Tissue expression of porcine transient receptor potential mucolipin protein channels and their differential responses to porcine reproductive and respiratory syndrome virus infection in vitro.

Introduction: Porcine reproductive and respiratory syndrome virus (PRRSV) infection results in a serious disease, posing a huge economic threat to the global swine industry. The transient receptor potential mucolipin proteins (TRPMLs) have been shown to be strongly associated with virus infection and other physiological processes in humans, but their tissue distribution and responses to PRRSV in pigs remain unknown. Material and Methods: Quantitative reverse-transcription PCR analysis was undertaken to determine the optimal primer for TRPML expression detection and for quantifying that expression individually in different pig tissue samples. Meat Animal Research Center 145 (MARC-145) monkey kidney cells and the TRPML-specific activator mucolipin synthetic agonist 1 (ML-SA1) were used to reveal the relationship between TRPML and PRRSV-2 infection. Results: The best primers for each TRPML gene used in a fluorescence-based quantitative method were identified and TRPML1 was found to be highly expressed in the kidneys and liver of pigs, while TRPML2 and TRPML3 were observed to be primarily expressed in the kidney and spleen tissues. The expression of TRPML2 was upregulated with the rise in PRRSV-2 infection titre but not the expression of TRPML1 or TRPML3, and ML-SA1 inhibited PRRSV-2 in a dose-dependent manner. Conclusion: Our research revealed the gene expression of TRPMLs in pigs and identified that TRPML channels may act as key host factors against PRRSV infection, which could lead to new targets for the prevention and treatment of pig infectious diseases.

Comprehensive single-cell analysis reveals heterogeneity of fibroblast subpopulations in ovarian cancer tissue microenvironment.

Background: Ovarian cancer, as a highly malignant tumor, features the critical involvement of tumor-associated fibroblasts in the ovarian cancer tissue microenvironment. However, due to the apparent heterogeneity within fibroblast subpopulations, the specific functions of these subpopulations in the ovarian cancer tissue microenvironment remain insufficiently elucidated. Methods: In this study, we integrated single-cell sequencing data from 32 ovarian cancer samples derived from four distinct cohorts and 3226 bulk RNA-seq data from GEO and TCGA-OV cohorts. Utilizing computational frameworks such as Seurat, Monocle 2, Cellchat, and others, we analyzed the characteristics of the ovarian cancer tissue microenvironment, focusing particularly on fibroblast subpopulations and their differentiation trajectories. Employing the CIBERSORTX computational framework, we assessed various cellular components within the ovarian cancer tissue microenvironment and evaluated their associations with ovarian cancer prognosis. Additionally, we conducted Mendelian randomization analysis based on cis-eQTL to investigate causal relationships between gene expression and ovarian cancer. Results: Through integrative analysis, we identified 13 major cell types present in ovarian cancer tissues, including CD8+ T cells, malignant cells, and fibroblasts. Analysis of the tumor microenvironment (TME) cell proportions revealed a significant increase in the proportion of CD8+ T cells and CD4+ T cells in tumor tissues compared to normal tissues, while fibroblasts predominated in normal tissues. Further subgroup analysis of fibroblasts identified seven subgroups, with the MMP11+Fib subgroup showing the highest activity in the TGFß signaling pathway. Single-cell analysis suggested that oxidative phosphorylation could be a key pathway driving fibroblast differentiation, and the ATRNL1+KCN + Fib subgroup exhibited chromosomal copy number variations. Prognostic analysis using a large sample size indicated that high infiltration of MMP11+ fibroblasts was associated with poor prognosis in ovarian cancer. SMR analysis identified 132 fibroblast differentiation-related genes, which were linked to pathways such as platinum drug resistance. Conclusions: In the context of ovarian cancer, fibroblasts expressing MMP11 emerge as the primary drivers of the TGF-beta signaling pathway. Their presence correlates with an increased risk of adverse ovarian prognoses. Additionally, the genetic regulation governing the differentiation of fibroblasts associated with ovarian cancer correlates with the emergence of drug resistance.

Exploring the relationship between digital transformation and green innovation: The mediating role of financing modes.

Green innovation is important and beneficial for the green development of enterprises. The development of digital technology could improve the asymmetry of information and affect the enterprise's financing modes. Currently, a definitive conclusion about the relationship between the digital transformation and green technology innovation has not reached. Additionally, studies have not yet fully explored the impact pathways of corporate digital transformation on green innovation through corporate financing modes. To fill this void, this study collected data of Chinese A-share listed companies from 2010 to 2020. The empirical analysis results showed that digital transformation significantly enhanced enterprises' green innovation abilities. Furthermore, mediating models were utilized to explore how enterprise financing modes mediate the impact of digital transformation on the enterprise green innovation. Results indicated that through internal financing and bank loans as "bridges", digital transformation promoted enterprise green innovation. However, digital transformation did not have a significant impact on enterprise green innovation through venture capital. Moreover, heterogeneity analyses revealed significant differences among different groups of enterprises. This study expands the research field related to digital transformation and enterprise green innovation by providing empirical evidence to uncover the impact of digital transformation on enterprise green innovation. Findings of this study contribute not only to enterprise managerial decision-makers to better leverage digital transformation through corporate financing to accelerate green innovation, but also to government policymakers to design policies to stimulate enterprise green innovation.

Differences in Sleep EEG Coherence and Spindle Metrics in Toddlers With and Without Language Delay: A Prospective Observational Study.

Background: Sleep plays a crucial role in early language development, and sleep disturbances are common in children with neurodevelopmental disorders. Examining sleep microarchitecture in toddlers with and without language delays can offer key insights into neurophysiological abnormalities associated with atypical neurodevelopmental trajectories and potentially aid in early detection and intervention. Methods: Here, we investigated electroencephalogram (EEG) coherence and sleep spindles in 16 toddlers with language delay (LD) compared with a group of 39 typically developing (TD) toddlers. The sample was majority male (n = 34, 62%). Participants were aged 12-to-22 months at baseline, and 34 (LD, n=11; TD, n=23) participants were evaluated again at 36 months of age. Results: LD toddlers demonstrated increased EEG coherence compared to TD toddlers, with differences most prominent during slow-wave sleep. Within the LD group, lower expressive language skills were associated with higher coherence in REM sleep. Within the TD group, lower expressive language skills were associated with higher coherence in slow-wave sleep. Sleep spindle density, duration, and frequency changed between baseline and follow-up for both groups, with the LD group demonstrating a smaller magnitude of change than the TD group. The direction of change was frequency-dependent for both groups. Conclusions: These findings indicate that atypical sleep EEG connectivity and sleep spindle development can be detected in toddlers between 12 and 36 months and offers insights into neurophysiological mechanisms underlying the etiology of neurodevelopmental disorders. Trial registration: https://clinicaltrials.gov/study/NCT01339767; Registration date: 4/20/2011.

New Alpiniamide-Type Polyketide with Antibiofilm Activities from the Marine-Derived Streptomyces sp. ZS-A65.

Two new alpiniamide-type polyketides, alpiniamides H-I (1-2), in addition to four recognized compounds, were discovered in Streptomyces sp. ZSA65 derived from the marine sediments. The planar structure and absolute configuration of alpiniamides H-I were elucidated using a combination of 1D, 2D NMR, HRESIMS data analysis, Mosher's method and ECD calculations. The antibiofilm and antibacterial activities against P. aeruginosa were evaluated using the microdilution method. Notably, Compound 2 exhibited strong antibiofilm property.

Combined use of Anlotinib with chemotherapy in patients with advanced ovarian cancer: a real-world cohort study and meta-analysis.

Background: Anlotinib is a novel oral small-molecule receptor tyrosine kinase inhibitor. However, the efficacy and safety of its combined use with chemotherapy remain unclear in patients with advanced ovarian cancer. Objectives: To assess the efficacy and safety of the combined use of Anlotinib with chemotherapy in patients with advanced ovarian cancer. Design: A multi-center retrospective real-world analysis and a meta-analysis. Data sources and methods: We enrolled patients with advanced ovarian cancer who received a combination therapy of Anlotinib and chemotherapy from 15 medical centers. We also searched electronic databases for studies assessing the efficacy and safety of the combined use of Anlotinib with chemotherapy in patients with ovarian cancer. The outcomes of interest included objective response rate (ORR), disease control rate (DCR), and median progression-free survival (mPFS). Results: A total of 71 patients, who were predominantly recurrent cases, were included in the real-world study. The ORR and DCR of the included patients were 40.8% and 76.1%, respectively; and their mPFS was 4.6 months. The log-rank test showed that previous antiangiogenic therapy was related to a longer mPFS (p < 0.05). Five studies in total were eligible for meta-analysis. The random-effects meta-analysis model showed that the ORR, DCR, and mPFS were 33.8% [95% confidence interval (CI) 22.7-44.8% from four studies], 90.6% (95% CI 73.6-99.9% from five studies), and 6.6 months (95% CI 4.9-8.4 months from five studies). The most common adverse events were hand-foot syndrome and hypertension. Conclusion: The combined use of Anlotinib with chemotherapy showed potential in treating patients with advanced ovarian cancer, with a tolerable safety profile.

Exosomes derived from ovarian cancer cells regulate proliferation and migration of cancer-associated fibroblasts.

Cancer-associated fibroblast (CAF) is an essential risk factor for ovarian cancer. Exosomes can mediate cellular communication in the tumour microenvironment, but the interaction of tumour cell exosomes with CAF is less studied in Ovarian cancer. This study identified H19/miR-29c-3p/LOXL2-COL1A1 as a ceRNA regulatory network involved in regulating tumour matrix-associated signaling pathways associated with CAF. Cellular assays demonstrated that exosomes from ovarian cancer cell line SKOV3 significantly promoted the proliferation and migration of CAF. The results of mixed transplantation tumour experiments in nude mice showed that exosomes of SKOV3 significantly promoted tumour growth. Ovarian cancer tumour-derived exosomes can regulate CAF proliferation and migration through H19/miR-29c-3p/LOXL2-COL1A1. This study reveals the regulatory role of tumour exosomes on CAF, which may provide a theoretical basis for the development of therapeutic regimens targeting fibroblasts in ovarian cancer.

Efficacy and Safety of Anlotinib in Overall and Disease-Specific Advanced Gynecological Cancer: A Real-World Study.

Purpose: Anlotinib is a novel oral small-molecule multi-target tyrosine kinase inhibitor that has been approved for treating non-small cell lung cancer. However, its efficacy and safety among patients with advanced gynecological cancer have not been comprehensively evaluated. We conducted this study to address this issue in the real-world setting. Patients and Methods: Data from patients treated with Anlotinib for persistent, recurrent or metastatic gynecological cancer were collected from 17 centers from August 2018. The database lock-time was on March 2022. Anlotinib was administered orally on days 1-14 every 3 weeks until disease progression, severe toxicity occurred, or death. In this study, disease-specific advanced gynecological cancer was mainly referred to cervical, endometrial, and ovarian cancer. The outcomes included objective response rate (ORR), disease control rate (DCR), and progression-free survival (PFS). Results: A total of 249 patients were analyzed, with a median follow-up of 14.5 months. The overall ORR and DCR were 28.1% [95% confidence interval (CI) 22.6% to 34.1%] and 80.7% (95% CI 75.3% to 85.4%), respectively. Specifically, the ORR varied from 19.7% to 34.4% and the DCR differed from 81.7% to 90.0% in disease-specific advanced gynecological cancer. The median PFS was 6.1 months and ranged from 5.6 to 10.0 months in the overall and disease-specific advanced gynecological cancer, respectively. Larger cumulative dosage of Anlotinib (>700 mg) was in general associated with longer PFS in the overall and disease-specific advanced gynecological cancer. The most common adverse event related to Anlotinib treatment was pain/arthralgia (18.3%). Conclusion: In conclusion, Anlotinib holds promise in treating patients with advanced gynecological cancer including its disease-specific types, with reasonable efficacy and tolerable safety.

Detection and Separation of DNA and Silver Nanoparticles Using a Solid-State Nanopore.

Nanopore sensors, a new generation of single-molecule sensors, are increasingly used to detect and analyze various analytes and have great potential for rapid gene sequencing. However, there are still some problems in the preparation of small diameter nanopores, such as imprecise pore size and porous defects, while the detection accuracy of large-diameter nanopores is relatively low. Therefore, how to achieve more precise detection of large diameter nanopore sensors is an urgent problem to be studied. Here, SiN nanopore sensors were used to detect DNA molecules and silver nanoparticles (NPs) separately and in combination. The experimental results show that large-size solid-state nanopore sensors can identify and discriminate between DNA molecules, NPs, and NP-bound DNA molecules clearly according to resistive pulses. In addition, the detection mechanism of using NPs to assist in identifying target DNA molecules in this study is different from previous reports. We find that silver NPs can simultaneously bind to multiple probes and target DNA molecules and generate a larger blocking current than free DNA molecules when passing through the nanopore. In conclusion, our research indicates that large-sized nanopores can distinguish the translocation events, thereby identifying the presence of the target DNA molecules in the sample. This nanopore-sensing platform can produce rapid and accurate nucleic acid detection. Its application in medical diagnosis, gene therapy, virus identification, and many other fields is highly significant.

Gene expression of TRPMLs and its regulation by pathogen stimulation.

The transient receptor potential mucolipin (TRPML) subfamily in mammalian has three members, namely TRPML1, TRPML2, and TRPML3, who play key roles in regulating intracellular Ca2+ homeostasis, endosomal pH, membrane trafficking and autophagy. Previous studies had shown that three TRPMLs are closely related to the occurrence of pathogen invasion and immune regulation in some immune tissues or cells, but the relationship between TRPMLs expression and pathogen invasion in lung tissue or cell remains elusive. Here, we investigated the expression distribution of three TRPML channels in mouse different tissues by qRT-PCR, and then found that all three TRPMLs were highly expressed in the mouse lung tissue, as well as mouse spleen and kidney tissues. The expression of TRPML1 or TRPML3 in all three mouse tissues had a significant down-regulation after the treatment of Salmonella or LPS, but TRPML2 expression showed a remarkable increase. Consistently, TRPML1 or TRPML3 but not TRPML2 in A549 cells also displayed a decreased expression induced by LPS stimulation, which shared a similar regulation pattern in the mouse lung tissue. Furthermore, the treatment of the TRPML1 or TRPML3 specific activator induced a dose-dependent up-regulation of inflammatory factors IL-1ß, IL-6 and TNFα, suggesting that TRPML1 and TRPML3 are likely to play an important role in immune and inflammatory regulation. Together, our study identified the gene expression of TRPMLs induced by pathogen stimulation in vivo and in vitro, which may provide novel targets for innate immunity or pathogen regulation.

The Effects of Nutrition and Health Claim Information on Consumers' Sensory Preferences and Willingness to Pay.

As marketing tools, nutrition claims (NCs) and health claims (HCs) can be used to convey the nutritional properties and health benefits of food to consumers, but their respective effects on consumers' perceptions of healthier meat products are inconsistent in the literature. Using a physical prototype of omega-3-enriched sausages as a research interest, this paper explores how HCs and NCs differently influence consumers' sensory preferences and willingness to pay (WTP). Sensory tests were carried out among 330 participants, followed by a choice-based conjoint (CBC) experiment to measure consumers' WTP. Results indicate that, in comparison with the uninformed condition, labeling an omega-3 nutrition claim increased consumers' sensory liking for omega-3-enriched sausages in the attributes of appearance and texture. Moreover, consumers were willing to pay more for healthier sausages, but labeling HCs did not significantly improve participants' WTP for omega-3-enriched sausages more than NCs. Hence, HCs did not significantly outperform NCs, when it comes to positively influencing consumers' sensory liking and paying intentions for omega-3-enriched sausages. The findings of this study have implications for the meat industry in developing healthier sausage formulations with greater likelihood of success in the market.

Laparo-Endoscopic Single Site Combined With Hysteroscopy to Diagnose and Treat Robert's Uterine Malformation: A Case Report.

Asymmetric septate uterus, commonly known as Robert's uterus, is an exceedingly rare uterine malformation described for the first time in 1970 by Robert H. Currently, surgery is the therapy of choice for Robert's uterus, with surgical choices ranging from laparotomy to minimally invasive surgery. In this paper, we reported that a 14-year-old girl with primary dysmenorrhea that gradually worsened three months after menarche had surgery after many imaging evaluations, and that the intraoperative diagnosis was Robert's uterus. The diagnostic and therapeutic laparo-endoscopic single site(LESS) combined with hysteroscopy surgery for Robert's uterine abnormality was shown via a step-by-step presentation of the method accompanied by narrated video footage. During the ten-month postoperative follow-up period, the patient had monthly recurrences with normal menstrual volume and no dysmenorrhea, demonstrating that as a minimally invasive treatment, LESS combined with hysteroscopy surgery is a successful methodfor diagnosing and treating this specific malformation.

A network analysis of internet gaming disorder symptoms.

We examined the symptoms network of Internet Gaming Disorder (IGD). We recruited 793 heavy game players online and 184 of them were diagnosed with IGD. We measured IGD via the 27-item IGD Scale and analyzed symptoms to examine how they interacted with each other to drive the addictive behaviors. Furthermore, we added motives and psychological well-being variables into the network. We found that the most central symptoms were conflict, withdrawal, and tolerance. Overall, two distinct groups of symptoms were discovered: one group for addictive symptoms and the other for psychological consequences.

Network analysis of physical and psychiatric symptoms of hospital discharged patients infected with COVID-19.

In the current study, we aimed to investigate the network structure of COVID-19 symptoms and its related psychiatric symptoms, using a network approach. Specifically, we examined how COVID-19 symptoms relate to psychiatric symptoms and highlighted potential pathways between COVID-19 severity and psychiatric symptoms. With a sample of six hundred seventy-five recovered COVID-19 patients recruited 1 month after hospital discharge, we respectively integrated COVID-19 symptoms with PTSD, depression, and anxiety symptoms and analyzed the three network structures. In all three networks, COVID-19 severity and ICU admission are not linked directly to COVID-19 symptoms after hospitalization, while COVID-19 severity (but not ICU admission) is linked directly to one or more psychiatric symptoms. Specific pathways between COVID-19 symptoms and psychiatric symptoms were discussed. Finally, we used directed acyclic graph estimation to show potential causal effects between COVID-19 related variables and demographic characteristics.

Dual-fluorescent bacterial two-hybrid system for quantitative Protein-Protein interaction measurement via flow cytometry.

Characterization of protein-protein interactions (PPIs) is essential for understanding cellular signal transduction pathways. However, quantitative measurement of the binding strength remains challenging. Building upon the classical bacterial adenylate cyclase two-hybrid (BACTH) system, we previously demonstrated that the relative reporter protein expression (RRPE), defined as the level of reporter expression normalized to that of the interacting protein, is an intrinsic characteristic associated with the binding strength between the two interacting proteins. In this study, we inserted fluorescent protein tdTomato in the chromosome as the reporter protein by CRISPR/Cas9 technology and employed a 12-amino acid tetracysteine (TC) to tag one of the interacting proteins, which can be further labeled by a membrane-permeable biarsenical dye. The combined use of tdTomato and TC-tag offers rapid and high-throughput analysis of the expression levels of both the reporter protein and one of the interacting proteins at the single-cell level by multicolor flow cytometry, which simplifies the quantitative measurement of PPI. The use of the as-developed RRPE-tdTomato-TC-BACTH approach was demonstrated in three demanding applications. First, binding affinities could be correctly ranked for discriminating interaction strengths with a tenfold difference or of the same order of magnitude. We demonstrate that the method is sensitive enough to discriminate affinities with a small difference of 1.4-fold. Moreover, residues involved in PPI can be easily mapped and ranked. Lastly, protein interaction inhibitors can be rapidly screened.

Multiplexed detection of bacterial pathogens based on a cocktail of dual-modified phages.

Rapid, quantitative, and sensitive assays for the multiplexed detection of bacterial pathogens are urgently needed for public health. Here, we report the generation of dual-modified phage sensors for the simultaneous detection of multiple pathogenic bacteria. The M13KE phage was dual modified to display the targeting peptide on the minor coat protein pIII (∼5 copies) and the streptavidin-binding (StrB) peptide on the major coat protein pVIII (∼2700 copies). The targeting peptide specifically recognizes the target bacteria, and the StrB peptide acts as the efficient signal amplification and transduction unit upon binding with fluorescently tagged streptavidin. The bright fluorescence emitted from individual target bacteria can be clearly distinguished from the background via both the flow cytometry and fluorescence microscopy. Three different dual-modified phages targeting E. coli O157:H7, Salmonella Typhimurium, and Pseudomonas aeruginosa were constructed, and high specificity was verified via a large excess of other non-target bacteria. Using a 40 mL sample volume, the target bacteria detection limit was approximately 102 cells/mL via flow cytometry measurement in the presence of other non-target bacteria. By combining these three dual-modified phages into a cocktail, simultaneous detection and quantification of three target bacterial pathogens was demonstrated with good linearity. The strategy of constructing dual-modified phage represents a promising tool in the detection of bacterial pathogens.

Label-Free Detection of Bacteria in Fruit Juice by Nano-Flow Cytometry.

Rapid quantification of microbial contamination in fruit juice is highly desired for food safety control. Yet, the complex sample matrix and the diversity of bacterial contaminants present a great challenge. Employing a laboratory-built nano-flow cytometer (nFCM), here we report the development of a label-free approach for the detection of bacteria population in fruit juice. The weak autofluorescence of bacterial cells was used as a hallmark for the identification of bacteria. The sample pretreatment protocol was optimized to reduce fluorescence background, lyse residual plant cells and debris, and attain a good recovery of bacteria from juice samples. It was demonstrated that the nFCM was able to enumerate individual bacteria of very weak autofluorescence, and a clear differentiation from residual juice particulates was achieved. For bacteria spiked in the orange juice, the recovery rate was around 95% and a linear correlation between nFCM analysis and plate counting was acquired in the range of 3 × 104 to 3 × 108 cfu/mL. The assay, including sample pretreatment and instrument analysis, can be accomplished within 1 h, which is far more efficient than plate counting. Using a 40 mL sample volume, the detection limit in apple juice was ∼102 cells/mL. The as-developed method was successfully applied to bacterial measurement of freshly made orange juice and apple juice purchased from grocery stores. We believe it could also have potential practical application in microbial control analysis of other juices and water.

The efficacy of virtual reality exposure therapy for PTSD symptoms: A systematic review and meta-analysis.

BACKGROUND: Virtual reality exposure therapy (VRET) for PTSD is an emerging treatment of remarkable promise, but its efficacy and safety are still unclear. Our aim was to investigate the efficacy of VRET for individuals with PTSD, and to identify the potential moderating variables associated with interventions. METHODS: Literature search was conducted via PubMed, Embase, Web of Science, Cochrane Library, PsycInfo, Science Direct, and EBSCO. We identified 18 studies on PTSD including 13 randomized controlled trials (RCTs; 654 participants) and 5 single-group trials (60 participants). RESULTS: The main effects analysis showed a moderate effect size (g = 0.327, 95% CI: 0.105-0.550, p<0.01) for VRET compared to control conditions on PTSD symptoms. Subgroup analysis revealed that the effects of VRET were larger when compared to inactive groups (g = 0.567) than active control groups (g = 0.017). This finding was in agreement with depressive symptoms. A dose-response relationship existed with more VRET sessions showing larger effects. There was a long-range effect of VRET on PTSD symptoms indicating a sustained decrease in PTSD symptoms at 3-month follow-up (g = 0.697) and 6-month follow-up (g = 0.848). The single-group trials analysis revealed that the VRET intervention had a significant effect on PTSD. LIMITATIONS: Many of the combat-related PTSD subjects resulted in uncertainty regarding meta-analytical estimates and subsequent conclusions. CONCLUSIONS: These findings demonstrated that VRET could produce significant PTSD symptoms reduction and supported its application in treating PTSD.

The Conceptual Modeling of Interoperability Framework of Heart Sound Monitor in the Context of an Interoperable End-to-End Architecture.

Background: Heart sound monitor (HSM), a device suitable for home-use, can be used to acquire heart sounds. It enables the telemonitoring of cardiac function, which has been largely evolved and widely used in recent years. Nevertheless, the designers paid little attention to the consistency of information model and data interaction of HSM, thus the data could not be shared and aggregated among healthcare systems. Consequently, the device's development and its application in person-centered telehealth are hindered. Objective: To solve this problem and to build interoperability for HSM, this article proposes a HSM interoperability framework that is constructed by using standardized modeling methods. Methods: The authors collected the common device-output information of HSM involved in telemonitoring, leveraged the standardized interoperability framework defined in ISO/IEEE 11073 Personal Health Device (11073-PHD) standards to model the static data structure and dynamic interaction behaviors of HSM. Results: Via a meta-analysis, the HSM device-output information includes collected data (heart sound measurement), and derived data (e.g., device status). Based on such information, an 11073-PHD-compliant domain information model has been successfully created. This enables the interoperability between HSM and aggregation device, allowing inter-device plug-and-play using the service model and communication model. A prototype of this design has been implemented and validated via Continua Enabling Software Library. Conclusions: The ISO/IEEE 11073-PHD standard framework has the potential to accommodate the HSM, which implicate HSM can be integrated into the interoperable ecosystem to achieve holistic health solution. Findings in this article may be taken as a reference for standard developing organizations to establish a standardized interoperability framework for HSM.

Rapid quantification of live/dead lactic acid bacteria in probiotic products using high-sensitivity flow cytometry.

A laboratory-built high-sensitivity flow cytometer (HSFCM) was employed for the rapid and accurate detection of lactic acid bacteria (LAB) and their viability in probiotic products. LAB were stained with both the cell membrane-permeable SYTO 9 green-fluorescent nucleic acid stain and the red-fluorescent nucleic acid stain, propidium iodide, which penetrates only bacteria with compromised membranes. The side scatter and dual-color fluorescence signals of single bacteria were detected simultaneously by the HSFCM. Ultra-high temperature processing milk and skim milk spiked with Lactobacillus casei were used as the model systems for the optimization of sample pretreatment and staining. The viable LAB counts measured by the HSFCM were in good agreement with those of the plate count method, and the measured ratios between the live and dead LAB matched well with the theoretical ratios. The established method was successfully applied to the rapid quantification of live/dead LAB in yogurts and fermented milk beverages of different brands. Moreover, the concentration and viability status of LAB in ambient yogurt, a relatively new yet popular milk product in China, are also reported.