Standardization of Methods for Sampling the Distal Airspace in Mechanically Ventilated Patients using Heat Moisture Exchange Filter Fluid

2021 ◽  
Author(s):  
Julie A. Bastarache ◽  
J Brennan McNeil ◽  
Erin Plosa ◽  
Jennifer S. Sucre ◽  
V Eric Kerchberger ◽  
...  
Keyword(s):  
Total Protein ◽  
Dwell Time ◽  
Fluid Collection ◽  
Sample Collection ◽  
Moisture Exchange ◽  
Non Invasive ◽  
Fluid Analysis ◽  
Median Volume ◽  
Filter Type ◽  
Distal Airspace

Non-invasive sampling of the distal airspace in patients with Acute Respiratory Distress Syndrome (ARDS) has long eluded clinical and translational researchers. We recently reported that fluid collected from Heat Moisture Exchange filters (HME) closely mirrors fluid directly aspirated from the distal airspace. In the current study, we sought to determine fluid yield from different HME types, optimal HME circuit dwell time and reliability of HME fluid in reflecting the distal airspace. We studied fluid yield from 4 different filter types by loading increasing volumes of saline and measuring volume of fluid recovered. We collected filters after 1, 2 and 4 hours of dwell time for measurement of fluid volume and total protein from 13 subjects. After identifying 4 hours as the optimal dwell time, we measured total protein and IgM in HME fluid from 42 subjects with ARDS and 9 with hydrostatic pulmonary edema (HYDRO). We found that the fluid yield varies greatly by filter type. With timed sample collection, fluid recovery increased with increasing circuit dwell time with a median volume of 2.0 mL (IQR 1.2-2.7) after 4 hours. Total protein was higher in the 42 subjects with ARDS compared to 9 with HYDRO (median 708 µg/ml (IQR 244-2017) vs 364 µg/ml (IQR 136-578), p=0.047) confirming that total protein concentration in HME is higher in ARDS compared to hydrostatic edema. These studies establish a standardized HME fluid collection protocol and confirm that HME fluid analysis is a novel non-invasive tool for study of the distal airspace in ARDS.

scholarly journals Peritoneal fluid collection in healthy cattle and evaluation of changes in cell morphology during storage in refrigerated and non-refrigerated samples

2020 ◽  
Vol 40 (3) ◽  
pp. 158-164
Author(s):  
Nathali A.A. Sales ◽  
Karina K.M.C. Flaiban ◽  
Joandes H. Fonteque ◽  
Júlio A.N. Lisbôa
Keyword(s):  
Total Protein ◽  
Repeated Measures ◽  
Peritoneal Fluid ◽  
Protein Concentration ◽  
Room Temperature ◽  
Fluid Collection ◽  
Fluid Analysis ◽  
Healthy Cattle ◽  
The Right

ABSTRACT: This study aimed to evaluate the appropriate sites of abdominocentesis for peritoneal fluid collection in cattle and to investigate the time of cell viability in vitro, comparing three methods of sample conservation. Twenty-one healthy cattle (19 females and 2 males) were subjected to a laparocentesis procedure to obtain peritoneal fluid, with punctures in three defined sites: left cranial, right cranial, and right caudal. The total peritoneal fluid collected was divided into three aliquots and maintained under three preservation conditions: room temperature (26°C), refrigeration (4°C), and room temperature (26°C) with the addition of 1μL of 10% formaldehyde per 1mL of peritoneal fluid. The peritoneal fluid analysis performed immediately after collection consisted of: physical examination (color, appearance, volume, and specific gravity), biochemical measures (pH, total protein, fibrinogen, creatinine, and glucose), and cellularity (total and differential counts). The determination of proteins and the examination of cells were repeated in each separate aliquot at two, four, six, and eight hours after harvest. Data were analyzed through repeated measures ANOVA or Friedman test. The harvest was productive in 67% of cattle. The left cranial and the right cranial puncture sites were the most appropriate. Peritoneal fluid analyzed after collection, the total protein concentration ranged from 1.4 to 3.6g/dL, and number of leukocytes ranged from 54 to 1,322 cells/μL; 60 to 95% of leukocytes were lymphocytes. The protein concentration decreased, but the absolute values of leukocytes, lymphocytes, and segmented neutrophils did not change up to eight hours after collection, independent of the maintenance method. Cell lysis was delayed by cooling, and the addition of formaldehyde did not help preserve the integrity of cellular morphology. Laparocentesis is a safe and secure procedure in cattle and maybe more productive when performed in specific sites on the left or right sides of the cranial abdominal wall. Peritoneal fluid samples may be analyzed with reliable results for up to eight hours after collection when kept refrigerated and for up to six hours when kept at room temperature.

scholarly journals 13-gene DNA Methylation Analysis from Oral Brushing: A Promising Non Invasive Tool in the Follow-up of Oral Cancer Patients

2019 ◽  
Vol 8 (12) ◽  
pp. 2107 ◽  
Author(s):  
Davide B. Gissi ◽  
Achille Tarsitano ◽  
Andrea Gabusi ◽  
Roberto Rossi ◽  
Giuseppe Attardo ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Oral Cancer ◽  
Cancer Treatment ◽  
Sample Collection ◽  
Methylation Analysis ◽  
Non Invasive ◽  
Locoregional Relapse ◽  
Positive Score ◽  
Dna Methylation Analysis

Background: This study aimed to evaluate the prognostic value of a non-invasive sampling procedure based on 13-gene DNA methylation analysis in the follow-up of patients previously treated for oral squamous cell carcinoma (OSCC). Methods: The study population included 49 consecutive patients treated for OSCC. Oral brushing sample collection was performed at two different times: before any cancer treatment in the tumor mass and during patient follow-up almost 6 months after OSCC treatment, within the regenerative area after OSCC resection. Each sample was considered positive or negative in relation to a predefined cut-off value. Results: Before any cancer treatment, 47/49 specimens exceeded the score and were considered as positive. Six months after OSCC resection, 16/49 specimens also had positive scores in the samples collected from the regenerative area. During the follow-up period, 7/49 patients developed locoregional relapse: 6/7 patients had a positive score in the regenerative area after OSCC resection. The presence of a positive score after oral cancer treatment was the most powerful variable related to the appearance of locoregional relapse. Conclusion: 13-gene DNA methylation analysis by oral brushing may have a clinical application as a prognostic non-invasive tool in the follow-up of patients surgically treated for OSCC.

scholarly journals Factors influencing genotyping success and genotyping error rate of Eurasian otter (Lutra lutra) faeces collected in temperate Central Europe

2020 ◽  
Vol 67 (1) ◽  
Author(s):  
Marcia Sittenthaler ◽  
Eva Maria Schöll ◽  
Christoph Leeb ◽  
Elisabeth Haring ◽  
Rosemarie Parz-Gollner ◽  
...  
Keyword(s):  
Influential Factors ◽  
Lutra Lutra ◽  
Sample Treatment ◽  
Success Rates ◽  
Sample Collection ◽  
Allelic Dropout ◽  
Non Invasive ◽  
Eurasian Otter ◽  
Faecal Samples ◽  
Elusive Species

AbstractThe use of non-invasively collected DNA source material for genetic and genomic applications is usually characterized by low target DNA concentration and quality, genotyping errors and cost-intensive lab procedures. However, for otters (Lutrinae) as elusive species of conservation concern, genetic non-invasive sampling has become an important tool to study their ecology and demography. To increase cost-efficiency of monitoring programmes and to promote the expansion of genomic approaches to non-invasive samples, we aimed to refine sample collection and preparation. Therefore, we examined the effects of intrinsic sample characteristics (including diet), environmental conditions in the field and sample treatment in the molecular laboratory on the success of genotyping and allelic dropout (ADO) rates using microsatellite markers in 1970 fresh Eurasian otter (Lutra lutra) scats. Using fresh samples only, we probably eliminated one of the most important impediments of genotyping DNA from otter faecal samples beforehand. But, we observed higher genotyping success and lower ADO rates for anal glad secretions and faecal samples containing high proportions of mucus. Moist conditions during sample collection may promote DNA degradation and PCR inhibition, leading to decreased genotyping success rates. ADO was further affected by the type of extraction kit. However, a high proportion of variance remaining unexplained by our models implied that additional parameters were acting (amount of PCR inhibitors, non-uniform distribution of intestinal cells, efficiency of PCRs, specific microclimate at marking sites). We summarized influential factors maximizing genotyping quality of otter scats and give recommendations for sample collection, storage and DNA extraction based on our results and current literature.

scholarly journals Saliva Samples as A Tool to Study the Effect of Meal Timing on Metabolic And Inflammatory Biomarkers

Nutrients ◽  
2020 ◽  
Vol 12 (2) ◽  
pp. 340 ◽  
Author(s):  
Katharina Kessler ◽  
Silke Hornemann ◽  
Natalia Rudovich ◽  
Daniela Weber ◽  
Tilman Grune ◽  
...  
Keyword(s):  
Metabolic Regulation ◽  
Diurnal Variations ◽  
Inflammatory Biomarkers ◽  
Blood Levels ◽  
Fat Intake ◽  
Sample Collection ◽  
High Fat ◽  
Non Invasive ◽  
Promising Tool ◽  
Meal Timing

Meal timing affects metabolic regulation in humans. Most studies use blood samples for their investigations. Saliva, although easily available and non-invasive, seems to be rarely used for chrononutritional studies. In this pilot study, we tested if saliva samples could be used to study the effect of timing of carbohydrate and fat intake on metabolic rhythms. In this cross-over trial, 29 nonobese men were randomized to two isocaloric 4-week diets: (1) carbohydrate-rich meals until 13:30 and high-fat meals between 16:30 and 22:00 or (2) the inverse order of meals. Stimulated saliva samples were collected every 4 h for 24 h at the end of each intervention, and levels of hormones and inflammatory biomarkers were assessed in saliva and blood. Cortisol, melatonin, resistin, adiponectin, interleukin-6 and MCP-1 demonstrated distinct diurnal variations, mirroring daytime reports in blood and showing significant correlations with blood levels. The rhythm patterns were similar for both diets, indicating that timing of carbohydrate and fat intake has a minimal effect on metabolic and inflammatory biomarkers in saliva. Our study revealed that saliva is a promising tool for the non-invasive assessment of metabolic rhythms in chrononutritional studies, but standardisation of sample collection is needed in out-of-lab studies.

scholarly journals Non-invasive sample collection for respiratory virus testing by multiplex PCR

2011 ◽  
Vol 52 (3) ◽  
pp. 210-214 ◽  
Author(s):  
Anne J. Blaschke ◽  
Mandy A. Allison ◽  
Lindsay Meyers ◽  
Margarita Rogatcheva ◽  
Caroline Heyrend ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Respiratory Virus ◽  
Sample Collection ◽  
Non Invasive ◽  
Virus Testing

scholarly journals COVID-19 salivary signature: diagnostic and research opportunities

2020 ◽  
pp. jclinpath-2020-206834 ◽  
Author(s):  
Dipak Sapkota ◽  
Tine Merete Søland ◽  
Hilde Kanli Galtung ◽  
Lars Peter Sand ◽  
Simone Giannecchini ◽  
...  
Keyword(s):  
Disease Transmission ◽  
Scientific Basis ◽  
Healthcare Personnel ◽  
Sample Collection ◽  
Promising Alternative ◽  
Invasive Approach ◽  
Non Invasive ◽  
Saliva Collection ◽  
Personnel Safety ◽  
Degrees Of Severity

The COVID-19 (caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)) epidemic started in Wuhan (Hubei Province, China) in mid-December 2019 and quickly spread across the world as a pandemic. As a key to tracing the disease and to implement strategies aimed at breaking the chain of disease transmission, extensive testing for SARS-CoV-2 was suggested. Although nasopharyngeal/oropharyngeal swabs are the most commonly used biological samples for SARS-CoV-2 diagnosis, they have a number of limitations related to sample collection and healthcare personnel safety. In this context, saliva is emerging as a promising alternative to nasopharyngeal/oropharyngeal swabs for COVID-19 diagnosis and monitoring. Saliva collection, being a non-invasive approach with possibility for self-collection, circumvents to a great extent the limitations associated with the use of nasopharyngeal/oropharyngeal swabs. In addition, various salivary biomarkers including the salivary metabolomics offer a high promise to be useful for better understanding of COVID-19 and possibly in the identification of patients with various degrees of severity, including asymptomatic carriers. This review summarises the clinical and scientific basis for the potential use of saliva for COVID-19 diagnosis and disease monitoring. Additionally, we discuss saliva-based biomarkers and their potential clinical and research applications related to COVID-19.

scholarly journals Vascular Endothelial Growth Factor in Tear Samples of Patients with Systemic Sclerosis

2015 ◽  
Vol 2015 ◽  
pp. 1-4 ◽  
Author(s):  
Anikó Rentka ◽  
Jolán Hársfalvi ◽  
András Berta ◽  
Krisztina Köröskényi ◽  
Zoltán Szekanecz ◽  
...  
Keyword(s):  
Systemic Sclerosis ◽  
Total Protein ◽  
Protein Production ◽  
Average Concentration ◽  
Vascular Endothelial ◽  
Ophthalmological Examination ◽  
Sample Collection ◽  
Total Protein Level ◽  
And Control ◽  
Endothelial Growth Factor

Background. Systemic sclerosis is an autoimmune disease, characterized by widespread small vessel vasculopathy, immune dysregulation with production of autoantibodies, and progressive fibrosis. Changes in levels of proangiogenic cytokines had already been determined largely in serum. Our aim was to assess the levels of VEGF in human tears of patients with SSC.Patients and methods. Forty-three patients (40 female and 3 men, mean (SD) age 61 (48–74) years) with SSc and 27 healthy controls were enrolled in this study. Basal tear sample collection and tear velocity investigations were carried out followed by an ophthalmological examination. Total protein concentrations and VEGF levels were determined in tear samples.Results. The average collected tear fluid volume developed 10.4 μL (1.6–31.2) in patients and 15.63 μL (3.68–34.5) in control subjects. The average total protein level was 6.9 μg/μL (1.8–12.3) in tears of patients and control tears contained an average of 4.132 μg/μL (0.1–14.1) protein. In patients with SSc the average concentration of VEGF was 4.9 pg/μL (3.5–8.1) and 6.15 pg/μL (3.84–12.3) in healthy samples.Conclusions. Total protein production was increased because of the smaller tear volume. Decreased VEGF in tear of SSc patients can be explained also by the decreased tear secretion of patients.

scholarly journals Opportunities and challenges in metabarcoding approaches for helminth community identification in wild mammals

Parasitology ◽  
2017 ◽  
Vol 145 (5) ◽  
pp. 608-621 ◽  
Author(s):  
TUOMAS AIVELO ◽  
ALAN MEDLAR
Keyword(s):  
Statistical Analysis ◽  
Bacterial Community ◽  
Bacterial Community Composition ◽  
Accurate Assessment ◽  
Helminth Community ◽  
Sample Collection ◽  
Wild Mammals ◽  
Non Invasive ◽  
Community Identification ◽  
Parasitological Study

SUMMARYDespite metabarcoding being widely used to analyse bacterial community composition, its application in parasitological research remains limited. What interest there has been has focused on previously intractable research settings where traditional methods are inappropriate, for example, in longitudinal studies and studies involving endangered species. In settings such as these, non-invasive sampling combined with metabarcoding can provide a fast and accurate assessment of component communities. In this paper we review the use of metabarcoding in the study of helminth communities in wild mammals, outlining the necessary procedures from sample collection to statistical analysis. We highlight the limitations of the metabarcoding approach and speculate on what type of parasitological study would benefit from such methods in the future.

scholarly journals A robotic platform for fluidically-linked human body-on-chips experimentation

2019 ◽  
Author(s):  
Richard Novak ◽  
Miles Ingram ◽  
Susan Clauson ◽  
Debarun Das ◽  
Aaron Delahanty ◽  
...  
Keyword(s):  
Human Body ◽  
Experimental System ◽  
Blood Substitute ◽  
Sample Collection ◽  
Human Organ ◽  
Non Invasive ◽  
Organ Specific ◽  
Repeated Sampling ◽  
Substitute Medium

Here we describe of an ‘Interrogator’ instrument that uses liquid-handling robotics, a custom software package, and an integrated mobile microscope to enable automated culture, perfusion, medium addition, fluidic linking, sample collection, andin situmicroscopic imaging of up to 10 Organ Chips inside a standard tissue culture incubator. The automated Interrogator platform maintained the viability and organ-specific functions of 8 different vascularized, 2-channel, Organ Chips (intestine, liver, kidney, heart, lung, skin, blood-brain barrier (BBB), and brain) for 3 weeks in culture when fluidically coupled through their endothelium-lined vascular channels using a common blood substitute medium. When an inulin tracer was perfused through the multi-organ Human Body-on-Chips (HuBoC) fluidic network, quantitative distributions of this tracer could be accurately predicted using a physiologically-based multi-compartmental reduced order (MCRO)in silicomodel of the experimental system derived from first principles. This automated culture platform enables non-invasive imaging of cells within human Organ Chips and repeated sampling of both the vascular and interstitial compartments without compromising fluidic coupling, which should facilitate future HuBoc studies and pharmacokinetics (PK) analysisin vitro.

Sign in / Sign up

Export Citation Format

Share Document