False-positive Cases Research Articles

Collaborative evaluation study on 18 candidate diseases for newborn screening in 1.77 million samples.

Analytical and therapeutic innovations led to a continuous but variable extension of newborn screening (NBS) programmes worldwide. Every extension requires a careful evaluation of feasibility, diagnostic (process) quality and possible health benefits to balance benefits and limitations. The aim of this study was to evaluate the suitability of 18 candidate diseases for inclusion in NBS programmes. Utilising tandem mass spectrometry as well as establishing specific diagnostic pathways with second-tier analyses, three German NBS centres designed and conducted an evaluation study for 18 candidate diseases, all of them inherited metabolic diseases. In total, 1 777 264 NBS samples were analysed. Overall, 441 positive NBS results were reported resulting in 68 confirmed diagnoses, 373 false-positive cases and an estimated cumulative prevalence of approximately 1 in 26 000 newborns. The positive predictive value ranged from 0.07 (carnitine transporter defect) to 0.67 (HMG-CoA lyase deficiency). Three individuals were missed and 14 individuals (21%) developed symptoms before the positive NBS results were reported. The majority of tested candidate diseases were found to be suitable for inclusion in NBS programmes, while multiple acyl-CoA dehydrogenase deficiency, isolated methylmalonic acidurias, propionic acidemia and malonyl-CoA decarboxylase deficiency showed some and carnitine transporter defect significant limitations. Evaluation studies are an important tool to assess the potential benefits and limitations of expanding NBS programmes to new diseases.

Serum anti-tissue transglutaminase IgA and prediction of duodenal villous atrophy in adults with suspected coeliac disease without IgA deficiency (Bi.A.CeD): a multicentre, prospective cohort study

Whether coeliac disease in adults can be diagnosed with serology alone remains controversial. We aimed to evaluate the accuracy of serum anti-tissue transglutaminase IgA (tTG-IgA) in the diagnosis of coeliac disease. In this multicentre, prospective cohort study, adult participants (aged ≥18 years) with suspected coeliac disease without IgA deficiency who were not on a gluten-free diet and who had a local serum tTG-IgA measurement, were enrolled from Feb 27, 2018, to Dec 24, 2020, by 14 tertiary referral centres (ten from Europe, two from Asia, one from Oceania, and one from South America) to undergo local endoscopic duodenal biopsy. Local serum tTG-IgA was measured with 14 different test brands and concentration expressed as a multiple of each test's upper limit of normal (ULN), and defined as positive when greater than 1 times the ULN. The main study outcome was the reliability of serum tests for the diagnosis of coeliac disease, as defined by duodenal villous atrophy (Marsh type 3 or Corazza-Villanacci grade B). Histology was evaluated by the local pathologist, with discordant cases (positive tTG-IgA without duodenal villous atrophy or negative tTG-IgA with duodenal villous atrophy) re-evaluated by a central pathologist. The reliability of serum tests for the prediction of duodenal villous atrophy was evaluated according to sensitivity, specificity, positive predictive value, negative predictive value, and the area under the receiver operating characteristic curve (AUC) for categorical and continuous data. We enrolled 436 participants with complete local data on serum tTG-IgA and duodenal histology (296 [68%] women and 140 [32%] men; mean age 40 years [SD 15]). Positive serum tTG-IgA was detected in 363 (83%) participants and negative serum tTG-IgA in 73 (17%). Of the 363 participants with positive serum tTG-IgA, 341 had positive histology (true positives) and 22 had negative histology (false positives) after local review. Of the 73 participants with negative serum tTG-IgA, seven had positive histology (false negatives) and 66 had negative histology (true negatives) after local review. The positive predictive value was 93·9% (95% CI 89·2-98·6), the negative predictive value was 90·4% (85·5-95·3), sensitivity was 98·0% (95·3-100·0), and specificity was 75·0% (66·6-83·4). After central re-evaluation of duodenal histology in 29 discordant cases, there were 348 true positive cases, 15 false positive cases, 66 true negative cases, and seven false negative cases, resulting in a positive predictive value of 95·9% (92·0-99·8), a negative predictive value of 90·4% (85·5-95·3), a sensitivity of 98·0% (95·3-100·0), and a specificity of 81·5% (73·9-89·1). Either using the local or central definition of duodenal histology, the positive predictive value of local serum tTG-IgA increased when the serological threshold was defined at increasing multiples of the ULN (p<0·0001). The AUC for serum tTG-IgA for the prediction of duodenal villous atrophy was 0·87 (95% CI 0·81-0·92) when applying the categorical definition of serum tTG-IgA (positive [>1 × ULN] vs negative [≤1 × ULN]), and 0·93 (0·89-0·96) when applying the numerical definition of serum tTG-IgA (multiples of the ULN). Additional endoscopic findings included peptic gastritis (nine patients), autoimmune atrophic gastritis (three), reflux oesophagitis (31), gastric or duodenal ulcer (three), and Barrett's oesophagus (one). In the 1-year follow-up, a midgut ileum lymphoma was diagnosed in a woman on a gluten-free diet. Our data showed that biopsy could be reasonably avoided in the diagnosis of coeliac disease in adults with reliable suspicion of coeliac disease and high serum tTG-IgA. None.

Histopathological Correlates of Lobar Microbleeds in False-Positive Cerebral Amyloid Angiopathy Cases.

A definite diagnosis of cerebral amyloid angiopathy (CAA), characterized by the accumulation of amyloid β in walls of cerebral small vessels, can only be obtained through pathological examination. A diagnosis of probable CAA during life relies on the presence of hemorrhagic markers, including lobar cerebral microbleeds (CMBs). The aim of this project was to study the histopathological correlates of lobar CMBs in false-positive CAA cases. In 3 patients who met criteria for probable CAA during life, but showed no CAA upon neuropathological examination, lobar CMBs were counted on ex vivo 3T magnetic resonance imaging (MRI) and on ex vivo 7T MRI. Areas with lobar CMBs were next sampled and cut into serial sections, on which the CMBs were then identified. Collectively, there were 25 lobar CMBs on in vivo MRI and 22 on ex vivo 3T MRI of the analyzed hemispheres. On ex vivo MRI, we targeted 12 CMBs for sampling, and definite histopathological correlates were retrieved for 9 of them, of which 7 were true CMBs. No CAA was found on any of the serial sections. The "culprit vessels" associated with the true CMBs instead showed moderate to severe arteriolosclerosis. Furthermore, CMBs in false-positive CAA cases tended to be located more often in the juxtacortical or subcortical white matter than in the cortical ribbon. These findings suggest that arteriolosclerosis can generate lobar CMBs and that more detailed investigations into the exact localization of CMBs with respect to the cortical ribbon could potentially aid the diagnosis of CAA during life. ANN NEUROL 2023;94:856-870.

ThyroidPrint®: clinical utility for indeterminate thyroid cytology.

Molecular testing contributes to improving the diagnosis of indeterminate thyroid nodules (ITN). ThyroidPrint® is a 10-gene classifier aimed to rule-out malignancy in ITN. Post-validation studies are necessary to determine the real-world clinical benefit of ThyroidPrint® in patients with ITN. A single center, prospective, non-interventional clinical utility study was performed, analyzing the impact of ThyroidPrint® in the physicians' clinical decisions for ITN. Demographics, nodule characteristics, benign call rates (BCR) and surgical outcomes were measured. Histopathological data was collected from surgical biopsies of resected nodules. Of 1272 fine needle aspirations (FNA), 109 (8.6%) were Bethesda III and 135 (10.6%) Bethesda IV. Molecular testing was performed in 155 of 244 ITN (63.5%), of which 104 were classified as benign (BCR of 67.1%). After a median follow-up of 15 months, 103 of 104 (99.0%) patients with a benign ThyroidPrint® remained under surveillance and one patient underwent surgery which was a follicular adenoma. Surgery was performed in all 51 patients with a suspicious ThyroidPrint® result and in 56 patients that did not undergo testing, with a rate of malignancy of 70.6% and 32.1%, respectively. A higher BCR was observed in FLUS (87%) compared to AUS (58%) (p-value <0.05). False-positive cases included four benign follicular nodules, six follicular and four oncotytic adenomas. Our results show that, physicians chose active surveillance instead diagnostic surgery in all patients with a benign ThyroidPrint® result, reducing the need for diagnostic surgery in 67% in patients with preoperative diagnosis of ITN.

Biochemical and molecular characteristics among infants with abnormal newborn screen for very-long-chain acyl-CoA dehydrogenase deficiency: A single center experience

ObjectiveTo define the biochemical and molecular characteristics and diagnostic outcomes of a large US cohort of VLCAD deficiency positive cases as detected by newborn screening (NBS) with MS:MS. This relatively common disorder of fatty acid oxidation is screened for in every state in America and often results in extensive testing of multiple samples to arrive at a diagnostic conclusion. Materials and methodsWe compared NBS dried blood spot (DBS) acylcarnitine profile (ACP) C14, C14:1, C14:2, C14:1/C12:1 ratio and plasma C14, C14:1, C14:2, C14:1/C12:1, C14:1/C16 and C14:1/C2 ratios among true positive and false positive cases. Results of VLCAD enzyme analysis, molecular testing and fibroblast fatty acid oxidation probe assay were analyzed. ResultsThe presence of compound heterozygous or homozygous pathogenic variants, along with elevations of C14, C14:1 and C14:1/C12:1 ratio, identified 19 VLCAD deficiency cases. All were asymptomatic at most recent follow-up visits. The C14:1/C12:1 ratio in NBS-DBS ACP and plasma acylcarnitine profiles at follow-up (follow-up plasma ACP), is the most useful marker to differentiate between true and false positive cases. Among all cases with molecular analysis data available, approximately 56.7% had a single pathogenic mutation. Lymphocyte enzyme analysis (n = 61) was uninformative in 23% of cases studied. ConclusionVLCAD deficiency NBS by MS:MS is highly effective at identifying asymptomatic affected infants. Our cohort showed that elevation of C14:1/C12:1, in both NBS DBS and plasma ACP, was informative in discriminating affected from unaffected individuals and contributes to improve the accuracy of confirmatory testing of infants with presumptive positive for VLCAD deficiency.

Assessment of the Value of Mean Platelet Volume, Des Gamma Carboxy Prothrombin and Alpha Feto Protein in Early Detection of HCC

Abstract Background Hepatocellular carcinoma (HCC) is the most common primary malignant tumor of the liver. AFP is the gold standard tumor marker for HCC, mean platelet volume (MPV) is a parameter obtained from complete blood count (CBC) by automated analyzers, shown to be increased in multiple malignancies and inflammatory conditions. This cross sectional comparative study was designed to evaluate the diagnostic usefulness of MPV as a diagnostic marker in HCC patients due to cirrhosis. Aim of the Work To determine the value of mean platelet volume, Des gamma Carboxy prothrombin and Alfa-feto protein in early detection of Hepatocelluler carcinoma Patients and Methods 105 patients were enrolled in this study, they were divided into 3 equal groups Group A: Hepatocelluler carcinoma patients (HCC) Group B: Liver cirrhosis patients Group C: Healthy Control group each is 35 patients. MPV, AFP, DCP were evaluated in all groups. Ultrasound and Triphasic CT. were also done to Group A&amp;B to confirm or exclude the diagnosis of HCC and in group C as they are candidates for LDLT donors. Results Our results showed that regarding the diagnostic performance of the three tested tumor markers (AFP, MPV and DCP) in differentiating HCC from CLD groups that AFP had area under curve (AUC) of 0.719, Standard error (SE) of 0.061, Confidence interval (CI) of 0.599-0.839 and P value of 0.002 at a cut point of ≥ 40.0, MPV had area under curve (AUC) of 0.801, Standard error (SE) of 0.063, Confidence interval (CI) of 0.678 − 0.924 and P value of &amp;lt; 0.001 at a cut point of ≥ 10.2, while DCP had area under curve (AUC) of 0.842, Standard error (SE) of 0.047, Confidence interval (CI) of 0.750 − 0.935 and P value of &amp;lt; 0.001 at a cut point of ≥ 69.3 proving that DCP had the highest significant diagnostic performance in differentiating HCC from CLD groups also DCP≥ 69.3 had highest sensitivity(of 97.1%) and NPV(of 96.3%). AFP≥ 40.0 had highest specificity (of 91.4%) and PPV (of 86.4%). Youden’s index was highest in DCP ≥69.3, followed by MPV ≥10.2 and lowest in AFP ≥40.0. MPV had moderate sensitivity and specificity in between AFP and DCP with values of 71.4% and 85.7% respectively with also moderate PPV of 83.3 and NPV of 75% Conclusion Measurement of MPV is non-invasive, cheap and quick, and may therefore serve as a predictor of HCC in patients with CLD. Low sensitivity and specificity, on the other hand, suggests that this may be an adjunctive parameter to some other markers like AFP. Further studies with larger samples are needed to determine the association of MPV with HCC. For a single test DCP had highest significant diagnostic performance in differentiating HCC from CLD groups. In detection of HCC testing AFP≥ 40.0 if positive considered positive. If negative, retest for DCP≥ 69.3 if positive considered positive, otherwise is negative. This although deceased specificity and Positive predictive value, it raised sensitivity and Negative Predictive Value to 100.0% when combining both AFP and DCP. The benefit of increasing the sensitivity is not to miss cases with HCC while the decreased specificity and PPV can be compensated by further imaging studies to eliminate the false positive cases.

The Use of Saliva Samples to Test for Congenital Cytomegalovirus Infection in Newborns: Examination of False-Positive Samples Associated with Donor Milk Use.

A universal screening research study was conducted in six hospitals to identify the clinical sensitivity of polymerase chain reaction (PCR) testing on newborn dried blood spots (DBSs) versus saliva specimens for the diagnosis of congenital cytomegalovirus (cCMV). CMV DNA positive results from DBSs or saliva were confirmed with urine testing. Findings of several false-positive (FP) saliva PCR results prompted an examination of a possible association with donor milk. Documentation of the frequency of positive saliva results, including both true-positive (TP) and FP status from clinical confirmation, occurred. The frequency of donor milk use was compared for TP and FP cases. Of 22,079 participants tested between 2016 and 2022, 96 had positive saliva results, 15 were determined to be FP, 79 TP, and 2 were excluded for incomplete clinical evaluation. Newborn donor milk use was identified for 18 (19.14%) of all the positive saliva screens. Among the 15 FPs, 11 (73.33%) consumed donor milk compared to 7 of the 79 TPs (8.8%) (OR 28.29, 95% CI 7.10-112.73, p < 0.001). While milk bank Holder pasteurization inactivates CMV infectivity, CMV DNA may still be detectable. Due to this possible association, screening programs that undertake testing saliva for CMV DNA may benefit from documenting donor milk use as a potential increased risk for FP results.

Retrospective analysis of cell-free fetal DNA prenatal testing of maternal peripheral blood

To assess the value of non-invasive prenatal testing (NIPT) for trisomy 21 (T21), trisomy 18 (T18), trisomy 13 (T13), sex chromosome aneuploidies, chromosomal microdeletions and microduplications using cell-free fetal DNA from peripheral blood samples of pregnant women. A total of 15 237 pregnant women who had undergone NIPT testing at the Maternity and Child Health Care Hospital of Zaozhuang from February 2015 to December 2021 were enrolled in this study. For those with a high risk by NIPT, amniotic fluid samples were collected for G-banding chromosomal karyotyping analysis and chromosomal microarray analysis to verify the consistency of NIPT with results of prenatal diagnosis. All of the women were followed up by telephone for pregnancy outcomes. Among the 15 237 pregnant women, 266 (1.75%) were detected with a high risk for fetal chromosomal abnormality were detected. Among these, 79 (29.7%) were at a high risk for T21, 26 (9.77%) were at a high risk for T18, 9 (3.38%) were at a high risk for T13, 74 (27.82%) were at a high risk for sex chromosome aneuploidies, 12 (4.51%) were at a high risk for other autosomal aneuploidies, and 66 (24.81%) were at a high risk for chromosomal microdeletions or microduplications. 217 women had accepted invasive prenatal diagnosis and respectively 50, 13, 1, 25, 1 and 18 were confirmed with T21, T18, T13, sex chromosome aneuploidies, autosomal aneuploidies and microdeletions/microduplications, and the positive predictive values were 75.76%, 68.42%, 11.11%, 40.32%, 10% and 35.29%, respectively. For 13 042 women (85.59%), the outcome of pregnancy were successfully followed up. During the follow-up, one false negative case of T21 was discovered. No false positive cases for T13 and T18 were found. NIPT has a sound performance for screening T13, T18 and T21, and is also valuable for screening other autosomal aneuploidies, sex chromosome aneuploidies and chromosomal microdeletions/microduplications.

Python API Misuse Mining and Classification Based on Hybrid Analysis and Attention Mechanism

APIs play a crucial role in contemporary software development, streamlining implementation and maintenance processes. However, improper API usage can result in significant issues such as unexpected outcomes, security vulnerabilities and system crashes. To detect API misuses, current methods primarily rely on comparing established API usage patterns with target points for automated detection, mainly based on pre-validated datasets. Nonetheless, there is a scarcity of publicly available datasets on API misuses and their corresponding fixes, which hinders data-driven research. Moreover, most existing techniques concentrate on statically typed languages, such as Java and C, with only a few addressing dynamic languages like Python effectively, due to difficulties in handling dynamic features. Therefore, it is essential to identify Python API misuses and their fixes automatically and promptly. In this paper, we introduce HatPAM, a Hybrid Analysis and Attention-based Python API-Misuse Miner, which (a) provides a method for automatically mining true-positive commits related to Python API-misuse fixes from GitHub and (b) presents the subsequent processing for classifying Python API misuses in true-positive cases. Particularly, HatPAM applies hybrid static analysis and introduces a structure-based attention mechanism to examine syntax, semantics and structural features in Python code context, and considers the consistency between code and developers’ natural intent to significantly reduce false-positive cases. Evaluation on six popular Python projects reveals that HatPAM outperforms various state-of-the-art baselines, achieving up to 92.2% Precision, 86.7% Recall and 89.3% F1-score, indicating its capability to identify and classify Python API-misuse commits.

Added value of an artificial intelligence algorithm in reducing the number of missed incidental acute pulmonary embolism in routine portal venous phase chest CT.

The purpose of this study was to evaluate the incremental value of artificial intelligence (AI) compared to the diagnostic accuracy of radiologists alone in detecting incidental acute pulmonary embolism (PE) on routine portal venous contrast-enhanced chest computed tomography (CT). CTs of 3089 consecutive patients referred to the radiology department for a routine contrast-enhanced chest CT between 27-5-2020 and 31-12-2020, were retrospectively analysed by a CE-certified and FDA-approved AI algorithm. The diagnostic performance of the AI was compared to the initial report. To determine the reference standard, discordant findings were independently evaluated by two readers. In case of disagreement, another experienced cardiothoracic radiologist with knowledge of the initial report and the AI output adjudicated. The prevalence of acute incidental PE in the reference standard was 2.2% (67 of 3089 patients). In 25 cases, AI detected initially unreported PE. This included three cases concerning central/lobar PE. Sensitivity of the AI algorithm was significantly higher than the outcome of the initial report (respectively 95.5% vs. 62.7%, p < 0.001), whereas specificity was very high for both (respectively 99.6% vs 99.9%, p = 0.012). The AI algorithm only showed a slightly higher amount of false-positive findings (11 vs. 2), resulting in a significantly lower PPV (85.3% vs. 95.5%, p = 0.047). The AI algorithm showed high diagnostic accuracy in diagnosing incidental PE, detecting an additional 25 cases of initially unreported PE, accounting for 37.3% of all positive cases. Radiologist support from AI algorithms in daily practice can prevent missed incidental acute PE on routine chest CT, without a high burden of false-positive cases. • Incidental pulmonary embolism is often missed by radiologists in non-diagnostic scans with suboptimal contrast opacification within the pulmonary trunk. • An artificial intelligence algorithm showed higher sensitivity detecting incidental pulmonary embolism on routine portal venous chest CT compared to the initial report. • Implementation of artificial intelligence support in routine daily practice will reduce the number of missed incidental pulmonary embolism.

Diagnostic value of heart-to-mediastinum ratio in 99mTc-pyrophospate SPECT/CT for transthyretin cardiac amyloidosis

BackgroundIn transthyretin cardiac amyloidosis (ATTR-CA), 99mTc-pyrophosphate myocardial scintigraphy (99mTc-PYP) is a diagnostic tool that utilizes visual and quantitative evaluation. However, false positive cases can occur because of tracer accumulation in the blood. We investigated the effectiveness of the heart-to-mediastinum (H/M) ratio of 99mTc-PYP in ATTR-CA diagnosis. MethodsWe retrospectively included 164 patients who underwent 99mTc-PYP single-photon emission computed tomography/computed tomography between March 2019 and January 2022. The diagnostic accuracy of ATTR-CA was examined by the heart-to-contralateral lung (H/CL) and H/M ratio calculated at 3 hours post-tracer administration. ResultsAfter the exclusion of patients who did not undergo endomyocardial biopsy, 30 patients (15 each with ATTR-CA and without ATTR-CA) were included. The receiver operating characteristic curve used to distinguish ATTR-CA from non-ATTR-CA patients revealed an area under the curve of 0.986 and 0.943, respectively. A H/M ratio of > 1.41 identified ATTR-CA patients with a sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of 100, 93.3, 93.3, and 100%, respectively. Conversely, an H/CL ratio of > 1.3 identified ATTR-CA patients with 100% sensitivity, 40.0% specificity, 62.5% PPV, and 100% NPV. ConclusionThe H/M ratio obtained at 3 hours post-injection has the potential to be a novel indicator for ATTR-CA.

Role of Contrast-Enhanced Spectral Mammography in the detection of breast cancer

Objective: to assess the benefits of using contrast-enhancer spectral mammography in patients with an established category of BI-RADS 4a, 4b according to the results of a standard examination (digital mammography and ultrasound). Materials and methods. The date of 425 patients with a suspicious category of BI-RADS in the period from 2018 to 2021 were analyzed. As part of the additional examination, all patients underwent contrast-enhanced spectral mammography follow by histological verification of the identified changes. The sensitivity and specificity of the methods were evaluated. Results. The final diagnosis of breast cancer was established in 247 (58.1%) patients out of 425 women. Multicenter and multifocal growth was diagnosed in 17 (6.8%) patients out of 247, and primary multiple synchronous cancer was detected in 6 (2.4%) patients out of 247. False-positive cases when assessing CESM mainly consisted of: typical and atypical ductal hyperplasia, fibroadenoma, intraductal papilloma. Conclusion. Contrast-enhancer spectral mammography is a highly sensitive and highly specific method in the detection of malignant tumors of the breast. The method also allows minimizing the number of unwanted interventions in suspicious BI-RADS categories.

Diagnostic role of FNA cytology in the evaluation of cervical lymph nodes in thyroid cancers: Combined evaluation of thyroglobulin in eluate from FNA cytology.

The presurgical evaluation of cervical lymph nodes (CLNs) in the management of thyroid malignant lesions is crucial for the extent of surgery or detection of metastases. In these last decades, fine-needle aspiration cytology (FNAC) has been shown to have a central role in the detection of nodal thyroid metastases. It is adopted for the possibility of confirming suspected metastases either in the presurgical phase or in the follow-up of patients after thyroidectomy. However, FNAC from CLNs can be challenging, especially in cystic lesions. In this regard, the combination of FNAC with thyroglobulin measurement in the eluate from FNAC (Tg-FNAC) seems to increase the sensitivity of FNAC in the detection of CLN metastases. The role of FNAC and Tg-FNAC was investigated in this series. One hundred fifty-three prospective cytological samples of CLNs were studied along with surgical follow-up in the period between 2020 and 2022. Immunocytochemistry (ICC) was performed on liquid-based cytology-stored material. One hundred fifty-nine enlarged CLNs included 19 central lymph nodes and 140 CLNs. Forty-two thyroidal CLN metastases and 117 reactive lymph nodes were found. Thirty-one CLN dissections were performed in patients with a previous diagnosis of thyroid carcinoma (mostly papillary thyroid carcinoma [PTC]), whereas 128 CLNs with a concomitant suspicious and/or malignant thyroid nodule were found. There was one false-positive case among all the malignant histologically confirmed cases, and two of 117 reactive CLNs (1.7%) had a diagnosis of metastatic PTC. Markedly high Tg-FNAC was found in all metastatic CLNs, including 11 cystic metastatic CLNs detected by Tg-FNAC with a negative FNAC. ICC (including Tg, CK-19, and LCA) recognized nine cases with low Tg-FNAC and scant suspicious thyrocytes. Tg-FNAC plus FNAC diagnosed 94.2% of malignancies. FNAC represents a valid method for the evaluation of CLNs, especially combined with ICC. Tg-FNAC is an additional method with a useful role in FNAC.

Rapid Screening of Urinary Tract Infection Using Microfluidic Inertial-Impedance Cytometry.

Urinary tract infection (UTI) diagnosis based on urine culture for bacteriuria analysis is time-consuming and often leads to wastage of hospital resources due to false-positive UTI cases. Direct cellular phenotyping (e.g., RBCs, neutrophils, epithelial cells) of urine samples remains a technical challenge as low cell concentrations, and urine characteristics (conductivities, pH, microbes) can affect the accuracy of cell measurements. In this work, we report a microfluidic inertial-impedance cytometry technique for label-free rapid (<5 min) neutrophil sorting and impedance profiling from urine directly. Based on size-based inertial focusing effects, neutrophils are isolated, concentrated, and resuspended in saline (buffer exchange) to improve consistency in impedance-based single-cell analysis. We first observed that both urine pH and the presence of bacteria can affect neutrophil high-frequency impedance measurements possibly due to changes in nucleus morphology as neutrophils undergo NETosis and phagocytosis, respectively. As a proof-of-concept for clinical testing, we report for the first time, rapid UTI testing based on multiparametric impedance profiling of putative neutrophils (electrical size, membrane properties, and distribution) in urine samples from non-UTI (n = 20) and UTI patients (n = 20). A significant increase in cell count was observed in UTI samples, and biophysical parameters were used to develop a UTI classifier with an area under the receiver operating characteristic curve of 0.84. Overall, the developed platform facilitates rapid culture-free urine screening which can be further developed to assess disease severity in UTI and other urologic diseases based on neutrophil electrical signatures.

Factors influencing creatine kinase-MM concentrations in newborns and implications for newborn screening for Duchenne muscular dystrophy

IntroductionNewborn screening for Duchenne muscular dystrophy can be performed via a first-tier creatine kinase-MM measurement followed by reflex testing to second-tier molecular analysis of the DMD gene. In order to establish appropriate cut-offs for the creatine kinase-MM screen, factors that influence creatine kinase-MM in newborns were investigated. Materials and methodsCreatine kinase-MM data from a consented pilot study in New York State were collected over a two-year period and combined with de-identified validation data and analyzed. Univariate analysis and multiple linear regression analysis were performed. ResultsThe analysis indicated that age of newborn at specimen collection, gestational age and birth weight were significant influencers of CK-MM levels in newborns. In addition, to a lesser extent, sex, race/ethnicity and seasonal temperature also affect CK-MM levels in newborns. ConclusionsTo reduce false positive and false negative cases, newborn screening programs should be cognizant of factors that influence CK-MM when determining cut-offs for the assay. Variability based on age at specimen collection and birth weight are primarily observed within the first week of life. Therefore, particularly during this time period, multi-tiered cut-offs based on age of collection and lower cut-offs for premature and low birth weight babies are recommended. Other cut-off determinants may include sex, race/ethnicity and seasonal temperature.

Individualized Summary Assessment of Detailed Neuropsychological Testing for the Etiological Diagnosis of Newly Detected Cognitive Impairment in Hospitalized Geriatric Patients.

Neuropsychological testing (NPT) of geriatric inpatients can be affected by the acute illness and/or the hospitalization. To test individualized interpretation of detailed NPT for the differentiation between primary 'neurodegenerative' etiologies (predominantly Alzheimer's disease) and 'other' etiologies (including cerebrovascular disease) of newly detected cognitive impairment in geriatric inpatients without and with delirium in remission. 96 geriatric inpatients (81.9±5.6 years, 64.6% females) with clinically uncertain cognitive impairment were included. 31.3% had delirium in remission that was not considered the primary cause of the cognitive impairment. Categorization of the most likely etiology as 'neurodegenerative' or 'other' was established retrospectively by a study neuropsychologist based on individualized summary assessment of detailed NPT compiled in a standardized vignette. The etiological diagnosis based on FDG-PET served as gold standard (54.2% 'neurodegenerative', 45.8% 'other'). Individualized summary assessment by the study neuropsychologist was correct in 80 patients (83.3%, 8 false positive, 8 false negative). The impact of delirium in remission was not significant (p = 0.237). Individualized summary assessment by an independent neuropsychologist resulted in more false positive cases (n = 22) at the same rate of false negative cases (n = 8). Automatic categorization with a decision tree model based on the most discriminative NPT scores was correct in 68 patients (70.8%, 14 false positive, 14 false negative). Individualized summary assessment of detailed NPT in the context of relevant clinical information might be useful for the etiological diagnosis of newly detected cognitive impairment in hospitalized geriatric patients, also in patients with delirium in remission, but requires task-specific expertise.

Applying Artificial Intelligence to Pediatric Chest Imaging: Reliability of Leveraging Adult-Based Artificial Intelligence Models

ObjectiveThe scarcity of artificial intelligence (AI) applications designed for use in pediatric patients can cause a significant health disparity in this vulnerable population. We investigated the performance of an adult-trained algorithm in detecting pneumonia in a pediatric population to explore the viability of leveraging adult-trained algorithms to accelerate pediatric AI research. MethodsWe analyzed a publicly available pediatric chest x-ray dataset using an AI algorithm from TorchXRayVision. A 60% threshold was used to make binary predictions for pneumonia presence. Predictions were compared with dataset labels. Performance measures including true-positive rate, false-positive rate, true-negative rate, false-negative rate, sensitivity, specificity, positive predictive value (PPV), negative predictive value, accuracy, and F1-score were calculated for the complete dataset and bacterial and viral pneumonia subsets. ResultsOverall (n = 5,856), the algorithm identified 3,923 cases with pneumonia (67.00%) and 1,933 (33.00%) normal cases. In comparison with the actual image labels, there were 3,411 (58.25%) true-positive cases, 512 (8.74%) false-positive cases, 1,071 (18.29%) true-negative cases, and 862 (14.72%) false-negative cases resulting in 79.83% sensitivity, 67.66% specificity, 86.95% PPV, 55.41% negative predictive value, and 76.54% accuracy and an F1-score of 0.83. Although the performance remained consistent in the bacterial pneumonia group, there was a significant decrease in PPV (69.9%) and F1-score (0.74) in the viral pneumonia group. ConclusionAn adult-trained model adequately detected pneumonia in pediatric patients aged 1 to 5 years. Though models trained exclusively on pediatric images performed better, leveraging adult-based algorithms and datasets can expedite pediatric AI research.

FSFNet: Foreground Score-Aware Fusion for 3-D Object Detector Under Unfavorable Conditions

Nowadays, various multimodal fusion based 3D object detectors appear to provide a potential opportunity to solve the failure cases in single modality methods. However, current fusion approaches still face some unfavorable factors, e.g., poor illumination driving conditions and crowded traffic scenarios, which will cause unsatisfying image quality and objects occlusion. To this end, we present a multimodal fusion network FSFNet consisting of local graph-aware point cloud backbone, foreground score-aware fusion network (FSFN), and the proposals refining loss for 3D object detection task in this paper. Concretely, the directed graph is built to generate edge-wise features for each point, and the point features are supplemented with graph information in local graph-aware point cloud backbone. To alleviate the defect of undesirable image quality features caused by poor illumination condition, FSFN is designed to produce an adaptive multi-modal feature by taking point-wise foreground scores into consideration. Hence, level-wise point features with high confidence are fully utilized and the imperfect image information is suppressed in fusion stage. We further introduce proposals refining loss to reduce false positive and false negative cases in crowded scenes by optimizing the location and scores of predicted 3D bounding boxes. Extensive experiments conducted on KITTI benchmark demonstrate that FSFNet owns its superiority over state-of-the-art networks. Moreover, FSFN is verified to be robust against the image inputs under poor illumination conditions.

End-user acceptability of a prototype digital stethoscope to diagnose childhood pneumonia- a qualitative exploration from Sylhet, Bangladesh

BackgroundConsidering the high frequency of respiratory infections among children in low- and middle-income countries (LMICs), the World Health Organization (WHO) developed a pragmatic guideline for managing pneumonia in low-resource settings. The guideline’s low specificity leads to many false-positive pneumonia cases receiving antibiotic treatment. Integrating diagnostic technology to incorporate lung sounds into WHO guidelines could improve childhood pneumonia diagnosis and management. This qualitative study aimed to explore the acceptability of a prototype digital stethoscope device among potential end-users in Bangladesh.MethodsWe conducted four focus group discussions (FGDs) with beneficiaries and service providers who used a 2018 digital stethoscope prototype. The data collection was conducted in November 2020. The study was carried out at Zakiganj Upazila (sub-district) of Sylhet district of Bangladesh. A total of 34 respondents, including parents of under-5 children, Community Health Care Providers (CHCPs), and community leaders were enrolled. Two researchers (TJ and a research assistant (not a co-author of this manuscript) conducted the FGDs. Verbatim transcripts were prepared, and translations were completed. Coding was executed in Microsoft Excel, and relevant quotes were extracted to ascertain the emerging themes. To ensure validity, two researchers coded the dataset independently and inconsistencies were resolved through discussion.FindingsMothers were more aware of the digital stethoscope than fathers. Except for the female community leaders, male leaders were unaware of the stethoscopes. Most CHCPs had positive perceptions of the digital stethoscope. They appreciated stethoscope training as they learned about new technology and diagnostic approaches. The users mentioned several technical shortcomings of the prototype device. A few stakeholders expressed dissatisfaction with the level of community involvement and information sharing from the study. The use of the device plummeted during the COVID-19 pandemic for fear of infection, to counteract which the CHCPs cleaned the device with chlorhexidine after every application as a precaution.ConclusionOverall, device use was supported by stakeholders despite perceptions that the prototype had some technological limitations, community engagement was suboptimal, and the COVID-19 pandemic caused disruptions. Stronger community engagement, addressing technological issues, and further research on its health systems application would improve the acceptability and effective use of the digital stethoscope.

A prospective phase 2 study of expeditious EGFR genotyping and immediate therapeutic initiation through extracellular vesicles (EV)-based bronchoalveolar lavage fluid (BALF) liquid biopsy in advanced NSCLC patients.

In our previous study, epidermal growth factor receptor (EGFR) genotyping using extracellular vesicles (EV)-derived DNA isolated from bronchoalveolar lavage fluid (BALF) was proven to be highly concordant with conventional tissue-based genotyping and its turn-around-time (TAT) was only 1-2 days. On this background, we prospectively validated the performance of EV-based BALF liquid biopsy for EGFR genotyping in the real practice of advanced non-small cell lung cancer (NSCLC) patients. After screening 120 newly diagnosed stage III-IV NSCLC patients, 51 cases were detected as EGFR-mutated by EV-based BALF EGFR genotyping and 40 patients were enrolled for gefitinib treatment. BALF EV were isolated by ultracentrifuge method and EGFR genotyping was performed with PCR-based PNA-clamping assisted fluorescence melting curve analysis. The objective response rate, progression-free survival (PFS), TAT, time to treatment initiation (TTI), and concordance rate were analyzed with clinical parameters. There was only one false positive case among the 120 screened patients and the overall concordance rate between tissue biopsy and EV-based BALF liquid biopsy was 99.2% including the subtype of EGFR mutations. TAT for EV-based BALF EGFR genotyping was 1.9±1.1 days, while tissue-based TAT was 12.1±7.2 days (P<0.001). EGFR genotyping was determined even before obtaining histopathologic report in most cases. TTI in BALF EGFR genotyping was faster than tissue genotyping (7.8±6.5 vs. 13.8±12.9 days). Therapeutic outcomes of response rate and PFS were almost similar to tissue-based results. We demonstrated, for the first time, that EV-based BALF liquid biopsy should be an excellent platform for expeditious EGFR genotyping and rapid therapeutic intervention even before obtaining the result of histopathology in advanced NSCLC patients.