Micro-Raman Spectroscopy Analysis of Optically Trapped Erythrocytes in Jaundice

Jacob, S.; Bankapur, Aseefhali; Barkur, Surekha; Acharya, Mahendra; Chidangil, Santhosh; Rao, P. L. N. G.; Kamath, Asha; Lakshmi, R.V.; Baby, Prathap Moothamadathil; Rao, Raghavendra K.

doi:10.3389/fphys.2020.00821

Cited by 5 publications

(9 citation statements)

References 41 publications

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“…Following TM treatment, the color of the liver was yellowish, implying the presence of jaundice. Bilirubin is a vital basis for the clinical diagnosis of jaundice because dysfunction of the hepatobiliary system leads to an unhealthy buildup of bilirubin in the blood, resulting in jaundice ( Jacob et al, 2020 ). In this study, TBIL increased significantly in both the LD and HD groups, indicating the occurrence of jaundice, and DBIL and GGT levels increased significantly in the HD group, illustrating the occurrence of liver jaundice.…”

Section: Discussionmentioning

confidence: 99%

Acute Endoplasmic Reticulum Stress Induces Inflammation Reaction, Complement System Activation, and Lipid Metabolism Disorder of Piglet Livers: A Proteomic Approach

et al. 2022

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Endoplasmic reticulum stress (ERS) is closely associated with the occurrence and development of many liver diseases. ERS models mostly include experimental animals such as rats and mice. However, pigs are more similar to humans with regards to digestion and metabolism, especially liver construction, yet few reports on ERS in pigs exist. In order to explore changes in the liver under ERS, we used tunicamycin (TM), which can cause liver jaundice and damage liver function, to establish acute ERS models in piglets using a low TM dosage (LD, 0.1 mg/kg body weight (bw)), high TM dosage (HD, 0.3 mg/kg bw), or vehicle for 48 h. We found that both LD- and HD-induced ERS, as verified by the ERS-linked proteins. Furthermore, the concentrations of the proinflammatory cytokines, namely, TNF-α and IL-6 were elevated in TM-treated piglet livers, and the plasma levels of IL-6 and CRP were also higher, indicating the occurrence of inflammation in TM-treated piglets. The complement system was activated in TM-treated piglets, as indicated by increased levels of complement factors and activation products C3, C5a, and AP50. In order to gain insights into the global changes in liver proteins under ERS, we performed an isobaric tag for relative and absolute quantitation (iTRAQ)-based proteomic analysis on the livers of HD- and vehicle-treated piglets. Proteomic analysis identified 311 differentially expressed proteins (DEPs) between the two groups, and a Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis suggested that the DEPs were mainly enriched in signaling pathways such as metabolic pathways, protein processing in the endoplasmic reticulum, and complement and coagulation cascades. Many proteins involved in protein folding, lipid transport, and oxidation were upregulated. Proteins involved in lipid synthesis were downregulated to alleviate liver steatosis, and most complement factors were upregulated to protect the body, and Pearson correlation analysis found that most of the DEPs in the complement and coagulation pathway were significantly correlated with plasma CRP, IL6 and AP50. Our results revealed that TM can activate ERS, marked by liver injury and steatosis, inflammatory reactions, and complement activation in piglets.

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Section: Discussionmentioning

confidence: 99%

Acute Endoplasmic Reticulum Stress Induces Inflammation Reaction, Complement System Activation, and Lipid Metabolism Disorder of Piglet Livers: A Proteomic Approach

et al. 2022

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“…2a, Raman mean spectra of erythrocytes from healthy donors, COVID-19 patients, and convalescent patients show characteristic Raman bands at 1658 (amide I, protein), 1638 (ν(CαCm)asym), 1570 (CβCβ stretching), 1473 (δCH2/ CH3), 1354 (symmetric pyrrole half-ring vibration), 1234 (Methine Cm-H deformation), 1180 (asym. Pyrrole halfring vibration), 1138 (δ(=CbH2)4), 1002 (Phenylalanine), 750 (pyrrole ring def, Tryptophan), 665 (C-S stretch), 551 (S-S stretching), and 396 cm −1 (Fe-N stretching vibrations) [27][28][29][30]. In COVID-19 patients we found a decrease of the 1638 cm −1 and an increase of the 1658 cm −1 bands, indicating a change in the spin state and secondary structure of the hemoglobin.…”

Section: Raman Spectra Of Erythrocytes From Covid-19 Patients Show Si...mentioning

confidence: 99%

SARS-CoV-2 Infects Red Blood Cell Progenitors and Dysregulates Hemoglobin and Iron Metabolism

Kronstein‐Wiedemann

Stadtmüller

Traikov

et al. 2022

Stem Cell Rev and Rep

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Background SARS-CoV-2 infection causes acute respiratory distress, which may progress to multiorgan failure and death. Severe COVID-19 disease is accompanied by reduced erythrocyte turnover, low hemoglobin levels along with increased total bilirubin and ferritin serum concentrations. Moreover, expansion of erythroid progenitors in peripheral blood together with hypoxia, anemia, and coagulopathies highly correlates with severity and mortality. We demonstrate that SARS-CoV-2 directly infects erythroid precursor cells, impairs hemoglobin homeostasis and aggravates COVID-19 disease. Methods Erythroid precursor cells derived from peripheral CD34+ blood stem cells of healthy donors were infected in vitro with SARS-CoV-2 alpha variant and differentiated into red blood cells (RBCs). Hemoglobin and iron metabolism in hospitalized COVID-19 patients and controls were analyzed in plasma-depleted whole blood samples. Raman trapping spectroscopy rapidly identified diseased cells. Results RBC precursors express ACE2 receptor and CD147 at day 5 of differentiation, which makes them susceptible to SARS-CoV-2 infection. qPCR analysis of differentiated RBCs revealed increased HAMP mRNA expression levels, encoding for hepcidin, which inhibits iron uptake. COVID-19 patients showed impaired hemoglobin biosynthesis, enhanced formation of zinc-protoporphyrine IX, heme-CO2, and CO-hemoglobin as well as degradation of Fe-heme. Moreover, significant iron dysmetablolism with high serum ferritin and low serum iron and transferrin levels occurred, explaining disturbances of oxygen-binding capacity in severely ill COVID-19 patients. Conclusions Our data identify RBC precursors as a direct target of SARS-CoV-2 and suggest that SARS-CoV-2 induced dysregulation in hemoglobin- and iron-metabolism contributes to the severe systemic course of COVID-19. This opens the door for new diagnostic and therapeutic strategies. Graphical Abstract

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“…The method was standardized in RBCs of hyperbilirubinemic samples wherein the Raman markers that characterized significant variation from the normal were identified and classified using numerous multivariate analytical tests. A detailed description of the technique is available in an earlier article ( 27 ).…”

Section: Introductionmentioning

confidence: 99%

Micro-Raman spectroscopy study of optically trapped erythrocytes in malaria, dengue and leptospirosis infections

Jacob

Lukose²,

Bankapur³

et al. 2022

Front. Med.

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Malaria, dengue and leptospirosis are three tropical infectious diseases that present with severe hematological derangement causing significant morbidity and mortality, especially during the seasonal monsoons. During the course of these infectious diseases, circulating red blood cells are imperiled to the direct ill-effects of the infectious pathogen in the body as well as to the pro-inflammatory cytokines generated as a consequence of the infection. RBCs when exposed to such inflammatory and/or pathogenic milieu are susceptible to injuries such as RBC programmed eryptosis or RBC programmed necrosis. This research aimed to explore the Raman spectra of live red cells that were extracted from patients infected with malaria, dengue, and leptospirosis. Red cells were optically trapped and micro-Raman probed using a 785 nm Diode laser. RBCs from samples of all three diseases displayed Raman signatures that were significantly altered from the normal/healthy. Distinct spectral markers that were common across all the four groups were obtained from various standardized multivariate analytical methods. Following comprehensive examination of multiple studies, we propose these spectral wavenumbers as “Raman markers of RBC injury.” Findings in our study display that anemia-triggering infections can inflict variations in the healthy status of red cells, easily identifiable by selectively analyzing specific Raman markers. Additionally, this study also highlights relevant statistical tools that can be utilized to study Raman spectral data from biological samples which could help identify the very significant Raman peaks from the spectral band. This approach of RBC analysis can foster a better understanding of red cell behavior and their alterations exhibited in health and disease.

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Micro-Raman Spectroscopy Analysis of Optically Trapped Erythrocytes in Jaundice

Cited by 5 publications

References 41 publications

Acute Endoplasmic Reticulum Stress Induces Inflammation Reaction, Complement System Activation, and Lipid Metabolism Disorder of Piglet Livers: A Proteomic Approach

Acute Endoplasmic Reticulum Stress Induces Inflammation Reaction, Complement System Activation, and Lipid Metabolism Disorder of Piglet Livers: A Proteomic Approach

SARS-CoV-2 Infects Red Blood Cell Progenitors and Dysregulates Hemoglobin and Iron Metabolism

Micro-Raman spectroscopy study of optically trapped erythrocytes in malaria, dengue and leptospirosis infections

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