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International Journal of Neonatal... Feb 2024Very-long-chain acyl-CoA dehydrogenase (VLCAD) deficiency is a long-chain fatty acid oxidation disorder that manifests as either a severe phenotype associated with...
Using the C14:1/Medium-Chain Acylcarnitine Ratio Instead of C14:1 to Reduce False-Positive Results for Very-Long-Chain Acyl-CoA Dehydrogenase Deficiency in Newborn Screening in Japan.
Very-long-chain acyl-CoA dehydrogenase (VLCAD) deficiency is a long-chain fatty acid oxidation disorder that manifests as either a severe phenotype associated with cardiomyopathy, a hypoglycemic phenotype, or a myopathic phenotype. As the hypoglycemic phenotype can cause sudden infant death, VLCAD deficiency is included in newborn screening (NBS) panels in many countries. The tetradecenoylcarnitine (C14:1) level in dried blood specimens is commonly used as a primary marker for VLCAD deficiency in NBS panels. Its ratio to acetylcarnitine (C2) and various other acylcarnitines is used as secondary markers. In Japan, tandem mass spectrometry-based NBS, initially launched as a pilot study in 1997, was introduced to the nationwide NBS program in 2013. In the present study, we evaluated levels of acylcarnitine with various chain lengths (C18 to C2), free carnitine, and their ratios in 175 infants who tested positive for VLCAD deficiency with C14:1 and C14:1/C2 ratios. Our analyses indicated that the ratios of C14:1 to medium-chain acylcarnitines (C10, C8, and C6) were the most effective markers in reducing false-positive rates. Their use with appropriate cutoffs is expected to improve NBS performance for VLCAD deficiency.
PubMed: 38390979
DOI: 10.3390/ijns10010015 -
The Journal of Applied Laboratory... May 2024In addition to newborn screening, dried blood spots (DBSs) are used for a wide variety of analytes for clinical, epidemiological, and research purposes. Guidelines on...
BACKGROUND
In addition to newborn screening, dried blood spots (DBSs) are used for a wide variety of analytes for clinical, epidemiological, and research purposes. Guidelines on DBS collection, storage, and transport are available, but it is suggested that each laboratory should establish its own acceptance criteria.
METHODS
An optical scanning device was developed to assess the quality of DBSs received in the newborn screening laboratory from 11 maternity wards between 2013 and 2018. The algorithm was adjusted to agree with the visual examination consensus of experienced laboratory personnel. Once validated, the algorithm was used to categorize DBS specimens as either proper or improper. Improper DBS specimens were further divided based on 4 types of specimen defects.
RESULTS
In total, 27 301 DBSs were analyzed. Compared with an annual DBS rejection rate of about 1%, automated scanning rejected 26.96% of the specimens as having at least one defect. The most common specimen defect was multi-spotting (ragged DBS, 19.13%). Among maternity wards, improper specimen rates varied greatly between 5.70% and 49.92%.
CONCLUSIONS
Improper specimen rates, as well as the dominant type of defect(s), are mainly institution-dependent, with various maternity wards consistently showing specific patterns of both parameters over time. Although validated in agreement with experienced laboratory personnel consensus, automated analysis rejects significantly more specimens. While continuous staff training, specimen quality monitoring, and problem-reporting to maternities is recommended, a thorough quality assessment strategy should also be implemented by every newborn screening laboratory. An important role in this regard may be played by automation in the form of optical scanning devices.
Topics: Humans; Neonatal Screening; Infant, Newborn; Dried Blood Spot Testing; Algorithms; Quality Assurance, Health Care; Quality Control; Blood Specimen Collection
PubMed: 38384160
DOI: 10.1093/jalm/jfae003 -
Journal of Medical Entomology Feb 2024Chagas disease, caused by the protozoan Trypanosoma cruzi, is a zoonosis primarily found in rural areas of Latin America. It is considered a neglected tropical disease,...
Chagas disease, caused by the protozoan Trypanosoma cruzi, is a zoonosis primarily found in rural areas of Latin America. It is considered a neglected tropical disease, and Triatoma dimidiata is the main vector of the parasite in Central America. Despite efforts, Chagas disease continues to be a public health concern, and vector control remains a primary tool to reduce transmission. In this study, we tested the hypothesis that highly abundant bacteria in the gut of T. dimidiata inhibit the growth of T. cruzi. To achieve this, bacterial diversity in the gut of T. dimidiata specimens from Costa Rica was characterized by metabarcoding of the 16S rRNA, microbial isolation was performed, and the effect of freeze-dried supernatants of the isolates on T. cruzi was investigated. Metabarcoding showed that the most abundant genera in the gut were Corynebacterium, Tsukamurella, Brevibacterium, and Staphylococcus. Barcoding and sequences comparison confirmed that 8 of the 30 most abundant amplicon sequence variants (ASVs) were isolated, and 2 of them showed an inhibitory effect on the growth of T. cruzi epimastigotes. These bacteria correspond to isolates of Tsukamurella and Brevibacterium, which were respectively the second and sixth most abundant ASVs in the gut of T. dimidiata. Notably, only the isolate of Brevibacterium showed a significant difference in growth inhibition against epimastigotes of both T. cruzi strains tested. These findings suggest that the gut microbiota of T. dimidiata may play an active role in modulating parasite development.
PubMed: 38381588
DOI: 10.1093/jme/tjae012 -
Journal of Food Science Apr 2024Persimmons contribute positively to human health. Although off-season utilization typically presents a challenge due to permissions' perishable nature, it may become...
Persimmons contribute positively to human health. Although off-season utilization typically presents a challenge due to permissions' perishable nature, it may become feasible through the implementation of appropriate drying methods. In this study, round sliced samples were dried to assess drying kinetics, modeling potential, color attributes, rehydration capacity, energy consumption (EC), cost index, and thermal properties. The fruits were subjected to distinct drying methodologies including freeze-drying, continuous infrared drying (300, 400, and 500 W), and intermittent infrared drying (PR = 1 [continuous], PR = 2 [30 s on-30 s off], and PR = 3 [20 s on-40 s off]). The duration of the drying process ranged from 40 to 390 min. It was determined that the most suitable models for depicting continuous and infrared drying kinetics of persimmon fruit were the Midilli et al. and Page models, whereas the Logarithmic model was identified as the optimal choice for characterization of freeze-drying kinetics. Assessment of EC revealed that both intermittent and continuous infrared drying methods incurred lower energy expenditure in comparison to the freeze-drying technique. Remarkably, throughout the course of the infrared drying processes, product surface temperatures varied between 106.33 and 22.65°C across different treatments. Despite its high EC, it has been found that high-quality products are produced by freeze-drying. However, infrared and intermittent infrared applications can be a low energy cost and feasible method for drying persimmon with a shorter duration. PRACTICAL APPLICATION: Persimmon is an important fruit with high nutritional value. However, as with many fresh products, they have a short shelf life. Within the scope of this research, three different drying methodologies were employed in the desiccation of persimmon specimens, and the impact of these methodologies on the overall qualitative attributes of the persimmon product was investigated. Despite its elevated energy consumption, the freeze-drying approach was found to yield high-quality products. Moreover, it was discerned that infrared drying represented a viable and expeditious alternative for drying the fruit, particularly when executed intermittently.
Topics: Humans; Desiccation; Diospyros; Fruit; Freeze Drying; Temperature
PubMed: 38380681
DOI: 10.1111/1750-3841.16994 -
Cancer Cytopathology May 2024Circulating tumor cells (CTCs) shed into blood provide prognostic and/or predictive information. Previously, the authors established an assay to detect carcinoma cells...
BACKGROUND
Circulating tumor cells (CTCs) shed into blood provide prognostic and/or predictive information. Previously, the authors established an assay to detect carcinoma cells from pleural fluid, termed effusion tumor cells (ETCs), by employing an immunofluorescence-based CTC-identification platform (RareCyte) on air-dried unstained ThinPrep (TP) slides. To facilitate clinical integration, they evaluated different slide processing and storage conditions, hypothesizing that alternative comparable conditions for ETC detection exist.
METHODS
The authors enumerated ETCs on RareCyte, using morphology and mean fluorescence intensity (MFI) cutoffs of >100 arbitrary units (a.u.) for epithelial cellular adhesion molecule (EpCAM) and <100 a.u. for CD45. They analyzed malignant pleural fluid from three patients under seven processing and/or staining conditions, three patients after short-term storage under three conditions, and seven samples following long-term storage at -80°C. MFI values of 4',6-diamidino-2-phenylindol, cytokeratin, CD45, and EpCAM were compared.
RESULTS
ETCs were detected in all conditions. Among the different processing conditions tested, the ethanol-fixed, unstained TP was most similar to the previously established air-dried, unstained TP protocol. All smears and Pap-stained TPs had significantly different marker MFIs from the established condition. After short-term storage, the established condition showed comparable results, but ethanol-fixed and Pap-stained slides showed significant differences. ETCs were detectable after long-term storage at -80°C in comparable numbers to freshly prepared slides, but most marker MFIs were significantly different.
CONCLUSIONS
It is possible to detect ETCs under different processing and storage conditions, lending promise to the application of this method in broader settings. Because of decreased immunofluorescence-signature distinctions between cells, morphology may need to play a larger role.
Topics: Humans; Neoplastic Cells, Circulating; Pleural Effusion, Malignant; Epithelial Cell Adhesion Molecule; Specimen Handling; Biomarkers, Tumor; Leukocyte Common Antigens; Fluorescent Antibody Technique
PubMed: 38373107
DOI: 10.1002/cncy.22803 -
Stomatologiia 2024The era of using adhesives to fix brackets began in the 70s of the last century. There are eight generations of foreign-made adhesive systems for the restoration of...
BACKGROUND
The era of using adhesives to fix brackets began in the 70s of the last century. There are eight generations of foreign-made adhesive systems for the restoration of teeth. However, until now, in orthodontics, the development of domestic adhesive systems with improved properties of adhesion of brackets to tooth enamel still important, especially in the posterior teeth.
OBJECTIVE
The purpose of this work was to study the shear strength of fifth generation domestic adhesive between metal brackets and the enamel of molar teeth in laboratory conditions.
MATERIAL AND METHODS
The study included 17 specimens of extracted maxillary molars embedded in acrylic resin blocks. Brackets from G&H Orthodontics (G&H Wire Company, USA) were fixed to the tooth enamel using the domestic Compofix (ortho) set. The enamel was treated with etching gel (37% phosphoric acid) for 30 seconds. Then the gel was washed off with water and the surface of the enamel was thoroughly dried. A primer was applied to the prepared surface with an applicator, inflated with a weak air flow for 5-10 seconds and photopolymerized for 20 seconds. The surface of the bracket was treated with degreaser, then a thin layer of adhesive was applied, and the sample was fixed to the enamel. Excess material was removed with an applicator. The samples were photopolymerized for 20 sec. Then, the shear bond strength of the adhesive was determined according to the method of GOST 31574-2012 on a (Zwick/Roell Z010 testing machine, Zwick, Germany).
RESULTS
The shear strength of the adhesive joint obtained during the test is 13.54±1.01 MPa, the average value of the index of adhesive residues on the surface of the bracket was 23%±4%, which corresponds to the standard average values according to GOST.
CONCLUSION
The tested domestic adhesive system of the fifth generation can be recommended for the practical work of an orthodontist.
Topics: Humans; Dental Bonding; Dental Cements; Dental Enamel; Molar; Russia; Orthodontic Brackets; Materials Testing; Surface Properties; Dental Stress Analysis; Stress, Mechanical
PubMed: 38372599
DOI: 10.17116/stomat20241030115 -
Molecules (Basel, Switzerland) Jan 2024Lam. is a thorny Iberian Peninsula endemic species belonging to the Apiaceae family that has not been previously analysed from a chemical point of view. Following our...
Lam. is a thorny Iberian Peninsula endemic species belonging to the Apiaceae family that has not been previously analysed from a chemical point of view. Following our studies on this genus, we characterized the chemical composition of the essential oils from the different parts (inflorescences, stems + leaves, and roots) of this species; these parts were gathered in Cádiz (Spain). The specimens were collected in July during the flowering period and air-dried before the oil extraction by hydro-distillation. The essential oils were analysed by gas chromatography and gas chromatography coupled with mass spectrometry. The different parts of the plant yielded low amounts of pale yellow oil, with the roots being the fraction that provided the lowest amount of oil. The chemical characterization of the essential oils showed qualitative and quantitative differences between the fractions examined, but all of them showed the same principal compound, germacrene D (9.1-46.5%). Similarly, all the fractions shared most of their representative constituents, with their percentage compositions being different from one sample to the other: α-cadinol (3.8%), bicyclogermacrene (3.5%), octanal (3.1%), and spathulenol (2.5%) were found in the inflorescences; octanal (8.1%), α-cadinol (3.7%), δ-cadinene (3.6%), ()-caryophyllene (2.6%), bicyclogermacrene (2.5%), and spathulenol (2.4%) were found in the stems and leaves; and spathulenol (4.6%), α-cadinol (4.4%), khusinol (3.2%), α-muurolol (3.1%), and δ-cadinene (2.6%) were found in the roots. As far as we know, this is the first report about the chemical composition of this endemic species of the Iberian Peninsula. It contributes to the knowledge of this species and to the genus to which it belongs. This species could be considered as a natural source of germacrene D, which is a sesquiterpene hydrocarbon with active properties.
Topics: Oils, Volatile; Eryngium; Gas Chromatography-Mass Spectrometry; Polycyclic Sesquiterpenes; Aldehydes; Sesquiterpenes, Germacrane; Terpenes; Sesquiterpenes
PubMed: 38338307
DOI: 10.3390/molecules29030562 -
Therapeutic Drug Monitoring Jan 2024Volumetric Absorptive Microsampling (VAMS) is a useful tool for therapeutic drug monitoring (TDM) of oral targeted anticancer agents. VAMS aims to improve safety and...
Analytical Validation of a Volumetric Absorptive Microsampling Method for Therapeutic Drug Monitoring of the Oral Targeted Anticancer Agents, Abiraterone, Alectinib, Cabozantinib, Imatinib, Olaparib, and Sunitinib, and Metabolites.
BACKGROUND
Volumetric Absorptive Microsampling (VAMS) is a useful tool for therapeutic drug monitoring (TDM) of oral targeted anticancer agents. VAMS aims to improve safety and efficacy by enabling at-home blood sample collection by patients. This study aimed to develop and validate an ultra-high performance liquid chromatography-tandem mass spectrometry method for the quantitative determination of abiraterone, alectinib, cabozantinib, imatinib, olaparib, sunitinib, and the metabolites, Δ(4)-abiraterone (D4A), alectinib-M4, imatinib-M1, and N-desethyl sunitinib, in dried whole blood samples using VAMS to support TDM.
METHODS
After the collection of 10 μL of whole blood sample using the VAMS device, the analytes were extracted from the tip using methanol with shaking, evaporated, and reconstituted in acetonitrile:0.1 mol/L ammonium hydroxide in water (1:1, vol/vol). The extracts were then analyzed using ultra-high performance liquid chromatography-tandem mass spectrometry. Validation experiments based on the ICH M10 guideline were carried out, and stability was evaluated under shipping and storage conditions. VAMS specimens were collected in the outpatient clinic to demonstrate the applicability of the assay.
RESULTS
The validated range of the method was considered accurate and precise for all analytes. Accordingly, the validation experiments met the relevant requirements, except for cross-analyte interference. Based on the stability data, shipment can be performed at room temperature within 14 days after sample collection and the VAMS specimen can be stored up to 9 months at -20 and -70°C. Samples from 59 patients were collected at the hospital.
CONCLUSIONS
The developed method could be used to successfully quantify the concentrations of abiraterone, D4A, alectinib, alectinib-M4, cabozantinib, imatinib, imatinib-M1, olaparib, sunitinib, and N-desethyl sunitinib within the validated range using VAMS. Therefore, the method can be used to estimate the dried whole blood-to-plasma ratios for TDM in the clinic.
PubMed: 38321598
DOI: 10.1097/FTD.0000000000001175 -
Journal of Pharmaceutical and... Apr 2024Serial blood sampling from one animal is useful to understand relationship between pharmacokinetics (PK) and pharmacological or toxicological events in individual...
Serial blood sampling from one animal is useful to understand relationship between pharmacokinetics (PK) and pharmacological or toxicological events in individual animals. To assess its feasibility in mice, two therapeutic antibodies were used to evaluate impacts by different blood sampling methods, sampling sites, and assay platforms on PK. Denosumab and Panitumumab were intravenously administered to mice and only 0.05 mL of blood sample per point was collected from jugular vein or tail vein. Blood samples were collected serially from a mouse or collected by traditional composite sampling from each mouse. Plasma concentrations of the two drugs were assayed by a generic ligand binding assay using Gyrolab or by a generic ultra-performance liquid chromatography with tandem mass spectrometry. The two assay platforms showed acceptable accuracy and precision and gave comparable PK parameters of the drugs, suggesting that both assays were successfully applied to the PK assessments. Comparable results in the PK profiles were noted between serial and composite blood samplings and differences in the two sampling sites did not impact PK. These findings suggest that microsampling combined with generic assays is useful to assess PK profiles of therapeutic antibodies in mice.
Topics: Mice; Animals; Tandem Mass Spectrometry; Chromatography, Liquid; Liquid Chromatography-Mass Spectrometry; Blood Specimen Collection; Dried Blood Spot Testing
PubMed: 38306865
DOI: 10.1016/j.jpba.2024.115993 -
Plant Methods Feb 2024Herbaria are becoming increasingly important as archives of biodiversity, and play a central role in taxonomic and biogeographic studies. There is also an ongoing...
BACKGROUND
Herbaria are becoming increasingly important as archives of biodiversity, and play a central role in taxonomic and biogeographic studies. There is also an ongoing interest in functional traits and the way they mediate interactions between a plant species and its environment. Herbarium specimens allow tracking trait values over time, and thus, capturing consequences of anthropogenic activities such as eutrophication. Here, we present an open, reproducible, non-destructive workflow to collect leaf trait data from herbarium specimens using near-infrared spectroscopy (NIRS), and a proof of concept for the reliability of this approach.
RESULTS
We carried out three experiments to test the suitability of non-destructive NIRS methods to predict leaf traits both for fresh and dried leaves: (1) With a fertilization experiment, we studied whether NIRS was able to capture changes in leaf N and leaf P during a fertilization experiment and we compared contents predicted by NIRS with results obtained from regular wet lab methods. Calibration models for leaf nitrogen and phosphorus contents had a quality of R = 0.7 and 0.5, respectively. We fitted calibration models for NIRS readings on fresh and dried leaf samples, both of which produced equally precise predictions compared to results from wet lab analyses. (2) We tested the effect of herbarium conservation on NIRS readings by simulating them through the application of six treatments combining freezing, drying and pesticide spraying in a factorial scheme and comparing these with untreated samples. No consistent changes were observed in the spectra quality before and after the simulated herbarium conditions. (3) Finally, we studied the effect of specimen storage duration using specimens from a 2018 study which were re-analyzed and compared with spectra obtained in 2021. No consistent changes in spectra were observed after the storage period.
CONCLUSIONS
The results demonstrate the reliability of NIRS to measure leaf N and P on herbarium samples. Together with the calibration method and dataset presented here, they provide a toolset allowing researchers to study the development of leaf traits and their response to environmental changes over decades and even centuries in a fast and non-destructive manner.
PubMed: 38303074
DOI: 10.1186/s13007-024-01146-x