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1.
Cell ; 174(3): 730-743.e22, 2018 07 26.
Artículo en Inglés | MEDLINE | ID: mdl-30033368

RESUMEN

Drosophila melanogaster has a rich repertoire of innate and learned behaviors. Its 100,000-neuron brain is a large but tractable target for comprehensive neural circuit mapping. Only electron microscopy (EM) enables complete, unbiased mapping of synaptic connectivity; however, the fly brain is too large for conventional EM. We developed a custom high-throughput EM platform and imaged the entire brain of an adult female fly at synaptic resolution. To validate the dataset, we traced brain-spanning circuitry involving the mushroom body (MB), which has been extensively studied for its role in learning. All inputs to Kenyon cells (KCs), the intrinsic neurons of the MB, were mapped, revealing a previously unknown cell type, postsynaptic partners of KC dendrites, and unexpected clustering of olfactory projection neurons. These reconstructions show that this freely available EM volume supports mapping of brain-spanning circuits, which will significantly accelerate Drosophila neuroscience. VIDEO ABSTRACT.


Asunto(s)
Mapeo Encefálico/métodos , Conectoma/métodos , Red Nerviosa/anatomía & histología , Animales , Encéfalo/anatomía & histología , Encéfalo/diagnóstico por imagen , Dendritas , Drosophila melanogaster/anatomía & histología , Femenino , Microscopía Electrónica/métodos , Cuerpos Pedunculados , Neuronas , Olfato/fisiología , Programas Informáticos
2.
Nature ; 593(7859): 379-384, 2021 05.
Artículo en Inglés | MEDLINE | ID: mdl-34012086

RESUMEN

Chemical reactions represent a class of quantum problems that challenge both the current theoretical understanding and computational capabilities1. Reactions that occur at ultralow temperatures provide an ideal testing ground for quantum chemistry and scattering theories, because they can be experimentally studied with unprecedented control2, yet display dynamics that are highly complex3. Here we report the full product state distribution for the reaction 2KRb → K2 + Rb2. Ultracold preparation of the reactants allows us complete control over their initial quantum degrees of freedom, whereas state-resolved, coincident detection of both products enables the probability of scattering into each of the 57 allowed rotational state-pairs to be measured. Our results show an overall agreement with a state-counting model based on statistical theory4-6, but also reveal several deviating state-pairs. In particular, we observe a strong suppression of population in the state-pair closest to the exoergicity limit as a result of the long-range potential inhibiting the escape of products. The completeness of our measurements provides a benchmark for quantum dynamics calculations beyond the current state of the art.

3.
Clin Infect Dis ; 78(Suppl 1): S71-S75, 2024 01 31.
Artículo en Inglés | MEDLINE | ID: mdl-38294113

RESUMEN

Tularemia is a disease caused by Francisella tularensis, a highly infectious bacteria that can be transmitted to humans by direct contact with infected animals. Because of the potential for zoonotic transmission of F. tularensis, veterinary occupational risk is a concern. Here, we report on a human case of tularemia in a veterinarian after an accidental needlestick injury during abscess drainage in a sick dog. The veterinarian developed ulceroglandular tularemia requiring hospitalization but fully recovered after abscess drainage and a course of effective antibiotics. To systematically assess veterinary occupational transmission risk of F. tularensis, we conducted a survey of veterinary clinical staff after occupational exposure to animals with confirmed tularemia. We defined a high-risk exposure as direct contact to the infected animal's body fluids or potential aerosol inhalation without use of standard personal protective equipment (PPE). Survey data included information on 20 veterinary occupational exposures to animals with F. tularensis in 4 states. Veterinarians were the clinical staff most often exposed (40%), followed by veterinarian technicians and assistants (30% and 20%, respectively). Exposures to infected cats were most common (80%). Standard PPE was not used during 80% of exposures; a total of 7 exposures were categorized as high risk. Transmission of F. tularensis in the veterinary clinical setting is possible but overall risk is likely low. Veterinary clinical staff should use standard PPE and employ environmental precautions when handling sick animals to minimize risk of tularemia and other zoonotic infections; postexposure prophylaxis should be considered after high-risk exposures to animals with suspected or confirmed F. tularensis infection to prevent tularemia.


Asunto(s)
Francisella tularensis , Exposición Profesional , Tularemia , Humanos , Animales , Perros , Tularemia/microbiología , Tularemia/veterinaria , Absceso , Zoonosis/microbiología
4.
Clin Chem Lab Med ; 61(10): 1820-1828, 2023 09 26.
Artículo en Inglés | MEDLINE | ID: mdl-37036784

RESUMEN

OBJECTIVES: Monitoring estradiol (E2) is important for determining the onset of pubertal development as well as in the evaluation of girls with precocious puberty. However, E2 measurement remains an analytical challenge in children, who have lower circulating levels. We developed and evaluated a simple and sensitive LC-MS/MS procedure for serum E2 quantification in pediatric populations and established age- and sex-specific pediatric reference intervals. METHODS: Residual patient serum samples were used to evaluate the analytical performance of our in-house LC-MS/MS E2 assay. The evaluation included accuracy, precision, linearity, functional sensitivity (LLoQ), and method comparison. Age- and sex-specific pediatric E2 reference intervals were also established from a cohort of 405 healthy children (birth to 18 years) recruited with informed consent. Age- and sex-specific differences were assessed, and outliers were removed. Reference intervals were established using the robust method. RESULTS: The assay imprecision was <5.3 %. Assay linearity ranged from 13.7 to 1923.3 pmol/L. The LLoQ corresponding to a CV of 20 % was determined to be 8.9 pmol/L. Bland-Altman analysis revealed a mean bias of 29.3 pmol/L or 9.1 % between our LC-MS/MS E2 assay and an external reference laboratory measuring E2 by LC-MS/MS. CONCLUSIONS: Our LC-MS/MS E2 assay shows acceptable accuracy, precision, functional sensitivity (LLoQ), and linearity for E2 quantification. Our LC-MS/MS E2 assay also showed good agreement with an external reference laboratory measuring E2 by LC-MS/MS. In addition, using CALIPER samples, we established robust age- and sex-specific pediatric E2 reference intervals to improve accuracy of test result interpretation and clinical decision making.


Asunto(s)
Estrona , Espectrometría de Masas en Tándem , Masculino , Femenino , Humanos , Niño , Adolescente , Espectrometría de Masas en Tándem/métodos , Cromatografía Liquida/métodos , Estradiol
5.
Respir Res ; 23(1): 8, 2022 Jan 12.
Artículo en Inglés | MEDLINE | ID: mdl-35022042

RESUMEN

BACKGROUND: Excess pulmonary iron has been implicated in the pathogenesis of lung disease, including asthma and COPD. An association between higher iron content in sputum macrophages and infective exacerbations of COPD has previously been demonstrated. OBJECTIVES: To assess the mechanisms of pulmonary macrophage iron sequestration, test the effect of macrophage iron-loading on cellular immune function, and prospectively determine if sputum hemosiderin index can predict infectious exacerbations of COPD. METHODS: Intra- and extracellular iron was measured in cell-line-derived and in freshly isolated sputum macrophages under various experimental conditions including treatment with exogenous IL-6 and hepcidin. Bacterial uptake and killing were compared in the presence or absence of iron-loading. A prospective cohort of COPD patients with defined sputum hemosiderin indices were monitored to determine the annual rate of severe infectious exacerbations. RESULTS: Gene expression studies suggest that airway macrophages have the requisite apparatus of the hepcidin-ferroportin axis. IL-6 and hepcidin play roles in pulmonary iron sequestration, though IL-6 appears to exert its effect via a hepcidin-independent mechanism. Iron-loaded macrophages had reduced uptake of COPD-relevant organisms and were associated with higher growth rates. Infectious exacerbations were predicted by sputum hemosiderin index (ß = 0.035, p = 0.035). CONCLUSIONS: We demonstrate in-vitro and population-level evidence that excess iron in pulmonary macrophages may contribute to recurrent airway infection in COPD. Specifically, IL-6-dependent iron sequestration by sputum macrophages may result in immune cell dysfunction and ultimately lead to increased frequency of infective exacerbation.


Asunto(s)
Hierro/metabolismo , Macrófagos Alveolares/metabolismo , Enfermedad Pulmonar Obstructiva Crónica/metabolismo , Esputo/metabolismo , Anciano , Femenino , Estudios de Seguimiento , Humanos , Recuento de Leucocitos , Macrófagos Alveolares/patología , Masculino , Estudios Prospectivos , Enfermedad Pulmonar Obstructiva Crónica/patología , Recurrencia
6.
South Med J ; 114(10): 630-635, 2021 10.
Artículo en Inglés | MEDLINE | ID: mdl-34599340

RESUMEN

OBJECTIVES: The objective of this analysis was to gauge how the incidence and mortality of uterine cancer in Kentucky have changed from 1995 through 2017. An assessment of the trends in incidence and mortality across different geographic areas and between different races was also performed. METHODS: Age-adjusted annual incidence and mortality rates for uterine cancer were obtained from the Kentucky Cancer Registry. A meta-regression framework was used to assess changes in incidence and mortality rates during the time frame and to determine differences in these rates between rural versus urban counties, Appalachian versus non-Appalachian counties, and Black versus White women. RESULTS: The incidence of uterine cancer has significantly increased throughout the state of Kentucky since 1995. Uterine cancer incidence was 10% and 22% higher in rural and Appalachian counties, respectively, compared with urban and non-Appalachian counties (P < 0.0001) from 1995 through 2017. In contrast, urban and non-Appalachian women had higher or equivalent age-adjusted mortality from uterine cancer, compared with rural and Appalachian women, respectively. The incidence of uterine cancer was significantly higher in White women compared with Black women from 1995 through 2006, but since 2007, there has been no significant difference in uterine cancer incidence based on race. Black women had higher age-adjusted mortality than White women throughout the entire time period examined. CONCLUSIONS: The incidence of uterine cancer is higher in rural and Appalachian Kentucky, without a corresponding geographic trend in mortality. Uterine cancer mortality is significantly higher in Black women.


Asunto(s)
Mortalidad/tendencias , Neoplasias Uterinas/diagnóstico , Neoplasias Uterinas/mortalidad , Adulto , Femenino , Humanos , Incidencia , Kentucky/epidemiología , Persona de Mediana Edad , Sistema de Registros/estadística & datos numéricos , Neoplasias Uterinas/epidemiología
7.
Gynecol Oncol ; 158(2): 446-451, 2020 08.
Artículo en Inglés | MEDLINE | ID: mdl-32456989

RESUMEN

OBJECTIVES: The goal of this study was to assess how the incidence and mortality of cervical cancer in Kentucky has changed from 1995 through 2017. Additionally, trends in incidence and mortality across different geographic areas and between different races were evaluated. METHODS: Age-adjusted annual incidence and mortality rates for cervical cancer were collected from the Kentucky Cancer Registry (KCR). A quadratic fit model was used to evaluate changes in the incidence and mortality over time and to compare differences in cervical cancer incidence and mortality by: 1) rural versus urban counties, 2) Appalachian versus non-Appalachian counties, and 3) black versus white women. RESULTS: Overall, the incidence of cervical cancer has significantly decreased throughout Kentucky since 1995. When comparing different geographic regions, the incidence was 14% and 23% higher in rural and Appalachian counties, respectively, compared to urban and non-Appalachian counties (p < 0.0001) throughout the study period. The incidence of cervical cancer was significantly higher in black women compared to white women from 1995 through 2007, but since 2008 there has been no significant difference in cervical cancer incidence based on race. Similar to incidence rates, mortality from cervical cancer was 29% higher in Appalachia (p = 0.0004) throughout the studied time period. Black women had higher age-adjusted mortality than white women until 2012, but since that time there has not been a significant difference in cervical cancer mortality based on race. CONCLUSIONS: Women residing in rural and Appalachian Kentucky have higher cervical cancer incidence and mortality rates.


Asunto(s)
Neoplasias del Cuello Uterino/epidemiología , Neoplasias del Cuello Uterino/mortalidad , Adulto , Anciano , Región de los Apalaches/epidemiología , Población Negra/estadística & datos numéricos , Femenino , Humanos , Incidencia , Kentucky/epidemiología , Persona de Mediana Edad , Modelos Estadísticos , Áreas de Pobreza , Sistema de Registros , Población Blanca/estadística & datos numéricos
8.
J Bioenerg Biomembr ; 49(1): 113-119, 2017 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-27665468

RESUMEN

Calcium (Ca2+) plays diverse roles in all living organisms ranging from bacteria to humans. It is a structural element for bones, an essential mediator of excitation-contraction coupling, and a universal second messenger in the regulation of ion channel, enzyme and gene expression activities. In mitochondria, Ca2+ is crucial for the control of energy production and cellular responses to metabolic stress. Ca2+ uptake by the mitochondria occurs by the uniporter mechanism. The Mitochondrial Ca2+ Uniporter (MCU) protein has recently been identified as a core component responsible for mitochondrial Ca2+ uptake. MCU knockout (MCU KO) studies have identified a number of important roles played by this high capacity uptake pathway. Interestingly, this work has also shown that MCU-mediated Ca2+ uptake is not essential for vital cell functions such as muscle contraction, energy metabolism and neurotransmission. Although mitochondrial Ca2+ uptake was markedly reduced, MCU KO mitochondria still contained low but detectable levels of Ca2+. In view of the fundamental importance of Ca2+ for basic cell signalling, this finding suggests the existence of other currently unrecognized pathways for Ca2+ entry. We review the experimental evidence for the existence of alternative Ca2+ influx mechanisms and propose how these mechanisms may play an integral role in mitochondrial Ca2+ signalling.


Asunto(s)
Calcio/metabolismo , Transporte Iónico , Mitocondrias/metabolismo , Calcio/fisiología , Redes y Vías Metabólicas , Dilatación Mitocondrial
9.
J Neurosci Res ; 94(11): 1076-83, 2016 11.
Artículo en Inglés | MEDLINE | ID: mdl-27638593

RESUMEN

Newborn screening (NBS) for Krabbe's disease (KD) has been instituted in several states, and New York State has had the longest experience. After an initial screening of dried blood spots, samples from individuals with galactocerebrosidase (GALC) values below a given cutoff level were subjected to additional testing, including sequencing of the GALC gene. This resulted in the identification of mutations that had previously been found in confirmed KD patients and of variants that had never previously been reported. Some individuals had variants considered to be polymorphisms, alone or on the same allele as another mutation. To help with counseling of families on the risk for a newborn to develop KD, expression studies were conducted with these variants identified by NBS. GALC activity was measured in COS1 cells for 140 constructs and compared with mutations that had previously been seen in confirmed cases of KD. When a polymorphism was present on the same allele as the variant, expressed activity was measured with and without the polymorphism. In some cases the presence of the polymorphism greatly lowered the measured GALC activity, possibly making it disease causing. Although it is not possible to predict conclusively whether a variant is severe and will result in infantile KD if two such variants are present or whether a variant is mild and will result in late-onset disease, some variants clearly are not disease causing. This is the largest expression study of GALC variants/mutations found in NBS and confirmed KD cases. This work will be helpful for counseling families of screen-positive newborns found to have low GALC activity. © 2016 Wiley Periodicals, Inc.


Asunto(s)
Galactosilceramidasa/genética , Leucodistrofia de Células Globoides/genética , Mutación/genética , Animales , Células COS , Cercopithecus , Femenino , Galactosilceramidasa/metabolismo , Pruebas Genéticas , Haplotipos , Humanos , Recién Nacido , Masculino , Mutagénesis Sitio-Dirigida/métodos , New York , Transfección
10.
Genet Med ; 18(3): 239-48, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26795590

RESUMEN

PURPOSE: Krabbe disease (KD) results from galactocerebrosidase (GALC) deficiency. Infantile KD symptoms include irritability, progressive stiffness, developmental delay, and death. The only potential treatment is hematopoietic stem cell transplantation. New York State (NYS) implemented newborn screening for KD in 2006. METHODS: Dried blood spots from newborns were assayed for GALC enzyme activity using mass spectrometry, followed by molecular analysis for those with low activity (≤12% of the daily mean). Infants with low enzyme activity and one or more mutations were referred for follow-up diagnostic testing and neurological examination. RESULTS: Of >1.9 million screened, 620 infants were subjected to molecular analysis and 348 were referred for diagnostic testing. Five had enzyme activities and mutations consistent with infantile KD and manifested clinical/neurodiagnostic abnormalities. Four underwent transplantation, two are surviving with moderate to severe handicaps, and two died from transplant-related complications. The significance of many sequence variants identified is unknown. Forty-six asymptomatic infants were found to be at moderate to high risk for disease. CONCLUSIONS: The positive predictive value of KD screening in NYS is 1.4% (5/346) considering confirmed infantile cases. The incidence of infantile KD in NYS is approximately 1 in 394,000, but it may be higher for later-onset forms.


Asunto(s)
Galactosilceramidasa/genética , Galactosilceramidasa/metabolismo , Leucodistrofia de Células Globoides/diagnóstico , Tamizaje Neonatal/métodos , Polimorfismo de Nucleótido Simple , Algoritmos , Pruebas con Sangre Seca , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Humanos , Recién Nacido , Leucodistrofia de Células Globoides/enzimología , Leucodistrofia de Células Globoides/terapia , Espectrometría de Masas , New York , Valor Predictivo de las Pruebas , Resultado del Tratamiento
11.
Phys Rev Lett ; 116(23): 235301, 2016 Jun 10.
Artículo en Inglés | MEDLINE | ID: mdl-27341242

RESUMEN

We report on the site-resolved observation of characteristic states of the two-dimensional repulsive Fermi-Hubbard model, using ultracold ^{40}K atoms in an optical lattice. By varying the tunneling, interaction strength, and external confinement, we realize metallic, Mott-insulating, and band-insulating states. We directly measure the local moment, which quantifies the degree of on-site magnetization, as a function of temperature and chemical potential. Entropies per particle as low as 0.99(6)k_{B} indicate that nearest-neighbor antiferromagnetic correlations should be detectable using spin-sensitive imaging.

12.
Proc Natl Acad Sci U S A ; 110(48): 19194-201, 2013 Nov 26.
Artículo en Inglés | MEDLINE | ID: mdl-24255111

RESUMEN

We present in vivo images of the human brain acquired with an ultralow field MRI (ULFMRI) system operating at a magnetic field B0 ~ 130 µT. The system features prepolarization of the proton spins at Bp ~ 80 mT and detection of the NMR signals with a superconducting, second-derivative gradiometer inductively coupled to a superconducting quantum interference device (SQUID). We report measurements of the longitudinal relaxation time T1 of brain tissue, blood, and scalp fat at B0 and Bp, and cerebrospinal fluid at B0. We use these T1 values to construct inversion recovery sequences that we combine with Carr-Purcell-Meiboom-Gill echo trains to obtain images in which one species can be nulled and another species emphasized. In particular, we show an image in which only blood is visible. Such techniques greatly enhance the already high intrinsic T1 contrast obtainable at ULF. We further present 2D images of T1 and the transverse relaxation time T2 of the brain and show that, as expected at ULF, they exhibit similar contrast. Applications of brain ULFMRI include integration with systems for magnetoencephalography. More generally, these techniques may be applicable, for example, to the imaging of tumors without the need for a contrast agent and to modalities recently demonstrated with T1ρ contrast imaging (T1 in the rotating frame) at fields of 1.5 T and above.


Asunto(s)
Encéfalo/anatomía & histología , Encéfalo/fisiología , Imagen por Resonancia Magnética/métodos , Magnetometría/instrumentación , Humanos , Modelos Teóricos , Factores de Tiempo
13.
Phys Rev Lett ; 114(19): 193001, 2015 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-26024169

RESUMEN

We realize a quantum-gas microscope for fermionic ^{40}K atoms trapped in an optical lattice, which allows one to probe strongly correlated fermions at the single-atom level. We combine 3D Raman sideband cooling with high-resolution optics to simultaneously cool and image individual atoms with single-lattice-site resolution at a detection fidelity above 95%. The imaging process leaves the atoms predominantly in the 3D motional ground state of their respective lattice sites, inviting the implementation of a Maxwell's demon to assemble low-entropy many-body states. Single-site-resolved imaging of fermions enables the direct observation of magnetic order, time-resolved measurements of the spread of particle correlations, and the detection of many-fermion entanglement.

14.
BMC Public Health ; 15: 567, 2015 Jun 19.
Artículo en Inglés | MEDLINE | ID: mdl-26088616

RESUMEN

BACKGROUND: With health care shifting away from the traditional sick care model, many hospitals are integrating fitness facilities and programs into their clinical services in order to support health promotion and disease prevention at the community level. Through a series of focus groups, the present study assessed characteristics of hospital-affiliated integrated facilities located in Northeast Ohio, United States and members' experiences with respect to these facilities. METHODS: Adult members were invited to participate in a focus group using a recruitment flyer. A total of 6 focus groups were conducted in 2013, each lasting one hour, ranging from 5 to 12 participants per group. The responses and discussions were recorded and transcribed verbatim, then analyzed independently by research team members. Major themes were identified after consensus was reached. RESULTS: The participants' average age was 57, with 56.8% currently under a doctor's care. Four major themes associated with integrated facilities and members' experiences emerged across the six focus groups: 1) facility/program, 2) social atmosphere, 3) provider, and 4) member. Within each theme, several sub-themes were also identified. A key feature of integrated facilities is the availability of clinical and fitness services "under one roof". Many participants remarked that they initially attended physical therapy, becoming members of the fitness facility afterwards, or vice versa. The participants had favorable views of and experiences with the superior physical environment and atmosphere, personal attention, tailored programs, and knowledgeable, friendly, and attentive staff. In particular, participants favored the emphasis on preventive care and the promotion of holistic health and wellness. CONCLUSIONS: These results support the integration of wellness promotion and programming with traditional medical care and call for the further evaluation of such a model with regard to participants' health outcomes.


Asunto(s)
Prestación Integrada de Atención de Salud , Promoción de la Salud , Servicio Ambulatorio en Hospital , Satisfacción del Paciente , Anciano , Femenino , Grupos Focales , Humanos , Masculino , Persona de Mediana Edad , Ohio , Investigación Cualitativa , Estados Unidos
15.
Ann Emerg Med ; 63(1): 1-5, 2014 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-24094476

RESUMEN

STUDY OBJECTIVE: The CricKey is a novel surgical cricothyroidotomy device combining the functions of a tracheal hook, stylet, dilator, and bougie incorporated with a Melker airway cannula. This study compares surgical cricothyroidotomy with standard open surgical versus CricKey technique. METHODS: This was a prospective crossover study using human cadaveric models. Participants included US Army combat medics credentialed at the emergency medical technician-basic level. After a brief anatomy review and demonstration, participants performed in random order standard open surgical cricothyroidotomy and CricKey surgical cricothyroidotomy. The primary outcome was first-pass success, and the secondary outcome measure was procedural time. RESULTS: First-attempt success was 100% (15/15) for CricKey surgical cricothyroidotomy and 66% (10/15) for open surgical cricothyroidotomy (odds ratio 16.0; 95% confidence interval 0.8 to 326). Surgical cricothyroidotomy insertion was faster for CricKey than open technique (34 versus 65 seconds; median time difference 28 seconds; 95% confidence interval 16 to 48 seconds). CONCLUSION: Compared with the standard open surgical cricothyroidotomy technique, military medics demonstrated faster insertion with the CricKey. First-pass success was not significantly different between the techniques.


Asunto(s)
Medicina Militar/métodos , Traqueostomía/métodos , Cadáver , Cartílago Cricoides/cirugía , Estudios Cruzados , Humanos , Medicina Militar/instrumentación , Factores de Tiempo , Traqueostomía/instrumentación
16.
Clin Chem ; 59(7): 1045-51, 2013 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-23509109

RESUMEN

BACKGROUND: Dried blood spot (DBS) samples have been widely used in newborn screening (NBS) for the early identification of disease to facilitate the presymptomatic treatment of congenital diseases in newborns. As molecular genetics knowledge and technology progresses, there is an increased demand on NBS programs for molecular testing and a need to establish reliable, low-cost methods to perform those analyses. Here we report a flexible, cost-efficient, high-throughput DNA extraction method from DBS adaptable to small- and large-scale screening settings. METHODS: Genomic DNA (g.DNA) was extracted from single 3-mm diameter DBS by the sequential use of red cell lysis, detergent-alkaline, and acid-neutralizing buffers routinely used in whole blood and plant tissue DNA extractions. We performed PCR amplification of several genomic regions using standard PCR conditions and detection methods (agarose gel, melting-curve analysis, TaqMan-based assays). Amplicons were confirmed by BigDye® Terminator cycle sequencing and compared with reference sequences. RESULTS: High-quality g.DNA was extracted from hundreds of DBS, as proven by mutation detection of several human genes on multiple platforms. Manual and automated extraction protocols were validated. Quantification of g.DNA by Oligreen® fluorescent nucleic acid stain demonstrated a normal population distribution closely corresponding with white blood cell counts detected in newborn populations. CONCLUSIONS: High-quality, amplifiable g.DNA is extractable from DBSs. Our method is adaptable, reliable, and scalable to low- and high-throughput NBS at low cost ($0.10/sample). This method is routinely used for molecular testing in the New York State NBS program.


Asunto(s)
ADN/aislamiento & purificación , Pruebas con Sangre Seca/métodos , Análisis Costo-Beneficio , ADN/sangre , Pruebas con Sangre Seca/economía , Humanos , Recién Nacido , Reacción en Cadena en Tiempo Real de la Polimerasa
17.
J Appl Lab Med ; 8(4): 674-688, 2023 07 05.
Artículo en Inglés | MEDLINE | ID: mdl-37207688

RESUMEN

BACKGROUND: Assessment of trace and toxic element status is important for the diagnosis and monitoring of several pediatric conditions. Elemental deficiency and toxicity have serious implications, particularly in pediatrics wherein risk is higher. Pediatric reference intervals (RIs) for trace elements and normal exposure limits for toxic elements are lacking on modern analytical systems. Herein, reference values were established for 13 plasma and 22 whole blood trace elements in the Canadian Laboratory Initiative on Pediatric Reference Intervals (CALIPER) cohort of healthy children and adolescents. METHODS: Approximately 320 healthy children and adolescents were recruited with informed consent. Trace elements were measured in whole blood and plasma samples using 2 technologies: (a) triple quadrupole inductively coupled plasma tandem mass spectrometry (ICP-MS/MS) (n = 172) and (b) high-resolution sector field ICPMS (HR-SF-ICPMS) (n =161). RIs and normal exposure limits were then established according to Clinical and Laboratory Standards Institute guidelines. RESULTS: Of all elements assessed, none required sex partitioning and 8 required age partitioning (e.g., copper, manganese, and cadmium). Reference value distributions determined via ICP-MS/MS and HR-SF-ICPMS demonstrated excellent concordance, with few exceptions (e.g., molybdenum, cobalt, and nickel). CONCLUSIONS: These data represent the first study wherein pediatric RIs and normal exposure limits were derived simultaneously on 2 different clinically validated MS platforms which provide urgently needed data to inform clinical decision-making for trace elements in pediatrics. Study findings suggest some trace elements require age-specific consideration for appropriate interpretation. Highly concordant observations across the 2 analytical methods also demonstrate the comparability and reliability of results obtained on both platforms.


Asunto(s)
Servicios de Laboratorio Clínico , Oligoelementos , Humanos , Niño , Adolescente , Oligoelementos/análisis , Valores de Referencia , Espectrometría de Masas en Tándem , Reproducibilidad de los Resultados
18.
J Appl Lab Med ; 8(5): 856-870, 2023 09 07.
Artículo en Inglés | MEDLINE | ID: mdl-37473432

RESUMEN

BACKGROUND: Vitamin D supplementation is common practice for neonates and infants due to limited stores of vitamin D at birth. Although not commonly encountered, vitamin D toxicity can occur due to over-supplementation. However, toxic concentrations are often not included in method validation experiments, and assays often are not validated in the neonatal population. METHODS: We compared serial 25 hydroxy vitamin D [25(OH)D] measurements in pre-term neonates receiving 25(OH)D supplementation and identified 12 patients wherein concentrations of 25(OH)D were above 50 ng/mL (125 nM) that required additional investigations as the 25(OH)D results did not match the clinical picture. Available samples were compared across 4 immunoassay platforms (LIAISON XL, Roche Cobas e602, Abbott Alinity i, and Siemens Centaur XP) and LC-MS/MS. RESULTS: Concentrations of 25(OH)D observed on one individual immunoassay platform (LIAISON XL) fluctuated substantially between subsequent blood draws in select neonates with elevated concentrations. Serum samples from these patients showed variable agreement between LC-MS/MS and other immunoassay platforms. These fluctuations were not explained by the presence of 3-epimer-25(OH)D or 24,25(OH)2D. CONCLUSIONS: Although we were unable to identify a cause for the variable elevated results, our findings suggest that neonatal 25(OH)D measurements alone should not be used for assessment of nutritional monitoring, and that clinical correlation and other laboratory parameters including ionized calcium should be considered.


Asunto(s)
Espectrometría de Masas en Tándem , Vitamina D , Recién Nacido , Humanos , Cromatografía Liquida/métodos , Espectrometría de Masas en Tándem/métodos , Inmunoensayo/métodos , Laboratorios
19.
J Appl Lab Med ; 7(2): 575-581, 2022 03 02.
Artículo en Inglés | MEDLINE | ID: mdl-34791316

RESUMEN

INTRODUCTION: A patient presented to hospital with chest pain and shortness of breath on 2 occasions 4 weeks apart. Clinical examination revealed an elevated jugular venous pressure consistent with heart failure or elevated filling pressures. METHODS: The patient was investigated through various modalities including electrocardiogram (ECG), transthoracic echocardiogram, coronary angiography, MRI, cardiac catheterization, positron emission tomography, and an extensive laboratory workup. RESULTS: Serial hs TnI measurements consistently revealed grossly elevated troponin I (>10 000 ng/L). In-lab investigation of increased high sensitivity troponin I (hsTnI) showed evidence of falsely increased troponin due to the presence of heterophilic antibodies. DISCUSSION: This case demonstrates a complex patient presentation and the value of involving the laboratory medicine team when dealing with potentially discrepant results. This is a rare report of grossly elevated troponin due to heterophilic antibodies for high-sensitivity troponin Abbott assay.


Asunto(s)
Dolor en el Pecho , Troponina I , Cateterismo Cardíaco , Dolor en el Pecho/diagnóstico , Dolor en el Pecho/etiología , Disnea/diagnóstico , Disnea/etiología , Humanos
20.
Curr Biol ; 32(15): 3334-3349.e6, 2022 08 08.
Artículo en Inglés | MEDLINE | ID: mdl-35797998

RESUMEN

Associative memory formation and recall in the fruit fly Drosophila melanogaster is subserved by the mushroom body (MB). Upon arrival in the MB, sensory information undergoes a profound transformation from broadly tuned and stereotyped odorant responses in the olfactory projection neuron (PN) layer to narrowly tuned and nonstereotyped responses in the Kenyon cells (KCs). Theory and experiment suggest that this transformation is implemented by random connectivity between KCs and PNs. However, this hypothesis has been challenging to test, given the difficulty of mapping synaptic connections between large numbers of brain-spanning neurons. Here, we used a recent whole-brain electron microscopy volume of the adult fruit fly to map PN-to-KC connectivity at synaptic resolution. The PN-KC connectome revealed unexpected structure, with preponderantly food-responsive PN types converging at above-chance levels on downstream KCs. Axons of the overconvergent PN types tended to arborize near one another in the MB main calyx, making local KC dendrites more likely to receive input from those types. Overconvergent PN types preferentially co-arborize and connect with dendrites of αß and α'ß' KC subtypes. Computational simulation of the observed network showed degraded discrimination performance compared with a random network, except when all signal flowed through the overconvergent, primarily food-responsive PN types. Additional theory and experiment will be needed to fully characterize the impact of the observed non-random network structure on associative memory formation and recall.


Asunto(s)
Drosophila melanogaster , Cuerpos Pedunculados , Animales , Drosophila/fisiología , Cuerpos Pedunculados/fisiología , Neuronas/fisiología , Olfato/fisiología
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