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Journal of the American Society For... Jul 2024Over the past decades, proteomics has become increasingly important and a heavily discussed topic. The identification of intact proteins remains a major focus in this...
Over the past decades, proteomics has become increasingly important and a heavily discussed topic. The identification of intact proteins remains a major focus in this field. While most intact proteins are analyzed using high-resolution mass spectrometry, identifying them through low-resolution mass spectrometry continues to pose challenges. In our study, we investigated the capability of identifying various intact proteins using collision-induced dissociation (CID) and electron transfer without dissociation (ETnoD). Using myoglobin as our test protein, stable product ions were generated with CID, and the identities of the product ions were identified with ETnoD. ETnoD uses a short activation time (AcT, 5 ms) to create sequential charge-reduced precursor ion (CRI). The charges of the fragments and their sequences were determined with corresponding CRI. The product ions can be selected for subsequent CID (termed CID) combined with ETnoD for further sequence identification and validation. We refer to this method as CID/ETnoD. The use of a multistage CID activation (CID) and ETnoD protocol has been applied to several intact proteins to obtain multiple sequence identifications.
Topics: Myoglobin; Proteomics; Animals; Proteins; Amino Acid Sequence; Horses; Mass Spectrometry; Molecular Sequence Data; Tandem Mass Spectrometry
PubMed: 38905484
DOI: 10.1021/jasms.4c00108 -
Spectrochimica Acta. Part A, Molecular... Jun 2024Reaction between the polymeric [RuCl(CO)] and the N,N-bidentate ligand, 8-amino-quinoline (Quin), in methanol, afforded the photoactivated CO releasing molecule with the...
Reaction between the polymeric [RuCl(CO)] and the N,N-bidentate ligand, 8-amino-quinoline (Quin), in methanol, afforded the photoactivated CO releasing molecule with the formula of trans-(Cl,Cl)-[RuCl(CO)Quin]. In the presence of biomolecules or in solvents with varying polarity and coordinating abilities, the solvatochromic characteristics and dark stability were investigated. A new board band emerged in the visible spectrum during the illumination, and its position varies according to the type of solvent used, indicating the role of the solvent in controlling the nature of the CO-depleted species. Spectral methods were used in combination with density functional theory simulations to get insight into the local minimum structure and the electronic properties of the Ru(II) complex. The results of the myoglobin assay showed that within the first two hours of illumination, one of the two CO molecules was released. The cytotoxic properties of the Ru(II)-based complex were investigated against normal mice bone marrow stromal cells and malignant human acute monocytic leukaemia cells.
PubMed: 38901235
DOI: 10.1016/j.saa.2024.124644 -
BMC Sports Science, Medicine &... Jun 2024The study aimed to compare catecholamine, cortisol, and immune response in sprint- and endurance-trained athletes under the same training, aiming to observe if their...
BACKGROUND
The study aimed to compare catecholamine, cortisol, and immune response in sprint- and endurance-trained athletes under the same training, aiming to observe if their sport specialization affects these markers during a 9-day training camp.
METHODS
The study involved twenty-four young male (age 15.7 ± 1.6 years) and female (age 15.1 ± 1,3 years) athletes specializing in sprint and endurance athletics discipline. Blood samples for all measured parameters were taken at rested baseline, on the 4th day, and on the 9th day of training.
RESULTS
In both investigated groups a nonsignificant decrease in catecholamine levels was observed after 4 days of training, which remained stable throughout the camp. The cortisol level increased significantly in both athlete groups (sprint: T-0 vs. T-1 p = 0.0491; T-0 vs. T-3 p = 0.0001; endurance: T-0 vs. T-1 p = 0.0159; T-0 vs. T-3 p = 0.0005). The level of hs-CRP (sprint: T-0 vs. T-1 p = 0.0005; T-0 vs. T-3 p = 0.0001; endurance: T-0 vs. T-3 p = 0.0005), and myoglobin (sprint: T-0 vs. T-1 p = 0.0014; T-0 vs. T-3 p = 0.0001; endurance: T-0 vs. T-3 p = 0.0005) have increased and of hs-CRP and myoglobin level was significantly higher in sprint compared to endurance athletes (p < 0.05). The leukocyte level significantly decreased until the end of camp in both groups (sprint: T-0 vs. T-1 p = 0.0178; T-0 vs. T-3 p = 0.0175; endurance: T-0 vs. T-1 p = 0.0362; T-0 vs. T-3 p = 0.0362).
CONCLUSIONS
The applied training loads had a strong physiological impact leading to changes in stress hormones and immune responses depending on athletes` sport specialization. Training loads caused stronger responses in sprint athletes. However, both groups showed signs of severe fatigue development.
TRIAL REGISTRY
ClinicalTrials.gov ID: NCT06150105, retrospectively registered on 29.11.2023.
PubMed: 38898468
DOI: 10.1186/s13102-024-00924-3 -
Journal of Clinical Medicine May 2024It is unclear whether other cardiac biomarkers than NT-proBNP can be useful in the risk stratification of patients weaning from mechanical ventilation. The aim of this... (Review)
Review
It is unclear whether other cardiac biomarkers than NT-proBNP can be useful in the risk stratification of patients weaning from mechanical ventilation. The aim of this study is to summarize the role of ischemic cardiac biomarkers in predicting spontaneous breathing trial (SBT) or extubation failure. : We systematically searched Embase, MEDLINE, Web of Science, and Cochrane Central for studies published before January 2024 that reported the association between ischemic cardiac biomarkers and SBT or extubation failure. Data were extracted using a standardized form and methodological assessment was performed using the QUIPS tool. Seven observational studies investigating four ischemic cardiac biomarkers (Troponin-T, Troponin-I, CK-MB, Myoglobin) were included. One study reported a higher peak Troponin-I in patients with extubation failure compared to extubation success (50 ng/L [IQR, 20-215] versus 30 ng/L [IQR, 10-86], = 0.01). A second study found that Troponin-I measured before the SBT was higher in patients with SBT failure in comparison to patients with SBT success (100 ± 80 ng/L versus 70 ± 130 ng/L, = 0.03). A third study reported a higher CK-MB measured at the end of the SBT in patients with weaning failure (SBT or extubation failure) in comparison to weaning success (8.77 ± 20.5 ng/mL versus 1.52 ± 1.42 ng/mL, = 0.047). Troponin-T and Myoglobin as well as Troponin-I and CK-MB measured at other time points were not found to be related to SBT or extubation failure. However, most studies were underpowered and with high risk of bias. : The association with SBT or extubation failure is limited for Troponin-I and CK-MB and appears absent for Troponin-T and Myoglobin, but available studies are hampered by significant methodological drawbacks. To more definitively determine the role of ischemic cardiac biomarkers, future studies should prioritize larger sample sizes, including patients at risk of cardiac disease, using stringent SBTs and structured timing of laboratory measurements before and after SBT.
PubMed: 38892952
DOI: 10.3390/jcm13113242 -
Food Chemistry Jun 2024This study compared the proteomics of beef patties under high‑oxygen modified atmosphere packaging (HiOx-MAP) and vacuum packaging (VP) during heating. The color and...
This study compared the proteomics of beef patties under high‑oxygen modified atmosphere packaging (HiOx-MAP) and vacuum packaging (VP) during heating. The color and oxidation stability of fresh patties, and myoglobin denaturation of cooked patties were also measured. The results suggested that HiOx-MAP patties contained more oxymyoglobin in fresh meat and had higher myoglobin denaturation during heating than VP patties, resulting in premature browning (PMB) during cooking. Proteomic analysis found that the overabundance of proteasome subunit beta type-2 (PSMB2) and peroxiredoxin-2 (PRDX2) in HiOx-55 °C, which can remove the damaged proteins and inhibit oxidation respectively, are of benefit to meat color stability during storage, however, this was still insufficient to inhibit the occurrence of PMB during cooking. The high abundance of lamin B1 (LMNB1) in VP-55 °C can maintain the stability of meat color. This research provides greater understanding, based on proteomic perspectives, of the molecular mechanism of PMB.
PubMed: 38876067
DOI: 10.1016/j.foodchem.2024.140022 -
Journal of Applied Physiology... Jun 2024Prior studies of muscle blood flow and muscle specific oxygen consumption have required invasive injection of dye and Magnetic Resonance Imaging, respectively. Such...
Prior studies of muscle blood flow and muscle specific oxygen consumption have required invasive injection of dye and Magnetic Resonance Imaging, respectively. Such measures have limited utility for continuous monitoring of the respiratory muscles. Frequency domain near-infrared spectroscopy and diffuse correlation spectroscopy (FD-NIRS & DCS) can provide continuous surrogate measures of blood flow index (BFi) and metabolic rate of oxygen consumption (MRO). This study aimed to validate sternocleidomastoid FD-NIRS & DCS outcomes against electromyography (EMG) and mouth pressure (Pm) during incremental inspiratory threshold loading (ITL). Six females and six male healthy adults (mean±SD; 30±7 years, maximum inspiratory pressure 118±61 cmHO) performed incremental ITL starting at low loads (8±2 cmHO) followed by 50g increments every two minutes until task failure. FD-NIRS & DCS continuously measured sternocleidomastoid oxygenated and deoxygenated hemoglobin+myoglobin (oxy/deoxy[Hb+Mb]), tissue saturation of oxygen (StO), BF, and MRO. Ventilatory parameters including inspiratory Pm were also evaluated. Pm increased during incremental ITL (<0.05), reaching -47[-74 - -34] cmHO (median[25%-75%IQR] at task failure. Ventilatory parameters were constant throughout ITL (all >0.05). Sternocleidomastoid BFi and MRO increased from the start of the ITL (both P<0.05). Deoxy[Hb+Mb] increased close to task failure, concomitantly with a constant increase in MRO, and decreased StO. Sternocleidomastoid deoxy[Hb+Mb], BF, StO and MRO obtained during ITL via FD-NIRS & DCS correlated with sternocleidomastoid EMG (all <0.05). In healthy adults, FD-NIRS & DCS can provide continuous surrogate measures of respiratory BFi and MRO-. Increasing sternocleidomastoid oxygen consumption near task failure was associated with increased oxygen extraction and reduced tissue saturation.
PubMed: 38867669
DOI: 10.1152/japplphysiol.00871.2023 -
Revista Da Associacao Medica Brasileira... 2024Chemotherapy with doxorubicin may lead to left ventricular dysfunction. There is a controversial recommendation that biomarkers can predict ventricular dysfunction,... (Observational Study)
Observational Study
BACKGROUND
Chemotherapy with doxorubicin may lead to left ventricular dysfunction. There is a controversial recommendation that biomarkers can predict ventricular dysfunction, which is one of the most feared manifestations of anthracycline cardiotoxicity.
OBJECTIVE
The aim of this study was to evaluate the behavior of biomarkers such as Troponin I, type B natriuretic peptide, creatine phosphokinase fraction MB, and myoglobin in predicting cardiotoxicity in a cohort of women with breast cancer undergoing chemotherapy with anthracycline.
METHODS
This is an observational, prospective, longitudinal, unicentric study, which included 40 women with breast cancer, whose therapeutic proposal included treatment with doxorubicin. The protocol had a clinical follow-up of 12 months. Biomarkers such as Troponin I, type B natriuretic peptide, creatine phosphokinase fraction MB, and myoglobin were measured pre-chemotherapy and after the first, third, fourth, and sixth cycles of chemotherapy.
RESULTS
There was a progressive increase in type B natriuretic peptide and myoglobin values in all chemotherapy cycles. Although creatine phosphokinase fraction MB showed a sustained increase, this increase was not statistically significant. Troponin, type B natriuretic peptide, myoglobin, and creatine phosphokinase fraction MB were the cardiotoxicity markers with the earliest changes, with a significant increase after the first chemotherapy session. However, they were not able to predict cardiotoxicity.
CONCLUSION
Troponin I, type B natriuretic peptide, myoglobin, and creatine phosphokinase fraction MB are elevated during chemotherapy with doxorubicin, but they were not able to predict cardiotoxicity according to established clinical and echocardiographic criteria. The incidence of subclinical cardiotoxicity resulting from the administration of doxorubicin was 12.5%.
Topics: Humans; Female; Breast Neoplasms; Prospective Studies; Troponin I; Doxorubicin; Cardiotoxicity; Middle Aged; Biomarkers; Myoglobin; Adult; Antibiotics, Antineoplastic; Natriuretic Peptide, Brain; Aged; Creatine Kinase, MB Form; Longitudinal Studies; Anthracyclines; Ventricular Dysfunction, Left; Predictive Value of Tests
PubMed: 38865526
DOI: 10.1590/1806-9282.2024S106 -
ACS Applied Materials & Interfaces Jun 2024This paper reports a microfluidic device for the electrochemical and plasmonic detection of cardiac myoglobin (cMb) and cardiac troponin I (cTnI) with noticeable limits...
This paper reports a microfluidic device for the electrochemical and plasmonic detection of cardiac myoglobin (cMb) and cardiac troponin I (cTnI) with noticeable limits of detection (LoD) as low as a few picograms per milliliter (pg/mL) ranges, achieved in a short detection time. The device features two working electrodes, each with a mesoporous NiVO nanoscaffold grafted with reduced graphene oxide (rGO) that improves the interaction of diffusing analyte molecules with the sensing surface by providing a high surface area and reaction kinetics. Electrochemical studies reveal sensitivities as high as 9.68 μA ng/mL and a LoD of 2.0 pg/mL for cTnI, and 8.98 μA ng/mL and 4.7 pg/mL for cMb. Additionally, the surface plasmon resonance (SPR) studies demonstrate a low-level LoD of 8.8 pg/mL for cMb and 7.3 pg/mL for cTnI. The dual-modality sensor enables dynamic tracking of kinetic antigen-antibody interactions during sensing, self-verification through providing signals of two modes, and reduced false readout. This study demonstrates the complementary nature of the electrochemical and SPR modes in biosensing, with the electrochemical mode being highly sensitive and the SPR mode providing superior tracking of molecular recognition behaviors. The presented sensor represents a significant innovation in cardiovascular disease management and can be applied to monitor other clinically important biomolecules.
Topics: Myocardial Infarction; Troponin I; Graphite; Electrochemical Techniques; Myoglobin; Surface Plasmon Resonance; Humans; Porosity; Biosensing Techniques; Limit of Detection; Lab-On-A-Chip Devices; Nanostructures
PubMed: 38860871
DOI: 10.1021/acsami.4c01398 -
Journal of Inorganic Biochemistry Sep 2024The NO dioxygenation reaction catalyzed by heme-containing globin proteins is a crucial aerobic detoxification pathway. Accordingly, the second order reaction of NO with...
The NO dioxygenation reaction catalyzed by heme-containing globin proteins is a crucial aerobic detoxification pathway. Accordingly, the second order reaction of NO with oxymyoglobin and oxyhemoglobin has been the focus of a large number of kinetic and spectroscopic studies. Stopped-flow and rapid-freeze-quench (RFQ) measurements have provided evidence for the formation of a Fe(III)-nitrato complex with millisecond lifetime prior to release of the nitrate product, but the temporal resolution of these techniques is insufficient for the characterization of precursor species. Most mechanistic models assume the formation of an initial Fe(III)-peroxynitrite species prior to homolytic cleavage of the OO bond and recombination of the resulting NO and Fe(IV)=O species. Here we report vibrational spectroscopy measurements for the reaction of oxymyoglobin with a photolabile caged NO donor at cryogenic temperatures. We show that this approach offers efficient formation and trapping of the Fe(III)-nitrato, enzyme-product, complex at 180 K. Resonance Raman spectra of the Fe(III)-nitrato complex trapped via RFQ in the liquid phase and photolabile NO release at cryogenic temperatures are indistinguishable, demonstrating the complementarity of these approaches. Caged NO is released by irradiation <180 K but diffusion into the heme pocket is fully inhibited. Our data provide no evidence for Fe(III)-peroxynitrite of Fe(IV)=O species, supporting low activation energies for the NO to nitrate conversion at the oxymyoglobin reaction site. Photorelease of NO at cryogenic temperatures allows monitoring of the reaction by transmittance FTIR which provides valuable quantitative information and promising prospects for the detection of protein sidechain reorganization events in NO-reacting metalloenzymes.
Topics: Myoglobin; Nitric Oxide; Spectrum Analysis, Raman; Cold Temperature; Animals; Kinetics; Vibration
PubMed: 38852292
DOI: 10.1016/j.jinorgbio.2024.112633 -
Structure (London, England : 1993) Jun 2024In a recent issue of Nature, Barends et al. studied the photodissociation of carboxymyoglobin with ultrafast laser pump-probe serial femtosecond crystallography...
In a recent issue of Nature, Barends et al. studied the photodissociation of carboxymyoglobin with ultrafast laser pump-probe serial femtosecond crystallography experiments. They observed significant differences in heme protein structural dynamics for biologically relevant 1-photon excitation relative to high excitation leading to the absorption of several photons per heme.
Topics: Myoglobin; Protein Conformation; Heme; Crystallography, X-Ray; Light; Photons; Models, Molecular
PubMed: 38848681
DOI: 10.1016/j.str.2024.05.008