) suitable for mammalian cell expression was obtained from the pLXIN retroviral vector by PCR. The ORF-438 and IRES DNA fragments were inserted into the pLtyrSN vector to obtain the tyrosinase-expression retroviral vector pLmelSN. The expression vector was amplified in murine PT67 packaging cells, where the ORF-438 and tyrosinase genes were also co-expressed as determined by reverse transcription-PCR. In order to evaluate the vector's ability to restore pigment production in cells with a pigment disorder, albino-mouse skins were histocultured and then infected with the pLmelSN retrovirus. Six days after infection, melanin granules were observed in approximately 60% of albino-mouse hair follicles in the histocultured skin. These results demonstrated that the S. antibioticus mel operon could express an active tyrosinase and produce melanin in the albino-mouse hair follicles. This novel gene therapy approach, using a small and simple tyrosinase operon in a high-expression vector, has a potentially wide application for therapy of pigment disorders in hair follicles.


Assuntos
Albinismo/terapia , Terapia Genética/métodos , Doenças do Cabelo/terapia , Monofenol Mono-Oxigenase/genética , Pele/metabolismo , Streptomyces antibioticus/enzimologia , Albinismo/genética , Animais , Técnicas de Cultura , Feminino , Expressão Gênica , Genes Bacterianos , Vetores Genéticos , Melaninas/biossíntese , Camundongos , Camundongos Endogâmicos C57BL , Monofenol Mono-Oxigenase/metabolismo , Fases de Leitura Aberta , RNA Mensageiro , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Retroviridae , Streptomyces antibioticus/genética
20.
Cancer Gene Ther ; 7(2): 332-8, 2000 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-10770644

RESUMO

We have previously demonstrated the antitumor efficacy of recombinant methioninase (rMETase) derived from Pseudomonas putida. To enhance the efficacy of rMETase, we have constructed the pLGFP-METSN retrovirus encoding the P. putida methioninase (MET) gene fused with the green fluorescent protein (GFP) gene. pLGFP-METSN or control vector pLGFPSN was introduced into the human lung cancer cell line H460. The methionine level of H460-GFP-MET cells was reduced to 33% of that of H460-GFP cells. rMETase (0.08 U/mL) in the medium resulted in 10% survival of H460-GFP-MET cells compared with untreated cells in vitro. In contrast, rMETase-treated H460-GFP cells survived at 90% of control. Tissue fragments harvested from subcutaneous tumors of H460-GFP-MET or H460-MET were implanted by surgical orthotopic implantation into the lungs of nude mice. A suboptimal dose of rMETase was administered intraperitoneally daily to mice in each group. Overall survival of rMETase-treated animals with H460-GFP-MET tumors was significantly longer than either rMETase-treated or -untreated animals with H460-GFP tumors (P < .05 in log-rank test). In two repeat experiments, rMETase-treated animals with H460-GFP-MET tumors had a 30-day survival of 80% and 83%, respectively. Untreated animals with H460-GFP-MET tumors had a 30-day survival of 40% and 58%, respectively. rMETase-treated animals with H460-GFP tumors had a 30-day survival of 0% and 33%, respectively. Untreated animals with H460-GFP tumors had a 30-day survival of 0% and 33%, respectively. The retrovirus-mediated gene transfer of METase decreased the intracellular methionine level of tumor cells and consequently enhanced the efficacy of treatment with the rMETase protein. We have thus demonstrated a new strategy of combination tumor therapy with the gene and gene product of MET.


Assuntos
Antimetabólitos Antineoplásicos/uso terapêutico , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Liases de Carbono-Enxofre/genética , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/terapia , Animais , Liases de Carbono-Enxofre/metabolismo , Feminino , Vetores Genéticos/genética , Proteínas de Fluorescência Verde , Humanos , Proteínas Luminescentes/genética , Neoplasias Pulmonares/enzimologia , Neoplasias Pulmonares/mortalidade , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Transplante de Neoplasias , Retroviridae/genética , Taxa de Sobrevida , Transfecção , Células Tumorais Cultivadas
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