High Specificity Research Articles

Isolation and Characterization of Cytotoxic Compounds from Detarium microcarpum Guill. and Perr. Stem Bark.

Globally, about 8.2 million cancer-related deaths are recorded annually. Sadly, most of the deaths result from the toxicity of most chemotherapeutic agents. Hence, there are growing demands for chemotherapeutic agents with high specificity and selectivity. This study was designed to assess the cytotoxic potential of Detarium microcarpum and isolate cytotoxic compounds with better selectivity profiles. Detarium microcarpum Stem bark (DMS) was collected and authenticated at the Forest Herbarium Ibadan (FHI), and a voucher (FHI-111954) was issued. Dried DMS was pulverized and extracted into 70% methanol. The extract was partitioned into hexane, dichloromethane, and ethyl acetate fractions. The cytotoxicities of the extract, fractions, and isolated compounds were determined. The cytotoxicity of the isolated compounds was tested against different cell lines, including human breast (AU565 and MDA MB231), oral adenosquamous (CAL27), and cervical (HeLa) cancer cells, as well as healthy (3T3) non-cancer cells. Methyl gallate, eriodictyol, quercetin, quebrachitol, catechin, catechin gallate, and gallic acid, isolated from dichloromethane and ethyl acetate fractions, displayed weak cytotoxicity against breast (AU565 and MDAMD-231) and cervical (HeLa) cancer cell lines. Interestingly, all the compounds, except gallic acid (48.91±4.51% inhibition), displayed potent cytotoxicity on oral cancer cells. Methyl gallate and quercetin displayed the highest activity, with IC50 values of 89.57±1.98µM and 78.19±1.49µM, respectively. Interestingly, all the compounds were not toxic to healthy non-cancer (3T3) cells. The compounds displayed anticancer activity specific to oral cancer cells and were highly selective for cancer cells without causing significant toxicity to healthy non-cancer cells.

Privacy-preserving large language models for structured medical information retrieval

Most clinical information is encoded as free text, not accessible for quantitative analysis. This study presents an open-source pipeline using the local large language model (LLM) “Llama 2” to extract quantitative information from clinical text and evaluates its performance in identifying features of decompensated liver cirrhosis. The LLM identified five key clinical features in a zero- and one-shot manner from 500 patient medical histories in the MIMIC IV dataset. We compared LLMs of three sizes and various prompt engineering approaches, with predictions compared against ground truth from three blinded medical experts. Our pipeline achieved high accuracy, detecting liver cirrhosis with 100% sensitivity and 96% specificity. High sensitivities and specificities were also yielded for detecting ascites (95%, 95%), confusion (76%, 94%), abdominal pain (84%, 97%), and shortness of breath (87%, 97%) using the 70 billion parameter model, which outperformed smaller versions. Our study successfully demonstrates the capability of locally deployed LLMs to extract clinical information from free text with low hardware requirements.

Bridging Prenatal Diagnostics and AI: A Systematic Review and Meta-Analysis of the Efficacy of Advanced Algorithms in Identifying Congenital Fetal Abnormalities

Congenital fetal abnormalities have emerged as a major cause of infant mortality globally. About 3% of pregnancies are reported as fetal structural anomaly through ultrasound, ranging from minor defect to severe organ system anomalies. Our study aimed to evaluate effectiveness of Artificial intelligence (AI) algorithms in prediction of fetal heart and brain abnormalities by using meta-analysis approach. In this study, the “Reporting Items for Systematic Review and Meta-Analysis (PRISMA)" guidelines were applied for screening and selection of research articles. We searched the research articles according to research aims from Google scholar, PubMed, and Ovid MEDLINE. The data search was limited to January 2015 to May 2024. Two researchers independently screened the studies to detect research aim oriented studies. After screening of titles, those researchers assessed all abstract for eligibility. The risk bias of included studies was assessed by two researchers who used Cochrane library tool (version 5.4.0). Using the DerSimonian-Laird technique, a bivariate random effect model that generated SROC plots, we pooled test estimates. This method is particularly helpful in visualizing variability in the sensitivity and specificity across studies, accounting for the heterogeneity. This improves the reliability of the SROC plot, offering a clear summary of diagnostic performance Using the software RevMan 5.4, we carried out every statistical analysis, particularly heterogeneity through the Q test. About 243,456 screened fetus or pregnant women of 11-32 gestational weeks through 10 studies based on AI algorithms (machine learning, deep learning and AI diagnostic tool). The imaging protocols including ultrasound and MRI were used to take visuals of fetal brain and heart abnormalities in all of including studies. DL and ML algorithms provided high-performance diagnostic predictions, mostly include CNN models that assist in providing accurate diagnostic results with high sensitivity and specificity. The AI algorithm model showed a generally good accuracy scores for detection of brain and heart abnormalities and reference machine learning findings, with sensitivity ranging from 82 to 99% and specificity from 78 to 99%. Fig. 5's area under the SROC curve was 0.960. In contrast to the general detection, ML and DL seems to be more sensitive in diagnosing fetal brain and heart abnormalities. Our study provided the scientific evidence that AI models including deep learning and Machine learning are effective in generating highly reliable detection or classification results for diagnosis of fetal brain and heart anomalies. These strategies can help in reducing infant mortalities, improving treatment outcomes, and postpartum outcomes among women.

Antimicrobial susceptibility testing of bone and joint pathogens using isothermal microcalorimetry

The rise in osteomyelitis and periprosthetic joint infections, in combination with increasing life expectancy and the prevalence of diabetes, underscores the urgent need for rapid and accurate diagnostic tools. Conventional culture‐based methods are often time‐consuming and prone to false‐negatives, leading to prolonged and inappropriate antibiotic treatments. This study aims to improve osteomyelitis diagnostics by decreasing the time to detection and the time to an antibiotic susceptibility result to enable a targeted treatment using isothermal microcalorimetry (IMC). IMC measures heat flow in real‐time, providing insights into bacterial metabolism without the need for labeling. Using clinical isolates from bone infections, assessing their response to antibiotics through IMC, we demonstrated that IMC could detect bacteria within 4 h and determine antimicrobial susceptibility profiles within 2–22 h (median 4.85, range 1.28–21.78). This is significantly faster than traditional methods. A decision tree, based on antibiotic susceptibility, accurately categorized pathogens, achieving high accuracy (74–100%), sensitivity (100%), and specificity (65–100%). These findings suggest that IMC could redefine diagnostics of bone and joint infections and potentially infections in general, offering timely and precise treatment guidance, thereby improving patient outcomes and reducing health care burdens. Further optimization and clinical validation are needed to fully integrate IMC into routine diagnostics.

Corneal Epithelial Wavefront Error as a Novel Diagnostic Marker for Epithelial Basement Membrane Dystrophy

Synopsis: Corneal epithelial wavefront error and epithelial thickness variance qualify as highly sensitive and specific biomarkers for epithelial basement membrane dystrophy (EBMD). The biomarkers show a normalization after treatment of EBMD with phototherapeutic keratectomy. Purpose: To gauge the diagnostic value of epithelial basement membrane dystrophy (EBMD), a novel spectral-domain optical coherence tomography (SD-OCT)-based imaging modality for simultaneous morphological (thickness profile) and refractive (optical wavefront) assessment of the corneal epithelial layer in one of the most common but often underdiagnosed corneal dystrophies. Methods: In this prospective observational study, a total of 32 eyes of 32 patients diagnosed with EBMD and 32 eyes of 32 healthy control subjects were examined with high-resolution anterior segment SD-OCT (MS-39; CSO, Florence, Italy). Various epithelial thickness and epithelial wavefront-derived terms were compared between groups and receiver operating characteristic (ROC) curves were computed to analyze the diagnostic capacity of the respective parameters. A total of 17 of 32 EBMD patients underwent treatment with phototherapeutic keratectomy (PTK) and were followed up for 3 months. Results: Epithelial thickness variance (60.4 ± 56.7 µm versus 7.6 ± 6.1 µm) and interquartile range (11.0 ± 6.9 versus 3.3 ± 1.9 µm) were markedly elevated in EBMD patients as compared with healthy controls (both with p < 0.001). Epithelial wavefront analysis showed a highly statistically significant excess in all examined aberration terms in EBMD patients (all with p < 0.001). Significantly greater areas under the curve (AUCs) were yielded by the epithelial wavefront-derived parameters (e.g., total epithelial wavefront error: AUC = 0.966; 95% confidence interval (CI) 0.932–1) than by the epithelial thickness-derived parameters (e.g., variance: AUC = 0.919; 95% CI 0.848–0.990). Conclusions: Corneal epithelial wavefront aberrometry proved valuable as an objective biomarker for EBMD, with high sensitivity and specificity. PTK resulted in a reduction of morphological and refractive epithelial irregularities in EBMD.

Synthesis of the O antigen repeating units of Escherichia coli serotypes O117 and O107

Abstract Escherichia coli serotype O117 (ECO117) are pathogenic bacteria that produce Shiga toxin. Repeating units of the O antigen of ECO117 have the pentasaccharide structure [4-D-GalNAcβ1-3-L-Rhaα1-4-D-Glcα1-4-D-Galβ1-3-D-GalNAcα1-]n. The related non-pathogenic serotype (ECO107) contains a GlcNAc residue instead of Glc in the repeating unit, and the biosynthetic enzymes involved are almost identical. We assembled these repeating units based on GalNAcα-diphosphate-phenylundecyl (GalNAcα-PP-PhU), an analog of the natural intermediate GalNAc-diphosphate-undecaprenyl. We previously characterized α1,4-Glc-transferase WclY from ECO117 that transfers the Glc residue to Galβ1–3GalNAcα-PP-PhU and showed that Arg194Cys mutants of WclY are active α1,4-GlcNAc-transferases. In this work, the reaction products of WclY were used as acceptor substrates for the final enzymes in pathway, L-Rha-transferase WclX, and GalNAc-transferase WclW, demonstrating a complete synthesis of the ECO117 and O107 repeating units. WclX transfers L-Rha with high specificity for the WclY enzyme product as the acceptor and for TDP-L-Rha as the donor substrate. A number of highly conserved sequence motifs were identified (DDGSxD, DxDD, and YR). Mutational analysis revealed several Asp residues are essential for the catalysis of L-Rha transfer, while mutations of Asp44 and R212 substantially reduced the activity of WclX. WclW is a GT2 enzyme specific for UDP-GalNAc but with broad specificity for the acceptor substrate. Using L-Rhaα-p-nitrophenyl as an acceptor for WclW, the reaction product was analyzed by NMR demonstrating that GalNAc was transferred in a β1–3 linkage to L-Rha. The in vitro synthesis of the repeating units allows the production of vaccine candidates and identifies potential targets for inhibition of O antigen biosynthesis.

High specificity of engineered T cells with third generation CAR (CD28-4-1BB-CD3-ζ) based on biotin-bound monomeric streptavidin for potential tumor immunotherapy

IntroductionImmunotherapy has revolutionized cancer treatment, and Chimeric Antigen Receptor T cell therapy (CAR-T) is a groundbreaking approach. Traditional second-generation CAR-T therapies have achieved remarkable success in hematological malignancies, but there is still room for improvement, particularly in developing new targeting strategies. To address this limitation, engineering T cells with multi-target universal CARs (UniCARs) based on monomeric streptavidin has emerged as a versatile approach in the field of anti-tumor immunotherapy. However, no studies have been conducted on the importance of the intracellular signaling domains of such CARs and their impact on efficiency and specificityMethodHere, we developed second-generation and third-generation UniCARs based on an extracellular domain comprising an affinity-enhanced monomeric streptavidin, in addition to CD28 and 4-1BB co-stimulatory intracellular domains. These UniCAR structures rely on a biotinylated intermediary, such as an antibody, for recognizing target antigens. In co-culture assays, we performed a functional comparison between the third-generation UniCAR construct and two second-generation UniCAR variants, each incorporating either the CD28 or 4-1BB as co-stimulatory domainResultsWe observed that components in culture media could inhibit the binding of biotinylated antibodies to monomeric streptavidin-CARs, potentially compromising their efficacy. Furthermore, third-generation UniCAR-T cells showed robust cytolytic activity against cancer cell lines upon exposure to specific biotinylated antibodies like anti-CD19 and anti-CD20, underscoring their capability for multi-targeting. Importantly, when assessing engineered UniCAR-T cell activation upon encountering their target cells, third-generation UniCAR-T cells exhibited significantly enhanced specificity compared to second-generation CAR-T cellsDiscussionFirst, optimizing culture conditions would be essential before deploying UniCAR-T cells clinically. Moreover, we propose that third-generation UniCAR-T cells are excellent candidates for preclinical research due to their high specificity and multi-target anti-tumor cytotoxicity

Diagnostic performance of rapid on‐site evaluation during bronchoscopy for lung cancer: A comprehensive meta‐analysis

AbstractBackgroundLung cancer is the leading cause of cancer‐related mortality worldwide. Screening high‐risk populations for lung cancer with low‐dose computed tomography (LDCT) reduces lung cancer mortality. Bronchoscopy is a diagnostic procedure used to monitor patients suspected of having lung cancer after LDCT. Rapid on‐site evaluation (ROSE) can improve the diagnostic accuracy of endobronchial ultrasound–guided transbronchial needle aspiration (EBUS‐TBNA), although its diagnostic value remains unclear. In this meta‐analysis, the authors evaluated the diagnostic accuracy of ROSE during bronchoscopy.MethodsThe PubMed, Embase, and Cochrane Library databases were searched for studies evaluating the diagnostic accuracy of ROSE for lung cancer during bronchoscopy. Studies evaluating the performance of ROSE and articles providing sufficient data for constructing a 2 × 2 table on a per‐lesion basis were included. A meta‐analysis was conducted using a bivariate random‐effects model.ResultsIn total, 32 studies involving 8243 lung lesions were included with a pooled sensitivity of 91.8% and a pooled specificity of 94.9%. Subgroup analysis of 12 studies involving 2929 specimens from patients who underwent computed tomography revealed a pooled sensitivity of 93.8% and a pooled specificity of 96%. Further subgroup analysis of seven studies on the diagnostic outcomes of ROSE for intrathoracic or mediastinal lymph nodes through EBUS‐TBNA for lung cancer staging revealed a pooled sensitivity of 90.1% and a pooled specificity of 96.9%.ConclusionsROSE exhibited high sensitivity and specificity for diagnosing lung cancer during bronchoscopy. It also exhibited high sensitivity in detecting lung cancer in patients undergoing LDCT and higher specificity for nodal staging with EBUS‐TBNA.

Intra- and Post-procedural Patient-reported Experience Measures and their Correlation with post-ERCP Adverse Events and Unplanned Healthcare Utilization

Background and Study Aims Post-endoscopic retrograde cholangiopancreatography (ERCP) adverse events (AEs) are common, as is unplanned healthcare utilization (UHU). We aimed to elucidate potential associations between intra- and post-procedural patient-reported experience measures (PREMs) and post-ERCP AEs and UHU. Patients and Methods Prospective data from a multi-center collaborative were used. A validated 0-10 Likert-based PREM assessing intra- and post-procedural symptoms was applied to patients following ERCP and protocolized follow-up was performed at 30 days to identify AEs and UHU for reasons not meeting the definitions of any AE. Multivariable logistic regression was conducted using PREM domains as exposures and individual AEs and UHU as outcomes, with a priori selected patient- and procedure-related covariates. Test performance characteristics and odds ratios (ORs) and 95% confidence intervals (95% CIs) for each PREM domain were reported. Results From September 2018 through October 2023, 3,434 ERCPs were included. Post-procedural abdominal pain of >3 was predictive of pancreatitis (OR 3.71, 2.37-5.73), while a score >6 was strongly predictive of perforation (OR 9.54, 1.10-59.37). Post-procedural pain was also predictive of UHU within 30 days when used as a continuous predictor (OR 1.08 per point, 1.01-1.16). Post-procedural pain of >6 demonstrated high negative predictive values and specificities for post-ERCP AEs. Conclusions Patient-reported symptom scores from a simple Likert-based PREM at the time of discharge from ERCP are associated with presentations for pancreatitis, perforation, and UHU within 30 days. Applying PREMs post-ERCP could potentially prevent UHU and/or facilitate earlier management and improved outcomes for patients with post-ERCP AEs.

The expression and clinical significance of syncytin-1 in serum exosomes of hepatocellular carcinoma patients

Abstract This study aimed to investigate the expression and clinical significance of syncytin-1 in the serum exosomes of hepatocellular carcinoma (HCC) patients. Serum samples were collected from 61 patients with newly diagnosed HCC and 61 healthy individuals. Exosomes were extracted from serum samples and identified using transmission electron microscopy and Western blot. The relative expression levels of syncytin-1 in exosomes were determined by real-time quantitative PCR. The protein expression levels of alpha-fetoprotein and syncytin-1 in HCC patients were detected using enzyme-linked immunosorbent assay (ELISA). Statistical analysis was performed to evaluate the sensitivity and specificity of serum exosomal syncytin-1 in diagnosing HCC. The relationships between syncytin-1 expression and clinical pathological features were analyzed using receiver operating characteristic curve analysis. The results showed that the expression level of syncytin-1 in the serum of patients with newly diagnosed HCC was significantly higher than that in the normal control group (P < 0.0001). Using pathological diagnosis as the gold standard, the sensitivity and specificity of syncytin-1 for the auxiliary diagnosis of HCC were 91.3% and 75.5%, respectively, which were significantly higher than those of alpha-fetoprotein (P < 0.0001). The relative expression level of serum exosomal syncytin-1 was significantly associated with lymph node metastasis, degree of differentiation, and CNLC staging of HCC patients (P < 0.05). In conclusion, syncytin-1 in serum exosomes has high sensitivity and specificity for diagnosing HCC and can serve as a novel tumor marker for early screening, detection, and staging of HCC.

Clinical utility of electrocardiographic voltage parameters for the diagnosis of hypertrophic cardiomyopathy

BackgroundWhile electrocardiographic parameters of hypertensive left ventricular hypertrophy (H-LVH) are well known, limited data are available regarding hypertrophic cardiomyopathy (HCM). This study was to assess the diagnostic value of electrocardiographic voltage parameters in HCM.MethodsIncluded patients with HCM treated between March 2015 and May 2023. Voltage parameters (S-L, Cornell, Cornell product, Lewis, Peguero, and modified Cornell voltages) and echocardiography were evaluated. Receiver operating characteristic (ROC) curves were used to assess the diagnostic value of electrocardiogram in HCM. The multiple linear regression was conducted to analyze the correlation between electrocardiogram indicators and cardiac diastolic function.ResultsThe highest sensitivity for HCM was Peguero voltage (70.4%; 88.6% specificity). The Peguero voltage had high sensitivity in male (63.8%) and female patients (84.9%), those aged <65 years (71.6%) and ≥65 years (68.3%), with non-apical HCM (AHCM) (76.7%), obstructive HCM (82.1%), and non-obstructive HCM (66.9%). The sensitivity of the S-L voltage was high in AHCM (72.2%). The sensitivity for HCM reached 88.7% when the S-L and Peguero voltages were combined. The modified Cornell index had the highest area under the curve (0.88, 95% CI: 0.84–0.91), and its optimal cutoff value was 2.05 mV in males (77.6% sensitivity and 74% specificity) and 1.935 mV in females (90.6% sensitivity and 91.4% specificity). Peguero voltage (beta = 0.154, P = 0.034) and SD (beta = 0.223 P = 0.004) were independently correlated with E/e’, an index of left ventricular diastolic function.ConclusionThe Peguero voltage had high sensitivity and specificity for detecting the presence of HCM. It was positively correlated with E/e’ in patients with HCM. For AHCM, the S-L voltage was more advantageous. Combining the S-L voltage with the Peguero voltage further improves the sensitivity for HCM and thus could be used to improve the screening of HCM in clinical practice. The SD and modified Cornell voltage also had good diagnostic performance, especially in females.

High-Volume Production of Repeatable High Enhancement SERS Substrates Using Solid-State Superionic Stamping

Abstract Surface-enhanced Raman spectroscopy (SERS) is emerging as a powerful tool for detecting and identifying chemical and biological substances because of its high sensitivity, specificity, speed, and label-free detection. For SERS substrates to be effective in sensing applications, they must exhibit reproducible and robust high signal enhancement and cost-effective scalability. This article introduces a highly sensitive, large-area silver SERS substrate patterned with a uniform array of 3D retroreflecting inverted pyramids and develops a manufacturing pathway for it, using a novel and facile electrochemical imprinting process called solid-state superionic stamping (S4). Substrates, approximately 4 mm2 in area, are produced and tested with 1,2-bis(4-pyridyl) ethylene (BPE). Uniformly high and reproducible spatially averaged enhancement factor (EF), typically around a value of 2 × 107 with a relative standard deviation of 6.7% and a high batch-to-batch repeatability with a relative standard deviation of 10.5% between batches were observed. Passivating a substrate's surface with atomically thin layers of alumina, deposited using atomic layer deposition (ALD) was effective in maintaining the EF constant over a 60-day period, albeit with a trade-off between its EF and its lifespan. S4 has the potential to make substrates with EF consistently greater than 107 available at a cost of $1 to $2 per substrate, allowing SERS to be adopted across a wide spectrum of high-volume applications, including security, food safety, medical diagnostics, and chem-bio analysis.

Automated classification of mixed populations of Aedes aegypti and Culex quinquefasciatus mosquitoes under field conditions

BackgroundThe recent rise in the transmission of mosquito-borne diseases such as dengue virus (DENV), Zika (ZIKV), chikungunya (CHIKV), Oropouche (OROV), and West Nile (WNV) is a major concern for public health managers worldwide. Emerging technologies for automated remote mosquito classification can be supplemented to improve surveillance systems and provide valuable information regarding mosquito vector catches in real time.MethodsWe coupled an optical sensor to the entrance of a standard mosquito suction trap (BG-Mosquitaire) to record 9151 insect flights in two Brazilian cities: Rio de Janeiro and Brasilia. The traps and sensors remained in the field for approximately 1 year. A total of 1383 mosquito flights were recorded from the target species: Aedes aegypti and Culex quinquefasciatus. Mosquito classification was based on previous models developed and trained using European populations of Aedes albopictus and Culex pipiens.ResultsThe VECTRACK sensor was able to discriminate the target mosquitoes (Aedes and Culex genera) from non-target insects with an accuracy of 99.8%. Considering only mosquito vectors, the classification between Aedes and Culex achieved an accuracy of 93.7%. The sex classification worked better for Cx. quinquefasciatus (accuracy: 95%; specificity: 95.3%) than for Ae. aegypti (accuracy: 92.1%; specificity: 88.4%).ConclusionsThe data reported herein show high accuracy, sensitivity, specificity and precision of an automated optical sensor in classifying target mosquito species, genus and sex. Similar results were obtained in two different Brazilian cities, suggesting high reliability of our findings. Surprisingly, the model developed for European populations of Ae. albopictus worked well for Brazilian Ae. aegypti populations, and the model developed and trained for Cx. pipiens was able to classify Brazilian Cx. quinquefasciatus populations. Our findings suggest this optical sensor can be integrated into mosquito surveillance methods and generate accurate automatic real-time monitoring of medically relevant mosquito species.Graphical

Laminar specificity and coverage of viral-mediated gene expression restricted to GABAergic interneurons and their parvalbumin subclass in marmoset primary visual cortex

In the mammalian neocortex, inhibition is important for dynamically balancing excitation and shaping the response properties of cells and circuits. The various computational functions of inhibition are thought to be mediated by different inhibitory neuron types, of which a large diversity exists in several species. Current understanding of the function and connectivity of distinct inhibitory neuron types has mainly derived from studies in transgenic mice. However, it is unknown whether knowledge gained from mouse studies applies to the non-human primate, the model system closest to humans. The lack of viral tools to selectively access inhibitory neuron types has been a major impediment to studying their function in the primate. Here, we have thoroughly validated and characterized several recently developed viral vectors designed to restrict transgene expression to GABAergic cells or their parvalbumin (PV) subtype, and identified two types that show high specificity and efficiency in marmoset V1. We show that in marmoset V1, AAV-h56D induces transgene expression in GABAergic cells with up to 91–94% specificity and 79% efficiency, but this depends on viral serotype and cortical layer. AAV-PHP.eB-S5E2 induces transgene expression in PV cells across all cortical layers with up to 98% specificity and 86–90% efficiency, depending on layer. Thus, these viral vectors are promising tools for studying GABA and PV cell function and connectivity in the primate cortex.

Laminar specificity and coverage of viral-mediated gene expression restricted to GABAergic interneurons and their parvalbumin subclass in marmoset primary visual cortex

In the mammalian neocortex, inhibition is important for dynamically balancing excitation and shaping the response properties of cells and circuits. The various computational functions of inhibition are thought to be mediated by different inhibitory neuron types, of which a large diversity exists in several species. Current understanding of the function and connectivity of distinct inhibitory neuron types has mainly derived from studies in transgenic mice. However, it is unknown whether knowledge gained from mouse studies applies to the non-human primate, the model system closest to humans. The lack of viral tools to selectively access inhibitory neuron types has been a major impediment to studying their function in the primate. Here, we have thoroughly validated and characterized several recently developed viral vectors designed to restrict transgene expression to GABAergic cells or their parvalbumin (PV) subtype, and identified two types that show high specificity and efficiency in marmoset V1. We show that in marmoset V1, AAV-h56D induces transgene expression in GABAergic cells with up to 91–94% specificity and 79% efficiency, but this depends on viral serotype and cortical layer. AAV-PHP.eB-S5E2 induces transgene expression in PV cells across all cortical layers with up to 98% specificity and 86–90% efficiency, depending on layer. Thus, these viral vectors are promising tools for studying GABA and PV cell function and connectivity in the primate cortex.

Identification and Validation of Oxidative Stress-Related Biomarkers for Bronchopulmonary Dysplasia.

The objective of this study was to identify and characterize oxidative stress (OS)-related biomarkers in bronchopulmonary dysplasia (BPD) through a combination of bioinformatics analyses and wet experiments. The study utilized the Gene Expression Omnibus database to obtain the mRNA expression profile dataset GSE32472. Differential expression analysis and functional enrichment analysis were employed to investigate the role of OS-related genes in BPD. Gene Ontology Function Enrichment Analysis and Gene Set Enrichment Analysis were conducted to understand the mechanisms behind the signature. Protein-protein interaction analysis to identify hub genes in BPD, and predictions were made for microRNAs (miRNAs), transcription factors (TFs), and potential medications targeting these genes. CIBERSORT was utilized to investigate the correlation between hub genes and the infiltration of immune cells. Hub genes were ultimately determined and confirmed using expression analysis, correlation analysis, receiver operating characteristic (ROC) analysis, and quantitative real-time PCR (qRT-PCR). A novel OS-related gene signature (ARG1, CSF3R, IL1R1, IL1R2, MMP9, RETN, S100A12, and SOCS3) was constructed for the prediction of BPD. We identified 18 miRNAs, 14 TFs, and 30 potential medications targeting these genes. ROC analysis further validated that these genes could diagnose BPD with high specificity and sensitivity. The qRT-PCR revealed that IL1R1 and ARG1 were highly expressed in the lung tissue of the model group, while the expressions of RETN, SOCS3, IL1R2, and MMP9 were decreased. This study demonstrated that ARG1, CSF3R, IL1R1, IL1R2, MMP9, RETN, S100A12, and SOCS3 may serve as potential diagnostic biomarkers in BPD. Furthermore, a significant association between IL1R1 and the pathogenesis of BPD is observed.

Diagnostic Accuracy of the Second Edition of the Paris System for Reporting High-Grade Urothelial Carcinoma in Urinary Cytology.

Background Urinary cytology, a non-invasive screening tool, is essential for detecting high-grade urothelial neoplasms. The Paris System (TPS) standardizes reporting practices to improve diagnostic accuracy. TPS 2.0, introduced in 2022, categorizes samples into six diagnostic groups, emphasizing high-grade urothelial carcinoma (HGUC). Materials and Methods This retrospective study analyzed urine cytology samples from June 2023 to May 2024, correlating with histopathology when available. Samples were classified under TPS 2.0 categories, and statistical metrics including sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated for three groups based on malignancy criteria. Results Out of 180 samples, the distribution was: ND (3.9%), NHGUC (65.6%), AUC (10%), SHGUC (11.7%), and HGUC (8.9%). Histopathological correlation was available for 30.6% of cases. ROM values were: ND (33.3%), NHGUC (29.4%), AUC (66.7%), SHGUC (94.1%), and HGUC (100%). Group A showed the highest sensitivity (86.49%) and diagnostic accuracy (84.62%), while Group C had 100% specificity and PPV. Conclusion The study confirms TPS 2.0's efficacy in improving diagnostic accuracy for HGUC, with high specificity and sensitivity. Compared to TPS 1.0, TPS 2.0 offers clearer diagnostic criteria, enhancing clinical decision-making and patient outcomes. The findings support the continued use of TPS 2.0 in clinical practice, ensuring reliable identification of high-grade urothelial carcinomas.

Intratumoral calcification on CT assists in distinguishing benign and malignant upper abdomen neoplasm.

The significance of calcification and microcalcification in diagnosing malignant tumors is well established, but their role in the upper abdomen is less explored in routine radiology. To assess the effectiveness of computed tomography (CT) imaging in detecting intratumoral calcification within upper abdominal tumors. This study retrospectively enrolled patients with upper abdominal tumors featuring intratumoral calcifications who underwent plain and contrast-enhanced CT scans between January 2016 and December 2019. We examined the imaging characteristics of calcifications, including location, edges, shape, CT values, and association with necrosis. The diagnostic utility of calcification for distinguishing benign and malignant tumors was assessed using receiver operating characteristic curves. Univariate and multivariate logistic regression analyses were conducted to identify independent predictive factors for the diagnosis of malignancy characterized by intratumoral calcification. This study included 153 patients (median age 49 ± 21years; 83 men) with pathologically confirmed tumors of the upper abdomen (including liver, pancreas, and gastrointestinal tract) with intratumoral calcifications. Significant differences in CT values between benign and malignant tumors were observed (P < .001), with high diagnostic accuracy of calcification in CT imaging (receiver operating characteristic area = 0.884, sensitivity = 0.815, specificity = 0.976). The characteristics of calcification, including its edge and shape, were significantly correlated with tumor differentiation (P < .01). Multivariate logistic regression analysis revealed that the presence of adjacent necrosis around intracalcification is an independent predictor of malignancy (odds ratio = 5.48; 95% confidence interval: 1.55, 19.41; P = .008). Intratumoral calcification in CT imaging is a key marker for distinguishing between benign and malignant epigastric tumors, offering high specificity. Key message • What is already known on this topic - Intratumoral calcification, as a highly sensitive radiological marker, has shown potential in differentiating between benign and malignant tumors in thyroid and breast cancers. However, its discriminatory role in upper abdominal tumors is often overlooked. Therefore, assessing the diagnostic accuracy of intratumoral calcification on CT scans is crucial for improving diagnostic efficiency and avoiding unnecessary examinations. • What this study adds - Intratumoral calcification on CT exhibits high specificity in differentiating between benign and malignant upper abdominal tumors, providing a simple and reliable criterion for improving diagnostic accuracy. • How this study might affect research, practice or policy - This study highlights the significance of intratumoral calcification characteristics observed on CT in determining whether upper abdominal tumors are benign or malignant. The findings could pave the way for the development of a CT-based calcification scoring system, which would facilitate rapid and accurate diagnostics in clinical practice, thereby optimizing treatment strategies and enhancing patient prognosis.

Functionalized PLGA-Based Nanoparticles with Anti-HSV-2 Human Monoclonal Antibody: A Proof of Concept for Early Diagnosis and Targeted Therapy

Background: Functionalized nanoparticles (NPs) represent a cutting edge in innovative clinical approaches, allowing for the delivery of selected compounds with higher specificity in a wider time frame. They also hold promise for novel theranostic applications that integrate both diagnostic and therapeutic functions. Pathogens are continuously evolving to try to escape the strategies designed to treat them. Objectives: In this work, we describe the development of a biotechnological device, Nano-Immuno-Probes (NIPs), for early detection and infections treatment. Human Herpes Simplex Virus 2 was chosen as model pathogen. Methods: NIPs consist of PLGA-PEG-Sulfone polymeric NPs conjugated to recombinant Fab antibody fragments targeting the viral glycoprotein G2. NIPs synthesis involved multiple steps and was validated through several techniques. Results: DLS analysis indicated an expected size increase with a good polydispersity index. Z-average and z-potential values were measured for PLGA-PEG-Bis-Sulfone NPs (86.6 ± 10.9 nm; –0.7 ± 0.3 mV) and NIPs (151 ± 10.4 nm; −5.1 ± 1.9 mV). SPR assays confirmed NIPs’ specificity for the glycoprotein G2, with an apparent KD of 1.03 ± 0.61 µM. NIPs exhibited no cytotoxic effects on VERO cells at 24 and 48 h. Conclusions: This in vitro study showed that NIPs effectively target HSV-2, suggesting the potential use of these nanodevices to deliver both contrast agents as well as therapeutic compounds.

Lateral pelvic lymph node positivity (LPLNP) score: predictive clinic-radiological model of lateral pelvic lymph node involvement in rectal cancer patients

PurposeThe population in Western countries differs significantly from that in Eastern countries, and the prevalence of lateral pelvic lymph node (LPLN) involvement in Western populations remains largely unknown due to the limited application of LPLN dissection (LPLND). This discrepancy is primarily attributed to the higher body mass index commonly observed in Western populations, which increases the risk of intraoperative complications. Consequently, the aim of this study is to describe a specific Western clinico-radiological selection tool for LPLND, namely, the lateral pelvic lymph node positivity (LPLNP) score.MethodsThis retrospective single center study was designed to elaborate the LPLNP score, which was further tested on a prospective cohort of patients. Clinical and MRI factors associated with LPLN involvement were identified, and logistic regression was used to establish the LPLNP score.ResultsIn the retrospective series, 120 patients underwent lateral pelvic lymph node dissection. After stepwise logistic regression, five parameters were ultimately included in the LPLNP score. When tested on 66 prospectively selected patients, 40 with an LPLNP score > 0.23 (corresponding to the highest sensitivity and specificity) underwent LPLND: 22 patients (55%) had pathologically confirmed positive LPLN. The negative predictive value of the LPLNP score was 96%, with a sensitivity of 95.7% and a specificity of 58.1%.ConclusionThe LPLNP score was developed based on the largest group of Western patients with locally advanced rectal cancer. This scoring system demonstrated high sensitivity and specificity during validation on the prospective series, correctly identifying LPLN involvement in 55% of cases.