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Artikel ; Online: Lyophilization of Molecular Biology Reactions: A Review.

Prado, Nayra Oliveira / Lalli, Larissa Araújo / Blanes, Lucas / Zanette, Dalila Lucíola / Aoki, Mateus Nóbrega

2022 Band 23, Heft 4, Seite(n) 480–496

Abstract: Molecular biology is a widely used and widespread technique in research and as a laboratory diagnostic tool, aiming to investigate targets of interest from the obtainment, identification, and analysis of genetic material. In this context, methods, such ... ...

Abstract	Molecular biology is a widely used and widespread technique in research and as a laboratory diagnostic tool, aiming to investigate targets of interest from the obtainment, identification, and analysis of genetic material. In this context, methods, such as Polymerase Chain Reaction (PCR), Reverse Transcription Polymerase Chain Reaction (RT-PCR), real-time PCR, loopmediated isothermal amplification (LAMP), and loop-mediated isothermal amplification with reverse transcription (RT-LAMP), can be cited. Such methods use enzymes, buffers, and thermosensitive reagents, which require specific storage conditions. In an attempt to solve this problem, the lyophilization procedure (dehydration process by sublimation) can be applied, aiming to preserve and prolong the useful life of the reaction components in cases of temperature variation. In this review, we present a synthesis of the lyophilization process, describing the events of each step of the procedure and providing general information about the technique. Moreover, we selected lyophilization protocols found in the literature, paying attention to the conditions chosen by the authors for each step of the procedure, and structured the main data in tables, facilitating access to information for researchers who need material to produce new functional protocols.
Mesh-Begriff(e)	Humans ; Molecular Biology/instrumentation ; Molecular Biology/methods ; Water/chemistry ; Freeze Drying/instrumentation ; Freeze Drying/methods ; Polymerase Chain Reaction/instrumentation ; Polymerase Chain Reaction/methods ; Cryopreservation ; Point-of-Care Systems
Chemische Substanzen	Water (059QF0KO0R)
Sprache	Englisch
Erscheinungsdatum	2022-08-05
Erscheinungsland	Netherlands
Dokumenttyp	Review ; Journal Article
ZDB-ID	2104081-3
ISSN	1875-5607 ; 1389-5575
ISSN (online)	1875-5607
ISSN	1389-5575
DOI	10.2174/1389557522666220802144057
Datenquelle	MEDical Literature Analysis and Retrieval System OnLINE

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Artikel: Circulating Cell-Free Nucleic Acids as Biomarkers for Diagnosis and Prognosis of Pancreatic Cancer.

Marin, Anelis Maria / Sanchuki, Heloisa Bruna Soligo / Namur, Guilherme Naccache / Uno, Miyuki / Zanette, Dalila Luciola / Aoki, Mateus Nóbrega

Biomedicines

2023 Band 11, Heft 4

Abstract: A lack of reliable early diagnostic tools represents a major challenge in the management of pancreatic cancer (PCa), as the disease is often only identified after it reaches an advanced stage. This highlights the urgent need to identify biomarkers that ... ...

Abstract	A lack of reliable early diagnostic tools represents a major challenge in the management of pancreatic cancer (PCa), as the disease is often only identified after it reaches an advanced stage. This highlights the urgent need to identify biomarkers that can be used for the early detection, staging, treatment monitoring, and prognosis of PCa. A novel approach called liquid biopsy has emerged in recent years, which is a less- or non-invasive procedure since it focuses on plasmatic biomarkers such as DNA and RNA. In the blood of patients with cancer, circulating tumor cells (CTCs) and cell-free nucleic acids (cfNAs) have been identified such as DNA, mRNA, and non-coding RNA (miRNA and lncRNA). The presence of these molecules encouraged researchers to investigate their potential as biomarkers. In this article, we focused on circulating cfNAs as plasmatic biomarkers of PCa and analyzed their advantages compared to traditional biopsy methods.
Sprache	Englisch
Erscheinungsdatum	2023-04-01
Erscheinungsland	Switzerland
Dokumenttyp	Journal Article ; Review
ZDB-ID	2720867-9
ISSN	2227-9059
ISSN	2227-9059
DOI	10.3390/biomedicines11041069
Datenquelle	MEDical Literature Analysis and Retrieval System OnLINE

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Artikel ; Online: microRNA-based Genetic Therapy in Leukemia: Properties, Delivery, and Experimental Models.

Prado, Nayra Oliveira / Kusma Wosniaki, Denise / Maria Marin, Anelis / Mathias, Carolina / Soligo Sanchuki, Heloisa Bruna / Zanette, Dalila Luciola / Nóbrega Aoki, Mateus

Current gene therapy

2023 Band 23, Heft 4, Seite(n) 245–260

Abstract: Leukemia is a type of cancer that affects white blood cells. In this disease, immature blood cells undergo genetic mutations, leading to excessive replication and reduced cell death compared to healthy cells. In cancer, there may be the activation of ... ...

Abstract	Leukemia is a type of cancer that affects white blood cells. In this disease, immature blood cells undergo genetic mutations, leading to excessive replication and reduced cell death compared to healthy cells. In cancer, there may be the activation of oncogenes and the deactivation of tumor suppressor genes that control certain cellular functions. Despite the undeniable contribution to the patient's recovery, conventional cancer treatments may have some not-so-beneficial effects. In this case, gene therapy appears as an alternative to classical treatments. Gene therapy delivers genetic material to cells to replace or modify dysfunctional genes, a safe method for neoplasms. One of the types of nucleic acids explored in gene therapy is microRNA (miRNA), a group of endogenous, non-proteincoding, small single-stranded RNA molecules involved in the regulation of gene expression, cell division, differentiation, angiogenesis, migration, apoptosis, and carcinogenesis. This review aims to bring together the most recent advances found in the literature on cancer gene therapy based on microRNAs in the oncological context, focusing on leukemia.
Mesh-Begriff(e)	Humans ; MicroRNAs/genetics ; MicroRNAs/therapeutic use ; MicroRNAs/metabolism ; Neoplasms/genetics ; Neoplasms/therapy ; Neoplasms/pathology ; Leukemia/genetics ; Leukemia/therapy ; Genetic Therapy/methods ; Models, Theoretical ; Gene Expression Regulation, Neoplastic
Chemische Substanzen	MicroRNAs
Sprache	Englisch
Erscheinungsdatum	2023-06-21
Erscheinungsland	United Arab Emirates
Dokumenttyp	Review ; Journal Article
ZDB-ID	2146187-9
ISSN	1875-5631 ; 1566-5232
ISSN (online)	1875-5631
ISSN	1566-5232
DOI	10.2174/1566523223666230426153622
Datenquelle	MEDical Literature Analysis and Retrieval System OnLINE

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Artikel ; Online: The Screening of microRNAs in Chronic Myeloid Leukemia: A Clinical Evaluation.

Wosniaki, Denise Kusma / Marin, Anelis Maria / Oliveira, Rafaela Noga / Koerich, Gabriela Marino / Munhoz, Eduardo Cilião / Farias, João Samuel de Holanda / Beltrame, Miriam Perlingeiro / Zanette, Dalila Luciola / Aoki, Mateus Nóbrega

International journal of molecular sciences

2024 Band 25, Heft 6

Abstract: Chronic myeloid leukemia (CML) is a type of leukemia whose main genetic marker is the reciprocal translocation that leads to the production of ... ...

Abstract	Chronic myeloid leukemia (CML) is a type of leukemia whose main genetic marker is the reciprocal translocation that leads to the production of the
Mesh-Begriff(e)	Humans ; MicroRNAs/genetics ; Fusion Proteins, bcr-abl/genetics ; Fusion Proteins, bcr-abl/metabolism ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive/diagnosis ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive/genetics ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive/pathology ; Translocation, Genetic ; Biomarkers
Chemische Substanzen	MicroRNAs ; Fusion Proteins, bcr-abl (EC 2.7.10.2) ; Biomarkers ; MIRN7 microRNA, human
Sprache	Englisch
Erscheinungsdatum	2024-03-16
Erscheinungsland	Switzerland
Dokumenttyp	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms25063363
Datenquelle	MEDical Literature Analysis and Retrieval System OnLINE

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Artikel ; Online: Development and evaluation of a lyophilization protocol for colorimetric RT-LAMP diagnostic assay for COVID-19.

Prado, Nayra Oliveira / Marin, Anelis Maria / Lalli, Larissa Araujo / Sanchuki, Heloisa Bruna Soligo / Wosniaki, Denise Kusma / Nardin, Jeanine Marie / Morales, Hugo Manoel Paz / Blanes, Lucas / Zanette, Dalila Luciola / Aoki, Mateus Nóbrega

Scientific reports

2024 Band 14, Heft 1, Seite(n) 10612

Abstract: Molecular diagnostics involving nucleic acids (DNA and RNA) are regarded as extremely functional tools. During the 2020 global health crisis, efforts intensified to optimize the production and delivery of molecular diagnostic kits for detecting SARS-CoV- ... ...

Abstract	Molecular diagnostics involving nucleic acids (DNA and RNA) are regarded as extremely functional tools. During the 2020 global health crisis, efforts intensified to optimize the production and delivery of molecular diagnostic kits for detecting SARS-CoV-2. During this period, RT-LAMP emerged as a significant focus. However, the thermolability of the reagents used in this technique necessitates special low-temperature infrastructure for transport, storage, and conservation. These requirements limit distribution capacity and necessitate cost-increasing adaptations. Consequently, this report details the development of a lyophilization protocol for reagents in a colorimetric RT-LAMP diagnostic kit to detect SARS-CoV-2, facilitating room-temperature transport and storage. We conducted tests to identify the ideal excipients that maintain the molecular integrity of the reagents and ensure their stability during room-temperature storage and transport. The optimal condition identified involved adding 5% PEG 8000 and 75 mM trehalose to the RT-LAMP reaction, which enabled stability at room temperature for up to 28 days and yielded an analytical and diagnostic sensitivity and specificity of 83.33% and 90%, respectively, for detecting SARS-CoV-2. This study presents the results of a lyophilized colorimetric RT-LAMP COVID-19 detection assay with diagnostic sensitivity and specificity comparable to RT-qPCR, particularly in samples with high viral load.
Mesh-Begriff(e)	Freeze Drying ; Humans ; COVID-19/diagnosis ; COVID-19/virology ; SARS-CoV-2/isolation & purification ; SARS-CoV-2/genetics ; Colorimetry/methods ; Nucleic Acid Amplification Techniques/methods ; Molecular Diagnostic Techniques/methods ; RNA, Viral/analysis ; RNA, Viral/genetics ; Sensitivity and Specificity ; Reagent Kits, Diagnostic/standards ; COVID-19 Nucleic Acid Testing/methods
Chemische Substanzen	RNA, Viral ; Reagent Kits, Diagnostic
Sprache	Englisch
Erscheinungsdatum	2024-05-09
Erscheinungsland	England
Dokumenttyp	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-024-61163-7
Datenquelle	MEDical Literature Analysis and Retrieval System OnLINE

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Artikel: Sample Preparation for Lab-on-a-Chip Systems in Molecular Diagnosis: A Review

Cunha, Mylena Lemes / da Silva, Stella Schuster / Stracke, Mateus Cassaboni / Zanette, Dalila Luciola / Aoki, Mateus Nóbrega / Blanes, Lucas

Analytical chemistry. 2021 Dec. 06, v. 94, no. 1

2021

Abstract: Rapid and low-cost molecular analysis is especially required for early and specific diagnostics, quick decision-making, and sparing patients from unnecessary tests and hospitals from extra costs. One way to achieve this objective is through automated ... ...

Abstract	Rapid and low-cost molecular analysis is especially required for early and specific diagnostics, quick decision-making, and sparing patients from unnecessary tests and hospitals from extra costs. One way to achieve this objective is through automated molecular diagnostic devices. Thus, sample-to-answer microfluidic devices are emerging with the promise of delivering a complete molecular diagnosis system that includes nucleic acid extraction, amplification, and detection steps in a single device. The biggest issue in such equipment is the extraction process, which is normally laborious and time-consuming but extremely important for sensitive and specific detection. Therefore, this Review focuses on automated or semiautomated extraction methodologies used in lab-on-a-chip devices. More than 15 different extraction methods developed over the past 10 years have been analyzed in terms of their advantages and disadvantages to improve extraction procedures in future studies. Herein, we are able to explain the high applicability of the extraction methodologies due to the large variety of samples in which different techniques were employed, showing that their applications are not limited to medical diagnosis. Moreover, we are able to conclude that further research in the field would be beneficial because the methodologies presented can be affordable, portable, time efficient, and easily manipulated, all of which are strong qualities for point-of-care technologies.
Schlagwörter	analytical chemistry ; decision making ; diagnostic techniques ; equipment ; lab-on-a-chip technology ; nucleic acids ; point-of-care systems
Sprache	Englisch
Erscheinungsverlauf	2021-1206
Umfang	p. 41-58.
Erscheinungsort	American Chemical Society
Dokumenttyp	Artikel
ZDB-ID	1508-8
ISSN	1520-6882 ; 0003-2700
ISSN (online)	1520-6882
ISSN	0003-2700
DOI	10.1021/acs.analchem.1c04460
Datenquelle	NAL Katalog (AGRICOLA)

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Artikel ; Online: Plasma Exosome-Derived microRNAs as Potential Diagnostic and Prognostic Biomarkers in Brazilian Pancreatic Cancer Patients.

Marin, Anelis Maria / Mattar, Sibelle Botogosque / Amatuzzi, Rafaela Ferreira / Chammas, Roger / Uno, Miyuki / Zanette, Dalila Luciola / Aoki, Mateus Nóbrega

Biomolecules

2022 Band 12, Heft 6

Abstract: Pancreatic cancer represents one of the leading causes of oncological death worldwide. A combination of pancreatic cancer aggressiveness and late diagnosis are key factors leading to a low survival rate and treatment inefficiency, and early diagnosis is ... ...

Abstract	Pancreatic cancer represents one of the leading causes of oncological death worldwide. A combination of pancreatic cancer aggressiveness and late diagnosis are key factors leading to a low survival rate and treatment inefficiency, and early diagnosis is pursued as a critical factor for pancreatic cancer. In this context, plasma microRNAs are emerging as promising players due to their non-invasive and practical usage in oncological diagnosis and prognosis. Recent studies have showed some miRNAs associated with pancreatic cancer subtypes, or with stages of the disease. Here we demonstrate plasma exosome-derived microRNA expression in pancreatic cancer patients and healthy individuals from Brazilian patients. Using plasma of 65 pancreatic cancer patients and 78 healthy controls, plasma exosomes were isolated and miRNAs miR-27b, miR-125b-3p, miR-122-5p, miR-21-5p, miR-221-3p, miR-19b, and miR-205-5p were quantified by RT-qPCR. We found that miR-125b-3p, miR-122-5p, and miR-205-5p were statistically overexpressed in the plasma exosomes of pancreatic cancer patients compared to healthy controls. Moreover, miR-205-5p was significantly overexpressed in European descendants, in patients with tumor progression and in those who died from the disease, and diagnostic ability by ROC curve was 0.86. Therefore, we demonstrate that these three microRNAs are potential plasma exosome-derived non-invasive biomarkers for the diagnosis and prognosis of Brazilian pancreatic cancer, demonstrating the importance of different populations and epidemiological bias.
Mesh-Begriff(e)	Biomarkers/metabolism ; Biomarkers, Tumor/metabolism ; Brazil ; Early Detection of Cancer ; Exosomes/genetics ; Exosomes/metabolism ; Humans ; MicroRNAs/metabolism ; Pancreatic Neoplasms/diagnosis ; Pancreatic Neoplasms/genetics ; Prognosis ; Pancreatic Neoplasms
Chemische Substanzen	Biomarkers ; Biomarkers, Tumor ; MicroRNAs
Sprache	Englisch
Erscheinungsdatum	2022-05-31
Erscheinungsland	Switzerland
Dokumenttyp	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2701262-1
ISSN	2218-273X ; 2218-273X
ISSN (online)	2218-273X
ISSN	2218-273X
DOI	10.3390/biom12060769
Datenquelle	MEDical Literature Analysis and Retrieval System OnLINE

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Artikel ; Online: Germline variants in early and late-onset Brazilian prostate cancer patients.

Coelho, Karoline Brito Caetano Andrade / Squire, Jeremy A / Duarte, Kelly Gomes / Sares, Cláudia Tarcila Gomes / Moreda, Natalia Alonso / Pereira, Jonatas Luiz / da Silva, Israel Tojal / Defelicibus, Alexandre / Aoki, Mateus Nóbrega / Rivas, Javier De Las / Dos Reis, Rodolfo Borges / Zanette, Dalila Lucíola

Urologic oncology

2024 Band 42, Heft 3, Seite(n) 68.e11–68.e19

Abstract: Background: The median age for Prostate Cancer (PCa) diagnosis is 66 years, but 10% are diagnosed before 55 years. Studies on early-onset PCa remain both limited and controversial. This investigation sought to identify and characterize germline variants ...

Abstract	Background: The median age for Prostate Cancer (PCa) diagnosis is 66 years, but 10% are diagnosed before 55 years. Studies on early-onset PCa remain both limited and controversial. This investigation sought to identify and characterize germline variants within Brazilian PCa patients classified as either early or later onset disease. Methods: Peripheral blood DNA from 71 PCa patients: 18 younger (≤ 55 years) and 53 older (≥ 60 years) was used for Targeted DNA sequencing of 20 genes linked to DNA damage response, transcriptional regulation, cell cycle, and epigenetic control. Subsequent genetic variant identification was performed and variant functional impacts were analyzed with in silico prediction. Results: A higher frequency of variants in the BRCA2 and KMT2C genes across both age groups. KMT2C has been linked to the epigenetic dysregulation observed during disease progression in PCa. We present the first instance of KMT2C mutation within the blood of Brazilian PCa patients. Furthermore, out of the recognized variants within the KMT2C gene, 7 were designated as deleterious. Thirteen deleterious variants were exclusively detected in the younger group, while the older group exhibited 37 variants. Within these findings, 4 novel variants emerged, including 1 designated as pathogenic. Conclusions: Our findings contribute to a deeper understanding of the genetic factors associated with PCa susceptibility in different age groups, especially among the Brazilian population. This is the first investigation to explore germline variants specifically in younger Brazilian PCa patients, with high relevance given the genetic diversity of the population in Brazil. Additionally, our work presents evidence of functionally deleterious germline variants within the KMT2C gene among Brazilian PCa patients. The identification of novel and functionally significant variants in the KMT2C gene emphasizes its potential role in PCa development and warrants further investigation.
Mesh-Begriff(e)	Male ; Humans ; Aged ; Brazil ; Prostatic Neoplasms/pathology ; Germ-Line Mutation ; Mutation ; Germ Cells/pathology ; Genetic Predisposition to Disease
Sprache	Englisch
Erscheinungsdatum	2024-02-03
Erscheinungsland	United States
Dokumenttyp	Journal Article
ZDB-ID	1336505-8
ISSN	1873-2496 ; 1078-1439
ISSN (online)	1873-2496
ISSN	1078-1439
DOI	10.1016/j.urolonc.2024.01.015
Datenquelle	MEDical Literature Analysis and Retrieval System OnLINE

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Artikel ; Online: Screening of Exosome-Derived Proteins and Their Potential as Biomarkers in Diagnostic and Prognostic for Pancreatic Cancer.

Marin, Anelis Maria / Batista, Michel / Korte de Azevedo, Alexandre Luiz / Bombardelli Gomig, Talita Helen / Soares Caldeira Brant, Rodrigo / Chammas, Roger / Uno, Miyuki / Dias Araújo, Diogo / Zanette, Dalila Luciola / Nóbrega Aoki, Mateus

International journal of molecular sciences

2023 Band 24, Heft 16

Abstract: In the oncological area, pancreatic cancer is one of the most lethal diseases, with 5-year survival rising just 10% in high-development countries. This disease is genetically characterized ... ...

Abstract	In the oncological area, pancreatic cancer is one of the most lethal diseases, with 5-year survival rising just 10% in high-development countries. This disease is genetically characterized by
Mesh-Begriff(e)	Humans ; Early Detection of Cancer ; Prognosis ; Pancreatic Neoplasms/diagnosis ; Antineoplastic Combined Chemotherapy Protocols ; Exosomes ; Pancreatic Intraductal Neoplasms ; Proteomics ; Tandem Mass Spectrometry ; CA-19-9 Antigen ; Pancreatic Neoplasms
Chemische Substanzen	CA-19-9 Antigen
Sprache	Englisch
Erscheinungsdatum	2023-08-09
Erscheinungsland	Switzerland
Dokumenttyp	Journal Article
ZDB-ID	2019364-6
ISSN	1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online)	1422-0067
ISSN	1422-0067 ; 1661-6596
DOI	10.3390/ijms241612604
Datenquelle	MEDical Literature Analysis and Retrieval System OnLINE

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Artikel ; Online: Sample Preparation for Lab-on-a-Chip Systems in Molecular Diagnosis: A Review.

Cunha, Mylena Lemes / da Silva, Stella Schuster / Stracke, Mateus Cassaboni / Zanette, Dalila Luciola / Aoki, Mateus Nóbrega / Blanes, Lucas

Analytical chemistry

2021 Band 94, Heft 1, Seite(n) 41–58

Abstract	Rapid and low-cost molecular analysis is especially required for early and specific diagnostics, quick decision-making, and sparing patients from unnecessary tests and hospitals from extra costs. One way to achieve this objective is through automated molecular diagnostic devices. Thus, sample-to-answer microfluidic devices are emerging with the promise of delivering a complete molecular diagnosis system that includes nucleic acid extraction, amplification, and detection steps in a single device. The biggest issue in such equipment is the extraction process, which is normally laborious and time-consuming but extremely important for sensitive and specific detection. Therefore, this Review focuses on automated or semiautomated extraction methodologies used in lab-on-a-chip devices. More than 15 different extraction methods developed over the past 10 years have been analyzed in terms of their advantages and disadvantages to improve extraction procedures in future studies. Herein, we are able to explain the high applicability of the extraction methodologies due to the large variety of samples in which different techniques were employed, showing that their applications are not limited to medical diagnosis. Moreover, we are able to conclude that further research in the field would be beneficial because the methodologies presented can be affordable, portable, time efficient, and easily manipulated, all of which are strong qualities for point-of-care technologies.
Mesh-Begriff(e)	Humans ; Lab-On-A-Chip Devices ; Nucleic Acid Amplification Techniques ; Nucleic Acids ; Pathology, Molecular ; Point-of-Care Systems ; Specimen Handling
Chemische Substanzen	Nucleic Acids
Sprache	Englisch
Erscheinungsdatum	2021-12-06
Erscheinungsland	United States
Dokumenttyp	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	1508-8
ISSN	1520-6882 ; 0003-2700
ISSN (online)	1520-6882
ISSN	0003-2700
DOI	10.1021/acs.analchem.1c04460
Datenquelle	MEDical Literature Analysis and Retrieval System OnLINE

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