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Being able to appropriately perform fine needle aspiration (FNA) collecting techniques and sample preparation is essential in obtaining a diagnostic sample, which is a critical skill for veterinary practitioners. Collection and preparation of cytologic samples are skills gained through practice. Experience leads to refinement of technique and improved diagnostic quality. Using live patients for mass skills training is not feasible; therefore, an aspiration simulation model and laboratory session was developed to reinforce physical exam skills, appropriate selection of sample collection supplies, and collection technique. Materials for the models include Ping-Pong balls, silicone, instant vanilla pudding mix, water, and stuffed animals. The laboratory session allows veterinary students to practice lesion identification, isolation, aspiration, and successful preparation. Subsequent submission of the collected sample involves being able to expel and spread the sample on a slide and proper labeling. While the simulation experience was initially developed for a short course with 12 students, it has recently been incorporated into the required clinical pathology clinical year rotation for up to 100 fourth-year veterinary students. The model is inexpensive and efficient and allows for technique development and immediate instructor assessment and feedback.

The sacrum is often used for sex estimation of human remains. However, reported sexual classification accuracies based on the sacrum produce highly discrepant estimates ranging from 54% to 98% depending on the method and reference sample. Here, we therefore aim to evaluate the sacrum's efficacy in differentiating males from females by comparing three different approaches in a homogeneous Central European sample (n = 58, 27 males and 31 females) obtained from the 19th century Weisbach collection. Specifically, we investigated the sacrum by 1) a qualitative visual inspection, 2) traditional linear metrics, namely, sacral width, corpus width, and the associated corporo-basal index, and 3) geometric morphometrics (GM) using a 3D configuration of 100 landmarks and semilandmarks. Classification accuracies for the qualitative approach ranged from 69% to 81%. The investigated quantitative methods based on linear dimensions led to comparable classification accuracies of 62% for sacral width and 78% for the corporo-basal index. However, absolute corpus width had a stronger discriminative power (86%), similar to the PC scores from shape space (87%). Unexpectedly, the GM approach, which considered the shape of the entire sacrum, did not classify better than linear variables. This was exceeded only when sacrum shape was combined with corpus width or another measure of size, like the natural logarithm of the centroid size, yielding a classification accuracy of 95%. The male and female group means differed mainly in the general height-to-width relationship and corpus-to-alae proportions, corroborating patterns previously described in the literature. Our results suggest that the sacrum is markedly less effective than the hipbone for sex determination, which is at odds with its central position within the pelvic girdle and thus its supposed obstetric relevance.

Background Multi-sample comparison is commonly used in cancer genomics studies. By using next-generation sequencing (NGS), a mutation's status in a specific sample can be measured by the number of reads supporting mutant or wildtype alleles. When no mutant reads are detected, it could represent either a true negative mutation status or a false negative due to an insufficient number of reads, so-called "coverage". To minimize the chance of false-negative, we should consider the mutation status as "unknown" instead of "negative" when the coverage is inadequately low. There is no established method for determining the coverage threshold between negative and unknown statuses. A common solution is to apply a universal minimum coverage (UMC). However, this method relies on an arbitrarily chosen threshold, and it does not take into account the mutations' relative abundances, which can vary dramatically by the type of mutations. The result could be misclassification between negative and unknown statuses. Methods We propose an adaptive mutation-specific negative (MSN) method to improve the discrimination between negative and unknown mutation statuses. For a specific mutation, a non-positive sample is compared with every known positive sample to test the null hypothesis that they may contain the same frequency of mutant reads. The non-positive sample can only be claimed as “negative” when this null hypothesis is rejected with all known positive samples; otherwise, the status would be “unknown”. Results We first compared the performance of MSN and UMC methods in a simulated dataset containing varying tumor cell fractions. Only the MSN methods appropriately assigned negative statuses for samples with both high- and low-tumor cell fractions. When evaluated on a real dual-platform single-cell sequencing dataset, the MSN method not only provided more accurate assessments of negative statuses but also yielded three times more available data after excluding the “unknown” statuses, compared with the UMC method. Conclusions We developed a new adaptive method for distinguishing unknown from negative statuses in multi-sample comparison NGS data. The method can provide more accurate negative statuses than the conventional UMC method and generate a remarkably higher amount of available data by reducing unnecessary “unknown” calls.

Forensic sex estimation is a key component in attempts to identify human remains. The skull is often used for this purpose, but is often damaged or recovered in fragments. Studies of individual cranial bones are hampered by subjectivity in visual assessment and difficulties in the ability to capture complex shape variations, as well as accounting for population-specificity in the expression of sexual dimorphism. The present study employed geometric morphometric analysis to assess morphological variation of the occipital bones of 792 adult South African individuals. Though sexual differences in size were detected, shape dimorphism was not detected until ancestry was simultaneously accounted for. Female occipital bones were less robust but more sloped than those of males. Detected variation between ancestry groups was sufficient to produce approximately 80% classification accuracies for Black and White groups, and 74% for the highly heterogeneous Coloured group. When variation was assessed according to sex and ancestry simultaneously, groups could be distinguished with 74.8% accuracy. Shape variation showed no significant age or secular trend changes, though some weak positive allometric influence was detected in the variation between sexes and between the Coloured group and the other two ancestry groups. This study demonstrated that even isolated bones such as the occipital could be used for sex and ancestry estimation, though population differences must be considered. The geometric morphometric analysis was found to allow objective and sufficiently sensitive detection of variation in the complex occipital shape to allow for accurate distinction of even highly heterogeneous and closely related groups, even when using a limited number of cranial landmarks. Thus, similar analyses of the occipital may be employed in forensic cases where the cranium has been damaged or only the occipital bone is recovered and available for analysis.

INTRODUCTION We aimed to evaluate the performance of the fully automated multiparameter CN-6000 hemostasis analyzer. METHODS Performance evaluation of the CN-6000 analyzer was conducted for 10 tests including prothrombin time (PT), activated partial prothrombin time (aPTT), fibrinogen level, anti-Xa activity, and antithrombin activity using a unique portfolio of liquid ready-to-use reagents. Precision, sample and reagent carryovers, throughput, and sample turnaround time (STAT) function were prospectively assessed. Results from 343 samples (normal subjects, critically ill patients, patients receiving anticoagulants, subjects with high or low fibrinogen levels, and patients with decreased levels of factor II, V, VII, and X) were compared to those obtained on the STA-R Max 2® analyzer using dedicated reagents. RESULTS Total precision (coefficient of variation) was below 7% for all parameters in both normal and pathological ranges. For all analyzed parameters, results obtained on the CN-6000 were strongly correlated with those obtained on the STA-R Max 2®analyzer. Agreement between both instruments was excellent for all assays. The CN-6000 demonstrated a 30% higher throughput compared to the STA-R Max 2® (258 vs 185 tests per hour for a panel of tests including PT, aPTT, fibrinogen, factor V, anti-Xa, and D-Dimer). STAT turnaround time for critical care samples testing was

One of the most challenging aspects of writing multiple-choice test questions is identifying plausible incorrect response options—i.e., distractors. To help with this task, a procedure is introduced that can mine existing item banks for potential distractors by considering the similarities between a new item’s stem and answer and the stems and response options for items in the bank. This approach uses natural language processing to measure similarity and requires a substantial pool of items for constructing the generating model. The procedure is demonstrated with data from the United States Medical Licensing Examination (USMLE®). For about half the items in the study, at least one of the top three system-produced candidates matched a human-produced distractor exactly; and for about one quarter of the items, two of the top three candidates matched human-produced distractors. A study was conducted in which a sample of system-produced candidates were shown to 10 experienced item writers. Overall, participants thought about 81% of the candidates were on topic and 56% would help human item writers with the task of writing distractors.

Background Artificial intelligence can assist in cardiac image interpretation. Here, we achieved a substantial reduction in time required to read a cardiovascular magnetic resonance (CMR) study to estimate left atrial volume without compromising accuracy or reliability. Rather than deploying a fully automatic black-box, we propose to incorporate the automated LA volumetry into a human-centric interactive image-analysis process. Methods and results Atri-U, an automated data analysis pipeline for long-axis cardiac cine images, computes the atrial volume by: (i) detecting the end-systolic frame, (ii) outlining the endocardial borders of the LA, (iii) localizing the mitral annular hinge points and constructing the longitudinal atrial diameters, equivalent to the usual workup done by clinicians. In every step human interaction is possible, such that the results provided by the algorithm can be accepted, corrected, or re-done from scratch. Atri-U was trained and evaluated retrospectively on a sample of 300 patients and then applied to a consecutive clinical sample of 150 patients with various heart conditions. The agreement of the indexed LA volume between Atri-U and two experts was similar to the inter-rater agreement between clinicians (average overestimation of 0.8 mL/m2 with upper and lower limits of agreement of − 7.5 and 5.8 mL/m2, respectively). An expert cardiologist blinded to the origin of the annotations rated the outputs produced by Atri-U as acceptable in 97% of cases for step (i), 94% for step (ii) and 95% for step (iii), which was slightly lower than the acceptance rate of the outputs produced by a human expert radiologist in the same cases (92%, 100% and 100%, respectively). The assistance of Atri-U lead to an expected reduction in reading time of 66%—from 105 to 34 s, in our in-house clinical setting. Conclusions Our proposal enables automated calculation of the maximum LA volume approaching human accuracy and precision. The optional user interaction is possible at each processing step. As such, the assisted process sped up the routine CMR workflow by providing accurate, precise, and validated measurement results.

All banknotes have security features which are intended to help determine whether they are false or genuine. Typically, however, the general public has limited knowledge of where on a banknote these security features can be found. Here, we tested whether counterfeit detection can be improved with the help of salient elements, designed to guide bottom-up visuospatial attention. We also tested the influence of the participant’s a priori level of trust in the authenticity of the banknote. In an online study (N = 422), a demographically diverse panel of Dutch participants distinguished genuine banknotes from banknotes with one (left- or right-sided) counterfeited security feature. Either normal banknotes (without novel design elements) or banknotes that contained a salient element (a pink rectangular frame) were presented for 1 s. To manipulate the participant’s level of trust, trials were administered in three blocks, whereby at the start of each block, participants were instructed that either one third, one half, or two thirds of the upcoming banknotes were counterfeit (though the true ratio was always 1:1). We hypothesized (i) that in the presence of a salient element, counterfeits would be better detected when the location of the salient element aligned with the location of the counterfeited security feature—i.e. that it would act as an attentional cue; and (ii) that this effect would be stronger with lower trust. Our hypotheses were partly confirmed: counterfeit detection improved with ‘valid cues’ and decreasing trust, but the level of trust did not modulate the cueing effect. As the overall detection performance was rather poor, we replicated the study with a sample of university students (N = 66), this time presenting stimuli until response. While indeed observing better overall performance, all other patterns were replicated. Our results provide evidence that attention can be guided to enhance banknote authentication.

Water consumed is stored in several water bodies in and around us, out of which dams accommodate a major portion of water. The quantity and quality monitoring of water in dams is troublesome due to their large surface area and high depths. Although groundwater resources are the primary water source in India, dams plays a vital role in water distribution and storage network. The Central Water Commission in India has identified more than 5,000 dams of which a major portion is persistently consumed by the rural and urban populations for drinking and irrigation. The water quality of these reservoirs is of serious concern as it would not only affect the socio-economic status of the nation but the aquatic systems as well. Water quality control and management are vital for a delivering clean water supply to the general society. Because of their size, collecting, assessing, and managing a vast volume of water quality data are critical. Water quality data are primarily obtained through manual field sampling; however, real-time sensor monitoring is increasingly being used for more efficient data collection. The literature depicts that the methods involving remote sensing and image processing of water quality analysis consume time, and require sample collection at various depths, analysis of collected samples, and manual interpretations. The objective of this study was to propose a novel cost-effective method to monitor water quality devoid of considerable human intervention. Sensor-based online monitoring aids in assessing the sample with limited technology, at various depths of water in the dam to analyze turbidity which gives the major indication of pure water. The quality analysis of the dam water is suitable if the water is assessed at the distribution end before consumption. Hence, to enhance the water management system, other quality parameters like pH, conductivity, temperature are sensed and monitored in the distribution pipeline. An unstable pH can alter the chemical and microbiological aspects of water, resulting in a variation of other water quality parameters. Temperature variations affect the amount of dissolved oxygen in the water bodies and results in unstable quality parameters. The change in dissolved solvents and the ionic concentration alters the electrical conductivity of the water and the increased concentration of salts also results in turbidity. The data from all the sensors are processed by the microcontroller, transmitted, and displayed in a mobile application comprehensible to the layman. HIGHLIGHTS The quantity and quality monitoring of water in dams is troublesome due to their large surface area and high depths.; An effective quality analysis is the demand of the hour and the problem statement is identified in developing a system to check water quality at various depths in a dam.; An efficient pipeline with various sensors which monitor turbidity, pH, and conductivity is developed as a prototype low cost device.

Rapid DNA platforms are fully automated systems capable of processing DNA from biological samples and interpreting the results in approximately 90 minutes with minimal human intervention. With a greater reliance on the system than on the analyst, validation data are especially needed to define the performance and limitations of commercially available Rapid DNA systems. Thus, validation studies of a Rapid DNA workflow consisting of the Applied Biosystems RapidHIT ID Instrument and RapidLINK software with a focus on the ACE GlobalFiler Express Sample Cartridge and reference buccal swabs were performed in accordance with Scientific Working Group on DNA Analysis Methods Validation Guidelines. These validation studies included assessments of sensitivity, contamination, concordance, reproducibility and repeatability, stability, inhibition, mixtures, sample reprocessing, precision, and first-pass success rate. Overall, the current Applied Biosystems RapidHIT ID Instrument with the ACE GlobalFiler Express sample cartridge was found to be a reliable tool for generation of STR profiles from reference-type buccal swabs. Supplementary Information The online version contains supplementary material available at 10.1007/s00414-021-02722-9.

Liquid-based preparation (LBP) cytology is commonly used in most laboratories these days due to its convenience and reliable results for the cervical cancer screening program. The PathTezt™ Liquid-based Pap smear is a second-generation LBP, which uses a filter-based concentration technique in processing the sample. Objective This study was done to evaluate the cellular fixation, morphology, quality of smear in gynae cytology, and diagnostic interpretation of cervical cytological smears produced by the PathTezt liquid-based processor. Materials and methods A total of 400 pap smear samples were taken and processed using the PathTezt 2000 processor. The slides were evaluated in terms of sample adequacy, percentage of the circle covered by epithelial cells, cellular distribution, obscuring factors, and cell fixation. Results About 95.25% (381) of the samples were satisfactory for the evaluation. In 19 (4.75%) of the samples, epithelial cells covered less than 50% of the circle. A sample with good cellular distribution was seen in 92% of the cases, while 354 (88.5%) samples showed minimal inflammatory background. Almost all the smears (95.75%) had no erythrocytes in the background. All smears showed good quality fixation features toward nuclear, cytoplasm, and microorganisms. The total performance rate was 99%. Conclusion Although the PathTezt liquid-based processor is still new compared to other first-generation LBP, the smears produced by this method were of high quality and it was cost-effective.

Polymerase chain reaction (PCR) testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is necessary for confirming a diagnosis of Coronavirus disease 2019 (COVID-19). Here we present a COVID-19 case of an elderly woman whose SARS-CoV-2 PCR tests showed false negative repeatedly by evaluating with different sampling sites and procedures. Nasopharyngeal swabs, suctioned sputum, and tongue swabs were collected for SARS-CoV-2-PCR. As for tongue swabs, we compared between two different sample conditions; one obtained with dry condition and the other obtained with moistened condition inside the oral cavity. SARS-CoV-2-PCR showed positive for an extended period with suctioned sputum samples compared with nasopharyngeal swabs and tongue swabs. No SARS-CoV-2 from a nasopharyngeal swab sample obtained on day 46 after symptoms onset was isolated despite high viral load (183740.5 copies/5μL). An adequate production of neutralizing antibody in a serum sample on day 46 was also confirmed. The number of RNA copies of the tongue swab samples was higher with moistened condition than with dry condition. The present case suggests that the difference of sampling site or sample condition can affect PCR results. High loads viral RNA detection does not always correlate with infectivity.

Background The Quality of Life Impact Refractive Correction (QIRC) questionnaire is a Rasch-validated instrument to assess the quality of life of ametropes with refractive correction. The original QIRC was validated in the United Kingdom. This study aimed to validate the Malay version of the QIRC among refractive correction wearers in Malaysia using Rasch analysis. Methods The original 20-item QIRC was forward-backward translated into Malay in preparation for the Pilot Malay QIRC. The pilot version was pre-tested on 105 spectacle/contact lens-corrected myopes, and the results were reviewed and cross-culturally adapted to produce the Final Malay QIRC. The final version was self-administered to a new sample of 304 participants. A Rasch analysis was conducted to evaluate the items and response categories of the Pilot and the Final Malay QIRC. Test-retest reliability was also analysed on the Final Malay QIRC. Results Based on the pre-test findings, Rasch analysis revealed a multidimensional scale (functional scale [Items 1 to 13] and emotional scale [Items 14 to 20], which were separated in subsequent analysis), unordered response categories for the functional scale (Category 3 was collapsed into Category 2), one misfit item (Item 3 was removed) and six items required modification (Items 4, 6 to 9, and 12 were reworded and cross-culturally adapted). In the Final Malay QIRC, both the functional and emotional scales had ordered response categories, good person reliability (functional, 0.80; emotional, 0.81) and separation index (functional, 2.01; emotional, 2.06), well-targeted items (targeting precision: functional, 0.28 logits; emotional, 0.08 logits), and satisfactory fit statistics (infit and outfit mean square were less than 1.50 for all items). A noticeable differential item functioning (DIF) between genders was found in Item 18 (DIF contrast, 0.40 logits; p = 0.04). Test-retest reliability analysis demonstrated a high intraclass correlation coefficient (0.94) and Cronbach’s alpha (0.97) with a coefficient of repeatability of ±8.14 units. Conclusions The Malay-translated version of the QIRC has good psychometric characteristics for assessing the quality of life of refractive correction wearers in Malaysia. This translated and cross-culturally adapted Malay QIRC is a valid and reliable instrument that can be used in routine clinical practice.

Background Non-standard body fluids (NSBFs) can provide essential clinical information otherwise unobtainable with conventional biological specimens. However, as most commercial chemistry reagents are only validated for serum, plasma, and urine by manufacturers, individual laboratories have to validate testing with NSBF to comply with regulatory standards. However, the heightened level of oversight and uncertainty of validation requirements to comply with regulatory standards pose a significant challenge for NSBF testing in clinical laboratories. Methods 28 combinations of high-volume chemistry tests requested on NSBF with established clinical utility were selected from retrospective data analysis. Specimens were analyzed with both closed and open channel chemistry reagents on a LABOSPECT 008AS platform (Hitachi High-Tech Co., Tokyo, Japan). Recovery studies were performed using a high concentration serum sample and 5 clinical NSBF samples at varying concentrations for each analyte. Acceptable performance limits were defined as 100 ± 10% of expected recovery. Results The average percent recovery ranged from 94.5% to 106.6% depending on the analyte/NSBF combination evaluated, and for each of the 28 combinations, the average percent recovery was within the predefined acceptable limits of ± 10%. Conclusions The recovery results from this study on the LABOSPECT 008AS platform demonstrates that any systematic matrix interference of high-volume chemistry testing on NSBF samples is well within the defined limits of acceptability. This work also demonstrates recovery studies performed by an individual laboratory are practial and it is feasible to demonstrate compliance with regulatory requirements for accuracy of chemistry testing on NSBF samples.

Inspired by Locard's exchange principle, which states "every contact leaves a trace", a trace residue sampling strategy has been developed for the analysis of discarded drug packaging samples (DPS), as part of an early warning system for illicit drug use at large public events including music/dance festivals. Using direct analysis in real time/mass spectrometry and tandem mass spectrometry, rapid and high-throughput identification and characterization of a wide range of illicit drugs and adulterant substances was achieved, including in complex polydrug mixtures and at low relative ion abundances. A total of 1362 DPS were analyzed either off-site using laboratory-based instrumentation or on-site and in close to real time using a transportable mass spectrometer housed within a mobile analytical laboratory, with each analysis requiring less than 1 min per sample. Of the DPS analyzed, 92.2% yielded positive results for at least one of 15 different drugs and/or adulterants, including cocaine, MDMA, and ketamine, as well as numerous novel psychoactive substances (NPS). Also, 52.6% of positive DPS were found to contain polydrug mixtures, and a total of 42 different drug and polydrug combinations were observed throughout the study. For analyses performed on-site, reports to key stakeholders including event organizers, first aid and medical personnel, and peer-based harm reduction workers could be provided in as little as 5 min after sample collection. Following risk assessment of the potential harms associated with their use, drug advisories or alerts were then disseminated to event staff and patrons and subsequently to the general public when substances with particularly toxic properties were identified.

Enzyme-linked immunosorbent assays (ELISA), as one of the most used immunoassays, have been conducted ubiquitously in hospitals, research laboratories, etc. However, the conventional ELISA procedure is usually laborious, occupies bulky instruments, consumes lengthy operation time, and relies considerably on the skills of technicians, and such limitations call for innovations to develop a fully automated ELISA platform. In this paper, we have presented a system incorporating a robotic-microfluidic interface (RoMI) and a modular hybrid microfluidic chip that embeds a highly sensitive nanofibrous membrane, referred to as Robotic ELISA, to achieve human-free sample-to-answer ELISA tests in a fully programmable and automated manner. It carries out multiple bioanalytical procedures to replace the manual steps involved in classic ELISA operations, including the pneumatically driven high-precision pipetting, efficient mixing and enrichment enabled by back-and-forth flows, washing, as well as integrated machine vision for colorimetric readout. The Robotic ELISA platform has achieved a low limit of detection (LOD) of 0.1 ng/mL in the detection of a low sample volume (15 μL) of chloramphenicol (CAP) within 20 min without human intervention, which is significantly faster than that of the conventional ELISA procedure. Benefiting from its modular design and automated operations, the Robotic ELISA platform has great potential to be deployed for a broad range of detections in various resource-limited settings or high-risk environments, where human involvement needs to be minimized, while the testing timeliness, consistency and sensitivity are all desired.

Most patients with memory disorders appear to forget at a relatively normal rate. Hence testing retention beyond the initial test session becomes unnecessary. However, it is now well established that a subsample of patients, most notably people with epilepsy, can show substantially increased forgetting rates even when acquisition rate is normal, raising the need for new tests focused on the assessment of long-term forgetting. Our study is part of the process of developing two such tests. Both focus on the need to test the same person several times and address the problem that each successive test may interfere with the memory of the event being tested. Depending on conditions, such effects can be substantial and may be either positive or negative. The Crimes and Four Doors Tests tackle this problem by presenting easily memorised episodes or scenes, from which a different sample of features is tested at each delay by telephone. We apply these two novel tests to assess rates of forgetting in groups of people with epilepsy and a matched control group. Both the visual and verbal tests showed clear evidence of accelerated forgetting in the epilepsy group supporting the potential value of the tests as convenient and sensitive measures of the rate of long-term forgetting.

Forensic anthropologists tasked with identification of skeletal remains often have to set up new strategies to overcome the limitations of conventional individualizing markers. A sound acquaintance with non-metric traits is essential for a reliable distinction between normal variations and pathological or traumatic conditions, yet the role of cranial variants in the identification process is still somehow ill-defined. One hundred crania (50 males and 50 females) of known sex and age were selected from the Collezione Antropologica LABANOF (a documented contemporary skeletal collection) and non-metric traits were scored as present or absent and by side. The frequencies of 13 traits were used to calculate the compound probabilities to find an individual with an exact combination of cranial features in the worldwide population. The probabilities of the majority of the individuals (53%) are within the 1 out of 10 million–1 out of 1 million interval. However, a fair number of subjects (25%) of the sample have the probabilities falling into the 1 out of 1 billion–1 out of 100 million interval, while the probabilities of a small portion of the sample (10%) are less than 1 out of 1 billion. This pilot study illustrates that some combinations of cranial variants are quite rare and may represent potential evidence to discern presumptive identifications, when an appropriate set of traits is selected and antemortem data are available for comparison. However, further research on larger and various samples is needed to confirm or discard the use of combinations of cranial non-metric traits as individualizing markers.

To cope up with the medical complications, scientists and physicians rely more on digitized historical evidence. It helps them to identify the disease and to develop new drugs and strategies. The authors have designed a model called B-Map. It can detect and segmenting any foreign object in the brain using fuzzy rules. The model can detect objects such as cancer and brain tumor. The proposed work aims at designing a classifier. The classifier would help to detect all possible foreign objects using one application. B-Map has been compared with benchmark algorithms such as K-means and ANN. It was found that the proposed model performs significantly better than the current techniques. Original patients' sample reports are taken from various medical laboratories. The figure numbers are retained as in the paper. The proposed model is able to find the edges and segment different types of foreign objects or one can say unexpected developments. Figure 12 shows the outer edges of a section of a brain MRI. The patient's MRI very clearly shows Hydrocephalus. The same is segmented and shown in Fig. 13. Figure 14 shows a segment of benign development and 15 shows a cancerous development which are again successfully segmented by the proposed model. The data on which testing is done is clinical data of the original patients. As the patient's details and data cannot be shared the author's cannot upload the data in the repository. As soon as the research completes, a benchmark dataset will be created and uploaded in public domain so that researchers can access it.

Severe internal trauma results in millions of hospitalizations each year, including thousands of deaths caused by subsequent multiple organ failure. The majority of these deaths occur within the first 24 h, and thus, rapid diagnosis of internal trauma severity is necessary for immediate treatment. For early organ damage identification, diagnosis in point-of-care settings is crucial for rapid triage and treatment. Recent reports suggest that circulating histones may serve as a biomarker for severe organ damage and the risk of multiple organ failure. Here, we report a point-of-care diagnostic system that utilizes the inherent interactions between histones and DNA for the fluorescence-based detection of histones in whole blood. In the assay, histones within the sample are wrapped by DNA, thus preventing an intercalating dye from binding the DNA and fluorescing. To allow for quantitative fluorescent measurements to be made in a point-of-care setting, we integrate a rapid, automated blood separation step into our assay. Furthermore, we eliminate manual reagent additions using a thermally responsive alkane partition (TRAP), thus making the system sample-to-answer. Finally, we demonstrate the assay in a portable fluorescence reader compatible with a point-of-care environment. We report a limit of detection 112 ng/mL in whole blood, suggesting that our device can be used to rapidly diagnose internal trauma severity and the likelihood of multiple organ failure in near-patient settings.

The Inhibitory Cascade Model was proposed by Kavanagh and colleagues (Nature, 449, 427-433 [2007]) after their experimental studies on the dental development of murine rodent species. These authors described an activator-inhibitor mechanism that has been employed to predict evolutionary size patterns of mammalian teeth, including hominins. In the present study, we measured the crown area of the three lower permanent molars (M1, M2, and M3) of a large recent modern human sample of male and female individuals from a collection preserved at the Institute of Anthropology of the University of Coimbra (Portugal). The main aim of the present study is to test if the size molar patterns observed in this human sample fits the Inhibitory Cascade Model. For this purpose, we first measured the crown area in those individuals preserving the complete molar series. Measurements were taken in photographs, using a planimeter and following well-tested techniques used in previous works. We then plot the M3 /M1 and M2 /M1 size ratios. Our results show that the premise of the Inhibitory Cascade Model, according to which the average of the crown area of M2 is approximately one-third of the sum of the crown area of the three molars, is fulfilled. However, our results also show that the individual values of a significant number of males and females are out of the 95% confidence interval predicted by the Inhibitory Cascade Model in rodents. As a result, the present analyses suggest that neither the sample of males, nor that of females, nor the pooled sample fits the Inhibitory Cascade Model. It is important to notice that, although this model has been successfully tested in a large number of current human populations, to the best of our knowledge this is the first study in which individual data have been obtained in a recent human population rather than using the average of the sample. Our results evince that, at the individual level, some factors not yet known could interfere with this model masking the modulation of the size on the molar series in modern humans. We suggest that the considerable delay in the onset of M3 formation in modern humans could be related to a weakening of the possible activation/inhibition process for this tooth. Finally, and in support of our conclusions, we have checked that the absolute and relative size of M1 and M2 is not related to the M3 agenesis in our sample. In line with other studies in primates, our results do not support the Inhibitory Cascade Model in a recent human sample. Further research is needed to better understand the genetic basis of this mechanism and its relationship to the phenotype. In this way, we may be able to find out which evolutionary changes may be responsible for the deviations observed in many species, including Homo sapiens.

Objectives: To evaluate four sample treatments in a safe and straightforward procedure to detect SARS-CoV-2 in saliva. Methods: Four sample treatments were evaluated in a 3-step procedure to detect SARS-CoV-2 in saliva: 1) heating at 95 °C for 5 min for sample inactivation; 2) sample treatment; 3) analysis by reverse-transcription loop-mediated isothermal amplification (LAMP). Saliva samples used were from infected individuals or were spiked with known quantities of viral particles. Results: Three treatments had a limit of detection (LOD) of 500.000 viral particles per ml of saliva and could be used to detect individuals with potential to transmit the disease. The treatment of phosphate buffer, dithiothreitol, ethylenediaminetetraacetic acid and proteinase K, with an additional 95 °C heating step, yielded a lower LOD of 95; its sensitivity ranged from 100% in patients with nasopharyngeal swab reverse-transcriptase polymerase chain reaction cycle threshold values 30. Conclusions: This report highlights the importance of an adequate sample treatment for saliva to detect SARS-CoV-2 and describes a flexible procedure that can be adapted to point-of-care. Although its sensitivity when LAMP is used is lower than reverse-transcriptase polymerase chain reaction, this procedure can contribute to COVID-19 control by detecting individuals able to transmit the disease.

DNA extraction is of utmost importance in archaeobiology, as it determines the success of further DNA analyses. This study concentrates on the success of ancient DNA extraction using silica spin columns and PCR-based analysis from archaeological skeletal material and investigates the influence of sample quantity, lysis time and lysis temperature during sample preparation. The results show that lysis times ranging from 2 to 48 h are suitable, and that lysis should be carried out at a constant temperature of 56°C. Concerning sample quantity, 10 mg for mitochondrial DNA and 50 mg for chromosomal DNA are sufficient for high quality analyses. Thus invaluable sample material can be saved, and time of sample preparation can be reduced considerably.

Background: Determine the exact slot dimension of a sample of a MBT prescription stainless steel conventional brackets from different manufacturers to compare the actual values with the nominal ones declared by the manufacturers and to verify the compliance with tolerance limits given by the ISO 27020:2019. Different batches from each manufacturer were evaluated to determine whether or not they are different in size. In addition, the geometry of the slot walls was assessed. Methods: 360 stainless steel preadjusted orthodontic brackets of 12 different manufacturers were assessed. All brackets had a nominal slot size of 0.022 by 0.028 inches, belonged to the right upper central incisor, and were fabricated with the metal injection molding technique (MIM). For each manufacturer, three different manufacturing batches were evaluated. Brackets were coded using a single-blind design. Results: All bracket systems in the study group except one displayed a statistically significant difference with the nominal declared value, although only four of the systems did not comply with the tolerance limits established by the ISO 27020:2019. In most of the systems, the slot height was oversized when compared to the nominal one. A significant interbatch variability was found in most of the evaluated systems. Most of the brackets walls were divergent. Conclusions: The dimensional accuracy of commercially available metal brackets is not guaranteed. The respect for the norm should be enforced as well as the quality controls along the manufacturing process since orthodontic brackets are a precision medical device. Objetivos: Determinar la dimensión exacta del slot de una muestra de brackets convencionales de acero inoxidable recetados MBT de diferentes fabricantes para comparar los valores reales con los nominales declarados por los fabricantes y verificar el cumplimiento de los límites de tolerancia dados por la norma ISO 27020:2019. Se evaluaron diferentes lotes de cada fabricante para determinar si eran o no de tamaño diferente. Además, se evaluó la geometría de las paredes de las ranuras. Métodos: Se evaluaron 360 brackets de ortodoncia preajustados de acero inoxidable de 12 fabricantes diferentes. Todos los brackets tenían un tamaño de ranura nominal de 0.022 por 0.028 pulgadas, pertenecían al incisivo central superior derecho y fueron fabricados con la técnica de moldeo por inyección de metal (MIM). Para cada fabricante se evaluaron tres lotes de fabricación diferentes. Los corchetes se codificaron utilizando un diseño simple ciego. Resultados: Todos los sistemas de brackets del grupo de estudio excepto uno presentaron una diferencia estadísticamente significativa con el valor nominal declarado, aunque solo cuatro de los sistemas no cumplieron con los límites de tolerancia establecidos por la norma ISO 27020:2019. En la mayoría de los sistemas, la altura de la ranura estaba sobredimensionada en comparación con la nominal. Se encontró una importante variabilidad entre lotes en la mayoría de los sistemas evaluados. La mayoría de las paredes de los soportes eran divergentes. Conclusiones: No se garantiza la precisión dimensional de los brackets metálicos disponibles en comercio. Se debe hacer cumplir el respeto a la norma y los controles de calidad a lo largo del proceso de fabricación ya que los brackets de ortodoncia son un dispositivo médico de precisión. Sin financiación 2.151 JCR (2020) Q3, 58/92, Dentistry, Oral Surgery & Medicine 0.620 SJR (2020) Q2, 47/141 Dentistry (miscellaneous) No data IDR (2020) UEV

Introduction: The evidence for using vacuum suction during EBUS is sparse and the optimal suction pressure for obtaining adequate samples has not yet been determined. Our aim was to assess the influence of suction on the adequacy and diagnostic yield of EBUS-TBNA. Material and methods: This single-center, prospective, randomized, non-inferiority trial assessed whether no-suction and 10 mL suction are inferior to 20 mL suction for adequacy and diagnostic yield of EBUS-TBNA aspirates. Results: Three hundred twenty three lymph nodes were sampled using EBUS-TBNA. Baseline characteristics of lymph nodes were comparable in the three suction groups. The overall adequacy of EBUS-TBNA aspirates in the no-suction, 10 mL, and 20 mL suction was 90%, 83.49%, and 77.88%, respectively. The differences in adequacy were 12.1% (95% CI: 3.9–20.3) and 5.6% (95% CI: –3.3–14.5) for no-suction vs. 20 mL, and 10 mL vs. 20 mL suction, respectively. No-suction and 10 mL were not inferior to 20 mL suction in terms of sample adequacy. At a superiority margin of 3.92%, no-suction was superior to 20 mL suction in terms of sample adequacy (p < 0.05). The overall diagnostic yield was comparable (63.6%, 52.3%, and 57.7% in 0, 10 mL, and 20 mL, respectively; p-value was not significant). The proportion of aspirates which were predominantly bloody was similar (no-suction—10.9%, 10 mL—13.8%, 20 mL—15.4%; p = 0.62). Conclusions: EBUS-TBNA with or without the application of vacuum suction does not influence specimen adequacy and diagnostic yield.

Introduction Urine collection from incontinent individuals can be challenging. Various methods have been devised to collect the sample without catheterization. Recently the PureWick external catheter was developed to draw the sample gently away from external female genitalia. While the primary purpose of the device is to prevent moisture and maintain skin integrity, the urine that is collected may be sent for laboratory analysis. We sought to validate the use of this collection method for common urine chemistry assays and urinalysis. Methods Twenty pools of residual urine samples were separated into “control” and “PureWick” treated samples. The control samples were maintained at room temperature while 15 mL of urine was added to the PureWick device which was connected to a vacuum line through a collection canister. The urine collected in the canister and the controls samples were all subject to urine chemistry strip, microscopic, and automated urine chemistry analysis. Results were compared between pairs of treated and control samples. Results No clear affect was noted on urine strip semi-quantitative or automated chemistry analysis from the PureWick collection. There was a statistically significant decrease in microscopic measurements of white blood cells and crystals in the PureWick urine samples. Discussion This study supports the use of the PureWick external catheter for collection of samples for most urinalysis and urine chemistry tests.

Background: Conventional home blood glucose measurements require a sample of blood that is obtained by puncturing the skin at the fingertip. To avoid the pain associated with this procedure, there is high demand for medical products that allow glucose monitoring without blood sampling. In this review article, all such products are presented. Methods: In order to identify such products, four different sources were used: (1) PubMed, (2) Google Patents, (3) Diabetes Technology Meeting Startup Showcase participants, and (4) experts in the field of glucose monitoring. The information obtained were filtered by using two inclusion criteria: (1) regulatory clearance, and/or (2) significant coverage in Google News starting in the year 2016, unless the article indicated that the product had been discontinued. The identified bloodless monitoring products were classified into three categories: (1) noninvasive optical, (2) noninvasive fluid sampling, and (3) minimally invasive devices. Results: In total, 28 noninvasive optical, 6 noninvasive fluid sampling, and 31 minimally invasive glucose monitoring products were identified. Subsequently, these products were characterized according to their regulatory, technological, and consumer features. Products with regulatory clearance are described in greater detail according to their advantages and disadvantages, and with design images. Conclusions: Based on favorable technological features, consumer features, and other advantages, several bloodless products are commercially available and promise to enhance diabetes management. Paths for future products are discussed with an emphasis on understanding existing barriers related to both technical and non-technical issues.

DNA structure has been leveraged in a variety of facets that allow scientists to perform a range of assays, including ones for identification of species, establishing evolutionary relationships between taxa, or even identifying individuals. Here, we present a DNA barcoding method as practical, hands-on approach that connects several experimental techniques in one sequence to teach the principles behind DNA isolation, purification, PCR, sequencing, and phylogeny analysis. Our set of exercises is designed for a teaching university laboratory setting. The three laboratory class assignments utilize DNA from a mushroom (can be purchased at a supermarket) and provide a pipeline to guide students through the process of identifying an unknown sample, like in many research laboratories. The third assignment can be used as a stand-alone exercise on phylogeny analysis and can be taught remotely. Students explore the theory behind the standard molecular techniques and apply it in a hands-on setting that involves experimental design, sample preparation, and use of hallmark molecular instruments.

OBJECTIVE According to the pristine conditions hypothesis, high-confidence identifications will be "remarkably accurate" when identification procedures (i.e., system variables, e.g., fair filler selection, double-blind administration, unbiased lineup instructions) are optimal, even if estimator variables (e.g., weapon presence, lighting, distance) are suboptimal (Wixted & Wells, 2017, p. 10). This has led some to conclude that estimator variables are not of much importance under those conditions. HYPOTHESIS We hypothesized that when multiple estimator variables are deficient, even high-confidence identifications will be less accurate than they would be when multiple estimator variables are optimal. METHOD With a sample of 2,191 college students (Mage = 20.14, 73% women), we conducted a strong test of this hypothesis by comparing a situation in which estimator variables were manipulated to produce either very good or very poor memory performance. RESULTS High-confidence suspect identifications were made significantly less frequently under poor viewing conditions than under good viewing conditions, and these differences are substantial if one assumes low base rates of guilt. CONCLUSIONS Estimator variables can be important for evaluating even high-confidence suspect identifications and establish some important boundary conditions for the pristine conditions hypothesis. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

Manual urine sediment analysis of a sample obtained from a 5 year old child by our clinical diagnostics laboratory revealed abundant “daisy-like” crystals, which have been first described in 2004 and found to be extremely rare in a follow-up publication by the same research group. To date only 12 samples have been described in the literature containing such crystals. Upon further investigation on how the sample was obtained, we were able to reproduce the process without any biological specimen involved. We show that these crystals are in fact contaminants from the sample collection recipient itself, which was a glass recipient sterilized by the patient’s family the night before sample collection, by boiling water with high calcium and magnesium content (hard water), and letting the recipient cool overnight with the water in it. The obtained abundant “daisy-like” crystals readily dissolve in acidic environment, and are composed most likely of mostly calcium carbonate. Sampling artifacts are therefore a possible explanation for at least some of the previously described “daisy-like” urinary crystals, as the formation of such crystals does not need to involve any biomolecules, only hard water and appropriate crystallization conditions for the limescale in it.

Objectives: In the cervical smear screening test as a sample collection method for liquid-based thin layer cytology, classically the collecting device is placed into a liquid fixative solution and vigorously swirled or rotated ten times in the solution and the collection device is removed from the solution. In this study, a plastic smear brush was used as the collection device. After the cervical cell sample was obtained, the smear brush was detached from the stick and left in the solution and given to the laboratory. Our aim in the study is to examine whether smear inadequacy rates have decreased with the method used in the study compared to the classical method. Material and methods: While the classical technique which the collecting device is placed into a solution and mixed and removed from the solution is defined as Method 1. The technique used in the study was defined as Method 2. The cervical smear screening test results obtained by two different methods in two consecutive time periods were analyzed. The two methods were compared using chi-square test in terms of smear inadequacy. Results: A total of 2129 test results, including 1129 smears in Method 1 and 1000 smears in Method 2 were examined. The mean ages of the patients tested in both methods were similar (36 ± 6.1 and 37 ± 6.7). Abnormal test result rate was similar for Method 1 and Method 2 (5.8% vs 4.9%, respectively). The inadequate sample rate was higher in Method 1 than Method 2 (8.3% vs 2.1%, respectively). Conclusions: The study showed that leaving the smear brush in the solution is a better way to reduce the inadequacy sample rates. This result may guide clinicians about smear techniques.

To perform repeated measurements in clinical practice without putting unnecessary strain on patients, short instruments are needed. The Brief Self-Report Measure of Agitation (BAM) is a short measure assessing agitation, which has been associated with suicidal ideation and behavior. However, its reliability and validity have not been examined in an independent sample. A non-clinical sample of N = 429 participants aged between 18-81 (

Background New developments in next-generation sequencing technologies and massive data received from this approach open wide prospects for personalised medicine and nutrition studies. Metagenomic analysis of the gut microbiota is paramount for the characterization of human health and wellbeing. Despite the intensive research, there is a huge gap and inconsistency between different studies due to the non-standardised and biased pipeline. Methodical and systemic understanding of every stage in the process is necessary to overcome all bottlenecks and grey zones of gut microbiota studies, where all details and interactions between processes are important. Results Here we show that an inexpensive, but reliable iSeq 100 platform is an excellent tool to perform the analysis of the human gut microbiota by amplicon sequencing of the 16 S rRNA gene. Two commercial DNA extraction kits and different starting materials performed similarly regarding the taxonomic distribution of identified bacteria. DNA/RNA Shield reagent proved to be a reliable solution for stool samples collection, preservation, and storage, as the storage of faecal material in DNA/RNA Shield for three weeks at different temperatures and thawing cycles had a low impact on the bacterial distribution. Conclusions Altogether, a thoroughly elaborated pipeline with close attention to details ensures high reproducibility with significant biological but not technical variations.

INTRODUCTION To perform comparability of instruments, the laboratory can select patient samples spanning the reportable range or can use plasma sets from commercial suppliers. We evaluated ExpertCor Routine (ECR) plasma set (Stago), a set of frozen plasmas enabling to verify the agreement between different coagulation analysers. Additionally, we evaluated whether the concept of transference of the reference range is acceptable between instruments, once comparability between the instruments is approved. METHODS Patient samples and the ECR plasma set were evaluated for method comparison for prothrombin time, activated partial thromboplastin time and fibrinogen on five instruments. Results of one instrument were compared to the mean of all analysers by Passing-Bablok regression and Bland-Altman analysis. Reference ranges were checked on all instruments. RESULTS The %mean difference was ≤5% and ≤3.7% for all analyser/parameter combinations, for ECR and patient sample data sets, respectively. All predefined criteria to fulfil good comparability between instruments were met. The between-instrument comparison with the ECR plasma set and the patient samples was equal for PT, INR and fibrinogen. After demonstrating comparability between instruments by either of the two plasma sample sets, reference ranges can be used interchangeably between identical instruments. CONCLUSION Instrument-to-instrument reproducibility showed comparable results using a data set obtained with patient samples or a commercial plasma set. Once comparability between instruments is confirmed, defined reference ranges can be transferred from one instrument to the other instrument without additional testing. The ECR plasma set is a good alternative to the use of local patient samples to evaluate instrument comparability.

We sampled nasal–pharyngeal throat swabs from 96,123 asymptomatic individuals at risk of SARS-CoV-2 infection, and generated 22,290 pools at collection, each containing samples from two to seven individuals. We detected SARS-CoV-2 in 24 pools, and confirmed the infection in 32 individuals after resampling and testing of 104 samples from positive pools. We completed the testing within 14 days. We would have required 64 days to complete the screening for the same number of individuals if we had based our testing strategy on individual testing. There was no difference in cycle threshold (Ct) values of pooled and individual samples. Thus, compared with individual sample testing, our approach did not compromise PCR sensitivity, but saved 77% of the resources. The present strategy might be applicable in settings, where there are shortages of reagents and the disease prevalence is low, but the demand for testing is high.

We examined the extent to which the Big Five domains, 30 facets, and nuances (uniquely represented by individual questionnaire items) capture age differences in personality, expecting domains to contain the least and nuances the most age-related information. We used an Internet sample (N = 24,000), evenly distributed between ages of 18 and 50 years and tested with a 300-item questionnaire. Separately based on domains, facets, and items, we trained models to predict age in one part of the sample and tested their predictive accuracy in another part. Big Five domains predicted age with an accuracy of r = .28, whereas facets' (r = .44) and items' (r = .65) predictions were more accurate. Less than 15% of the sample was needed to train models to their optimal accuracy. Residualizing the 300 items for all facets had no impact on their predictive accuracy, suggesting that age differences in specific behaviors, thoughts, and feelings (i.e., items) were not because of domains and facets but mostly unique to nuances. These findings replicated in a multisample dataset tested with another questionnaire. We found little evidence that age differences only appeared nuanced because items referred to age-graded roles or experiences. Therefore, a substantial part of personality development may be uniquely ascribed to narrow personality characteristics, suggesting the possibility for a many-dimensional representation of personality development. Besides theoretical implications, we provide concrete illustrations of how this can open new research avenues by enabling to study systematic variations between traits. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

Hemolysis, a common occurrence in blood collected for chemical analysis, has been reported to affect analytical test results for some analytes depending upon the material tested and the analytical technique employed. The potential for hemolysis to impact blood ethanol determinations using headspace gas chromatography of samples diluted with an internal standard was investigated. A sample of non-hemolyzed blood and a matched sample of hemolyzed blood were both analyzed thirty times for ethanol concentration using headspace gas chromatography. The mean ethanol concentration measured for the non-hemolyzed samples was 0.0639 g/dl. The mean ethanol concentration measured for the hemolyzed samples was 0.0642 g/dl. The calculated t value, 1.897, was less than the critical t value, 2.002, at a 0.05 level of significance. There was no measured statistical difference detected between the mean blood ethanol concentration determined for a hemolyzed whole blood sample and a non-hemolyzed whole blood sample.

Evaluating the short tandem repeat (STR) in the field is important for the timely identification of a suspect. Several lines showed that the RapidHIT® ID system is reliable for DNA genotyping with buccal swabs and naked DNA. However, the application of this approach with blood samples has been poorly investigated. Because blood samples are among the most common forensic samples in our laboratory, further studies should be conducted. Here, we assessed the analytical performance of 19 STR loci with a newly developed RapidINTEL (RI) Sample Cartridge Kit by using the blood samples with known genotypes. Several commonly used substrates were included in the sensitivity study, and FTA cards proved to be the most promising sample carrier for blood storage and later identification. There was superior sensitivity and specificity with a 100% concordance rate for 0.5 μL of blood or 7 ng of genomic DNA. The performance for blood samples was comparable with that for the standard protocol. High success rate (90.57%) and high-concordance (100%) genotyping were automatically achieved over a wide range of operating conditions except for TH01. No contamination was observed throughout the study. Hematin, indigo, and humic acid had limited influence on the instrument system, while urea and melanin dramatically affected the genotyping results. Generally, the newly developed RI sample cartridge provided an alternative method for the STR genotyping of single-source blood samples over a wide range of operating conditions.

Efforts to contain the spread of SARS-CoV-2 have spurred the need for reliable, rapid, and cost-effective diagnostic methods which can be applied to large numbers of people. However, current standard protocols for the detection of viral nucleic acids while sensitive, require a high level of automation and sophisticated laboratory equipment to achieve throughputs that allow whole communities to be tested on a regular basis. Here we present Cap-iLAMP (capture and improved loop-mediated isothermal amplification) which combines a hybridization capture-based RNA extraction of gargle lavage samples with an improved colorimetric RT-LAMP assay and smartphone-based color scoring. Cap-iLAMP is compatible with point-of-care testing and enables the detection of SARS-CoV-2 positive samples in less than one hour. In contrast to direct addition of the sample to improved LAMP (iLAMP), Cap-iLAMP prevents false positives and allows single positive samples to be detected in pools of 25 negative samples, reducing the reagent cost per test to ~1 Euro per individual., Current SARS-CoV-2 diagnostic methods are sensitive yet poorly suited to testing whole communities on a regular basis. Here the authors present Cap-iLAMP that tests gargle lavage samples with an improved colorimetric RT-LAMP.

The assessment of tobacco withdrawal is important for both research and clinical purposes. This study describes the psychometric development of a revised version of the 28-item Wisconsin Smoking Withdrawal Scale (WSWS; Welsch et al., Experimental and Clinical Psychopharmacology, 1999, 7, p. 354). Because the different contexts of use sometimes permit only brief assessment, this revision has produced both a brief and longer form using an updated pool of candidate items. For the revised Wisconsin Smoking Withdrawal Scale 2 (WSWS2), a candidate pool of 37 items was developed to measure nine putative withdrawal constructs. The stem and wording of items were revised as was the response scale. Data for psychometric analyses were derived from three smoking cessation randomized clinical trials conducted at the University of Wisconsin Center for Tobacco Research and Intervention. Dimensionality, internal consistency, and item characteristic analyses of the candidate items were conducted in a derivation sample to ascertain the factor structure and to identify items that could be used in the WSWS2 scales. Confirmatory factor analyses (CFAs) of reduced item sets and factor structure were conducted in two validation samples along with reliability and validity analyses. Derivation and validation sample analyses yielded a longer version of the WSWS2 (WSWS2-L) with 19 items and six subscales (Craving, Negative Affect, Hunger, Sleep, Restlessness, and Concentration) and a brief 6-item version (WSWS2-B). In validation sample analyses, both the WSWS2-L and the WSWS2-B demonstrated good reliability and validity as well as good fit in CFAs. The WSWS2-L and WSWS2-B possess improved construct coverage, fewer items, and other enhancements relative to the WSWS. (PsycInfo Database Record (c) 2021 APA, all rights reserved).

Most quantitative research methods are based on measuring either the total or the free concentration of an analyte in a sample. However, this is often insufficient for the study of complex biological systems. The main objective of this research was to develop new methods for providing more information from samples: the free concentration (C(f)), the total concentration (C(t)), and the plasma binding capacity (PBC). Samples were processed using microextraction and ultrafiltration. For each of these techniques, two quantification procedures were used: addition of isotopically labeled standard and repeated analysis of the same sample. The new methods were validated by analyzing clinical samples and samples with known concentrations. Methods based on addition of labeled compound were found to be the fastest, and most reproducible. For analysis of clinical samples, methods based on microextraction were more sensitive and more accurate than those based on ultrafiltration. For analysis of pooled plasma samples, the overall accuracy of all approaches to determine PBC, testosterone C(f), and testosterone C(t) was between 94–109%, 87–113%, and 94–122% respectively. The new approach goes beyond a simple concentration measurement, giving more information from clinical samples, with great potential for personalizing drug dosage and therapy to the needs of individual patients.

In-depth LC–MS-based proteomic profiling of limited biological and clinical samples, such as rare cells or tissue sections from laser capture microdissection or microneedle biopsies, has been problematic due, in large, to the inefficiency of sample preparation and attendant sample losses. To address this issue, we developed on-microsolid-phase extraction tip (OmSET)-based sample preparation for limited biological samples. OmSET is simple, efficient, reproducible, and scalable and is a widely accessible method for processing ~200 to 10,000 cells. The developed method benefits from minimal sample processing volumes (1–3 μL) and conducting all sample processing steps on-membrane within a single microreactor. We first assessed the feasibility of using micro-SPE tips for nanogram-level amounts of tryptic peptides, minimized the number of required sample handling steps, and reduced the hands-on time. We then evaluated the capability of OmSET for quantitative analysis of low numbers of human monocytes. Reliable and reproducible label-free quantitation results were obtained with excellent correlations between protein abundances and the amounts of starting material (R(2) = 0.93) and pairwise correlations between sample processing replicates (R(2) = 0.95) along with the identification of approximately 300, 1800, and 2000 protein groups from injected ~10, 100, and 500 cell equivalents, resulting from processing approximately 200, 2000, and 10,000 cells, respectively.

In forensic anthropology, sex estimation is a fundamental step in assessing individualbiological profiles when analyzing human skeletons. Yet, current methods are notreliable enough to allow an accurate sex identification of highly fragmented, burnt,or subadult remains. This paper aims to investigate sexual dimorphism of the bonylabyrinth on both identified subadult and adult individuals. The bony labyrinth is ofparticular interest for sex estimation since it is alleged to complete size and maturationpre-pubertally and is located inside the petrous part of the temporal bone whichprotects it from taphonomic processes. The study was performed on 93 CT scans ofidentified individuals from two Belgian osteological collections (19–20th century) andfrom current pediatric images (Erasme hospital, Brussels). Linear and angular measurementswere taken on 2D slices of right bony labyrinths. Intra- and interobservers errormeasurements were calculated. Statistical tests were used to unravel any morphologicalvariations between subadult and adult bony labyrinths and to highlight differencesbetween females and males, separately in subadults and adults. Linear discriminantfunctions were established by cross-validation and tested on an independent samplefrom Belgium. Some measurements were significantly different between subadultsand adults, and between females and males within both subadult and adult samples.Univariate functions achieved 72.7% in subadults and 68.4% in adults whereas multivariateequations increased accuracy respectively up to 84.9% and 78.4%. This studyentails promising results to design a sex estimation method suitable for fragmentedand/or subadult remains. Further metric approaches are needed to explore bony labyrinthsexual dimorphism., info:eu-repo/semantics/published

Background: The use of clozapine demands regular monitoring of clozapine plasma concentrations and of white blood cell parameters. The delay between sending blood samples for analysis and receiving the results hinders clinical care. Point-of-care testing (POCT) can provide drug assay results within a few minutes. Aim: This study aimed to investigate the utility of a novel point-of-care device that can measure clozapine concentrations using capillary blood samples collected via a finger stick. Method: During a five-week period starting in June 2019 eligible patients were asked to provide a finger-stick capillary sample in addition to their usual venous blood sample. Samples were analysed by the novel point-of-care device and by the standard laboratory method. Capillary blood samples were tested by the MyCare™ Insite POCT analyser, and a quantitative measurement of clozapine concentration was provided within six minutes. Results: A total of 309 patients agreed to measurements by the two methods. Analysis revealed clozapine concentrations in venous blood as determined by the laboratory method ranged from 20 to 1310 ng/mL and by POCT from 7 to 1425 ng/mL. There was a strong positive correlation ( R = 0.89) between the results from the venous and the capillary sample methods. The slope of the association between standard assay and MyCare™ Insite was 1.0 with an intercept of –21 ng/mL, indicating minimal bias. Conclusion: Clozapine concentrations can be accurately measured at the point of care using capillary blood samples collected via a finger stick. This approach may be more acceptable than venous sampling to patients and, with almost instant results available, more useful to clinicians.

Capillary blood sampled as dried blood spot (DBS) has shown substantial potential as test matrix in sports drug testing in various different settings, enabling the analysis of numerous different drugs and/or their respective metabolites. In addition to established beneficial aspects of DBS specimens in general (such as the minimally invasive and non-intrusive nature, and simplified sample transport), a yet unexplored advantage of DBS in the anti-doping context could be the opportunity of preserving a source of information complementary to routine doping controls performed in urine or venous blood. Whenever follow-up investigations are warranted or required, frequently collected and stored (but yet not analyzed) DBS samples could be target-tested for the compound(s) in question, in order to contribute to results management and decision-making processes.

In pathology protocols, a tissue block, such as one containing a mouse brain or a biopsy sample from a patient, can produce several hundred thin sections. Substantial time may be required to analyse all sections. In cases of uncertainty regarding which sections to focus on, noninvasive scout imaging of intact blocks can help in guiding the pathology procedure. The scouting step is ideally done in a time window of minutes without special sample preparation that may interfere with the pathology procedures. The challenge is to obtain some visibility of unstained tissue structures at sub-10 µm resolution. We explored a novel x-ray tomosynthesis method as a way to maximise contrast-to-noise ratio, a determinant of tissue visibility. It provided a z-stack of thousands of images at 7.3 μm resolution (10% contrast, half-period of 68.5 line pairs/mm), in scans of 5-15 minutes. When compared with micro-CT scans, the straight-line tomosynthesis scan did not need to rotate the sample, which allowed flat samples, such as paraffin blocks, to be kept as close as possible to the x-ray source. Thus, given the same hardware, scan time and resolution, this mode maximised the photon flux density through the sample, which helped in maximising the contrast-to-noise ratio. The tradeoff of tomosynthesis is incomplete 3D information. The microtomosynthesis scanner has scanned 110 unstained human and animal tissue samples as part of their respective pathology protocols. In all cases, the z-stack of images showed tissue structures that guided sectioning or provided correlative structural information. We describe six examples that presented different levels of visibility of soft tissue structures. Additionally, in a set of coronary artery samples from an HIV patient donor, microtomosynthesis made a new discovery of isolated focal calcification in the internal elastic lamina of coronary wall, which was the onset of medial calcific sclerosis in the arteries.A microscopy version of the imaging method for 3D luggage screening has been adapted to image unstained pathology samples. Pathology tests of tissue samples are used for clinical diagnosis and for biomedical research. The tissue samples are often embedded in paraffin blocks and sectioned into many thin slices, which are then stained with the appropriate agents for light microscopy. Since each tissue block can produce several hundred thin sections, much time and labour is required to analyse all sections. Noninvasive scout imaging of intact blocks can help in guiding the pathology procedure. The scouting step is ideally done in a time window of minutes without special sample preparation that may interfere with the pathology procedures. The challenge is to obtain some visibility of unstained tissue structures at sufficient resolution. X-ray imaging is a promising tool to meet the challenge since x-rays can penetrate thick samples that are opaque to visible light. With x-ray imaging, a determinant of tissue visibility is the flux density of photons that illuminate the sample. We explored a novel x-ray tomosynthesis method as a way to maximise this factor. It provided a stack of thousands of cross-sectional images at 7.3 μm resolution (half-period of 68.5 line pairs/mm) in scans of 5-15 minutes. When compared with micro-CT scans (a widely used laboratory technology), this method did not need to rotate the sample, which allowed flat samples such as paraffin blocks to be kept as close as possible to the x-ray source. Thus, given the same hardware, scan time and resolution, this method maximised the photon flux density through the sample, which helped in improving the visibility of unstained tissue under x-ray. The tradeoff of the method is incomplete 3D information. Over 100 unstained human and animal tissue samples have been scanned with this method as part of their respective pathology protocols. In all cases, the stack of cross-sectional images showed tissue structures that guided pathology analysis or provided correlative structural information. We describe six examples that presented different levels of tissue visibility. Additionally, in a set of coronary artery samples from an HIV patient donor, microtomosynthesis made a new discovery of isolated focal calcification in the internal elastic lamina of coronary wall, which was the onset of medial calcific sclerosis in the arteries.

Background: Human papillomavirus (HPV)-based screening is rapidly replacing cytology as the cervical screening modality of choice. In addition to being more sensitive than cytology, it can be done on self-collected vaginal or urine samples. This study will compare the high-risk HPV positivity rates and sensitivity of self-collected vaginal samples using four different collection devices and a urine sample. Methods: A total of 620 women referred for colposcopy were invited to provide an initial stream urine sample collected with the Colli-Pee device and take two vaginal self-samples, using either a dry flocked swab (DF) and a wet dacron swab (WD), or a HerSwab (HS) and Qvintip (QT) device. HPV testing was performed by the BD Onclarity HPV Assay. Results: A total of 600 vaginal sample pairs were suitable for analysis, and 505 were accompanied by a urine sample. Similar positivity rates and sensitivities for CIN2+ and CIN3+ were seen for DF, WD, and urine, but lower values were seen for QT and HS. No clear user preferences were seen between devices, but women found urine easiest to collect, and were more confident they had taken the sample correctly. The lowest confidence in collection was reported for HS. Conclusions: Urine, a DF swab, and WD swab all performed well and were well received by the women, whereas the Qvintip and HerSwab devices were less satisfactory. Impact: This is the first study to compare five self-sampling methods in the same women taken at the same time. It supports wider use of urine or vaginal self-sampling for cervical screening.

Background The US Food and Drug Administration (FDA) issued a guidance for bacterial risk control strategies for platelet components in September 2019 that includes strategies using secondary bacterial culture (SBC). While an SBC likely increases safety, the optimal timing of the SBC is unknown. Our aim was to develop a model to provide insight into the best time for SBC sampling. Study design and methods We developed a mathematical model based on the conditional probability of a bacterial contamination event. The model evaluates the impact of secondary culture sampling time over a range of bacterial contamination scenarios (lag and doubling times), with the primary outcome being the optimal secondary sampling time and the associated risk. Results Residual risk of exposure decreased with increasing inoculum size, later sampling times for primary culture, and using higher thresholds of exposure (in colony-forming units per milliliter). Given a level of exposure, the optimal sampling time for secondary culture depended on the timing of primary culture and on the expiration time. In general, the optimal sampling time for secondary culture was approximately halfway between the time of primary culture and the expiration time. Conclusion Our model supports that the FDA guidance is quite reasonable and that sampling earlier in the specified secondary culture windows may be most optimal for safety.

Urine analysis via a toilet-based device can enable continuous health monitoring, a transformation away from hospital-based care towards more proactive medicine. To enable reliable sample collection for a toilet-attached analyzer, here a novel sample collector is proposed. The applicability of the proposed sample collector is validated for long-term use. Geometric parameters of the 3D-printed sample collector are optimized. The collected and leftover volumes are quantified for a range of urination speeds and design parameters. For long-term cyclic use, the protein concentrations of samples are quantified and the effectiveness of washing the sample collector is assessed., NA