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  • 1
    In: Anesthesiology, Ovid Technologies (Wolters Kluwer Health), Vol. 93, No. 3A ( 2000-09-01), p. A-609
    Type of Medium: Online Resource
    ISSN: 0003-3022
    RVK:
    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2000
    detail.hit.zdb_id: 2016092-6
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  • 2
    In: Journal of Molecular and Cellular Cardiology, Elsevier BV, Vol. 34, No. 3 ( 2002-03), p. 349-359
    Type of Medium: Online Resource
    ISSN: 0022-2828
    Language: English
    Publisher: Elsevier BV
    Publication Date: 2002
    detail.hit.zdb_id: 1469767-1
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  • 3
    Online Resource
    Online Resource
    SAGE Publications ; 2022
    In:  Annals of Otology, Rhinology & Laryngology Vol. 131, No. 12 ( 2022-12), p. 1346-1352
    In: Annals of Otology, Rhinology & Laryngology, SAGE Publications, Vol. 131, No. 12 ( 2022-12), p. 1346-1352
    Abstract: Manual jet ventilation is a specialized oxygenation and ventilation technique that is not available in all facilities due to lack of technical familiarity and fear of complications. The objective is to review our center’s 15 year experience with low pressure low frequency jet ventilation (LPLFJV). Methods: Retrospective review of procedures utilizing LPLFJV from 2005 to 2019 were performed collecting patient demographic, surgery type and complications. Fisher exact test, Chi square, and t-test were used to determine statistical significance. Results: Four hundred fifty-seven patients underwent a total of 891 microlaryngeal surgeries—279 cases for voice disorders, 179 for lesions, and 433 for airway stenosis. The peak jet pressure for all cases did not exceed 20 psi and average peak pressure for the last 100 procedures in this case series was 14.9 ± 4.6 psi. The average lowest oxygen saturation for all cases was 95% ± 0.6%. Brief intubation was required in 154 cases (17%). Surgical duration was significantly longer for cases requiring intubation P  〈  .001. The need for intubation was not associated with smoking or cardiopulmonary disease, but was strongly associated with body mass index (BMI). Intubation rates were 7% for normal weight (BMI  〈  25, N = 216), 13% for overweight (BMI 25-30, N = 282), 24% for obese (BMI 30-40, N = 342), and 37% for morbidly obese (BMI  〉  40, N = 52) patients. Three patients developed respiratory distress in the recovery unit and 2 patients required intubation. Conclusion: LPLFJV assisted by intermittent endotracheal intubation is an exceedingly safe and effective intraoperative oxygenation and ventilationmodality for a broad variety of laryngeal procedure.
    Type of Medium: Online Resource
    ISSN: 0003-4894 , 1943-572X
    Language: English
    Publisher: SAGE Publications
    Publication Date: 2022
    detail.hit.zdb_id: 2033055-8
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  • 4
    In: Journal of Neurochemistry, Wiley, Vol. 113, No. 3 ( 2010-05), p. 637-648
    Type of Medium: Online Resource
    ISSN: 0022-3042 , 1471-4159
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2010
    detail.hit.zdb_id: 2020528-4
    SSG: 12
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  • 5
    Online Resource
    Online Resource
    American Society of Hematology ; 2013
    In:  Blood Vol. 122, No. 21 ( 2013-11-15), p. 969-969
    In: Blood, American Society of Hematology, Vol. 122, No. 21 ( 2013-11-15), p. 969-969
    Abstract: It is well established that sickle cell disease (SCD) manifests global perturbations of hemostasis. Vaso-occlusion, inflammation and coagulopathy all likely contribute to the protean complications of SCD. Central to both inflammation and coagulation is the monocyte. These cells can be profoundly pro-inflammatory and can express tissue factor on their surface and thus influence both inflammation and coagulation. Monocytosis is common in SCD, as is steady state monocyte activation. Exaggerated monocytic response to stimulus may also contribute to the severity of acute events. Thus, agents that regulate monocyte function are potentially of significant relevance in SCD. To this end, we have discovered that the heat shock proteins (HSPs) are potential master regulators of these cells. We previously demonstrated that inhibition of one such HSP, HSP90, could completely ablate the profound and hyper-responsive monocytic inflammatory release upon lipopolysacchiride (LPS) challenge. Inhibition of HSP90 also blocked LPS induced NFk-B translocation to the nucleus. Thus, these results suggested a potent role for HSP90 in LPS-induced monocyte based inflammation. However, the role of HSP90 is cytokine-induced monocyte activation was speculative. The role of HSP90 in monocyte tissue factor expression, or reactive oxygen species (ROS) generation remained unknown. Objectives We sought to determine the role of HSP90 in regulating the pro-inflammatory, pro-coagulant, and ROS generating potential in monocytes. We then wanted to establish a potential role for HSP90 inhibition. Methods Peripheral blood mononuclear cells (PBMCs) were isolated from whole blood using Ficoll density separation. Flow cytometry was employed to measure the LPS- or cytokine-induced monocyte tissue factor expression and ROS generation. Monocyte ROS generation was detected using L012 based chemiluminescence or visualized in individual cells using flow cytometry with CELLROX. Inflammatory cytokines and tissue factor gene expression was evaluated using quantitative real-time polymerase chain reaction (qRT-PCR). NFkB translocation to the nucleus was detected via cell fractionation followed by western blotting or indirect immunofluorescence. PBMCs or THP-1 cells were pre-treated with the HSP90 inhibitors 17-DMAG or AUY922 prior to assay. Monocyte- induced endothelial cell permeability was measured using endothelial cell-substrate impedance sensing (ECIS). Unless otherwise stated p 〈 0.05. Results At baseline, PBMCs from patients with SCD demonstrated elevated monocyte ROS generation and tissue factor expression than those from healthy controls. Inhibition of significantly reduced these measures of steady state monocyte activation. HSP90 inhibition also inhibited both LPS and cytokine induced tissue mRNA accumulation and subsequent cell surface expression of tissue factor in monocytes. Cytokine- induced ROS generation was significantly interrupted in monocytes upon inhibition of HSP90. A panel of monocyte pro-inflammatory genes could be inhibited with AUY922, whereas the anti-inflammatory cytokine, IL-10, was induced upon HSP90 inhibition with AUY922. Mechanistically, we also noted a profound translocation of NFkB to the monocyte nucleus upon cytokine stimulation. Consistent with the effects on tissue factor expression, pro-inflammatory potential, and ROS generation, this translocation could be completely ablated with HSP90 inhibition. Importantly, HSP90 inhibitors significantly attenuated the LPS-activated monocyte -induced lung microvascular endothelial permeability. Conclusion Our data suggest that Hsp90 inhibitors significantly reduced the both pro-inflammatory and pro-coagulatory potential of PBMCs from patients with SCD. These results thus position HSP90 as a potential master regulator of hemostasis, endothelial cell permeability, and thus suggest that HSP90 is an attractive therapeutic target in patients with SCD. Disclosures: Kutlar: Celgene Corporation: Research Funding. Meiler:Celgene Corporation: Research Funding.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    RVK:
    RVK:
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2013
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
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  • 6
    In: The FASEB Journal, Wiley, Vol. 27, No. S1 ( 2013-04)
    Type of Medium: Online Resource
    ISSN: 0892-6638 , 1530-6860
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2013
    detail.hit.zdb_id: 1468876-1
    SSG: 12
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  • 7
    Online Resource
    Online Resource
    Ovid Technologies (Wolters Kluwer Health) ; 2011
    In:  Journal of Pediatric Hematology/Oncology Vol. 33, No. 8 ( 2011-12), p. 589-591
    In: Journal of Pediatric Hematology/Oncology, Ovid Technologies (Wolters Kluwer Health), Vol. 33, No. 8 ( 2011-12), p. 589-591
    Type of Medium: Online Resource
    ISSN: 1077-4114
    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2011
    detail.hit.zdb_id: 2047125-7
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  • 8
    Online Resource
    Online Resource
    Springer Science and Business Media LLC ; 2002
    In:  Nature Genetics Vol. 30, No. 2 ( 2002-2), p. 205-209
    In: Nature Genetics, Springer Science and Business Media LLC, Vol. 30, No. 2 ( 2002-2), p. 205-209
    Type of Medium: Online Resource
    ISSN: 1061-4036 , 1546-1718
    RVK:
    Language: English
    Publisher: Springer Science and Business Media LLC
    Publication Date: 2002
    detail.hit.zdb_id: 1494946-5
    SSG: 12
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  • 9
    In: Blood, American Society of Hematology, Vol. 120, No. 21 ( 2012-11-16), p. 1007-1007
    Abstract: Abstract 1007 Pain is a common and serious complication of sickle cell disease (SCD), which is frequently disabling and difficult to treat. Acute painful crises account for the vast majority of healthcare encounters ( 〉 90% of hospitalizations), and many patients with SCD suffer from chronic pain. Despite several well designed clinical studies, which have brought to light the high incidence and severity of pain in this patient population, sickle cell pain remains an understudied, undermanaged, and poorly understood condition. Previous work has largely relied on self-reporting instruments, such as carefully designed pain diaries to study the epidemiology of pain in this patient cohort. In contrast, very little is known about the usefulness of methods that measure pain in response to a standardized stimulus and the correlation of experimentally induced pain to more traditional instruments such as pain diaries and opioid use. In this study we explored the use of pressure algometry to measure pain thresholds in adult patients with SCD. Our study enrolled 167 adult patients with SCD [163 SS, 1 SD Los Angeles, 3 Sβ0-thal; 108 on Hydroxyurea (HU), 59 off HU; 83 females and 84 males; mean age 31 years (range 16–61)] and forty racially matched controls (23 females and 17 males; mean age 35 years (range 17–61). The pain threshold, defined as the lowest pressure at which pain is induced, was measured by pressure algometry in three anatomic muscle groups (masseter, trapezius, and ulnar). Four measurements were obtained at each site and averaged for analysis. 118 patients were tested with a manual algometer (Wagner FDIX™, Wagner Instruments, Greenwich, CT) and 49 patients with a computerized model (Algomed, Medoc Ltd., Israel). The data sets for the two algometer groups were highly correlated for each anatomic site and were therefore combined for the final analysis (Pearson's correlation coefficients: masseter readings, r=0.78, p=2.73E-09; trapezius readings, r=0.85, p=4.28E-12; ulnar readings, r=0.68, p=3.26E-06). In addition, self-reported pain and distress were monitored prospectively for six months using a validated pain diary (ordinal scales of 0–9, respectively; J Natl Med Assoc. 2005 February; 97(2): 183–193). Opioid use, expressed as morphine equivalents, and the frequency of vaso-occlusive events are being recorded prospectively for twelve months. Algometer readings were compared using a linear model with age and gender as covariates. Data are reported as mean ± SEM. A p-value of 〈 0.05 is significant. Adult patients with SCD experienced pain at significantly lower pressures at all anatomic sites tested compared to racially matched controls. Pain threshold measurements in the ulnar muscle group achieved the best discrimination between sickle cell and control subjects. (Patients vs. controls (KPa): Masseter: 150± 4.35 vs. 193± 13.24, p= 1.99E–05; trapezius: 265±10.23 vs. 383± 37.99, p= 2.78E–05; ulnar: 371±10.26 vs. 518±34.59, p= 6.28E–09). Ulnar algometer readings also correlated with self-reported pain scores (r=-0.226, p=0.0097). There was no correlation between pain threshold measurements and self-reported distress or the use of hydroxyurea. Quantitative sensory testing revealed that adult patients with SCD are hyperalgesic in response to a standardized pressure stimulus. Of the three anatomic sites tested, ulnar pain threshold readings produced the strongest separation between sickle cell and control subjects and correlated with self-reported pain. Quantitative sensory testing may provide a useful research and clinical tool to study the biological mechanisms of pain in SCD and the therapeutic efficacy of psychosocial and pharmacological interventions. Disclosures: Fillingim: Algynomics: Consultancy, Shareholder Other.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    RVK:
    RVK:
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2012
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
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  • 10
    In: Blood, American Society of Hematology, Vol. 118, No. 21 ( 2011-11-18), p. 1071-1071
    Abstract: Abstract 1071 Zinc finger nucleases (ZFNs) are custom-designed DNA binding proteins that produce DNA double-strand breaks (DSBs) at predetermined genomic sites, stimulating homology-directed repair in the presence of donor template by many orders of magnitude over the spontaneous rate. The ability to target specific genes with ZFN technology opens therapeutic opportunities for gene correction and selective gene silencing. Sickle cell anemia (SCA) is an ideal disease target because correction of the single gene β-globin mutation in patient-derived, autologous hematopoietic stem progenitor cells (HSPCs) promises to be curative. One of the barriers to ZFN-based gene correction is the lack of a nonviral delivery system that achieves bulk transport of the nucleases to hard-to-transfect target cells, such as embryonic and HSPCs. To address this challenge, we set out to develop a delivery platform that is (i) gentle to the cell, (ii) provides tunable delivery rates, and (iii) achieves improved spatio-temporal control of the nucleases. To reconcile these goals, we have explored a method for direct delivery of ZFNs as proteins by receptor-mediated endocytosis. We selected the transferrin receptor pathway as our lead candidate on the rationale that all nucleated cells, including HSPCs, must import elemental iron to remain viable under ex vivo culture conditions. To test the feasibility of this strategy, this initial work used a ZFN pair targeted against a model GFP transgene. We optimized expression by pilot scale fermentation in an Escherichia coli host-vector system and purification to homogeneity by serial chromatography. We conjugated the purified ZFNs to the iron carrier protein, transferrin (tf), using SPDP, an amine and sulfhydryl reactive heterobifunctional crosslinker. The resulting disulfide linkage is designed to undergo scission (“self-immolation”) upon entry into the intracellular reducing environment. In vitro DNA cleavage assays and surface plasmon resonance binding assays demonstrated that ZFNs remained competent for target sequence cleavage following conjugation, with only mild to quantitative impairment of activity. To analyze delivery in biological systems, we measured time- and dose-dependence of tf-mediated ZFN uptake in human osteosarcoma (U2OS 2–6–3) cells. ZFNs in DAPI stained cell nuclei were detected by indirect immunofluorescence and signal intensity was measured in projections of deconvolved depth coded z-stacks. Nuclear uptake of tf-ZFN protein occurred in 〉 95% of cells, was dose-dependent and linear with time in the lower dose ranges, and reached saturation as early as 60 min. Importantly, maximal nuclear uptake was indistinguishable from ZFN plasmid treated cells. These results indicate that endocytic delivery of ZFNs readily traverses the cellular membrane, overcomes the potential hurdle of endosomal trapping, and targets the nucleus with high efficiency. To demonstrate gene targeting activity, we used the U2OS 2–6–3 cell assay which bears a tandem transgene array at a single locus that is cleavable by our GFP ZFNs. Cells were transfected with lacI-ECFP to mark the target locus, incubated with tf-ZFNs, fixed, and stained for 53BP1, a signaling protein that marks DSBs. Recruitment of 53BP1 to the target locus was observed in 13% (18/135) of tf-ZFN treated cells, whereas no recruitment (0/152) was observed in untreated cells. These findings demonstrate that tf-conjugated ZFNs retain cleavage activity after nuclear uptake in a significant percentage of cells. To determine whether the tf-ZFNs are capable of stimulating gene correction, we transfected primary mouse adult fibroblasts carrying a mutant GFP transgene with donor template, incubated with tf-ZFNs, and evaluated cells at 72 h for gene correction as evidenced by GFP expression. Flow cytometry revealed a gene correction rate of 1–2%, identical to ZFN plasmid transfected cells, demonstrating that the technology of shuttling ZFN proteins to the cell interior via the tf-receptor pathway can deliver bioactive ZFNs to the nuclear compartment, target specific gene sequences, and induce homology-directed repair in the presence of donor DNA. We are currently testing these methods in hematopoietic stem cells, with the ultimate goal of correcting the sickle globin allele. Toward this end, we plan to adapt these approaches for high-throughput transfer of ZFN proteins directly to the hematopoietic stem progenitor cell. Disclosures: No relevant conflicts of interest to declare.
    Type of Medium: Online Resource
    ISSN: 0006-4971 , 1528-0020
    RVK:
    RVK:
    Language: English
    Publisher: American Society of Hematology
    Publication Date: 2011
    detail.hit.zdb_id: 1468538-3
    detail.hit.zdb_id: 80069-7
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