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1.
Plant J ; 2024 Jul 03.
Artículo en Inglés | MEDLINE | ID: mdl-38961633

RESUMEN

Global wheat production amounted to >780 MMT during 2022-2023 whose market size are valued at >$128 billion. Wheat is highly susceptible to high-temperature stress (HTS) throughout the life cycle and its yield declines 5-7% with the rise in each degree of temperature. Previously, we reported an array of HTS-response markers from a resilient wheat cv. Unnat Halna and described their putative role in heat acclimation. To complement our previous results and identify the key determinants of thermotolerance, here we examined the cytoplasmic proteome of a sensitive cv. PBW343. The HTS-triggered metabolite reprograming highlighted how proteostasis defects influence the formation of an integrated stress-adaptive response. The proteomic analysis identified several promising HTS-responsive proteins, including a NACα18 protein, designated TaNACα18, whose role in thermotolerance remains unknown. Dual localization of TaNACα18 suggests its crucial functions in the cytoplasm and nucleus. The homodimerization of TaNACα18 anticipated its function as a transcriptional coactivator. The complementation of TaNACα18 in yeast and overexpression in wheat demonstrated its role in thermotolerance across the kingdom. Altogether, our results suggest that TaNACα18 imparts tolerance through tight regulation of gene expression, cell wall remodeling and activation of cell defense responses.

2.
Bioorg Med Chem Lett ; 108: 129789, 2024 Aug 01.
Artículo en Inglés | MEDLINE | ID: mdl-38729318

RESUMEN

Receptors are proteinous macromolecules which remain in the apo form under normal/unliganded conditions. As the ligand approaches, there are specific stereo-chemical changes in the apo form of the receptor as per the stereochemistry of a ligand. Accordingly, a series of substituted dimethyl-chroman-based stereochemically flexible and constrained Tamoxifen analogs were synthesized as anti-breast cancer agents. The synthesized compounds 19a-e, 20a-e, 21, and 22a-e, showed significant antiproliferative activity against estrogen receptor-positive (ER+, MCF-7) and negative (ER-, MDA MB-231) cells within IC50 value 8.5-25.0 µM. Amongst all, four potential molecules viz 19b, 19e, 22a, and 22c, were evaluated for their effect on the cell division cycle and apoptosis of ER+ and ER- cancer cells (MCF-7 & MDA MB-231cells), which showed that these compounds possessed antiproliferative activity through triggering apoptosis. In-silico docking experiments elucidated the possible affinity of compounds with estrogen receptors-α and -ß.


Asunto(s)
Antineoplásicos , Apoptosis , Neoplasias de la Mama , Proliferación Celular , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Antineoplásicos/farmacología , Antineoplásicos/síntesis química , Antineoplásicos/química , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/patología , Proliferación Celular/efectos de los fármacos , Estereoisomerismo , Relación Estructura-Actividad , Línea Celular Tumoral , Apoptosis/efectos de los fármacos , Cromanos/farmacología , Cromanos/síntesis química , Cromanos/química , Simulación del Acoplamiento Molecular , Receptor alfa de Estrógeno/metabolismo , Receptor alfa de Estrógeno/antagonistas & inhibidores , Femenino , Estructura Molecular , Células MCF-7 , Relación Dosis-Respuesta a Droga , Tamoxifeno/farmacología , Tamoxifeno/síntesis química , Tamoxifeno/química
3.
Cell Physiol Biochem ; 57(6): 452-477, 2023 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-37978922

RESUMEN

BACKGROUND/AIMS: All body functions are activated, synchronized and controlled by a substantial, complex network, the nervous system. Upon injury, pathophysiology of the nerve injury proceeds through different paths. The axon may undergo a degenerative retraction from the site of injury for a short distance unless the injury is near to the cell body, in which case it continues to the soma and undergoes retrograde neuronal degeneration. Otherwise, the distal section suffers from Wallerian degeneration, which is marked by axonal swelling, spheroids, and cytoskeleton degeneration. The objective of the study was to evaluate the potential of mesenchymal stem cell laden neural scaffold and insulin-like growth factor I (IGF-I) in nerve regeneration following sciatic nerve injury in a rat model. METHODS: The animals were anaesthetized and a cranio-lateral incision over left thigh was made. Sciatic nerve was exposed and crush injury was introduced for 90 seconds using haemostat at second locking position. The muscle and skin were sutured in routine fashion and thus the rat model of sciatic crush injury was prepared. The animal models were equally distributed into 5 different groups namely A, B, C, D and E and treated with phosphate buffer saline (PBS), carbon nanotubes based neural scaffold only, scaffold with IGF-I, stem cell laden scaffold and stem cell laden scaffold with IGF-I respectively. In vitro scaffold testing was performed. The nerve regeneration was assessed based on physico-neuronal, biochemical, histopathological examination, and relative expression of NRP-1, NRP-2 and GAP-43 and scanning electron microscopy. RESULTS: Sciatic nerve injury model with crush injury produced for 90 seconds was standardized and successfully used in this study. All the biochemical parameters were in normal range in all the groups indicating no scaffold related changes. Physico-neuronal, histopathological, relative gene expression and scanning electron microscopy observations revealed appreciable nerve regeneration in groups E and D, followed by C and B. Restricted to no regeneration was observed in group A. CONCLUSION: Carbon nanotubes based scaffold provided electro-conductivity for proper neuronal regeneration while rat bone marrow-derived mesenchymal stem cells were found to induce axonal sprouting, cellular transformation; whereas IGF-I induced stem cell differentiation, myelin synthesis, angiogenesis and muscle differentiation.


Asunto(s)
Lesiones por Aplastamiento , Células Madre Mesenquimatosas , Nanotubos de Carbono , Neuropatía Ciática , Ratas , Animales , Ratas Wistar , Factor I del Crecimiento Similar a la Insulina/farmacología , Factor I del Crecimiento Similar a la Insulina/uso terapéutico , Neuropatía Ciática/tratamiento farmacológico , Neuropatía Ciática/patología , Nervio Ciático/lesiones , Regeneración Nerviosa/fisiología , Lesiones por Aplastamiento/tratamiento farmacológico , Lesiones por Aplastamiento/patología , Células Madre Mesenquimatosas/patología , Colágeno
4.
Physiol Plant ; 175(2): e13895, 2023 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-37096431

RESUMEN

Everyone likes to add color to their life, and plants use them too. However, unlike humans, plants contain natural pigments to add color to fruits, leaves, and vegetables. Plants produce a variety of phytopigments, such as flavonoids, carotenoids, and anthocyanins, which play essential roles in plant stress tolerance. Leveraging natural phytopigments to develop stress-resilient crops will require an in-depth understanding of pigment formation and function. In this context, Zhang et al. (2023) studied the role of MYB6 and bHLH111 in enhancing anthocyanin biosynthesis in petals during drought.


Asunto(s)
Antocianinas , Flores , Humanos , Plantas , Flavonoides , Hojas de la Planta , Regulación de la Expresión Génica de las Plantas
5.
J Fluoresc ; 33(5): 1961-1970, 2023 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-36930343

RESUMEN

Herein, we designed two fluorescent tetrasubstituted benzene sulfonyl appended Thiacalix[4]arene receptors named L1 and L2, which sensitively and selectively detect Sulfosulfuron among other herbicides and pesticides. The detection limit (LOD) was found to be 0.21 ppm and 0.35 ppm, and the enhancement constant (Ks) was determined to be 7.07 X 104 M-1 and 5.55 X 104 M-1 for L1 and L2, respectively. Using the non-linear regression method, the association constant was obtained as 2.1 X 104 M-1 and 2.23 X 104 M-1 whereas, the binding ratio was found to be 1:1 for both L1 and L2, respectively. Additionally, the interference studies show the selective nature of receptors for Sulfosulfuron among its sulfonylurea family. To further confirm the interaction mechanism, 1H-NMR spectroscopy, and a computational investigation were carried out, which validates the 1:1 binding ratio. The receptors were found to be recyclable in nature with simple acid-base treatment. This new approach of using supramolecules as fluorescent probes for sensitive and selective detection of herbicides is rare in the literature.

6.
Plant J ; 105(5): 1374-1389, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-33283912

RESUMEN

The molecular mechanism of high-temperature stress (HTS) response, in plants, has so far been investigated using transcriptomics, while the dynamics of HTS-responsive proteome remain unexplored. We examined the adaptive responses of the resilient wheat cultivar 'Unnat Halna' and dissected the HTS-responsive proteome landscape. This led to the identification of 55 HTS-responsive proteins (HRPs), which are predominantly involved in metabolism and defense pathways. Interestingly, HRPs included a 2-cysteine peroxiredoxin (2CP), designated Ta2CP, presumably involved in stress perception and adaptation. Complementation of Ta2CP in yeast and heterologous expression in Arabidopsis demonstrated its role in thermotolerance. Both Ta2CP silencing and overexpression inferred the involvement of Ta2CP in plant growth and chlorophyll biosynthesis. We demonstrated that Ta2CP interacts with protochlorophyllide reductase b, TaPORB. Reduced TaPORB expression was found in Ta2cp-silenced plants, while upregulation was observed in Ta2CP-overexpressed plants. Furthermore, the downregulation of Ta2CP in Taporb-silenced plants and reduction of protochlorophyllide in Ta2cp-silenced plants suggested the key role of Ta2CP in chlorophyll metabolism. Additionally, the transcript levels of AGPase1 and starch were increased in Ta2cp-silenced plants. More significantly, HTS-treated Ta2cp-silenced plants showed adaptive responses despite increased reactive oxygen species and peroxide concentrations, which might help in rapid induction of high-temperature acclimation.


Asunto(s)
Peroxirredoxinas/metabolismo , Proteínas de Plantas/metabolismo , Triticum/metabolismo , Antioxidantes/metabolismo , Citoplasma/metabolismo , Regulación de la Expresión Génica de las Plantas , Calor , Proteínas de Plantas/genética , Triticum/genética
7.
J Fluoresc ; 32(2): 583-592, 2022 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-35013852

RESUMEN

Identification and detection of harmful contaminants such as nickel and other materials from soil and water is critical necessity at the present moment. So with this motive to detect and identify harmful pollutants, a novel cyclotriveratrylene based derivative was prepared for the detection and binding of harmful pollutants which had the properties of fluorescence. The newly derivative of Cyclotriveratrylene was found to be highly sensitive and selective towards Ni2+ ions. The complexation behaviour of this newly synthesised molecule was studied in presence of transition elements. Also computational methods such as docking, molecular modelling and DFT were used to study the molecular orbitals and energies of CTG-NBEP. The detection of Ni2+ from water samples were also carried out successfully.


Asunto(s)
Técnicas Biosensibles , Níquel/análisis , Compuestos Policíclicos/química , Contaminantes del Suelo/análisis , Espectrometría de Fluorescencia/métodos , Contaminantes del Agua/análisis , Teoría Funcional de la Densidad , Iones/análisis , Simulación del Acoplamiento Molecular
8.
J Fluoresc ; 32(1): 215-226, 2022 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-34694547

RESUMEN

A new and innovative fluorescent structure was constructed on Cyclotriveratrylene affiliated to Dansyl chloride (DNSC) and was used to detect Cr (III) and Fe (III) among the various cations by using spectrofluorimetric method. The characterization of the new compound was carried out using the 1H-NMR, 13C-NMR, and ESI-MS techniques. The interaction and role of DNSC-CTV with cations was reviewed. A change in the spectra of absorption directed to the conclusion that there is substantial interaction of Cr (III) and Fe (III) with DNSC-CTV. Furthermore the interaction of the ligand DNSC-CTV with the metal ions Chromium (III) and Iron (III) showed quenching in the emission spectra. Quantum yield of the complexes were calculated and the stern volmer analysis was done to deduce the quenching mechanism of fluorescence to being either static or dynamic. The molecule DNSC-CTV was further studied with the help of computational methods such as molecular docking to study the binding interactions and properties of the molecule.


Asunto(s)
Técnicas Biosensibles/métodos , Cromo/análisis , Colorantes Fluorescentes , Fluorometría/métodos , Hierro/análisis , Espectrometría de Fluorescencia/métodos , Cationes , Compuestos de Dansilo/química , Simulación del Acoplamiento Molecular , Compuestos Policíclicos/química
9.
J Fluoresc ; 32(4): 1279-1288, 2022 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-35543793

RESUMEN

A new tri-naphthoylated Cyclotriveratrylene molecule has been synthesized for the rapid and sensitive detection of 4-nitrotoluene (4-NT) among various nitro aromatic compounds (NACs) by using a spectrofluorimetric method. The newly derrivatized cyclotriveratrylene compound is successfully confirmed by using the available techniques of 1H-NMR, 13CNMR, and ESI-MS. This synthesised molecule is 1NC-CTV (1-Naptholy Chloride-Cyclotriveratrylene). Strong quenching in the fluorescence intensity of 1NC-CTV was observed upon the addition of 4-NT. Further quantum yield studies were carried out and by using the stern volmer it was concluded that the fluorescence quenching mechanism is dynamic or static. The molecule 1NC-CTV was further studied with the help of computational methods such as molecular docking to study the binding interactions and properties of the molecule.


Asunto(s)
Tolueno , Modelos Moleculares , Simulación del Acoplamiento Molecular , Espectrometría de Fluorescencia , Tolueno/química
10.
J Med Virol ; 93(4): 2406-2419, 2021 04.
Artículo en Inglés | MEDLINE | ID: mdl-33347622

RESUMEN

The analyses of 2325 severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) genomes revealed 107, 162, and 65 nucleotide substitutions in the coding region of SARS-CoV-2 from the three continents America, Europe, and Asia, respectively. Of these nucleotide substitutions 58, 94, and 37 were nonsynonymous types mostly present in the Nsp2, Nsp3, Spike, and ORF9. A continent-specific phylogram analyses clustered the SARS-CoV-2 in the different group based on the frequency of nucleotide substitutions. Detailed analyses about the continent-specific amino acid changes and their effectiveness by SNAP2 software was investigated. We found 11 common nonsynonymous mutations; among them, two novel effective mutations were identified in ORF9 (S194L and S202N). Intriguingly, ORF9 encodes nucleocapsid phosphoprotein possessing many effective mutations across continents and could be a potential candidate after the spike protein for studying the role of mutation in viral assembly and pathogenesis. Among the two forms of certain frequent mutation, one form is more prevalent in Europe continents (Nsp12:L314, Nsp13:P504, Nsp13:Y541, Spike:G614, and ORF8:L84) while other forms are more prevalent in American (Nsp12:P314, Nsp13:L504, Nsp13:C541, Spike:D614, and ORF8:L84) and Asian continents (Spike:D614), indicating the spatial and temporal dynamics of SARS-CoV-2. We identified highly conserved 38 regions and among these regions, 11 siRNAs were predicted on stringent criteria that can be used to suppress the expression of viral genes and the corresponding reduction of human viral infections. The present investigation provides information on different mutations and will pave the way for differentiating strains based on virulence and their use in the development of better antiviral therapy.


Asunto(s)
COVID-19/virología , Mutación , SARS-CoV-2/genética , Antivirales/farmacología , Asia/epidemiología , COVID-19/epidemiología , Proteínas de la Nucleocápside de Coronavirus/genética , Proteasas Similares a la Papaína de Coronavirus/genética , Europa (Continente)/epidemiología , Silenciador del Gen , Genes Virales , Genoma Viral , Humanos , Sistemas de Lectura Abierta , Fosfoproteínas/genética , Filogenia , ARN Interferente Pequeño/genética , SARS-CoV-2/clasificación , SARS-CoV-2/efectos de los fármacos , Proteínas no Estructurales Virales/genética , Proteínas Virales/genética , Tratamiento Farmacológico de COVID-19
11.
Eur J Pediatr ; 180(3): 767-773, 2021 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-32813124

RESUMEN

Shielding the precordium can effect manifestation of haemodynamically significant patent ductus arteriosus (hsPDA). Preterm neonates born at ≤ 32 weeks of gestation if needed phototherapy within 72 h of birth and had no echocardiographically proven hsPDA were eligible to be enrolled in this open-label randomised controlled trial. In chest shielding group, in addition to the standard care, left side of the chest was covered using food grade aluminium foil during phototherapy while control group received standard care. Mean gestational age (weeks; 30.1 ± 1.5 vs 30.1 ± 1.6) was comparable in the two groups. However, neonates in the chest shield group had lower birth weight (g; 1281 ± 259 vs 1422 ± 307) and were more likely to be small-for-gestational age (21.6% vs 8.0%). It was seen that 4 (7.8%) babies in the chest shield group and 5 (10%) babies in the standard group developed hsPDA after starting phototherapy with relative risk (RR) of 0.78 (95% CI 0.22-2.75). The left atrium to aortic ratio was significantly different in the two groups with 1.5 ± 0.1 in the chest shield group and 1.8 ± 0.2 in standard group (p value 0.03).Conclusion: Chest shielding of preterm babies during phototherapy has no effect on the incidence of haemodynamically significant patent ductus arteriosus.Trial registration: Trial was registered with Clinical trial registry of India (CTRI/2018/01/011069). What is Known: • Chest shielding in preterm neonates under phototherapy has inconclusive effect on the manifestation of patent ductus arteriosus. What is New: • Preterm neonates under phototherapy have no significant difference in manifestation of haemodynamically significant patent ductus arteriosus if precordium is shielded.


Asunto(s)
Conducto Arterioso Permeable , Recien Nacido Prematuro , Conducto Arterioso Permeable/terapia , Humanos , India , Recién Nacido de Bajo Peso , Recién Nacido , Fototerapia
12.
PLoS Med ; 17(3): e1003087, 2020 03.
Artículo en Inglés | MEDLINE | ID: mdl-32218564

RESUMEN

BACKGROUND: In settings of mass displacement, unaccompanied minors (UAMs) are recognized as a vulnerable group and consequently prioritized by relief efforts. This study examines how the interpretation of vulnerability by the national shelter system for male UAMs in Greece shapes their trajectories into adulthood. METHODS AND FINDINGS: Between August 2018 and April 2019, key informant interviews were carried out with child protection staff from Greek non-governmental organizations that refer UAMs to specialized children's shelters in Athens to understand how child protection workers interpret vulnerability. In-depth interviews and life history calendars were collected from 44 male migrant youths from Afghanistan, Pakistan, Bangladesh, and Iran who arrived in Greece as UAMs but had since transitioned into adulthood. Analysis of in-depth interviews and life history calendars examined how cumulative disadvantage and engagement with the shelter system altered youths' trajectories into adulthood. Younger adolescents were perceived as more vulnerable and prioritized for shelters over those who were "almost 18" years old. However, a subset of youths who requested shelter at the age of 17 years had experienced prolonged journeys where they spent months or years living on their own in socially isolated environments that excluded them from experiences conducive to adolescent development. The shelter system for UAMs in Greece enabled youths to develop new skills and networks that facilitated integration into society, and transferred them into adult housing when they turned 18 years old so that they could continue developing new skills. Those who were not in shelters by age 18 years could not access adult housing and lost this opportunity. Limitations included possible underrepresentation of homeless youth as well as the inability to capture all nationalities of UAMs in Greece, though the 2 most common nationalities, Afghan and Pakistani, were included. CONCLUSIONS: Due to the way vulnerability was interpreted by the shelter system for UAMs, youths who had the greatest need to learn new skills to facilitate their integration often had the least opportunity to do so. To avoid creating long-lasting disparities between UAMs who are placed in shelters and those who are not, pathways should be developed to allow young adult males to enter accommodation facilities and build skills and networks that facilitate integration. Furthermore, cumulative disadvantages should be taken into account while assessing UAMs' vulnerability. Following UAMs' trajectories into early adulthood was critical for capturing this long-term consequence of the shelter system's interpretation of vulnerability.


Asunto(s)
Conducta del Adolescente , Servicios de Protección Infantil , Jóvenes sin Hogar/psicología , Vivienda , Inmigrantes Indocumentados/psicología , Poblaciones Vulnerables/psicología , Actividades Cotidianas , Adaptación Psicológica , Adolescente , Factores de Edad , Emigración e Inmigración , Grecia , Humanos , Entrevistas como Asunto , Masculino , Investigación Cualitativa , Conducta Social , Adulto Joven
14.
Plant Physiol ; 2024 May 10.
Artículo en Inglés | MEDLINE | ID: mdl-38728436
15.
Am J Orthod Dentofacial Orthop ; 158(5): 684-693, 2020 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-33010980

RESUMEN

INTRODUCTION: This retrospective pilot study assessed the pre- and posttreatment lip profile changes among adult patients with Class II Division 1 malocclusion of varied growth patterns and compared these changes with patients with a skeletal Class I relationship, to identify the dental, skeletal, and soft tissue cephalometric variables that altered the posttreatment lip profile. METHODS: A total of 33 digital cephalograms were divided into 4 study groups based on the mandibular plane (Mp-SN) angle: group 1 (control, skeletal Class I; mean angle = 21.5°) (n = 8), group 2 (Class II Division 1 malocclusion; low angle = <26°) (n = 8), group 3 (Class II Division 1 malocclusion; intermediate angle = 26°-38°) (n = 9), and group 4 (Class II Division 1 malocclusion; high angle = >38°) (n = 8). RESULTS: We observed a significant decrease in SNA, ANB, maxillary incisor to NA (degrees and millimeters), and soft tissue parameters, especially lip strain and lower lip to E line and upper lip to E line. Posttreatment intergroup comparison showed a significant increase of Mp-SN, and this increase was greater in group 3, followed by group 4 as compared with group 1. In addition, statistically significant differences in SNA, ANB, mandibular incisor to NB (degrees) and IMPA, and H angle in groups 2-4 were observed. CONCLUSIONS: Patients with Class II Division 1 malocclusion showed a significant decrease in SNA, ANB, maxillary incisor to NA, and all soft tissue parameters in posttreatment compared with group 1. Thus, the soft tissue changes resulted in a similar profile to patients in group 1, who are considered to have an aesthetically pleasing posttreatment profile.


Asunto(s)
Labio , Maloclusión Clase II de Angle , Adulto , Diente Premolar/cirugía , Cefalometría , Humanos , Labio/anatomía & histología , Labio/diagnóstico por imagen , Maloclusión Clase II de Angle/diagnóstico por imagen , Maloclusión Clase II de Angle/terapia , Proyectos Piloto , Estudios Retrospectivos
18.
Plant Physiol ; 192(4): 2596-2597, 2023 08 03.
Artículo en Inglés | MEDLINE | ID: mdl-37072322
19.
Plant Physiol ; 2023 Dec 14.
Artículo en Inglés | MEDLINE | ID: mdl-38096243
20.
Rapid Commun Mass Spectrom ; 33(2): 203-214, 2019 Jan 30.
Artículo en Inglés | MEDLINE | ID: mdl-30304582

RESUMEN

RATIONALE: Acid digestion of carbonates to release CO2 is a crucial and sensitive step in sample preparation for clumped isotope analysis. In addition to data reduction and instrumental artefacts, many other uncertainties in the clumped isotope analysis of carbonates arise from the method used for the preparation of CO2 . We describe here an in-house-designed reaction vessel that circumvents degassing and contamination problems commonly associated with the McCrea-type digestion protocols. METHODS: We designed a leak-free break seal reaction vessel (made of Pyrex™) suitable for prolonged acid digestion at 25°C. Using this new vessel, several carbonate reference materials widely used in the clumped isotope community and other in-house laboratory standards were acid-digested and analysed for their δ13 C, δ18 O and Δ47 values with a dual inlet MAT 253 isotope ratio mass spectrometer following standard gas chromatography purification and data evaluation protocols. RESULTS: Long-term reproducibility in Δ47 determination was established using international references and in-house working standards as follows (mean and SE): Carrara-1 (0.395 ± 0.002‰, n = 43), Carrara-2 (0.441 ± 0.003‰, n = 22), OMC (0.587 ± 0.004‰, n = 16), NBS 19 (0.393 ± 0.005‰, n = 10), NBS 18 (0.473 ± 0.003‰, n = 5), ETH 1 (0.271 ± 0.005‰, n = 7), ETH 3 (0.698 ± 0.005‰, n = 3), MZ (0.715 ± 0.002‰, n = 3) and several others. CONCLUSIONS: A new method using a break seal tube was found to be efficient for the clumped isotope analysis of carbonates that require longer reaction time at 25°C. This method yields good precision in Δ47 analysis and was found to be suitable for acid digestions at any desired temperature.

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