An automated framework for pediatric hip surveillance and severity assessment using radiographs.

PURPOSE: Hip dysplasia is the second most common orthopedic condition in children with cerebral palsy (CP) and may result in disability and pain. The migration percentage (MP) is a widely used metric in hip surveillance, calculated based on an anterior-posterior pelvis radiograph. However, manual quantification of MP values using hip X-ray scans in current standard practice has challenges including being time-intensive, requiring expert knowledge, and not considering human bias. The purpose of this study is to develop a machine learning algorithm to automatically quantify MP values using a hip X-ray scan, and hence provide an assessment for severity, which then can be used for surveillance, treatment planning, and management. METHODS: X-ray scans from 210 patients were curated, pre-processed, and manually annotated at our clinical center. Several machine learning models were trained using pre-trained weights from Inception ResNet-V2, VGG-16, and VGG-19, with different strategies (pre-processing, with and without region of interest (ROI) detection, with and without data augmentation) to find an optimal model for automatic hip landmarking. The predicted landmarks were then used by our geometric algorithm to quantify the MP value for the input hip X-ray scan. RESULTS: The pre-trained VGG-19 model, fine-tuned with additional custom layers, outputted the lowest mean squared error values for both train and test data, when ROI cropped images were used along with data augmentation for model training. The MP value calculated by the algorithm was compared to manual ground truth labels from our orthopedic fellows using the hip screen application for benchmarking. CONCLUSION: The results showed the feasibility of the machine learning model in automatic hip landmark detection for reliably quantifying MP value from hip X-ray scans. The algorithm could be used as an accurate and reliable tool in orthopedic care for diagnosing, severity assessment, and hence treatment and surgical planning for hip displacement.

Farnesoid X receptor activation by bile acids suppresses lipid peroxidation and ferroptosis.

Ferroptosis is a regulated cell death modality that occurs upon iron-dependent lipid peroxidation. Recent research has identified many regulators that induce or inhibit ferroptosis; yet, many regulatory processes and networks remain to be elucidated. In this study, we performed a chemical genetics screen using small molecules with known mode of action and identified two agonists of the nuclear receptor Farnesoid X Receptor (FXR) that suppress ferroptosis, but not apoptosis or necroptosis. We demonstrate that in liver cells with high FXR levels, knockout or inhibition of FXR sensitized cells to ferroptotic cell death, whereas activation of FXR by bile acids inhibited ferroptosis. Furthermore, FXR inhibited ferroptosis in ex vivo mouse hepatocytes and human hepatocytes differentiated from induced pluripotent stem cells. Activation of FXR significantly reduced lipid peroxidation by upregulating the ferroptosis gatekeepers GPX4, FSP1, PPARα, SCD1, and ACSL3. Together, we report that FXR coordinates the expression of ferroptosis-inhibitory regulators to reduce lipid peroxidation, thereby acting as a guardian of ferroptosis.

The changing characteristics of a cohort of children and adolescents living with HIV at antiretroviral therapy initiation in Asia.

Despite improvements in HIV testing and earlier antiretroviral therapy (ART) initiation in children living with HIV through the years, a considerable proportion start treatment with advanced disease. We studied characteristics of children and adolescents living with HIV and their level of immunodeficiency at ART initiation using data from a multi-country Asian cohort. We included children and adolescents who were ART-naïve and <18 years of age at ART initiation from 2011 to 2020 at 17 HIV clinics in six countries. Incidence rates of opportunistic infections (OIs) in the first two years of triple-drug ART (≥3 antiretrovirals) was also reported. Competing risk regression analysis was performed to identify factors associated with first occurrence of OI. In 2,027 children and adolescents (54% males), median age at ART initiation increased from 4.5 years in 2011-2013 to 6.7 in 2017-2020, median CD4 count doubled from 237 cells/µl to 466 cells/µl, and proportion of children who initiated ART as severely immunodeficient decreased from 70% to 45%. During follow-up, 275 (14%) children who received triple-drug ART as first treatment and had at least one clinic visit, developed at least one OI in the first two years of treatment (9.40 per 100 person-years). The incidence rate of any first OI declined from 12.52 to 7.58 per 100 person-years during 2011-2013 and 2017-2020. Lower hazard of OIs were found in those with age at first ART 2-14 years, current CD4 ≥200 cells/µl, and receiving ART between 2017 and 2020. The analysis demonstrated increasing number of children and adolescents starting ART with high CD4 count at ART start. The rate of first OI markedly decreased in children who started ART in more recent years. There remains a clear need for improvement in HIV control strategies in children, by promoting earlier diagnosis and timely treatment.

Effect of Pharmacist-Led Interventions on Medication Adherence among Vietnamese Patients with Asthma: A Randomized Controlled Trial.

BACKGROUND: Medication adherence in asthmatic patients enhances the effectiveness of treatments, but some studies in low and middle-income countries still show some limitations. Our study aimed to determine if pharmacist-led interventions could increase medication adherence, improve treatment effectiveness, and relieve symptom severity in outpatients with asthma. METHODS: We conducted a randomized, controlled trial on 247 asthmatic outpatients (aged ≥ 16) with a 1:1 ratio randomization at the hospitalization time and repeated after 1-month discharge. The primary outcome was to detect the difference in medication adherence between groups. Adherence was assessed by the general medication adherence scale (GMAS). Data collected by questionnaire was coded and entered into SPSS_20 for statistical analysis; Results: 247 patients (123 intervention, 124 control) were enrolled (61.1% male). After intervention, the adherence rate was higher among the intervention group than the control group (94.3% vs. 82.8%, p = 0.001). Patient behavior and knowledge were enhanced in the intervention group (p < 0.05). Asthma symptoms were relieved in the intervention group (p = 0.014). Pharmacist-led interventions on adherence rate were higher with OR: 3.550, 95% CI: 1.378-9.143, p = 0.009. CONCLUSIONS: pharmaceutical intervention could improve medication adherence, treatment efficacy, and the outcome should not be taken for granted; further research should be carried out in this regard.

Digital Holographic Microscopy to Assess Cell Behavior.

Digital holographic microscopy is an imaging technique particularly well suited to the study of living cells in culture, as no labeling is required and computed phase maps produce high contrast, quantitative pixel information. A full experiment involves instrument calibration, cell culture quality checks, selection and setup of imaging chambers, a sampling plan, image acquisition, phase and amplitude map reconstruction, and parameter map post-processing to extract information about cell morphology and/or motility. Each step is described below, focusing on results from imaging four human cell lines. Several post-processing approaches are detailed, with an aim of tracking individual cells and dynamics of cell populations.

Generalized Fringe-to-Phase Framework for Single-Shot 3D Reconstruction Integrating Structured Light with Deep Learning.

Three-dimensional (3D) shape acquisition of objects from a single-shot image has been highly demanded by numerous applications in many fields, such as medical imaging, robotic navigation, virtual reality, and product in-line inspection. This paper presents a robust 3D shape reconstruction approach integrating a structured-light technique with a deep learning-based artificial neural network. The proposed approach employs a single-input dual-output network capable of transforming a single structured-light image into two intermediate outputs of multiple phase-shifted fringe patterns and a coarse phase map, through which the unwrapped true phase distributions containing the depth information of the imaging target can be accurately determined for subsequent 3D reconstruction process. A conventional fringe projection technique is employed to prepare the ground-truth training labels, and part of its classic algorithm is adopted to preserve the accuracy of the 3D reconstruction. Numerous experiments have been conducted to assess the proposed technique, and its robustness makes it a promising and much-needed tool for scientific research and engineering applications.

Disclosure of HIV status and associated clinical outcomes of children and adolescents living with HIV in Asia.

Disclosure of HIV status is an important part of pediatric care. We studied disclosure and clinical outcomes in a multi-country Asian cohort of children and adolescents with HIV. Those 6-19 years of age who initiated combination antiretroviral therapy (cART) between 2008 and 2018, and who had at least one follow-up clinic visit were included. Data up to December 2019 were analyzed. Cox and competing risk regression analyses were used to assess the effect of disclosure on disease progression (WHO clinical stage 3 or 4), loss to follow-up (LTFU; > 12 months), and death. Of 1913 children and adolescents (48% female; median [IQR] age 11.5 [9.2-14.7] years at last clinic visit), 795 (42%) were disclosed to about their HIV status at a median age of 12.9 years (IQR: 11.8-14.1). During follow-up, 207 (11%) experienced disease progression, 75 (3.9%) were LTFU, and 59 (3.1%) died. There were lower hazards of disease progression (adjusted hazard ratio [aHR] 0.43 [0.28-0.66]) and death (aHR 0.36 [0.17-0.79]) for those disclosed to compared with those who were not. Disclosure and its appropriate implementation should be promoted in pediatric HIV clinics in resource-limited settings.

Pregnancy and birth outcomes among young women living with perinatally acquired HIV in Thailand and Vietnam.

We conducted a retrospective cohort study of pregnancy and infant outcomes in 670 adolescents and young adult women with perinatally acquired HIV (AYAPHIV), aged 15-24 years, in Thailand and Vietnam. Between January 2013 and December 2018, there were 52 pregnancies, for an incidence of 2.49 (95% CI 1.90-3.27) per 100 person-years. The median age at pregnancy was 17.7 years (IQR 16.8-18.9). Pregnant AYAPHIV had been on cART for a lifetime median of 9.8 years (IQR 7.3-12.4). At the time of conception, the median CD4 was 521 cells/mm3 (IQR 213-760), and 76% had HIV RNA ≤400 copies/ml. Of the 51 pregnancies with available outcomes, 90% resulted in live singleton births at a median gestational age of 38 weeks (IQR 37-39); 77% of mothers (n = 27/35) had HIV RNA ≤400 copies/ml at delivery. Among infants with available data, 50% (n = 21/42) were male and 29% (n = 12/42) were reported to be low birthweight (<2,500gm); none (n = 0/41) were breastfed. One infant was diagnosed with HIV. Our findings emphasize that efforts to strengthen reproductive health education, including contraception, pregnancy-related psychosocial support services, and prevention of vertical HIV transmission interventions, in our region are needed for adolescents with perinatally acquired HIV as they transition to young adults.

Versatile role of Pseudomonas fuscovaginae cyclic lipopeptides in plant and microbial interactions.

Pseudomonas fuscovaginae is the most prominent bacterial sheath rot pathogen, causing sheath brown rot disease in rice. This disease occurs worldwide and it is characterized by typical necrotic lesions on the sheath, as well as a reduction in the number of emitted panicles and filled grains. P. fuscovaginae has been shown to produce syringotoxin and fuscopeptin cyclic lipopeptides (CLPs), which have been linked to pathogenicity. In this study, we investigated the role of P. fuscovaginae UPB0736 CLPs in plant pathogenicity, antifungal activity and swarming motility. To do so, we sequenced the strain to obtain a single-contig genome and we constructed deletion mutants in the biosynthetic gene clusters responsible for the synthesis of CLPs. We show that UPB0736 produces a third CLP of 13 amino acids, now named asplenin, and we link this CLP with the swarming activity of the strain. We could then show that syringotoxin is particularly active against Rhizoctonia solani in vitro. By testing the mutants in planta we investigated the role of both fuscopeptin and syringotoxin in causing sheath rot lesions. We proved that the presence of these two CLPs considerably affected the number of emitted panicles, although their number was still significantly affected in the mutants deficient in both fuscopeptin and syringotoxin. These results reveal the importance of CLPs in P. fuscovaginae pathogenicity, but also suggest that other pathogenicity factors may be involved.

Design, synthesis, and characterization of novel fluorogenic substrates of the proprotein convertases furin, PC1/3, PC2, PC5/6, and PC7.

Proprotein convertases (PCs) are involved in the pathogenesis of various diseases, making them promising drug targets. Most assays for PCs have been performed with few standard substrates, regardless of differences in cleavage efficiencies. Derived from studies on substrate-analogue inhibitors, 11 novel substrates were synthesized and characterized with five PCs. H-Arg-Arg-Tle-Lys-Arg-AMC is the most efficiently cleaved furin substrate based on its kcat/KM value. Due to its higher kcat value, acetyl-Arg-Arg-Tle-Arg-Arg-AMC was selected for further measurements to demonstrate the benefit of this improved substrate. Compared to our standard conditions, its use allowed a 10-fold reduction of the furin concentration, which enabled Ki value determinations of previously described tight-binding inhibitors under classical conditions. Under these circumstances, a slow-binding behavior was observed for the first time with inhibitor MI-1148. In addition to furin, four additional PCs were used to characterize these substrates. The most efficiently cleaved PC1/3 substrate was acetyl-Arg-Arg-Arg-Tle-Lys-Arg-AMC. The highest kcat/KM values for PC2 and PC7 were found for the N-terminally unprotected analogue of this substrate, although other substrates possess higher kcat values. The highest efficiency for PC5/6A was observed for the substrate acetyl-Arg-Arg-Tle-Lys-Arg-AMC. In summary, we have identified new substrates for furin, PC1/3, PC2, and PC7 suitable for improved enzyme-kinetic measurements.

Childhood encephalitis in the Greater Mekong region (the SouthEast Asia Encephalitis Project): a multicentre prospective study.

BACKGROUND: Encephalitis is a worldwide public health issue, with a substantially high burden among children in southeast Asia. We aimed to determine the causes of encephalitis in children admitted to hospitals across the Greater Mekong region by implementing a comprehensive state-of-the-art diagnostic procedure harmonised across all centres, and identifying clinical characteristics related to patients' conditions. METHODS: In this multicentre, observational, prospective study of childhood encephalitis, four referral hospitals in Cambodia, Vietnam, Laos, and Myanmar recruited children (aged 28 days to 16 years) who presented with altered mental status lasting more than 24 h and two of the following minor criteria: fever (within the 72 h before or after presentation), one or more generalised or partial seizures (excluding febrile seizures), a new-onset focal neurological deficit, cerebrospinal fluid (CSF) white blood cell count of 5 per mL or higher, or brain imaging (CT or MRI) suggestive of lesions of encephalitis. Comprehensive diagnostic procedures were harmonised across all centres, with first-line testing was done on samples taken at inclusion and results delivered within 24 h of inclusion for main treatable causes of disease and second-line testing was done thereafter for mostly non-treatable causes. An independent expert medical panel reviewed the charts and attribution of causes of all the included children. Using multivariate analyses, we assessed risk factors associated with unfavourable outcomes (ie, severe neurological sequelae and death) at discharge using data from baseline and day 2 after inclusion. This study is registered with ClinicalTrials.gov, NCT04089436, and is now complete. FINDINGS: Between July 28, 2014, and Dec 31, 2017, 664 children with encephalitis were enrolled. Median age was 4·3 years (1·8-8·8), 295 (44%) children were female, and 369 (56%) were male. A confirmed or probable cause of encephalitis was identified in 425 (64%) patients: 216 (33%) of 664 cases were due to Japanese encephalitis virus, 27 (4%) were due to dengue virus, 26 (4%) were due to influenza virus, 24 (4%) were due to herpes simplex virus 1, 18 (3%) were due to Mycobacterium tuberculosis, 17 (3%) were due to Streptococcus pneumoniae, 17 (3%) were due to enterovirus A71, 74 (9%) were due to other pathogens, and six (1%) were due to autoimmune encephalitis. Diagnosis was made within 24 h of admission to hospital for 83 (13%) of 664 children. 119 (18%) children had treatable conditions and 276 (42%) had conditions that could have been preventable by vaccination. At time of discharge, 153 (23%) of 664 children had severe neurological sequelae and 83 (13%) had died. In multivariate analyses, risk factors for unfavourable outcome were diagnosis of M tuberculosis infection upon admission (odds ratio 3·23 [95% CI 1·04-10·03]), coma on day 2 (2·90 [1·78-4·72]), supplementary oxygen requirement (1·89 [1·25-2·86]), and more than 1 week duration between symptom onset and admission to hospital (3·03 [1·68-5·48]). At 1 year after inclusion, of 432 children who were discharged alive from hospital with follow-up data, 24 (5%) had died, 129 (30%) had neurological sequelae, and 279 (65%) had completely recovered. INTERPRETATION: In southeast Asia, most causes of childhood encephalitis are either preventable or treatable, with Japanese encephalitis virus being the most common cause. We provide crucial information that could guide public health policy to improve diagnostic, vaccination, and early therapeutic guidelines on childhood encephalitis in the Greater Mekong region. FUNDING: Institut Pasteur, Institut Pasteur International Network, Fondation Merieux, Aviesan Sud, INSERM, Wellcome Trust, Institut de Recherche pour le Développement (IRD), and Fondation Total.

Dual-modality digital holographic and polarization microscope to quantify phase and birefringence signals in biospecimens with a complex microstructure.

Optical phase and birefringence signals occur in cells and thin, semi-transparent biomaterials. A dual-modality quantitative phase and polarization microscope was designed to study the interaction of cells with extracellular matrix networks and to relate optical pathlength and birefringence signals within structurally anisotropic biomaterial constructs. The design was based on an existing, custom-built digital holographic microscope, to which was added a polarization microscope utilizing liquid crystal variable retarders. Phase and birefringence channels were calibrated, and data was acquired sequentially from cell-seeded collagen hydrogels and electrofabricated chitosan membranes. Computed phase height and retardance from standard targets were accurate within 99.7% and 99.8%, respectively. Phase height and retardance channel background standard deviations were 35 nm and 0.6 nm, respectively. Human fibroblasts, visible in the phase channel, aligned with collagen network microstructure, with retardance and azimuth visible in the polarization channel. Electrofabricated chitosan membranes formed in 40 µm tall microfluidic channels possessed optical retardance ranging from 7 to 11 nm, and phase height from 37 to 39 µm. These results demonstrate co-registered dual-channel acquisition of phase and birefringence parameter maps from microstructurally-complex biospecimens using a novel imaging system combining digital holographic microscopy with voltage-controlled polarization microscopy.

Impact of antimicrobial stewardship intervention in clean and clean-contaminated surgical procedures at a Vietnamese national hospital.

OBJECTIVES: We investigated the characteristics of prophylactic antimicrobial use in clean and clean-contaminated surgical procedures and assessed the efficacy of a prophylactic antimicrobial stewardship intervention at Thong Nhat Hospital, Ho Chi Minh City, Vietnam. METHODS: A cross-sectional study was conducted on 354 patients who underwent either clean or clean-contaminated surgical procedures at Thong Nhat Hospital. Eligible patients were classified with respect to three periods of intervention from 2017 to 2020. Data collection included surgical procedures, patient characteristics, and prophylactic antimicrobial usage. We determined the efficacy of antimicrobial stewardship intervention based on comparisons among the primary outcome (the appropriateness of prophylactic antimicrobials) and secondary outcomes (postoperative antimicrobial prophylaxis (AP) prolongation, length of postoperative hospital stay, and cost of antimicrobials). RESULTS: The mean age of patients in periods 1, 2, and 3 was 54.5 ± 16.6, 50.2 ± 16.5, and 52.8 ± 17.3 years, respectively, with an overall male/female ratio of 1.1/1. No significant differences were detected in basic patient characteristics during the three periods. Majority of the surgical procedures were clean (56%-59%) and scheduled (85%-86%). Prophylactic antimicrobial stewardship intervention enhanced AP appropriateness (by 12.7%, 12.7%, and 39.0% in periods 1, 2, and 3, respectively, p < 0.001), decreased postoperative prophylactic antimicrobial duration [3.0 (0-6), 1.5 (0-5), and 0.0 (0-1) days, respectively, p < 0.001], and reduced average antimicrobial expenses (p < 0.001). CONCLUSIONS: The prophylactic antimicrobial stewardship interventions introduced at Thong Nhat Hospital had several positive impacts on the appropriateness of prophylactic antimicrobial use and treatment costs.

Facile Fabrication of Novel NiFe2O4@Carbon Composites for Enhanced Adsorption of Emergent Antibiotics.

Water purification is becoming one of the most pertinent environmental issues throughout the world. Among common types of water pollution involving heavy metals, pharmaceutical drugs, textile dyes, personal care products, and other persistent organic pollutants, the pollution of antibiotic drugs is increasingly emerging due to their adverse effects on microorganisms, aquatic animals, and human health. Therefore, the treatment of such contaminants is very necessary to reduce the concentration of antibiotic pollutants to permissible levels prior to discharge. Herein, we report the use of NiFe2O4@C composites from a bimetallic-based metal-organic framework Ni-MIL-88B(Fe) for removal of ciprofloxacin (CFX) and tetracycline (TCC). The effect of production temperatures (600-900 °C), solution pH (2-10), NiFe2O4@C dose (0.05-0.2 g/L), concentration of antibiotics (10-60 mg/L), and uptake time (0-480 min) was investigated systematically. Response surface methodology and central composite design were applied for quadratic models to discover optimum conditions of antibiotic adsorption. With high coefficients of determination (R2 = 0.9640-0.9713), the proposed models were significant statistically. Under proposed optimum conditions, the adsorption capacity for CFX and TCC were found at 256.244, and 105.38 mg/g, respectively. Recyclability study was employed and found that NiFe2O4@C-900 could be reused for up to three cycles, offering the potential of this composite as a good adsorbent for removal of emergent antibiotics.

Alterations in children's sub-dominant gut microbiota by HIV infection and anti-retroviral therapy.

OBJECTIVE: We investigated the impact of human immunodeficiency virus (HIV) infection and anti-retroviral therapy (ART) on the gut microbiota of children. DESIGN: This cross-sectional study investigated the gut microbiota of children with and without HIV. METHODS: We collected fecal samples from 59 children with HIV (29 treated with ART [ART(+)] and 30 without ART [HIV(+)]) and 20 children without HIV [HIV(-)] in Vietnam. We performed quantitative RT-PCR to detect 14 representative intestinal bacteria targeting 16S/23S rRNA molecules. We also collected the blood samples for immunological analyses. RESULTS: In spearman's correlation analyses, no significant correlation between the number of dominant bacteria and age was found among children in the HIV(-) group. However, the number of sub-dominant bacteria, including Streptococcus, Enterococcus, and Enterobacteriaceae, positively correlated with age in the HIV(-) group, but not in the HIV(+) group. In the HIV(+) group, Clostridium coccoides group positively associated with the CD4+ cell count and its subsets. In the ART(+) group, Staphylococcus and C. perfringens positively correlated with CD4+ cells and their subsets and negatively with activated CD8+ cells. C. coccoides group and Bacteroides fragilis group were associated with regulatory T-cell counts. In multiple linear regression analyses, ART duration was independently associated with the number of C. perfringens, and Th17 cell count with the number of Staphylococcus in the ART(+) group. CONCLUSIONS: HIV infection and ART may influence sub-dominant gut bacteria, directly or indirectly, in association with immune status in children with HIV.

Bacillus Cyclic Lipopeptides Iturin and Fengycin Control Rice Blast Caused by Pyricularia oryzae in Potting and Acid Sulfate Soils by Direct Antagonism and Induced Systemic Resistance.

Rice monoculture in acid sulfate soils (ASSs) is affected by a wide range of abiotic and biotic constraints, including rice blast caused by Pyricularia oryzae. To progress towards a more sustainable agriculture, our research aimed to screen the biocontrol potential of indigenous Bacillus spp. against blast disease by triggering induced systemic resistance (ISR) via root application and direct antagonism. Strains belonging to the B. altitudinis and B. velezensis group could protect rice against blast disease by ISR. UPLC-MS and marker gene replacement methods were used to detect cyclic lipopeptide (CLiP) production and construct CLiPs deficient mutants of B. velezensis, respectively. Here we show that the CLiPs fengycin and iturin are both needed to elicit ISR against rice blast in potting soil and ASS conditions. The CLiPs surfactin, iturin and fengycin completely suppressed P. oryzae spore germination resulting in disease severity reduction when co-applied on rice leaves. In vitro microscopic assays revealed that iturin and fengycin inhibited the mycelial growth of the fungus P. oryzae, while surfactin had no effect. The capacity of indigenous Bacillus spp. to reduce rice blast by direct and indirect antagonism in ASS conditions provides an opportunity to explore their usage for rice blast control in the field.

The Basicity Makes the Difference: Improved Canavanine-Derived Inhibitors of the Proprotein Convertase Furin.

Furin activates numerous viral glycoproteins, and its inhibition prevents virus replication and spread. Through the replacement of arginine by the less basic canavanine, new inhibitors targeting furin in the trans-Golgi network were developed. These inhibitors exert potent antiviral activity in cell culture with much lower toxicity than arginine-derived analogues, most likely due to their reduced protonation in the blood circulation. Thus, despite its important physiological functions, furin might be a suitable antiviral drug target.

Keratin 19 maintains E-cadherin localization at the cell surface and stabilizes cell-cell adhesion of MCF7 cells.

A cytoskeletal protein keratin 19 (K19) is highly expressed in breast cancer but its effects on breast cancer cell mechanics are unclear. In MCF7 cells where K19 expression is ablated,we found that K19 is required to maintain rounded epithelial-like shape and tight cell-cell adhesion. A loss of K19 also lowered cell surface E-cadherin levels. Inhibiting internalization restored cell-cell adhesion of KRT19  knockout cells, suggesting that E-cadherin internalization contributed to defective adhesion. Ultimately, while K19 inhibited cell migration and invasion, it was required for cells to form colonies in suspension. Our results suggest that K19 stabilizes E-cadherin complexes at the cell membrane to maintain cell-cell adhesion which inhibits cell invasiveness but provides growth and survival advantages for circulating tumor cells.

Molecular genotypes of gag sequences in HIV-1 infected children treated with antiretroviral therapy in Vietnam.

BACKGROUND: Gag protein of human immunodeficiency virus (HIV) has been reported to play a crucial role in establishing infection, viral replication, and disease progression; thus, gag might be related to treatment response. The objective of this study was to investigate molecular genotypes of the gag gene, particularly the important functional binding domains in relation to treatment outcomes. METHODS: HIV-infected children enrolled and treated at Vietnam National Children's Hospital were recruited in the study. A total of 25 gag sequences were generated and used to construct phylogenetic trees and aligned with a reference sequence comparing 17 functional domains. RESULTS: We found that all patients in a treatment failure (TF) group belonged to one cluster of the phylogenetic tree. In addition, the rate of mutations was significantly higher in TF compared with a treatment success (TS) group, specifically the PIP2 recognition motif, and the nucleocapsid basic and zinc motif 2 domains [median and (interquartile range (IQR): 12.5 (6.25-12.5) versus 50 (25-50), p < 0.01; 0 (0-0) versus 0 (0-21.43), p = 0.03 and 0 (0-7.14) versus 7.14 (7.14-7.14), p = 0.04, respectively]. When analyzing gag sequences at different time points in seven patients, we did not observe a consistent mutation pattern related to treatment response. CONCLUSION: Gag mutations in certain domains might be associated with increased viral load; therefore, studying the molecular genotype of the gag gene might be beneficial in monitoring treatment response in HIV-infected children.

Virtual organelle self-coding for fluorescence imaging via adversarial learning.

SIGNIFICANCE: Our study introduces an application of deep learning to virtually generate fluorescence images to reduce the burdens of cost and time from considerable effort in sample preparation related to chemical fixation and staining. AIM: The objective of our work was to determine how successfully deep learning methods perform on fluorescence prediction that depends on structural and/or a functional relationship between input labels and output labels. APPROACH: We present a virtual-fluorescence-staining method based on deep neural networks (VirFluoNet) to transform co-registered images of cells into subcellular compartment-specific molecular fluorescence labels in the same field-of-view. An algorithm based on conditional generative adversarial networks was developed and trained on microscopy datasets from breast-cancer and bone-osteosarcoma cell lines: MDA-MB-231 and U2OS, respectively. Several established performance metrics-the mean absolute error (MAE), peak-signal-to-noise ratio (PSNR), and structural-similarity-index (SSIM)-as well as a novel performance metric, the tolerance level, were measured and compared for the same algorithm and input data. RESULTS: For the MDA-MB-231 cells, F-actin signal performed the fluorescent antibody staining of vinculin prediction better than phase-contrast as an input. For the U2OS cells, satisfactory metrics of performance were archieved in comparison with ground truth. MAE is <0.005, 0.017, 0.012; PSNR is >40 / 34 / 33 dB; and SSIM is >0.925 / 0.926 / 0.925 for 4',6-diamidino-2-phenylindole/hoechst, endoplasmic reticulum, and mitochondria prediction, respectively, from channels of nucleoli and cytoplasmic RNA, Golgi plasma membrane, and F-actin. CONCLUSIONS: These findings contribute to the understanding of the utility and limitations of deep learning image-regression to predict fluorescence microscopy datasets of biological cells. We infer that predicted image labels must have either a structural and/or a functional relationship to input labels. Furthermore, the approach introduced here holds promise for modeling the internal spatial relationships between organelles and biomolecules within living cells, leading to detection and quantification of alterations from a standard training dataset.