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1.
J Pharm Biomed Anal ; 243: 116082, 2024 Jun 15.
Article in English | MEDLINE | ID: mdl-38461636

ABSTRACT

BACKGROUND: Venlafaxine (VEN) and its O-demethylated metabolite, O-desmethylvenlafaxine (ODV), are commonly prescribed serotonin-norepinephrine reuptake inhibitors, approved for the treatment of depression and anxiety. Both are metabolized to inactive metabolites via cytochrome P450 enzymes. While previous studies have focused on quantifying VEN and ODV, bioanalytical methods for the simultaneous measurement of all metabolites are needed to fully characterize the pharmacology of VEN and ODV. METHODS: K2EDTA plasma was spiked with VEN, ODV, N-desmethylvenlafaxine (NDV), N,O-didesmethylvenlafaxine (NODDV), and N,N-didesmethylvenlafaxine (NNDDV). Drugs and metabolites were extracted via protein precipitation and quantified using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The multiplexed assay was validated in accordance with regulatory recommendations, and evaluated in remnant plasma samples from persons prescribed venlafaxine. RESULTS: The analytical measuring range for venlafaxine and all four metabolites was 5-800 ng/mL. Standard curves were generated via weighted quadratic (NNDDV) or linear (VEN, ODV, NDV, NODDV) regression of calibrators. Inter-assay imprecision was between 1.9-9.3% for all levels of all analytes. Minor matrix effects were observed, and both recovery efficiency and process efficiency were >96% for all analytes. All other assay validation assessments met acceptance criteria. Drug concentrations measured from remnant plasma specimens obtained from patients with current venlafaxine prescriptions (37.5-450 mg/day) yielded NDDV, NDV, and NODDV metabolite concentrations in 6/21, 14/21, and 20/21 samples, respectively. The ratio of active to inactive analytes ranged from 0.74 to 14.5, with a median of 6.39. CONCLUSIONS: An efficient and accurate LC-MS/MS method was developed and validated for the quantification of VEN, ODV, and all three inactive metabolites in plasma. The assay met all acceptance criteria, and may be used in future studies of the pharmacokinetics of these drugs.


Subject(s)
Cyclohexanols , Tandem Mass Spectrometry , Humans , Venlafaxine Hydrochloride , Chromatography, Liquid/methods , Tandem Mass Spectrometry/methods , Cyclohexanols/chemistry , Cyclohexanols/pharmacokinetics , Desvenlafaxine Succinate , Selective Serotonin Reuptake Inhibitors
2.
BMC Pulm Med ; 23(1): 442, 2023 Nov 15.
Article in English | MEDLINE | ID: mdl-37968677

ABSTRACT

BACKGROUND: The SARS-CoV2 pandemic impacted many critically ill patients, causing sequelae, affecting lung function, and involving the musculoskeletal system. We evaluated the association between lung function and muscle quality index in severely ill post-COVID-19 patients. METHODS: A cross-sectional study was conducted on a post-COVID-19 cohort at a third-level center. The study included patients who had experienced severe-to-critical COVID-19. Anthropometric measurements, such as body mass index (BMI) and handgrip strength, were obtained to calculate the muscle quality index (MQI). Additionally, spirometry, measurements of expiratory and inspiratory pressure, and an assessment of DLCO in the lungs were performed. The MQI was categorized into two groups: low-MQI (below the 50th percentile) and high-MQI (above the 50th percentile), based on sex. Group differences were analyzed, and a multivariate linear regression analysis was performed to assess the association between respiratory function and MQI. RESULTS: Among the 748 patients analyzed, 61.96% required mechanical ventilation, and the median hospital stay was 17 days. In patients with a low MQI, it was observed that both mechanical respiratory function and DLCO were lower. The multivariate analysis revealed significantly lower findings in mechanical respiratory function among patients with a low MQI. CONCLUSION: The Low-MQI is an independent predictor associated with pulmonary function parameters in subjects with Post-COVID-19 syndrome.


Subject(s)
COVID-19 , Musculoskeletal System , Humans , Hand Strength/physiology , Cross-Sectional Studies , Post-Acute COVID-19 Syndrome , RNA, Viral , SARS-CoV-2 , Lung , Muscles
3.
Bone ; 163: 116504, 2022 10.
Article in English | MEDLINE | ID: mdl-35878747

ABSTRACT

OBJECTIVES: We investigated the utility of urine phosphoethanolamine (PEA) as a marker to aid in diagnosing and/or confirming hypophosphatasia (HPP) in adults and for monitoring patients on enzyme replacement therapy (ERT). METHODS: Data was collected from seventy-eight adults who were referred to the Vanderbilt Program for Metabolic Bone Disease for evaluation of a possible or confirmatory HPP diagnosis between July 2014 through December 2019. Fifty-nine patients were diagnosed with HPP and nineteen were excluded from a diagnosis of HPP. The urine PEA results of those patients with a confirmed diagnosis of HPP and those patients with a diagnosis of HPP excluded were captured and compared to other laboratory and clinical parameters consistent with HPP, including alkaline phosphatase (ALP) activity, plasma pyridoxal 5'-phosphate (PLP), the presence of musculoskeletal abnormalities, and genetic testing for pathogenic mutations in ALPL. RESULTS: Initial urine PEA values in patients in our HPP cohort and not on ERT were significantly higher (median = 150.0 nmol/mg creatinine, IQR = 82.0-202.0) compared patients in our HPP negative group (median 18.0 nmol/mg creatinine, IQR = 14.0-30.0, p < 0.0001) and higher than patients on ERT (median 65.0 nmol/mg creatinine, IQR = 45.3-79.8). Patients who began ERT had a decline in urine PEA levels after treatment with a mean decrease of 68.1 %. Plasma ALP levels were significantly lower in the group of patients with HPP and not on ERT group (median = 24.0 U/L, IQR = 15.0-29.50) compared to the patients without HPP (median = 45.50 U/L, IQR = 34.0-62.0;) and plasma PLP levels were significantly higher in the HPP non-ERT group (median = 284.0 nmol/L, IQR = 141.0-469.4) compared to the patients without HPP (median = 97.5 nmol/L, IQR = 43.7-206.0;). The area under the curve (AUC) of urine PEA, ALP, and PLP to distinguish between HPP and non-HPP patients is 0.968, 0.927 and 0.781, respectively, in our cohort. Urine PEA had 100 % specificity (95 % CI of 83.2 % to 100.0 %) for diagnosing HPP at a value >53.50 nmol/mg creatinine with a sensitivity of 88.4 %; 95%CI 75.5 to 94.9 %. ALP had a 100 % specificity (95 % CI of 82.4 % to 100.0 %) for diagnosing HPP at a value <30.5 U/L with a sensitivity of 77.2 %; (95%CI 64.8 to 86.2 %). PLP had a 100 % specificity (95 % CI of 81.6 % to 100.0 %) for diagnosing HPP at a value >436 nmol/L with a sensitivity of 26.9 %; (95%CI 16.8 to 40.3 %). The most common pathogenic or likely pathogenic mutations in our cohort were c.1250A>G (p.Asn417Ser), c.1133A>T (p.Asp378Val), c.881A>C (p.Asp294Ala), c.1171C>T (p.Arg391Cys), and c.571G>A, (p.Glu191Lys). CONCLUSIONS: Urine PEA is a promising diagnostic and confirmatory marker for HPP in patients undergoing investigation for HPP. Urine PEA also has potential use as a marker to monitor ERT compliance. Future studies are necessary to evaluate the association between PEA levels and clinical outcomes.


Subject(s)
Hypophosphatasia , Adult , Alkaline Phosphatase , Biomarkers , Creatinine , Ethanolamines , Humans , Pyridoxal Phosphate
5.
J Anal Toxicol ; 46(1): 99-102, 2022 Feb 14.
Article in English | MEDLINE | ID: mdl-33216907

ABSTRACT

Point-of-care (POC) urine drug screening (UDS) assays provide immediate information for patient management. However, POC UDS assays can produce false-positive results, which may not be recognized until confirmatory testing is completed several days later. To minimize the potential for patient harm, it is critical to identify sources of interference. Here, we applied an approach based on statistical analysis of electronic health record (EHR) data to identify medications that may cause false positives on POC UDS assays. From our institution's EHR data, we extracted 120,670 POC UDS and confirmation results, covering 12 classes of target drugs, along with each individual's prior medication exposures. Our approach is based on the idea that exposure to an interfering medication will increase the odds of a false-positive UDS result. For a given assay-medication pair, we quantified the association between medication exposures and UDS results as an odds ratio from logistic regression. We evaluated interference experimentally by spiking compounds into drug-free urine and testing the spiked samples on the POC device. Our dataset included 446 false-positive UDS results (presumptive positive screen followed by negative confirmation). We quantified the odds ratio of false positives for 528 assay-medication pairs. Of the six assay-medication pairs we evaluated experimentally, two showed interference capable of producing a presumptive positive: labetalol on the 3,4-methylenedioxymethamphetamine (MDMA) assay (at 200 µg/mL) and ranitidine on the methamphetamine assay (at 50 µg/mL). Ranitidine also produced a presumptive positive for opiates at 1,600 µg/mL and for propoxyphene at 800 µg/mL. These findings highlight the generalizability and the limits of our approach to use EHR data to identify medications that interfere with clinical immunoassays.


Subject(s)
Electronic Health Records , Point-of-Care Systems , Substance Abuse Detection , Urinalysis , False Positive Reactions , Humans
6.
Microorganisms ; 11(1)2022 Dec 23.
Article in English | MEDLINE | ID: mdl-36677341

ABSTRACT

Phosphate-solubilizing bacteria (PSB) transform precipitated inorganic phosphorus into soluble orthophosphates. This study evaluated the efficiency of tricalcium and iron phosphate solubilization in Pikovskaya medium using five bacterial strains (A1, A2, A3, A5, and A6) cultured in acidic and alkaline pH levels. The bacterial strain that proved to be more efficient for P solubilization and was tolerant to pH variations was selected for assessing bacterial growth and P solubilization with glucose and sucrose in the culture medium. The bacterial strains were identified through 16S rRNA gene sequencing as Pseudomonas libanensis A1, Pseudomonas libanensis (A2), Bacillus pumilus (A3), Pseudomonas libanensis (A5), and Bacillus siamensis (A6). These five bacterial strains grew, tolerated pH changes, and solubilized inorganic phosphorus. The bacterial strain A3 solubilized FePO4 (4 mg L-1) and Ca3(PO4)2 (50 mg L-1). P solubilization was assayed with glucose and sucrose as carbon sources for A3 (Bacillus pumilus MN100586). After four culture days, Ca3(PO4)2 was solubilized, reaching 246 mg L-1 with sucrose in culture media. Using glucose as a carbon source, FePO4 was solubilized and reached 282 mg L-1 in six culture days. Our findings were: Pseudomonas libanensis, and Bacillus siamensis, as new bacteria, can be reported as P solubilizers with tolerance to acidic or alkaline pH levels. The bacterial strain B. pumilus grew using two sources of inorganic phosphorus and carbon, and it tolerated pH changes. For that reason, it is an ideal candidate for inorganic phosphorus solubilization and future production as a biofertilizer.

7.
SN Soc Sci ; 1(6): 140, 2021.
Article in English | MEDLINE | ID: mdl-34693322

ABSTRACT

Career development programs are a valuable part of any student's experience, and increasingly is an expected part of graduate school training. While such programs are commonly available to undergraduates, there is a growing need for career support to be offered to graduate students. Making the case for resources can be a challenge in this domain, however. Research on the impact of career services for graduate students and post-doctoral scholars is a growing scholarly concern. However, there remains a need to better understand what level of intervention is most appropriate: What kind of activities, how much time, and what resources would best serve the professional development needs of graduate students and post-doctoral scholars? And to answer these questions, a more foundational one: what activities are drawing the attention of graduate students and post doctoral trainees, and in what activities are they spending their time? In this manuscript, we describe how Our University approached this research question by developing an online data tracking system to capture graduate and post-doctoral trainee participation in one co-curricular professional development program. We demonstrate how this data tracking system can be used to advocate for institutional resources in career development programming, for research, and for practical purposes such as advocating for institutional support and for program design and assessment.

8.
Blood Transfus ; 19(4): 317-326, 2021 07.
Article in English | MEDLINE | ID: mdl-34059188

ABSTRACT

BACKGROUND: Following the first reports in the literature, the association between the ABO blood group and SARS-CoV-2 infection has been investigated by a number of studies, although with varying results. The main object of this systematic review was to assess the relationship between the ABO blood group and the occurrence and severity of COVID-19. MATERIALS AND METHODS: A systematic literature search using appropriate MeSH terms was performed through Medline and PubMed. The outcomes considered were the prevalence of the blood group O vs non-O types in SARS-CoV-2 infected and non-infected subjects, and the severity of SARS-CoV-2 infection according to ABO group. The methodological quality of the studies included in the analysis was assessed with the Newcastle-Ottawa Scale, and the overall quality of the available evidence using the GRADE system. Benchmarks used to evaluate the effect size were odd ratios (ORs) for case control studies and risk ratios (RRs) for cohort studies. RESULTS: Twenty-one studies were included in the analysis. Overall, individuals with group O had a lower infection rate compared to individuals of non-O group (OR: 0.81; 95% CI: 0.75, 0.86). However, the difference in the effect size was significantly lower in cohort studies compared to case control studies. No evidence was found indicating an effect of the O type on the disease severity in the infected patients. DISCUSSION: We have found low/very low evidence that group O individuals are less susceptible to SARS-CoV-2 infection compared to those in the non-O group. No evidence was found indicating an effect of the O type on disease severity in SARS-CoV-2 infection.


Subject(s)
ABO Blood-Group System , COVID-19/blood , Severity of Illness Index , Benchmarking , Case-Control Studies , Cohort Studies , Humans , Odds Ratio , SARS-CoV-2
9.
Compr Physiol ; 11(3): 2279-2295, 2021 06 30.
Article in English | MEDLINE | ID: mdl-34190339

ABSTRACT

Regulation of the ability of a neurotransmitter [our focus: serotonin, norepinephrine, dopamine, acetylcholine, glycine, and gamma-aminobutyric acid (GABA)] to reach its receptor targets is regulated in part by controlling the access the neurotransmitter has to receptors. Transporters, located at both the cellular plasma membrane and in subcellular vesicles, carry a myriad of responsibilities that include enabling neurotransmitter release and controlling uptake of neurotransmitter back into a cell or vesicle. Driven largely by electrochemical gradients, these transporters move neurotransmitters. The regulation of the transporters themselves through changes in expression and/or posttranslational modification allows for fine-tuning of this system. Transporters have been best recognized as targets for psychoactive stimulants and remain a mainstay target of primarily central nervous system (CNS) acting drugs for treatment of debilitating diseases such as depression and anxiety. Studies reveal, however, that transporters are found and functional in tissues outside the CNS (gastrointestinal and cardiovascular tissues, for example). The importance of neurotransmitter transporters is underscored with discoveries that dysfunction of transporters can cause life-changing disease. This article provides a high-level review of major classes of both plasma membrane transporters and vesicular transporters. © 2021 American Physiological Society. Compr Physiol 11:2279-2295, 2021.


Subject(s)
Membrane Transport Proteins , Neurotransmitter Transport Proteins , Biological Transport , Carrier Proteins/metabolism , Neurotransmitter Agents , Neurotransmitter Transport Proteins/genetics , Neurotransmitter Transport Proteins/metabolism
10.
Clin Chim Acta ; 520: 118-125, 2021 Sep.
Article in English | MEDLINE | ID: mdl-34043979

ABSTRACT

BACKGROUND: Routine chemistry testing is typically performed using serum or plasma to assess a patient's clinical status. At our institution, serum is the specimen type used. To reduce processing times, evaluation of plasma-based and rapid serum gel separator tubes was performed. METHODS: We compared the results of routine chemistry analytes collected in serum gel separator tubes (SST), plasma gel separator tubes (PST), rapid serum gel separator tubes (RST), and plasma tubes without gel separators (DGT). Result concordance was assessed at baseline (immediate testing after processing) and up to one week of refrigerated storage. Other parameters assessed were the susceptibility to hemolysis and lipemia interference, and changes in results after re-centrifugation. Percent changes were compared against the SST and evaluated according to established bias thresholds. RESULTS: Total protein and potassium results at baseline in plasma-based tubes had percent changes from the SST that exceeded acceptability thresholds. Stability was significantly shortened for glucose, potassium, aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) when collected in the PST as compared to the SST. The RST was the least susceptible to hemolysis and lipemia interferents. Re-centrifugation affected the serum-based analysis of potassium. CONCLUSIONS: Plasma may reduce processing time at the expense of shortened sample stability and may require specimen source-specific reference intervals for potassium and total protein. The RST provides an alternate option to reduce processing time, while maintaining storage stability.


Subject(s)
Blood Specimen Collection , Chemistry, Clinical , Humans , Plasma , Potassium , Serum
11.
Arch Cardiol Mex ; 91(4): 415-421, 2021 Nov 01.
Article in Spanish | MEDLINE | ID: mdl-33401285

ABSTRACT

Antecedents and objective: The knowledge of the reference levels for diagnoses and therapeutic studies is important, because it contributes to the optimization of the radiological protection of the patients, avoiding them to be exposed to unnecessarily high doses. However, there was no evidence of these levels in interventionist cardiology procedures in Ecuador, so the objective of this study was to estimate the diagnostic reference levels of dose at the entrance surface in adult patients undergoing interventional cardiology procedures (cinecoronariography, catheterization and coronary percutaneous interventionism) in the Unit of Hemodynamic of the Hospital of Specialties Carlos Andrade Marín in Ecuador. Materials and methods: The measurements of product dose-area, dose in input surface, number of images and the time of fluoroscopy performed using the Axiom Artis angiographer, and the obtained data was tabulated and processed with the Excel computer program. For this study, 145 patients older than 18 years were selected, and the values obtained from diagnostic reference dose levels were compared with others reported in previous investigations and with the permissible limits for the appearance of deterministic effects on the skin. Results: This study made it possible to establish reference levels for diagnosis in interventional cardiology procedures (coronary angiography, catheterization and percutaneous coronary intervention) in the Hemodynamics Unit of the Carlos Andrade Marín Specialty Hospital in Ecuador. Conclusions: The results showed that the intake surface doses obtained were lower than the maximum levels recommended by the International Atomic Energy Agency.


Antecedentes y objetivo: El conocimiento de los niveles de referencia para estudios diagnósticos y terapéuticos es importante, dado que contribuye a la optimización de la protección radiológica de los pacientes y evita que se expongan a dosis innecesariamente altas; sin embargo, no se encontraron evidencias de estos niveles en procedimientos de cardiología intervencionista en Ecuador, por lo cual el objetivo de este estudio fue calcular los niveles de referencia diagnósticos de dosis en la superficie de entrada en pacientes adultos sometidos a procedimientos intervencionistas de cardiología (cinecoronariografía, cateterismo e intervencionismo percutáneo coronario) en la Unidad de Hemodinámica del Hospital de Especialidades Carlos Andrade Marín de Ecuador. Materiales y métodos: Las mediciones del producto dosis-área, dosis en superficie de entrada, número de imágenes y tiempo de fluoroscopia se realizaron con el angiógrafo Axiom Artis y los datos obtenidos se tabularon y procesaron con el programa informático Excel. Para este estudio se seleccionó a 145 pacientes mayores de 18 años y los valores obtenidos de niveles de referencia diagnósticos de dosis se compararon con otros informados en investigaciones anteriores y con los límites permisibles para la aparición de efectos deterministas en piel. Resultados: Este estudio permitió establecer los niveles de referencia para diagnóstico en los procedimientos intervencionistas de cardiología (cinecoronariografía, cateterismo e intervencionismo percutáneo coronario) en la Unidad de Hemodinámica del Hospital de Especialidades Carlos Andrade Marín de Ecuador. Conclusiones: Los resultados mostraron que las dosis en superficie de entrada obtenidas fueron menores a los niveles máximos recomendados por la International Atomic Energy Agency.


Subject(s)
Cardiology/methods , Adult , Aged , Aged, 80 and over , Cardiology/standards , Catheterization , Cineangiography , Ecuador , Female , Humans , Magnetic Resonance Imaging, Cine , Male , Middle Aged , Percutaneous Coronary Intervention , Radiation Dosage , Radiation Protection/methods , Radiography, Interventional/methods , Reference Standards , Reference Values
12.
J Appl Lab Med ; 6(1): 319-323, 2021 01 12.
Article in English | MEDLINE | ID: mdl-33438742
13.
Am J Clin Pathol ; 155(4): 537-546, 2021 03 15.
Article in English | MEDLINE | ID: mdl-33210111

ABSTRACT

OBJECTIVES: Despite extensive research on procalcitonin (PCT)-guided therapy in lower respiratory tract infections, the association between PCT and bacterial pneumonia remains unclear. METHODS: We evaluated retrospectively the performance of PCT in patients presenting with lower respiratory tract infection symptoms and grouped by seven diagnoses. All patients had microbial testing, chest imaging, and CBC counts within 1 day of PCT testing. RESULTS: Median PCT level in patients diagnosed with bacterial pneumonia was significantly higher than in patients diagnosed with other sources of infections or those not diagnosed with infections. Median PCT levels were not different among patients grouped by type or quantity of pathogen detected. They were significantly higher in patients with higher pathogenicity scores for isolated bacteria, those with abnormal WBC count, and those with chest imaging consistent with bacterial pneumonia. A diagnostic workup that included imaging, WBC count, and Gram stain had an area under the receiver operating characteristic curve of 0.748, and the addition of PCT increased it to 0.778. CONCLUSIONS: PCT was higher in patients diagnosed with bacterial pneumonia. Less clear is its diagnostic ability to detect bacterial pneumonia over and above imaging and laboratory data routinely available to clinicians.


Subject(s)
Biomarkers/blood , Pneumonia, Bacterial/blood , Pneumonia, Bacterial/diagnosis , Procalcitonin/blood , Adult , Aged , Female , Humans , Male , Middle Aged , Retrospective Studies
14.
J Anal Toxicol ; 45(4): 325-330, 2021 Apr 12.
Article in English | MEDLINE | ID: mdl-32991692

ABSTRACT

Urine drug screening (UDS) assays can rapidly and sensitively detect drugs of abuse but can also produce spurious results due to interfering substances. We previously developed an approach to identify interfering medications using electronic health record (EHR) data, but the approach was limited to UDS assays for which presumptive positives were confirmed using more specific methods. Here we adapted the approach to search for medications that cause false positives on UDS assays lacking confirmation data. From our institution's EHR data, we used our previous dataset of 698,651 UDS and confirmation results. We also collected 211,108 UDS results for acetaminophen, ethanol and salicylates. Both datasets included individuals' prior medication exposures. We hypothesized that the odds of a presumptive positive would increase following exposure to an interfering medication independently of exposure to the assay's target drug(s). For a given assay-medication pair, we quantified potential interference as an odds ratio from logistic regression. We evaluated interference of selected compounds in spiking experiments. Compared to the approach requiring confirmation data, our adapted approach showed only modestly diminished ability to detect interfering medications. Applying our approach to the new data, we discovered and validated multiple compounds that can cause presumptive positives on the UDS assay for acetaminophen. Our approach can reveal interfering medications using EHR data from institutions at which UDS results are not routinely confirmed.


Subject(s)
Substance Abuse Detection , Drug Evaluation, Preclinical , Humans
15.
Arch Pathol Lab Med ; 144(10): 1193-1198, 2020 10 01.
Article in English | MEDLINE | ID: mdl-33002154

ABSTRACT

CONTEXT.­: Convenience, avoidance of doctor's appointments, curiosity, and the desire to take control of one's health are driving interest toward direct-to-consumer (DTC) testing. DTC is laboratory testing that is initiated by the consumer without a physician order. The results are reported back directly to the consumer. DTC testing is an exciting addition to the traditional healthcare model for consumers who want knowledge of their health status and disease risk, ancestry, and their body's expected response to certain medications based on their genotype. OBJECTIVES.­: To discuss the perceived and potential benefits and risks involved in DTC testing. DATA SOURCES.­: Recent published literature on DTC testing. CONCLUSIONS.­: The benefits of DTC testing are enticing and are driving the DTC testing market. Consumers must weigh the perceived benefits with the potential risks, including privacy concerns, the possibility of receiving confusing health information, and/or information that could generate unexpected emotions, misdiagnosis, and over-testing.


Subject(s)
Direct-To-Consumer Screening and Testing , Confidentiality , Genetic Testing , Humans , Risk Assessment
16.
Blood ; 136(23): 2691-2702, 2020 12 03.
Article in English | MEDLINE | ID: mdl-32659785

ABSTRACT

The mechanisms by which phlebotomy promotes the mobilization of hepatic iron stores are not well understood. NCOA4 (nuclear receptor coactivator 4) is a widely expressed intracellular protein previously shown to mediate the autophagic degradation of ferritin. Here, we investigate a local requirement for NCOA4 in the regulation of hepatic iron stores and examine mechanisms of NCOA4 regulation. Hepatocyte-targeted Ncoa4 knockdown in nonphlebotomized mice had only modest effects on hepatic ferritin subunit levels and nonheme iron concentration. After phlebotomy, mice with hepatocyte-targeted Ncoa4 knockdown exhibited anemia and hypoferremia similar to control mice with intact Ncoa4 regulation but showed a markedly impaired ability to lower hepatic ferritin subunit levels and hepatic nonheme iron concentration. This impaired hepatic response was observed even when dietary iron was limited. In both human and murine hepatoma cell lines, treatment with chemicals that stabilize hypoxia inducible factor (HIF), including desferrioxamine, cobalt chloride, and dimethyloxalylglycine, raised NCOA4 messenger RNA. This NCOA4 messenger RNA induction occurred within 3 hours, preceded a rise in NCOA4 protein, and was attenuated in the setting of dual HIF-1α and HIF-2α knockdown. In summary, we show for the first time that NCOA4 plays a local role in facilitating iron mobilization from the liver after blood loss and that HIF regulates NCOA4 expression in cells of hepatic origin. Because the prolyl hydroxylases that regulate HIF stability are oxygen- and iron-dependent enzymes, our findings suggest a novel mechanism by which hypoxia and iron deficiency may modulate NCOA4 expression to impact iron homeostasis.


Subject(s)
Basic Helix-Loop-Helix Transcription Factors/metabolism , Hemorrhage/metabolism , Hepatocytes/metabolism , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Iron/metabolism , Liver/metabolism , Nuclear Receptor Coactivators/biosynthesis , Animals , Basic Helix-Loop-Helix Transcription Factors/genetics , Cell Line , Female , Gene Expression Regulation , Gene Knockdown Techniques , Hemorrhage/genetics , Hemorrhage/pathology , Hepatocytes/pathology , Humans , Hypoxia-Inducible Factor 1, alpha Subunit/genetics , Liver/pathology , Mice , Nuclear Receptor Coactivators/genetics
17.
J Appl Lab Med ; 5(1): 126-135, 2020 01 01.
Article in English | MEDLINE | ID: mdl-32445342

ABSTRACT

BACKGROUND: The anion gap is primarily used in the diagnosis of acid-base disorders. We conducted a study to determine the anion gap reference interval in our patient population, investigated the workup of abnormal vs normal anion gaps, and examined the anion gap variation upon repeated testing. METHODS: A retrospective review was performed on 17137 adult and pediatric patients who presented to Yale-New Haven Hospital outpatient clinics, emergency department, or intensive care units between 2012 and 2017. RESULTS: We derived a new reference interval of 7 to 18 mmol/L with a median of 13 mmol/L in healthy adults with no significant differences owing to partitioning by sex or age. Based on the new reference interval, 5%, 23%, and 18% of healthy, emergency department, and intensive care unit adult patients, respectively, were misclassified as having high values with the previous interval of 6 to 16 mmol/L. However, there were no significant differences in the number of tests ordered in patients with anion gaps above and below the upper limit of the previous reference interval. The majority of increased anion gaps that were repeated normalized by 12 h. In a subgroup of healthy adult patients with annual testing, the median percent change in each patient's anion gap from 2015 to 2016 was approximately 13%. CONCLUSIONS: The anion gap should be used with an appropriate reference interval to avoid misclassification. There may be a moderate degree of individuality that argues for comparing the anion gap with its baseline value in the same patient pending further studies that formally derive its biological variation.


Subject(s)
Acid-Base Equilibrium/physiology , Acid-Base Imbalance , Acid-Base Imbalance/blood , Acid-Base Imbalance/diagnosis , Adult , Age Factors , Analysis of Variance , Biological Variation, Population , Child , Female , Humans , Male , Reference Values , Retrospective Studies
20.
Pol J Microbiol ; 69(3): 357-365, 2020 Sep.
Article in English | MEDLINE | ID: mdl-33574865

ABSTRACT

The capacity of four bacterial strains isolated from productive soil potato fields to solubilize tricalcium phosphate on Pikovskaya agar or in a liquid medium was evaluated. A bacterial strain was selected to evaluate in vitro capacity of plant-growth promotion on Solanum tuberosum L. culture. Bacterial strain A3 showed the highest value of phosphate solubilization, reaching a 20 mm-diameter halo and a concentration of 350 mg/l on agar and in a liquid medium, respectively. Bacterial strain A3 was identified by 16S rDNA analysis as Bacillus pumilus with 98% identity; therefore, it is the first report for Bacillus pumilus as phosphate solubilizer. Plant-growth promotion assayed by in vitro culture of potato microplants showed that the addition of bacterial strain A3 increased root and stems length after 28 days. It significantly increased stem length by 79.3%, and duplicated the fresh weight of control microplants. In this paper, results reported regarding phosphorus solubilization and growth promotion under in vitro conditions represent a step forward in the use of innocuous bacterial strain biofertilizer on potato field cultures.


Subject(s)
Bacteria/metabolism , Phosphates/metabolism , Soil Microbiology , Solanum tuberosum/growth & development , Bacillus pumilus/classification , Bacillus pumilus/genetics , Bacillus pumilus/isolation & purification , Bacillus pumilus/metabolism , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , Kinetics , Phylogeny , Plant Roots/growth & development , Plant Stems/growth & development , RNA, Ribosomal, 16S/genetics , Rhizosphere , Solanum tuberosum/metabolism , Sucrose/metabolism
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