Artykuły w czasopismach: „Non-cellular diagnostics”

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Matulić, Maja, Paula Gršković, Andreja Petrović, Valerija Begić, Suzana Harabajsa i Petra Korać. "miRNA in Molecular Diagnostics". Bioengineering 9, nr 9 (9.09.2022): 459. http://dx.doi.org/10.3390/bioengineering9090459.

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MicroRNAs are a class of small non-coding RNA molecules that regulate gene expression on post-transcriptional level. Their biogenesis consists of a complex series of sequential processes, and they regulate expression of many genes involved in all cellular processes. Their function is essential for maintaining the homeostasis of a single cell; therefore, their aberrant expression contributes to development and progression of many diseases, especially malignant tumors and viral infections. Moreover, they can be associated with certain states of a specific disease, obtained in the least invasive manner for patients and analyzed with basic molecular methods used in clinical laboratories. Because of this, they have a promising potential to become very useful biomarkers and potential tools in personalized medicine approaches. In this review, miRNAs biogenesis, significance in cancer and infectious diseases, and current available test and methods for their detection are summarized.

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LIU, TIMON CHENG-YI, LING ZHU i XIANG-BO YANG. "PHOTOBIOMODULATION-MEDIATED PATHWAY DIAGNOSTICS". Journal of Innovative Optical Health Sciences 06, nr 01 (styczeń 2013): 1330001. http://dx.doi.org/10.1142/s1793545813300012.

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Cellular pathways are ordinarily diagnosed with pathway inhibitors, related gene regulation, or fluorescent protein markers. They are also suggested to be diagnosed with pathway activation modulation of photobiomodulation (PBM) in this paper. A PBM on a biosystem function depends on whether the biosystem is in its function-specific homeostasis (FSH). An FSH, a negative feedback response for the function to be performed perfectly, is maintained by its FSH-essential subfunctions and its FSH-non-essential subfunctions (FNSs). A function in its FSH or far from its FSH is called a normal or dysfunctional function. A direct PBM may self-adaptatively modulate a dysfunctional function until it is normal so that it can be used to discover the optimum pathways for an FSH to be established. An indirect PBM may self-adaptatively modulate a dysfunctional FNS of a normal function until the FNS is normal, and the normal function is then upgraded so that it can be used to discover the redundant pathways for a normal function to be upgraded.

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Gilmutdinova, Ilmira R., Irina S. Kudryashova, Elena Yu Kostromina, Maksim Yu Yakovlev, Inessa Kh Yafarova, Rinat G. Gilmutdinov, Irina A. Kaverina, Anton V. Ershov, Andrey N. Isaev i Alexey A. Moskalev. "Modern Approaches to Diagnostics and Correction of Aging Biomarkers". Bulletin of Rehabilitation Medicine 20, nr 6 (21.12.2021): 96–102. http://dx.doi.org/10.38025/2078-1962-2021-20-6-96-102.

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From the biomedicine point of, view ageing is a natural process, characterized by a gradual decrease in the physiological integrity and adaptive abilities of the body, leading to a violation of its functions and an increase in the risk of death with age. Demographic aging of the population is a serious socio-economic problem, both in Russia and around the world. The main cellular and molecular signs of aging include genome instability, telomere shortening, epigenetic alterations, impaired proteostasis, impaired nutrient recognition, mitochondrial dysfunction, cellular aging, the stem cell pool depletion and changes in intercellular interaction, extracellular matrix rigidity, as well as activation of retrotransposons and chronic inflammation. For these reasons, in modern healthcare, preventing premature aging and treating age-related diseases is becoming a priority task. This review presents modern approaches to the quantitative assessment of the aging process using aging biomarkers as functional parameters reflecting the biological organism age at the molecular, cellular, and organismal levels. This work also considers the actual non-drug and drug interventions allowing to slow down the development of age-associated pathological processes, allowing you to increase the quality and duration of life.

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Grigoruk, O. G., T. A. Moskvina, D. A. Tsoy, A. S. Stepanova, L. M. Bazulina, E. S. Sigitova, T. V. Ponomareva i in. "Endocervical adenocarcinomas. Cytological, histological, and molecular diagnostics". Tumors of female reproductive system 18, nr 2 (19.09.2022): 109–18. http://dx.doi.org/10.17650/1994-4098-2022-18-2-109-118.

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This study was undertaken to analyze the effectiveness of cytological diagnostics of endocervical adenocarcinomas. We compared conventional liquid-based cytology, histology, immunohistochemistry, and molecular testing. A total of 25 endocervical adenocarcinomas, including endocervical adenocarcinomas in situ, were diagnosed using cytological methods over a year. Liquid-based cytology ensured better detection of glandular differentiation signs than conventional cytology. After molecular testing for human papillomavirus (HPV), we performed retrospective analysis of cytological characteristics of all endocervical adenocarcinomas (n = 15).We identified specific cellular characteristics of HPV-associated typical and mucinous adenocarcinomas. We also observed 1 case of non-HPV-related clear-cell and 1 case of non-HPV-related mesonephral adenocarcinoma.Our findings suggest that endocervical adenocarcinomas are a heterogeneous group of tumors. Endocervical adenocarcinomas accounted for 10.7 % of all primary cervical carcinomas (n = 214). Eighty percent of all endocervical adenocarcinomas were HPV-related, whereas the remaining 20 % were HPV-negative. We found no cytological differences between invasive endocervical adenocarcinomas and adenocarcinomas in situ.Mutations detected in some of the patients are an important diagnostic criterion that specifies whether the tumor is rare.

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Solevåg, Anne Lee, Svetlana N. Zykova, Per Medbøe Thorsby i Georg M. Schmölzer. "Metabolomics to Diagnose Oxidative Stress in Perinatal Asphyxia: Towards a Non-Invasive Approach". Antioxidants 10, nr 11 (2.11.2021): 1753. http://dx.doi.org/10.3390/antiox10111753.

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There is a need for feasible and non-invasive diagnostics in perinatal asphyxia. Metabolomics is the study of small molecular weight products of cellular metabolism that may, directly and indirectly, reflect the level of oxidative stress. Saliva analysis is a novel approach that has a yet unexplored potential in metabolomics in perinatal asphyxia. The aim of this review was to give an overview of metabolomics studies of oxidative stress in perinatal asphyxia, particularly searching for studies analyzing non-invasively collected biofluids including saliva. We searched the databases PubMed/Medline and included 11 original human and 4 animal studies. In perinatal asphyxia, whole blood, plasma, and urine are the most frequently used biofluids used for metabolomics analyses. Although changes in oxidative stress-related salivary metabolites have been reported in adults, the utility of this approach in perinatal asphyxia has not yet been explored. Human and animal studies indicate that, in addition to antioxidant enzymes, succinate and hypoxanthine, as well acylcarnitines may have discriminatory diagnostic and prognostic properties in perinatal asphyxia. Researchers may utilize the accumulating evidence of discriminatory metabolic patterns in perinatal asphyxia to develop bedside methods to measure oxidative stress metabolites in perinatal asphyxia. Although only supported by indirect evidence, saliva might be a candidate biofluid for such point-of-care diagnostics.

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Hauptman, Nina, i Damjan Glavac. "MicroRNAs and long non-coding RNAs: prospects in diagnostics and therapy of cancer". Radiology and Oncology 47, nr 4 (1.12.2013): 311–18. http://dx.doi.org/10.2478/raon-2013-0062.

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AbstractBackground. Non-coding RNAs (ncRNAs) are key regulatory molecules in cellular processes, and are potential biomarkers in many diseases. Currently, microRNAs and long non-coding RNAs are being pursued as diagnostic and prognostic biomarkers, and as therapeutic tools in cancer, since their expression profiling is able to distinguish different cancer types and classify their sub-types.Conclusions. There are numerous studies confirming involvement of ncRNAs in cancer initiation, development and progression, but have only been recently identified as new diagnostic and prognostic tools. This can be beneficial in future medical cancer treatment options, since ncRNAs are natural antisense interactors included in regulation of many genes connected to survival and proliferation. Research is directed in development of useful markers for diagnosis and prognosis in cancer and in developing new RNA-based cancer therapies, of which some are already in clinical trials.

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Hoang, Johnson, Pooria Tajalli, Mina Omidiyan, Maria D. Marquez, Orawan Khantamat, Wirote Tuntiwechapikul, Chien-Hung Li i in. "Self-Assembled Monolayers Derived from Positively Charged Adsorbates on Plasmonic Substrates for MicroRNA Delivery: A Review". Journal of Nanotheranostics 4, nr 2 (8.05.2023): 171–200. http://dx.doi.org/10.3390/jnt4020009.

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MicroRNA (miRNA) has emerged as a promising alternative therapeutic treatment for cancer, but its delivery has been hindered by low cellular uptake and degradation during circulation. In this review, we discuss the various methods of delivering miRNA, including viral and non-viral delivery systems such as liposomes and nanoparticles. We also examine the use of nanoparticles for miRNA-based diagnostics. We focus specifically on non-viral delivery systems utilizing coinage metals in the form of nanoparticles and the use of self-assembled monolayers (SAMs) as a method of surface modification. We review the use of SAMs for the conjugation and delivery of small noncoding ribonucleic acid (ncRNA), particularly SAMs derived from positively charged adsorbates to generate charged surfaces that can interact electrostatically with negatively charged miRNA. We also discuss the effects of the cellular uptake of gold and other plasmonic nanoparticles, as well as the challenges associated with the degradation of oligonucleotides. Our review highlights the potential of SAM-based systems as versatile and robust tools for delivering miRNA and other RNAs in vitro and in vivo and the need for further research to address the challenges associated with miRNA delivery and diagnostics.

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Parekh, Palak R., Gregory M. Botting, Denise B. Thurber, Marika Boruszczak, William Murphy i Greg P. Bertenshaw. "Predictive biomarkers for response to trametinib in non-small cell lung cancer". Tumor Biology 44, nr 1 (9.12.2022): 249–67. http://dx.doi.org/10.3233/tub-220009.

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BACKGROUND: Non-small cell lung cancer (NSCLC) is a leading cause of cancer deaths. Current companion diagnostics use driver mutation sequencing to select patients for molecularly targeted agents (MTA), even though most patients lack actionable mutations. These diagnostics utilize static biomarkers, ignoring real-time tumor cell biology. OBJECTIVE: Trametinib is FDA-approved in combination with dabrafenib for BRAF V600E-positive NSCLC, however, it has plausible utility beyond these patients. We sought to identify novel biomarkers for maximizing trametinib application. METHODS: Trametinib responses were evaluated in 12 EGFR/BRAF wild-type (WT) NSCLC cell lines with diverse RAS mutational status. We identified three response categories by colony assay. Trametinib-induced molecular dynamics were studied using immunoassays and apoptosis/necrosis assays, to identify predictive response biomarkers. RESULTS: p27 accumulation and cyclin D1 downregulation suggested universal cell cycle arrest with trametinib. However, 4 cell lines showed PARP cleavage and 8 showed increased phospho-4E-BP1, suggesting varied cellular outcomes from apoptosis, necrosis, senescence to autophagy. Cleaved PARP, phospho-4E-BP1 and phospho-AKT expression can predict these outcomes. CONCLUSIONS: Trametinib monotherapy outcome may depend upon cellular context more than oncogenic mutation status. In BRAF WT NSCLC, trametinib may be best suited for combination therapy and dynamic biomarkers could select combinations and predict responses.

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Shiv Swaroop i Thangminlal Vaiphei S. "Potential Roles of Long Non-Coding RNAs (lncRNAs) in Stress Response Regulation". International Journal of Research in Pharmaceutical Sciences 11, SPL4 (21.12.2020): 2385–89. http://dx.doi.org/10.26452/ijrps.v11ispl4.4482.

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The non-coding RNAs (ncRNAs) are functional RNA transcripts involved in gene regulation at the level of transcription and post-transcription. There have been vital pieces of evidence to support the role of regulatory non-coding RNAs in the eukaryotic genome in recent years. The ncRNAs are also associated with post-translational modifications such as histone modification, heterochromatin formation, DNA methylation and other key molecules which are involved in regulating chromatin structures for gene expression. LncRNAs (long non-coding RNAs) are the most diverse, biologically active transcripts without significant open reading frames (ORFs) and represent the majority of ncRNAs populations in the human genome. Emerging pieces of evidence suggest the role of ncRNAs in a wide range of human diseases, including cardiovascular, Alzheimer, and cancer. Several reports in the recent past also supported their involvement in the modulation of various cellular responses, although the mechanisms of ncRNAs mediated gene regulations are still not fully understood. This review paper highlights the importance of lncRNAs in cellular stress response such as DNA damaging ionizing radiation that will encourage research in thrust areas of therapeutics and diagnostics. The involvement of important lncRNAs in regulating biological processes, responses to ionizing and non-ionizing radiation, as well as methods for the analysis of their cellular expression has been discussed.

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Gubenko, Marina S., Vitaliy I. Loginov, Aleksey M. Burdennyy, Irina V. Pronina, Svetlana V. Khokhlova i Sergey S. Pertsov. "Role of Microribonucleic acid in the Carcinogenesis of Non-Small-Cell Lung Cancer". I.P. Pavlov Russian Medical Biological Herald 30, nr 1 (31.03.2022): 123–32. http://dx.doi.org/10.17816/pavlovj71395.

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INTRODUCTION: Lung cancer is the most common malignant neoplasm. Despite advances in target therapy, immunotherapy, and chemotherapy, non-small cell lung cancer remains the major cause of cancer-related death worldwide. Tumor development is a complex process that depends on the influence of environmental factors and genetic predisposition. Although oncogenic factors have received much attention, the main mechanisms for oncogenesis are still poorly understood. Thus, studying the oncogenic mechanisms, including those with the involvement of microribonucleic acid (microRNA), is important for the diagnostics and treatment of malignant neoplasms. MicroRNA (miRNA) belong to the class of small non-coding ribonucleic acids that are involved in various cellular biological processes, including epithelialmesenchymal transition, apoptosis, proliferation, invasion, and metastatic dissemination of cancer cells. Recent publications show that the course of the oncological disease can be predicted by evaluating the expressions of some miRNAs. Therefore, miRNAs serve as promising diagnostic and therapeutic targets in oncological diseases. CONCLUSION: This review summarizes data on the role in carcinogenesis and prognostic significance of several miRNA (i.e., miRNA-128, -4500, -222, -224, -124, -125b, -127, -129-2, -137, and -375) in non-small cell lung cancer.

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Thomas, Rebecca, i Mark Chambers. "Review of Methods Used for Diagnosing Tuberculosis in Captive and Free-Ranging Non-Bovid Species (2012–2020)". Pathogens 10, nr 5 (11.05.2021): 584. http://dx.doi.org/10.3390/pathogens10050584.

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The Mycobacterium tuberculosis complex (MTBC) is a group of bacteria that cause tuberculosis (TB) in diverse hosts, including captive and free-ranging wildlife species. There is significant research interest in developing immunodiagnostic tests for TB that are both rapid and reliable, to underpin disease surveillance and control. The aim of this study was to carry out an updated review of diagnostics for TB in non-bovid species with a focus predominantly on those based on measurement of immunity. A search was carried out to identify relevant papers meeting a pre-defined set of inclusion criteria. Forty-one papers were identified from this search, from which only twenty papers contained data to measure and compare diagnostic performance using diagnostic odds ratio. The diagnostic tests from each study were ranked based on sensitivity, specificity, and diagnostic odds ratio to define high performing tests. High sensitivity and specificity values across a range of species were reported for a new antigenic target, P22 complex, demonstrating it to be a reliable and accurate antigenic target. Since the last review of this kind was undertaken, the immunodiagnosis of TB in meerkats and African wild dogs was reported for the first time. Suid species showed the most consistent immunological responses and highlight a potential dichotomy between humoral and cellular immune responses.

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Pereira, Jorge A. M., Priscilla Porto-Figueira, Ravindra Taware, Pritam Sukul, Srikanth Rapole i José S. Câmara. "Unravelling the Potential of Salivary Volatile Metabolites in Oral Diseases. A Review". Molecules 25, nr 13 (7.07.2020): 3098. http://dx.doi.org/10.3390/molecules25133098.

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Fostered by the advances in the instrumental and analytical fields, in recent years the analysis of volatile organic compounds (VOCs) has emerged as a new frontier in medical diagnostics. VOCs analysis is a non-invasive, rapid and inexpensive strategy with promising potential in clinical diagnostic procedures. Since cellular metabolism is altered by diseases, the resulting metabolic effects on VOCs may serve as biomarkers for any given pathophysiologic condition. Human VOCs are released from biomatrices such as saliva, urine, skin emanations and exhaled breath and are derived from many metabolic pathways. In this review, the potential of VOCs present in saliva will be explored as a monitoring tool for several oral diseases, including gingivitis and periodontal disease, dental caries, and oral cancer. Moreover, the analytical state-of-the-art for salivary volatomics, e.g., the most common extraction techniques along with the current challenges and future perspectives will be addressed unequivocally.

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Носик, А. В., С. В. Коротков, В. В. Смольникова, В. Ю. Гриневич, М. В. Дмитриева, И. И. Пикиреня, О. В. Калачик, С. И. Кривенко, А. Е. Щерба i О. О. Руммо. "Clinical Effectiveness of Minimally Invasive Diagnostics of Cellular Rejection after Kidney Transplantation". Хирургия. Восточная Европа, nr 1-2 (18.05.2020): 24–37. http://dx.doi.org/10.34883/pi.2020.9.1.014.

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Цель. Продемонстрировать клиническую эффективность применения разработанного алгоритма диагностики клеточного отторжения почечного аллографта в отдаленные сроки после трансплантации. Материалы и методы. Проведено проспективное интервенционное исследование, включившее 10 реципиентов трансплантата почки. В основную группу исследования включены 5 пациентов с отторжением почечного аллографта, диагностированным разработанным алгоритмом. Контрольную группу составили 5 пациентов с неиммунной дисфункцией трансплантата. В основной группе исследования начата специфическая терапия отторжения по результатам предлагаемого теста. Всем участникам произведена биопсия трансплантата почки. Результаты и обсуждение. Применение специфической терапии по результатам предлагаемого алгоритма привело к снижению сывороточного креатинина (140 127 111 мкмоль/л) и росту скорости клубочковой фильтрации (43,5 46,5 58,6 мл/мин). Сравнение результатов предлагаемого метода с результатами гистологического исследования продемонстрировало хорошие диагностические характеристики. Чувствительность и специфичность были равны 100,0 (95 ДИ 45,97100,0) для обеих характеристик. Выводы. Применение специфической терапии позднего клеточного отторжения по результатам предлагаемого алгоритма является эффективным. Предлагаемый алгоритм обладает хорошими диагностическими характеристиками в сравнении с гистологическим исследованием. Purpose. To demonstrate clinical effectiveness of use of the developed algorithm for diagnostics of cell rejection of renal allograft in the long term after transplantation. Materials and methods. We conducted a prospective interventional comparative study, which included 10 patients. The experimental study group included 5 patients with rejection, which was diagnosed with the developed algorithm. The control group consisted of 5 patients with non- immune transplant dysfunction. In the experimental study group, specic rejection therapy was initiated, according to the results of the proposed test. All the participants underwent a kidney transplant biopsy. Results and discussion. The use of specic therapy based on the results of the proposed algorithm led to decrease of serum creatinine (140 127 111 mol/l) and increase of GRF (43.5 46.5 58.6 ml / min). The comparison of the results of the proposed method with the results of the histological study demonstrated good diagnostic characteristics. Sensitivity and specicity were 100.0 (95 CI 45.97100.0) for both characteristics. Conclusions. The use of specic therapy for treatment of late cellular rejection based on the results of the proposed algorithm is effective. The proposed algorithm has good diagnostic characteristics in comparison with histological examination

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Jothimani, Ganesan, Sushmitha Sriramulu, Yashna Chabria, Xiao-Feng Sun, Antara Banerjee i Surajit Pathak. "A Review on Theragnostic Applications of Micrornas and Long Non- Coding RNAs in Colorectal Cancer". Current Topics in Medicinal Chemistry 18, nr 30 (13.02.2019): 2614–29. http://dx.doi.org/10.2174/1568026619666181221165344.

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Colorectal cancer (CRC) is a heterogeneous malignancy leading to increased mortality and poor prognosis due to the lack of efficient early diagnostics. Metastasis of the tumor being the most common cause of mortality is accountable for almost 90% of CRC associated deaths. Intensified screening procedures and molecular target identification has inflated the median survival rate of in CRC patients. microRNAs (miRNAs) and long non-coding RNAs (lncRNAs) have come forward as potential targets for developing a novel approach in CRC theragnostics. Non-coding RNA (ncRNAs) sequences are abundantly present and thereby play a vital role in several biological processes such as cellular organization, cell fate determination, proliferation, apoptosis, tissue homeostasis maintenance as well as pathological conditions such as cancer by acting as post transcriptional regulators of gene expression. Several studies have highlighted the involvement of these ncRNAs in CRC development. However, the molecular mechanism involved in regulating CRC has not been clearly elucidated. This review, throws light upon the several non-coding RNAs involved in CRC with a focus on novel mechanisms of action, recent advances in the regulatory mechanisms that control the gene expression related to carcinogenesis. Furthermore, the potential role of ncRNAs as diagnostic as well as therapeutic targets has been reviewed.

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Duetz, Carolien, Sofie Van Gassen, Theresia M. Westers, Florentien in t Hout, Eline Cremers, Canan Alhan, Costa Bachas i in. "Machine Learning-Based Flow Cytometry Diagnostics in Myelodysplastic Syndromes: Validation in the HOVON89 Clinical Trial (EudraCT 2008-002195-10)". Blood 136, Supplement 1 (5.11.2020): 10–12. http://dx.doi.org/10.1182/blood-2020-136719.

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Introduction Flow cytometry is a recommended tool in the diagnostic work-up of cytopenic patients suspected for myelodysplastic syndromes. Currently used flow cytometry scores rely on human assessment of dysplastic features in the bone marrow. Although proven useful, these methods are labor intensive and require a high level of expertise. Therefore, we previously developed a machine learning-based workflow for flow cytometry diagnostics in MDS by combining computational cell detection and a machine learning-classifier. This workflow outperformed traditional diagnostic scores with respect to accuracy (sensitivity 85-97%, specificity 93-97%), time investment (<30 seconds) and required materials (manuscript submitted). In the present study, we validated sensitivity of the workflow in a well-characterized clinical trial cohort (HOVON89 EudraCT 2008-002195-10) of lower risk MDS patients. Method Patient inclusion and characteristics Very low to intermediate risk MDS patients enrolled in the HOVON89 clinical trial (EudraCT 2008-002195-10) were included. 53 patients met the additional inclusion criteria, concerning written consent for add-on studies and availability of required flow cytometry data. Sample preparation Bone marrow samples were processed for flow cytometry analysis according to the European Leukemia Net guidelines. This study focused on the antibody combination optimized for assessment of myeloid progenitors and erythroid dysplasia (CD45, CD34, CD117, HLA-DR, CD71, CD36, CD105, CD33, sideward light scatter (SSC) and forward light scatter (FSC)). Machine learning-based workflow The machine learning-based workflow was developed in a prior study based on a reference cohort consisting of MDS patients without excess of blasts(n=67) and non-MDS cases (n=81) (Figure 1). MDS patients were diagnosed based on (cyto)morphology, cytogenetics and clinical follow-up. Non-MDS cases were patients with confirmed non-neoplastic cytopenias (n=69) and age-matched healthy individuals (n=12). Results In the validation cohort, the machine learning-based diagnostic workflow classified 49 out of 53 patients correctly, reaching a sensitivity of 92%. The workflow outperformed two currently used diagnostic tools for MDS flow cytometry, the Ogata score and integrated flow cytometry score (iFS). The former obtained 72% sensitivity (McNemar: p = 0.001) and the latter 83% sensitivity (McNemar: p = 0.06) in the validation cohort. Per patient, time required for automated analysis was less than 30 seconds. All four MDS patients that classified false negatively had a normal karyotype and (very) low risk disease according to the IPSS-r. In three out of four patients, no mutations or MDS-associated immunophenotypic features were detected. One patients was diagnosed as MDS-MLD and three patients as MDS-RS-SLD according to the WHO 2016 classification. The ten most relevant cellular features that discriminated between MDS and non-MDS patients in the reference data were confirmed in the current validation cohort. All ten features of MDS patients in the validation cohort were significantly different from non-MDS patients of the reference cohort (all features, p < 0.00001) (Figure 2). Seven out of ten features were similar in MDS patients of the validation cohort compared to those of the MDS patients of the reference cohort (p>0.05) (Figure 2). Conclusion In this validation study, we confirmed accuracy of machine learning-based flow cytometry diagnostics in lower risk MDS. The workflow obtained 92% sensitivity, which is in accordance with results from our previous study (85-97%), and outperformed currently used diagnostic flow cytometry scores for MDS (i.e. Ogata score and iFS). In our previous study specificity was 95% in both reference and test cohorts. Cellular features, most discriminative for diagnosis, were confirmed in the validation cohort, emphasizing robustness of the method. Additional benefits of this approach are the reduction in analysis time to less than thirty seconds per patient, reduction of required antibodies and increased reproducibility. Disclosures van de Loosdrecht: celgene: Honoraria; novartis: Honoraria.

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K. Tapasya, K. Tapasya, Ashmitha Suresh Kumar, Arunasalam Dharmarajan i Venkatachalam Deepa Parvathi. "Nanocarriers: The Promising Future to Cancer Diagnostics and Treatment". Biomedical and Pharmacology Journal 15, nr 2 (30.06.2022): 785–802. http://dx.doi.org/10.13005/bpj/2416.

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Nanotechnology recently gained attention for the novel and successful tools it has thus far provided for cancer diagnosis and treatment. Some of them include lipid-based carriers such as liposomes and metal-based particles such as nanoshells (NSs), used for anti-cancer drug delivery for the most part. Each one of these systems has been carefully designed in order to bypass the obstacles brought forward by conventional diagnosis and treatment strategies. These challenges include non-specificity, premature drug release and toxicity. From research conducted over the years it is clear that nanocarriers ameliorate bioavailability, specificity and accumulation of the drugs at the target site. These improvements can be explained by their easily adjustable physical and chemical properties. Alterations to their size and surface structure are often made to enhance their accumulation at the target sites and overall targeting capabilities respectively. Some nanocarriers such as quantum dots (QDs) and carbon nanotubes (CNTs) display excellent fluorescent properties and are useful candidates for imaging techniques and fluorescence-guided surgery. Another group of promising nanoparticles is biomimetic nanoparticles that mimic the functionality of biological components. These NPs are designed to mimic basic cellular and physical features of the source cells and their surface. This type of NPs construct is exploited for its unique characteristics that aid in effective interaction with complex biological systems, consequently enhancing therapeutic outcomes After establishing them as adequate tools for drug delivery and imaging, nanocarriers are now being tested in combined cancer treatment strategies. This review provides an understanding of the salient nano-devices and their applications in oncology.

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Li, Yiwei, Mohammed Najeeb Al Hallak, Philip A. Philip, Asfar S. Azmi i Ramzi M. Mohammad. "Non-Coding RNAs in Pancreatic Cancer Diagnostics and Therapy: Focus on lncRNAs, circRNAs, and piRNAs". Cancers 13, nr 16 (19.08.2021): 4161. http://dx.doi.org/10.3390/cancers13164161.

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Pancreatic cancer is an aggressive malignance with high mortality. The lack of early diagnosis and effective therapy contributes to the high mortality of this deadly disease. For a long time being, the alterations in coding RNAs have been considered as major targets for diagnosis and treatment of pancreatic cancer. However, with the advances in high-throughput next generation of sequencing more alterations in non-coding RNAs (ncRNAs) have been discovered in different cancers. Further mechanistic studies have demonstrated that ncRNAs such as long noncoding RNAs (lncRNA), circular RNAs (circRNA) and piwi-interacting RNA (piRNA) play vital roles in the regulation of tumorigenesis, tumor progression and prognosis. In recent years, increasing studies have focused on the roles of ncRNAs in the development and progression of pancreatic cancer. Novel findings have demonstrated that lncRNA, circRNA, and piRNA are critically involved in the regulation of gene expression and cellular signal transduction in pancreatic cancer. In this review, we summarize the current knowledge of roles of lncRNA, circRNA, and piRNA in the diagnosis and prognosis of pancreatic cancer, and molecular mechanisms underlying the regulation of these ncRNAs and related signaling in pancreatic cancer therapy. The information provided here will help to find new strategies for better treatment of pancreatic cancer.

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Zhang, Zhao, Xiaowen Huang, Ke Liu, Tiancong Lan, Zixin Wang i Zhen Zhu. "Recent Advances in Electrical Impedance Sensing Technology for Single-Cell Analysis". Biosensors 11, nr 11 (22.11.2021): 470. http://dx.doi.org/10.3390/bios11110470.

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Cellular heterogeneity is of significance in cell-based assays for life science, biomedicine and clinical diagnostics. Electrical impedance sensing technology has become a powerful tool, allowing for rapid, non-invasive, and label-free acquisition of electrical parameters of single cells. These electrical parameters, i.e., equivalent cell resistance, membrane capacitance and cytoplasm conductivity, are closely related to cellular biophysical properties and dynamic activities, such as size, morphology, membrane intactness, growth state, and proliferation. This review summarizes basic principles, analytical models and design concepts of single-cell impedance sensing devices, including impedance flow cytometry (IFC) to detect flow-through single cells and electrical impedance spectroscopy (EIS) to monitor immobilized single cells. Then, recent advances of both electrical impedance sensing systems applied in cell recognition, cell counting, viability detection, phenotypic assay, cell screening, and other cell detection are presented. Finally, prospects of impedance sensing technology in single-cell analysis are discussed.

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Pisarska, Justyna, i Katarzyna Baldy-Chudzik. "MicroRNA-Based Fingerprinting of Cervical Lesions and Cancer". Journal of Clinical Medicine 9, nr 11 (15.11.2020): 3668. http://dx.doi.org/10.3390/jcm9113668.

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The regulatory functions of microRNA (miRNA) are involved in all processes contributing to carcinogenesis and response to viral infections. Cervical cancer in most cases is caused by the persistence of high-risk human papillomavirus (HR-HPV) infection. While oncogenic human papillomaviruses induce aberrant expression of many cellular miRNAs, this dysregulation could be harnessed as a marker in early diagnosis of HR-HPV infection, cervical squamous intraepithelial lesions, and cancer. In recent years, growing data indicate that miRNAs show specific patterns at various stages of cervical pathology. The aim of this review is to systematize current reports on miRNA capacity that can be utilized in personalized diagnostics of cervical precancerous and cancerous lesions. The analysis of the resources available in online databases (National Center for Biotechnology Information—NCBI, PubMed, ScienceDirect, Scopus) was performed. To date, no standardized diagnostic algorithm using the miRNA pattern in cervical pathology has been defined. However, the high sensitivity and specificity of the reported assays gives hope for the development of non-invasive diagnostic tests that take into account the heterogeneity of tumor-related changes. Due to this variability resulting in difficult to predict clinical outcomes, precise molecular tools are needed to improve the diagnostic and therapeutic process.

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Masters, Thao, Aditya Bhagwate, Mrunal Dehankar, Kerryl Greenwood-Quaintance, Matthew P. Abdel, Robin Patel i Robin Patel. "1193. Human Transcriptomic Analysis of Periprosthetic Joint Infection". Open Forum Infectious Diseases 7, Supplement_1 (1.10.2020): S619. http://dx.doi.org/10.1093/ofid/ofaa439.1378.

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Abstract Background Periprosthetic joint infection (PJI), a devastating complication of total joint replacement, is of incompletely understood pathogenesis and may sometimes be challenging to clinically distinguish from other causes of arthroplasty failure. Methods We characterized human gene expression in 93 specimens derived from surfaces of resected arthroplasties, comparing transcriptomes of subjects with infection- versus non-infection-associated arthroplasty failure. Results Differential gene expression analysis confirmed the association of 28 previously investigated biomarkers with PJI- bactericidal/permeability increasing protein (BPI), cathelicidin antimicrobial peptide (CAMP), chemokines CCL3, CCL4, and CXCL2, colony stimulating factor 2 receptor (CSF2RB), colony stimulating factor 3 (CSF3), alpha-defensin (DEFA4), receptor CD64B, intercellular adhesion molecule 1 (ICAM1), IFNG, IL13RA2, IL17D, IL1A, IL1B, IL1RN, IL2RA, IL2RG, IL5RA, IL6, IL8, lipopolysaccharide binding protein (LBP), lipocalin (LCN2), lactate dehydrogenase C (LDHC), lactotransferrin (LTF), matrix metallopeptidase 3 (MMP3), peptidase inhibitor 3 (PI3), and vascular endothelial growth factor A (VEGFA), as well as identified three novel molecules with diagnostic potential for detection of PJI- chemokine CCL20, coagulation factor VII (F7), B cell receptor FCRL4. Comparative analysis of infections caused by staphylococcal versus non-staphylococcal and Staphylococcus aureus versus Staphylococcus epidermidis showed significant elevated expression of IL13, IL17D, and metalloprotease protein MMP3 in staphylocococcal infections, and increased expression of IL1B, IL8, and platelet factor PF4V1 in S. aureus infections. Pathway analysis of over-presented genes suggested activation of host immune response and cellular maintenance and repair functions in response to invasion of infectious agents. Conclusion Our study provides new potential targets for diagnosis of PJI and targets for differentiation of PJI-associated infectious agents. Disclosures Matthew P. Abdel, MD, Dr. Abdel receives royalties from Stryker on certain hip and knee products, and is a paid consultant for Stryker. (Consultant) Robin Patel, MD, Accelerate Diagnostics (Grant/Research Support)CD Diagnostics (Grant/Research Support)Contrafect (Grant/Research Support)Curetis (Consultant)GenMark Diagnostics (Consultant)Heraeus Medical (Consultant)Hutchison Biofilm Medical Solutions (Grant/Research Support)Merck (Grant/Research Support)Next Gen Diagnostics (Consultant)PathoQuest (Consultant)Qvella (Consultant)Samsung (Other Financial or Material Support, Dr. Patel has a patent on Bordetella pertussis/parapertussis PCR issued, a patent on a device/method for sonication with royalties paid by Samsung to Mayo Clinic, and a patent on an anti-biofilm substance issued.)Selux Dx (Consultant)Shionogi (Grant/Research Support)Specific Technologies (Consultant)

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Balihodzic, Amar, Dominik A. Barth, Felix Prinz i Martin Pichler. "Involvement of Long Non-Coding RNAs in Glucose Metabolism in Cancer". Cancers 13, nr 5 (26.02.2021): 977. http://dx.doi.org/10.3390/cancers13050977.

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The rapid and uncontrolled proliferation of cancer cells is supported by metabolic reprogramming. Altered glucose metabolism supports cancer growth and progression. Compared with normal cells, cancer cells show increased glucose uptake, aerobic glycolysis and lactate production. Byproducts of adjusted glucose metabolism provide additional benefits supporting hallmark capabilities of cancer cells. Long non-coding RNAs (lncRNAs) are a heterogeneous group of transcripts of more than 200 nucleotides in length. They regulate numerous cellular processes, primarily through physical interaction with other molecules. Dysregulated lncRNAs are involved in all hallmarks of cancer including metabolic alterations. They may upregulate metabolic enzymes, modulate the expression of oncogenic or tumor-suppressive genes and disturb metabolic signaling pathways favoring cancer progression. Thus, lncRNAs are not only potential clinical biomarkers for cancer diagnostics and prediction but also possible therapeutic targets. This review summarizes the lncRNAs involved in cancer glucose metabolism and highlights their underlying molecular mechanisms.

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König, Karsten. "Two-Photon near Infrared Excitation in Living Cells". Journal of Near Infrared Spectroscopy 5, nr 1 (styczeń 1997): 27–34. http://dx.doi.org/10.1255/jnirs.97.

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Non-linear effects due to two-photon near infrared (NIR) excitation of endogenous and exogenous cellular chromophores allow novel techniques in tissue, cell and biomolecule diagnostics, as well as in intracellular micromanipulation (e.g. intracellular photochemistry). Two-photon NIR excitation may also result in cell damage effects. The high photon intensities (1024 photons cm−2 s−1) required for non-resonant two-photon excitation can be achieved by diffraction-limited focusing of continuous wave (cw) laser beams (cw microbeams) with powers in the mW range. For example, NIR traps (“laser tweezers”) used as force transducers and micromanipulation tools in cellular and molecular biology are sources of two-photon excitation. NIR traps can induce two-photon excited visible fluorescence and, in the case of <800 nm-traps, UVA-like cell damage. Multimode cw microbeams may enhance non-linear effects due to longitudinal mode-beating. To perform high scan rate two-photon fluorescence imaging, the application of ultrashort laser pulses of moderate peak power but low average power (pulsed microbeams) is required. In NIR femtosecond microscopes, non-destructive imaging of two-photon excited fluorophores in various human and culture cells was demonstrated for <2 mW average powers, <200 mW peak powers and 400 GW cm−2 intensities (700–800 nm, ∼150 fs, ∼100 MHz). However, higher average power levels may result in failed cell reproduction and cell death due to intracellular optical breakdown. In addition, destructive transient local heating and μN force generation may occur.

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Lehmann, Felix, Heiko Slanina, Martin Roderfeld, Elke Roeb, Jonel Trebicka, John Ziebuhr, Wolfram H. Gerlich, Christian G. Schüttler, Bernhard Schlevogt i Dieter Glebe. "A Novel Insertion in the Hepatitis B Virus Surface Protein Leading to Hyperglycosylation Causes Diagnostic and Immune Escape". Viruses 15, nr 4 (25.03.2023): 838. http://dx.doi.org/10.3390/v15040838.

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Chronic hepatitis B virus (HBV) infection is a global health threat. Mutations in the surface antigen of HBV (HBsAg) may alter its antigenicity, infectivity, and transmissibility. A patient positive for HBV DNA and detectable but low-level HBsAg in parallel with anti-HBs suggested the presence of immune and/or diagnostic escape variants. To support this hypothesis, serum-derived HBs gene sequences were amplified and cloned for sequencing, which revealed infection with exclusively non-wildtype HBV subgenotype (sgt) D3. Three distinct mutations in the antigenic loop of HBsAg that caused additional N-glycosylation were found in the variant sequences, including a previously undescribed six-nucleotide insertion. Cellular and secreted HBsAg was analyzed for N-glycosylation in Western blot after expression in human hepatoma cells. Secreted HBsAg was also subjected to four widely used, state-of-the-art diagnostic assays, which all failed to detect the hyperglycosylated insertion variant. Additionally, the recognition of mutant HBsAg by vaccine- and natural infection-induced anti-HBs antibodies was severely impaired. Taken together, these data suggest that the novel six-nucleotide insertion as well as two other previously described mutations causing hyperglycosylation in combination with immune escape mutations have a critical impact on in vitro diagnostics and likely increase the risk of breakthrough infection by evasion of vaccine-induced immunity.

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Bogomolov, A. I., I. L. Davydkin, E. A. Savinov, N. S. Popel'nyuk i K. V. Naumova. "Assessing risks of developing myeloproliferative diseases complications with laser doppler flowmetry". Health Risk Analysis, nr 3 (wrzesień 2020): 160–68. http://dx.doi.org/10.21668/health.risk/2020.3.19.

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Public healthcare in Russia faces many challenges; meeting them requires non-standard and innovative approaches. The set tasks are being solved within the «Public healthcare development» State program. A sub-program within it is called «Development and implementation of innovative diagnostics, prevention, and treatment procedures as well as basics of personified medicine». This sub-program involves wide use of information and digital technologies. Personified medicine envisages developing such methods that would allow early detection of a probable disease even at a preliminary stage in examining a patient; this detection is to be based on a simple and relatively cheap diagnostic technology and to provide a medical center with reliable data on detected signs of a disease for a further diagnosis. Mass use of such technologies also requires truly reliable mathematic procedures and models for putting a preliminary diagnosis. At present cardiovascular diseases are still the leading cause of death all over the world; they develop due to variable factors including influence exerted by malignant neoplasms and also due to chemotherapy. The paper contains data collected by contemporary medical experts on case histories and complications of myeloproliferative diseases caused by vascular system pathology that holds the first rank place as per mortality worldwide. It was detected that both pathological cellular mass and medications applied to treat myeloproliferative neoplasms could produce adverse effects on vascular endothelium damage to which plays the leading role in cardiovascular continuum. To assess risks of myeloproliferative diseases complications, we examined patients using Laser Doppler Flowmetry (LDF). The results were processed with a logistic regression model. As per ROC-analysis results the obtained diagnostic criterion has sensitivity (1 – β) and specificity, (1 – α) that are equal to 0.87 and 0.96 accordingly, and it means diagnostics is high-quality. The procedure and the mode can be applied in digital medicine.

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Bogomolov, A. I., I. L. Davydkin, E. A. Savinov, N. S. Popel'nyuk i K. V. Naumova. "Assessing risks of developing myeloproliferative diseases complications with laser doppler flowmetry". Health Risk Analysis, nr 3 (wrzesień 2020): 160–68. http://dx.doi.org/10.21668/health.risk/2020.3.19.eng.

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Public healthcare in Russia faces many challenges; meeting them requires non-standard and innovative approaches. The set tasks are being solved within the «Public healthcare development» State program. A sub-program within it is called «Development and implementation of innovative diagnostics, prevention, and treatment procedures as well as basics of personified medicine». This sub-program involves wide use of information and digital technologies. Personified medicine envisages developing such methods that would allow early detection of a probable disease even at a preliminary stage in examining a patient; this detection is to be based on a simple and relatively cheap diagnostic technology and to provide a medical center with reliable data on detected signs of a disease for a further diagnosis. Mass use of such technologies also requires truly reliable mathematic procedures and models for putting a preliminary diagnosis. At present cardiovascular diseases are still the leading cause of death all over the world; they develop due to variable factors including influence exerted by malignant neoplasms and also due to chemotherapy. The paper contains data collected by contemporary medical experts on case histories and complications of myeloproliferative diseases caused by vascular system pathology that holds the first rank place as per mortality worldwide. It was detected that both pathological cellular mass and medications applied to treat myeloproliferative neoplasms could produce adverse effects on vascular endothelium damage to which plays the leading role in cardiovascular continuum. To assess risks of myeloproliferative diseases complications, we examined patients using Laser Doppler Flowmetry (LDF). The results were processed with a logistic regression model. As per ROC-analysis results the obtained diagnostic criterion has sensitivity (1 – β) and specificity, (1 – α) that are equal to 0.87 and 0.96 accordingly, and it means diagnostics is high-quality. The procedure and the mode can be applied in digital medicine.

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Sulewska, Anetta, Lothar Pilz, Christian Manegold, Rodryg Ramlau, Radoslaw Charkiewicz i Jacek Niklinski. "A Systematic Review of Progress toward Unlocking the Power of Epigenetics in NSCLC: Latest Updates and Perspectives". Cells 12, nr 6 (15.03.2023): 905. http://dx.doi.org/10.3390/cells12060905.

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Epigenetic research has the potential to improve our understanding of the pathogenesis of cancer, specifically non-small-cell lung cancer, and support our efforts to personalize the management of the disease. Epigenetic alterations are expected to have relevance for early detection, diagnosis, outcome prediction, and tumor response to therapy. Additionally, epi-drugs as therapeutic modalities may lead to the recovery of genes delaying tumor growth, thus increasing survival rates, and may be effective against tumors without druggable mutations. Epigenetic changes involve DNA methylation, histone modifications, and the activity of non-coding RNAs, causing gene expression changes and their mutual interactions. This systematic review, based on 110 studies, gives a comprehensive overview of new perspectives on diagnostic (28 studies) and prognostic (25 studies) epigenetic biomarkers, as well as epigenetic treatment options (57 studies) for non-small-cell lung cancer. This paper outlines the crosstalk between epigenetic and genetic factors as well as elucidates clinical contexts including epigenetic treatments, such as dietary supplements and food additives, which serve as anti-carcinogenic compounds and regulators of cellular epigenetics and which are used to reduce toxicity. Furthermore, a future-oriented exploration of epigenetic studies in NSCLC is presented. The findings suggest that additional studies are necessary to comprehend the mechanisms of epigenetic changes and investigate biomarkers, response rates, and tailored combinations of treatments. In the future, epigenetics could have the potential to become an integral part of diagnostics, prognostics, and personalized treatment in NSCLC.

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Deryabina, S. S., D. A. Cheremokhin, I. A. Tuzankina, M. A. Bolkov i Kh Shinvary. "Retrospective analysis of pediatric primary immunodeficiencies with congenital heart defects". Russian Journal of Immunology 23, nr 4 (15.10.2020): 505–14. http://dx.doi.org/10.46235/1028-7221-492-ara.

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Around 20,000 children with heart defects are born annually in the Russian Federation, among which around 5,000 children are manifested by its critical course that requires surgical correction within the first days after birth. Although timely pre- and postnatal diagnostics of congenital heart pathology allows to provide proper therapeutic and surgical assistance for such patients, it is hard to minimize severe complications at any stage of the child’s treatment. Quite often, post-surgery patients are unable to cope with serious complications caused by infectious diseases of the respiratory tract, endocardium and meninges. A special place is taken by cases of severe postoperative period: long-term non-healing postoperative wounds and septic disorders leading to patient death. Here we put forward an idea that congenital heart disease may serve as a marker of primary immunodeficiency characterized by lowered parameters of cellular and humoral immunity. Current retrospective study included 29 children died within the first year of life, whose medical records demonstrated confirmed congenital heart disease and manifested signs of immune-dependent pathology. There were analyzed data regarding pregnancy course in order to identify factors contributing to early diagnostics of immune-related pathology associated with CHD. The analysis of existing problems affecting proper diagnostics and treatment of children with congenital heart disease coupled to “covert” PID form was performed. It was noted that among twenty described syndromes manifested with congenital heart disease, seven are currently referred to the group of primary immunodeficiencies. Types of congenital heart defects in the study group ranged from disorders of the valve apparatus to the non-functional opening between heart chambers. Signs of immune-related pathology included decreased thymus size revealed by ultrasound examination as well as lowered value of genetic markers TREC and KREC, retrospectively obtained from neonatal dry blood spots. Finally, we name a key element for successful differential diagnostics of PID with congenital heart disease – awareness of various medical workers about such pathology for developing algorithms for their interaction with regard to management of such comorbid patients.

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Przeździecki, R., M. Czopowicz i R. Sapierzyński. "Cytomorphometry of serosal effusion in dogs". Polish Journal of Veterinary Sciences 18, nr 3 (1.09.2015): 481–87. http://dx.doi.org/10.1515/pjvs-2015-0063.

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AbstractCytomorphometry made on cytological slides is the quantitative method of precise analysis of cellular structures, including both cytoplasm and nucleus. The aim of this study was to describe cytomorphometric parameters of mesothelial cells in the course of benign reactive and malignant proliferation and to compare them to carcinomas and adenocarcinomas located within serosal cavities in dogs. The second aim was to evaluate applicability of cytomorphometry to diagnostics of diseases causing accumulation of effusion in serosal cavities. Cytological samples of normal and non-malignant mesothelium, mesothelioma and various carcinomas were collected from dogs. Cytomorphometry was made on the smears stained with Giemsa solution. Mean nuclear and cellular perimeter, mean nuclear and cellular area, mean nuclear and cellular diameter, and mean nuclear and cellular roundness were determined. Moreover, nuclear to cytoplasmic ratio (N/C) was calculated. The data revealed statistically significant differences for all parameters, excluding mean nuclear perimeter, between compared groups. Normal mesothelium cells and their nuclei were significantly smaller and more elongated than cells and nuclei of both benign reactive and malignant neoplastic mesothelium. Only a few differences were observed between benign reactive mesothelium cells and mesothelioma cells – mean nuclear area and mean nuclear diameter of benign reactive mesothelium cells were significantly larger and N/C ratio was higher in comparison to mesothelioma cells. Even though some significant differences were observed, considerable overlap of these cytomorphometric parameters in animals with different diseases limited practical role of these observations. Cytomorphometric analysis of cellular samples collected from dogs with proliferative processes affecting serosal cavities can be only an auxiliary method increasing accuracy of preoperative diagnosis.

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Arinawati, Dian Yosi, i Aprilinda Widyawati. "Saliva sebagai Media Diagnosis untuk Deteksi Keganasan". STOMATOGNATIC - Jurnal Kedokteran Gigi 19, nr 2 (31.10.2022): 77. http://dx.doi.org/10.19184/stoma.v19i2.34728.

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Human saliva is a potential diagnostic fluid where most of changes in the body can be reflected in saliva and it considered a "mirror of the body". Currently, salivary biomarkers help in the detection of oral cancer, dental caries, periodontal disease, diabetes, breast cancer, and lung cancer. Saliva like others body liquid in human body such as urine and blood contain tumour derived biomarkers i.e DNA, asam amino, cells and vesicles which shed via bloodstream and secreted through salivary gland. Salivary diagnostic is a growing dynamic field utilizing nanotechnology and molecular diagnostics to aid in the diagnosis of oral and systemic diseases. The purpose of this review is to determine saliva as a diagnostic medium through various its biomarkers to detect malignancies in the body. The use of saliva as a medium for molecular diagnosis has the advantage of being sensitive, specific, can be used as a screening medium in diagnosing and staging, therefore usefull for early detection of malignancy, for example Oral Squamous Cell Carcinoma can be seen in three levels, changes in cellular DNA, altered mRNA transcripts, altered protein levels (intracellular or extracellular). It can also detect other cancers such as Salivary Gland Cancer, and even in parts of the body far from the oral cavity such as Lung Cancer, Pancreatic Cancer, Breast Cancer. It can be concluded that saliva can be used as a non-invasive diagnostic medium to detect various oral and systemic malignancies.

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Smolková, Barbora, Mariia Uzhytchak, Anna Lynnyk, Šárka Kubinová, Alexandr Dejneka i Oleg Lunov. "A Critical Review on Selected External Physical Cues and Modulation of Cell Behavior: Magnetic Nanoparticles, Non-thermal Plasma and Lasers". Journal of Functional Biomaterials 10, nr 1 (24.12.2018): 2. http://dx.doi.org/10.3390/jfb10010002.

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Physics-based biomedical approaches have proved their importance for the advancement of medical sciences and especially in medical diagnostics and treatments. Thus, the expectations regarding development of novel promising physics-based technologies and tools are very high. This review describes the latest research advances in biomedical applications of external physical cues. We overview three distinct topics: using high-gradient magnetic fields in nanoparticle-mediated cell responses; non-thermal plasma as a novel bactericidal agent; highlights in understanding of cellular mechanisms of laser irradiation. Furthermore, we summarize the progress, challenges and opportunities in those directions. We also discuss some of the fundamental physical principles involved in the application of each cue. Considerable technological success has been achieved in those fields. However, for the successful clinical translation we have to understand the limitations of technologies. Importantly, we identify the misconceptions pervasive in the discussed fields.

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Walker, Jillian Marie, Padraic O’Malley i Mei He. "Applications of Exosomes in Diagnosing Muscle Invasive Bladder Cancer". Pharmaceutics 14, nr 10 (23.09.2022): 2027. http://dx.doi.org/10.3390/pharmaceutics14102027.

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Muscle Invasive Bladder Cancer (MIBC) is a subset of bladder cancer with a significant risk for metastases and death. It accounts for nearly 25% of bladder cancer diagnoses. A diagnostic work-up for MIBC is inclusive of urologic evaluation, radiographic imaging with a CT scan, urinalysis, and cystoscopy. These evaluations, especially cystoscopy, are invasive and carry the risk of secondary health concerns. Non-invasive diagnostics such as urine cytology are an attractive alternative currently being investigated to mitigate the requirement for cystoscopy. A pitfall in urine cytology is the lack of available options with high reliability, specificity, and sensitivity to malignant bladder cells. Exosomes are a novel biomarker source which could resolve some of the concerns with urine cytology, due to the high specificity as the surrogates of tumor cells. This review serves to define muscle invasive bladder cancer, current urine cytology methods, the role of exosomes in MIBC, and exosomes application as a diagnostic tool in MIBC. Urinary exosomes as the specific populations of extracellular vesicles could provide additional biomarkers with specificity and sensitivity to bladder malignancies, which are a consistent source of cellular information to direct clinicians for developing treatment strategies. Given its strong presence and differentiation ability between normal and cancerous cells, exosome-based urine cytology is highly promising in providing a perspective of a patient’s bladder cancer.

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Raju Paul, Susan, Alexander Bagaev, Ivan Valiev, Vladimir Zyrin, Aleksandr Zaitsev, Daniyar Dyykanov, Katerina Nuzhdina i in. "Non-small cell lung cancer: Analysis using mass cytometry and next generation sequencing reveals new opportunities for the development of personalized therapies." Journal of Clinical Oncology 38, nr 15_suppl (20.05.2020): e21026-e21026. http://dx.doi.org/10.1200/jco.2020.38.15_suppl.e21026.

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e21026 Background: Comprehensive molecular profiling and the use of biomarkers as companion diagnostics have transformed precision medicine for cancer patients. To identify patient-specific tumor microenvironment and biomarker profiles, we assessed the accuracy of our deconvolution algorithm in identifying cellular compositions from whole exome (WES) and whole transcriptome (RNA-seq) sequencing of solid tumors compared with cell populations identified by Mass Cytometry by Time of Flight (CyTOF) in surgically resected tissue from non-small cell lung cancer (NSCLC) patients. Methods: Resected NSCLC tissue was divided for RNA-seq and WES of whole tissue (n = 9) and for generating tissue single cell suspensions through mechanical dissociation and enzymatic digestion (n = 11). Bulk RNA-seq and CyTOF were performed on all cell suspensions. Cellular phenotypes were identified using clustering algorithms in CyTOF and predicted from bulk RNA-seq using our proprietary computational method. Results: Cellular composition reconstructed from RNA-seq correlated with the composition detected by CyTOF (R2= 0.922, n = 7) from cell suspensions. To recover the cell percentage from bulk RNA-seq, a machine learning framework was trained on the cell compendia comprising 7,117 unique cell type RNA-seq profiles. A two-stage hierarchical learning procedure generated a gradient boosting Light GBM model that included training on artificial RNA-seq mixtures of different cell types. With this method, we found that stromal and malignant cells were depleted during single cell suspension preparation, resulting in statistically significant differences in the tumor cell composition reconstructed from solid tissue and single cell suspensions. Immune cell types namely T cells and macrophages were similarly represented in both the bulk tumor tissue and matched single cell suspensions. Transcriptomics revealed a subgroup of patients whose tumors were B-cell-enriched, which was validated in other NSCLC cohorts and was associated with greater CD4+ and CD8+ T cell infiltration and improved clinical outcomes. Conclusions: Since preparation of single cell suspensions leads to the loss of several cellular components, RNA-seq of tumor bulk tissue better describes the molecular and cellular properties of the tumor microenvironment. The combination of RNA-seq and WES of tumor tissue provides a comprehensive profile of cellular composition, suggesting that this combination is ideal for precision medicine applications.

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Szabó, Edit, Anna Kulin, Bálint Jezsó, Nóra Kucsma, Balázs Sarkadi i György Várady. "Selective Fluorescent Probes for High-Throughput Functional Diagnostics of the Human Multidrug Transporter P-Glycoprotein (ABCB1)". International Journal of Molecular Sciences 23, nr 18 (13.09.2022): 10599. http://dx.doi.org/10.3390/ijms231810599.

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The multidrug transporter ABCB1 (MDR1, Pgp) plays an important role in the absorption, distribution, metabolism, and elimination of a wide range of pharmaceutical compounds. Functional investigation of the ABCB1 expression is also essential in many diseases, including drug-resistant cancer, inflammatory conditions, or Alzheimer disease. In this study, we examined the potential interaction of the ABCB1 multidrug transporter with a group of commercially available viability dyes that are generally considered not to penetrate into intact cells. Here, we demonstrate that the slow cellular accumulation of TO-PRO™-1 (TP1) or TO-PRO™-3 (TP3) was strongly inhibited by ABCB1-dependent dye extrusion. TP1/3 dye accumulation was not affected by the presence of ABCC1 or ABCG2, while this uptake was increased to the level in the ABCB1-negative cells by a specific P-glycoprotein inhibitor, Tariquidar. We suggest that TP compounds can be used as highly sensitive, selective, non-toxic, and stable dyes to examine the functional expression and properties of the ABCB1 multidrug transporter, especially in microplate-based high-throughput flow cytometry assays. In addition, we demonstrate the applicability of the TP dyes to efficiently select and separate even a very low number of Pgp-expressing intact cells.

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Upadhyay, Ravi Kant, i Priya Rai. "Radiation induced therapeutic effects in cancerous and tumor cells: A review". Journal of Stem Cell Research & Therapeutics 8, nr 1 (2.02.2023): 1–12. http://dx.doi.org/10.15406/jsrt.2023.08.00156.

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Present review article describes use of radiation and radionuclides on cancer and cancer cell therapeutics. It also sketches out cumulative effects of radiation exposure received by the patients during cancer diagnostics. Though, in cancer therapeutics a selected and permissible dose is provided in several cycles to ablate the neoplastic cells and improve the condition of patient, but radiation harms surrounding cells and imparts negative effects on biology of cells. Ionizing radiation (IR) promotes cancer cell death through cytotoxicity. This article emphasizes both remedial effects and biological effects of radiation and radio-resistance in cells. It suggests safe use of radionucleides by encapsulating them in nanomaterials so as to use it alternate to chemotherapy to destroy various cancer types to enhance the survival of normal cells. This article explains effect of ionizing and non-ionizing radiation on cellular metabolism and genetics.

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Nassar, Samuel F., Khadir Raddassi, Baljit Ubhi, Joseph Doktorski i Ahmad Abulaban. "Precision Medicine: Steps along the Road to Combat Human Cancer". Cells 9, nr 9 (9.09.2020): 2056. http://dx.doi.org/10.3390/cells9092056.

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The diagnosis and treatment of diseases such as cancer is becoming more accurate and specialized with the advent of precision medicine techniques, research and treatments. Reaching down to the cellular and even sub-cellular level, diagnostic tests can pinpoint specific, individual information from each patient, and guide providers to a more accurate plan of treatment. With this advanced knowledge, researchers and providers can better gauge the effectiveness of drugs, radiation, and other therapies, which is bound to lead to a more accurate, if not more positive, prognosis. As precision medicine becomes more established, new techniques, equipment, materials and testing methods will be required. Herein, we will examine the recent innovations in assays, devices and software, along with next generation sequencing in genomics diagnostics which are in use or are being developed for personalized medicine. So as to avoid duplication and produce the fullest possible benefit, all involved must be strongly encouraged to collaborate, across national borders, public and private sectors, science, medicine and academia alike. In this paper we will offer recommendations for tools, research and development, along with ideas for implementation. We plan to begin with discussion of the lessons learned to date, and the current research on pharmacogenomics. Given the steady stream of advances in imaging mass spectrometry and nanoLC-MS/MS, and use of genomic, proteomic and metabolomics biomarkers to distinguish healthy tissue from diseased cells, there is great potential to utilize pharmacogenomics to tailor a drug or drugs to a particular cohort of patients. Such efforts very well may bring increased hope for small groups of non-responders and those who have demonstrated adverse reactions to current treatments.

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Kothmaier, Hannelore, Daniela Rohrer, Elvira Stacher, Franz Quehenberger, Karl-Friedrich Becker i Helmut H. Popper. "Comparison of Formalin-Free Tissue Fixatives: A Proteomic Study Testing Their Application for Routine Pathology and Research". Archives of Pathology & Laboratory Medicine 135, nr 6 (1.06.2011): 744–52. http://dx.doi.org/10.5858/2009-0676-oa.1.

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Abstract Context.—Formalin-fixed, paraffin-embedded tissue is the routine processing method for diagnostics practiced in pathology departments worldwide. Objective.—To determine the potential value of non–cross-linking, formalin-free tissue fixation for diagnostics in pathology and proteomic investigations. Design.—We tested 3 commercially available, formalin-free tissue fixatives—FineFIX, RCL2, and HOPE—in lung cancer specimens from 10 patients. The fixatives were evaluated for their effects on tissue morphology, protein recovery, and immunoreactivity for a selected panel of proteins differently expressed in lung cancer, using immunohistochemistry and Western blotting. Results.—Tumor-cell analysis with hematoxylin-eosin worked equally well for all tested fixatives when compared with the standard formalin-fixed, paraffin-embedded procedure. Movat pentachrome stains showed comparable results for the different matrices and cellular proteins analyzed. The RCL2 (P = .01) and HOPE fixatives (P = .03) improved protein recovery when compared with formalin-fixed, paraffin-embedded or frozen tissues. Our data clearly show that the fixatives evaluated influenced immunoreactivity to matched, formalin-fixed, paraffin-embedded lung cancer tissue. In particular, membrane-bound proteins, such as epidermal growth factor receptor EGFR, can be detected more efficiently by immunohistochemistry and Western blotting. Conclusion.—We have demonstrated that formalin-free fixatives have the potential in routine pathology and research to replace formalin in histomorphology and protein preservation.

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Evangelopoulos, Michael, Alessandro Parodi, Jonathan Martinez i Ennio Tasciotti. "Trends towards Biomimicry in Theranostics". Nanomaterials 8, nr 9 (21.08.2018): 637. http://dx.doi.org/10.3390/nano8090637.

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Over the years, imaging and therapeutic modalities have seen considerable progress as a result of advances in nanotechnology. Theranostics, or the marrying of diagnostics and therapy, has increasingly been employing nano-based approaches to treat cancer. While first-generation nanoparticles offered considerable promise in the imaging and treatment of cancer, toxicity and non-specific distribution hindered their true potential. More recently, multistage nanovectors have been strategically designed to shield and carry a payload to its intended site. However, detection by the immune system and sequestration by filtration organs (i.e., liver and spleen) remains a major obstacle. In an effort to circumvent these biological barriers, recent trends have taken inspiration from biology. These bioinspired approaches often involve the use of biologically-derived cellular components in the design and fabrication of biomimetic nanoparticles. In this review, we provide insight into early nanoparticles and how they have steadily evolved to include bioinspired approaches to increase their theranostic potential.

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Singh, Desh Deepak, Youngsun Kim, Seung Ah Choi, Ihn Han i Dharmendra Kumar Yadav. "Clinical Significance of MicroRNAs, Long Non-Coding RNAs, and CircRNAs in Cardiovascular Diseases". Cells 12, nr 12 (14.06.2023): 1629. http://dx.doi.org/10.3390/cells12121629.

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Based on recent research, the non-coding genome is essential for controlling genes and genetic programming during development, as well as for health and cardiovascular diseases (CVDs). The microRNAs (miRNAs), lncRNAs (long ncRNAs), and circRNAs (circular RNAs) with significant regulatory and structural roles make up approximately 99% of the human genome, which does not contain proteins. Non-coding RNAs (ncRNA) have been discovered to be essential novel regulators of cardiovascular risk factors and cellular processes, making them significant prospects for advanced diagnostics and prognosis evaluation. Cases of CVDs are rising due to limitations in the current therapeutic approach; most of the treatment options are based on the coding transcripts that encode proteins. Recently, various investigations have shown the role of nc-RNA in the early diagnosis and treatment of CVDs. Furthermore, the development of novel diagnoses and treatments based on miRNAs, lncRNAs, and circRNAs could be more helpful in the clinical management of patients with CVDs. CVDs are classified into various types of heart diseases, including cardiac hypertrophy (CH), heart failure (HF), rheumatic heart disease (RHD), acute coronary syndrome (ACS), myocardial infarction (MI), atherosclerosis (AS), myocardial fibrosis (MF), arrhythmia (ARR), and pulmonary arterial hypertension (PAH). Here, we discuss the biological and clinical importance of miRNAs, lncRNAs, and circRNAs and their expression profiles and manipulation of non-coding transcripts in CVDs, which will deliver an in-depth knowledge of the role of ncRNAs in CVDs for progressing new clinical diagnosis and treatment.

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Saraswathi S, Sushil Kumar Middha i Jolitha B. "An Integrated Approach to Diagnosis and Therapy in Cancer". International Journal of Fundamental and Applied Sciences (IJFAS) 6, nr 4 (30.12.2017): 1–30. http://dx.doi.org/10.59415/ijfas.v6i4.111.

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Cancer is one of the major non-communicable diseases (NCD) worldwide, accounting for 22% deaths in 2012 worldwide. The number of cases is projected to increase to 17 million by 2020. According to ICMR, almost half of the cases will be in Asia with more than 17 lakh cases expected to occur in India by 2020. In such a scenario, early diagnosis and cost-effective treatment will play a major role in effective patient management. Developing the infrastructure required to cater to a vast population requires substantial effort in research and development. The National Symposium on “An Integrated Approach to Diagnosis and Therapy in Cancer” aims to create awareness about cancer etiology and stimulate the interest of students in cancer research. It has become unequivocally evident that tumor development depends on the intricate reciprocal interplay of tumor cells with their local and distant environments. Mutations in genes regulating cell cycle/cell proliferation can also contribute to tumorigenesis and is of great relevance in both diagnosis and disease management. One of the major focus of the symposia is developing diagnostics and therapeutics through translational research. The invited speakers will make presentations on emerging areas in cancer research like nanomedicine, personalized medicine, etc. Reputed clinicians and researchers will provide different perspectives on cancer biology. Thus, this symposium aims at a comprehensive functional understanding on the integrated role of physical and life sciences in diagnostics and therapeutics of cellular and molecular events that are responsible for the plasticity of tumor cells. Moreover, it will bring together experts in cancer and molecular biology who will share their expertise on a new generation of effective diagnostic tools and therapeutic approaches in cancer. One of the objectives of the symposium is to provide students and researchers with a platform for the exchange of knowledge and innovative ideas towards a comprehensive approach in cancer biology. It provides them with an ample opportunity to present their research work related to the conference theme.

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Kostyuchenko, M. V., E. L. Rakitina, D. G. Ponomarenko, O. V. Logvinenko, S. A. Kurcheva, T. V. Berdnikova, D. V. Rusanova i A. N. Kulichenko. "STUDYING DEVELOPMENT OF POST-VACCINAL CELLULAR IMMUNITY AGAINST BRUCELLOSIS BY MEANS OF LYMPHOCYTE IN VITRO TESTS USING AN EXPERIMENTAL ANTIGENIC COMPLEX". Medical Immunology (Russia) 21, nr 3 (13.07.2019): 547–54. http://dx.doi.org/10.15789/1563-0625-2019-3-547-554.

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Regulatory framework and methodological approaches to evaluation of immunological effects of vaccination against brucellosis are not established, and the degree of immunological post-vaccinal rearrangement is not yet developed. Due to leading role of cellular immunity in formation of immune protection against brucellosis, evaluation the cellular response in response to antigenic stimulation may be considered the most informative and objective approach to analysis of immune changes in the body during vaccination. In order to develop the most diagnostically informative methods for design of antigen-stimulation cell tests in vitro, a careful selection of a stimulating agent (antigen) is required, which should have a sufficient activating potential, thus providing specificity of reaction under in vitro conditions. The aim of the present study is to study the in vitro specific activity of a protein-polysaccharide antigenic complex from the Brucella abortus 19 BA strain (BrAg), and an opportunity of its application in order to assess the formation of post-vaccinal cellular immunity against brucellosis.The study was performed with white laboratory mice (n = 50) immunized with the Brucella abortus 19 BA strain. The control group (n = 50) consisted of laboratory mice that received a sterile saline solution in a volume of 0.5 ml. Blood samples were taken from immunized and control animals before vaccination, and 7, 14, 21, and 30 days after immunization. By means of flow cytometry, the activation molecules CD25, CD69, MHC II and CD95, expressed on T lymphocytes (CD3+CD69+, CD3+CD25+, CD3+CD95+, CD3+MHC+) were determined. To observe the development of immunity, the intensity of expression of T lymphocyte activation markers was calculated using the stimulation quotient. BrAg was used for specific in vitro stimulation of T lymphocytes. The liquid brucellosis allergen (brucellin) was used as an antigen for comparison, when studying opportunity of BrAg usage for assessing the postvaccinal immunity development.The following results were obtained: BrAg has pronounced specific activity, it did not cause non-specific in vitro reactions (activation) of T lymphocytes, thus enabling its application as a test antigen when evaluating development of adaptive vaccine immunity against brucella.Experimental testing of brucellosis antigen for carrying out the in vitro antigen-stimulated cellular reactions, aiming for evaluation of post-vaccinal immunity development against brucellosis, showed that the usage of BrAg promotes increase in diagnostic sensitivity of cellular reactions under in vitro experimental conditions. The applied experimental antigen is a quite promising tool for development of laboratory algorithms for brucellosis diagnostics, and assessment of actual vaccination efficiency in cohorts previously vaccinated against brucellosis.

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Dolhikh, H. V., H. S. Maslak, V. I. Didenko, I. A. Klenina i А. О. Dolhikh. "Levels of isoforms of fibronectin and α5/CD49e integrin on lymphocytes and in blood plasma in the conditions of chronic diffuse liver diseases". Regulatory Mechanisms in Biosystems 11, nr 4 (27.10.2020): 475–82. http://dx.doi.org/10.15421/022073.

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Chronic diffuse liver diseases are characterized by accumulation of complex inflammatory infiltrate in the liver tissues, blood, and lympha, and activation of the immune system. Leukocytes become involved in the area of inflammation after the activation of receptors of blood adhesia, particularly integrins and their ligands. Plasma lymphocytes quickly activate the function of integrins by changing their conformation, leading to high affinity and underlying the formation of strong stable connection between the components of extracellular matrix. A vitally important role in the process of liver fibrogenesis is performed by a pro-fibrogenicic protein fibronectin which induces the expresson of collagen genes and precedes the deposition of other components of matrix. The studies were conducted in the group of patients suffering from chronic diffuse liver diseases of non-viral etiology aged 28–60 years, n = 36 and in the group of 15 practically healthy volunteer donors aged 25 to 52 years without a history of liver diseases using the methods of flow cytofluorometry, immunoenzymatic analysis, and quantitative real-time polymerase chain reaction. The patients of the group with chronic diffuse liver diseases were observed to have statistically significant decrease in the concentration of plasmatic form of fibronectin measuring 27.6% compared with the control group. We determined increase in the concentration of cellular fibronectin in blood plasma of patients with the diseases on average accounting for 63.8% compared with the norm, and the highest increase in this parameter equaling 77.2% was seen in patients suffering from drug-induced hepatitis. Significant increase in the level of exposure of cellular FN on blood lymphocytes was determined in patients with chronic diffuse liver diseases, measuring 231.8%, whereas the level of plasmatic form of fibronectin in these cells was decreased (statistically unreliable). For α5-integrin subunit, we determined a 390.8% increase in the level of its exposure in blood lymphocytes in the surveyed groups compared with the control. Level of blood lymphocytes that express the cellular fibronectin significantly decreased by 140.1%. Statistical characteristics of diagnostic possibility of the parameters of the level of plasmatic and cellular fibronectin in blood, determined over the analysis of ROC-curves, demonstrated excellent informativeness of these tests. Analysis of the possibility of predicting the presence of pathology using the model of logistic regression revealed zero error of prediction and maximum efficiency of the tests: intensity of exposure of α5-integrin receptor on the surface of lymphocytes, intensity of exposure of plasmatic fibronectin on the surface of lymphocytes, intensity of exposure of cellular fibronectin on the surface of lymphocytes, concentration of plasmatic fibronectin in blood, concentration of cellular fibronectin in blood plasma. These parameters may be proposed for further surveys for developing serologic biomarkers based on the parameters for diagnostics of chronic diffuse liver diseases.

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Surguchov, Andrei. "Invertebrate Models Untangle the Mechanism of Neurodegeneration in Parkinson’s Disease". Cells 10, nr 2 (16.02.2021): 407. http://dx.doi.org/10.3390/cells10020407.

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Parkinson’s disease (PD) is the second most common neurodegenerative disease, afflicting ~10 million people worldwide. Although several genes linked to PD are currently identified, PD remains primarily an idiopathic disorder. Neuronal protein α-synuclein is a major player in disease progression of both genetic and idiopathic forms of PD. However, it cannot alone explain underlying pathological processes. Recent studies demonstrate that many other risk factors can accelerate or further worsen brain dysfunction in PD patients. Several PD models, including non-mammalian eukaryotic organisms, have been developed to identify and characterize these factors. This review discusses recent findings in three PD model organisms, i.e., yeast, Drosophila, and Caenorhabditis elegans, that opened new mechanisms and identified novel contributors to this disorder. These non-mammalian models share many conserved molecular pathways and cellular processes with humans. New players affecting PD pathogenesis include previously unknown genes/proteins, novel signaling pathways, and low molecular weight substances. These findings might respond to the urgent need to discover novel drug targets for PD treatment and new biomarkers for early diagnostics of this disease. Since the study of neurodegeneration using simple eukaryotic organisms brought a huge amount of information, we include only the most recent or the most important relevant data.

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Jandhyala, Sidhartha, Austin Van Namen, Catalina-Paula Spatarelu i Geoffrey P. Luke. "EGFR-Targeted Perfluorohexane Nanodroplets for Molecular Ultrasound Imaging". Nanomaterials 12, nr 13 (30.06.2022): 2251. http://dx.doi.org/10.3390/nano12132251.

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Perfluorocarbon nanodroplets offer an alternative to gaseous microbubbles as contrast agents for ultrasound imaging. They can be acoustically activated to induce a liquid-to-gas phase transition and provide contrast in ultrasound images. In this study, we demonstrate a new strategy to synthesize antibody-conjugated perfluorohexane nanodroplet (PFHnD-Ab) ultrasound contrast agents that target cells overexpressing the epidermal growth factor receptor (EGFR). The perfluorohexane nanodroplets (PFHnD) containing a lipophilic DiD fluorescent dye were synthesized using a phospholipid shell. Antibodies were conjugated to the surface through a hydrazide-aldehyde reaction. Cellular binding was confirmed using fluorescence microscopy; the DiD fluorescence signal of the PFHnD-Ab was 5.63× and 6× greater than the fluorescence signal in the case of non-targeted PFHnDs and the EGFR blocking control, respectively. Cells were imaged in tissue-mimicking phantoms using a custom ultrasound imaging setup consisting of a high-intensity focused ultrasound transducer and linear array imaging transducer. Cells with conjugated PFHnD-Abs exhibited a significantly higher (p < 0.001) increase in ultrasound amplitude compared to cells with non-targeted PFHnDs and cells exposed to free antibody before the addition of PFHnD-Abs. The developed nanodroplets show potential to augment the use of ultrasound in molecular imaging cancer diagnostics.

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Fedulova, Liliya V., Alexandr A. Basov, Ekaterina R. Vasilevskaya i Stepan S. Dzhimak. "Gender Difference Response of Male and Female Immunodeficiency Rats Treated with Tissue-specific Biomolecules". Current Pharmaceutical Biotechnology 20, nr 3 (30.04.2019): 245–53. http://dx.doi.org/10.2174/1389201020666190222184814.

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Background:The modern immunology is targeted to the detailed study of various immunopathological conditions at the molecular and cellular level, development of new methods for the prevention, diagnostics and treatment of contagious and non-contagious diseases of humans and animals.Methods:In the present work we took the rats with model of cyclophosphamide-induced immunodeficiency and studied the features of gender impact of the complex extract of immunocompetent organs (thymus, spleen and mesenteric lymph nodes) Sus scrofa and its separate fraction with molecular weight less than 30 kDa administered to male and female rats.Results:The impact of gender differences and tissue-specific biomolecules (30 kDa fraction) on hematological parameters (leukocytes, erythrocytes, platelets), functional activity of immune system (IL-2, IL-4, IL-6, complement system, IgG, IgM), biochemical parameters of hepatocytes functioning (activity of ALP and LDG), carbohydrate metabolism (glucose) and lipid metabolism (triglycerides).Conclusion:Decrease of ALP activity is caused by inhibition of bile formation in a liver after introduction of cytostatic agent, and in contrast to complex extract, the administration of fraction 30 kDa allows improving bile production in male rats.

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Deshwal, Himanshu, Tatiana Weinstein i Roxana Sulica. "Advances in the management of pulmonary arterial hypertension". Journal of Investigative Medicine 69, nr 7 (27.09.2021): 1270–80. http://dx.doi.org/10.1136/jim-2021-002027.

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The management of pulmonary arterial hypertension (PAH) has significantly evolved over the last decades in the wake of more sensitive diagnostics and specialized clinical programs that can provide focused medical care. In the current era of PAH care, 1-year survival rates have increased to 86%–90% from 65% in the 1980s, and average long-term survival has increased to 6 years from 2.8 years. The heterogeneity in the etiology and disease course has opened doors to focusing research in phenotyping the disease and understanding the pathophysiology at a cellular and genetic level. This may eventually lead to precision medicine and the development of medications that may prevent or reverse pulmonary vascular remodeling. With more insight, clinical trial designs and primary end-points may change to identify the true survival benefit of pharmacotherapy. Identifying responders from non-responders to therapy may help provide individualized patient-centered care rather than an algorithm-based approach. The purpose of this review is to highlight the latest advances in screening, diagnosis, and management of PAH.

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Dyakova, M. E., K. B. Vladimirov, D. S. Esmedlyaeva i P. K. Yablonskiy. "Enzymes of purine metabolism — biomarkers for the diagnostics of tuberculous pleurisy in patients with HIV infection". HIV Infection and Immunosuppressive Disorders 15, nr 1 (30.04.2023): 32–40. http://dx.doi.org/10.22328/2077-9828-2023-15-1-32-40.

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The objective of the study was to evaluate the information content of determining the activity of adenosine deaminase and adenosine deaminase-2 in the diagnosis of tuberculous pleurisy in patients with HIV infection.Materials and methods. A total of 378 patients with pleural effusion were retrospectively examined. In 215 cases, tuberculous pleurisy was detected (TP); and 163 patients had non-tuberculous pleural effusion (non-TP). As much as 27 patients in the TP group were HIV co-infected (TP/HIV+), the remaining 188 patients were HIV — negative (TP/HIV–). In all the patients, the activity of total adenosine deaminase (ADA) and its isoenzymes (ADA-1 and ADA-2) in the pleural fluid was determined.Results and discussion. In the TP group, the activity of total ADA (95.5 [67.7; 115.4] versus 82.0 [59.6; 100.0] U/L, p=0.1), ADA-1 (14.2 [5.8; 20.5] versus 12.1 [6.1; 23.7] U/L, p=0.9) and ADA-2 (78,1 [38.1; 93.1] versus 62.4 [35.4; 82.2] U/L, p=0,1) did not depend on HIV status. The activity of these indicators was determined above the threshold level — total ADA in 96.3% and 95.2%, ADA-1 in 25.9% and 30.8% and ADA-2 in 92.6% and 83.3% of cases in the «TP/HIV+» and «TP/HIV–» groups, respectively. A negative correlation between ADA-1 activity and HIV viral load in the group of patients with tuberculous pleurisy and HIV infection (r=–0.45; p=0.008), as well as in the subgroup of TP/HIV+ patients who received (r=–0.9; p=0.008) and in those who didn’t receive ART (r=–0.47; p=0.04) was obtained. Our results show that a total ADA activity increase in the patients with tuberculous pleurisy, regardless of patients’ HIV status, occur due to ADA-2. Thus, the increase in activity of total ADA and ADA-2 in our study was caused by active tuberculosis, not by the presence or absence of HIV co-infection. Also, the ADA-2 activity in HIV-infected patients is likely consistent with ADA-2 important role in cellular immune responses.Conclusion. Our data indicate the participation of purine metabolism enzymes in the pathogenesis of HIV infection. At the same time, adenosine deaminase activity is not a specific biomarker of individual changes characteristic of HIV infection. The study results suggest that the total adenosine deaminase and adenosine deaminase-2 activity increase is a valuable and diagnostically significant marker of tuberculous pleurisy in HIV-infected patients. The value of adenosine deaminase and adenosine deaminase-2 activity remains high even in the patients having severe immunosuppression, which allows them to be actively used for rapid diagnostics and hence, early TB therapy initiation.

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Straus, Don, Ann Zuniga, Alejandra Garces, Andrew Tempesta, Adam Williams, Bill Lauzier, Jennifer Hickey i in. "Rapid Ultrasensitive Detection of Clostridiodes difficile Toxins in Stool Samples Using A Single-Molecule Counting Method". Infection Control & Hospital Epidemiology 41, S1 (październik 2020): s450—s451. http://dx.doi.org/10.1017/ice.2020.1121.

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Background:Clostridiodes difficile infection is considered an urgent antibiotic resistance threat by the CDC, accounting for ∼225,000 hospitalizations, 12,800 deaths, and ∼$1 billion in healthcare costs in the United States in 2017. The presence of the secreted toxins that cause the devastating symptoms of this gastrointestinal infection are diagnostic of C. difficile infection (CDI). However, the rapid testing methods currently used to detect CDI lack accuracy. Enzyme immunoassays are specific but lack sensitivity because they do not detect CDI patients that have low levels of the toxins. Nucleic acid amplification tests (NAATs) are sensitive, but they lack specificity because they detect patients colonized with C. difficile in the dormant spore form that does not produce the toxins. This insufficiency has resulted in the adoption of complex multitest algorithms for C. difficile diagnosis. We present results for a new toxin test that demonstrates both high clinical sensitivity and clinical specificity for C. difficile toxin B on a fully automated benchtop platform. Methods: The detection technology uses nonmagnified digital imaging to count single toxin molecules that tether together target-specific magnetic and fluorescent particles. The 30-minute method includes the use of a dye cushion to eliminate wash steps and the need for time-consuming specimen preparation steps. We determined analytical performance characteristics of the test using negative clinical stool samples spiked with purified toxin. To assess clinical performance, we tested 785 stool samples from 5 clinical sites and compared the results with the cellular cytotoxicity neutralization assay (CCNA). Results: The test’s limit of detection for toxin B was 60 pg/mL. A comparison of the new test to the CCNA reference method gave 98% positive percentage agreement (83 of 85 samples) and 95% negative percentage agreement (667 of 700 samples). Conclusions: The new method demonstrated 96% accuracy compared to the gold standard for C. difficile toxin tests. The results also demonstrate an analytical sensitivity (limit of detection, 60 pg/mL). Thus, the test has the potential to detect CDI patients missed by enzyme immunoassay (EIA) tests due to their low analytical sensitivity. Because the test detects toxins directly, it is expected to have a lower false-positive rate than NAAT methods, which detect patients colonized with the non–toxin-producing spore form. A single accurate test for toxin-producing C. difficile could eliminate the need for multitest algorithms.Funding: First Light Diagnostics, Inc., provided support for this study.Disclosures: Donald Straus reports that he is the founder and chief scientific officer of First Light Diagnostics (FLDx) with salary and ownership interest in the form of stocks, stock options, and warrants. Adam Williams reports salary from First Light Diagnostics.

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Guglas, Kacper, Tomasz Kolenda, Anna Teresiak, Magda Kopczyńska, Izabela Łasińska, Jacek Mackiewicz, Andrzej Mackiewicz i Katarzyna Lamperska. "lncRNA Expression after Irradiation and Chemoexposure of HNSCC Cell Lines". Non-Coding RNA 4, nr 4 (14.11.2018): 33. http://dx.doi.org/10.3390/ncrna4040033.

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Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cause of cancer mortality in the world. To improve the quality of diagnostics and patients’ treatment, new and effective biomarkers are needed. Recent studies have shown that the expression level of different types of long non-coding RNAs (lncRNAs) is dysregulated in HNSCC and correlates with many biological processes. In this study, the response of lncRNAs in HNSCC cell lines after exposure to irradiation and cytotoxic drugs was examined. The SCC-040, SCC-25, FaDu, and Cal27 cell lines were treated with different radiation doses as well as exposed to cisplatin and doxorubicin. The expression changes of lncRNAs after exposure to these agents were checked by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Target prediction was performed using available online tools and classified into specific biological processes and cellular pathways. The results indicated that the irradiation, as well as chemoexposure, causes changes in lncRNA expression and the effect depends on the cell line, type of agents as well as their dose. After irradiation using the dose of 5 Gy significant dysregulation of 4 lncRNAs, 10 Gy-5 lncRNAs, and 20 Gy-3 lncRNAs, respectively, were observed in all cell lines. Only lncRNAs Zfhx2as was down-regulated in all cell lines independently of the dose used. After cisplatin exposure, 14 lncRNAs showed lower and only two higher expressions. Doxorubicin resulted in lower expressions of eight and increased four of lncRNAs. Common effects of cytotoxic drugs were observed in the case of antiPEG11, BACE1AS, PCGEM1, and ST7OT. Analysis of the predicted targets for dysregulated lncRNAs indicated that they are involved in important biological processes, regulating cellular pathways connected with direct response to irradiation or chemoexposure, cellular phenotype, cancer initiating cells, and angiogenesis. Both irradiation and chemoexposure caused specific changes in lncRNAs expression. However, the common effect is potentially important for cellular response to the stress and survival. Further study will show if lncRNAs are useful tools in patients’ treatment monitoring.

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Zhelavskyi, M. M., S. P. Kernychnyi i O. Ya Dmytriv. "Сell death and its significance in reproductive pathology". Ukrainian Journal of Veterinary and Agricultural Sciences 4, nr 2 (21.06.2021): 18–26. http://dx.doi.org/10.32718/ujvas4-2.04.

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Since the middle of the last century, scientists have been interested in the mechanisms of regulation of cell division, differentiation and aging of cells. The first objects of study were insects, helminths and other living organisms. From the very beginning, in the biology of cell development and regulation, scientists have attached leading importance to genetic factors. Later, more and more experience was gained on the influence of intracellular factors, metabolic changes and exogenous pathogens on the programmed cell death. Recent research on cell biology and pathology has focused on the study of apoptosis. The first described phenomenon of programmed cell death was apoptosis. Subsequent studies were aimed at the study programmed cell death. This review will provide an opportunity to consider the biological mechanisms of programmed cell death, differences and species characteristics. The author described the clinical aspects of apoptosis, necroptosis and pyroptosis and their importance in the formation of cellular homeostasis. In the present review article simple classification system, where the cell death entities are primarily categorized into programmed cell death. Multiple mechanisms and phenotypes compose programmed non-apoptotic cell death, including: autophagy, entosis, methuosis and paraptosis, mitoptosis and parthanatos, ferroptosis, pyroptosis NETosis and necroptosis. Changes of cellular regulation at development of pathologies at people and animals are considered. Cell biology includes a variety of mechanisms of programmed aging and death. Modern research is aimed at deepening the study multiple mechanisms and phenotypes compose programmed. Cells. will certainly be taken into account by the Nomenclature Committee on Cell Death. Cellular regulation is associated with a variety of physiological mechanisms of development, and is also important in processes such as inflammation, immune response, embryogenesis maintenance of tissue homeostasis. Study of factors of influence and mechanisms of regulation of aging of cells opens a curtain for development of the newest means of diagnostics of pathologies and development of pharmacological means for correction of cellular mechanisms at development of pathologies.

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Tambuzzi, Stefano, Guendalina Gentile, Michele Boracchi, Domenico Di Candia, Rachele Bianchi i Riccardo Zoja. "Postmortem diagnostics of assumed suicidal food anaphylaxis in prison: a unique case of anaphylactic death due to peach ingestion". Forensic Science, Medicine and Pathology 17, nr 3 (3.05.2021): 449–55. http://dx.doi.org/10.1007/s12024-021-00373-1.

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AbstractSuicidal ingestion of food which the victim is aware they are allergic to is an exceptional occurrence in the forensic field. To the best of our knowledge, no cases of suicidal food anaphylaxis have been reported to date. Therefore we present the first case described in the literature. A 30-year-old prisoner was found dead inside his cell with the remains of a peach remains next to his body, and a handwritten farewell note in his pocket. The autopsy revealed only non-specific findings, while laboratory investigations (serological, toxicological, histological, and immunohistochemical) played a pivotal role in determing the cause and manner of death. In particular, a high titer of both total and specific IgE antibodies was detected, as well as an increase of the tryptase level in cadaveric blood. Moreover, a massive concentration of salicylates was measured in the gastric contents. Microscopically, cellular residues characterized by a vegetal structure were observed in the gastric contents and elements suggestive of mast cells were detected in the glottis, lungs, and myocardium. The immunohistochemical investigation with anti-CD117 and anti-tryptase antibodies showed positivity for mast cells, some of which appeared degranulated. Such findings were entirely consistent with an acute systemic anaphylactic reaction triggered by allergy. Therefore, the prisoner’s death was attributed to self-induced food anaphylaxis caused by the ingestion of peaches. This conclusion was achieved based only on circumstantial data, anamnestic information, autopsy findings, and multiple laboratory results. This integrated approach should be used to pursue a post-mortem diagnosis of anaphylaxis.

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