Understanding the variant landscape, and genetic epidemiology of Multiple Endocrine Neoplasia in India.

PURPOSE: Multiple Endocrine Neoplasia (MEN) is a group of familial cancer syndromes that encompasses several types of endocrine tumors differentiated by genetic mutations in RET, MEN1 and CDKN1B genes. Accurate diagnosis of MEN subtypes can thus be performed through genetic testing. However, MEN variants remain largely understudied in Indian populations. Additionally, few dedicated resources to understand these disorders currently exist. METHODS: Using the gold-standard ACMG/AMP guidelines, we systematically classified variants reported across the three genes in the IndiGen dataset, and established the genetic epidemiology of MEN in the Indian population. We further classified ClinVar and Mastermind variants and compiled all into a database. Finally, we designed a multiplex primer panel for rapid variant identification. RESULTS: We have established the genetic prevalence of MEN as the following: 1 in 1026 individuals is likely to be afflicted with MEN linked with pathogenic RET mutations. We have further created the MAPVar database containing 3280 ACMG-classified variants freely accessible at: https://clingen.igib.res.in/MAPVar/ . Finally, our NGS primer panel covers 33 exonic regions across two pools through 38 amplicons with a total amplified region of 65 kb. CONCLUSION: Our work establishes that MEN is a prevalent disorder in India. The rare nature of Indian variants underscores the need of genomic and functional studies to establish a more comprehensive variant landscape. Additionally, our panel offers a means of cost-effective genetic testing, and the MAPVar database a ready reference to aid in a better understanding of variant pathogenicity in clinical as well as research settings.

In vivo confocal Raman spectroscopy: The window into the skin.

Development of in vivo confocal Raman spectroscopy (ICRS) methodology over the last 20 years has enabled previously unavailable capability to acquire molecular concentration gradients across the stratum corneum (SC), at the micron level and in a clinical setting. Professor Tony Rawlings has been a driving force in SC research for over 30 years, working with a wide range of teams across the world. Because a detailed knowledge of skin biochemistry was key to interpreting ICRS-acquired molecular concentration gradients, the authors formed a close working relationship with Professor Rawlings during the development of ICRS. This article, therefore, presents a summary of this process and how challenges raised by application of ICRS were tackled, towards the goal of validating the technique for clinical skin measurement.

Le développement de la méthodologie de spectroscopie confocale Raman in vivo (In vivo Confocal Raman Spectroscopy, ICRS) au cours des 20 dernières années a permis d'acquérir des gradients de concentration moléculaire dans l'ensemble du stratum corneum (SC), au niveau du micron et dans un contexte clinique, ce qui était impossible auparavant. Le professeur Tony Rawlings joue un rôle moteur dans la recherche sur le SC depuis plus de 30 ans et travaille avec de nombreuses équipes à travers le monde. Étant donné qu'une connaissance détaillée de la biochimie cutanée était essentielle à l'interprétation des gradients de concentration moléculaire acquis par l'ICRS, les auteurs ont établi une relation de travail étroite avec le professeur Rawlings pendant le développement de l'ICRS. Cet article présente donc un résumé de ce processus et de la manière dont les défis soulevés par l'application de l'ICRS ont été abordés dans le but de valider la technique de mesure clinique de la peau.

The diagnostic cascade for patients with hepatitis delta infection in France, 2018-2022: A cross-sectional study.

BACKGROUND AND AIMS: Chronic hepatitis D infection is the most severe form of viral hepatitis and can rapidly progress to cirrhosis or hepatocellular carcinoma. Despite recommendations for systematic screening of hepatitis B surface antigen (HBsAg)-positive individuals, data from real-world studies have reported a low frequency of hepatitis D (or delta) virus (HDV) screening. Our cross-sectional analysis evaluated the diagnostic cascade for hepatitis D infection in tertiary centres and described the characteristics of HDV-positive patients. METHODS: A total of 6772 individuals who tested HBsAg positive for the first time between 2018 and 2022 were retrospectively included. Demographic, clinical and laboratory data were analysed. RESULTS: A total of 5748 HBsAg-positive individuals (84.9%) were screened for HDV infection. The screening rate varied from 63% to 97% according to the screening strategy used in the centres including or not HDV reflex testing. The prevalence of HDV infection was 6.3%. HDV RNA levels were determined in 285 of the 364 (78.3%) HDV antibody screening-positive patients, and 167 (58.6%) had active HDV infection. 66.8% were males, with a mean age of 44.9 years. A total of 97.5% were born abroad, and 92.9% were HBeAg negative. At the time of diagnosis, HDV RNA levels were 6.0 Log UI/mL; 60.1% had alanine aminotransferase >40 U/L, and 56.3% had significant fibrosis (≥F2), including 41.6% with cirrhosis. The most common genotype was HDV-1 (75.4%). Coinfections were not uncommon: 7.4% were HIV positive, and 15.0% were HCV antibody positive. CONCLUSIONS: The present study highlights the need for increased screening and monitoring of HDV infection. Reflex testing helps to identify HDV-infected individuals.

The Role of Preoperative Abdominal Ultrasound in the Preparation of Patients Undergoing Primary Metabolic and Bariatric Surgery: A Machine Learning Algorithm on 4418 Patients' Records.

BACKGROUND: The utility of preoperative abdominal ultrasonography (US) in evaluating patients with obesity before metabolic bariatric surgery (MBS) remains ambiguously defined. METHOD: Retrospective analysis whereby patients were classified into four groups based on ultrasound results. Group 1 had normal findings. Group 2 had non-significant findings that did not affect the planned procedure. Group 3 required additional or follow-up surgeries without changing the surgical plan. Group 4, impacting the procedure, needed further investigations and was subdivided into 4A, delaying surgery for more assessments, and 4B, altering or canceling the procedure due to critical findings. Machine learning techniques were utilized to identify variables. RESULTS: Four thousand four hundred eighteen patients' records were analyzed. Group 1 was 45.7%. Group 2, 35.7%; Group 3, 17.0%; Group 4, 1.5%, Group 4A, 0.8%; and Group 4B, 0.7%, where surgeries were either canceled (0.3%) or postponed (0.4%). The hyperparameter tuning process identified a Decision Tree classifier with a maximum tree depth of 7 as the most effective model. The model demonstrated high effectiveness in identifying patients who would benefit from preoperative ultrasound before MBS, with training and testing accuracies of 0.983 and 0.985. It also showed high precision (0.954), recall (0.962), F1 score (0.958), and an AUC of 0.976. CONCLUSION: Our study found that preoperative ultrasound demonstrated clinical utility for a subset of patients undergoing metabolic bariatric surgery. Specifically, 15.9% of the cohort benefited from the identification of chronic calculous cholecystitis, leading to concomitant cholecystectomy. Additionally, surgery was postponed in 1.4% of the cases due to other findings. While these findings indicate a potential benefit in certain cases, further research, including a cost-benefit analysis, is necessary to fully evaluate routine preoperative ultrasound's overall utility and economic impact in this patient population.

Prevention of Ovarian Cancer: Where are We Now and Where are We Going?

PURPOSE OF REVIEW: To describe current and future strategies to reduce the burden of ovarian cancer through prevention. RECENT FINDINGS: Current strategies in genetic testing are missing a substantial number of individuals at risk, representing a missed opportunity for ovarian cancer prevention. Past efforts at screening and early detection have thus far failed to improve ovarian cancer mortality, and novel techniques are needed. Surgical prevention is highly effective, but surgical menopause from oophorectomy has significant side effects. Novel surgical strategies aimed at reducing risk while minimizing these harms are currently being studied. To maximize ovarian cancer prevention, a multi-pronged approach is needed. We propose that more inclusive and accurate genetic testing to identify more individuals at risk, novel molecular screening and early detection, surgical prevention that maximizes quality of life while reducing risk, and broader adoption of targeted and opportunistic salpingectomy will together reduce the burden of ovarian cancer.

Many Paths to the Summit: Survey of Step 1 Study Methods with Pass/Fail Scoring.

Introduction: After the United States Medical Licensing Examination (USMLE) shifted the Step 1 licensure exam to pass/fail, there have been limited studies to analyze changes in student study strategies. Material and Methods: Surveys were distributed to third- and fourth-year medical students at the University of Michigan Medical School (UMMS). The response rate was 66%. Results: The largest proportion (18.81%) of students chose 8 weeks of study time. 40.59% of students increased the length of their study period. To determine time allocated for dedicated study, 37.26% of respondents consulted near-peers who had already taken Step 1. Students also considered prior experiences with standardized tests (15.57%), personal reasons (14.62%), and conversations with school advisors (13.21%). 44.55% of students studied for 9-11 h a day, and 42.57% studied for 5-8 h a day. 52.69% of students scored between 70 and 80% on their final practice NBME test before their Step 1 exam. One hundred percent of respondents passed the exam. Exam non-extenders achieved higher end average practice test scores with shorter study periods. No differences in Step 1 study time or intensity were found when comparing students by intended specialty competitiveness. Conclusion: Our results demonstrated patterns in study strategies for the new pass/fail Step 1 exam that may prove useful for curriculum design and schedule plan for future cohorts. Supplementary Information: The online version contains supplementary material available at 10.1007/s40670-024-02072-2.

Seroreactivity against Leptospira spp. differs between community cats and privately-owned cats in Hong Kong.

Leptospirosis is a bacterial zoonotic disease of major One Health significance and public health impact globally, with a wide host range including mammals, cetaceans and herpetofauna. This study aimed to determine Leptospira seroprevalence, risk factors for seroreactivity and prevalence of urinary Leptospira shedding among domestic cats in Hong Kong. Microagglutination testing of 22 Leptospira serovars from 20 serogroups was performed on 738 sera from outdoor free-roaming "community" cats (n = 391) and privately-owned (n = 347) cats. Urine from 268 community cats was tested for pathogenic Leptospira DNA by qPCR targeting lipL32. Potential risk factors associated with exposure were assessed using logistic regression. Overall Leptospira seroprevalence was 9.35%. Of 14 serogroups detected, Javanica (4.3%), Djasiman (2.3%) and Australis (1.5%) were most common. Seroreactivity was significantly higher among community (13.3%) than privately-owned cats (4.9%; OR 2.98 [95% CI 1.68-5.25], P < 0.001), especially to Javanica (7.65% of community cats versus 0.58% of privately-owned cats (P < 0.001). Antibody titres to all serogroups ranged from 1:100 to 1:6400 (median 1:200) and were highest for Javanica (median 1:800). Leptospira DNA was detected in urine from 12/268 community cats (4.48%; median load 6.42 × 102 copies/mL urine; range 1.40 × 101-9.63 × 104). One in three seroreactive community cats with paired urine and blood samples had leptospiruria. After adjusting for source, none of breed, sex, neuter status, age, district rodent infestation rate, serum alanine transaminase or creatinine values were associated with seroreactivity. Cats in Hong Kong are exposed to a diversity of Leptospira serogroups and can shed Leptospira silently in urine. The higher seroprevalence among outdoor free-roaming community cats highlights the importance of environmental drivers in leptospirosis transmission and risks of exposure for sympatric human populations. Gloves should be worn when handling feline urine to minimise the risk of zoonotic transmission from subclinically infected cats.

C-reactive protein point-of-care testing in primary care-broader implementation needed to combat antimicrobial resistance.

This study presents the perspective of an international group of experts, providing an overview of existing models and policies and guidance to facilitate a proper and sustainable implementation of C-reactive protein point-of-care testing (CRP POCT) to support antibiotic prescribing decisions for respiratory tract infections (RTIs) with the aim to tackle antimicrobial resistance (AMR). AMR threatens to render life-saving antibiotics ineffective and is already costing millions of lives and billions of Euros worldwide. AMR is strongly correlated with the volume of antibiotics used. Most antibiotics are prescribed in primary care, mostly for RTIs, and are often unnecessary. CRP POCT is an available tool and has been proven to safely and cost-effectively reduce antibiotic prescribing for RTIs in primary care. Though established in a few European countries during several years, it has still not been implemented in many European countries. Due to the complexity of inappropriate antibiotic prescribing behavior, a multifaceted approach is necessary to enable sustainable change. The effect is maximized with clear guidance, advanced communication training for primary care physicians, and delayed antibiotic prescribing strategies. CRP POCT should be included in professional guidelines and implemented together with complementary strategies. Adequate reimbursement needs to be provided, and high-quality, and primary care-friendly POCT organization and performance must be enabled. Data gathering, sharing, and discussion as incentivization for proper behaviors should be enabled. Public awareness should be increased, and healthcare professionals' awareness and understanding should be ensured. Impactful use is achieved when all stakeholders join forces to facilitate proper implementation.

Remote self-testing for adult cochlear implant users: the effect of wireless streaming on speech recognition in noise.

OBJECTIVES: Wireless sound transfer methods for cochlear implant sound processors have become popular for remote self-assessed hearing tests. The aim of this study was to determine (1) spectral differences in stimuli between different wireless sound transfer options and (2) the effect on outcomes of speech recognition tests in noise. DESIGN: In study 1, the frequency response of different streaming options (Phonak Roger Select, Cochlear Mini Mic 2+, telecoil and Bluetooth connection) was measured by connecting headphones to CI sound processors. Study 2 followed a repeated measures design in which digits-in-noise (DIN) tests were performed using wireless streaming to sound processors from Cochlear, Advanced Bionics, and MED-EL. STUDY SAMPLE: 20 normal hearing participants. RESULTS: Differences in frequency response between loudspeaker and wireless streaming conditions were minimal. We did not find significant difference in DIN outcome (F4,194 = 1.062, p = 0.376) between the wireless transfer options with the Cochlear Nucleus 7 processor. No significant difference in DIN outcomes was found between Bluetooth streaming and the loudspeaker condition for all of the three tested brands. The mean standard error of measurement was 0.72 dB. CONCLUSIONS: No significant differences in DIN test outcomes between wireless sound transfer and the reference method were found.

Novel truncating germline variant reinforces TINF2 as a susceptibility gene for familial non-medullary thyroid cancer.

BACKGROUND: It has long been observed that there are families in which non-medullary thyroid cancer (NMTC) occurs, but few syndromes and genes have been described to date. Proteins in the shelterin complex have been implied in cancer. Here, we have studied shelterin genes in families affected by NMTC (FNMTC). METHODS: We performed whole-exome sequencing (WES) in 10 affected individuals from four families with at least three affected members. Polymerase chain reaction (PCR) and Sanger sequencing were performed to search for variants in the TINF2 gene in 40 FNMTC families. TINF2 transcripts and loss of heterozygosity (LOH) were studied in several affected patients of one family. RESULTS: We found the c.507G>T variant in heterozygosis in the TINF2 gene in one family, co-segregating in all five affected members. This variant affects the normal splicing. LOH was not observed. CONCLUSIONS: Our results reinforce the TINF2 gene as a susceptibility cause of FNMTC suggesting the importance of location of frameshift variants in TINF2. According to our data and previous literature, TINF2 pathogenic variants appear to be a significant risk factor for the development of NMTC and/or melanoma.

Comprehensive analysis of chemical and enantiomeric stability of terpenes in Cannabis sativa L. flowers.

OBJECTIVE: Cannabis sativa L. is renowned for its medicinal and recreational uses. With the increasing global legalization of C. sativa L.-based products for medicinal purposes, there is a growing need for well-characterized products. While the stability of cannabinoids such as tetrahydrocannabinol and cannabidiol is well understood, information on the chemical and enantiomeric stability of terpenes remains scarce. This is despite the fact that terpenes are also thought to have pharmacological activity and may contribute to the overall effect of C. sativa L. METHODS: To address these challenges, four analytical methods based on chiral, polar, and apolar gas chromatographic separation combined with either MS or FID detection were developed and validated. These methods successfully separated and quantified a total of 29 terpenes, including 13 enantiomers and 5 diastereomers specific to C. sativa L. Furthermore, terpenes and authentic C. sativa L. flowers and extracts were subjected to UV and heat treatments to observe potential degradation reactions over time. RESULTS: Each terpene generates a unique pattern of degradation products resulting in a diverse array of oxidation and cyclization products. P-cymene was identified as a major product of terpene aging. Notably, no enantiomeric conversion was detected, suggesting that the formation of (-)-α-pinene in cannabis extracts, for example, originates from other terpenes. CONCLUSION: Terpenes have different degradation rates, even though they are structurally similar. In addition, cultivar- and growth-condition-specific enantiomeric ratios were observed in C. sativa L., confirming that enantiomer production is species-specific and has to be considered for therapeutical applications.

Evaluation of spectroscopic techniques for on-site drug testing of festival seizures.

Despite the fact that drugs of abuse are illegal, a drug-free festival still remains an utopia in most settings. For law enforcement purposes, it is necessary to rapidly determine whether controlled substances are involved. On-site testing is a challenging task because drugs appear in different physical forms and concentrations. The aim of this study was to compare the performance of two spectroscopic techniques, Raman and Fourier transform-infrared (FT-IR), for the testing of drug seizures at a dance festival. First, samples were measured through packaging with Raman. Subsequently, homogenized samples were analysed with FT-IR. For MDMA tablets, a chemometric model was applied on the FT-IR spectra for the dose estimation. After the festival, results were confirmed in the forensic laboratory with gas chromatography coupled with mass spectrometry (GC-MS) and gas chromatography with flame ionization detection (GC-FID). In total, 166 samples of which 90 tablets, 53 powders, 16 crystals and 7 liquids were analysed. MDMA, cocaine and ketamine were the top three drugs seized. The Raman technique was suitable for powders and crystals (sensitivity of 100% and 81%, respectively). However, in comparison with FT-IR, Raman performance was lower for the analysis of liquids (sensitivity of 67%) and 'ecstasy'-like tablets (sensitivity of 41%). Overall, sensitivities above 95% were obtained with FT-IR. The MDMA doses of the tablets, determined on-site, ranged between 52 mg and 336 mg MDMA hydrochloride. For a quick identification of a variety of drugs on-site, the combination of Raman and FT-IR is recommended. It should be emphasized that optimized settings, in-house libraries and analysis by trained operators are essential to obtain correct results.

Lab-in-a-Vial Rapid Test for Internet of Things-Embedded Point-of-Healthcare Protein Biomarker Detection in Bodily Fluids.

Amateurs often struggle with detecting and quantifying protein biomarkers in body fluids due to the high expertise required. This study introduces a Lab-in-a-Vial (LV) rapid diagnostic platform, featuring hydrangea-like platinum nanozymes (PtNH), for rapid, accurate detection and quantification of protein biomarkers on-site within 15 min. This method significantly enhances detection sensitivity for various biomarkers in body fluids, surpassing traditional methods such as enzyme-linked immunosorbent assays (ELISA) and lateral flow assays (LFA) by ≈250 to 1300 times. The LV platform uses a glass vial coated with specific bioreceptors such as antigens or antibodies, enabling rapid in vitro evaluation of disease risk from small fluid samples, similar to a personal ELISA-like point-of-care test (POCT). It overcomes challenges in on-site biomarker detection, allowing both detection and quantification through a portable wireless spectrometer for healthcare internet of things (H-IoT). The platform's effectiveness and adaptability are confirmed using IgG/IgM antibodies from SARS-CoV-2 infected patients and nuclear matrix protein (NMP22) from urothelial carcinoma (UC) patients as biomarkers. These tests demonstrated its accuracy and flexibility. This approach offers vast potential for diverse disease applications, provided that the relevant protein biomarkers in bodily fluids are identified.

Nanoindentation creep: The impact of water and artificial saliva storage on milled and 3D-printed resin composites.

PURPOSE: This study evaluated the effects of artificial saliva and distilled water on the nanoindentation creep of different 3D-printed and milled CAD-CAM resin composites. MATERIAL AND METHODS: Disk-shaped specimens were subtractively fabricated from polymer-infiltrated ceramic network (EN) and reinforced resin composite (B) and additively from resin composite (C) and hybrid resin composite (VS) using digital light processing (DLP). Specimens from each material were divided into two groups according to their storage conditions (artificial saliva or distilled water for 3 months). Creep was analyzed by nanoindentation testing. Statistical analysis was done using two-way ANOVA, one-way ANOVA, Bonferroni post hoc tests, and independent t-test (α = 0.05). RESULTS: The main effects of material and storage conditions, and their interaction were statistically significant on nanoindentation (p < 0.001). Storage condition had the greatest influence (partial eta squared ηP 2 = 0.370), followed by the material (ηP 2 = 0.359), and the interaction (ηP 2 = 0.329). The nanoindentation creep depths after artificial saliva storage ranged from 0.34 to 0.51 µm and from 0.50 to 0.87 µm after distilled water storage. One of the additively manufactured groups had higher nanoindentation creep depths in both storage conditions. CONCLUSIONS: All specimens showed comparable performance after artificial saliva storage, but increased nanoindentation creep after distilled water storage for 3 months. The subtractive CAD-CAM blocks showed superior dimensional stability in terms of nanoindentation creep depths in both storage conditions. Additively manufactured composite resins had lower dimensional stability than one of the subtractively manufactured composites, which was demonstrated as having higher creep deformation and maximum recovery. However, after artificial saliva storage, one of the additively manufactured resins had dimensional stability similar to that of subtractively manufactured.

Improving Private Well Testing Programs: Experimental Evidence from Iowa.

Approximately 23 million U.S. households rely on private wells for drinking water. This study first summarizes drinking water behaviors and perceptions from a large-scale survey of households that rely on private wells in Iowa. Few households test as frequently as recommended by public health experts. Around 40% of households do not regularly test, treat, or avoid their drinking water, suggesting pollution exposure may be widespread among this population. Next, we utilize a randomized control trial to study how nitrate test strips and information about a free, comprehensive water quality testing program influence households' behaviors and perceptions. The intervention significantly increased testing, including high-quality follow-up testing, but had limited statistically detectable impacts on other behaviors and perceptions. Households' willingness to pay for nitrate test kits and testing information exceeds program costs, suggesting that the intervention was welfare-enhancing.

Evaluation of EUCAST rapid antimicrobial susceptibility test directly from positive blood culture for Pseudomonas aeruginosa.

In this study, we evaluated the performance of the EUCAST RAST method on a collection of 154 clinical strains of P. aeruginosa, including strains resistant to ceftazidime and carbapenems. While the test is convenient for routine laboratories, we observed significant rates of VME (ranging from 0.0 to 15.0%) and ME (ranging from 1.3 to 16.3%) after 6 h, particularly for key antibiotics such as ceftazidime, piperacillin/tazobactam, and meropenem. Extending the incubation time to 8 h may improve results (CA ranging from 87.2 to 99%), but caution is required in interpretation due to persistence of VME (ranging from 0.0 to 15.6%) and ME (ranging from 0.0 to 11.7%).

Fetal endocrine axes mRNA expression levels are related to sex and intrauterine position.

BACKGROUND: The hypothalamic-pituitary-adrenal (HPA) and -gonadal (HPG) axes are two major pathways that connect the neural and endocrine systems in vertebrates. Factors such as prenatal stress and maternal exposure to exogenous steroids have been shown to affect these pathways during fetal development. Another less studied factor is the transfer of hormones across fetuses in multifetal pregnancies. This form of transfer has been shown to influence the morphology, anatomy, physiology, and behavior of the offspring in litter-bearing mammals, an influence termed the intrauterine position (IUP) effect. In this study, we sought to delineate how the IUP effects HPA and HPG brain receptors, peptides, and enzymes (hereafter components) in utero and how these influences may differ between males and females. METHODS: We utilized the unconventional model of culled free-ranging nutria (Myocastor coypus), with its large natural variation. We collected brain tissues from nutria fetuses and quantified the expression of key HPA and HPG components in three brain regions: prefrontal cortex, hypothalamus, and striatum. RESULTS: We found an interaction between sex and IUP in the mineralocorticoid receptor (MR), gonadotropin-releasing hormone receptor (GNRHR), androgen receptor (AR), and estrogen receptor alpha (ESR1). IUP was significant in both gonadotropin-releasing hormone (GnRH) and its receptor GNRHR, but in different ways. In the hypothalamus, fetuses adjacent to same-sex neighbors had higher expression of GnRH than fetuses neighboring the opposite sex. Conversely, in the cortex, GNRHR exhibited the inverse pattern, and fetuses that were neighboring the opposite sex had higher expression levels than those neighboring the same sex. Regardless of IUP, in most components that showed significant sex differences, female fetuses had higher mRNA expression levels than male fetuses. We also found that HPA and HPG components were highly related in the early stages of gestation, and that there was an interaction between sex and developmental stage. In the early stages of pregnancy, female component expression levels were more correlated than males', but in the last trimester of pregnancy, male components were more related to each other than female's. CONCLUSIONS: This study suggests that there are sexually different mechanisms to regulate the HPA and HPG axes during fetal development. Higher mRNA expression levels of endocrine axes components may be a mechanism to help females cope with prolonged androgen exposure over a long gestational period. Additionally, these findings suggest different coordination requirements of male and female endocrine axes during stages of fetal development.

3D-printed neck phantoms with detailed anatomy for ultrasound-guided procedure and device testing.

Objectives: With rapid advances in ultrasound-guided procedures, there is an unmet need for echogenic phantoms with sufficient anatomical details for artificial intelligence and ultrasound-guided device testing. We developed a method for creating neck phantoms for novel otolaryngology-related device testing. To achieve accurate representation of the anatomy, we utilized CT scans and 3D printing technology to create customized agar molds, thus providing high-fidelity yet cost-effective tools. Methods: Based on previous studies, the key components in our neck phantom include the cervical vertebrae, trachea, common carotid arteries, internal jugular veins, thyroid gland, and surrounding soft tissue. Open-source image analysis software were employed to process CT data to generate high fidelity 3D models of the target structures. Resin molds were 3D printed and filled with various agar mixtures to mimic anatomical echogenicity. Results: Following the method proposed, we successfully assembled the neck phantom which provided a detailed representation of the target structures. To evaluate the results, ultrasound data was collected on the phantom and living tissue and analyzed with ImageJ. We were able to demonstrate echogenicity comparable to that of living tissue. Conclusion: The proposed method for building neck phantoms with detailed anatomical features offers a valuable, detailed, low-cost tool for medical training and device testing in otolaryngology, particularly for novel devices that involve artificial intelligence (AI) guidance and robotic-based needle insertion. Additional anatomical refinements and validation studies could further enhance the consistency and accuracy, thus paving the way for future advancements in ultrasound training and research, and ultimately benefiting patient care and safety.

Discordance Between Phenotypic and WGS-Based Drug Susceptibility Testing Results for Some Anti-Tuberculosis Drugs: A Snapshot Study of Paired Mycobacterium tuberculosis Isolates with Small Genetic Distance.

Background: Current tuberculosis treatment regimens primarily rely on phenotypic drug susceptibility testing and rapid molecular assays. Although whole-genome sequencing (WGS) offers a promising alternative, disagreements between phenotypic and molecular testing methods remain. In this retrospective study, we compared the phenotypic and WGS-predicted drug resistance profiles of paired Mycobacterium tuberculosis isolates with small genetic distances (≤10 single nucleotide variants) obtained from patients with longitudinal single-episode or recurrent tuberculosis. Additionally, we investigated the distribution of drug-resistance-conferring variants among the identified M. tuberculosis genotypes. Methods: Paired M. tuberculosis isolates from 46 patients with pulmonary tuberculosis (2002-2019) were analyzed. Spoligotyping was performed for all the isolates. WGS data were processed using TB-Profiler software to genotype the strains and detect variants in M. tuberculosis genes associated with drug resistance. The significance of these variants was evaluated using the M. tuberculosis variant catalog developed by the World Health Organization. Phenotypic drug susceptibility test results were obtained from patients' medical records. Results: Among the 46 isolate pairs, 25 (54.3%) harbored drug-resistance-associated variants, with 20 demonstrating identical WGS-predicted drug resistance profiles. Drug-resistant isolate pairs belonged to Lineages 2 and 4, with the most common sub-lineages being 2.2.1 (SIT1 and SIT190 spoligotypes), and 4.3.3 (SIT42). Agreement between phenotypic and WGS-based drug susceptibility testing was highest (>90%) for rifampicin, isoniazid, ethambutol, fluoroquinolones, streptomycin, and amikacin when calculated for M. tuberculosis isolates or isolate pairs. In most discordant cases, isolate pairs harbored variants that could cause low- or moderate-level resistance or were previously associated with variable minimum inhibitory concentrations. Notably, such discrepancies mostly occurred in one isolate from the pair. In addition, differences in resistance-related variant distributions among M. tuberculosis genotypes were observed for most of the analyzed drugs. Conclusion: The simultaneous performance of phenotypic and WGS-based drug susceptibility testing creates the most accurate drug resistance profile for M. tuberculosis isolates and eliminates important limitations of each method.

Sparse testing designs for optimizing predictive ability in sugarcane populations.

Sugarcane is a crucial crop for sugar and bioenergy production. Saccharose content and total weight are the two main key commercial traits that compose sugarcane's yield. These traits are under complex genetic control and their response patterns are influenced by the genotype-by-environment (G×E) interaction. An efficient breeding of sugarcane demands an accurate assessment of the genotype stability through multi-environment trials (METs), where genotypes are tested/evaluated across different environments. However, phenotyping all genotype-in-environment combinations is often impractical due to cost and limited availability of propagation-materials. This study introduces the sparse testing designs as a viable alternative, leveraging genomic information to predict unobserved combinations through genomic prediction models. This approach was applied to a dataset comprising 186 genotypes across six environments (6×186=1,116 phenotypes). Our study employed three predictive models, including environment, genotype, and genomic markers as main effects, as well as the G×E to predict saccharose accumulation (SA) and tons of cane per hectare (TCH). Calibration sets sizes varying between 72 (6.5%) to 186 (16.7%) of the total number of phenotypes were composed to predict the remaining 930 (83.3%). Additionally, we explored the optimal number of common genotypes across environments for G×E pattern prediction. Results demonstrate that maximum accuracy for SA ( ρ = 0.611 ) and for TCH ( ρ=0.341 ) was achieved using in training sets few (3) to no common (0) genotype across environments maximizing the number of different genotypes that were tested only once. Significantly, we show that reducing phenotypic records for model calibration has minimal impact on predictive ability, with sets of 12 non-overlapped genotypes per environment (72=12×6) being the most convenient cost-benefit combination.