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1.
Opt Express ; 31(26): 43035-43047, 2023 Dec 18.
Artigo em Inglês | MEDLINE | ID: mdl-38178406

RESUMO

The scalability of quantum networking will benefit from quantum and classical communications coexisting in shared fibers, the main challenge being spontaneous Raman scattering noise. We investigate the coexistence of multi-channel O-band quantum and C-band classical communications. We characterize multiple narrowband entangled photon pair channels across 1282 nm-1318 nm co-propagating over 48 km of installed standard fiber with record C-band power (>18 dBm) and demonstrate that some quantum-classical wavelength combinations significantly outperform others. We analyze the Raman noise spectrum, optimal wavelength engineering, multi-photon pair emission in entangled photon-classical coexistence, and evaluate the implications for future quantum applications.

2.
Opt Lett ; 46(11): 2730-2733, 2021 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-34061099

RESUMO

We demonstrate dual-channel phase-shifted Bragg grating filters in the telecom band on thin-film lithium niobate. These integrated tunable ultra-narrow linewidth filters are crucial components for optical communication and sensing systems, as well as future quantum-photonic applications. Thin-film lithium niobate is an emerging platform suitable for these applications and has been exploited in this Letter. The demonstrated device has an extinction ratio of 27 dB and two channels with close linewidths of about 19 pm (quality factor of ${8} \times {{10}^4}$), separated by 19 GHz. The central wavelength could be efficiently tuned using the high electro-optic effect in lithium niobate with a tuning factor of 3.83 pm/V. This demonstration can be extended to tunable filters with multiple channels, along with desired frequency separations and optimized tunability, which would be useful for a variety of complex photonic integrated circuits.

3.
Am J Obstet Gynecol ; 225(5): 475.e1-475.e19, 2021 11.
Artigo em Inglês | MEDLINE | ID: mdl-34087227

RESUMO

OBJECTIVE: Women consider preservation of sexual activity and improvement of sexual function as important goals after pelvic organ prolapse surgery. This systematic review aimed to compare sexual activity and function before and after prolapse surgery among specific approaches to pelvic organ prolapse surgery including native tissue repairs, transvaginal synthetic mesh, biologic grafts, and sacrocolpopexy. DATA SOURCES: MEDLINE, Embase, and ClinicalTrials.gov databases were searched from inception to March 2021. STUDY ELIGIBILITY CRITERIA: Prospective comparative cohort and randomized studies of pelvic organ prolapse surgeries were included that reported the following specific sexual function outcomes: baseline and postoperative sexual activity, dyspareunia, and validated sexual function questionnaire scores. Notably, the following 4 comparisons were made: transvaginal synthetic mesh vs native tissue repairs, sacrocolpopexy vs native tissue repairs, transvaginal synthetic mesh vs sacrocolpopexy, and biologic graft vs native tissue repairs. METHODS: Studies were double screened for inclusion and extracted for population characteristics, sexual function outcomes, and methodological quality. Evidence profiles were generated for each surgery comparison by grading quality of evidence for each outcome across studies using a modified Grading of Recommendations Assessment, Development and Evaluation (GRADE) system. RESULTS: Screening of 3651 abstracts was performed and identified 77 original studies. The overall quality of evidence was moderate to high. There were 26 studies comparing transvaginal synthetic mesh with native tissue repairs, 5 comparing sacrocolpopexy with native tissue repairs, 5 comparing transvaginal synthetic mesh with sacrocolpopexy, and 7 comparing biologic graft with native tissue repairs. For transvaginal synthetic mesh vs native tissue repairs, no statistical differences were found in baseline or postoperative sexual activity, baseline or postoperative total dyspareunia, persistent dyspareunia, and de novo dyspareunia. Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire short form change scores were not different between transvaginal synthetic mesh and native tissue repairs (net difference, -0.3; 95% confidence interval, -1.4 to 0.8). For sacrocolpopexy vs native tissue repairs, baseline or postoperative sexual activity, baseline or postoperative total dyspareunia, de novo dyspareunia, and Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire short form score differences were not different. For biologic graft vs native tissue repairs, baseline or postoperative sexual activity, baseline or postoperative total dyspareunia, and Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire short form changes were also not different. For transvaginal synthetic mesh vs sacrocolpopexy, there was no difference in sexual activity and sexual function score change. Based on 2 studies, postoperative total dyspareunia was more common in transvaginal synthetic mesh than sacrocolpopexy (27.5% vs 12.2%; odds ratio, 2.72; 95% confidence interval, 1.33-5.58). The prevalence of postoperative dyspareunia was lower than preoperative dyspareunia after all surgery types. CONCLUSION: Sexual function comparisons are most robust between transvaginal synthetic mesh and native tissue repairs and show similar prevalence of sexual activity, de novo dyspareunia, and sexual function scores. Total dyspareunia is higher after transvaginal synthetic mesh than sacrocolpopexy. Although sexual function data are sparse in the other comparisons, no other differences in sexual activity, dyspareunia, and sexual function score change were found.


Assuntos
Dispareunia/etiologia , Procedimentos Cirúrgicos em Ginecologia , Prolapso de Órgão Pélvico/cirurgia , Feminino , Procedimentos Cirúrgicos em Ginecologia/efeitos adversos , Humanos , Complicações Pós-Operatórias , Telas Cirúrgicas
4.
Opt Express ; 28(26): 39963-39975, 2020 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-33379534

RESUMO

We generate quantum-correlated photon pairs using cascaded χ(2):χ(2) traveling-wave interactions for second-harmonic generation (SHG) and spontaneous parametric down-conversion (SPDC) in a single periodically-poled thin-film lithium-niobate (TFLN) waveguide. When pulse-pumped at 50 MHz, a 4-mm-long poled region with nearly 300%/Wcm2 SHG peak efficiency yields a generated photon-pair probability of 7±0.2 × 10-4 with corresponding coincidence-to-accidental ratio (CAR) of 13.6±0.7. The CAR is found to be limited by Stokes/anti-Stokes Raman-scattering noise generated primarily in the waveguide. A Raman peak of photon counts at 250 cm-1 Stokes shift from the fundamental-pump wavenumber suggests most of the noise that limits the CAR originates within the lithium niobate material of the waveguide.

5.
Opt Express ; 27(26): 38521-38531, 2019 Dec 23.
Artigo em Inglês | MEDLINE | ID: mdl-31878617

RESUMO

An efficient source of quantum-correlated photon-pairs that is integrable with existing silicon-electronics fabrication techniques is desirable for use in quantum photonic integrated circuits. Here we demonstrate signal-idler photon pairs with high coincidence-to-accidental count ratios of over 103 on a coarse wavelength-division-multiplexing grid that spans 140 nm by using a 300-µm-long poled region in a thin-film periodically-poled lithium-niobate ridge waveguide bonded to silicon. The pairs are generated via spontaneous parametric downconversion pumped by a continuous-wave tunable laser source. The small mode area of the waveguide allows for efficient interaction in a short length of the waveguide and, as a result, permits photon-pair generation over a broad range of signal-idler wavelengths.

6.
Opt Lett ; 42(5): 951-954, 2017 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-28248339

RESUMO

We experimentally demonstrate the temporal reshaping of optical waveforms in the telecom wavelength band using the principle of quantum frequency conversion. The reshaped optical pulses do not undergo any wavelength translation. The interaction takes place in a nonlinear χ(2) waveguide using an appropriately designed pump pulse programmed via an optical waveform generator. We show the reshaping of a single-peak pulse into a double-peak pulse and vice versa. We also show that exponentially decaying pulses can be reshaped into a near Gaussian shape, and vice versa, which is a useful functionality for quantum communications.

7.
Int Urogynecol J ; 28(2): 249-256, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27581769

RESUMO

INTRODUCTION: Interstitial cystitis/bladder pain syndrome (IC/BPS) is a poorly understood source of chronic pain causing significant morbidity, with variable treatment success. Despite the need to understand patient perspectives in chronic pain, there is a paucity of qualitative data for IC/BPS. We aimed to acquire information regarding patient experience with IC/BPS symptoms and with their medical care to elicit suggestions to improve patient satisfaction with that care. METHODS: Fifteen women with IC/PBS participated in a total of four focus groups. Sessions were recorded and transcribed and information deidentified. Focus groups were conducted until thematic saturation was reached. All transcripts were coded and analyzed by a minimum of three independent physician reviewers. Investigators identified emergent themes and concepts using grounded-theory methodology. RESULTS: Participant's mean age was 52.6 years, with an average IC/BPS duration of 6.3 years. Thematic saturation was reached after four focus groups. We identified three emergent patient experience concepts: IC/PBS is debilitating, the disease course is unpredictable and unrelenting, and patients experience significant isolation. Importantly, suicidal ideation was expressed in each group. Patients voiced strong preference for physicians who provided education regarding the condition, an array of treatment options, organized treatment plans, and optimism and hope regarding treatment outcomes. CONCLUSIONS: Our study presents novel findings of the importance of patient-physician interaction in IC/BPS and reinforces the tremendous disability and burden of this disease, which frequently manifests in suicidal ideation. Patients preferred organized treatment plans with diverse choices and providers who offered hope in dealing with their condition.


Assuntos
Dor Crônica/psicologia , Cistite Intersticial/psicologia , Satisfação do Paciente , Relações Médico-Paciente , Qualidade de Vida , Adulto , Ansiedade/psicologia , Depressão/psicologia , Feminino , Grupos Focais , Humanos , Pessoa de Meia-Idade , Assistência Centrada no Paciente , Pesquisa Qualitativa , Isolamento Social/psicologia , Ideação Suicida , Síndrome
8.
Am J Obstet Gynecol ; 215(5): 654.e1-654.e10, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27319368

RESUMO

BACKGROUND: Internet resources are becoming increasingly important for patients seeking medical knowledge. It is imperative to understand patient use and preferences for using the Internet and social networking websites to optimize patient education. OBJECTIVES: The purpose of this study was to evaluate social networking and Internet use among women with pelvic floor complaints to seek information for their conditions as well as describe the likelihood, preferences, and predictors of website usage. STUDY DESIGN: This was a cross-sectional, multicenter study of women presenting to clinical practices of 10 female pelvic medicine and reconstructive surgery fellowship programs across the United States, affiliated with the Fellows' Pelvic Research Network. New female patients presenting with pelvic floor complaints, including urinary incontinence, pelvic organ prolapse, and fecal incontinence were eligible. Participants completed a 24 item questionnaire designed by the authors to assess demographic information, general Internet use, preferences regarding social networking websites, referral patterns, and resources utilized to learn about their pelvic floor complaints. Internet use was quantified as high (≥4 times/wk), moderate (2-3 times/wk), or minimal (≤1 time/wk). Means were used for normally distributed data and medians for data not meeting this assumption. Fisher's exact and χ2 tests were used to evaluate the associations between variables and Internet use. RESULTS: A total of 282 surveys were analyzed. The majority of participants, 83.3%, were white. The mean age was 55.8 years old. Referrals to urogynecology practices were most frequently from obstetrician/gynecologists (39.9%) and primary care providers (27.8%). Subjects were well distributed geographically, with the largest representation from the South (38.0%). Almost one third (29.9%) were most bothered by prolapse complaints, 22.0% by urgency urinary incontinence, 20.9% by stress urinary incontinence, 14.9% by urgency/frequency symptoms, and 4.1% by fecal incontinence. The majority, 75.0%, described high Internet use, whereas 8.5% moderately and 4.8% minimally used the Internet. Women most often used the Internet for personal motivations including medical research (76.4%), and 42.6% reported Google to be their primary search engine. Despite this, only 4.9% primarily used the Internet to learn about their pelvic floor condition, more commonly consulting an obstetrician-gynecologist for this information (39.4%). The majority (74.1%) held a social networking account, and 45.9% visited these daily. Nearly half, 41.7%, expressed the desire to use social networking websites to learn about their condition. Women <65 years old were significantly more likely to have high Internet use (83.4% vs 68.8%, P = .018) and to desire using social networking websites to learn about their pelvic floor complaint (P = .008). The presenting complaint was not associated with Internet use (P = .905) or the desire to use social networking websites to learn about pelvic floor disorders (P = .201). CONCLUSION: Women presenting to urogynecology practices have high Internet use and a desire to learn about their conditions via social networking websites. Despite this, obstetrician-gynecologists remain a common resource for information. Nonetheless, urogynecology practices and national organizations would likely benefit from increasing their Internet resources for patient education in pelvic floor disorders, though patients should be made aware of available resources.


Assuntos
Informação de Saúde ao Consumidor/estatística & dados numéricos , Internet/estatística & dados numéricos , Distúrbios do Assoalho Pélvico , Rede Social , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos Transversais , Feminino , Humanos , Pessoa de Meia-Idade , Distúrbios do Assoalho Pélvico/cirurgia , Ferramenta de Busca/estatística & dados numéricos , Inquéritos e Questionários , Estados Unidos , Adulto Jovem
9.
Int Urogynecol J ; 27(11): 1705-1711, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27116196

RESUMO

INTRODUCTION AND HYPOTHESIS: Mindfulness-based stress reduction (MBSR) is a standardized meditation program that may be an effective therapy for interstitial cystitis/bladder pain syndrome (IC/BPS), a condition exacerbated by stress. The aims of this study were to explore whether MBSR improved IC/BPS symptoms and the feasibility/acceptability of MSBR among women with IC/BPS. METHODS: This randomized controlled trial included women with IC/BPS undergoing first- or second-line therapies. Women were randomized to continuation of usual care (UC) or an 8-week MBSR class + usual care (MBSR). Participants completed baseline and 8-week post-treatment questionnaires, including the O'Leary-Sant Symptom Problem Index (OSPI), the visual analog pain scale (VAS), the Short Form Health Survey (SF-12), the Female Sexual Function Index (FSFI), and the Pain Self-Efficacy Questionnaire (PSEQ). The Global Response Assessment (GRA) was completed post-treatment. Analyses were performed using Student's t test, Chi-squared, and MANOVA where appropriate. RESULTS: Eleven women were randomized to UC and 9 to MBSR, without differences in group characteristics. More MBSR participants' symptoms were improved on the GRA (7 out of 8 [87.5 %] vs 4 out of 11 [36.4 %], p = 0.03). The MBSR group showed greater improvement in the OSPI total (p = 0.0498) and problem scores (p = 0.036); the OSPI symptom score change did not differ. PSEQ scores improved in MBSR compared with UC (p = 0.035). VAS, SF-12, and FSFI change did not differ between groups. Eighty-six percent of MBSR participants felt more empowered to control symptoms, and all participants planned to continue MBSR. CONCLUSIONS: This trial provides initial evidence that MBSR is a promising adjunctive therapy for IC/BPS. Its benefit may arise from patients' empowerment and ability to cope with symptoms.


Assuntos
Cistite Intersticial/psicologia , Atenção Plena/métodos , Estresse Psicológico/terapia , Adulto , Análise de Variância , Distribuição de Qui-Quadrado , Feminino , Humanos , Pessoa de Meia-Idade , Qualidade de Vida , Inquéritos e Questionários , Resultado do Tratamento
10.
Int Urogynecol J ; 26(12): 1797-801, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26142348

RESUMO

INTRODUCTION AND HYPOTHESIS: Perineorrhaphy is performed for the prevention of recurrent prolapse, improved sexual function, treatment of pain, and cosmesis. Its use is based on expert opinion with few objective data. We aimed to describe factors that are important to surgeons when deciding to perform perineorrhaphy and variations in surgical technique. METHODS: We administered an anonymous survey to surgeon attendees at the 2014 SGS annual scientific meeting regarding which factors are important when deciding to perform a perineorrhaphy and details of their surgical technique. Surgeons rated the importance of factors; one-way ANOVA was used to rank the decision factors and post hoc pairwise comparisons with Fisher's least significant difference method were used to evaluate the importance between factors. RESULTS: A total of 183 out of 360 surgeon attendees responded. Most were between ages 36 and 60 (79 %), 56 % were female, 64 % practiced in an academic environment, and 64 % had undergone subspecialty training. An enlarged genital hiatus (GH) ranked as the most important factor influencing the decision to perform a perineorrhaphy, followed by a concomitant prolapse procedure (p < 0.001). Sexual function and cosmesis were rated as being less important. The decision to perform perineorrhaphy was made with the patient in 65 % of cases, and otherwise in the operating room. Significant heterogeneity exists regarding surgeon suture preference and how muscles were re-approximated. Most (81 %) reported incorporating structures both proximal and distal to the hymen in their repairs. CONCLUSION: Genital hiatus size and concomitant prolapse procedures ranked highest in surgeons' decision to perform a perineorrhaphy. Significant heterogeneity exists in the indications for and technique used to perform perineorrhaphy.


Assuntos
Atitude do Pessoal de Saúde , Ginecologia , Prolapso de Órgão Pélvico/cirurgia , Períneo/cirurgia , Adulto , Tomada de Decisão Clínica , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Seleção de Pacientes , Prolapso de Órgão Pélvico/diagnóstico , Prolapso de Órgão Pélvico/etiologia , Padrões de Prática Médica
11.
Opt Express ; 22(23): 27942-57, 2014 Nov 17.
Artigo em Inglês | MEDLINE | ID: mdl-25402035

RESUMO

We describe a technique for dynamic quantum optical arbitrary-waveform generation and manipulation, which is capable of mode selectively operating on quantum signals without inducing significant loss or decoherence. It is built upon combining the developed tools of quantum frequency conversion and optical arbitrary waveform generation. Considering realistic parameters, we propose and analyze applications such as programmable reshaping of picosecond-scale temporal modes, selective frequency conversion of any one or superposition of those modes, and mode-resolved photon counting. We also report on experimental progress to distinguish two overlapping, orthogonal temporal modes, demonstrating over 8 dB extinction between picosecond-scale time-frequency modes, which agrees well with our theory. Our theoretical and experimental progress, as a whole, points to an enabling optical technique for various applications such as ultradense quantum coding, unity-efficiency cavity-atom quantum memories, and high-speed quantum computing.


Assuntos
Óptica e Fotônica/instrumentação , Teoria Quântica , Desenho de Equipamento , Fótons
12.
Obstet Gynecol ; 142(3): 555-570, 2023 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-37543737

RESUMO

OBJECTIVE: To systematically review the literature and provide clinical practice guidelines regarding various nonestrogen therapies for treatment of genitourinary syndrome of menopause (GSM). DATA SOURCES: MEDLINE, EMBASE, ClinicalTrials.gov , and Cochrane databases were searched from inception to July 2021. We included comparative and noncomparative studies. Interventions and comparators were limited to seven products that are commercially available and currently in use (vaginal dehydroepiandrosterone [DHEA], ospemifene, laser or energy-based therapies, polycarbophil-based vaginal moisturizer, Tibolone, vaginal hyaluronic acid, testosterone). Topical estrogen, placebo, other nonestrogen products, as well as no treatment were considered as comparators. METHODS OF STUDY SELECTION: We double-screened 9,131 abstracts and identified 136 studies that met our criteria. Studies were assessed for quality and strength of evidence by the systematic review group. TABULATION, INTEGRATION, AND RESULTS: Information regarding the participants, details on the intervention and comparator and outcomes were extracted from the eligible studies. Alternative therapies were similar or superior to estrogen or placebo with minimal increase in adverse events. Dose response was noted with vaginal DHEA and testosterone. Vaginal DHEA, ospemifene, erbium and fractional carbon dioxide (CO 2 ) laser, polycarbophil-based vaginal moisturizer, tibolone, hyaluronic acid, and testosterone all improved subjective and objective signs of atrophy. Vaginal DHEA, ospemifene, tibolone, fractional CO 2 laser, polycarbophil-based vaginal moisturizer, and testosterone improved sexual function. CONCLUSION: Most nonestrogen therapies are effective treatments for the various symptoms of GSM. There are insufficient data to compare nonestrogen options to each other.


Assuntos
Ácido Hialurônico , Menopausa , Feminino , Humanos , Ácido Hialurônico/uso terapêutico , Ácido Hialurônico/farmacologia , Vagina , Estrogênios/uso terapêutico , Testosterona/farmacologia , Desidroepiandrosterona/uso terapêutico , Desidroepiandrosterona/efeitos adversos
13.
Int Urogynecol J ; 27(6): 977-8, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26879086
14.
Opt Express ; 10(3): 177-82, 2002 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-19424347

RESUMO

We report on traveling-wave quadrature squeezing generated in an integrated optical circuit fabricated with periodicallypoled lithium niobate waveguides. The device integrates a secondharmonic-generation stage, waveguide couplers, spatial mode filters, and a degenerate-optical-parametric-amplification stage. The integrationpromises to create a low power, compact, and simple source of wideband squeezed light. -1 dB of squeezing is directly measured using 20 ps pulses with peak powers near 6W.

15.
Biotechnol Prog ; 20(6): 1689-96, 2004.
Artigo em Inglês | MEDLINE | ID: mdl-15575700

RESUMO

Granulocyte-macrophage colony-stimulating factor (GM-CSF) is an important cytokine in the mammalian immune system. It has been expressed in Escherichia coli with the same biological activity as the native protein. Here, we report the synthesis of a murine recombinant GM-CSF in an E. coli cell-free protein synthesis system with a high yield. Since there are two disulfide bonds in the native structure of GM-CSF, an oxidizing redox potential of the reaction mixture was required. By pretreating the cell extract with iodoacetamide (IAM), the reducing activity of the cell extract was inactivated, and upon further application of an oxidized glutathione buffer, most of the synthesized GM-CSF was found in its oxidized form. However, the GM-CSF thus formed showed low activity because of poor folding. With the addition of DsbC, the periplasmic disulfide isomerase from E. coli, a high yield of active GM-CSF was produced in the cell-free reaction. Finally, successful folding of the cell-free synthesized GM-CSF-his6 was confirmed by its cell-proliferation activity after purification with a Ni2+ chelating column.


Assuntos
Escherichia coli/genética , Escherichia coli/metabolismo , Fator Estimulador de Colônias de Granulócitos/biossíntese , Fator Estimulador de Colônias de Granulócitos/química , Engenharia de Proteínas/métodos , Animais , Sistema Livre de Células , Regulação Bacteriana da Expressão Gênica/fisiologia , Melhoramento Genético/métodos , Camundongos , Proteínas Recombinantes
16.
Blood ; 109(8): 3393-9, 2007 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-17164345

RESUMO

The unique immunoglobulin (Ig) idiotype on the surface of each B-cell lymphoma represents an ideal tumor-specific antigen for use as a therapeutic vaccine. We have used an Escherichia coli-based, cell-free protein-expression system to produce a vaccine within hours of cloning the Ig genes from a B-cell tumor. We demonstrated that a fusion protein consisting of an idiotypic single chain Fv antibody fragment (scFv) linked to a cytokine (GM-CSF) or to an immunostimulatory peptide was an effective lymphoma vaccine. These vaccines elicited humoral immune responses against the native Ig protein displayed on the surface of a tumor and protected mice against tumor challenge with efficacy equal to that of the conventional Ig produced in a mammalian cell and chemically coupled to keyhole limpet hemocyanin. The cell-free E coli system offers a platform for rapidly generating individualized vaccines, thereby allowing much more efficient application in the clinic.


Assuntos
Anticorpos Monoclonais/biossíntese , Antígenos de Neoplasias/biossíntese , Vacinas Anticâncer/biossíntese , Fator Estimulador de Colônias de Granulócitos e Macrófagos/biossíntese , Idiótipos de Imunoglobulinas/biossíntese , Região Variável de Imunoglobulina/biossíntese , Linfoma de Células B/terapia , Proteínas Recombinantes de Fusão/biossíntese , Animais , Anticorpos Monoclonais/genética , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/uso terapêutico , Antígenos de Neoplasias/genética , Antígenos de Neoplasias/imunologia , Antígenos de Neoplasias/efeitos da radiação , Vacinas Anticâncer/genética , Vacinas Anticâncer/imunologia , Vacinas Anticâncer/uso terapêutico , Fator Estimulador de Colônias de Granulócitos e Macrófagos/genética , Fator Estimulador de Colônias de Granulócitos e Macrófagos/imunologia , Humanos , Idiótipos de Imunoglobulinas/genética , Idiótipos de Imunoglobulinas/imunologia , Idiótipos de Imunoglobulinas/uso terapêutico , Região Variável de Imunoglobulina/genética , Região Variável de Imunoglobulina/imunologia , Região Variável de Imunoglobulina/uso terapêutico , Linfoma de Células B/genética , Linfoma de Células B/imunologia , Camundongos , Neoplasias Experimentais/genética , Neoplasias Experimentais/imunologia , Neoplasias Experimentais/terapia , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia
17.
Biotechnol Bioeng ; 89(5): 503-11, 2005 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-15669088

RESUMO

The idiotype (Id)-granulocyte-macrophage colony-stimulating factor (GM-CSF) fusion proteins are potential vaccines for immunotherapy of B-cell lymphoma. In this study, four vaccine candidates were constructed by fusing murine GM-CSF to the amino- or carboxy-terminus of the 38C13 murine B-lymphocyte Id scFv with two different arrangements of the variable regions of the heavy chain and light chain (VL-VH and VH-VL). scFv (VH-VL) and GM-CSF/scFv fusion proteins were expressed in an Escherichia coli cell-free protein synthesis system. In order to promote disulfide bond formation during cell-free expression, cell extract was pretreated with iodoacetamide (IAM), and a sulfhydryl redox buffer composed of oxidized and reduced glutathione was added. The E. coli periplasmic disulfide isomerase, DsbC, was also added to rearrange incorrectly formed disulfide linkages. The 38C13 B-lymphocyte Id scFv was expressed with 30% of its soluble yield in active form (43 microg/ml) when tested with an anti-idiotypic mAb, S1C5, as the capture antibody in radioimmunoassay. It was found that the amino-terminal GM-CSF fusion proteins, GM-VL-VH and GM-VH-VL, showed much higher activity than the carboxy-terminal GM-CSF fusion proteins, VL-VH-GM and VH-VL-GM, in stimulating the cell proliferation of a GM-CSF-dependent cell line, NFS-60. Between the two amino-terminal GM-CSF fusion proteins, GM-VL-VH showed a higher total and soluble yield than GM-VH-VL.


Assuntos
Linfoma de Células B/terapia , Proteínas/metabolismo , Vacinas/química , Animais , Anticorpos Monoclonais/metabolismo , Autorradiografia , Vacinas Anticâncer/imunologia , Proliferação de Células , Sistema Livre de Células , Dissulfetos/química , Escherichia coli/enzimologia , Escherichia coli/genética , Regulação Bacteriana da Expressão Gênica , Fator Estimulador de Colônias de Granulócitos e Macrófagos/química , Fator Estimulador de Colônias de Granulócitos e Macrófagos/genética , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Cadeias Pesadas de Imunoglobulinas , Idiótipos de Imunoglobulinas , Cadeias Leves de Imunoglobulina , Região Variável de Imunoglobulina , Camundongos , Isomerases de Dissulfetos de Proteínas/metabolismo , Dobramento de Proteína , Estrutura Terciária de Proteína , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/metabolismo , Vacinas/genética , Vacinas/imunologia
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