LC-MS Based Platform Simplifies Access to Metabolomics for Peroxisomal Disorders

Peroxisomes are central hubs for cell metabolism and their dysfunction is linked to devastating human disorders, such as peroxisomal biogenesis disorders and single peroxisomal enzyme/protein deficiencies. For decades, biochemical diagnostics have been carried out using classical markers such as very long-chain fatty acids (VLCFA), which can be inconspicuous in milder and atypical cases. Holistic metabolomics studies revealed several potentially new biomarkers for peroxisomal disorders for advanced laboratory diagnostics including atypical cases. However, establishing these new markers is a major challenge in routine diagnostic laboratories. We therefore investigated whether the commercially available AbsoluteIDQ p180 kit (Biocrates Lifesciences), which utilizes flow injection and liquid chromatography mass spectrometry, may be used to reproduce some key results from previous global metabolomics studies. We applied it to serum samples from patients with mutations in peroxisomal target genes PEX1, ABCD1, and the HSD17B4 gene. Here we found various changes in sphingomyelins and lysophosphatidylcholines. In conclusion, this kit can be used to carry out extended diagnostics for peroxisomal disorders in routine laboratories, even without access to a metabolomics unit.

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Peroxisomal biogenesis disorders in Zellweger syndrome spectrum: diagnosis, monitoring and treatment according to the recommendations of the Global Foundation for Peroxisomal Disorders

CHILD`S HEALTH ◽

10.22141/2224-0551.13.2.2018.129554 ◽

2018 ◽

Vol 13 (2) ◽

pp. 194-203

Author(s):

M.A. Gonchar ◽

G.R. Muratov ◽

O.L. Logvinova ◽

E.M. Pushkar ◽

E.P. Pomazunovskaya ◽

...

Keyword(s):

Zellweger Syndrome ◽

Peroxisomal Disorders ◽

Peroxisomal Biogenesis ◽

Peroxisomal Biogenesis Disorders

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Urine acylcarnitine analysis by ESI–MS/MS: A new tool for the diagnosis of peroxisomal biogenesis disorders

Clinica Chimica Acta ◽

10.1016/j.cca.2008.08.018 ◽

2008 ◽

Vol 398 (1-2) ◽

pp. 86-89 ◽

Cited By ~ 14

Author(s):

Guglielmo Duranti ◽

Sara Boenzi ◽

Cristiano Rizzo ◽

Lucilla Ravà ◽

Vincenzo Di Ciommo ◽

...

Keyword(s):

Esi Ms ◽

Peroxisomal Biogenesis ◽

Acylcarnitine Analysis ◽

Peroxisomal Biogenesis Disorders

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Clinical and diagnostic manifestations of tickborne mixed infection in combination with COVID-19

Russian Clinical Laboratory Diagnostics ◽

10.51620/0869-2084-2021-66-11-689-694 ◽

2021 ◽

Vol 66 (11) ◽

pp. 689-694

Author(s):

A. L. Shutikova ◽

G. N. Leonova ◽

A. F. Popov ◽

M. Yu. Shchelkanov

Keyword(s):

Lyme Disease ◽

Erythema Migrans ◽

Clinical Manifestations ◽

Underlying Disease ◽

Laboratory Diagnostics ◽

Igg Antibodies ◽

Serum Samples ◽

Igm Antibodies ◽

Chronic Encephalitis ◽

Tbev Strain

The coexistence of various pathogens inside the patient’s body is one of the poorly studied and current issues. The aim of the study is to identify the relationship between the indicators of complex laboratory diagnostics and the clinical manifestations of a mixed disease during subsequent infection with the SARS-CoV-2 virus using the example of a case of chronic encephalitis-borreliosis infection. Seven blood serum samples were collected from the patient over the course of a year. For the etiological verification of the causative agents of TBE, Lyme disease and COVID-19, the methods of ELISA and PCR diagnostics were used. The patient was diagnosed with Lyme disease on the basis of the detection of IgG antibodies to Borrelia 5 months after the onset of the disease, since she denied the tick bite. In the clinical picture, there was an articular syndrome and erythema migrans. Later, IgG antibodies to the TBEV were found in the blood. Throughout the study, IgM antibodies to Borrelia were not detected. The exacerbation of Lyme disease could be judged by the clinical manifestations of this disease and by the growth of specific IgG antibodies. A feature of this case was that during an exacerbation of the Lyme disease, an infection with the SARS-CoV-2 virus occurred. Treatment (umifenovir, hydroxychloroquine, azithromycin, ceftriaxone) was prescribed, which improved the condition of the underlying disease, decreased joint pain, decreased IgG levels to borrelia. However, during this period, serological markers of TBEV appear: antigen, IgM antibodies, and the titer of IgG antibodies increases. Most likely, this was facilitated by the switching of the immune system to the SARS-CoV-2 virus, with the simultaneous suppression of borrelia with antibiotics and the appointment of hydroxychloroquine, which has an immunosuppressive effect. Despite the activation of the virus, clinical manifestations of TBE were not observed in the patient, which is most likely associated with infection with a weakly virulent TBEV strain. The further course of tick-borne infections revealed the dominant influence of B. burgdorferi in relation to TBEV. Laboratory studies have shown that suppression of the activity of the borreliosis process by etiotropic treatment subsequently led to the activation of the persistent TBEV.

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Clinical Relevance of Serum Galactose Deficient IgA1 in Patients with IgA Nephropathy

Journal of Clinical Medicine ◽

10.3390/jcm9113549 ◽

2020 ◽

Vol 9 (11) ◽

pp. 3549

Author(s):

Jin Sug Kim ◽

Hyeon Seok Hwang ◽

Sang Ho Lee ◽

Yang Gyun Kim ◽

Ju-Young Moon ◽

...

Keyword(s):

Iga Nephropathy ◽

Clinical Relevance ◽

Interstitial Fibrosis ◽

Healthy Controls ◽

Ckd Progression ◽

Serum Samples ◽

Tubular Atrophy ◽

Appropriate Treatment ◽

Cox Proportional Hazard Models ◽

New Biomarkers

New biomarkers of IgA nephropathy (IgAN) are needed for non-invasive diagnosis and appropriate treatment. There is emerging evidence that galactose deficient IgA1 (Gd-IgA1) is a pivotal molecule in the pathogenesis of IgAN. However, few studies have investigated the role of Gd-IgA1 as a biomarker in IgAN. In this study, we investigated the clinical relevance of serum Gd-IgA1 levels in patients with IgAN. Two hundred and thirty biopsy-proven IgAN patients, 74 disease controls (patients with non-IgAN nephropathy), and 15 healthy controls were enrolled in this study. Levels of serum Gd-IgA1 were measured using an ELISA kit in serum samples obtained the day of renal biopsy. We compared levels of serum Gd-IgA1 according to the type of glomerular disease and analyzed the association between Gd-IgA1 levels and clinical and pathological parameters in patients with IgAN. We then divided IgAN patients into two groups according to Gd-IgA1 level and investigated the predictive value of Gd-IgA1 for progression of chronic kidney disease (CKD). Serum Gd-IgA1 levels were significantly higher in IgAN patients than disease controls and healthy controls. In patients with IgAN, serum Gd-IA1 levels were significantly correlated with estimated glomerular filtration rate, serum IgA level, and tubular atrophy/interstitial fibrosis. CKD progression was more frequent in IgAN patients with higher serum Gd-IgA1 levels than in those with lower serum Gd-IgA1 levels. Cox proportional hazard models showed that high GdIgA1 level was an independent risk factor for CKD progression after adjusting for several confounders. Our results suggest that serum Gd-IgA1 level is a useful diagnostic and prognostic marker in IgAN patients. Further studies with a larger sample size and longer follow-up duration are needed.

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Circulating miRNAs as a Predictive Biomarker of the Progression from Prediabetes to Diabetes: Outcomes of a 5-Year Prospective Observational Study

Journal of Clinical Medicine ◽

10.3390/jcm9072184 ◽

2020 ◽

Vol 9 (7) ◽

pp. 2184 ◽

Cited By ~ 1

Author(s):

Iwona Sidorkiewicz ◽

Magdalena Niemira ◽

Katarzyna Maliszewska ◽

Anna Erol ◽

Agnieszka Bielska ◽

...

Keyword(s):

Pathway Analysis ◽

Mirna Target ◽

Target Genes ◽

Quantitative Polymerase Chain Reaction ◽

Area Under The Curve ◽

Predictive Biomarkers ◽

Predictive Biomarker ◽

Circulating Mirnas ◽

Serum Samples ◽

Roc Curve Analysis

Due to a global increase in the prevalence of type 2 diabetes mellitus (T2DM), there is an urgent need for early identification of prediabetes, as these people have the highest risk of developing diabetes. Circulating miRNAs have shown potential as progression biomarkers in other diseases. This study aimed to conduct a baseline comparison of serum-circulating miRNAs in prediabetic individuals, with the distinction between those who later progressed to T2DM and those who did not. The expression levels of 798 miRNAs using NanoString technology were examined. Spearman correlation, receiver operating characteristic (ROC) curve analysis, and logistic regression modeling were performed. Gene ontology (GO) and canonical pathway analysis were used to explore the biological functions of the miRNA target genes. The study revealed that three miRNAs were upregulated in the serum samples of patients who later progressed to T2DM. Pathway analysis showed that the miRNA target genes were mainly significantly enriched in neuronal NO synthase (nNOS) signaling in neurons, amyloid processing, and hepatic cholestasis. ROC analysis demonstrated that miR-491-5p, miR-1307-3p, and miR-298 can be introduced as a diagnostic tool for the prediction of T2DM (area under the curve (AUC) = 94.0%, 88.0%, and 84.0%, respectively). Validation by real-time quantitative polymerase chain reaction (qRT-PCR) confirmed our findings. The results suggest that circulating miRNAs can potentially be used as predictive biomarkers of T2DM in prediabetic patients.

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Peroxisome Metabolism in Cancer

Cells ◽

10.3390/cells9071692 ◽

2020 ◽

Vol 9 (7) ◽

pp. 1692 ◽

Cited By ~ 1

Author(s):

Jung-Ae Kim

Keyword(s):

Metabolic Pathways ◽

Cancer Metabolism ◽

Cell Metabolism ◽

Critical Role ◽

Acid Oxidation ◽

Metabolic Dysregulation ◽

Human Disorders ◽

Peroxisomal Function ◽

Target Cancer

Peroxisomes are metabolic organelles involved in lipid metabolism and cellular redoxbalance. Peroxisomal function is central to fatty acid oxidation, ether phospholipid synthesis, bile acidsynthesis, and reactive oxygen species homeostasis. Human disorders caused by genetic mutations inperoxisome genes have led to extensive studies on peroxisome biology. Peroxisomal defects are linkedto metabolic dysregulation in diverse human diseases, such as neurodegeneration and age-relateddisorders, revealing the significance of peroxisome metabolism in human health. Cancer is a diseasewith metabolic aberrations. Despite the critical role of peroxisomes in cell metabolism, the functionaleects of peroxisomes in cancer are not as well recognized as those of other metabolic organelles,such as mitochondria. In addition, the significance of peroxisomes in cancer is less appreciated thanit is in degenerative diseases. In this review, I summarize the metabolic pathways in peroxisomesand the dysregulation of peroxisome metabolism in cancer. In addition, I discuss the potential ofinactivating peroxisomes to target cancer metabolism, which may pave the way for more eectivecancer treatment.

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Peroxisomal Disorders

10.1093/med/9780199937837.003.0069 ◽

2017 ◽

Author(s):

Gerald V. Raymond ◽

Mohamed Y. Jefri ◽

Kristin W. Baranano ◽

Ali Fatemi

Keyword(s):

Peroxisomal Disorder ◽

Peroxisomal Disorders ◽

Peroxisomal Enzyme ◽

Second Category

Disorders of the peroxisome are divided into two major categories. In the first, the organelle fails to develop normally, leading to disruption of multiple peroxisomal enzymes. The second category consists of those disorders in which the peroxisome structure is normal but functioning of a single peroxisomal enzyme or protein is defective. While there is an expanding list of disorders in both categories, this chapter focuses on X-linked adrenoleukodystrophy, the most common peroxisomal disorder, and on peroxisomal assembly/biogenesis disorders.

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The Peroxisomal PTS1-Import Defect of PEX1- Deficient Cells Is Independent of Pexophagy in Saccharomyces cerevisiae

International Journal of Molecular Sciences ◽

10.3390/ijms21030867 ◽

2020 ◽

Vol 21 (3) ◽

pp. 867 ◽

Cited By ~ 1

Author(s):

Thomas Mastalski ◽

Rebecca Brinkmeier ◽

Harald W. Platta

Keyword(s):

Autosomal Recessive ◽

Matrix Protein ◽

Protein Import ◽

Beta Oxidation ◽

Working Models ◽

Peroxisomal Biogenesis ◽

Physiologic Role ◽

Peroxisomal Biogenesis Disorders ◽

Peroxisomal Function

The important physiologic role of peroxisomes is shown by the occurrence of peroxisomal biogenesis disorders (PBDs) in humans. This spectrum of autosomal recessive metabolic disorders is characterized by defective peroxisome assembly and impaired peroxisomal functions. PBDs are caused by mutations in the peroxisomal biogenesis factors, which are required for the correct compartmentalization of peroxisomal matrix enzymes. Recent work from patient cells that contain the Pex1(G843D) point mutant suggested that the inhibition of the lysosome, and therefore the block of pexophagy, was beneficial for peroxisomal function. The resulting working model proposed that Pex1 may not be essential for matrix protein import at all, but rather for the prevention of pexophagy. Thus, the observed matrix protein import defect would not be caused by a lack of Pex1 activity, but rather by enhanced removal of peroxisomal membranes via pexophagy. In the present study, we can show that the specific block of PEX1 deletion-induced pexophagy does not restore peroxisomal matrix protein import or the peroxisomal function in beta-oxidation in yeast. Therefore, we conclude that Pex1 is directly and essentially involved in peroxisomal matrix protein import, and that the PEX1 deletion-induced pexophagy is not responsible for the defect in peroxisomal function. In order to point out the conserved mechanism, we discuss our findings in the context of the working models of peroxisomal biogenesis and pexophagy in yeasts and mammals.

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Evaluation of reactivity to Echinococcus spp. among rural inhabitants in Poland

Acta Parasitologica ◽

10.1515/ap-2015-0074 ◽

2015 ◽

Vol 60 (3) ◽

Cited By ~ 5

Author(s):

Ewa Cisak ◽

Jacek Sroka ◽

Angelina Wójcik-Fatla ◽

Violetta Zając ◽

Jacek Dutkiewicz

Keyword(s):

Western Blot ◽

Echinococcus Granulosus ◽

Cross Reactivity ◽

Farm Animals ◽

Control Group ◽

Laboratory Diagnostics ◽

Group 14 ◽

Serum Samples ◽

The City ◽

Echinococcus Spp

AbstractA group of 172 rural inhabitants from eastern Poland (68 males and 104 females, mean age 49.0 ± 12.0 years) was examined for the presence of antibodies against Echinococcus granulosus and Echinococcus multilocularis. A population of 38 healthy urban dwellers from the city of Lublin (17 males and 21 females, mean age 36.2 ± 9.6 years) were examined as a control group. Sera of 22 rural inhabitants (12.8%) reacted positively to Echinococcus granulosus hydatid fluid antigen in the screening test. A cross-reactivity was observed with two serum samples that tested positive in ELISA for E. granulosus. Three serum samples were tested positive for E. multilocularis using the Em2plus ELISA assay and also positive for Western blot. None of the members of control group showed the presence of a seropositive reaction to Echinococcus spp. The reactivity to Echinococcus spp. among rural inhabitants decreased with age and this correlation was statistically significant (R = -0.197151, p = 0.009535). The percentage of positive findings was the highest (50.0%) in the youngest age group (14-20). No significant correlations were found between responses to interview questions (possession of domestic and farm animals, contact with wild animals, eating unwashed berries, drinking unboiled water) and the presence of seropositive reactions to Echinococcus spp. The presented results seem to indicate that echinococcosis is still a current problem in Poland that should not be neglected and, moreover, indicates the need for improvement in the routine laboratory diagnostics of Echinococcus spp. by standardizing the ELISA and Western blot tests.

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