scholarly journals Presence of SARS-CoV-2 Nucleoprotein in Cardiac Tissues of Donors with Negative COVID-19 Molecular Tests

Diagnostics ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 731
Author(s):  
Gianluca Lorenzo Perrucci ◽  
Elena Sommariva ◽  
Veronica Ricci ◽  
Paola Songia ◽  
Yuri D’Alessandra ◽  
...  
Keyword(s):  
Tissue Bank ◽  
Digital Pcr ◽  
Tissue Donation ◽  
Vein Wall ◽  
Cardiac Tissues ◽  
Reverse Transcription Pcr ◽  
Molecular Tests ◽  
Negative Results ◽  
Italian Regions ◽  
Tissue Specimens

The 2019 Coronavirus disease (COVID-19) outbreak had detrimental effects on essential medical services such as organ and tissue donation. Lombardy, one of the most active Italian regions in organ/tissue procurement, has been strongly affected by the COVID-19 pandemic. To date, data concerning the risk of SARS-CoV-2 transmission after tissue transplantation are controversial. Here, we aimed to evaluate the presence/absence of SARS-CoV-2 in different cardiac tissues eligible for transplantation obtained from Lombard donors. We used cardiovascular tissues from eight donors potentially suitable for pulmonary valve transplantation. All donor subjects involved in the study returned negative results for the SARS-CoV-2 RNA molecular tests (quantitative real-time reverse-transcription PCR, qRT-PCR, and chip-based digital PCR) in nasopharyngeal swabs (NPS) or bronchoalveolar lavage (BAL). None of the eight donors included in this study revealed the presence of the SARS-CoV-2 viral genome. However, evaluation of the protein content of pulmonary vein wall (PVW) tissue revealed variable levels of SARS-CoV-2 nucleoprotein signal in all donors. Our study demonstrated for the first time, to the best of our knowledge, that viral nucleoprotein but not viral RNA was present in the examined tissue bank specimens, suggesting the need for caution and in-depth investigations on implantable tissue specimens collected during the COVID-19 pandemic period.

scholarly journals Using Molecular Diagnostics to Develop Therapeutic Strategies for Carbapenem-Resistant Gram-Negative Infections

2021 ◽  
Vol 11 ◽  
Author(s):  
Fred C. Tenover
Keyword(s):  
Carbapenem Resistance ◽  
Multidrug Resistant ◽  
Therapeutic Strategies ◽  
Beta Lactam ◽  
Gram Negative ◽  
Molecular Tests ◽  
Negative Results ◽  
Carbapenem Resistant ◽  
Class B ◽  
Global Threat

Infections caused by multidrug-resistant Gram-negative organisms have become a global threat. Such infections can be very difficult to treat, especially when they are caused by carbapenemase-producing organisms (CPO). Since infections caused by CPO tend to have worse outcomes than non-CPO infections, it is important to identify the type of carbapenemase present in the isolate or at least the Ambler Class (i.e., A, B, or D), to optimize therapy. Many of the newer beta-lactam/beta-lactamase inhibitor combinations are not active against organisms carrying Class B metallo-enzymes, so differentiating organisms with Class A or D carbapenemases from those with Class B enzymes rapidly is critical. Using molecular tests to detect and differentiate carbapenem-resistance genes (CRG) in bacterial isolates provides fast and actionable results, but utilization of these tests globally appears to be low. Detecting CRG directly in positive blood culture bottles or in syndromic panels coupled with bacterial identification are helpful when results are positive, however, even negative results can provide guidance for anti-infective therapy for key organism-drug combinations when linked to local epidemiology. This perspective will focus on the reluctance of laboratories to use molecular tests as aids to developing therapeutic strategies for infections caused by carbapenem-resistant organisms and how to overcome that reluctance.

Myocarditis: aetiology and histopathological diagnosis

ESC CardioMed ◽  
2018 ◽  
pp. 1512-1515
Author(s):  
Sabine Pankuweit
Keyword(s):  
Parvovirus B19 ◽  
Specific Treatment ◽  
Influenza Viruses ◽  
Histopathological Diagnosis ◽  
Cardiac Tissues ◽  
Barr Virus ◽  
Reverse Transcription Pcr ◽  
Epstein Barr ◽  
Inflammatory Infiltrates ◽  
Infiltrating Lymphocytes

Although the aetiology of myocarditis in a given patient often remains unknown, a large variety of infectious agents, including viruses, bacteria, protozoa, and fungi, systemic and autoimmune diseases, drugs, and toxins can cause myocarditis. The diagnosis of myocarditis is complex and challenging. The diagnostic gold standard is endomyocardial biopsy in which myocarditis is defined using the histological Dallas criteria and immunohistochemistry. Current diagnostic criteria require more than 14 infiltrating lymphocytes/mm2 (up to 4 macrophages may be included with the presence of CD3-positive T lymphocytes ≥7 cells/mm2). Inflammatory infiltrates are subdivided into lymphocytic, eosinophilic, polymorphic, giant cell, and cardiac sarcoidosis as these diagnoses imply a specific treatment and prognosis. Molecular biological analyses of cardiac tissues should include real-time polymerase chain reaction (PCR) and reverse transcription PCR assays for the most common cardiotropic infectious viruses such as enteroviruses, adenoviruses, cytomegalovirus, influenza viruses, human herpes virus 6, Epstein–Barr virus, hepatitis C virus, and parvovirus B19.

scholarly journals Establishment and Validation of RNA-Based Predictive Models for Understanding Survival of Vibrio parahaemolyticus in Oysters Stored at Low Temperatures

2017 ◽  
Vol 83 (6) ◽  
Author(s):  
Chao Liao ◽  
Yong Zhao ◽  
Luxin Wang
Keyword(s):  
Real Time ◽  
Predictive Models ◽  
Vibrio Parahaemolyticus ◽  
False Negative ◽  
Bacterial Inactivation ◽  
Rt Pcr ◽  
Content Type ◽  
Low Temperature Storage ◽  
Reverse Transcription Pcr ◽  
Negative Results

ABSTRACT This study developed RNA-based predictive models describing the survival of Vibrio parahaemolyticus in Eastern oysters (Crassostrea virginica) during storage at 0, 4, and 10°C. Postharvested oysters were inoculated with a cocktail of five V. parahaemolyticus strains and were then stored at 0, 4, and 10°C for 21 or 11 days. A real-time reverse transcription-PCR (RT-PCR) assay targeting expression of the tlh gene was used to evaluate the number of surviving V. parahaemolyticus cells, which was then used to establish primary molecular models (MMs). Before construction of the MMs, consistent expression levels of the tlh gene at 0, 4, and 10°C were confirmed, and this gene was used to monitor the survival of the total V. parahaemolyticus cells. In addition, the tdh and trh genes were used for monitoring the survival of virulent V. parahaemolyticus. Traditional models (TMs) were built based on data collected using a plate counting method. From the MMs, V. parahaemolyticus populations had decreased 0.493, 0.362, and 0.238 log10 CFU/g by the end of storage at 0, 4, and 10°C, respectively. Rates of reduction of V. parahaemolyticus shown in the TMs were 2.109, 1.579, and 0.894 log10 CFU/g for storage at 0, 4, and 10°C, respectively. Bacterial inactivation rates (IRs) estimated with the TMs (−0.245, −0.152, and −0.121 log10 CFU/day, respectively) were higher than those estimated with the MMs (−0.134, −0.0887, and −0.0732 log10 CFU/day, respectively) for storage at 0, 4, and 10°C. Higher viable V. parahaemolyticus numbers were predicted using the MMs than using the TMs. On the basis of this study, RNA-based predictive MMs are the more accurate and reliable models and can prevent false-negative results compared to TMs. IMPORTANCE One important method for validating postharvest techniques and for monitoring the behavior of V. parahaemolyticus is to establish predictive models. Unfortunately, previous predictive models established based on plate counting methods or on DNA-based PCR can underestimate or overestimate the number of surviving cells. This study developed and validated RNA-based molecular predictive models to describe the survival of V. parahaemolyticus in oysters during low-temperature storage (0, 4, and 10°C). The RNA-based predictive models show the advantage of being able to count all of the culturable, nonculturable, and stressed cells. By using primers targeting the tlh gene and pathogenesis-associated genes (tdh and trh), real-time RT-PCR can evaluate the total surviving V. parahaemolyticus population as well as differentiate the pathogenic ones from the total population. Reliable and accurate predictive models are very important for conducting risk assessment and management of pathogens in food.

Pitfalls in RET fusion detection using break-apart FISH probes in papillary thyroid carcinoma

2020 ◽  
Author(s):  
Yuanyuan Liu ◽  
Shafei Wu ◽  
Liangrui Zhou ◽  
Yong Guo ◽  
Xuan Zeng
Keyword(s):  
Papillary Thyroid Carcinoma ◽  
Thyroid Carcinoma ◽  
Reverse Transcription ◽  
False Positive ◽  
False Negative ◽  
Digital Pcr ◽  
Papillary Thyroid ◽  
Scoring Method ◽  
Reverse Transcription Pcr ◽  
Fish Cells

Abstract Objective A standardized procedure of fused REarranged during Transfection (RET) gene detection using fluorescence in situ hybridization (FISH) remains to be established in papillary thyroid carcinoma (PTC). Our purpose was to investigate false-negative and false-positive events and their FISH signal characteristics. Methods A total of 111 PTC cases were analyzed using break-apart FISH probe for RET status evaluation. All FISH results were validated using fusion-induced asymmetric transcription assay (FIATA)-based reverse transcription droplet digital PCR (RT-ddPCR). Then, suspected RET-positive cases were tested using quantitative reverse transcription-PCR (RT-qPCR) and following next-generation sequencing (NGS) for recognizing fusion variants. Results Thirty RET+ cases were revealed, including 20 CCDC6-RET (exon1;exon12), 6 NCOA4-RET (exon8;exon12), 1 NCOA4-RET (exon7;exon12), 1 CCDC186-RET (exon7;exon12), 1 ERC1-RET (exon12;exon12) and 1 SPECC1L-RET (exon9;exon12) tumors. All RET fusion cases occurred in the BRAF- population, with a prevalence of 41.7% (30/72). Four cases of 8–13% (cutoff was 7.6%) dominant isolated 3’ green (IG) FISH signals were RET-. One FISH- case with isolated 5’ red (IR) signals with 94% abnormal tumor cells was demonstrated to be positive, harboring the NCOA4-RET (exon7; exon12) variant. Compared to RET fusions characterized by dominant break-apart (BA) signals with 29–100% aberrant cells, RET + with dominant IG-signal patterns all showed more frequent FISH+ cells (84–92%). RET+ PTC with a BA signal pattern was more frequently found in unifocal lesions than in multifocal/bilateral tumors (p=0.049). Conclusions A false-positive or false-negative event may exist for RET status detection in PTCs using the traditional FISH scoring method with BA probes.

scholarly journals Applying Grounded Theory to Understand Asian Women’s Motivations to Donate Healthy Breast Tissue

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Cynthia Wu ◽  
Katherine E. Ridley-Merriweather ◽  
Anna Maria V. Storniolo
Keyword(s):  
Clinical Trials ◽  
Grounded Theory ◽  
Breast Tissue ◽  
Tissue Bank ◽  
Tissue Donation ◽  
Altruistic Behavior ◽  
Asian Identity ◽  
Future Work ◽  
Theoretical Saturation ◽  
Racial And Ethnic Minority

Background: The Komen Tissue Bank (KTB) is a clinical trial and the only global biorepository that collects and stores healthy breast tissue to be used as controls in breast cancer (BC) research. Due to a variety of barriers, there is a lack of participation by racial and ethnic minority women in tissue donation. In order to increase this participation, it is necessary to understand why or why not these populations choose to participate in clinical trials such as the KTB. This study used grounded theory methodology to explore the motivations behind Asian women’s decisions to donate their breast tissue.  Methods: Guided by grounded theory, we conducted interviews with previous breast tissue donors who self-identified as Asian (n=20). We then transcribed and coded the interviews to discover common attitudes and motivations for participating in breast tissue donation.  Findings: Preliminary findings were obtained from 11 interviews. We identified three common themes that influenced these women’s donations: altruistic behavior, comfort with science, and Asian identity. Identified sub-themes include factors such as personal ties to BC and background in research and clinical trials. It is of note that over half of the women expressed Asian identity and comfort with science as important factors, and all mentioned altruistic tendencies, either towards family or towards research and others. Conclusion and Future Work: We identified common factors for donating healthy breast tissue from using grounded theory to interview previous donors of Asian descent. We will transcribe and code 9 more interviews, as well as use those interviews to confirm theoretical saturation. The findings from this study will be used in the future to inform a framework for developing recruitment strategies to increase overall participation of historically excluded individuals in the KTB. Future work will include exploring the motivations of Latinas regarding donating their healthy breast tissue.

scholarly journals COVID-19 Test Positivity Rate as a marker for hospital overload

2021 ◽  
Author(s):  
Mauro Gaspari
Keyword(s):  
Health Care ◽  
Time Series ◽  
Strong Correlation ◽  
Molecular Tests ◽  
Screening And Surveillance ◽  
Italian Regions ◽  
Number Of Patients ◽  
Second Phases ◽  
Antigen Tests ◽  
The Impact

AbstractThe use of antigen tests for the diagnosis of COVID-19 in Italy has risen sharply in autumn 2020. Although, Italian regions like Alto Adige, Veneto, Toscana, Lazio, Piemonte and Marche did a large use of these tests for screening and surveillance purposes or for implementing diagnosis protocols, in addition to molecular tests, they were not reported in the statistics in the last months of 2020. As a consequence of this situation the test positivity rate (TPR) index, defined as the number of new positive cases divided by the number of tests, has lost in accuracy. Only in the recent days, starting from the 15th of January 2021, antigen tests have become part of the statistics for all the Italian regions. Despite the lack of data, we have noticed that TPR has a strong correlation with the number of patients admitted in hospitals, and that TPR peaks in general precede the peaks of hospitalized people which occur on average about 15 days later.In this paper, we have deepened this intuition, analysing the TPR course and its relationship with the number of hospitalized people. To conduct the study we have defined a novel version of the TPR index which takes into account the number of tests done with respect to the population (considering both molecular and antigen tests), the number of infected individuals, and the number of patients healed. Successively, starting from a limited set of data which were made available in November 2020, we have reconstructed the antigen tests time series of four Italian regions, and we computed the TPR index for them.The results show that TPR peaks precede peaks of hospitalized people in both the first and the second phases of the pandemic in Italy, provided that antigen tests are considered. Moreover, the TPR index trend, can be used to deduct important information on the course of the epidemic, and on the impact of COVID-19 in the health care system, which can be monitored in advance.

scholarly journals Testing Antigens, Antibodies, and Immune Cells in COVID-19 as a Public Health Topic—Experience and Outlines

2021 ◽  
Vol 18 (24) ◽  
pp. 13173
Author(s):  
Monica Neagu ◽  
Carolina Constantin ◽  
Mihaela Surcel
Keyword(s):  
Public Health ◽  
Immune Cells ◽  
Digital Pcr ◽  
Immune Memory ◽  
Diagnostic Strategies ◽  
Molecular Tests ◽  
Chemiluminescent Microparticle Immunoassay ◽  
Pcr Method ◽  
Antigen Testing ◽  
Research Domains

The current COVID-19 pandemic has triggered an accelerated pace in all research domains, including reliable diagnostics methodology. Molecular diagnostics of the virus and its presence in biological samples relies on the RT-PCR method, the most used and validated worldwide. Nonconventional tests with improved parameters that are in the development stages will be presented, such as droplet digital PCR or CRISPR-based assays. These molecular tests were followed by rapid antigen testing along with the development of antibody tests, whether based on ELISA platform or on a chemiluminescent microparticle immunoassay. Less-conventional methods of testing antibodies (e.g., lateral flow immunoassay) are presented as well. Left somewhere in the backstage of COVID-19 research, immune cells and, furthermore, immune memory cells, are gaining the spotlight, more so in the vaccination context. Recently, methodologies using flow-cytometry evaluate circulating immune cells in infected/recovered patients. The appearance of new virus variants has triggered a surge for tests improvement. As the pandemic has entered an ongoing or postvaccination era, all methodologies that are used to monitor public health focus on diagnostic strategies and this review points out where gaps should be filled in both clinical and research settings.

Veränderungen der extrazellulären Matrix in der Venenwand – Ursache der primären Varikosis?

VASA ◽  
2000 ◽  
Vol 29 (3) ◽  
pp. 173-177 ◽  
Author(s):  
Delia Kirsch ◽  
Dienes ◽  
Küchle ◽  
Duschner ◽  
Wahl ◽  
...  
Keyword(s):  
Matrix Protein ◽  
Statistical Evaluation ◽  
Varicose Veins ◽  
Confocal Laser ◽  
Vein Wall ◽  
Biological Investigation ◽  
Saphenous Veins ◽  
Tissue Specimens ◽  
Matrix Expression ◽  
Negative Controls

Background: Conflicting theories on the development of primary varicosis have led to the molecular biological investigation of the vein wall or, more accurately, of the extracellular matrix. It was the aim of this study to quantify matrix expression and to compare pathological changes in the vein wall with valve-orientated staging of varicosis, in order to determine indicators of the primary cause of varicosis. Materials and methods: Three hundred seventy-two tissue specimens of greater saphenous veins were obtained from 17 patients with varicosities and categorised according to Hach stage and procurement site. The specimens were compared with 36 specimens collected from six patients without varicosities, incubated with fluorescence-stained antibodies for collagen 4, laminin, fibronectin and tenascin prior to being assessed with confocal laser scan microscopy. In addition, 22 vein specimens (16 varicose, 6 normal veins) serving as negative controls were investigated. Results: Image analysis and statistical evaluation showed that compared with normal veins, varicose veins are associated with a significant increase in matrix protein expression for collagen 4, laminin and tenascin. A trend towards an increase in matrix expression was further observed for fibronectin. There was, however, no difference between varicose veins and clinically healthy vein segments inferior to a varicose segment. Conclusion: If the findings of the present investigation can be confirmed by other studies, alterations in the vein wall may be regarded as the primary cause of varicosis and valvular insufficiency as the result of these changes.

Cancer Patients' Attitudes toward Future Research Uses of Stored Human Biological Materials

2007 ◽  
Vol 2 (3) ◽  
pp. 15-22 ◽  
Author(s):  
Paul R. Helft ◽  
Victoria L. Champion ◽  
Rachael Eckles ◽  
Cynthia S. Johnson ◽  
Eric M. Meslin
Keyword(s):  
Cancer Patients ◽  
Tissue Bank ◽  
Mail Survey ◽  
Biological Materials ◽  
Survey Study ◽  
Cancer Center ◽  
Future Research ◽  
Policy Debate ◽  
Indiana University ◽  
Tissue Specimens

The policy debate concerning informed consent for future, unspecified research of stored human biological materials (HBM) would benefit from an understanding of the attitudes of individuals who contribute tissue specimens to HBM repositories. Cancer patients who contributed leftover tissue to the Indiana University Cancer Center Tissue Bank under such conditions were recruited for a mail survey study of their attitudes. Our findings suggest that a clear majority of subjects would permit unlimited future research on stored HBMs without re-contact and re-consent, and a significant minority appear to desire ongoing control over future research uses of their tissue. These differences merit further investigation and suggest that a policy of blanket consent for all future, unspecified research would be premature.

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