Optimizing tissue stewardship in non-small cell lung cancer to support molecular characterization and treatment selection: statement from a working group of thoracic pathologists.

Many patients with non-small cell lung cancer do not receive guideline-recommended, biomarker-directed therapy, despite the potential for improved clinical outcomes. Access to timely, accurate, and comprehensive molecular profiling, including targetable protein overexpression, is essential to allow fully informed treatment decisions to be taken. In turn, this requires optimal tissue management to protect and maximize the use of this precious finite resource. Here, a group of leading thoracic pathologists recommend factors to consider for optimal tissue management. Starting from when lung cancer is first suspected, keeping predictive biomarker testing in the front of the mind should drive the development of practices and procedures that conserve tissue appropriately to support molecular characterization and treatment selection.

Quality, acceptability and usability of self-sampling kits used by non-healthcare professionals for STI diagnosis in Spain: a single-blind study.

OBJECTIVES: Sexually transmitted infections (STIs) have markedly increased over the last decade in Spain, calling for prevention and control innovative approaches. While there is evidence indicating the effectiveness of self-sampling for STI diagnosis, no kits for this purpose have been authorised in Spain. METHODS: A prospective single-blind cross-sectional study carried out between November and December 2022 in an STI clinic in Madrid, Spain, to determine the validity, feasibility and acceptability of self-sampling kits used by non-healthcare professionals from vagina, pharynx, rectum and urethra to diagnose Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG). Self-samples were compared with samples collected by healthcare professional (HC samples) and analysed by PCR. Frequency of CT and NG diagnosis by sample type was compared using McNemar's test for paired data. Sensitivity and specificity of self-samples for CT and NG diagnosis were also calculated. RESULTS: 306 self-samples from 51 participants were analysed. 80% were men with median age of 33 (IQR: 28-38) years. Self-samples and HC samples showed no significant statistical differences in CT and NG diagnosis. Self-samples had a sensitivity of 81% for CT and 93% for NG, with a specificity of 97% for CT and 95% for NG. More than 90% of participants had no difficulty understanding the kit instructions and 71% expressed high levels of satisfaction with the self-sampling kit. CONCLUSION: Self-sampling kits for CT and NG diagnosis can be safely and effectively used by non-healthcare professionals in Spain. National strategies for STI prevention and control should prioritise self-sampling strategies.

Extensive comparison of salivary collection, transportation, preparation, and storage methods: a systematic review.

BACKGROUND: Human saliva as a bodily fluid-similar to blood-is utilized for diagnostic purposes. Unlike blood sampling, collecting saliva is non-invasive, inexpensive, and readily accessible. There are no previously published systematic reviews regarding different collection, transportation, preparation, and storage methods for human saliva. DESIGN: This study has been prepared and organized according to the preferred reporting items for systematic reviews and meta-analyses (PRISMA) 2020 guidelines. This systematic review has been registered at PROSPERO (Registration ID: CRD42023415384). The study question according to the PICO format was as followed: Comparison of the performance (C) of different saliva sampling, handling, transportation, and storage techniques and methods (I) assessed for analyzing stimulated or unstimulated human saliva (P and O). An electronic search was executed in Scopus, Google Scholar, and PubMed. RESULTS: Twenty-three descriptive human clinical studies published between 1995 and 2022 were included. Eight categories of salivary features and biomarkers were investigated (i.e., salivary flow rate, total saliva quantity, total protein, cortisol, testosterone, DNA quality and quantity, pH and buffering pH). Twenty-two saliva sampling methods/devices were utilized. Passive drooling, Salivette®, and spitting were the most utilized methods. Sampling times with optimum capabilities for cortisol, iodine, and oral cancer metabolites are suggested to be 7:30 AM to 9:00 AM, 10:30 AM to 11:00 AM, and 14:00 PM to 20:00 PM, respectively. There were 6 storage methods. Centrifuging samples and storing them at -70 °C to -80 °C was the most utilized storage method. For DNA quantity and quality, analyzing samples immediately after collection without centrifuging or storage, outperformed centrifuging samples and storing them at -70 °C to -80 °C. Non-coated Salivette® was the most successful method/device for analyzing salivary flow rate. CONCLUSION: It is highly suggested that scientists take aid from the reported categorized outcomes, and design their study questions based on the current voids for each method/device.

Detecting human cytomegalovirus in urine, vagina and saliva: Impact of biological fluids and storage durations and temperatures on CMV DNA recovery.

Sample collection, transport and storage conditions vary in the human cytomegalovirus (CMV) shedding literature. Currently, limited data exist on the impact of biological fluids and pre-analytical sample handling on the detection of CMV DNA. To evaluate CMV DNA recovery from urine, vaginal fluid and saliva stored in different conditions, adult urine, vaginal and saliva fluids and swabs, stored with or without selected nucleic acid preservation media at various durations and temperatures, was compared by polymerase chain reaction (PCR) quantitation of spiked samples and self-collected urine (n = 45) and vaginal swabs (n = 58) from CMV seropositive pregnant women. There was a time-dependent reduction in CMV DNA recovery from urine, urine diluted in phosphate-buffered saline, and saliva stored at 2-8°C, but not from urine preserved in cobas® PCR transport media (CPM) (urine/CPM). For vaginal fluid, a reduction in recovery was evident after 7 days storage at 2-8°C. CMV DNA recovery over 91 days was similar between -80°C and -20°C storage for urine and vaginal swabs preserved in CPM, and saliva swabs preserved in eNAT® PCR transport media. A statistically significant change in CMV DNA recovery after 25 months storage (median) at -80°C was not observed for self-collected urine/CPM and vaginal swab/CPM from pregnant women. Taken together, recovery of CMV DNA is dependent on fluid type and storage conditions. To improve the validity and reliability of detection at different storage durations and temperatures, the use of nucleic acid preserving transport media at the point of collection for urine, vaginal fluid and saliva may be essential.

Environmental contamination by Chlamydia trachomatis and Neisseria gonorrhoeae: is it time to change our infection control practices? Results of a regional study.

OBJECTIVES: Nucleic acid amplification tests (NAATs) are highly sensitive for the detection of Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) DNA/ribosomal RNA (rRNA). Previous studies have demonstrated contamination of surfaces in sexual health clinics (SHCs) with CT/NG. False positive results can occur if patient samples are contaminated by environmental DNA/rRNA. This can have a dramatic impact on patients' lives and relationships. Previous attempts to reduce contamination, through staff training alone, have been unsuccessful. We aimed to investigate environmental contamination levels in SHCs and to assess a two-armed intervention aimed at reducing surface contamination. METHODS: Questionnaires were sent to 10 SHCs. Six clinics, with differing characteristics, were selected to participate in sample collection. Each clinic followed standardised instructions to sample surfaces using a CT/NG NAAT swab. Clinics were invited to introduce the two-armed intervention. The first arm was cleaning with a chlorine-based cleaning solution once daily. The second arm involved introducing clinic-specific changes to reduce contamination. RESULTS: 7/10 (70%) clinics completed the questionnaire. Overall, 88/263 (33%) swabs were positive for CT/NG. Clinics 1, 3 and 4 had high levels of contamination, with 28/64 (44%), 17/33 (52%) and 30/52 (58%) swabs testing positive, respectively. Clinics 2 and 6 had lower levels of contamination, with 7/46 (15%) and 6/35 (17%), respectively. 0/33 (0%) of swabs were positive at clinic 5 and this was the only clinic already using a chlorine-based solution to clean all surfaces and delivering twice-yearly clinic-specific infection control training. Following both intervention arms at clinic 1, 2/50 (4%, p<0.0001) swabs tested positive for CT/NG. Clinic 4 introduced each arm separately. After the first intervention, 13/52 (25%, p=0.003) swabs tested positive and following the second arm 4/50 (8%, p<0.0001) swabs were positive. CONCLUSIONS: Environmental contamination is a concern in SHCs. We recommend that all SHCs monitor contamination levels and, if necessary, consider using chlorine-based cleaning products and introduce clinic-specific changes to address environmental contamination.

Systematic orientation of fresh rectal suction biopsies improves histopathological diagnostics in hirschsprung's disease - a method description and preliminary report.

BACKGROUND: Optimizing rectal suction biopsy (RSB) diagnostics in Hirschsprung's disease (HD) may shorten diagnostic time and prevent need for repeated biopsies. AIM: To explore whether systematic orientation of fresh RSB specimens increased biopsy quality, diagnostic times, diagnostic efficacy, and histopathologic workload, and to explore these outcome measures for aganglionic specimens. MATERIALS/METHODS: This was an observational case-control study conducted at a national referral center for HD on data collected from the local HD-diagnostic register. From 2019 each fresh RSB was oriented by the collector in a notch in a foam cushion, placed in a separate cassette, and sent in formalin for pathological analysis. Outcome measures of oriented RSB samples collected 2019-2021 were compared to those of non-oriented RSB samples collected 2015-2018. Staining/immunohistochemistry consisted of hematoxylin eosin, S-100 and calretinin. RESULTS: 78 children with 81 RSBs and 242 biopsy analyzes were included. The frequency of high-quality RSB specimens was higher in oriented: 40% (42/106) versus non-oriented 25% (34/136) (p = 0.018), the diagnostic turnaround time was shorter: 2 days (1-5) versus 3 days (2-8) (p = 0.015), and the number of additional sectioning/leveling/re-orientation per biopsy was lower: 7 (3-26) versus 16 (7-72) (p = 0.011). Specifically for aganglionic specimens, the frequency of high-quality biopsies was generally higher in oriented than in non-oriented RSB specimens: 47% (28/59) versus 14% (7/50) (p < 0.001); the diagnostic efficacy was higher 95% (19/20) versus 60% (9/15) (p = 0.027) and the diagnostic turnaround time shorter: 2 days (2-3) versus 3 days (2-8) (p = 0.036). CONCLUSIONS: Systematic orientation of fresh RSB specimens improves HD diagnostics. Improvement was consistent in aganglionic specimens.

At-home sample collection is an effective strategy for diagnosis and management of symptomatic and asymptomatic SARS-CoV-2 carriers.

BACKGROUND: Diagnostic testing accessibility and asymptomatic transmission of SARS-CoV-2 present major challenges for curbing and preventing community prevalence of COVID-19. At-home sample collection for molecular testing provides a convenient and effective solution for disease containment and prevention. METHODS: This is a retrospective, cross-sectional, case-control study. Our primary aim was to determine the prevalence and relative risk of SARS-CoV-2 among asymptomatic versus symptomatic individuals using at-home sample collection kits for diagnosis. Participants included adults from across the United States who completed a COVID-19 Home Collection kit between May 2020 and September 2021. Main measurements included self-reported symptoms and at-home self-collected anterior nasal swab RT-PCR test results for SARS-CoV-2. RESULTS: Data from 282,831 individuals were included in this analysis. The overall SARS-CoV-2 prevalence of at-home test takers was low compared to national averages during this period (3.28% vs. 7.68%). Those reporting no symptoms were at lower risk of positive test results compared to those with symptoms (risk ratio: 0.080, 95% CI, 0.078-0.082). However, of all positive SARS-CoV-2 tests, 48.75% were from individuals reporting no symptoms at the time of testing. CONCLUSIONS: We conclude that at-home sample collection is a viable option and potentially important strategy for improving access to testing, detecting asymptomatic cases, and curbing preventable transmission of COVID-19.

Preclinical coronavirus studies and pathology: Challenges of the high-containment laboratory.

The COVID-19 pandemic has highlighted the critical role that animal models play in elucidating the pathogenesis of emerging diseases and rapidly analyzing potential medical countermeasures. Relevant pathologic outcomes are paramount in evaluating preclinical models and therapeutic outcomes and require careful advance planning. While there are numerous guidelines for attaining high-quality pathology specimens in routine animal studies, preclinical studies using coronaviruses are often conducted under biosafety level-3 (BSL3) conditions, which pose unique challenges and technical limitations. In such settings, rather than foregoing pathologic outcomes because of the inherent constraints of high-containment laboratory protocols, modifications can be made to conventional best practices of specimen collection. Particularly for those unfamiliar with working in a high-containment laboratory, the authors describe the logistics of conducting such work, focusing on animal experiments in BSL3 conditions. To promote scientific rigor and reproducibility and maximize the value of animal use, the authors provide specific points to be considered before, during, and following a high-containment animal study. The authors provide procedural modifications for attaining good quality pathologic assessment of the mouse lung, central nervous system, and blood specimens under high-containment conditions while being conscientious to maximize animal use for other concurrent assays.

A Novel Cryogenic Adhesive Retaining Fluidity at Dry-Ice Temperature for Low-Temperature Scanning Electron Microscopy.

Scanning electron microscopy operated at cryogenic temperature (cryo-SEM) is a powerful tool for investigating surface and cross-sectional nanostructures of water-containing samples. Typically, cryo-SEM samples are frozen just before observation in specific metal carriers. However, pre-frozen samples are also of interest, such as frozen food and freeze-stored animal samples. In such cases, sample mounting with a defined orientation is required, but there has been a lack of ideal conductive adhesives that can be used without increasing the sample temperature. Here, we developed a mixture of graphite oxide and 1,3-butanediol as an adhesive, capable of gluing samples at dry-ice temperature and is frozen below that temperature. Dispersion of graphite oxide increased the conductivity and reduced the charge-up contrast. Acquisition of energy-dispersive X-ray spectrum, cross-sectional ion milling, and high-resolution imaging were successfully achieved using the adhesive. We tested and confirmed the usefulness of this new adhesive by applying it to cryo-SEM surface imaging of diatomite, freeze-fractured cross-sectional imaging of chicken liver, and ion milling cross-sectional imaging of a deep-sea snail. The new adhesive is not only useful for food science and field-preserved biological samples but also potentially applicable to wider fields such as archaeological and biological samples preserved under permafrost.

Efficacy of topical steroids for the treatment of olfactory disorders caused by COVID-19: A systematic review and meta-analysis.

OBJECTIVES: The aim of this study was to assess the effect of topical steroids on acute-onset olfactory dysfunction in patients infected with COVID-19. DESIGN AND SETTING: Systematic review and meta-analysis of cohort studies. PARTICIPANTS: Patients infected with COVID-19. MAIN OUTCOME MEASURES: PubMed, Embase, the Web of Science, SCOPUS, Cochrane database and Google Scholar were searched for articles up to September 2021. We analysed studies comparing the improvement of olfactory dysfunction between topical steroid treatment and control groups (placebo or no treatment). In addition, we performed a subgroup analysis by study type. RESULTS: The improvement of olfactory score at 2 (standardised mean difference [SMD] = 0.7272, 95% confidence interval = [0.3851, 1.0692], p < .0001, I2  = 62.1%) and 4 weeks post-treatment (SMD = 1.0440 [0.6777, 1.4102], p < .0001, I2  = 61.2%) was statistically greater in the treatment than control group. However, there was no significant difference (odds ratio [OR] = 1.4345 [0.9525, 2.1604], p = .0842, I2  = 45.4%) in the incidence of fully recovery from anosmia/hyposmia between the treatment and control groups. In subgroup analysis, there were no significant differences in the improvement of olfactory score at 4 weeks post-treatment (OR = 0.6177 [0.1309, 1.1045] vs. 0.1720 [0.8002, 1.5438], p = .0761) or the incidence of full recovery from anosmia/hyposmia (OR = 1.8478 [0.6092, 5.6053] vs. 1.3784 [0.8872, 2.1414], p = .8038) between randomised and non-randomised controlled trials. CONCLUSIONS: Although this meta-analysis found that topical steroids improved the acute-onset olfactory dysfunction caused by COVID-19, there was no difference in the rate of full olfactory recovery between treated and control patients.

Flow cytometric evaluation of platelet-leukocyte conjugate stability over time: methodological implications of sample handling and processing.

Platelet activation and subsequent aggregation is a vital component of atherothrombosis resulting in acute myocardial infarction. Therefore, quantifying platelet aggregation is a valuable measure for elucidating the pathogenesis of acute coronary syndromes (ACS). Circulating platelet-monocyte conjugates (PMC) as determined by flow cytometry (FCM) are an important measure of in vivo platelet aggregation. However, the influence of sample handling on FCM measurement of PMC is not well-studied. The changes in FCM measurement of PMC with variation in sample handling techniques were evaluated. The stability of PMC concentrations over time with changes in fixation and immunolabeling intervals was assessed. The effect of Time-to-Fix and Time-to-Stain on FCM PMC measurements was investigated in five healthy volunteers. Time-to-Fix (i.e., interval between phlebotomy and sample fixation) was performed at 3, 30, and 60 min. Time-to-Stain (i.e., time of fixed sample storage to staining) was performed at 1, 24, and 48 h. Increasing Time-to-Stain from 1 to 24 or 48 h resulted in lower PMC measures (p < 0.0001). A statistically significant difference in PMC measurement with increasing Time-to-Fix was not observed (p < 0.41). Postponement of sample staining has deleterious effects on the measurement of PMC via FCM. Delays in immunolabeling of fixed samples compromised measurement of PMC by 30% over the first 24 h. Staining/FCM should be completed within an hour of collection.

Effects of a pneumatic tube system on the hemolysis of blood samples: a PRISMA-compliant meta-analysis.

Many studies have explored how using a pneumatic tube system (PTS) is related to the hemolysis of blood samples, but their conclusions have been inconsistent. This meta-analysis was to clarify whether using a PTS induces the hemolysis of blood samples. The PubMed, Embase, Scopus, CNKI, CqVip, SinoMed and WanFang databases were searched for studies published between January 1970 and August 2019. The primary outcomes were the hemolysis rate and hemolysis index of blood samples after applying a PTS and manual transportation. We estimated the pooled risk ratio (RR) and the standardized mean difference (SMD), using random-effects models. This meta-analysis included 29 studies covering 3121 blood samples. No significant differences were found between the PTS and manual-transportation groups in the hemolysis rate [RR: 0.99, 95% confidence interval (CI): 0.57 to 1.70], hemolysis index (SMD: 0.19, 95% CI: -0.00 to 0.38), or level of potassium (SMD: 0.05, 95% CI: -0.03 to 0.12), alanine aminotransferase (SMD: 0.00, 95% CI: -0.10 to 0.11), or aspartate aminotransferase (SMD: 0.04, 95% CI: -0.08 to 0.17). However, lactate dehydrogenase (LDH) level was significantly higher in the PTS group than in the manual-transportation group (SMD: 0.20, 95% CI: 0.06 to 0.34). Subgroup analysis revealed that the LDH level was clearly higher in the PTS group than in the manual-transportation group only when the PTS speed was ≥6 m/s or when the PTS distance was ≥250 m. According to this meta-analysis, PTSs were associated with alterations in LDH measurements, so it is sensible that each hospital validates and monitors their PTSs.

Rapid onsite evaluation uses a very small proportion of total material aspirated at endobronchial or endoscopic ultrasound in the investigation of suspected thoracic malignancy.

INTRODUCTION: The objectives were: to measure the proportion of aspirated material used to make direct slides for rapid onsite evaluation (ROSE) at endobronchial (EBUS) and endoscopic ultrasound (EUS) in suspected thoracic malignancy; and to correlate pass weights with ROSE category and needle size. METHOD: All EBUS and EUS cases for possible thoracic malignancy October 2018-May 2019 were included. All material from each pass was expelled into a Petri dish. One drop of material was placed on each of two slides; one used for ROSE, the other fixed and remaining material processed to cell block. Dish and slides were weighed before and after this procedure on a sensitive balance and weight of aspirate and slide material calculated. When ROSE identified malignancy, slide production ceased but target sampling for ancillary studies continued. RESULTS: ROSE accuracy was 96.8%. Mean percentage by target of aspirated material used to make direct slides for ROSE was 1.9% in malignant cases and 3.6% in non-malignant cases (P = .027 for difference). Mean percentage by pass was 5.9%. Mean weight of a single aspirate was 128.8 mg. Mean weight of aspirates insufficient on ROSE (175.7 mg) was significantly higher than the mean weight of benign or malignant aspirates (117.1 and 114.0 mg, respectively). Mean weight of aspirates using 22G needles (132.6 mg) was significantly higher than that for 25G needles (87.1 mg). CONCLUSION: Material made into direct slides at EBUS and EUS and used in part for ROSE uses a tiny proportion of aspirated material with over 98% processed to cell block and available for ancillary testing in malignant cases.

A comparison of swab types on sample adequacy, suspects comfort and provider preference in COVID-19.

AIM: This study was aimed to compare the virological, suspect reported outcomes and provider preferences during COVID-19 swab taking procedure used for sampling. METHODS: The COVID-19 suspects are subjected to nasopharyngeal (NP) and oropharyngeal (OP) swabs for testing. Two types of swabs (Nylon and Dacron) are used for sample collection. Prospectively each suspect's response is collected and assessed for self-reported comfort level. The provider's experience with each suspect and virological outcomes recorded separately. The sample adequacy was compared based on swab types and demographic characteristics. RESULTS: A total of 1008 COVID-19 suspects were considered for comparison of various outcomes. Dacron and flocked Nylon swab sticks are used for taking 530 and 478 samples, respectively. Suspects who underwent the procedure using Nylon swabs were six times more likely to have pain/discomfort compared to when Dacron swab was used (Adj RR (95% CI: 6.76 (3.53 to 13, p=0.0001))). The providers perceived six times more resistance with the Nylon swabs compared to Dacron Swabs (Adj RR (95% CI: 5.96 (3.88 to 9.14, p=0.0001))). The pediatric population had a higher rate of blood staining in Dacron swab [Dacron 66 (80.5%); Nylon 51 (54.8%) p=0.0001]. The sample adequacy rate and laboratory positivity rate were not significantly different from each other. CONCLUSIONS: Given the comparable virological outcomes, the difference in suspect and providers comfort should drive swab selection based on characteristics of the suspects. The bulbous Nylon swab caused more pain/discomfort in adults compared to Dacron.

Insufficient mixing of thawed serum samples leading to erroneous results - experience from a field study and use of a correction procedure.

Incorrect analysis results that are close to expected might not be recognized in scientific studies or routine patient care. In two field studies we obtained unexpected results in a large number of samples. The present study aimed to identify the source of error in the samples from these studies and to validate a method to obtain correct results. Pre-analytical procedures were scrutinized, giving no indications of inappropriate pre-analytical sample handling in the field or during transport in a tropical climate. Using a new set of samples from volunteers in simulation experiments, we observed the known concentration gradient of analytes sampled in gel as well as plain tubes after freezer storage and thawing. Experiments demonstrated that mixing of samples by vortexing alone was not sufficient to disrupt the gradient formed by freezing and thawing, which appeared to cause the problem encountered when we in field studies analyzed and biobanked large sample sets by robot pipetting. A correction procedure was introduced, in which the obtained value of an analyte was multiplied by a correction factor calculated for each sample using the expected sodium level (140 mmol/L) divided by the measured sodium value. When it was validated on results from the simulation experiments, we repeatedly found that the correction lead to results very close to true values for analytes of different size and charge. Usefulness of the procedure was demonstrated when applied to a large set of field study results.

Heparin and citrate additive carryover during blood collection.

Background Published evidence on the risk of additive carryover during phlebotomy remains elusive. We aimed to assess potential carryover of citrated and heparinized blood and the relative volume needed to bias clinical chemistry and coagulation tests. Methods We simulated standardized phlebotomies to quantify the risk of carryover of citrate and heparin additives in distilled water, using sodium and lithium as surrogates. We also investigated the effects of contamination of heparinized blood samples with increasing volumes of citrated blood and pure citrate on measurements of sodium, potassium, chloride, magnesium, total and ionized calcium and phosphate. Likewise, we studied the effects of contamination of citrated blood samples with increasing volumes of heparinized blood on heparin (anti-Xa) activity, lithium, activated partial thromboplastin time (APTT), prothrombin time (PT) and thrombin time (TT). We interpreted these results based on measurement deviations beyond analytical, biological and clinical significance. Results Standardized phlebotomy simulations revealed no significant differences in concentration of surrogate markers. Clinically significant alterations were observed after contamination of heparinized blood samples with volumes of citrated blood beyond 5-50 µL for ionized calcium and beyond 100-1000 µL for sodium, chloride and total calcium. Investigations of pure citrate carryover revealed similar results at somewhat lower volumes. Heparinized blood carryover showed clinically significant interference of coagulation testing at volumes beyond 5-100 µL. Conclusions Our results suggest that during a standardized phlebotomy, heparin or citrate contamination is highly unlikely. However, smaller volumes are sufficient to severely alter test results when deviating from phlebotomy guidelines.

Sampling of urine for diagnosing urinary tract infection in general practice - First-void or mid-stream urine?

OBJECTIVE: To study 1) whether the accuracy of point-of-care (POC) urine tests (dipstick, phase-contrast microscopy and culture) differs when performed on first-void urine (FVU) compared to mid-stream urine (MSU), and 2) if a delay of analysis up to four hours affects the accuracy of POC tests when performed on urine from symptomatic of urinary tract infection (UTI), non-pregnant women in general practice. DESIGN: Prospective diagnostic study using paired samples. Setting/Intervention: Three general practices in Copenhagen. Each woman delivered FVU and MSU samples from the same void. As a reference standard, 8 ml of MSU was sent for culture at the Microbiology Department. PATIENTS: 117 women with one or more symptoms of UTI. MAIN OUTCOME MEASURES: Sensitivity, specificity and agreement with the reference standard of FVU and MSU with different time delays (zero vs. one vs. four hours) as compared to reference standard (MSU at time zero in boric acid tubes). RESULTS: All three POC tests performed on MSU were significantly more in agreement with the reference than when performed on FVU when analysis was done immediately. The error rate was 16% for MSU vs. 23% for FVU with POC culture, 27% vs. 40% with microscopy and 25% vs. 33% with dipstick testing. Delay of analysis up to four hours did not decrease agreement with the reference. Conclusion/Implication: MSU samples should be used in general practice for optimal accuracy of POC tests. Analysis can be delayed up to four hours. Key points Point-of-care tests (dipstick testing, microscopy and culture) for diagnosing urinary tract infection performed on mid-stream urine samples are significantly more accurate than when performed on first-void urine samples. Delay of analysis up to four hours did not decrease the accuracy of any of the point-of-care tests. Point-of-care culture was more accurate than dipstick and microscopy both when performed on mid-stream urine and first-void urine The main contaminant in first-void urine samples was Enterococci spp., which contributed to the majority of false positives.

[Investigation on endometrium from menstrual blood as a source of non-invasive tissue].

Objective: To investigate the feasibility of a non-invasive sampling method by collecting menstrual blood and obtaining endometrium for further research in menstruation-related diseases. Methods: On the second day of menstruation, menstrual blood was collected with menstrual cups for 4 hours, and the menstrual endometrium was filtered through a metal screen for weighing, cryopreserved, immunohistochemical staining and cell culture. Results: The collection process was painless and non-invasive. In the control group, the menstrual volume was (9.1±0.7) ml, and the endometrial tissue weight was (91.0±14.7) g. In the endometriosis group, the menstrual volume was (9.6±1.9) ml (P=0.022), and the endometrial tissue weight was (134.7±43.9) g (P=0.057). Endometrial cell culture was successful in all patients and should not be contaminated. The growth curve was a finite cell line type. The expression of cytokeratin 19 and vimentin in menstrual endometrium and cells were positive. Conclusions: By collecting menstrual blood and filtering endometrial tissue, it is an ideal non-invasive sampling method. In combination with advanced experimental technology, menstrual endometrium make further researches of endometriosis, endometrial lesions or other menstruation-related diseases possible.

[Survey of the acceptance status of HPV self-sampling screening in female population for cervical cancer].

Objective: To investigate the acceptance of HPV self-sampling mode in cervical cancer screening population and explore its feasibility. Methods: From March 5 to 15, 2018, researchers investigated women who participated in cervical cancer screening organized by Beijing Shunyi Women's and Children's Hospital in the form of questionnaires. Questionnaires were conducted on their acceptance status and the factors that affect the self-sampling experience. The specific contents of the questionnaires were as follows: (1) the experience of using self-sampling included operability, comfortable, sample time-consuming, bleeding or not after sampling; (2) psychological changes after self-sampling, including the willingness to accept self-sampling again, the worrying problems during self-sampling process. According to whether or not have operating video guidance, the self-sampling experience and psychological changes after self-sampling were compared. Results: (1) There were 1 375 women participated in the questionnaire survey, and 86.55% (1 190/1 375) of them thought the self-sampling was convenient, 78.40% (1 078/1 375) thought it was not uncomfortable, 88.58% (1 218/1 375) thought the sampling time was fast (less than 5 minutes), 94.04% (1 293/1 375) self-sampling without bleeding; and 83.27% (1 145/1 375) were willing to self-sampling for cervical cancer screening again, 85.82% (1 180/1 375) were afraid of inaccurate sampling. (2) Among the 1 375 women, 1 202 were in the video guidance group and 173 were in the non-guidance group. The self-sampling experience of women in video guidance group was better than those of non-guidance group in operability, comfortable, sampling time-consuming and bleeding after sampling. The proportion of women who willing to self-sampling again was higher than that of non-guidance group (86.69% vs 59.54%, respectively). The proportion of women who worried operating incorrectly was lower than that of non-guidance group (11.23% vs 32.37%, respectively). The differences were significant (all P<0.05). Conclusions: Self-sampling for HPV testing in cervical cancer screening is easy to operate and has little discomfort complaint. It is feasible in cervical cancer screening. Operational video guidance during the screening process could effectively improve the women's experience and willingness to self-sampling again in the future.

Differential effect of surgical manipulation on gene expression in normal breast tissue and breast tumor tissue.

BACKGROUND: Gene expression profiles of normal and tumor tissue reflect both differences in biological processes taking place in vivo and differences in response to stress during surgery and sample handling. The effect of cold (room temperature) ischemia in the time interval between surgical removal of the specimen and freezing is described in a few studies. However, not much is known about the effect of warm (body temperature) ischemia during surgery. METHODS: Three women with primary operable breast cancer underwent in situ biopsies from normal breast and tumor tissue prior to radical mastectomy. Ex vivo biopsies from normal and tumor tissue were collected immediately after surgical excision. The putative effects on gene expression of malignancy (tumor versus normal), surgical manipulation (post- versus pre-surgical) and interaction between the two (differences in effect of surgical manipulation on tumor and normal samples) were investigated simultaneously by Generalized Estimating Equation (GEE) analysis in this self-matched study. RESULTS: Gene set enrichment analysis (GSEA) demonstrates a marked difference in effect of surgical manipulation on tumor compared to normal tissue. Interestingly, a large proportion of pathways affected by ischemia especially in tumor tissue are pathways considered to be specifically up regulated in tumor tissue compared to normal. CONCLUSION: The results of this study suggest that a large contribution to this differential expression originates from altered response to stress in tumor cells rather than merely representing in vivo differences. It is important to bear this in mind when using gene-expression analysis to deduce biological function, and when collecting material for gene expression profiling.