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Article: Polyethylene glycol-based isolation of urinary extracellular vesicles, an easily adoptable protocol.

Singh, Anula Divyash / Patnam, Sreekanth / Manocha, Anisha / Bashyam, Leena / Rengan, Aravind Kumar / Sasidhar, Manda Venkata

MethodsX

2023 Volume 11, Page(s) 102310

Abstract: Urine is a highly advantageous biological specimen for biomarker research and is a non-invasive source. Most of the urinary biomarkers are non-specific, volatile and need extensive validation before clinical adoption. Extracellular vesicles are secreted ... ...

Abstract	Urine is a highly advantageous biological specimen for biomarker research and is a non-invasive source. Most of the urinary biomarkers are non-specific, volatile and need extensive validation before clinical adoption. Extracellular vesicles are secreted by almost all cells and are involved in homoeostasis, intercellular communication, and cellular processes in healthy and pathophysiological states. Urinary extracellular vesicles (UEVs) are released from the urogenital system and mirror the molecular processes of physiological and pathological states of their source cells. Therefore, UEVs serve as a valuable source of biomarkers for the non-invasive diagnosis of various pathologies. They hold a promising source of multiplex biomarkers suitable for prognosis, diagnosis, and therapy monitoring. UEVs are easily accessible, non-invasive, and suited for longitudinal sampling. Although various techniques are available for isolating UEVs, there is yet to be a consensus on a standard and ideal protocol. We have optimized an efficient, reliable, and easily adoptable polyethylene glycol (PEG) based UEV isolation technique following MISEV guidelines. The method is suitable for various downstream applications of UEVs. This could be a cost-effective, consistent, and accessible procedure for many clinical labs and is most suited for longitudinal analysis. Adopting the protocol will pave the way for establishing UEVs as the ideal biomarker source. •Urine can be collected non-invasively and repeatedly, hence a very useful specimen for biomarker discovery. Urinary EVs (UEVs), derived from urine, offer a stable diagnostic tool, but standardised isolation and analysis approaches are warranted.•To have enough UEVs for any study, large volumes of urine sample are necessary, which limits different isolation methods by cost, yield, and time.•The protocol developed could help researchers by offering a cost-effective and dependable UEV isolation method and may lay the foundation for UEVs adoption in clinical space.
Language	English
Publishing date	2023-08-06
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	2830212-6
ISSN	2215-0161
ISSN	2215-0161
DOI	10.1016/j.mex.2023.102310
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Article ; Online: Polyethylene glycol-based isolation of urinary extracellular vesicles, an easily adoptable protocol

Singh, Anula Divyash / Patnam, Sreekanth / Manocha, Anisha / Bashyam, Leena / Rengan, Aravind Kumar / Sasidhar, Manda Venkata

MethodsX. 2023 Dec., v. 11 p.102310-

2023

Abstract	Urine is a highly advantageous biological specimen for biomarker research and is a non-invasive source. Most of the urinary biomarkers are non-specific, volatile and need extensive validation before clinical adoption. Extracellular vesicles are secreted by almost all cells and are involved in homoeostasis, intercellular communication, and cellular processes in healthy and pathophysiological states. Urinary extracellular vesicles (UEVs) are released from the urogenital system and mirror the molecular processes of physiological and pathological states of their source cells. Therefore, UEVs serve as a valuable source of biomarkers for the non-invasive diagnosis of various pathologies. They hold a promising source of multiplex biomarkers suitable for prognosis, diagnosis, and therapy monitoring. UEVs are easily accessible, non-invasive, and suited for longitudinal sampling. Although various techniques are available for isolating UEVs, there is yet to be a consensus on a standard and ideal protocol. We have optimized an efficient, reliable, and easily adoptable polyethylene glycol (PEG) based UEV isolation technique following MISEV guidelines. The method is suitable for various downstream applications of UEVs. This could be a cost-effective, consistent, and accessible procedure for many clinical labs and is most suited for longitudinal analysis. Adopting the protocol will pave the way for establishing UEVs as the ideal biomarker source. •Urine can be collected non-invasively and repeatedly, hence a very useful specimen for biomarker discovery. Urinary EVs (UEVs), derived from urine, offer a stable diagnostic tool, but standardised isolation and analysis approaches are warranted.•To have enough UEVs for any study, large volumes of urine sample are necessary, which limits different isolation methods by cost, yield, and time.•The protocol developed could help researchers by offering a cost-effective and dependable UEV isolation method and may lay the foundation for UEVs adoption in clinical space.
Keywords	biomarkers ; cell communication ; cost effectiveness ; diagnostic techniques ; isolation techniques ; longitudinal studies ; polyethylene ; polyethylene glycol ; prognosis ; therapeutics ; urine ; Isolation of urinary extracellular vesicles ; Liquid biopsy ; Non-invasive diagnostics ; Renal pathologies ; Urinary extracellular vesicles
Language	English
Dates of publication	2023-12
Publishing place	Elsevier B.V.
Document type	Article ; Online
ZDB-ID	2830212-6
ISSN	2215-0161
ISSN	2215-0161
DOI	10.1016/j.mex.2023.102310
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Article ; Online: Prevalence and Impact of Preexisting Comorbidities on Overall Clinical Outcomes of Hospitalized COVID-19 Patients.

Koyyada, Rajeswari / Nagalla, Balakrishna / Tummala, Anusha / Singh, Anula D / Patnam, Sreekanth / Barigala, Ravikiran / Kandala, Mahati / Krishna, Vamsi / Manda, Sasidhar V

BioMed research international

2022 Volume 2022, Page(s) 2349890

Abstract: COVID-19 risk increases with comorbidities, and the effect is magnified due to the contribution of individual and combined comorbidities to the overall clinical outcomes. We aimed to explore the influence of demographic factors, clinical manifestations, ... ...

Abstract	COVID-19 risk increases with comorbidities, and the effect is magnified due to the contribution of individual and combined comorbidities to the overall clinical outcomes. We aimed to explore the influence of demographic factors, clinical manifestations, and underlying comorbidities on mortality, severity, and hospital stay in COVID-19 patients. Therefore, retrospective chart reviews were performed to identify all laboratory-confirmed cases of SARS-CoV-2 infection in Apollo Hospitals, Hyderabad, between March 2020 and August 2020.A total of 369 confirmed SARS-CoV-2 cases were identified: 272 (73.7%) patients were male, and 97 (26.2%) were female. Of the confirmed cases, 218 (59.1%) had comorbidities, and 151 (40.9%) were devoid of comorbidities. This study showed that old age and underlying comorbidities significantly increase mortality, hospital stay, and severity due to COVID-19 infection. The presence of all four comorbidities, diabetes mellitus (DM) + Hypertension (HTN) + coronary artery disease (CAD) + chronic kidney disease (CKD), conferred the most severity (81%). The highest mortality (OR: 44.03, 95% CI: 8.64-224.27) was observed during the hospital stay (12.73 ± 11.38; 95% CI: 5.08-20.38) in the above group. Multivariate analysis revealed that nonsurvivors are highest (81%) in (DM + HTN + CAD + CKD) category with an odds ratio (95% CI) of 44.03 (8.64-224.27). Age, gender, and comorbidities adjusted odds ratio decreased to 20.25 (3.77-108.77). Median survival of 7 days was observed in the (DM + HTN + CAD + CKD) category. In summary, the presence of underlying comorbidities has contributed to a higher mortality rate, greater risk of severe disease, and extended hospitalization periods, hence, resulting in overall poorer clinical outcomes in hospitalized COVID-19 patients.
MeSH term(s)	COVID-19/epidemiology ; Comorbidity ; Diabetes Mellitus/epidemiology ; Female ; Hospitalization ; Humans ; Hypertension/complications ; Hypertension/epidemiology ; Male ; Prevalence ; Renal Insufficiency, Chronic/epidemiology ; Retrospective Studies ; Risk Factors ; SARS-CoV-2
Language	English
Publishing date	2022-04-06
Publishing country	United States
Document type	Journal Article
ZDB-ID	2698540-8
ISSN	2314-6141 ; 2314-6133
ISSN (online)	2314-6141
ISSN	2314-6133
DOI	10.1155/2022/2349890
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Article ; Online: Differential Expression of SRY-Related HMG-Box Transcription Factor 2, Oligodendrocyte Lineage Transcription Factor 2, and Zinc Finger E-Box Binding Homeobox 1 in Serum-Derived Extracellular Vesicles: Implications for Mithramycin Sensitivity and Targeted Therapy in High-Grade Glioma.

Patnam, Sreekanth / Majumder, Biswanath / Joshi, Parth / Singh, Anula Divyash / Nagalla, Balakrishna / Kumar, Dilli / Biswas, Manjusha / Ranjan, Alok / Majumder, Pradip K / Rengan, Aravind Kumar / Kamath, Ajith V / Ray, Amitava / Manda, Sasidhar Venkata

ACS pharmacology & translational science

2023 Volume 7, Issue 1, Page(s) 137–149

Abstract: Glioblastoma multiforme (GBM) is the most aggressive type of glioma and is often resistant to traditional therapies. Evidence suggests that glioma stem cells (GSCs) contribute to this resistance. Mithramycin (Mit-A) targets GSCs and exhibits antitumor ... ...

Abstract	Glioblastoma multiforme (GBM) is the most aggressive type of glioma and is often resistant to traditional therapies. Evidence suggests that glioma stem cells (GSCs) contribute to this resistance. Mithramycin (Mit-A) targets GSCs and exhibits antitumor activity in GBM by affecting transcriptional targets such as SRY-related HMG-box transcription factor 2 (SOX2), oligodendrocyte lineage transcription factor 2 (OLIG2), and zinc finger E-box binding homeobox 1 (ZEB1). However, its clinical use has been limited by toxicity. This study explored the diagnostic potential of serum extracellular vesicles (EVs) to identify Mit-A responders. Serum EVs were isolated from 70 glioma patients, and targeted gene expression was analyzed using qRT-PCR. Using chemosensitivity assay, we identified 8 Mit-A responders and 17 nonresponders among 25 glioma patients. The M-score showed a significant correlation (
Language	English
Publishing date	2023-12-18
Publishing country	United States
Document type	Journal Article
ISSN	2575-9108
ISSN (online)	2575-9108
DOI	10.1021/acsptsci.3c00198
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Article ; Online: Exploring urinary extracellular vesicles for organ transplant monitoring: A comprehensive study for detection of allograft dysfunction using immune-specific markers.

Singh, Anula Divyash / Nagalla, Balakrishna / Patnam, Sreekanth / Satyanaryana, G / Andrews, Ravi / Panigrahi, Aswini Kumar / Mudigonda, Soma Sekhar / Maitra, Sanjay / Rengan, Aravind Kumar / Sasidhar, Manda Venkata

Clinica chimica acta; international journal of clinical chemistry

2023 Volume 548, Page(s) 117525

Abstract: Background: Allograft dysfunction (AGD) is a common complication following solid organ transplantation (SOT). This study leverages the potential of urinary extracellular vesicles (UEVs) for the non-invasive detection of AGD.: Aim: We aimed to assess ... ...

Abstract	Background: Allograft dysfunction (AGD) is a common complication following solid organ transplantation (SOT). This study leverages the potential of urinary extracellular vesicles (UEVs) for the non-invasive detection of AGD. Aim: We aimed to assess the diagnostic value of T-cell and B-cell markers characteristic of T-cell-mediated and antibody-mediated rejection in UEV-mRNA using renal transplantation as a model. Materials and methods: UEVs were isolated from 123 participants, spanning healthy controls, functional transplant recipients, and biopsy-proven AGD patients. T-cell and B-cell marker mRNA expressions were evaluated using RT-qPCR. Results: We observed significant differences in marker expression between healthy controls and AGD patients. ROC analysis revealed an AUC of 0.80 for T-cell markers, 0.98 for B-cell markers, and 0.94 for combined markers. T-cell markers achieved 81.3 % sensitivity, 80 % specificity, and 80.4 % efficiency. A triad of T-cell markers (PRF1, OX40, and CD3e) increased sensitivity to 87.5 % and efficiency to 82.1 %. B-cell markers (CD20, CXCL3, CD46, and CF3) delivered 100 % sensitivity and 97.5 % specificity. The combined gene signature of T-cell and B-cell markers offered 93.8 % sensitivity and 95 % specificity. Conclusion: Our findings underscore the diagnostic potential of UEV-derived mRNA markers for T-cells and B-cells in AGD, suggesting a promising non-invasive strategy for monitoring graft health.
MeSH term(s)	Humans ; Organ Transplantation ; Transplantation, Homologous ; Extracellular Vesicles ; CD3 Complex ; RNA, Messenger/genetics ; Allografts
Chemical Substances	CD3 Complex ; RNA, Messenger
Language	English
Publishing date	2023-08-24
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	80228-1
ISSN	1873-3492 ; 0009-8981
ISSN (online)	1873-3492
ISSN	0009-8981
DOI	10.1016/j.cca.2023.117525
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Article ; Online: Diagnostic Values of Laboratory Biomarkers in Predicting a Severe Course of COVID-19 on Hospital Admission.

Tummala, Anusha / Ramesh, Venkat / Balakrishna, Nagalla / Koyyada, Rajeswari / Singh, Anula Divyash / Patnam, Sreekanth / Satish Kumar, M / Varahala, Sneha / Manda, Sasidhar V / Narreddy, Suneetha

BioMed research international

2022 Volume 2022, Page(s) 5644956

Abstract: Objective: We intend to identify differences in the clinicodemographic and laboratory findings of COVID-19 patients to predict disease severity and outcome on admission.: Methods: This single-centred retrospective study retrieved laboratory and ... ...

Abstract	Objective: We intend to identify differences in the clinicodemographic and laboratory findings of COVID-19 patients to predict disease severity and outcome on admission. Methods: This single-centred retrospective study retrieved laboratory and clinical data from 350 COVID-19 patients on admission, represented as frequency tables. A multivariate regression model was used to assess the statistically significant association between the explanatory variables and COVID-19 infection outcomes, where adjusted odds ratio (AOR), Results: Among the 350 COVID-19 patients studied, there was a significant increase in the WBC count, neutrophils, aggregate index of systemic inflammation (AISI), neutrophil-to-lymphocyte ratio (dNLR), neutrophil-to-lymphocyte and platelet ratio (NLPR), monocyte-to-lymphocyte ratio (MLR), systemic immune-inflammation index (SII), systemic inflammation response index (SIRI), D-dimer, interleukin-6 (IL-6), ferritin, lactate dehydrogenase (LDH), prothrombin time (PT), glucose, urea, urea nitrogen, creatinine, alanine phosphatase (ALP), and aspartate aminotransferase (AST) and a significant decrease in lymphocytes, eosinophils, total protein, albumin, prealbumin serum, and albumin/globulin (A/G) ratio in the severe group when compared with the mild and moderate groups. However, after adjusting their age, gender, and comorbidities, WBC count (adjusted odds ratio (AOR) = 6.888, 95% CI = 1.590-29.839,
MeSH term(s)	Humans ; COVID-19/diagnosis ; Retrospective Studies ; Biomarkers ; Inflammation ; Neutrophils ; Albumins ; Urea ; Hospitals ; COVID-19 Testing
Chemical Substances	Biomarkers ; Albumins ; Urea (8W8T17847W)
Language	English
Publishing date	2022-11-07
Publishing country	United States
Document type	Journal Article
ZDB-ID	2698540-8
ISSN	2314-6141 ; 2314-6133
ISSN (online)	2314-6141
ISSN	2314-6133
DOI	10.1155/2022/5644956
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Article ; Online: Identifying stable reference genes in polyethene glycol precipitated urinary extracellular vesicles for RT-qPCR-based gene expression studies in renal graft dysfunction patients.

Singh, Anula Divyash / Patnam, Sreekanth / Koyyada, Rajeswari / Samal, Rasmita / Alvi, Syed Baseeruddin / Satyanaryana, G / Andrews, Ravi / Panigrahi, Aswini Kumar / Rengan, Aravind Kumar / Mudigonda, Soma Sekhar / Maitra, Sanjay / Sasidhar, Manda Venkata

Transplant immunology

2022 Volume 75, Page(s) 101715

Abstract: Background: Urinary extracellular vesicles (UEVs) hold RNA in their cargo and are potential sources of biomarkers for gene expression studies. The most used technique for gene-expression studies is quantitative polymerase chain reaction (qPCR). It is ... ...

Abstract	Background: Urinary extracellular vesicles (UEVs) hold RNA in their cargo and are potential sources of biomarkers for gene expression studies. The most used technique for gene-expression studies is quantitative polymerase chain reaction (qPCR). It is critical to use stable reference genes (RGs) as internal controls for normalising gene expression data, which aren't currently available for UEVs. Methods: UEVs were precipitated from urine of graft dysfunction patients and healthy controls by Polyethylene glycol, Mn6000 (PEG6K). Vesicular characterisation confirmed the presence of UEVs. Gene expression levels of five commonly used RGs, i.e., Beta-2-Microglobulin (B2M), ribosomal-protein-L13a (RPL13A), Peptidylprolyl-Isomerase-A (PPIA), hydroxymethylbilane synthase (HMBS), and glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) were quantified, and their stability was established through the RefFinder. The stability of identified RGs was validated by quantification of Perforin and granzyme B, signature molecules of renal graft dysfunction. Results: Urine precipitated with 12% 6 K PEG yielded round and double-membraned UEVs of size ranging from 30 to 100 nm, as confirmed through transmission electron microscopy. Nanoparticle tracking analysis (59 ± 22 nm) and Dynamic-light-scattering (78 ± 56.5 nm) confirmed their size profile. Semi-quantitative Exocheck antibody array demonstrated the presence of EV protein markers in UEV. Using the comparative ΔCт method and RefFinder analysis, B2M (1.6) and RPL13A (1.8) genes emerged as the most stable reference genes. Validation of target gene expression in renal graft dysfunction patients confirmed the efficiency of B2M and RPL13A through significant upregulation compared to other RGs. Conclusions: Our study identified and validated B2M and RPL13A as optimal RGs for mRNA quantification studies in the UEVs of patients with renal graft dysfunction.
MeSH term(s)	Humans ; RNA, Messenger ; Biomarkers/metabolism ; Gene Expression ; Extracellular Vesicles/metabolism ; Polyethylene Glycols ; Real-Time Polymerase Chain Reaction/methods
Chemical Substances	RNA, Messenger ; Biomarkers ; Polyethylene Glycols (3WJQ0SDW1A)
Language	English
Publishing date	2022-09-17
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1160846-8
ISSN	1878-5492 ; 0966-3274
ISSN (online)	1878-5492
ISSN	0966-3274
DOI	10.1016/j.trim.2022.101715
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Article ; Online: Exosomal PTEN as a Predictive Marker of Aggressive Gliomas.

Patnam, Sreekanth / Samal, Rasmita / Koyyada, Rajeswari / Joshi, Partha / Singh, Anula D / Nagalla, Balakrishna / Soma, Madan R / Sannareddy, Rajesh R / Ippili, Kaushal / Raju, Subodh / Boola, Ratnam G / Lath, Rahul / Ranjan, Alok / Ghosh, Siddharth / Balamurugan, M / Ray, Amitava / Manda, Sasidhar V

Neurology India

2022 Volume 70, Issue 1, Page(s) 215–222

Abstract: Background: Liquid biopsies have emerged as convenient alternative diagnostic methods to invasive biopsies, by evaluating disease-specific biomarkers and monitoring the disease risk noninvasively. Phosphatase and tensin homolog deleted in chromosome 10 ( ...

Abstract	Background: Liquid biopsies have emerged as convenient alternative diagnostic methods to invasive biopsies, by evaluating disease-specific biomarkers and monitoring the disease risk noninvasively. Phosphatase and tensin homolog deleted in chromosome 10 (PTEN) is a potent tumor suppressor, and its deletion/mutations are common in gliomas. Objective: Evaluate the feasibility of non-invasive detection of PTEN and its downstream genes in serum exosomes of glioma patients. Materials and methods: PTEN, Yes-associated-protein 1 (YAP1), and lysyl oxidase (LOX) transcript expression were monitored through polymerase chain reaction (PCR) in serum exosomes and their paired tumor tissues. The impact of PTEN and its axis genes expression on the overall survival (OS) was monitored. Results: Out of the 106 glioma serum samples evaluated, PTEN was retained/lost in 65.4%/34.6% of the tumor samples while it was retained/lost in 67.1%/32.9% of their paired exosomal fractions. PTEN expression in both tissue and paired exosomal fractions was observed in 48.11% of the samples. Sanger sequencing detected three mutations (Chr10: 89720791(A>G), Chr10:89720749(C>T), and Chr10:89720850(A>G). Both PTEN-responsive downstream genes (YAP1) and LOX axis were upregulated in the PTEN-deficient samples. PTEN loss was associated with poor survival in the glioma patients (hazard ratio (HR) 0.68, confidence interval (CI): 0.35-1.31, P = 0.28). The OS of the exosomal PTEN cohort coincided with the tumor-tissue PTEN devoid group (HR 1.08, CI: 0.49-2.36, P = 0.85). While, old age yielded the worst prognosis; gender, location, and grade were not prognostic of OS in the multivariate analysis. Conclusions: PTEN and its responsive genes YAP1 and LOX can be detected in serum exosomes and can serve as essential tools for the non-invasive evaluation/identification of aggressive gliomas.
MeSH term(s)	Biomarkers, Tumor ; Brain Neoplasms/diagnosis ; Brain Neoplasms/enzymology ; Brain Neoplasms/genetics ; Brain Neoplasms/pathology ; Glioma/diagnosis ; Glioma/enzymology ; Glioma/genetics ; Glioma/pathology ; Humans ; Mutation ; PTEN Phosphohydrolase/genetics ; Prognosis
Chemical Substances	Biomarkers, Tumor ; PTEN Phosphohydrolase (EC 3.1.3.67) ; PTEN protein, human (EC 3.1.3.67)
Language	English
Publishing date	2022-03-19
Publishing country	India
Document type	Journal Article
ZDB-ID	415522-1
ISSN	1998-4022 ; 0028-3886
ISSN (online)	1998-4022
ISSN	0028-3886
DOI	10.4103/0028-3886.338731
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Article ; Online: Myostatin induces insulin resistance via Casitas B-lineage lymphoma b (Cblb)-mediated degradation of insulin receptor substrate 1 (IRS1) in response to high calorie diet intake.

Bonala, Sabeera / Lokireddy, Sudarsanareddy / McFarlane, Craig / Patnam, Sreekanth / Sharma, Mridula / Kambadur, Ravi

The Journal of biological chemistry

2016 Volume 291, Issue 27, Page(s) 14392

Language	English
Publishing date	2016-05-27
Publishing country	United States
Document type	Journal Article ; Retraction of Publication
ZDB-ID	2997-x
ISSN	1083-351X ; 0021-9258
ISSN (online)	1083-351X
ISSN	0021-9258
DOI	10.1074/jbc.A113.529925
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Article ; Online: Myostatin induces insulin resistance via Casitas B-lineage lymphoma b (Cblb)-mediated degradation of insulin receptor substrate 1 (IRS1) protein in response to high calorie diet intake.

Bonala, Sabeera / Lokireddy, Sudarsanareddy / McFarlane, Craig / Patnam, Sreekanth / Sharma, Mridula / Kambadur, Ravi

publication RETRACTED

The Journal of biological chemistry

2014 Volume 289, Issue 11, Page(s) 7654–7670

Abstract: To date a plethora of evidence has clearly demonstrated that continued high calorie intake leads to insulin resistance and type-2 diabetes with or without obesity. However, the necessary signals that initiate insulin resistance during high calorie intake ...

Abstract	To date a plethora of evidence has clearly demonstrated that continued high calorie intake leads to insulin resistance and type-2 diabetes with or without obesity. However, the necessary signals that initiate insulin resistance during high calorie intake remain largely unknown. Our results here show that in response to a regimen of high fat or high glucose diets, Mstn levels were induced in muscle and liver of mice. High glucose- or fat-mediated induction of Mstn was controlled at the level of transcription, as highly conserved carbohydrate response and sterol-responsive (E-box) elements were present in the Mstn promoter and were revealed to be critical for ChREBP (carbohydrate-responsive element-binding protein) or SREBP1c (sterol regulatory element-binding protein 1c) regulation of Mstn expression. Further molecular analysis suggested that the increased Mstn levels (due to high glucose or fatty acid loading) resulted in increased expression of Cblb in a Smad3-dependent manner. Casitas B-lineage lymphoma b (Cblb) is an ubiquitin E3 ligase that has been shown to specifically degrade insulin receptor substrate 1 (IRS1) protein. Consistent with this, our results revealed that elevated Mstn levels specifically up-regulated Cblb, resulting in enhanced ubiquitin proteasome-mediated degradation of IRS1. In addition, over expression or knock down of Cblb had a major impact on IRS1 and pAkt levels in the presence or absence of insulin. Collectively, these observations strongly suggest that increased glucose levels and high fat diet, both, result in increased circulatory Mstn levels. The increased Mstn in turn is a potent inducer of insulin resistance by degrading IRS1 protein via the E3 ligase, Cblb, in a Smad3-dependent manner.
MeSH term(s)	Animals ; Base Sequence ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ; Blood Glucose/metabolism ; Diabetes Mellitus, Type 2/metabolism ; Diet/adverse effects ; Dietary Fats/administration & dosage ; Gene Expression Regulation ; Glucose/administration & dosage ; Glucose/metabolism ; Hep G2 Cells ; Humans ; Insulin Receptor Substrate Proteins/metabolism ; Insulin Resistance ; Lentivirus/metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Molecular Sequence Data ; Muscle, Skeletal/metabolism ; Myostatin/metabolism ; Nuclear Proteins/metabolism ; Palmitates/metabolism ; Proto-Oncogene Proteins c-cbl/metabolism ; Smad3 Protein/metabolism ; Sterol Regulatory Element Binding Protein 1/metabolism ; Transcription Factors/metabolism
Chemical Substances	Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ; Blood Glucose ; Dietary Fats ; Insulin Receptor Substrate Proteins ; Irs1 protein, mouse ; Mlxipl protein, mouse ; Myostatin ; Nuclear Proteins ; Palmitates ; Smad3 Protein ; Smad3 protein, mouse ; Srebf1 protein, mouse ; Sterol Regulatory Element Binding Protein 1 ; Transcription Factors ; Proto-Oncogene Proteins c-cbl (EC 2.3.2.27) ; Cbl protein, mouse (EC 6.3.2.-) ; Glucose (IY9XDZ35W2)
Language	English
Publishing date	2014-01-22
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Retracted Publication
ZDB-ID	2997-x
ISSN	1083-351X ; 0021-9258
ISSN (online)	1083-351X
ISSN	0021-9258
DOI	10.1074/jbc.M113.529925
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