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Uruguay Oncology Collection
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1.
Pediatr Res ; 95(1): 93-101, 2024 Jan.
Article in English | MEDLINE | ID: mdl-37087539

ABSTRACT

BACKGROUND: Clinical translation of the extracorporeal artificial placenta (AP) is impeded by the high risk for intracranial hemorrhage in extremely premature newborns. The Nitric Oxide Surface Anticoagulation (NOSA) system is a novel non-thrombogenic extracorporeal circuit. This study aims to test the NOSA system in the AP without systemic anticoagulation. METHODS: Ten extremely premature lambs were delivered and connected to the AP. For the NOSA group, the circuit was coated with DBHD-N2O2/argatroban, 100 ppm nitric oxide was blended into the sweep gas, and no systemic anticoagulation was given. For the Heparin control group, a non-coated circuit was used and systemic anticoagulation was administered. RESULTS: Animals survived 6.8 ± 0.6 days with normal hemodynamics and gas exchange. Neither group had any hemorrhagic or thrombotic complications. ACT (194 ± 53 vs. 261 ± 86 s; p < 0.001) and aPTT (39 ± 7 vs. 69 ± 23 s; p < 0.001) were significantly lower in the NOSA group than the Heparin group. Platelet and leukocyte activation did not differ significantly from baseline in the NOSA group. Methemoglobin was 3.2 ± 1.1% in the NOSA group compared to 1.6 ± 0.6% in the Heparin group (p < 0.001). CONCLUSIONS: The AP with the NOSA system successfully supported extremely premature lambs for 7 days without significant bleeding or thrombosis. IMPACT: The Nitric Oxide Surface Anticoagulation (NOSA) system provides effective circuit-based anticoagulation in a fetal sheep model of the extracorporeal artificial placenta (AP) for 7 days. The NOSA system is the first non-thrombogenic circuit to consistently obviate the need for systemic anticoagulation in an extracorporeal circuit for up to 7 days. The NOSA system may allow the AP to be implemented clinically without systemic anticoagulation, thus greatly reducing the intracranial hemorrhage risk for extremely low gestational age newborns. The NOSA system could potentially be applied to any form of extracorporeal life support to reduce or avoid systemic anticoagulation.


Subject(s)
Extracorporeal Membrane Oxygenation , Premature Birth , Thrombosis , Pregnancy , Humans , Female , Sheep , Animals , Nitric Oxide , Placenta/physiology , Heparin , Hemorrhage/complications , Thrombosis/prevention & control , Anticoagulants/pharmacology , Intracranial Hemorrhages/complications
2.
Perfusion ; : 2676591241240725, 2024 Mar 22.
Article in English | MEDLINE | ID: mdl-38519444

ABSTRACT

INTRODUCTION: A radical paradigm shift in the treatment of premature infants failing conventional treatment is to recreate fetal physiology using an extracorporeal Artificial Placenta (AP). The aim of this study is to evaluate the effects of changing fetal hemoglobin percent (HbF%) on physiology and circuit function during AP support in an ovine model. METHODS: Extremely premature lambs (n = 5) were delivered by cesarean section at 117-121 d estimated gestational age (EGA) (term = 145d), weighing 2.5 ± 0.35 kg. Lambs were cannulated using 10-14Fr cannulae for drainage via the right jugular vein and reinfusion via the umbilical vein. Lambs were intubated and lungs were filled with perfluorodecalin to a meniscus with a pressure of 5-8 cm H2O. The first option for transfusion was fetal whole blood from twins followed by maternal red blood cells. Arterial blood gases were used to titrate AP support to maintain fetal blood gas values. RESULTS: The mean survival time on circuit was 119.6 ± 39.5 h. Hemodynamic parameters and lactate were stable throughout. As more adult blood transfusions were given to maintain hemoglobin at 10 mg/dL, the HbF% declined, reaching 40% by post operative day 7. The HbF% was inversely proportional to flow rates as higher flows were required to maintain adequate oxygen saturation and perfusion. CONCLUSIONS: Transfusion of adult blood led to decreased fetal hemoglobin concentration during AP support. The HbF% was inversely proportional to flow rates. Future directions include strategies to decrease the priming volume and establishing a fetal blood bank to have blood rich in HbF.

3.
Breast Cancer Res Treat ; 197(2): 277-285, 2023 Jan.
Article in English | MEDLINE | ID: mdl-36380012

ABSTRACT

PURPOSE: Breast cancer risk is elevated in pathogenic germline BRCA 1/2 mutation carriers due to compromised DNA quality control. We hypothesized that if immunosurveillance promotes tumor suppression, then normal/benign breast lobules from BRCA carriers may demonstrate higher immune cell densities. METHODS: We assessed immune cell composition in normal/benign breast lobules from age-matched women with progressively increased breast cancer risk, including (1) low risk: 19 women who donated normal breast tissue to the Komen Tissue Bank (KTB) at Indiana University Simon Cancer Center, (2) intermediate risk: 15 women with biopsy-identified benign breast disease (BBD), and (3) high risk: 19 prophylactic mastectomies from women with germline mutations in BRCA1/2 genes. We performed immunohistochemical stains and analysis to quantitate immune cell densities from digital images in up to 10 representative lobules per sample. Median cell counts per mm2 were compared between groups using Wilcoxon rank-sum tests. RESULTS: Normal/benign breast lobules from BRCA carriers had significantly higher densities of immune cells/mm2 compared to KTB normal donors (all p < 0.001): CD8 + 354.4 vs 150.9; CD4 + 116.3 vs 17.7; CD68 + 237.5 vs 57.8; and CD11c + (3.5% vs 0.4% pixels positive). BBD tissues differed from BRCA carriers only in CD8 + cells but had higher densities of CD4 + , CD11c + , and CD68 + immune cells compared to KTB donors. CONCLUSIONS: These preliminary analyses show that normal/benign breast lobules of BRCA mutation carriers contain increased immune cells compared with normal donor breast tissues, and BBD tissues appear overall more similar to BRCA carriers.


Subject(s)
Breast Neoplasms , Female , Humans , Breast Neoplasms/pathology , Breast/pathology , Germ-Line Mutation , Genes, BRCA1 , CD8-Positive T-Lymphocytes/pathology , Mutation , BRCA1 Protein/genetics
4.
BMC Anesthesiol ; 23(1): 415, 2023 12 18.
Article in English | MEDLINE | ID: mdl-38110877

ABSTRACT

BACKGROUND: Intraoperative pulmonary embolism (PE) with cardiac arrest (CA) represents a critical and potentially fatal condition. Available treatments include systemic thrombolysis, catheter-based thrombus fragmentation or aspiration, and surgical embolectomy. However, limited studies are focused on the optimal treatment choice for this critical condition. We present a case series and an updated review of the management of intraoperative CA secondary to PE. METHODS: A retrospective review of patients who developed high-risk intraoperative PE was performed between June 2012 and June 2022. For the updated review, a literature search on PubMed and Scopus was conducted which resulted in the inclusion of a total of 46 articles. RESULTS: A total of 196 174 major non-cardiac surgeries were performed between 2012 and 2022. Eight cases of intraoperative CA secondary to high-risk PE were identified. We found a mortality rate of 75%. Anticoagulation therapy was administered to one patient (12.5%), while two patients (25%) underwent thrombolysis, and one case (12.5%) underwent mechanical thrombectomy combined with thrombus aspiration. Based on the literature review and our 10-year experience, we propose an algorithm for the management of intraoperative CA caused by PE. CONCLUSION: The essential components for adequate management of intraoperative PE with CA include hemodynamic support, cardiopulmonary resuscitation, and the implementation of a primary perfusion intervention. The prompt identification of the criteria for each specific treatment modality, guided by the individual patient's characteristics, is necessary for an optimal approach.


Subject(s)
Heart Arrest , Pulmonary Embolism , Thrombosis , Humans , Thrombolytic Therapy/adverse effects , Thrombolytic Therapy/methods , Treatment Outcome , Pulmonary Embolism/complications , Pulmonary Embolism/surgery , Thrombosis/complications , Thrombosis/surgery , Heart Arrest/therapy , Heart Arrest/complications
5.
BMC Neurol ; 21(1): 45, 2021 Jan 30.
Article in English | MEDLINE | ID: mdl-33514324

ABSTRACT

Endovascular procedures with liquid embolic agents such as ethylene-vinyl alcohol (EVOH) copolymers are indicated before surgical treatment of cervical paraganglioma. Consequently, these agents are now available as low viscosity formulations, one of which is Squid 12, which are demonstrating superior vascular penetration. Cases of facial paralysis secondary to embolization of cervical vascular lesions with classic embolic agents have been reported in the English literature, however, this complication has not been described with new generation options such as Squid 12.We describe the case of a 43-year-old patient with a left neck carotid paraganglioma. Embolization was performed under general anaesthesia before surgical excision. In the immediate postoperative period, the patient developed total left facial palsy. Since the imaging tests (Computed Tomography (CT) and Magnetic Resonance Imaging (MRI)) and neurological examination showed no involvement of additional cranial nerves (CN), we hypothesise that the main cause of this complication is ischemia of the vasa nervorum of CN VII secondary to embolization. Almost six months later, the patient continues to present total facial paralysis (Grade VI House-Brackmann facial paralysis scale), and palsy of the left CN X and XII as a complication secondary to surgical resection of the paraganglioma.This case is relevant since it is the first clinical case of permanent facial paralysis secondary to embolization with Squid 12.


Subject(s)
Embolization, Therapeutic/adverse effects , Facial Paralysis/etiology , Head and Neck Neoplasms/therapy , Paraganglioma/therapy , Polyvinyls/adverse effects , Adult , Embolism/chemically induced , Embolization, Therapeutic/methods , Endovascular Procedures/adverse effects , Endovascular Procedures/methods , Female , Humans , Magnetic Resonance Imaging
6.
Rev Esp Enferm Dig ; 113(8): 618, 2021 08.
Article in English | MEDLINE | ID: mdl-33267592

ABSTRACT

The patient had a history of recurrent acute necrotizing pancreatitis, chronic pancreatitis and a pancreatic pseudocyst with an associated pseudoaneurysm in the superior mesenteric artery. He presented mesogastric pain, nausea, 187 U/l amylase and 242 U/l lipase. A hemorrhagic appearance inside the pseudocyst and lumen of the common bile duct was seen on ultrasound.


Subject(s)
Aneurysm, False , Pancreatic Pseudocyst , Pancreatitis, Chronic , Aneurysm, False/complications , Aneurysm, False/diagnostic imaging , Gastrointestinal Hemorrhage/diagnostic imaging , Gastrointestinal Hemorrhage/etiology , Humans , Male , Pancreatic Pseudocyst/complications , Pancreatic Pseudocyst/diagnostic imaging , Pancreatitis, Chronic/complications , Pancreatitis, Chronic/diagnostic imaging
7.
Anal Chem ; 92(20): 13641-13646, 2020 10 20.
Article in English | MEDLINE | ID: mdl-32955253

ABSTRACT

In this letter, the innate ability of nitric oxide (NO) to inhibit platelet activation/adhesion/thrombus formation is employed to improve the hemocompatibility and in vivo accuracy of an intravascular (IV) potentiometric PCO2 (partial pressure of carbon dioxide) sensor. The catheter-type sensor is fabricated by impregnating a segment of dual lumen silicone tubing with a proton ionophore, plasticizer, and lipophilic cation-exchanger. Subsequent filling of bicarbonate and strong buffer solutions and placement of Ag/AgCl reference electrode wires within each lumen, respectively, enables measurement of the membrane potential difference across the inner wall of the tube, with this potential changing as a function of the logarithm of sample PCO2. The dual lumen device is further encapsulated within a S-nitroso-N-acetyl-DL-penicillamine (SNAP)-doped silicone tube that releases physiological levels of NO. The NO releasing sensor exhibits near-Nernstian sensitivity toward PCO2 (slope = 59.31 ± 0.78 mV/decade) and low drift rates (<2 mV/24 h after initial equilibration). In vivo evaluation of the NO releasing sensors, performed in the arteries and veins of anesthetized pigs for 20 h, shows enhanced accuracy (vs non-NO releasing sensors) when benchmarked to measurements of discrete blood samples made with a commercial blood gas analyzer. The accurate, continuous monitoring of blood PCO2 levels achieved with this new IV NO releasing PCO2 sensor configuration could help better manage hospitalized patients in critical care units.


Subject(s)
Biocompatible Materials/chemistry , Carbon Dioxide/analysis , Nitric Oxide/metabolism , Potentiometry/methods , Animals , Blood Vessels/chemistry , Electrodes , Ion Exchange Resins/chemistry , Potentiometry/instrumentation , S-Nitroso-N-Acetylpenicillamine/chemistry , Silicones/chemistry , Swine
8.
Breast Cancer Res Treat ; 180(1): 55-61, 2020 Feb.
Article in English | MEDLINE | ID: mdl-31933142

ABSTRACT

PURPOSE: We quantified cytotoxic T cells in nonmalignant breast tissues from women with and without subsequent breast cancer to assess evidence of whether immunosurveillance may be suppressed prior to tumor development. METHODS: We used an age-matched set of breast tissues from women with benign breast disease (BBD) who subsequently developed breast cancer (BBD with later BC), women with BBD who remained cancer free (BBD cancer-free), and normal Komen Tissue Bank (KTB) tissue donors (KTB controls). We evaluated terminal duct lobular units (lobules) for degree of epithelial abnormality and density of dual-positive CD8/CD103 T cells, as CD103+ cells are thought to be a subset of CD8+ cytotoxic T cells located primarily in the intraepithelial compartment. RESULTS: In 10 sets of age-matched women, 256 breast lobules were studied: 85 in BBD women with later BC, 85 in BBD cancer-free women, and 86 in KTB donors. The majority of all lobules were histologically normal (N = 143, 56%), with 65 (25%) nonproliferative fibrocystic change, and 48 (19%) proliferative epithelial change (with or without atypia). In BBD women with later BC, median CD8+/CD103+ cell density was 39.6, 31.7, and 10.5 cells/mm2 (p = 0.002) for normal, nonproliferative, and proliferative lobules. In BBD cancer-free women, median CD8+/CD103+ cell density values were 46.7, 14.3, and 0 cells/mm2 (p = 0.004) respectively. In KTB donors, CD8+/CD103+ cell density was not significantly different across the lobule types (medians 0, 5.8, 10.7, p = 0.43). CONCLUSION: In women with BBD, breast lobules with increasing epithelial abnormality show significant decreases in cytotoxic T cells as measured by CD8/CD103 staining, suggesting that impaired immunosurveillance may be a component of the earliest stages of breast cancer development.


Subject(s)
Breast Diseases/etiology , Breast Diseases/pathology , Epithelium/metabolism , Epithelium/pathology , T-Lymphocytes, Cytotoxic/immunology , T-Lymphocytes, Cytotoxic/metabolism , Adult , Biomarkers , Breast Neoplasms/etiology , Breast Neoplasms/metabolism , Breast Neoplasms/pathology , Cell Count , Disease Susceptibility/immunology , Female , Follow-Up Studies , Humans , Immunologic Surveillance , Middle Aged , Phenotype
9.
Am J Perinatol ; 36(7): 742-750, 2019 06.
Article in English | MEDLINE | ID: mdl-30372770

ABSTRACT

PURPOSE: We hypothesized that surgical energy could be used to create hysterotomies in open fetal surgery. STUDY DESIGN: Initial studies compared the LigaSure Impact and Harmonic ACE + 7 Shears in the efficiency of hysterotomy and thermal damage. Pregnant ewes at an estimated gestational age (EGA) of 116 to 120 days (term = 145; n = 7) underwent hysterotomy using either device. Hysterotomy edges were resected, and thermal injury extent was determined by histopathological assessment. Upon determining a superior device, subsequent studies compared this to the AutoSuture Premium Poly CS*-57 Stapler in uterine healing. Pregnant ewes (n = 6) at an EGA of 87 to 93 days underwent 6-cm hysterotomy in each gravid horn with either the stapler (n = 5) or Harmonic (n = 5) followed by closure and animal recovery. After 37 to 42 days, uterine healing was assessed by evaluating tensile strength and histopathology. RESULTS: Thermal damage was more extensive with the LigaSure (n = 11 hysterotomies) than with the Harmonic (n = 11; 5.6 ± 1 vs. 3.1 ± 0.6 mm; p < 0.0001);therefore, the Harmonic was selected for healing studies. Gross scar appearance and tensile strength were the same between the Harmonic and stapler. The stapler caused more fibrosis (4/7 samples with "moderate" fibrosis vs. 0/8 with the Harmonic; p = 0.02). CONCLUSION: The Harmonic ACE + 7 caused less thermal injury than the LigaSure Impact and performed similar to the CS*-57 Stapler in uterine healing with continued gestation.


Subject(s)
Electrosurgery/instrumentation , Fetal Therapies/methods , Fetus/surgery , Hysterotomy/methods , Surgical Stapling , Animals , Cicatrix/etiology , Equipment Design , Female , Hysterotomy/adverse effects , Hysterotomy/instrumentation , Models, Animal , Sheep , Uterus/pathology
10.
Breast Cancer Res Treat ; 167(3): 649-658, 2018 02.
Article in English | MEDLINE | ID: mdl-29090365

ABSTRACT

PURPOSE: While the role of natural killer (NK) cells in breast cancer therapy has been investigated, little information is known about NK cell function and presence in nonmalignant and premalignant breast tissue. Here, we investigate and quantify NK cell marker CD56 and activating ligand MICA in breast tissue with benign breast disease. METHODS: Serial tissue sections from 88 subjects, 44 with benign breast disease (BBD) who remained cancer-free, and 44 with BBD who later developed cancer, were stained with H&E, anti-MICA, and anti-CD56. Up to ten representative lobules were identified on each section. Using digital image analysis, MICA and CD56 densities were determined for each lobule, reported as percent of pixels in the lobule that registered as stained by each antibody. Analyses were performed on a per-subject and per-lobule basis. RESULTS: Per-subject multivariate analyses showed associations of CD56 and MICA with age: CD56 was increased in older subjects (p = 0.03), while MICA was increased in younger subjects (p = 0.005). Per-lobule analyses showed that CD56 and MICA levels were both decreased in lobules with fibrocystic change, with median levels of CD56 and MICA staining, respectively, at 0.31 and 7.0% in fibrocystic lobules compared to 0.76 and 12.2% in lobules without fibrocystic change (p < 0.001 for each). Among fibrocystic lobules, proliferative/atypical lobules showed significantly lower expression compared to nonproliferative lobules for MICA (p = 0.02) but not for CD56 (p = 0.80). CONCLUSION: Levels of CD56+ NK cells and activating ligand MICA were decreased in breast lobules with fibrocystic change, and MICA levels showed a significant stepwise decrease with increasing histopathologic abnormality. MICA levels were also significantly decreased in older subjects, who generally have higher risk of developing cancer. These findings advance a model in which MICA promotes cytotoxic activity in CD56+ NK cells to protect against tumorigenesis in breast lobules, and suggest further research is warranted.


Subject(s)
Breast Neoplasms/immunology , Hyperplasia/immunology , Killer Cells, Natural/immunology , Neoplasms/immunology , Adult , Aged , Breast/immunology , Breast/pathology , Breast Neoplasms/genetics , Breast Neoplasms/pathology , CD56 Antigen/immunology , Female , Histocompatibility Antigens Class I/immunology , Humans , Hyperplasia/pathology , Killer Cells, Natural/pathology , Male , Middle Aged , Neoplasms/pathology , Precancerous Conditions/immunology , Precancerous Conditions/pathology
11.
J Environ Manage ; 216: 160-168, 2018 Jun 15.
Article in English | MEDLINE | ID: mdl-28449949

ABSTRACT

Forest residues are an important source of biomass. Among these, Coffee Cut-Stems (CCS) are an abundant wood waste in Colombia obtained from coffee crops renovation. However, only low quantities of these residues are used directly in combustion processes for heating and cooking in coffee farms where their energy efficiency is very low. In the present work, an energy and environmental assessment of two bioenergy production processes (ethanol fermentation and gasification) using CCS as raw material was performed. Biomass gasification seems to be the most promising thermochemical method for bioenergy production whereas, ethanol fermentation is a widely studied biochemical method to produce biofuels. Experimental runs of the CCS gasification were carried out and the synthesis gas composition was monitored. Prior to the fermentation process, a treatment of the CCS is required from which sugar content was determined and then, in the fermentation process, the ethanol yield was calculated. Both processes were simulated in order to obtain the mass and energy balance that are used to assess the energy efficiency and the potential environmental impact (PEI). Moderate high energy efficiency and low environmental impacts were obtained from the CCS gasification. In contrast, high environmental impacts in different categories and low energy efficiencies were calculated from the ethanolic fermentation. Biomass gasification seems to be the most promising technology for the use of Coffee Cut-Stems with high energy yields and low environmental issues.


Subject(s)
Biofuels , Coffee , Biomass , Colombia , Fermentation
12.
Breast Cancer Res Treat ; 164(2): 295-304, 2017 Jul.
Article in English | MEDLINE | ID: mdl-28474262

ABSTRACT

PURPOSE: Atypical ductal hyperplasia (ADH) identified on percutaneous breast biopsy represents a high-risk lesion, upgrading to cancer with surgical excision in ~7-45.8% of cases. Routine excision is questioned due to potential overtreatment and cost. This study evaluates clinical, imaging, and histologic features to predict the risk of upgrade. METHODS: With IRB approval, a single-institution retrospective review was performed of patients who underwent surgical excision of ADH diagnosed by core biopsy from June 2005 to June 2013. We reviewed electronic medical records, breast imaging, and biopsy slides. Multiple imputation was used for missing data. Association of various features with cancer upgrade was assessed using logistic regression. RESULTS: Among 399 cases, the upgrade rate to cancer was 16.0%, (95% CI: 12.8-20.0%), with nine invasive cancers and 55 ductal carcinoma in situ (DCIS) only. Via a logistic regression approach, we defined a subgroup with low risk for upgrade: women whose biopsies showed no individual cell necrosis, and either a) 1 focus of ADH with ≥50% removal, or b) 2-3 foci with ≥90% removal. Cases meeting these criteria had an upgrade rate of 4.9% (95% CI: 1.0-8.9%), compared to 21.4% (16.4-26.3%) in cases that did not meet this low-risk definition. CONCLUSIONS: ADH on core biopsy with low risk of upgrade to cancer is defined by lack of individual cell necrosis, number of foci of ADH, and percent of imaging lesion removed. If these findings are validated, women whose biopsies meet low-risk criteria might be considered for prevention therapy and surveillance without surgical excision.


Subject(s)
Breast Neoplasms/diagnosis , Breast/pathology , Carcinoma, Intraductal, Noninfiltrating/diagnosis , Adult , Aged , Aged, 80 and over , Biopsy, Large-Core Needle , Female , Humans , Logistic Models , Middle Aged , Retrospective Studies , Risk Factors
13.
Mol Pharm ; 14(11): 3762-3771, 2017 11 06.
Article in English | MEDLINE | ID: mdl-29020775

ABSTRACT

A new portable gas phase nitric oxide (NO) generator is described for potential applications in inhaled NO (INO) therapy and during cardiopulmonary bypass (CPB) surgery. In this system, NO is produced at the surface of a large-area mesh working electrode by electrochemical reduction of nitrite ions in the presence of a soluble copper(II)-ligand electron transfer mediator complex. The NO generated is then transported into gas phase by either direct purging with nitrogen/air or via circulating the electrolyte/nitrite solution through a gas extraction silicone fiber-based membrane-dialyzer assembly. Gas phase NO concentrations can be tuned in the range of 5-1000 ppm (parts per million by volume for gaseous species), in proportion to a constant cathodic current applied between the working and counter electrodes. This new NO generation process has the advantages of rapid production times (5 min to steady-state), high Faraday NO production efficiency (ca. 93%), excellent stability, and very low cost when using air as the carrier gas for NO (in the membrane dialyzer configuration), enabling the development of potentially portable INO devices. In this initial work, the new system is examined for the effectiveness of gaseous NO to reduce the systemic inflammatory response (SIR) during CPB, where 500 ppm of NO added to the sweep gas of the oxygenator or to the cardiotomy suction air in a CPB system is shown to prevent activation of white blood cells (granulocytes and monocytes) during extracorporeal circulation with cardiotomy suction conducted with five pigs.


Subject(s)
Cardiopulmonary Bypass/methods , Nitric Oxide/therapeutic use , Administration, Inhalation , Animals , Electrochemistry/methods , Lung/metabolism , Nitrites/chemistry , Swine
15.
EMBO J ; 31(6): 1605-16, 2012 Mar 21.
Article in English | MEDLINE | ID: mdl-22314234

ABSTRACT

The THO complex is a key factor in co-transcriptional formation of export-competent messenger ribonucleoprotein particles, yet its structure and mechanism of chromatin recruitment remain unknown. In yeast, this complex has been described as a heterotetramer (Tho2, Hpr1, Mft1, and Thp2) that interacts with Tex1 and mRNA export factors Sub2 and Yra1 to form the TRanscription EXport (TREX) complex. In this study, we purified yeast THO and found Tex1 to be part of its core. We determined the three-dimensional structures of five-subunit THO complex by electron microscopy and located the positions of Tex1, Hpr1, and Tho2 C-terminus using various labelling techniques. In the case of Tex1, a ß-propeller protein, we have generated an atomic model which docks into the corresponding part of the THO complex envelope. Furthermore, we show that THO directly interacts with nucleic acids through the unfolded C-terminal region of Tho2, whose removal reduces THO recruitment to active chromatin leading to mRNA biogenesis defects. In summary, this study describes the THO architecture, the structural basis for its chromatin targeting, and highlights the importance of unfolded regions of eukaryotic proteins.


Subject(s)
Nucleic Acids/metabolism , Ribonucleoproteins/genetics , Ribonucleoproteins/metabolism , Saccharomyces cerevisiae Proteins/chemistry , Saccharomyces cerevisiae Proteins/metabolism , Transcription Factors/chemistry , Transcription Factors/metabolism , Adenosine Triphosphatases/genetics , Adenosine Triphosphatases/metabolism , Binding Sites , Chromatin/genetics , Chromatin/metabolism , DNA-Binding Proteins/chemistry , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Nuclear Proteins/chemistry , Nuclear Proteins/genetics , Nuclear Proteins/metabolism , Protein Binding , RNA, Messenger/genetics , RNA-Binding Proteins/genetics , RNA-Binding Proteins/metabolism , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , Saccharomyces cerevisiae Proteins/genetics , Transcription Factors/genetics , Transcription, Genetic/genetics
16.
Crit Care Med ; 44(2): e58-69, 2016 Feb.
Article in English | MEDLINE | ID: mdl-26488218

ABSTRACT

OBJECTIVE: To investigate the effects of the combination of extracorporeal cardiopulmonary resuscitation and thrombolytic therapy on the recovery of vital organ function after prolonged cardiac arrest. DESIGN: Laboratory investigation. SETTING: University laboratory. SUBJECTS: Pigs. INTERVENTIONS: Animals underwent 30-minute untreated ventricular fibrillation cardiac arrest followed by extracorporeal cardiopulmonary resuscitation for 6 hours. Animals were allocated into two experimental groups: t-extracorporeal cardiopulmonary resuscitation (t-ECPR) group, which received streptokinase 1 million units, and control extracorporeal cardiopulmonary resuscitation (c-ECPR), which did not receive streptokinase. In both groups, the resuscitation protocol included the following physiologic targets: mean arterial pressure greater than 70 mm Hg, cerebral perfusion pressure greater than 50 mm Hg, PaO2 150 ± 50 torr (20 ± 7 kPa), PaCO2 40 ± 5 torr (5 ± 1 kPa), and core temperature 33°C ± 1°C. Defibrillation was attempted after 30 minutes of extracorporeal cardiopulmonary resuscitation. MEASUREMENTS AND MAIN RESULTS: A cardiac resuscitability score was assessed on the basis of success of defibrillation, return of spontaneous heart beat, weanability from extracorporeal cardiopulmonary resuscitation, and left ventricular systolic function after weaning. The addition of thrombolytic to extracorporeal cardiopulmonary resuscitation significantly improved cardiac resuscitability (3.7 ± 1.6 in t-ECPR vs 1.0 ± 1.5 in c-ECPR). Arterial lactate clearance was higher in t-ECPR than in c-ECPR (40% ± 15% vs 18% ± 21%). At the end of the experiment, the intracranial pressure was significantly higher in c-ECPR than in t-ECPR. Recovery of brain electrical activity, as assessed by quantitative analysis of electroencephalogram signal, and ischemic neuronal injury on histopathologic examination did not differ between groups. Animals in t-ECPR group did not have increased bleeding complications, including intracerebral hemorrhages. CONCLUSIONS: In a porcine model of prolonged cardiac arrest, t-ECPR improved cardiac resuscitability and reduced brain edema, without increasing bleeding complications. However, early electroencephalogram recovery and ischemic neuronal injury were not improved.


Subject(s)
Cardiopulmonary Resuscitation/methods , Extracorporeal Membrane Oxygenation/methods , Fibrinolytic Agents/administration & dosage , Heart Arrest/therapy , Streptokinase/administration & dosage , Animals , Body Temperature , Combined Modality Therapy , Electroencephalography , Fibrinolytic Agents/therapeutic use , Heart Arrest/drug therapy , Hemodynamics , Intracranial Pressure , Streptokinase/therapeutic use , Swine , Time Factors
17.
J Hand Surg Am ; 41(1): 3-12, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26710728

ABSTRACT

PURPOSE: To test the potential for the ex situ limb perfusion system to prolong limb allograft survival up to 24 hours. METHODS: We used 20 swine for the study. In group 1 (control), 4 limbs were perfused with heparin solution and preserved at 4°C for 6 hours. In group 2, 4 limbs were perfused with autologous blood at 27°C to 32°C for 24 hours. In both groups, limbs were transplanted orthotopically to recipients and monitored for 12 hours. In addition to perfusion parameters, we recorded perfusate gases and electrolytes (pH, pCO2, pO2, O2 saturation, Na, K, Cl, Ca, HCO3, glucose, and lactate) and obtained functional electrostimulation hourly throughout the experiment. Histology samples were obtained for TUNEL staining and single-muscle fiber contractility testing. RESULTS: In both groups, hemodynamic variables of circulation remained stable throughout the experiment. Neuromuscular electrical stimulation remained intact until the end of reperfusion in group 2 vs no response in group 1. In group 2, a gradual increase in lactate levels during pump perfusion returned to normal after transplantation. Compared with the contralateral limb in group 2, single-muscle fiber contractility testing showed no significant difference at the end of the experiment. CONCLUSIONS: We demonstrated extended limb survival up to 24 hours using normothermic pulsatile perfusion and autologous blood. CLINICAL RELEVANCE: Successful prolongation of limb survival using ex situ perfusion methods provides with more time for revascularization of an extremity.


Subject(s)
Blood Transfusion, Autologous , Fibrinolytic Agents/administration & dosage , Forelimb/transplantation , Graft Survival , Heparin/administration & dosage , Organ Preservation/methods , Perfusion/methods , Allografts , Amputation, Surgical , Animals , Biopsy , Electric Stimulation , Forelimb/blood supply , Hydrogen-Ion Concentration , Isometric Contraction , Lactic Acid/blood , Models, Animal , Muscle Fibers, Skeletal/pathology , Potassium/blood , Skin Temperature , Swine , Transplantation Conditioning/methods
18.
Anal Chem ; 87(16): 8067-72, 2015 Aug 18.
Article in English | MEDLINE | ID: mdl-26201351

ABSTRACT

A novel electrochemically controlled release method for nitric oxide (NO) (based on electrochemical reduction of nitrite ions) is combined with an amperometric oxygen sensor within a dual lumen catheter configuration for the continuous in vivo sensing of the partial pressure of oxygen (PO2) in blood. The on-demand electrochemical NO generation/release method is shown to be fully compatible with amperometric PO2 sensing. The performance of the sensors is evaluated in rabbit veins and pig arteries for 7 and 21 h, respectively. Overall, the NO releasing sensors measure both venous and arterial PO2 values more accurately with an average deviation of -2 ± 11% and good correlation (R(2) = 0.97) with in vitro blood measurements, whereas the corresponding control sensors without NO release show an average deviation of -31 ± 28% and poor correlation (R(2) = 0.43) at time points >4 h after implantation in veins and >6 h in arteries. The NO releasing sensors induce less thrombus formation on the catheter surface in both veins and arteries (p < 0.05). This electrochemical NO generation/release method could offer a new and attractive means to improve the biocompatibility and performance of implantable chemical sensors.


Subject(s)
Biosensing Techniques/methods , Monitoring, Physiologic/methods , Nitric Oxide/chemistry , Oxygen/analysis , Animals , Electrochemistry/trends , Nitric Oxide/blood , Rabbits , Swine
19.
Plants (Basel) ; 13(2)2024 Jan 19.
Article in English | MEDLINE | ID: mdl-38276757

ABSTRACT

American ginseng (Panax quinquefolius) is widely used due to its medicinal properties. Ontario is a major producer of cultivated American ginseng, where seeds were originally collected from the wild without any subsequent scientific selection, and thus the crop is potentially very diverse. A collection of 162 American ginseng plants was harvested from a small area in a commercial garden and phenotyped for morphological traits, such as root grade, stem length, and fresh and dry weights of roots, leaves, stems, and seeds. All of the traits showed a range of values, and correlations were observed between root and stem weights, root dry weight and leaf dry weight, as well as root and leaf fresh weights. The plants were also genotyped using single nucleotide polymorphisms (SNPs) at the PW16 locus. SNP analysis revealed 22 groups based on sequence relatedness with some groups showing no SNPs and others being more diverse. The SNP groups correlated with significant differences in some traits, such as stem length and leaf weight. This study provides insights into the genetic and phenotypic diversity of cultivated American ginseng grown under similar environmental conditions, and the relationship between different phenotypes, as well as genotype and phenotype, will aid in future selection programs to develop American ginseng cultivars with desirable agronomic traits.

20.
Heliyon ; 10(5): e26515, 2024 Mar 15.
Article in English | MEDLINE | ID: mdl-38434392

ABSTRACT

Rockburst phenomena pose significant challenges in the mining industry, particularly with increased underground activities at greater depths. These sudden failures not only jeopardize personnel safety but also impact mining investments. Consequently, it becomes crucial to assess the reliability and effectiveness of empirical methods employed for predicting rock burst occurrences and their severity, an ongoing subject of debate within the scientific community. This research presents a comprehensive review of empirical approaches for rock burst prediction. Subsequently, these approaches are applied to predict rock burst occurrences and its intensity within sections of a tunnel at the new level of El Teniente mine in Chile. Most of these methods rely on single-factor criteria to predict the likelihood and severity of rock bursts. However, inconsistencies are observed in the results obtained from these approaches in numerous cases. This discrepancy highlights the influence of various input parameters on rock burst estimations and emphasizes that single-index criteria may not encompass all the pertinent factors that contribute to this phenomenon. Consequently, such criteria may inadequately estimate or reflect the probability of rock burst occurrences. Given the multifaceted nature of rock burst phenomena, which depend on multiple factors, it becomes imperative to explore new approaches that consider a broader range of influencing factors, thereby yielding more realistic results. Hence, continued research is essential to develop new methods that address this issue comprehensively and ensure the safety of the mining industry.

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