HyU: Hybrid Unmixing for longitudinal in vivo imaging of low signal-to-noise fluorescence.

The expansion of fluorescence bioimaging toward more complex systems and geometries requires analytical tools capable of spanning widely varying timescales and length scales, cleanly separating multiple fluorescent labels and distinguishing these labels from background autofluorescence. Here we meet these challenging objectives for multispectral fluorescence microscopy, combining hyperspectral phasors and linear unmixing to create Hybrid Unmixing (HyU). HyU is efficient and robust, capable of quantitative signal separation even at low illumination levels. In dynamic imaging of developing zebrafish embryos and in mouse tissue, HyU was able to cleanly and efficiently unmix multiple fluorescent labels, even in demanding volumetric timelapse imaging settings. HyU permits high dynamic range imaging, allowing simultaneous imaging of bright exogenous labels and dim endogenous labels. This enables coincident studies of tagged components, cellular behaviors and cellular metabolism within the same specimen, providing more accurate insights into the orchestrated complexity of biological systems.

FRaeppli: a multispectral imaging toolbox for cell tracing and dense tissue analysis in zebrafish.

Visualizing cell shapes and interactions of differentiating cells is instrumental for understanding organ development and repair. Across species, strategies for stochastic multicolour labelling have greatly facilitated in vivo cell tracking and mapping neuronal connectivity. Yet integrating multi-fluorophore information into the context of developing zebrafish tissues is challenging given their cytoplasmic localization and spectral incompatibility with common fluorescent markers. Inspired by Drosophila Raeppli, we developed FRaeppli (Fish-Raeppli) by expressing bright membrane- or nuclear-targeted fluorescent proteins for efficient cell shape analysis and tracking. High spatiotemporal activation flexibility is provided by the Gal4/UAS system together with Cre/lox and/or PhiC31 integrase. The distinct spectra of the FRaeppli fluorescent proteins allow simultaneous imaging with GFP and infrared subcellular reporters or tissue landmarks. We demonstrate the suitability of FRaeppli for live imaging of complex internal organs, such as the liver, and have tailored hyperspectral protocols for time-efficient acquisition. Combining FRaeppli with polarity markers revealed previously unknown canalicular topologies between differentiating hepatocytes, reminiscent of the mammalian liver, suggesting common developmental mechanisms. The multispectral FRaeppli toolbox thus enables the comprehensive analysis of intricate cellular morphologies, topologies and lineages at single-cell resolution in zebrafish.

Macrophage Epithelial Reprogramming Underlies Mycobacterial Granuloma Formation and Promotes Infection.

Mycobacterium tuberculosis infection in humans triggers formation of granulomas, which are tightly organized immune cell aggregates that are the central structure of tuberculosis. Infected and uninfected macrophages interdigitate, assuming an altered, flattened appearance. Although pathologists have described these changes for over a century, the molecular and cellular programs underlying this transition are unclear. Here, using the zebrafish-Mycobacterium marinum model, we found that mycobacterial granuloma formation is accompanied by macrophage induction of canonical epithelial molecules and structures. We identified fundamental macrophage reprogramming events that parallel E-cadherin-dependent mesenchymal-epithelial transitions. Macrophage-specific disruption of E-cadherin function resulted in disordered granuloma formation, enhanced immune cell access, decreased bacterial burden, and increased host survival, suggesting that the granuloma can also serve a bacteria-protective role. Granuloma macrophages in humans with tuberculosis were similarly transformed. Thus, during mycobacterial infection, granuloma macrophages are broadly reprogrammed by epithelial modules, and this reprogramming alters the trajectory of infection and the associated immune response.

A real-world cohort study of first-line afatinib in patients with EGFR-mutant advanced non-small cell lung cancer in Vietnam.

BACKGROUND: This study aimed to evaluate the efficacy and side effects of first-line afatinib treatment in a real-world setting in Vietnam. METHODS: This retrospective study was conducted across nine hospitals in Vietnam. Advanced epidermal growth factor receptor (EGFR)-mutant non-small cell lung cancer (NSCLC) patients who received afatinib as first-line therapy between April 2018 and June 2022 were included, and patient medical records were reviewed. Key outcomes were overall response rate (ORR), time-to-treatment failure (TTF), and tolerability. RESULTS: A total of 343 patients on first-line afatinib were eligible for the study. EGFR exon 19 deletion (Del19) alone was detected in 46.9% of patients, L858R mutation alone in 26.3%, and other uncommon EGFR mutations, including compound mutations, in 26.8%. Patients with brain metastases at baseline were 25.4%. Patients who received 40 mg, 30 mg, and 20 mg as starting doses of afatinib were 58.6%, 39.9%, and 1.5%, respectively. The ORR was 78.1% in the overall population, 82.6% in the Del19 mutation subgroup, 73.3% in the L858R mutation subgroup, and 75.0% in the uncommon mutation subgroup (p > 0.05). The univariate and multivariate analyses indicate that the ORR increased when the starting dose was 40 mg compared to starting doses below 40 mg (83.9% vs. 74.3%, p = 0.034). The median TTF (mTTF) was 16.7 months (CI 95%: 14.8-18.5) in all patients, with a median follow-up time of 26.2 months. The mTTF was longer in patients in the common EGFR mutation subgroup (Del19/L858R) than in those in the uncommon mutation subgroup (17.5 vs. 13.8 months, p = 0.045) and in those without versus with brain metastases at baseline (17.5 vs. 15.1 months, p = 0.049). There were no significant differences in the mTTF between subgroups based on the starting dose of 40 mg and < 40 mg (16.7 vs. 16.9 months, p > 0.05). The most common treatment-related adverse events (any grade/grade ≥ 3) were diarrhea (55.4%/3.5%), rash (51.9%/3.2%), paronychia (35.3%/5.0%), and stomatitis (22.2%/1.2%). CONCLUSIONS: Afatinib demonstrated clinical effectiveness and good tolerability in Vietnamese EGFR-mutant NSCLC patients. In our real-world setting, administering a starting dose below 40 mg might result in a reduction in ORR; however, it might not have a significant impact on TTF.

Compact UHF Circularly Polarized Multi-Band Quadrifilar Antenna for CubeSat.

This article presents a multi-band right-hand circularly polarized antenna designed for the Cube Satellite (CubeSat). Based on a quadrifilar structure, the antenna provides circular polarization radiation suitable for satellite communication. Moreover, the antenna is designed and fabricated using two 1.6 mm thickness FR4-Epoxy boards connected by metal pins. In order to improve the robustness, a ceramic spacer is placed in the centerboard, and four screws are added at the corners to fix the antenna to the CubeSat structure. These additional parts reduce antenna damage caused by vibrations in the launch vehicle lift-off stage. The proposal has a dimension of 77 × 77 × 10 mm3 and covers the LoRa frequency bands at 868 MHz, 915 MHz, and 923 MHz. According to the measurements in the anechoic chamber, antenna gains with the values of 2.3 dBic and 1.1 dBic are obtained for the 870 MHz and 920 MHz, respectively. Finally, the antenna is integrated into a 3U CubeSat that was launched by a Soyuz launch vehicle in September 2020. The terrestrial-to-space communication link was measured, and the antenna performance was confirmed in a real-life scenario.

The Novel Combination of Nano Vector Network Analyzer and Machine Learning for Fruit Identification and Ripeness Grading.

Fruit classification is required in many smart-farming and industrial applications. In the supermarket, a fruit classification system may be used to help cashiers and customer to identify the fruit species, origin, ripeness, and prices. Some methods, such as image processing and NIRS (near-infrared spectroscopy) are already used to classify fruit. In this paper, we propose a fast and cost-effective method based on a low-cost Vector Network Analyzer (VNA) device augmented by K-nearest neighbor (KNN) and Neural Network model. S-parameters features are selected, which take into account the information on signal amplitude or phase in the frequency domain, including reflection coefficient S11 and transmission coefficient S21. This approach was experimentally tested for two separate datasets of five types of fruits, including Apple, Avocado, Dragon Fruit, Guava, and Mango, for fruit recognition as well as their level of ripeness. The classification accuracy of the Neural Network model was higher than KNN with 98.75% and 99.75% on the first dataset, whereas the KNN was seen to be more effective in classifying ripeness with 98.4% as compared to 96.6% for neural network.

Case report of mild TCIRG1-associated autosomal recessive osteopetrosis in Vietnam.

Autosomal recessive osteopetrosis (ARO) is a group of disease characterized by osteoclast dysfunction inhibiting bone resorption and bone turnover, with TCIRG1-associated ARO being more common leading to autosomal recessive infantile malignant osteopetrosis (OPTB1, MIM entry number # 259700). While most patients with TCIRG1-associated osteopetrosis present a malignant clinical course and shortened lifespan, a few cases of non-malignant TCIRG1-associated osteopetrosis have been reported. 24-year-old female patient came to us with limp gait, hip pain in both sides, and severe stiffness. She had suffered many fractures, bilateral hip osteoarthritis, right leg was 2 cm shorter compared with left leg. Whole Exome Sequencing was conducted, the result and subsequent Sanger's sequencing shown the patient had a compound heterozygous genotype at TCIRG1 (c.1194dup, p.Gly399ArgTer and c.334G>A, p.Gly112Arg), these two variants found were not previously reported. Sanger's sequencing revealed two other siblings whom suffer the same disorder had similar genotype to the proband; the parents were found to be heterozygous. This is the first case of TCIRG1-associated osteopetrosis reported in Vietnam and one of the few cases of nonmalignant TCIRG1-associated osteopetrosis, in which detailed clinical and genetic work-up were performed.

Hyperspectral phasor analysis enables multiplexed 5D in vivo imaging.

Time-lapse imaging of multiple labels is challenging for biological imaging as noise, photobleaching and phototoxicity compromise signal quality, while throughput can be limited by processing time. Here, we report software called Hyper-Spectral Phasors (HySP) for denoising and unmixing multiple spectrally overlapping fluorophores in a low signal-to-noise regime with fast analysis. We show that HySP enables unmixing of seven signals in time-lapse imaging of living zebrafish embryos.

An E3-ligase-based method for ablating inhibitory synapses.

Although neuronal activity can be modulated using a variety of techniques, there are currently few methods for controlling neuronal connectivity. We introduce a tool (GFE3) that mediates the fast, specific and reversible elimination of inhibitory synaptic inputs onto genetically determined neurons. GFE3 is a fusion between an E3 ligase, which mediates the ubiquitination and rapid degradation of proteins, and a recombinant, antibody-like protein (FingR) that binds to gephyrin. Expression of GFE3 leads to a strong and specific reduction of gephyrin in culture or in vivo and to a substantial decrease in phasic inhibition onto cells that express GFE3. By temporarily expressing GFE3 we showed that inhibitory synapses regrow following ablation. Thus, we have created a simple, reversible method for modulating inhibitory synaptic input onto genetically determined cells.

Epidemiological Characteristics of Advanced Hepatocellular Carcinoma in the Northern Region of Vietnam.

Epidemiological characteristics of hepatocellular carcinoma (HCC) in Southern Vietnam has been well reported as in Globocan 2018 while data from the North has still not been fully presented. Therefore, we conducted this retrospective descriptive study on 198 advanced HCC patients treated at 3 major hospitals in Northern Vietnam to describe demographic features, HCC risk factors, and correlation among them in patients with advanced HCC. This information will lead to prevention efforts and provide information for allocating funds for treatment. The median age at diagnosis was 57 years (range: 19-86) and the male/female ratio was 8.9/1. The proportions of hepatitis B virus (HBV) and hepatitis C virus (HCV) infection were 81.3% and 5.6%, respectively. Hepatitis C virus infection rate was significantly higher in patients <50 years old (12.5% vs 3.3%, P = .016). There was no significant difference in age or viral hepatitis infection status by gender. Only 7.6% of patients diagnosed with advanced HCC were asymptomatic. In conclusion, with the high rate of HBV infection among patients with advanced HCC, it is necessary for increasing prevention efforts in HBV screening. Furthermore, HCV infection should be noticed in patients with advanced HCC younger than 50 years old.

Biotagging, an in vivo biotinylation approach for cell-type specific subcellular profiling in zebrafish.

Interrogation of gene regulatory circuits in complex organisms requires precise and robust methods to label cell-types for profiling of target proteins in a tissue-specific fashion as well as data analysis to understand interconnections within the circuits. There are several strategies for obtaining cell-type and subcellular specific genome-wide data. We have developed a methodology, termed "biotagging" that uses tissue-specific, genetically encoded components to biotinylate target proteins, enabling in depth genome-wide profiling in zebrafish. We have refined protocols to use the biotagging approach that led to enhanced isolation of coding and non-coding RNAs from ribosomes and nuclei of genetically defined cell-types. The ability to study both the actively translated and transcribed transcriptome in the same cell population, coupled to genomic accessibility assays has enabled the study of cell-type specific gene regulatory circuits in zebrafish due to the high signal-to-noise achieved via its stringent purification protocol. Here, we provide detailed methods to isolate, profile and analyze cell-type specific polyribosome and nuclear transcriptome in zebrafish.

A versatile gene trap to visualize and interrogate the function of the vertebrate proteome.

We report a multifunctional gene-trapping approach, which generates full-length Citrine fusions with endogenous proteins and conditional mutants from a single integration event of the FlipTrap vector. We identified 170 FlipTrap zebrafish lines with diverse tissue-specific expression patterns and distinct subcellular localizations of fusion proteins generated by the integration of an internal citrine exon. Cre-mediated conditional mutagenesis is enabled by heterotypic lox sites that delete Citrine and "flip" in its place mCherry with a polyadenylation signal, resulting in a truncated fusion protein. Inducing recombination with Cerulean-Cre results in fusion proteins that often mislocalize, exhibit mutant phenotypes, and dramatically knock down wild-type transcript levels. FRT sites in the vector enable targeted genetic manipulation of the trapped loci in the presence of Flp recombinase. Thus, the FlipTrap captures the functional proteome, enabling the visualization of full-length fluorescent fusion proteins and interrogation of function by conditional mutagenesis and targeted genetic manipulation.

Association between maternal depression and risk of infant diarrhea: a systematic review and meta-analysis.

OBJECTIVE: To systematically review the evidence for an association between maternal depression and the risk of diarrheal illness in infants. STUDY DESIGN: This study is a meta-analysis. METHODS: Nine databases were searched systematically for studies that investigated the risk of diarrheal illness in infants born to mothers with depression. RESULTS: Our search strategy yielded 10 studies of the association between maternal depression and the risk of diarrheal illness in infants: eight studies of postnatal depression, two studies of antenatal depression, and one study of perinatal depression. Our meta-analysis showed that infants born to mothers with postnatal depression were more likely to develop diarrheal illness (odds ratio [OR] = 1.902, 95% confidence interval [95% CI] = 1.385-2.612, P < 0.001, n = 18,585). A similar trend was noted for antenatal depression (OR = 2.703, 95% CI = 0.920-7.942, P = 0.071, n = 583). Only one study reported an association between perinatal depression and risk of diarrhea in infants (OR = 1.848, log OR = 0.614, standard error = 0.093, n = 107,587). CONCLUSIONS: Only a few studies so far have identified an association between maternal depression and risk of diarrhea in infants. The evidence available to date suggests that infants born to mothers with depression are more likely to develop diarrhea than infants whose mothers do not have depression.

Control of ZnO Nanorod Defects to Enhance Carrier Transportation in p-Cu2O/i-ZnO Nanorods/n-IGZO Heterojunction.

The p-Cu2O/i-ZnO nanorods/n-IGZO heterojunctions were fabricated by electrochemical and sputtering method. ZnO nanorods were grown on conductive indium gallium zinc oxide (IGZO) thin film and then p-Cu2O layer was deposited on ZnO nanorods to form the heterojunction. ZnO nanorods play an important role in carrier transport mechanisms and performance of the junction. The changing of defects in ZnO nanorods by annealing samples in air and vacuum have studied. The XRD, photoluminescence (PL) spectroscopy, and FTIR were used to study about structure, and defects in ZnO nanorods. The SEM, i­V characteristics methods were also used to define structure, electrical properties of the heterojunctions layers. The results show that the defects in ZnO nanorods affected remarkably on performance of heterojunctions of solar cells.

A comparison of symptoms and quality of life between medial flap and coblation turbinator of inferior turbinate reduction in endoscopic septoturbinoplasty: an analysis of 108 cases.

Introduction: The combination of septoplasty and turbinoplasty is a common surgical and accepted intervention to correct the nasal obstruction. The coblation submucosal reduction turbinator is a new surgical device, and it started to be used recently. On the other hand, the medial flap inferior turbinoplasty is not a conservative technique, but it provides a reliable and robust reduction. Objective: This study aims to compare the symptoms as well as health-related quality of life (HQOL) in 55 patients who underwent septoplasty with concomitent medial flap inferior turbinoplasty (group 1), 53 patients who patients underwent septoplasty with concomitent coblation turbinator (group 2). Patients and methods: The authors performed a prospective, randomized study of 108 patients who consulted the otorhinolaryngology department at the university hospital for surgery of septoturbinoplasty. Results: Preoperatively, the two patient groups had quite similar symptoms and health-related quality of life, and the anterior width of the inferior turbinate showed significant differences between the contralateral and deviated sides but not the posterior part. The significant difference (P<0.05) was noted for postoperatively improved symptom scores on visual analogue scale (VAS), nasal obstruction septoplasty effectiveness (NOSE) and better HQOL [Sino-Nasal Outcome Test-22 (SNOT-22)] in all patient groups. In addition, the NOSE and SNOT-22 scores in group 2 had significantly greater improvement than group 1 (P<0.05). Conclusions: Septoturbinoplasty treatment of septum deviation and inferior turbinate hypertrophy led to less symptoms as well as better HQOL for all two patient groups. Therefore, these techniques were an effective intervention for turbinate reduction, and they are equally efficient in the long term.

Real-world analysis of afatinib as a first-line treatment for patients with advanced stage non-small-cell lung cancer with uncommon EGFR mutations: a multicenter study in Vietnam.

Background: Afatinib is indicated for advanced-stage non-small-cell lung cancer (NSCLC) with Epidermal Growth Factor Receptor (EGFR) and uncommon mutations. However, real-world studies on this topic are limited. This study aimed to evaluate afatinib as first-line therapy for locally advanced and metastatic NSCLC with uncommon EGFR mutations. Patients and methods: A retrospective study included 92 patients with advanced NSCLC with uncommon and compound EGFR mutations, treated with afatinib as first-line therapy. Patients were followed up and evaluated every 3 months or when symptoms of progressive disease arose. The endpoints were objective response rate (ORR), time-to-treatment failure (TTF), and adverse events. Results: The G719X EGFR mutation had the highest occurrence rate (53.3% for both monotherapy and the compound). By contrast, the compound mutation G719X-S768I was observed at a rate of 22.8%. The ORR was 75%, with 15.2% of patients achieving complete response. The overall median TTF was 13.8 months. Patients with the G719X EGFR mutation (single and compound) had a median TTF of 19.3 months, longer than that of patients with other mutations, who had a median TTF of 11.2 months. Patients with compound EGFR mutations (G719X and S768I) demonstrated a median TTF of 23.2 months compared to that of 12.3 months for other mutations. Tolerated doses of 20 or 30 mg achieved a longer median TTF of 17.1 months compared to 11.2 months with 40 mg. Median TTF differed between patients with and without brain metastasis, at 11.2 and 16.9 months, respectively. Rash (55.4%) and diarrhea (53.3%) were the most common adverse events, primarily grades 1 and 2. Other side effects occurred at a low rate. Conclusion: Afatinib is effective for locally advanced metastatic NSCLC with uncommon EGFR mutations. Patients with G719X, compound G719X-S768I mutations, and tolerated doses of 20 or 30 mg had a longer median TTF than those with other mutations.

Enhancer and gene traps for molecular imaging and genetic analysis in zebrafish.

Enhancer and gene trapping methods are highly effective means for the identification and functional analysis of transcriptionally active genes. With recent advances in fluorescent proteins and transposon based integration technologies, a growing family of trapping approaches has been developed in zebrafish, offering powerful tools to both visualize and functionally dissect gene networks during development. Coupled with the intrinsic advantages of the zebrafish model system, creative genetic engineering of trap vectors has enabled high-resolution molecular imaging and genetic manipulations. This review highlights the different enhancer and gene trap approaches that have been developed in zebrafish and offers insights into the strengths, limitations and experimental strategies for their application to enrich our knowledge of gene function and the cellular processes they control.

Dataset of food delivery app users at a public university: Insights into digital transformation.

The food delivery apps (FDAs) have facilitated the connection between food service providers and consumers, enabling online ordering through smartphones and offline delivery in Vietnam. The Covid-19 pandemic has significantly impacted the food and beverage industry, accelerating the process of digital transformation and promoting sustainability through online-to-offline service. The usage of FDAs amongst consumers has exhibited a discernible escalation, primarily attributable to its ability to expedite the delivery of food in a hassle-free and convenient manner. Given the ongoing pandemic and the swift increase in demand for online food ordering services, particularly among the younger demographic, it has become imperative to comprehend the drivers that impel consumers to adopt these applications. This article aims to present a dataset pertaining to the decision-making factors that university students in Danang, Vietnam, take into account when they use FDAs and express positive feedback about them on the internet. The survey was conducted between September 2022 and January 2023 and gathered 346 usable responses. The results provide novel perspectives on the adoption of FDAs by university students, which is an emerging technology in the food and beverage sector. This dataset could be useful to various stakeholders, such as service providers, small and medium enterprises (SMEs), and vendors operating on these platforms, as it can help them acquire valuable insights into their customers' preferences and behavior. In addition, the dataset can serve as a basis for conducting comparative research in different universities or countries.

Hand2 represses non-cardiac cell fates through chromatin remodeling at cis- regulatory elements.

Developmental studies have revealed the importance of the transcription factor Hand2 in cardiac development. Hand2 promotes cardiac progenitor differentiation and epithelial maturation, while repressing other tissue types. The mechanisms underlying the promotion of cardiac fates are far better understood than those underlying the repression of alternative fates. Here, we assess Hand2-dependent changes in gene expression and chromatin remodeling in cardiac progenitors of zebrafish embryos. Cell-type specific transcriptome analysis shows a dual function for Hand2 in activation of cardiac differentiation genes and repression of pronephric pathways. We identify functional cis- regulatory elements whose chromatin accessibility are increased in hand2 mutant cells. These regulatory elements associate with non-cardiac gene expression, and drive reporter gene expression in tissues associated with Hand2-repressed genes. We find that functional Hand2 is sufficient to reduce non-cardiac reporter expression in cardiac lineages. Taken together, our data support a model of Hand2-dependent coordination of transcriptional programs, not only through transcriptional activation of cardiac and epithelial maturation genes, but also through repressive chromatin remodeling at the DNA regulatory elements of non-cardiac genes.

Live imaging of endogenous periodic tryptophan protein 2 gene homologue during zebrafish development.

Yeast Periodic tryptophan protein 2 gene (Pwp2) is involved in ribosome biogenesis and has been implicated in regulation of the cell cycle in yeast. Here, we report a zebrafish protein-trap line that produces fluorescently tagged Periodic tryptophan protein 2 gene homologue (Pwp2h) protein, which can be dynamically tracked in living fish at subcellular resolution. We identified both full-length zebrafish Pwp2h and a short variant. The expression results show that Pwp2h is present in numerous sites in the early developing embryo, but later is restricted to highly proliferative regions, including the forebrain ventricular zone and endoderm-derived organs in the early larval stage. At the subcellular level, Pwp2h protein appears to be localized to the region of the nucleolus consistent with its presumed function in ribosomal RNA synthesis. This Pwp2h protein trap line offers a powerful tool to study the link between ribosome biogenesis and cell cycle progression during vertebrate development.