RESUMEN
BACKGROUND: Children treated for cancer are at risk for adverse effects of iron due to transfusions administered during prolonged marrow suppression, which may increase exposure to toxic forms of iron, extrahepatic iron accumulation, and long-term organ damage. OBJECTIVE: This study aimed to characterize the severity and organ distribution of clinically significant, multisystem iron overload (IO) in an at-risk cohort of pediatric cancer patients. METHODS: This was a retrospective, cross-sectional study of childhood cancer patients who underwent a magnetic resonance imaging (MRI) due to clinical concern for IO. Data regarding cancer type and treatment, transfusion history, MRI and laboratory results, and treatment for IO were collected. Severity of IO was analyzed by non-parametric tests with respect to clinical characteristics. RESULTS: Of the 103 patients, 98% of whom had a Cancer Intensity Treatment Rating (ITR-3) of 3 or higher, 53% (54/102) had moderate or greater hepatic siderosis, 80% (77/96) had pancreatic siderosis, 4% (3/80) had cardiac siderosis, and 45% (13/29) had pituitary siderosis and/or volume loss. Pancreatic iron was associated with both cardiac (p = .0043) and pituitary iron (p = .0101). In the 73 off-therapy patients, ferritin levels were lower (p = .0008) with higher correlation with liver iron concentration (LIC) (p = .0016) than on-therapy patients. Fifty-eight subjects were treated for IO. CONCLUSION: In this heavily treated cohort of pediatric cancer patients, more than 80% had extrahepatic iron loading, which occurs with significant exposure to toxic forms of iron related to decreased marrow activity in setting of transfusions. Further studies should examine the effects of exposure to reactive iron on long-term outcomes and potential strategies for management.
RESUMEN
BACKGROUND: Platelet transfusion is a critical and often necessary aspect of managing cancer. Low platelet counts frequently lead to bleeding complications; however, the drugs used to combat malignancy commonly lead to decreased production and destruction of the very cell whose function is essential to stop bleeding. The transfusion of allogeneic platelet products helps to promote hemostasis, but alloimmunization may make it difficult to manage other complications associated with cancer. METHODS: The literature relating to platelet transfusion in patients with cancer was reviewed. RESULTS: Platelet storage, dosing, transfusion indications, and transfusion response are essential topics for health care professionals to understand because many patients with cancer will require platelet transfusions during the course of treatment. The workup and differentiation of non-immune-mediated compared with immune-mediated platelet refractoriness are vital because platelet management is different between types of refractoriness. CONCLUSIONS: A combination of appropriate utilization of platelet inventory and laboratory testing coupled with communication between those caring for patients with cancer and those providing blood products is essential for effective patient care.
Asunto(s)
Transfusión de Plaquetas/efectos adversos , Transfusión de Plaquetas/métodos , Trombocitopenia/terapia , Plaquetas , Hemorragia/terapia , Humanos , Neoplasias/tratamiento farmacológico , Neoplasias/cirugía , Recuento de PlaquetasRESUMEN
BACKGROUND: In recent years, much attention has been given to the lack of reproducibility in biomedical research, particularly in preclinical animal studies. This is a problem that also plagues the alcohol research field, particularly in consistent consumption in animal models of alcohol use disorders. One often overlooked factor that could affect reproducibility is the maintenance diet used in preclinical studies. METHODS: Herein, 2 well-established models of alcohol consumption, the "drinking in the dark" (DID) procedure and the continuous 2-bottle choice (C2BC) paradigm, were employed to determine the effects of diet on ethanol (EtOH) consumption. Male C57BL/6J mice were given 1 of 6 standard rodent chow diets obtained from Purina LabDiet(®) , Inc. (Prolab(®) RMH 3000) or Harlan(®) Laboratories, Inc. (Teklad Diets T.2916, T.2918, T.2920X, T.7912, or T.8940). A separate group of animals were used to test dietary effects on EtOH pharmacokinetics and behavioral measures following intraperitoneal (IP) injections of various doses of EtOH. RESULTS: Mice eating Harlan diets T.2916 (H2916) and T.2920X (H2920) consumed significantly less EtOH and exhibited lower blood EtOH concentrations (BECs) during DID; however, during C2BC, animals maintained on Harlan T.7912 (H7912) consumed more EtOH and had a higher EtOH preference than the other diet groups. EtOH consumption levels did not stem from changes in alcohol pharmacokinetics, as a separate group of animals administered EtOH IP showed no difference in BECs. However, animals on Harlan diet T.2920X (H2920) were more sensitive to alcohol-induced locomotor activity in an open-field task. No diet-dependent differences were seen in alcohol-induced sedation as measured with loss of righting reflex. CONCLUSIONS: Although these data do not identify a specific mechanism, together, they clearly show that the maintenance diet impacts EtOH consumption. It is incumbent upon the research community to consider the importance of describing nutritional information in methods, which may help decrease interlaboratory reproducibility issues.
Asunto(s)
Consumo de Bebidas Alcohólicas , Alimentación Animal , Consumo Excesivo de Bebidas Alcohólicas , Conducta de Elección , Dieta , Etanol/administración & dosificación , Consumo de Bebidas Alcohólicas/fisiopatología , Alimentación Animal/normas , Animales , Consumo Excesivo de Bebidas Alcohólicas/fisiopatología , Conducta de Elección/efectos de los fármacos , Conducta de Elección/fisiología , Dieta/normas , Ingestión de Alimentos/efectos de los fármacos , Ingestión de Alimentos/fisiología , Masculino , Ratones , Ratones Endogámicos C57BLRESUMEN
Neonatal rodents undergo anesthesia for numerous procedures and for euthanasia by anesthetic overdose. However, data regarding whether neonatal anesthesia is humane are limited. Hypothermia (cryoanesthesia) is the most commonly used anesthetic protocol for neonatal rats 10 d of age or younger. However, hypothermia has recently been restricted in several countries due to perceived painful effects, including pain on rewarming. Minimizing the potential pain and distress of neonates in research is imperative, although very challenging. Traditional validated and nonvalidated behavioral and physiologic outcome measures used for adult rats undergoing anesthesia are unsuitable for evaluating neonates. Therefore, we investigated the effects of several anesthetic methods on neonatal rats by using the innovative objective approaches of noninvasive ultrasonic vocalizations and more invasive neuroendocrine responses (i. e., serum corticosterone, norepinephrine, glucose). Our results show that hypothermia leads to heightened acute distress in neonatal rats as indicated by prolonged recovery times, increased duration of vocalizations, and elevated corticosterone levels, as compared with neonates undergoing inhalational anesthesia. We demonstrate that inhalational anesthesia is preferable to cryoanesthesia for neonatal rats, and researchers using hypothermia anesthesia should consider using inhalational anesthesia as an alternative method.
Asunto(s)
Anestésicos por Inhalación , Hipotermia , Animales , Ratas , Hipotermia/inducido químicamente , Hipotermia/veterinaria , Animales Recién Nacidos , Vocalización Animal , Ultrasonido , Corticosterona , Dolor , Anestesia por Inhalación , Anestésicos por Inhalación/efectos adversosRESUMEN
Blood collection is frequently used for neonatal and juvenile mice in toxicology, developmental, and immunology studies and is often a terminal procedure. However, the use of nonterminal blood collection techniques, including the submandibular and the submental collection techniques described for adult mice, may offer opportunities to reduce animal numbers and refine current methods. The use of the submental technique has not been described for neonatal or juvenile mice. In this study, we compared the submental and submandibular blood collection techniques to determine their suitability for use in neonatal and juvenile mice. Male and female CD1 mice, ages 7, 14, 21, and 28 d, were randomized by sex into submental (n = 16), submandibular (n = 16), or control (n = 8) groups. Each mouse was weighed, bled per its assigned group (or only restrained in the case of control mice), and then decapitated without anesthesia for terminal blood collection. Blood collection volume and corticosterone concentrations were measured. The 2 methods showed significant differences in the volume of blood collected at ages 14 and 28, with the submandibular technique yielding significantly higher volumes. No significant differences were detected in corticosterone levels between the 2 techniques based on age or sex. A subset of mice (n = 8, 2 per age group) were bled via submental or submandibular technique and were evaluated 48 h later for gross and histopathologic evidence of trauma. Seven of the 8 mice showed expected inflammation and healing at the collection sites, with 4 mice having embedded strands of fur in the tissue. These data indicate that the submental blood collection is a viable method for nonterminal blood collection method in neonatal and juvenile mice, especially when smaller amounts of blood are needed.
Asunto(s)
Animales Recién Nacidos , Recolección de Muestras de Sangre , Animales , Ratones , Femenino , Masculino , Recolección de Muestras de Sangre/métodos , Recolección de Muestras de Sangre/veterinaria , Corticosterona/sangre , Distribución Aleatoria , Glándula SubmandibularRESUMEN
PURPOSE: To evaluate the cost-effectiveness of obtaining a preoperative type and screen (T/S) for common urologic procedures. METHODS: A decision tree model was constructed to track surgical patients undergoing two preoperative blood ordering strategies as follows: obtaining a preoperative T/S versus not doing so. The model was applied to the National (Nationwide) Inpatient Sample (NIS) data, from January 1, 2006 to September 30, 2015. Cost estimates for the model were created from combined patient-level data with published costs of a T/S, type and crossmatch (T/C), a unit of pRBC, and one unit of emergency-release transfusion (ERT). The primary outcome was the incremental cost per ERT prevented, expressed as an incremental cost-effectiveness ratio (ICER) between the two preoperative blood ordering strategies. A cost-effectiveness analysis determined the ICER of obtaining preoperative T/S to prevent an emergency-release transfusion (ERT), with a willingness-to-pay threshold of $1,500.00. RESULTS: A total of 4,113,144 surgical admissions from 2006 to 2015 were reviewed. The overall transfusion rate was 10.54% (95% CI, 10.17-10.91) for all procedures. The ICER of preoperative T/S was $1500.00 per ERT prevented. One-way sensitivity analysis demonstrated that the risk of transfusion should exceed 4.12% to justify preoperative T/S. CONCLUSION: Routine preoperative T/S for radical prostatectomy (rate = 3.88%) and penile implants (rate = .91%) does not represent a cost-effective practice for these surgeries. It is important for urologists to review their institution T/S policy to reduce inefficiencies within the preoperative setting.
Asunto(s)
Tipificación y Pruebas Cruzadas Sanguíneas , Transfusión Sanguínea , Masculino , Humanos , Análisis Costo-Beneficio , Transfusión Sanguínea/métodos , Análisis de Costo-Efectividad , Procedimientos Quirúrgicos UrológicosRESUMEN
Germ-free (GF) mice, which are depleted of their resident microbiota, are the gold standard for exploring the role of the microbiome in health and disease; however, they are of limited value in the study of human-specific pathogens because they do not support their replication. Here, we develop GF mice systemically reconstituted with human immune cells and use them to evaluate the role of the resident microbiome in the acquisition, replication and pathogenesis of two human-specific pathogens, Epstein-Barr virus (EBV) and human immunodeficiency virus (HIV). Comparison with conventional (CV) humanized mice showed that resident microbiota enhance the establishment of EBV infection and EBV-induced tumorigenesis and increase mucosal HIV acquisition and replication. HIV RNA levels were higher in plasma and tissues of CV humanized mice compared with GF humanized mice. The frequency of CCR5+ CD4+ T cells throughout the intestine was also higher in CV humanized mice, indicating that resident microbiota govern levels of HIV target cells. Thus, resident microbiota promote the acquisition and pathogenesis of two clinically relevant human-specific pathogens.
RESUMEN
Tissue factor (TF) is crucial for embryogenesis, as mice lacking TF are embryonically lethal (E10.5). This lethality may be attributed to defects in vascular development and circulatory failure, suggesting additional roles for TF in embryonic development beyond coagulation. In this study, we characterized the role of one of the TF paralogs (f3a) using a zebrafish model. The expression of f3a during embryonic developmental stages was determined by RT-PCR. Spatiotemporal expression pattern of f3a revealed (high expression from 28 to 36 hpf) the role of in the development of the yolk sac, circulation, and fins. Morpholinos (MO), an antisense-based oligonucleotide strategy, was used to knockdown f3a and examined for defects in morphological appearance, bleeding, and vascular patterning. f3a MO-injected embryos showed morphological abnormalities, including shorter body lengths and crooked tails. O-dianisidine staining showed f3a MO-injected embryos exhibited bleeding in the trunk (5.44%) and head (9.52%) regions. Imaging of endothelial-specific transgenic lines (flk1:egfp-NLS/kdrl:mCherry-CAAX) showed a 3-fold decreased caudal vein plexus (CVP) in f3a morphants versus controls at 48 hpf, suggesting a potential role for f3a in angiogenesis. These findings confirm that f3a is essential for angiogenesis, in addition to its involvement in hemostasis.
RESUMEN
Recent studies have shown beneficial effects of environmental enrichment (EE) for zebrafish, while infection of zebrafish with the common pathogen Pseudoloma neurophilia has negative effects. This study investigates the effects of P. neurophilia infection and EE in housing and breeding tanks on measures of behavior, growth, and reproduction. Zebrafish were socially housed and were either infected, P. neurophilia-infected (PNI) (n = 12 tanks), or SPF for P. neurophilia (SPF) (n = 24 tanks). Fish were housed with or without EE, which consisted of placing plastic plants in the tanks; sprigs from plants were placed in half of the breeding tanks for half of breedings, alternating breeding tanks without EE weekly. Behavioral testing included the Novel Tank Diving Test (NTT) and Light/Dark Preference Test (LDT) conducted prior to breeding. At the end of the study, biometric data were collected. Histopathology and molecular analysis for common diseases in fish confirmed that SPF fish remained SPF and that fish from all PNI tanks were infected. PNI fish produced significantly fewer eggs and had lower body weights and lengths than did SPF fish. Fish with EE had longer body lengths, than did fish without EE, and male fish had longer body lengths than female fish. The biometric results and reproductive measures show that SPF fish exhibited better growth and suggest that EE in housing tanks could improve fish growth. The behavioral test results were inconclusive regard- ing whether infection status or EE altered anxiety-like behavior. Our results support other recent studies showing negative effects of P. neurophilia infection on zebrafish.
Asunto(s)
Enfermedades de los Peces , Microsporidiosis , Animales , Femenino , Masculino , Microsporidios , Reproducción , Pez CebraRESUMEN
Mice are an invaluable resource for studying virus-induced disease. They are a small, genetically modifiable animal for which a large arsenal of genetic and immunologic tools is available for evaluation of pathogenesis and potential vaccines and therapeutics. SARS-CoV-2, the betacoronavirus responsible for the COVID-19 pandemic, does not naturally replicate in wild-type mice, due to structural differences between human and mouse ACE2, the primary receptor for SARS-CoV-2 entry into cells. However, several mouse strains have been developed that allow for SARS-CoV-2 replication and clinical disease. Two broad strategies have primarily been deployed for developing mouse strains susceptible to COVID-19-like disease: adding in the human ACE2 gene and adapting the virus to the mouse ACE2 receptor. Both approaches result in mice that develop several of the clinical and pathologic hallmarks of COVID-19, including acute respiratory distress syndrome and acute lung injury. In this review, we describe key acute pulmonary and extrapulmonary pathologic changes seen in COVID-19 patients that mouse models of SARS-CoV-2 infection ideally replicate, the essential development of mouse models for the study of Severe Acute Respiratory Syndrome and Middle Eastern Respiratory Syndrome and the basis of many of the models of COVID-19, and key clinical and pathologic features of currently available mouse models of SARS-CoV-2 infection.
Asunto(s)
COVID-19 , Pandemias , Animales , Modelos Animales de Enfermedad , Humanos , Pulmón , Ratones , SARS-CoV-2RESUMEN
Activation of blood coagulation and endothelial inflammation are hallmarks of respiratory infections with RNA viruses that contribute significantly to the morbidity and mortality of patients with severe disease. We investigated how signaling by coagulation proteases affects the quality and extent of the response to the TLR3-ligand poly(I:C) in human endothelial cells. Genome-wide RNA profiling documented additive and synergistic effects of thrombin and poly(I:C) on the expression level of many genes. The most significantly active genes exhibiting synergistic induction by costimulation with thrombin and poly(I:C) included the key mediators of 2 critical biological mechanisms known to promote endothelial thromboinflammatory functions: the initiation of blood coagulation by tissue factor and the control of leukocyte trafficking by the endothelial-leukocyte adhesion receptors E-selectin (gene symbol, SELE) and VCAM1, and the cytokines and chemokines CXCL8, IL-6, CXCL2, and CCL20. Mechanistic studies have indicated that synergistic costimulation with thrombin and poly(I:C) requires proteolytic activation of protease-activated receptor 1 (PAR1) by thrombin and transactivation of PAR2 by the PAR1-tethered ligand. Accordingly, a small-molecule PAR2 inhibitor suppressed poly(I:C)/thrombin-induced leukocyte-endothelial adhesion, cytokine production, and endothelial tissue factor expression. In summary, this study describes a positive feedback mechanism by which thrombin sustains and amplifies the prothrombotic and proinflammatory function of endothelial cells exposed to the viral RNA analogue, poly(I:C) via activation of PAR1/2.
Asunto(s)
Receptor PAR-1 , Trombina , Células Endoteliales , Retroalimentación , Humanos , ARN ViralRESUMEN
Larval, or tadpole-stage Xenopus laevis frogs are a popular research model for developmental biology and disease studies. Existing euthanasia guidance documents offer recommendations for both eggs and adult stages, yet do not specifically address the larval stage. Data evaluating effective euthanasia methods for groups of X. laevis tadpoles would therefore be useful. The goal of the current study was to evaluate the efficacy of various immersion euthanasia procedures on tadpoles: tricaine methanesulfonate (MS222) at 6 g/L, eugenol at 800 µL/L and rapid chilling (2 to 4 °C). We also evaluated tadpoles at various developmental stages (NF stages 46, 47 and 49). Tadpoles (n = 70) were exposed to euthanasia solution for 15 min, and controls (n = 40) were placed in housing tank water for 15 min. All animals were then placed in recovery tanks containing housing tank water for 4 h to confirm irreversibility of each agent. Cessation of the heartbeat was assessed at the end of euthanasia solution exposure and at each hour thereafter. We found that immersion in a 6 g/L solution of MS222 resulted in 100% euthanasia of all larval stages tested. Conversely, eugenol produced variable euthanasia rates that were affected by both age group and batches of stock solutions. Rapid chilling was completely ineffective as a euthanasia method in our study. Based on our findings, we recommend MS222 as an effective and practical means of euthanizing large numbers of X. laevis tadpoles.
Asunto(s)
Aminobenzoatos/administración & dosificación , Eugenol/administración & dosificación , Eutanasia Animal/métodos , Xenopus laevis , Bienestar del Animal , Animales , Frío , Femenino , Guías como Asunto , Larva , Masculino , Xenopus laevis/crecimiento & desarrolloRESUMEN
BACKGROUND: Maternal red cell IgG antibodies can cross the placenta and cause hemolysis of fetal red cells in case of antigenic differences between maternal and fetal RBCs, leading to hemolytic disease of the fetus and newborn (HDFN). Although the incidence of anti-D associated HDFN has drastically reduced with Rh immune globulin prophylaxis, HDFN due to other maternal red cell alloantibodies still remains a concern. Prevalence and specificities of clinically significant red cell alloantibodies in pregnant females have rarely been reported in the USA. METHODS: A retrospective chart review was conducted to determine the prevalence and specificity of clinically significant red cell alloantibodies in pregnant females who delivered at Beaumont Hospital Royal Oak between May 1, 2017 and December 31, 2017. A total of 4548 pregnant females were screened using electronic medical records. One female above 50 years age and two females with invalid ABO type were excluded from the study per IRB approved protocol. The remaining 4545 pregnant females with a valid ABO/RhD type and valid red cell antibody screen were included. RESULTS: Out of the 4545 included females, 440 had a positive red cell antibody screen. Of these 440 females, 34 had clinically significant alloantibodies, giving an overall prevalence of 0.74%. Anti-E was the most frequently identified significant alloantibody followed by anti-K. The most prevalent significant alloantibodies in RhD positive and RhD negative females were anti-E and anti-K, respectively. Significant association (p-value <0.001) was found between RhD type and the presence of clinically significant alloantibodies amongst females with positive antibody screen. CONCLUSION: Our study aims to reiterate the importance of maternal red cell antibody screening during early pregnancy to help identify and manage high-risk pregnancies. Minimizing the exposure of childbearing age females to incompatible red cell antigens through unnecessary transfusions can help reduce the incidence of red cell alloimmunization and the risk of HDFN.
RESUMEN
: The gut microbial metabolite, trimethylamine N-oxide (TMAO), was previously reported to induce platelet hypersensitivity, which leads to thrombotic risk. However, the molecular mechanism underlying the effects of TMAO on endothelial cells (EC), which is the primary vessel wall contact with the lumen, remains unclear. Here, we investigated the impact of TMAO on procoagulant activity (PCA) in EC and mice, for a possible link between microbiota and coagulation. To test the PCA of TMAO in EC, we performed one-stage clotting assays and converted into PCA. Antitissue factor (TF) antibody was used to test the TF role in PCA. Quantitative PCR was performed to measure the TF, thrombomodulin, IL-6, TF pathway inhibitor and IL-1b expressions at mRNA levels. To test the PCA and thrombotic risk by TMAO in mice, we challenged the mice with TMAO (8âmg/kg; 3âh) and measured the thrombin-anti-thrombin complex (TAT) and D-dimer levels as well as ferric chloride (FeCl3)-induced carotid artery thrombosis model. TMAO-induced TF expression in EC at mRNA and protein levels, dose-dependently. TF blocking experiment confirmed that the increased PCA by TMAO is TF-dependent. Also, mitogen-activated protein kinase pathway inhibitors abolished TMAO-induced TF expression. However, TMAO challenged mice failed to develop systemic activation of coagulation (TAT and D-dimer), as well as a FeCl3-induced carotid arterial thrombosis model. Our results indicated that TMAO triggered TF-dependent PCA via activation of nuclear factor-κB and downregulated thrombomodulin expression in human EC, but failed to develop systemic activation of coagulation in mice.
Asunto(s)
Coagulación Sanguínea/efectos de los fármacos , Células Endoteliales/efectos de los fármacos , Hemostasis/efectos de los fármacos , Metilaminas/farmacología , Animales , Células Cultivadas , Humanos , Ratones , FN-kappa B/metabolismo , Oxidantes/farmacología , Trombomodulina/efectos de los fármacos , Trombomodulina/metabolismoRESUMEN
BACKGROUND Platelet transfusion is a common clinical practice required for therapeutic purposes in the setting of symptomatic thrombocytopenia, and, in some cases, prophylactically for asymptomatic thrombocytopenia. Crossmatch compatibility is not routinely done for platelet transfusions, and transfusion of ABO non-identical platelets has been adapted as an acceptable clinical practice. Acute intravascular hemolysis due to ABO non-identical platelets is a rare but clinically significant entity. Our case report reinforces the importance of a vigilant clinical approach in case of ABO non-identical platelet transfusions. CASE REPORT We report the case of 61-year-old woman with blood group A, with chemotherapy-induced asymptomatic thrombocytopenia, who developed acute intravascular hemolysis following transfusion of group O single-donor platelets (SDPs). The patient was transfused 1 unit of single-donor platelets for bleeding prophylaxis, as her platelet count dropped to less than 10×109/L due to chemotherapy that she was receiving for acute myeloid leukemia (AML). Immediately after transfusion, the patient noticed cherry-colored urine; and within 12 h of transfusion, her hemoglobin dropped by more than 2.5 g/dL. A post-transfusion immunohematology work-up showed positive DAT and high titers of anti-A1 isohemagglutinins in the platelet donor, supporting the diagnosis of acute intravascular hemolysis due to ABO non-identical platelets. CONCLUSIONS The possibility of acute intravascular hemolysis should be kept in mind in cases of transfusion of group O single donor platelets to non-group O recipients. ABO non-identical platelets, even with low isohemagglutinin titers, can cause significant adverse effects, particularly in newborns, children, and immunosuppressed and transfusion-dependent patients; therefore, a cautious clinical approach is recommended.
Asunto(s)
Sistema del Grupo Sanguíneo ABO , Incompatibilidad de Grupos Sanguíneos , Hemólisis , Transfusión de Plaquetas , Femenino , Humanos , Huésped Inmunocomprometido , Persona de Mediana Edad , Trombocitopenia/terapiaRESUMEN
Floor contamination control practices in rodent housing facilities commonly include disposable shoe covers despite the lack of evidence for their usefulness in bioexclusion. Contamination control flooring mats are advertised as an economical and environmentally-responsible alternative to shoe covers, yet little is published regarding their efficacy in preventing the transfer of organic material and the introduction of infectious agents into facilities. We evaluated 4 floor contamination control strategies-shoe covers (ShCv), contamination control flooring (CCF), using both products concurrently (ShCv+CCF) compared with using neither-in preventing bacterial transfer and reducing organic load on facility floors and maintaining murine colony health status. According to PCR assay and culture analysis, ShCv provided the greatest reduction in bacte- rial numbers. Either ShCv, CCF, or ShCv+CCF significantly decreased ATP levels within the facility compared with those at facility entrances, with ShCv+CCF yielding the greatest reduction; however, even when neither ShCv nor CCF was used, intrafacility floor ATP levels were about half those at entrances. According to PCR analyses, no murine parasitic, viral, and bacterial pathogens excluded at the institution were detected in any floor, exhaust air dust, or sentinel samples at any time or location, regardless of the floor contamination control method in use. These findings show that floor contamination control methods help to reduce the organic load in rodent IVC facilities but do not enhance protection from environmental contamination due to murine pathogens.
Asunto(s)
Crianza de Animales Domésticos , Microbiología Ambiental , Pisos y Cubiertas de Piso , Ciencia de los Animales de Laboratorio , Ropa de Protección , Enfermedades de los Roedores/prevención & control , Animales , Monitoreo del Ambiente , Humanos , RatonesAsunto(s)
Sistema del Grupo Sanguíneo ABO/análisis , Tipificación y Pruebas Cruzadas Sanguíneas/instrumentación , Tipificación y Pruebas Cruzadas Sanguíneas/métodos , Computadores , Sistema del Grupo Sanguíneo ABO/sangre , Incompatibilidad de Grupos Sanguíneos/sangre , Incompatibilidad de Grupos Sanguíneos/diagnóstico , Incompatibilidad de Grupos Sanguíneos/prevención & control , Tipificación y Pruebas Cruzadas Sanguíneas/normas , Técnicas de Laboratorio Clínico/instrumentación , Técnicas de Laboratorio Clínico/métodos , Toma de Decisiones Asistida por Computador , Humanos , Práctica Profesional/normas , Estados Unidos , United States Food and Drug Administration/legislación & jurisprudencia , United States Food and Drug Administration/normas , Estudios de Validación como AsuntoRESUMEN
Endothelial exocytosis of granules is a rapid response to vascular injury. However, the molecular machinery that regulates exocytosis in endothelial cells is not well understood. Recently developed techniques have defined the endothelial proteins that control vesicle and granule trafficking in endothelial cells. These techniques have revealed that syntaxin 4, synaptobrevin 3, and N-ethylmaleimide-sensitive factor (NSF) play a critical role in endothelial granule exocytosis. Additional studies have shown that nitric oxide regulates exocytosis by chemically modifying NSF. Further characterization of the factors that regulate exocytosis will lead to novel treatments for vascular diseases such as myocardial infarction and stroke.
Asunto(s)
Bioensayo/métodos , Células Endoteliales/metabolismo , Exocitosis , Proteínas Sensibles a N-Etilmaleimida/metabolismo , Vesículas Secretoras/metabolismo , Animales , Adhesión Celular , Separación Celular , Células Cultivadas , Células Endoteliales/química , Ensayo de Inmunoadsorción Enzimática , Citometría de Flujo , Células HL-60 , Humanos , Inmunoprecipitación , Rodamiento de Leucocito , Leucocitos/metabolismo , Ratones , Microscopía por Video , Proteínas Sensibles a N-Etilmaleimida/genética , Proteínas Sensibles a N-Etilmaleimida/aislamiento & purificación , Óxido Nítrico/metabolismo , Fragmentos de Péptidos/metabolismo , Proteínas Qa-SNARE/metabolismo , Proteínas Recombinantes de Fusión/metabolismo , Factores de Tiempo , Proteína 3 de Membrana Asociada a Vesículas/metabolismo , Productos del Gen tat del Virus de la Inmunodeficiencia Humana/metabolismoRESUMEN
Association between elevated levels of systemic trimethylamine N-oxide (TMAO) and increased risk for adverse cardiovascular events have been proposed in recent years. Increasing experimental and clinical evidence in the last decade has implicated TMAO as an important contributor to the pathogenesis of cardiovascular diseases. TMAO, the oxygenated product of trimethylamine (TMA), belongs to the class of amine oxides. Most of the TMA derived from the metabolism of choline and L-carnitine by gut bacteria is absorbed into the bloodstream and gets rapidly oxidized to TMAO by the hepatic enzyme, flavin-containing monooxgenase-3. Here, we discussed the biosynthesis of TMAO and clinical studies that have assessed TMAO as a biomarker for various cardiovascular and other diseases such as kidney failure, thrombosis, atherosclerosis, obesity, diabetes and cancer. We also summarized the interaction of TMAO with synthetic and traditional molecules that together affect circulating TMAO levels. LINKED ARTICLES: This article is part of a themed section on Spotlight on Small Molecules in Cardiovascular Diseases. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v175.8/issuetoc.
Asunto(s)
Metilaminas/metabolismo , Animales , Encefalopatías/metabolismo , Insuficiencia Cardíaca/metabolismo , Humanos , Neoplasias/metabolismo , Obesidad/metabolismo , Trombosis/metabolismoRESUMEN
Aggression among mice remains a common undesirable problem in laboratory settings, and animal welfare and scientific outcomes may become compromised depending on the severity of aggression. This study evaluated the effect of cage enrichment comprising a bilevel, mounted 'mezzanine' compared with a cotton square or shelter on intracage male aggression over a 6-wk period. Our first study involved home-cage behavioral challenges to male mice from a high aggression substrain (BALB/cJ) and low-aggression substrain (BALB/cByJ). Aggressive interactions and locomotor activity were scored manually and then compared with measures of activity obtained by using a continuous automated home-cagemonitoring system, the Digital Ventilated Caging (DVC) system. BALB/cJ mice exhibited similar levels of aggression acrosshousing conditions, whereas BALB/cByJ mice had lower aggression when housed with a mezzanine. In the second study,videorecordings and continuous DVC automated measures were collected over 24 h and divided into 12-h light and dark phases. BALB/cByJ mice-but not BALB/cJ-mice had increased aggressive behaviors during the dark phase. However, the DVC detected higher activity levels during the dark phase, compared with the light phase, in both substrains. Elevated activity levels recorded by the DVC correlated with fighting bouts and high levels of locomotion. These results show that a bilevel structural form of enrichment reduces aggression, depending on the BALB/c substrain, and confirms higher aggression levels in the BALB/cJ substrain. In addition, our findings provide evidence that the DVC is effective in identifying mouse cages with patterns of high activity levels, signaling possible aggression incidences, thus potentially allowing for early intervention and consequently improving animal welfare.