Therefore, the data suggest that LINC01977 interacts with RBM39 and promotes the progression of HCC by inhibiting Notch2 ubiquitination and degradation, indicating that LINC01977 may be a potential biomarker and therapeutic target for HCC patients.Cancer-testis genes are involved in the occurrence and development of cancer, but the role of cancer-testis-associated lncRNAs (CT-lncRNAs) in hepatocellular carcinoma (HCC) remains to be explored. Here, we discovered a novel CT-lncRNA, LINC01977, based on the Genotype-Tissue Expression (GTEx) and The Cancer Genome Atlas (TCGA) databases. LINC01977 was exclusively expressed in testes and highly expressed in HCC. High LINC01977 levels correlated with poorer overall survival (OS) in individuals with HCC. Functional assays showed that LINC01977 promoted HCC growth and metastasis in vitro and in vivo. Mechanistically, LINC01977 directly bound to RBM39 to promote the further entry of Notch2 into the nucleus, thereby preventing the ubiquitination and degradation of Notch2. Furthermore, the RNA binding protein IGF2BP2, one of the m Hepatocellular carcinoma (HCC), the main type of primary liver cancer, is a common malignant tumor of the digestive system . AccordiCancer-testis (CT) genes, as the name suggests, are usually expressed only in testis tissue, with little or very low expression in other tissues . HoweverAn increasing number of investigations have revealed that lncRNAs exert an irreplaceable role in different cancers –18. The Here, we discovered another conserved CT-lncRNA, LINC01977, the function of which has not been reported in liver cancer. We found that CT-lncRNA LINC01977 played a key role in the IGF2BP2-LINC01977-RBM39-Notch2 axis, leading to the development of HCC.LINC01977 is a cancer-testis gene obtained from the GTEx and TCGA databases. In normal human tissues, LINC01977 is highly expressed in the testis, while its expression is very low or essentially absent in other normal tissues Fig. . Then, wP = 0.026), more tumor number (P = 0.003), vascular invasion (P = 0.004), and advanced grade (P = 0.000)  and LINC01977 overexpression cell lines (SMMC7721 and Hep3B) and verified the efficiencies of knockdown and overexpression using qRT-PCR . We found that nucleotides 959–1030 of LINC01977 and amino acids 301–352 of RBM39 were the most likely binding regions   Fig. . The top05) Fig. . Gene se05) Fig. . Notch2,05) Fig. . The tarES6 Fig. . SurprisES6 Fig. . OverexpES6 Fig. . To elucES6 Fig. . MG132, ES6 Fig. . Taken tHEY1 and HES6 . LINC01977 has this modification site, indicating that IGF2BP2 may have a stabilizing effect on LINC01977 in HCC cells. In view of this, we first verified the knockdown efficiency of IGF2BP2 in MHCC97H cells , demethylases (erasers), and effector proteins (readers) [6A-modified transcripts. They play important roles in various aspects such as RNA stability, translation efficiency, RNA splicing, and RNA export [6A enrichment of LINC01977 is very high in HCC cell lines. Analysis of the MS results and experiments both reveals that LINC01977 binds to IGF2BP2. The stability of LINC01977 is significantly decreased after knockdown of IGF2BP2. Therefore, IGF2BP2 acts as an m6A reader and enhances the stability of LINC01977.It has been reported that RNA has many modifications including mfication –38. The readers) . Among tA export , 40–42. Overall, our study demonstrates LINC01977 as a CT-lncRNA. LINC01977 is exclusively expressed in testes and highly expressed in HCC, which promotes HCC progression. Mechanistic studies reveal that IGF2BP2 enhances the stability of LINC01977, resulting in its high level. Meanwhile, LINC01977 binds to RBM39, promotes the further entry of Notch2 into the nucleus, and prevents the ubiquitination and degradation of Notch2. Therefore, LINC01977 exerts a key role in the occurrence and development of HCC through the IGF2BP2-LINC01977-RBM39-Notch2 axis, which suggests that LINC01977 could be a promising target for patients with HCC.Fresh tumor tissues, adjacent noncancerous tissues and normal tissues were all acquired from the Affiliated Drum Tower Hospital, Medical School of Nanjing University. None of the HCC patients received chemotherapy or radiotherapy before resection, and all participating patients signed the relevant informed consent before surgery. All tissues after surgical resection were immediately put into liquid nitrogen for rapid freezing. Approval for this study was obtained from the Institutional Ethics Committee of the Affiliated Drum Tower Hospital, Medical School of Nanjing University. All researches were in compliance with government policies and the Declaration of Helsinki.2 at 37 °C. All cell lines were routinely monitored for mycoplasma and were negative.The human cell lines  were purchased from the cell bank of the Chinese Science Academy. HepG2 and HEK293T cells were purchased from the American Type Culture Collection (ATCC). Cells were cultured in Dulbecco’s modified Eagle’s medium (Invitrogen Life Technologies) supplemented with 10% fetal bovine serum and incubated in a humidified incubator containing 5% COShort hairpin RNA (shRNA) targeting LINC01977 was purchased from GenePharma. The lentivirus overexpressing LINC01977 was purchased from Vigene. Briefly, lentiviral packaging required cotransfection of target and packaging plasmids (psPAX2 and pMD2.G) into HEK293T cells using transfection reagents. After 48 h, the supernatant containing lentiviral particles was collected and filtered. Cells were infected with lentiviruses and screened for positively transfected cells with puromycin. Finally, the efficiency of transfection was verified by qRT-PCR. All sequences are listed in Supplementary Table Colony formation assays were performed by seeding cells in six-well plates at 400 cells per well and placing them in an incubator for 10 days. Then, the cells were stained with crystal violet (Beyotime) and counted. Cell viability was assessed using the EdU assay (RiboBio). Soft agar colony formation assays required different concentrations of agar solutions followed by HCC cell suspension preparation and incubator placement for 14 days. For the HCC organoid assay, HCC organoids were first established with human HCC fresh tissues, and then the organoids were transfected with LINC01977-overexpressing lentivirus and control lentivirus. After 14 days, images of HCC organoids were obtained by light microscopy.HCC cells were first added to six-well plates and placed in an incubator. Then, the supernatant was collected and filtered through a 0.22-micron filter. Filtered supernatants were used to resuspend HUVECs, and the cells were seeded in 50 μl Matrigel-coated 96-well plates. After 6 h, the tubular structures were observed with a microscope and photographed for analysis.For the Transwell assay, the upper chamber was seeded with HCC cells in serum-free medium, with or without Matrigel (BD Biosciences). The lower chamber was filled with 700 μl of medium containing 10% FBS, and the cells were placed in an incubator for 48 h. Then, the cells were stained with crystal violet (Beyotime) and counted.TRIzol reagent (Invitrogen Life Technologies) was utilized to obtain RNA from tissues and cells. cDNA was derived from one microgram of RNA using HiScript Q RT SuperMix (Vazyme). qRT-PCR was carried out with the SYBR Green PCR Kit (Vazyme) and detected by the Applied Biosystems 7900HT Sequence Detection System (Applied Biosystems). Primers are listed in Supplementary Table Cell or tissue proteins were extracted using RIPA lysis buffer with protease and phosphatase inhibitors. The protein concentration was determined by a BCA Protein Assay Kit (Beyotime). Approximately 30 μg of protein was loaded per lane. The proteins were separated by 10% SDS-PAGE and transferred to a 0.45 μm polyvinylidene fluoride (PVDF) membrane. The primary antibody was incubated with the membrane overnight at 4 °C. The membranes were washed with TBST, incubated with the corresponding secondary antibodies for 2 h at room temperature, and then washed with TBST. Finally, the target protein bands were detected using ECL detection reagent. The antibodies used are listed in Supplementary Table Actinomycin D (MCE) was added to cells to restrain de novo RNA synthesis. At different time points, cells were harvested with TRIzol reagent. Finally, total cellular RNA was acquired and qRT-PCR was carried out for further analysis.Eight-micrometer-thick paraffin-embedded tissue was transferred to a glass slide. The slides were then deparaffinized and hydrated. Tris-EDTA pH 9.0 was utilized for antigen retrieval. Intrinsic peroxidase activity was inactivated and serum blocking was performed at room temperature. Ki67 antibody  and RBM39 antibody  were added to the tissues and incubated for 13 h at 4 °C. Next, the slides were washed and biotinylated secondary antibody was added to the slides for incubation. Finally, the tissues were colored with DAB solution, dehydrated with ethanol and mounted. Light microscopy was used to take photos and analyze the samples.For tissue immunofluorescence, slides were deparaffinized and hydrated. Tris-EDTA pH 9.0 was utilized for antigen retrieval. Intrinsic peroxidase activity was inactivated, and serum blocking was performed at room temperature. CD31, E-cadherin, Vimentin and RBM39 antibodies were added to the tissues and incubated for 13 h at 4 °C. Antibody concentrations were all diluted 1:200. Next, the slides were washed and Alexa Fluor-labeled secondary antibodies (Abcam) were added to the slides for incubation. Finally, the tissues were stained with DAPI solution for nuclei and photographed with a confocal microscope (Leica).For cellular immunofluorescence, cells were first fixed with 4% paraformaldehyde. Then, serum blocking was performed at room temperature. E-cadherin, Vimentin and RBM39 antibodies were added to the cells and incubated for 13 h at 4 °C. Antibody concentrations were all diluted 1:200. Next, the cells were washed and Alexa Fluor-labeled secondary antibodies (Abcam) were added to the cells. Finally, the cells were stained with DAPI solution for nuclei and photographed using a confocal microscope (Leica).Cytoplasmic and nuclear RNAs were separated and purified from MHCC97H and HepG2 cells using the Cytoplasmic and Nuclear RNA Purification Kit (Norgen). The RNAs were then reverse transcribed into cDNAs, and qRT-PCR was utilized to determine the expression levels of LINC01977, GAPDH and U2 in the cytoplasm and nucleus. GAPDH and U2 were used as cytoplasmic and nuclear controls, respectively.MHCC97H and HepG2 cells were subjected to FISH using an RNA FISH kit (RiboBio). The target probe LINC01977 and control probes (18 S and U6) were supplied by RiboBio and labeled with Cy3. Firstly, cells were seeded in confocal dishes. When the cells are completely attached, a series of subsequent operations such as fixation and permeabilization were carried out. The prehybridization solution was used for blocking. The corresponding probes were utilized for overnight incubation to allow the probes to bind to the targets. Finally, the cells were stained with DAPI solution for nuclei and photographed using a confocal microscope (Leica).The full-length LINC01977 plasmid was cloned into the pGEM-3Z vector. First, linear plasmids were cut enzymatically and transcribed into biotinylated RNA in vitro using T7 and Sp6 RNA polymerases. Magnetic beads, biotinylated RNA, and MHCC97H cell lysates were coincubated at 4 °C using a Pierce Magnetic RNA-Protein Pull-Down kit (Thermo Fisher Scientific). Finally, proteins were eluted and subjected to mass spectrometry and western blotting analyses.6A (Abcam) and IGF2BP2 (Proteintech). Purified RNA was obtained by incubating the lysate with the magnetic bead-antibody complex overnight at 4 °C. Finally, qRT-PCR was utilized to verify the expression of LINC01977, MEG3 and IgG.First, sufficient cell lysate was obtained from MHCC97H cells. The beads were then eluted and bound by incubation with the corresponding antibodies IgG, RALY (Novus), DHX9 (Novus), RBM39 (Proteintech), mP ≤ 0.05. Volcano map, heatmap, KEGG and GSEA pathway analyses were explored. Each group had three samples. Raw data were deposited in the Gene Expression Omnibus (GEO) database (GSE227449).First, total RNA was acquired from LINC01977 knockdown cells and control cells with TRIzol reagent. The RNA samples were then sequenced using the Illumina HiSeq 4000 platform (Illumina) to generate raw data (LC-Bio). The genes with significant differential expression were chosen based on fold change ≥2.0 and BALB/c male nude mice (6–8 weeks old) were obtained from GemPharmatech Co., Ltd and maintained in specific pathogen-free facilities. Mice were housed at 20 ± 2 °C under a 12-h light/12-h dark photoperiod with lights on at 7:00. All studies involving animals were approved by the Animal Care and Use Committee of the Affiliated Drum Tower Hospital, Medical School of Nanjing University. All researches were in compliance with the Guide for the Care and Use of Laboratory Animals.For the subcutaneous tumor model, stable HCC cell lines were first constructed and inoculated subcutaneously into nude mice. The number of cells per mouse was 5 million, with six mice per group. After 4 weeks, tumors were analyzed by bioluminescence imaging. Subcutaneous tumors of nude mice were then removed, tumor volumes were measured, and tumor weights were weighed. Finally, tumor tissue was prepared into slides for HE, IHC and IF analyses.3. Nude mice were anesthetized, the liver was exposed with scissors and forceps, and a piece of tumor was placed in the liver. After 3 weeks, liver metastases were evaluated by bioluminescence imaging. Livers were then removed and used for HE analysis.For the surgical orthotopic implantation, subcutaneous xenograft tumors were first obtained and then divided into small tissues of ~1 mmFor the lung metastasis model, stable HCC cells were first constructed and then injected into nude mice via the tail vein. The number of cells in each nude mouse was 1 million, and there were six in each group. After 6 weeks, lung metastases were evaluated by bioluminescence imaging. Lungs were then removed and used for HE analysis.For survival analysis, stable HCC cells were constructed first, and then control and LINC01977-overexpressing cells were infused into nude mice. The number of cells in each nude mouse was 1 million, and there were ten in each group. The total observation period was 80 days. Based on the survival time, a survival curve was constructed.Sample sizes were generally chosen based on preliminary data indicating the differences between groups. No statistical method was used to predetermine sample size. Mice were randomly assigned to the groups before treatments, and the investigators were blinded to the allocation of mice into different treatment groups.t test. The chi-square test assessed the correlation between LINC01977 expression and clinicopathological features. Kaplan–Meier curves described the survival function and the log-rank test evaluated the significant differences between different groups. HR and 95% CIs were assessed by univariate and multivariate Cox regression analyses. Each experiment was performed at least three times. Statistical analysis of the data was conducted with SPSS and GraphPad softwares. All P values are *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001.Data are shown as the mean ± s.e.m. Comparisons between two groups were performed using a two-tailed unpaired or paired Student’s supplementary dataOriginal Data File


Hepatocellular carcinoma (HCC) incidence was rising in the United States. Previously, using data collected by the Surveillance, Epidemiology, and End Results (SEER) Program through 2017, we found that overall incidence had begun to decline, although not in Black and American Indian/Alaska Native (AI/AN) populations. Utilizing expanded SEER data encompassing ~50% of the population, we examined secular trends and demographic differences in HCC incidence through 2019.We included cases of HCC diagnosed in adults aged ≥20 years residing in SEER‐22 registry areas. We examined case counts, incidence rates , annual percent changes (APCs), and calendar years when APCs changed significantly.HCC incidence increased from 5.56 in 2000 to 8.89 in 2009 , then rose more slowly during 2009–2015 . After peaking at 10.03 in 2015, incidence fell to 9.20 in 2019 . In Asian/Pacific Islanders (A/PI), the decline began in 2007 and accelerated in 2015 . In 2014, incidence began to fall in the White  and Hispanic populations . In 2016, rates began to fall in Black individuals . In the AI/AN population, incidence was highest in 2017, although the subsequent decline was not statistically significant. In 2019, population‐specific rates were: White, 6.94; Black, 10.74; A/PI, 12.11; AI/AN, 14.56; Hispanic, 15.48.HCC incidence is now decreasing in most US racial/ethnic populations, including among Black individuals. The onset of decline differed among racial/ethnic groups and wide disparities in HCC rates remain. With this larger dataset, we have now examined temporal trends and demographic differences in HCC incidence through 2019 to provide additional information regarding racial disparities for this cancer.22.1https://seer.cancer.gov/about/overview.html). For the present study, we included HCC cases diagnosed between 2000 and 2019 among adults aged ≥20 years residing in 22 SEER registries .The National Cancer Institute funds population‐based SEER Program registries around the United States. For all newly diagnosed cancers among residents of those catchment areas, information is collected on patient demographics, date of diagnosis, and tumor characteristics, but not potential etiologic factors . We defined HCC as cancers with ICD‐O‐3 primary site C220 (liver) and histology codes 8170–8175 (hepatocellular carcinoma). We examined incidence in five nonoverlapping categories for race/ethnicity  All individuals of Hispanic ethnicity were classified as ‘Hispanic’ and excluded from other race/ethnicity categories.2.3We examined the trends in overall HCC incidence and also stratified these analyses by sex, age at diagnosis , race/ethnicity and SEER registry. Rates among American Indian/Alaska Native individuals were restricted to the Purchase/Referred Delivery Areas.We calculated case counts and incidence rates  using SEER*Stat (version 8.3.6), age standardizing the rates to the 2000 US standard population by 5‐year age groups.t‐tests to calculate p‐values.To quantify trends in incidence, we used the Joinpoint Regression Analysis program (version 4.7.0.0) to calculate annual percent changes (APCs) and corresponding 95% confidence intervals (CIs). This program selects the best fitting log‐linear regression model to identify calendar years during which the APCs changed significantly. We used 2‐sided 3During 2000–2019, a total of 194,371 HCC cases were diagnosed among residents of SEER‐22 registry catchment areas aged ≥20 years. HCC rates varied markedly by demographic characteristics  and during 2009–2015  before falling from 2015 to 2019 . For women, HCC incidence began to decline a year earlier than in men , chronic hepatitis C (CHC), alcoholic liver disease (ALD), and nonalcoholic fatty liver disease (NAFLD). Of these factors, ALD and NAFLD are more common, but viral hepatitis infection carries a higher risk for HCC.Major improvements in the treatment of viral hepatitis occurred during 2000–2019. As neither alcohol use nor obesity decreased in the United States during that period,A strength of this study is that SEER employs a rigorous protocol for diagnosis of cancer based on microscopic findings from pathology or cytology, radiography or clinical records and this paper presents results obtained from the largest collection of SEER registries that have been available to date. Our study also has important limitations. SEER does not collect information on etiologic factors that can cause HCC or on foreign nativity. We also lacked the information needed to examine subpopulations within the broadly defined Hispanic and Asian populations.In summary, our study provides welcome evidence of a broad decrease in HCC incidence in the United States, but much remains to be done. Epidemiologically, it will be important to confirm that these trends continue in subsequent years. Efforts to identify and treat individuals with chronic viral hepatitis infections, especially in populations with a high prevalence of infection, as well as those with or at risk for ALD or NAFLD, are essential to ensure that trends in HCC incidence continue to improve.Thomas O'Brien: Conceptualization (lead); formal analysis ; methodology (supporting); writing – original draft (lead); writing – review and editing . Susan S. Devesa: Conceptualization ; formal analysis ; methodology ; writing – review and editing (supporting). Jill Koshiol: Conceptualization (supporting); writing – review and editing (supporting). Jorge Marrero: Conceptualization (supporting); writing – review and editing (supporting). Meredith Shiels: Conceptualization ; formal analysis (lead); methodology (lead); writing – original draft ; writing – review and editing .This work was supported by the Intramural Research Program of the National Institutes of Health, National Cancer Institute, Division of Cancer Epidemiology and Genetics. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. government.No conflict of interest exists for any of the authors.This study was exempt from Institutional Review Board review.


Brain neuroplasticity in which sleep affects the speed of information processing in the elderly population has not been reported. Therefore, this study was conducted to explore the effects of sleep on information processing speed and its central plasticity mechanism in the elderly.t-tests were performed to compare the ALFF, ReHo, and DC maps between the two groups. Then, the relationships among clinical features, fMRI and cognitive function were analyzed using general linear model.A total of 50 individuals aged 60 and older were enrolled in this case control study. All subjects were divided into two groups according to the sleep time: short sleep duration (< 360 min) , and non-short sleep duration (> 360 min) (13 men and 12 women). Resting-state functional magnetic resonance imaging (rs-fMRI) data were collected, and the amplitude of low frequency fluctuation (ALFF), regional homogeneity (ReHo), and degree centrality (DC) were calculated for each participant. Two-sample p < 0.05, AlphaSim correction). The ALFF value of right insula is significantly associated with symbol digit modalities test (SDMT) score .Short sleep duration group showed significantly increased ALFF value in the bilateral middle frontal gyrus and right insula; significantly increased ReHo value in the left superior parietal gyrus, and decreased ReHo value in the right crebellum; significantly decreased DC value in the left inferior occipital gyrus, left superior parietal gyrus and right cerebellum (Short sleep duration and processing speed are significantly associated with remodeling spatial patterns of intrinsic brain activity in the elderly. There iFunctional magnetic resonance imaging (fMRI) is a noninvasive technique that has been widely used to investigate central plasticity or neural plasticity in patients with neurological or psychiatric disorders. This technique provides valuable evidence of the brain’s ability to adapt and reorganize in response to various stimuli or injuries. Since its introduction, fMRI has revolutionized the field of neuroimaging and has become an indispensable tool for researchers and clinicians. By measuring changes in blood flow and oxygenation levels in the brain, fMRI can reveal important insights into the underlying neural mechanisms of various cognitive, emotional, and motor functions. Furthermore, fMRI has the potential to aid in the diagnosis and treatment of neurological and psychiatric disorders by identifying aberrant patterns of neural activity and predicting treatment outcomes . TherefoBrain neuroplasticity in which sleep affects memory function has been well studied, however, brain neuroplasticity in which sleep affects the speed of information processing in the elderly population has not been reported. Disruptions in sleep physiology can lead to age-related cognitive decline . Poor sl2.2.1.A total of 50 individuals aged 60 and older were enrolled in this case control study. The population was homogeneous regarding race  and diet . The study was approved by the local ethics committee at the fourth people’s hospital of Chengdu from April 2021 to March 2022, and all patients were provided with written informed consent according to the Helsinki Declaration. All subjects underwent a neuropsychological assessment and a resting state fMRI (rs-fMRI) scan.The inclusion criteria for all individuals was as follows: (1) Participants aged ≥60 years, (2) Mini-Mental State Examination (MMSE) score  ≥ 24/30,The exclusion criteria for all individuals was as follows: (1) neurological diseases , (2) systemic disease , (3) psychiatric disease , (4) severe hearing or visual impairment, (5) unsuitable for MRI, and (6) < 6 years of education.All subjects were divided into two groups according to the sleep time: short sleep duration (< 360 min) , and non-short sleep duration (> 360 min)  . Sleep d2.2.Two neuropsychologists with >10 years of work experience performed the neuropsychological evaluation; another senior neuropsychologist then reviewed the assessment results. Processing speed was assessed by the Symbol Digit Modalities Test (SDMT) , written2.3.MRI data were acquired using a MAGNETOM Verio 3.0-T scanner  with a 32-channel phased array head coil.3), field of view (FOV) = 230 × 230 mm2, and number of acquisitions = 200.Light and sound shielding were applied before and during the scanning. Subjects were required to close their eyes and keep calm throughout the examination. Their heads were immobilized with foam pads, and their ears were plugged with earplugs. The rs-fMRI data were obtained using a single-pass gradient recalled echoplanar imaging (EPI) sequence with the following parameters: interleaved scanning order, repetition time (TR) = 3,000 ms, slice number = 43, matrix size = 64 × 64, transverse orientation, slice thickness = 3.0 mm, flip angle = 90°, gap = 0 (voxel size = 3.6 × 3.6 × 3.0 mm2.4.http://restfmri.net/forum/RESTplusV1.2), which is based on Statistical Parametric Mapping (SPM12)Functional images of each subject were processed by the Resting-State fMRI Data Analysis Toolkit plus V1.2  was used to conduct our analyses in clinical data. Data are expressed as mean (standard deviation) for continuous variables and percentage for categorical variables. Two-sample p < 0.01 (AlphaSim correction) with a combined individual voxel p < 0.01.The ALFF, ReHo and DC maps were compared between two groups. Two-sample t- test was used for the two groups (two-tailed). AlphaSim correction was used for multiple comparisons, which was performed using the RESTplus toolkit. The resulting statistical map was set at The relationships among clinical features, fMRI and cognitive function were analyzed using general linear model. A general linear model was fitted to assess the independent association between the age/education/body mass index (BMI)/sleep duration and the ALFF/ReHo/DC value (as the dependent variable). Another general linear model was fitted to assess the independent association between the ALFF/ReHo/DC value and SDMT/STT_B score (as the dependent variable).3.3.1.p < 0.05) , MMSE, STT_B and SDMT between the two groups. The sleep duration was 280.80 ± 37.63 in short sleep duration group, and 465.60 ± 58.17 in non-short sleep duration group ( < 0.05) .3.2.p < 0.05, AlphaSim correction, cluster size >21 voxels)  .3.3.p < 0.05, AlphaSim correction, cluster size >55 voxels)  .3.4.Compared to non-short sleep duration group, short sleep duration group showed significantly decreased DC value in the left inferior occipital gyrus, left superior parietal gyrus and right cerebellum  .3.5.β = −0.363, p = 0.033).To investigate the correlation between the age/education/BMI/sleep duration and the ALFF/ReHo/DC value, a general linear model was fitted with the ALFF/ReHo/DC value as the dependent variable and the age/education/BMI/sleep duration as independent variables. 4.Herein, we characterized spatial patterns of intrinsic brain activity and functional connectivity in the elderly with short sleep duration, and explored the relationship between spatial patterns of intrinsic brain activity and information processing speed, Interestingly, we found that short sleep duration and processing speed are significantly associated with remodeling spatial patterns of intrinsic brain activity in the elderly. Our findings point to therapeutic targets for improving processing speed in in the elderly with short sleep duration.ALFF with a typical frequency range (0.01 Hz ~ 0.08 Hz) is considered to describe the local spontaneous neuronal activity, which is thought to be physiologically meaningful . The eldReHo based on Kendall’s coefficient concordance was developed to measure the temporal synchronization of the time series of a given voxel and compare it with its nearest neighbors in a voxel-wise data-driven way . We founIn addition, we found that the elderly with short sleep duration showed significantly decreased DC value in the left inferior occipital gyrus, left superior parietal gyrus and right cerebellum. Lee et al. reported that there are significant characteristic differences in inferior occipital gyrus function between patients with psychogenic insomnia and normal individuals . It was Interestingly, we found the ALFF value of right insula significantly associated with SDMT score, which point outs short sleep duration reduces processing speed via remodeling spatial patterns of intrinsic brain activity and functional connectivity in the elderly. The insula plays an important role in perception, emotion, and self-awareness. Specifically, it has four main functions: 1. Processing and integrating interoceptive information: the insula is a crucial region in the brain that encodes many internal bodily sensations such as heart rate, blood pressure, and digestion. It processes and integrates interoceptive information, helping us better understand the relationship between the body and mind. 2. Generating diverse experiences and behaviors: different insula neurons can respond differently to the same internal signals, which can contribute to generating diverse experiences and behaviors in different contexts. 3. Participating in emotional regulation: the insula is also involved in emotional regulation, influencing emotional experiences and expressions. 4. Participating in the generation of self-awareness: the insula is related to the generation of self-awareness, particularly in relation to bodily self-awareness. These functions are critical to our understanding of the relationship between the body and mind . Age can5.This study also had several limitations. First, the sample size is small, we will expand the sample size to verify the results of this study. Second, we lack objective indicators in evaluating the quality of sleep, we will add sleep monitoring for further in-depth analysis of the relationship between sleep quality and brain function. Third, multimodal fMRI analysis can be performed in subsequent studies, and longitudinal studies also need to be investigated in subsequent phases.6.Short sleep duration and processing speed are significantly associated with remodeling spatial patterns of intrinsic brain activity in the elderly.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by the local ethics committee at the fourth people’s hospital of Chengdu. The patients/participants provided their written informed consent to participate in this study.G-JY and LP: conceptualization and writing–original draft. YZ: methodology. YW and J-LL: validation. X-NZ and XZ: formal analysis. LP: writing, review and editing. All authors contributed to the article and approved the submitted version.This work was supported by Chengdu Municipal Health Commission (no. 2022277).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


S. stercoralis in a cohort of inpatients with invasive bacterial infections of enteric origin to investigate whether the parasite may facilitate these bacterial infections even in the absence of larval hyperproliferation.We assessed the prevalence of S. stercoralis were systematically enrolled over a period of 10 months. S. stercoralis infection was investigated with an in-house PCR on a single stool sample and with at least one serological method (in-house IFAT and/or ELISA Bordier). Univariate, bi-variate and logistic regression analyses were performed.We performed a prospective cross-sectional study in a hospital in northern Italy. Subjects admitted due to invasive bacterial infection of enteric origin and potential previous exposure to Strongyloidiasis was diagnosed in 14/57 patients  of which 10 were Italians  and 4 were migrants . Stool PCR was performed in 43/57 patients (75.4%) and no positive results were obtained.p ≤ 0.05) with male gender, long international travels to areas at higher endemicity, deep extra-intestinal infectious localization and solid tumors. In the logistic regression model, increased risk remained for the variables deep extra-intestinal infectious localization and oncologic malignancy.Strongyloidiasis was found to be significantly associated (Our findings suggest a new role of chronic strongyloidiasis in favoring invasive bacterial infections of enteric origin even in the absence of evident larval dissemination outside the intestinal lumen. Further well-designed studies should be conducted to confirm our results, and possibly establish the underlying mechanisms.The online version contains supplementary material available at 10.1007/s15010-023-02072-1. Strongyloides stercoralis is a nematode (roundworm) transmitted to humans with the passage of filariform larvae  from contaminated soil through intact skin of a host.Its life cycle is very peculiar, as a free-living sexual cycle and an asexual cycle inside the host are present . The parIn high-income countries as Italy, migrants and international travelers from areas at higher endemicity, as well as elderly autochthonous individuals represent the three main categories at risk for the infection –4. AutocS. stercoralis infection involves mainly the immunocompromised population and is defined by either hyperinfection or dissemination, where a great amount of larvae invades host organ systems. The mortality of these forms can reach, respectively, 60% and 100% without treatment [S. stercoralis L3 crossing larvae, but can also escape the gut lumen through parasite-determined intestinal ulcers [Complicated l ulcers , 8.To test the hypothesis that chronic strongyloidiasis may represent a facilitating factor for bacterial extra-intestinal infections of enteric origin even in the absence of larval hyperproliferation, we conducted a prospective cross-sectional study to measure the prevalence of strongyloidiasis in a cohort of patients with severe bacterial extra-intestinal infection whose focus of origin was the gastrointestinal tract  and with epidemiological risk factors for this helminthiasis. The secondary purpose of the study was to identify additional epidemiological and clinical risk factors for strongyloidiasis.We conducted a prospective cross-sectional monocentric study at the ASST Spedali Civili of Brescia University Hospital, affiliated to the University of Brescia  from January to October 2021.Eligible patients were inpatients admitted to the Infectious and Tropical Diseases, Hematology or Internal Medicine divisions due to bacterial extra-intestinal infection whose focus of origin was the gastrointestinal tract, and at least one of the following epidemiological risk criteria for strongyloidiasis: (1) Italians born in 1952 or earlier; (2) international travelers with a life-time stay of more than 14 cumulative days in countries with an estimated prevalence of strongyloidiasis ≥ 5%  age < 18 years; (2) denied or unable to sign the informed consent form; (3) focus of infection different from gastrointestinal tract .S. stercoralis in the study population was not known, we decided to consecutively enroll patients over a pre-defined period of time.Since the background prevalence of S. stercoralis (sensitivity 86.4% (19/22) when larvae are detected with microscopy after concentration by Baermann method; sensitivity 43.8% (14/32) when larvae are not detectable with microscopy after concentration by Baermann method but with coproculture; specificity 100% [S. stercoralis   and/or ELISA Bordier kit  [Enrolled patients provided: (1) a written interviewer-administered questionnaire  on socio-demographic characteristics, exposure and clinical factors. Medical records were consulted to retrieve clinical information; (2) a single stool sample for in-house PCR for  98.3%;) ).Data were entered anonymously into a Microsoft Excel spreadsheet .Strongyloides status. The association between socio-demographic and clinical and behavioral parameters with Strongyloides infection was calculated using Chi2 test. A threshold of significance was defined as α = 0.05, where the null hypothesis is to be rejected when p ≤ 0.05. In tabulations with more than 20% small cell sizes (frequency < 5), Fisher’s exact test was applied. For comparison of age distribution between independent groups as non-normally distributed continuous data, the Wilcoxon rank sum test was applied. The 95% confidence interval (CI) of the overall proportion of Strongyloides-positive cases in the entire dataset was calculated based on the assumption of a binomial distribution. In the analytical part, odds ratios were calculated using a logistic regression model with backward selection of variables by applying the Wald test with a p value cut-off point at 0.5.Data analysis was performed using STATA software, Version SE14 . Variables are presented as proportions, stratified by A total of 57 patients were eligible and all accepted to participate in the study.Of them, 45 (79.0%) were admitted to the Infectious Diseases division, 8 (14.0%) to the Hematology division and 4 (7.0%) to the Internal Medicine division.The most frequent single eligibility criterion for inclusion was being an Italian citizen born in 1952 or earlier , followed by being a migrant  or having traveled to areas with estimated prevalence for strongyloidiasis ≥ 5% for at least 14 cumulative days . Twelve patients (21.1%) had multiple inclusion criteria .Of the enrolled patients, 49/57 (86.0%) were Italian citizens; 3/57 (5.3%) were from other European countries ; 1/57 (1.8%) came from China and 4/57 (7.0%) from Africa . Almost all Italian citizens were born in northern Italy, except one from central Italy and one born in Belgium. All enrolled patients resided in northern Italy at the time of study enrollment. Overall, 40/57 (70.2%) were males; the median age was 76 years (IQR 72–82).E. coli, E. faecalis/faecium and S. gallolyticus  enrolled patients presented an invasive infection by Enterobacteriaceae, whereas 14/57 (24.6%) by other enteric bacteria; most frequently isolated bacteria were Most had a bloodstream infection , while 14/57 (24.6%) presented deep abscesses and/or other deep bacterial localizations , 2/57 (3.5%) had culture-positive meningitis. Only 3/57 (5.3%) had an eosinophil count ≥ 500 cells/µl at the time of admission . One patient (1.8%) died because of hemorrhagic shock due to intestinal bleeding.Chronic or recurrent suggestive symptoms for strongyloidiasis were referred in few cases: pruritus in 7/55 (12.7%), gastrointestinal discomfort in 3/55 (5.5%), respiratory symptoms in 3/55 (5.5%) and skin lesions in 1/55 (1.8%).No one had clinical characteristics suggestive for disseminated strongyloidiasis.Strongyloidiasis was diagnosed in 14/57  patients (positivity of at least one diagnostic technique), of which 10 were Italians  and 4 were migrants  . Stool PCR was negative for all tested patients . IFAT and ELISA were both positive in 2/55 (3.6%) individuals. IFAT alone was positive in 12/55 (21.8%) subjects, while ELISA in 4/57 (7.0%). Further details are present in Table p = 0.04), international travels with duration ≥ 14 continuative days to areas with estimated prevalence ≥ 5% for the parasitosis (p = 0.04), deep extra-intestinal infectious localization (p = 0.01), solid tumors (p = 0.01) and the category “other gastrointestinal disorder” (p = 0.01), that included alcoholic liver cirrhosis, gastric bypass, intestinal occlusion and pseudomelanosis coli. Presence of a hematologic malignancy was inversely correlated with S. stercoralis (p = 0.01). For details, see Tables Strongyloidiasis was found to be significantly associated with male gender p = 0.0, internap ≤ 0.05 for the variables deep extra-intestinal infectious localization, oncologic malignancy and other gastrointestinal disorders  [To the best of our knowledge, this is the first clinical prospective cross-sectional study that investigates the association between ky (USA)  and two ky (USA) , 14 wherThe seroprevalence of strongyloidiasis in our cohort was higher compared with previous studies on non-hospitalized and non-septic individuals , both coRemarkably, our cohort only included three individuals with eosinophilia and all three were negative for strongyloidiasis.Although eosinophilia has usually been considered a key predictive sign for strongyloidiasis, our data suggest that bacterial invasive infections of enteric origin might also be considered a predictive condition. In our Italian cohort, bacterial invasive infection of enteric origin predicted a strongyloidiasis prevalence of 20.4% , not different from the 28% prediction of an eosinophil count > 500 cell/µl in a study of elderly Italians tested with IFAT serology .None of our participants showed a positive result by PCR on stool. This may be explained in part by the known intermittency and low parasite load in stool excreted during chronic strongyloidiasis. As a consequence, even molecular amplification techniques suffer from low sensitivity in these cases , 17, espIn our study, we cannot determine the impact of serological cross reactions leading to potentially false-positive results. However, the serological methods we used have high sensitivity and the existing Italian seroprevalence studies share this limitation.p ≤ 0.05) between strongyloidiasis and the single factor male gender [In the bi-variate analysis, the significant association  in the human gut lumen could alter the composition of the bowel microbiome and the local and/or systemic immune response conferring a higher risk to develop enteric sepsis.We speculate that the mechanism for the favoring role of strongyloidiasis on deep bacterial infections could be a transient crossing of low numbers of larvae through the intestinal barrier with concomitant translocation of enteric bacteria. Otherwise, the long persistence of Our study has limitations, primarily the small study sample. In addition, our patients represent a population consulting a third-level medical facility; hence, external validity of our findings, e.g. towards primary care settings, may be limited as well.The results of our study suggest a new role of chronic strongyloidiasis in favoring extra-intestinal bacterial infections of enteric origin even in the absence of evident larval hyperproliferation and dissemination outside the intestinal lumen. Further well-designed studies should be conducted to confirm our results, eventually establish the underlying mechanisms and the individual and social burden of this phenomenon. In the meantime, both considering the relatively low cost of diagnosis and the harmful potential of undetected strongyloidiasis, we recommend to test for this parasitosis all individuals presenting with invasive bacterial infections of enteric origin if epidemiological risk factors for the parasitosis are present.Supplementary material 1: Written interviewer-administered questionnaire on socio-demographic characteristics, exposure and clinical factors (English version), pdf format. (DOCX 19 KB)Below is the link to the electronic supplementary material.


Understanding related risk factors of health-related quality of life (HRQoL) could avoid treatment failure and provide an insight of personalized treatment approach among people living with HIV/AIDS (PLWH). The objective of this study was to identify factors associated with self-reported treatment qualities and domains of health-related quality of life (HRQoL) among PLWH in Uganda.Data were from “Life on antiretroviral therapy: People’s adaptive coping and adjustment to living with HIV as a chronic condition in Wakiso District, Uganda” in English. The World Health Organization Quality of Life Brief Version (WHOQOL-BREF) questionnaire was used to assess the HRQoL of 263 PLWH in the sample. Considering variance inflation factors, multiple regression analyses were performed to assess the associations between demographic factors, ART obtainment, treatment burden, and self-reported treatment qualities, associations between demographic factors, self-reported treatment qualities, and HRQoL, and association between ART obtainment and HRQoL. Controlling for the confounding effects, several regression anatomies were employed to explore the associations between self-reported treatment qualities and six domains of HRQoL.In the sample, the geographical distribution were urban (5.70%), semi-urban (37.26%), and rural (57.03%). 67.30% of the participants were females. The mean age of the sample was 39.82 years (standard deviation = 9.76) ranging from 22 to 81 years. Multiple logistic regressions reported statistically significant associations of distance to ART facility with self-reported quality of services, advice, manners, and counseling, statistically significant association between self-reported manners quality and four domains of HRQoL, and statistically significant association between TASO membership and domains of HRQoL. Plots from regression anatomies reported that self-reported treatment qualities had statistically significant associations with six domains of HRQoL.Treatment burden, self-reported treatment qualities, ART obtainment, and TASO were possible determinants of individual domains of HRQoL among PLWH in Uganda. PLWH’s HRQoL might be improved by promoting medical quality and optimizing ART obtainment in the healthcare providers’ practice. Findings in this study had important implications for the redesign of clinical guidelines, healthcare delivery, and health care co-ordination among PLWH globally. In order to end HIV/AIDS by 2030, many innovative efforts have been developed to manage antiretroviral therapy (ART), health improvement, and health-related quality of life (HRQoL) among people living with HIV/AIDS (PLWH). Monitoring HRQoL and related risk factors among PLWH could identify treatment fatigue, treatment cancellation, treatment pause, treatment suspension, and therapy postponements in need of extra support and inform a personalized treatment approach. For instance, an international randomized trial discovered immediate treatment could significantly improve self-assessed quality of life (QoL) in healthy PLWH . AnotherRecent studies underscored the psycho-social factors and disease-related factors in the HRQoL among PLWH during ART. For example, sociodemographic factors were strongly associated with lower HRQoL of PLWH , 4. MeanPrior research indicated that having HIV could lead to appreciable treatment and self-management burden. For patients, treatment burden was reflected by adherence to a prescribed, chronic condition self-management regimen , which iA substantial body of studies reported the methods to reduce treatment burden for PLWH –23. As aAs for PLWH, another important concept was treatment quality. The structural aspects of treatment quality could reflect operational quality in healthcare systems. For example, therapeutic relationship, provider characteristics, and treatment approach were identified as major dimensions of treatment quality by patients . AnotherThe literature as mentioned above showed ART treatment burden, self-reported treatment qualities, and ART obtainment had associations with HRQoL, respectively. Adherence to ART was reported to improve HRQoL among PLWH in not Ethiopia , 31 but This study used data from “Life on antiretroviral therapy: People’s adaptive coping and adjustment to living with HIV as a chronic condition in Wakiso District, Uganda” (Project code: RES-062-23-2663) with the questionnaire offered in English and Luganda . The resIn this cross-sectional study, data were collected from November 1, 2010 to August 31, 2013. Three types of ART delivery site in Wakiso District were selected to recruit participants for the study: (1) the HIV clinic at the government hospital in Entebbe; (2) three government health centres (level 3) that have referral links to Entebbe, and (3) the Entebbe branch of a well-established non-governmental organisation, and The AIDS Support Organisation (TASO). A list of eligible patients  was compiled at each facility using patient files. With a systematic random sampling, the 263 PLWH were recruited randomly from the ART delivery sites using patient lists (patients on ART for less than 12 months were excluded). Considering the relationship to head of household, the respondents were head (66.54%), spouse (23.19%), parent (3.80%), sibling (2.66%), child (0.76%), other (2.66%), and not related (0.38%). The rural place (57.03%) was the main place where the respondents resided in, followed by semi-urban place (37.26%) and urban place (5.70%).In this study, the scores of the HRQoL domains were scaled on the basis of Syntax file used to score the WHOQoL-BREF which was adopted on the website of Seattle Quality of Life Group . Two earIn this study, the six domains  of the WHOQoL-BREF questionnaire were considered dependent variables. General QoL was measured by the question: “How do you rate your QoL?” Its response options were very poor (0.76%), poor (4.94%), fair (17.49%), good (57.79%), and very good (19.01%), respectively. Thus, poor general QoL was recorded as 1 (fair and below) and 0 (good or above). General health was measured by the question: “How satisfied with health?” Its response options were very dissatisfied (0.76%), dissatisfied (6.84%), fairly satisfied (9.51%), satisfied (61.22%), and very satisfied (21.67%), respectively. Thus, poor general health was defined as 1 (dissatisfied and below) and 0 (fairly satisfied or above). The other four domains  of WHOQoL-BREF were conversed from Likert scale data into a 0–100 scale.The independent variables considered in this study and their study-specific definitions were under ART obtainment and quality of service. Basically, they were demographic factors , ART obtainment , treatment burden , and self-reported treatment qualities .ART obtainment included number of daily pills, medicine change, ART frequency, and ART duration. Number of daily pills was measured by the question: “How many ART pills do you take per day?” The distribution of response options was 2 (84.03%), 3 (7.22%), 4 (7.98%), 5 (0.38%), and 8 (0.38%). Medicine change was measured by the question: “Have you had to change the medicine and move to a different type of ART?” Its response option of “same medicine as when started” was recoded as 0 and response option of “changed medicine” was recoded as 1. The response option of “don’t know” was treated as a missing value. ART frequency was measured by the question: “How frequently do you visit this ART provider?” The response options were once every week, once every two weeks, once every month, once every two months, and once every three months. Thus, the response options were recoded as 1 when ART frequency ≤ 1 time/month and as 0 when ART frequency > 1 time/month. ART duration was measured by the question: “For how long have you been obtaining ART from this facility?” The answer was calculated by the formula: 12×ART duration years + ART duration months.Here, treatment burden was reflected by the workload of healthcare experienced by PLWH. Thus, according to the questionnaire of “Life on antiretroviral therapy”, the treatment burden was measured by several continuous variables including distance to ART facility, traveling time to ART facility, visiting cost to ART facility, and waiting time at ART facility. Distance to ART facility was measured by the question: “How far do you have to travel to the facility on the day you go to pick up the ART?” Its unit was mile. Travelling time to ART facility was reflected by the question: “How long does it take?” The response was calculated by a formula: 60×travelling hours + travelling minutes. Waiting time at ART facility was measured by the question: “How long do you usually have to wait at the facility on the day you go to pick up your medicine?” The response was calculated by a formula: 60×waiting hours + waiting minutes. Visiting cost to ART facility was measured by the question: “On your last visit how much did you spend on the journey to pick up your medicine?” The response was continuous value with unit of Uganda Shilling.Self-reported services quality, self-reported advice quality, self-reported manners quality, and self-reported counseling quality were measured by the questions: “How would you describe / rate the overall quality of the services that you receive from the facility?”, “How would you rate the medical advice that you receive from the nurse / doctor?”, “How would you rate the manners or personal treatment that you receive from the nurse or doctor who you see at the facility?”, and “How would you describe the quality of the counseling services and advice that you receive from the nurse, doctor or HIV counselor?”, respectively. Their response options were 1 (= poor), 2 (= average/fair/OK), and 3 (= good) and were recoded as “below average” (0 = poor/average/fair/OK) and “above average” (1 = good).p value.Since there were gender differences between HIV burden in terms of the life years lost , descripp value was also used to reflect statistical significance.Several regression analyses were conducted to explore the targeted associations. First of all, multiple logistic regressions were employed to analyze association of demographic factors, ART obtainment, and treatment burden with self-reported treatment qualities. Secondly, multiple regressions were employed to analyze the associations of demographic factors and self-reported treatment qualities with HRQoL were conducted. Since physical QoL, psychological QoL, social QoL, and environment QoL were continuous variables, linear regressions were adopted. Simultaneously, logistic regressions were carried out because general QoL and general health were binary variables. And then, the similar approach was used to explore associations between ART obtainment and HRQoL. Finally, several regression anatomies  were concollin after the regression. As a rule of thumb, variance inflation factor (VIF) values of a group of variables were less than 10 might be unnecessary to merit further investigation. Thus, multicollinearity was not a problem and could be safely ignored.Multicollinearity in this study were diagnosed by stata program p value, respectively. Statistical analyses were done using Stata version 14.0 .The confidence interval and the significance level were denoted by 95% confidence interval (CI) and A total of 263 respondents were studied, with mean age of 39.82 years (standard deviation = 9.76) ranging from 22 to 81 years. Among them, over half were females (67.30%). Considering food (in)security, they ate enough meals (87.07%) or went hungry (12.93%). The response options of ART frequency was once a fortnight (0.76%), once a month (55.89%), once every 2 months (41.44%), and once every 3 months (1.90%). Regarding facility types, they chose Grade A hospital (38.78%), Kasanje HC (III) hospital (10.65%), Nakawuka HC (III) hospital (3.80%), Kigungu (III) hospital (7.22%), TASO hospital (4.18%), and TASO outreach hospital (35.36%). Among the 263 patents, 63.50% respondents changed ART regimen, 36.12% did not changed, and 0.38% did not know whether the ART regimen was changed.Table In Table In Table In Table Controlling for the confounding effects of treatment burden/ART obtainment on the self-reported treatment qualities and their effects on the HRQoL, multiple logistic regression models including all the self-reported treatment qualities variables were constructed based on regression anatomies and plotted by Figs. The findings from this study informed the associations between treatment burden, self-reported treatment qualities, ART obtainment, and HRQoL in Ugandan PLWH. To our best knowledge, it was the first real-world study in Uganda that demonstrated ART frequency and distance to ART facility were associated with self-reported treatment qualities, respectively. The present findings also showed that age, enough meals daily, self-reported advice quality , medicine change, ART frequency (≤ 1 time/month), and ART time (months) were associated with domains of HRQoL, respectively. This study confirmed treatment burden played an important role in self-reported treatment qualities.With unique preferences and HRQoL, PLWH need optimal patient-centered care from health care providers. This was supported by a prior study that quality-of-care satisfaction significantly predicted treatment adherence . SeemingThe associations between demographic factors and ART obtainment was in congruence with a number of early studies. For example, age at diagnosis was directly associated with the type of treatment chosen . Prior sThis study confirmed medicine change played an important role in ART, which was in agreement with previous studies. Medically, Cihlar and Fordyce (2016) reported the key points of regimen selection for combination ART . EvidentRemarkably, treatment burden was found to have association with self-reported treatment qualities in the present study. This could be explained by an early study that distance to HIV care might be associated with retention in care and viral suppression . SimilarThis study also highlighted the role of treatment burden in ART. From a macro perspective, the finding in the present study was in line with several scholars who criticized that the over-burdened health system would not be able to maintain services quality . ClinicaRegarding insignificant association between number of daily pills and domains of HRQoL, a possible explanation for this might result from side effects  due to tHere, TASO had significant associations with part of self-reported treatment qualities and domains of HRQoL. The most plausible reason for the associations has been discussed in previous studies. With respect to policy effectiveness, TASO was confirmed to better PLWH’s QoL , 87. ButRegarding associations between self-reported treatment qualities and six domains of HRQoL, this study encouraged us to analyse international successful experiences and rethink ART system in Uganda. At the nation-level, interventions to improve dietary diversity and food security in Ghana , the US Two strengths of this study were noteworthy. First, this study provided important information on the treatment among PLWH in Uganda, a population that was increasing but has received limited research attention. Second, a broad range of potentially associated treatment factors were explored, which had rarely been considered in the prior literature for ART.This study was limited in the following areas. First, limited by the design of the original questionnaire, nearly all of the variables in this study were patient-centred. Thus, the treatment burden was estimated on the basis of PLWH rather than from both physicians’ and nurses’ perspective. Moreover till now, there was still a need to develop a standard and validated assessment method to measure treatment burden , 103. SeIn conclusion, treatment burden, self-reported treatment qualities, ART obtainment, and TASO membership were possible contributors to change in HRQoL among PLWH in Uganda. Self-reported quality of services, advice, manners, and counseling could be likely to be changed by distance to ART facility. New reform strategies could be enlightened by the statistical outcomes. Thus, it suggested Uganda should reform TASO membership, attenuate ART treatment burden, and optimize ART systems further.


There is a growing interest to support home-based dementia care via contactless monitoring (CM) technologies which do not require any body contact, and allow informal caregivers to remotely monitor the health and safety of people with dementia (PwD). However, sustainable implementation of CM technologies requires a better understanding of informal caregivers' acceptance. This study aimed to examine the (1) general acceptance of CM technology for home-based dementia care, (2) acceptance of different sensor types and use scenarios, and (3) differences between accepters and refusers of CM technology.n = 304 informal caregivers of community-dwelling PwD [Mean(SD) age = 58.5 (10.7)] in the Netherlands and Germany. The survey contained a textual and graphical introduction to CM technologies, as well as questions targeting (1) general acceptance of CM technology, (2) acceptance of seven different contactless sensor types, (3) acceptance of five different use scenarios, and (4) caregivers' own and their care recipients' personal characteristics. Data were examined using descriptive and bivariate analyses.A cross-sectional online survey was conducted among p < .001). RF-based sensors  and light sensors were considered most acceptable, whereas camera-based sensors and audio sensors  were seen as least acceptable for home-based dementia care. Furthermore, participants' acceptance of different use scenarios for CM technology varied significantly (p < .001). The intention to use CM technology was highest for detecting emergencies , and lowest for predicting acute situations . Lastly, accepters and refusers of CM technology significantly differed regarding gender (p = .010), their relation with the PwD (p = .003), eHealth literacy (p = .025), personal innovativeness (p < .001), usage of safety technology (p = .002), and the PwD's type of cognitive impairment (p = .035) and housing situation (p = .023).Participants' general acceptance of CM technology was slightly positive. We found significant differences in acceptability between contactless sensor types (Our findings can inform the development and implementation of acceptable CM technology to support home-based dementia care. Specifically, we show which sensor types and use scenarios should be prioritized from the informal caregiver's view. Additionally, our study highlights several personal characteristics associated with informal caregivers' acceptance of CM technology that should be taken into account during implementation. The worAs the burden on informal caregivers of community-dwelling PwD increases, so too does the need for innovative care solutions, including those from the field of assistive technology . The useHowever, despite their potential, future successful and sustainable implementation of CM technologies in home-based dementia care will, among other things, to a great extent depend on user acceptance . InformaWhen studying user acceptance, different frameworks can be applied. Classical frameworks such as the technology acceptance model  and theHowever, previous research has mainly focused on acceptance of technology in general among PwD and their caregivers –30 or haTaken together, an examination of the technological, contextual, and user-related aspects of acceptance of CM technology for home-based dementia care is needed to allow for better product development and implementation. To our knowledge, this current study is the first to focus on informal caregivers of PwD specifically and to apply a quantitative cross-sectional approach when doing so.1.2.(1)To explore the general acceptance of CM technology for home-based dementia care(2)To explore the acceptance of different sensor types for CM technology(3)To explore the acceptance of different use scenarios for CM technology(4)To explore differences in personal characteristics between accepters and refusers of CM technologyThe overarching aim of this study was to gain more insight into the acceptance of CM technology for home-based dementia care from the informal caregiver's perspective. More specifically, the study aims were:2.2.1.This study applied a quantitative cross-sectional design. Data were collected using an online survey among informal caregivers of community-dwelling PwD in the Netherlands and Germany.2.2.Inclusion criteria for participation were as follows: Participants needed to be (1) 18 years of age or older and (2) voluntarily providing unpaid care to a community-dwelling person with dementia or mild cognitive impairment as main reason of care. Community-dwelling was defined as living at home or in any other non-institutional setting. Participants were acquired by nonprobability convenience sampling. Between May and December of 2022, participants were invited to take part in the study through different channels. Those included (a) mailing lists of caregivers from a previous study who participated in the evaluation of a digital care collaboration platform and indicated their interest in future research participation, (b) the websites of the Dutch and German Alzheimer Society, and c) different peer support groups for informal caregivers on Facebook . Before taking part in the study, participants provided informed consent on the first page of the online survey. Only completed questionnaires were used for further analysis.2.3.n = 3 older informal caregivers  to collect feedback on the clarity of questions and instructions. Adjustments were made accordingly before the survey was set out for data collection. Approval for the study was obtained from the ethics committee of the University of Twente  according to European regulations.The online survey see  was deve2.4.In the following, the different components of the online survey are explained, listed in the same order in which they were presented to participants.2.4.1.(1) -Single items asking for age, gender, relation with care recipient, size of informal care network, geographical distance to care recipient, and care technologies that informal caregivers already make use of -Validated questionnaires measuring perceived caregiver burden [4-item scale of the Zarit Burden Interview (ZBI-4) ], eHealtInformal caregiver characteristics:(2) -Single items asking for age, symptom duration, type of cognitive impairment, housing situation , and usage of professional home care (yes/no).Care recipient characteristics:Informal caregivers of community-dwelling PwD are a diverse group . To obta2.4.2.To give participants an impression of CM technology for home-based dementia care, they were given a short textual introduction together with two illustrations depicting (a) the general concept of CM technology, and (b) a mock-up of a digital platform interface that visualizes possible monitoring information shared within the care network see .2.4.3.We operationalized acceptance of different sensor types as follows: Participants were asked to what extent they consider the use of different sensor types as acceptable in the care of their loved one with dementia, on a Likert scale from 1 (very unacceptable) to 5 (very acceptable). Based on recent reviews on sensor devices for human activity recognition , 12, 41,2.4.4.(1)Detection of emergency situations (2)Prediction of acute situations (3)Monitoring of selfcare behaviors (4)Monitoring of nocturnal wellbeing (5)Monitoring of gradual health status changes Different aspects of acceptance towards each of the scenarios were operationalized and measured as follows: (a) Intention to use [1 item: “I would like to use contactless monitoring technology for (scenario X) in the near future.”], (b) Acceptability for informal caregiver [1 item: “Contactless monitoring technology for (scenario X) is something that I would find acceptable.”], (c) Acceptability for care recipient [1 item: “Contactless monitoring technology for (scenario X) is something that my loved one would find acceptable.”], and (d) Perceived usefulness . For all variables, participants answered on a Likert scale from 1  to 5 .Participants were presented with short descriptions of five different use scenarios for CM technology in home-based dementia care. Those represent the most relevant scenarios that have been distilled from our previous qualitative studies among key stakeholders for CM technology in home-based dementia care , 34:(1)2.4.5.The final part of the survey assessed participants' general acceptance of CM technology, which was operationalized as follows: We measured intention to use CM technology by asking to what extent participants would agree to use CM technology in the care of their loved one with dementia for (a) “At this point in my life” and (b) “When cognitive or physical health of my care recipient declines”. Participants answered on a Likert scale from 1  to 5 .2.5.N, %) were generated for all categorical personal characteristics.The data were analyzed using IBM SPSS statistical software . Descriptive statistics (means and standard deviations) were computed for (a) general acceptance of CM technology, (b) acceptance of different sensor types, (c) acceptance of different use scenarios , and (d) all continuous personal characteristics. Frequency distributions  between use scenarios were tested.U-tests for continuous variables  and Chi-square tests for categorical variables . Statistical significance was set at α < .05.To explore differences in personal characteristics between accepters and refusers, two steps were followed. First, we divided the sample into a) accepters, including those who “strongly agree” or “agree” to use CM technology in the care of their loved one at this point in their life , and b) refusers, including those who “strongly disagree”, “disagree”, or are undecided to use CM technology in the care of their loved one at this point . Subsequently, differences in personal characteristics between accepters and refusers of CM technology were tested by Mann–Whitney 3.3.1.n = 1,000 informal caregivers started the survey, of which n = 568 completed all questions. Of those, n = 304 met all inclusion criteria and were included in the analysis. M = 58.5; SD = 10.7). The majority of participants were female , higher educated , and were the adult children of a person with dementia . Most participants cared for a person with Alzheimer's disease or vascular dementia , who lives alone . On average, the sample reported moderate caregiver burden (score 3 out of 5) and high eHealth literacy (score 3.2 out of 4). At the time of data collection, the majority of the sample neither used safety technology  , digital communication technology , nor technology to support memory/day structure  in the care of their loved one with dementia. Most participants did, however, use digital care platforms to support the coordination of their loved one's care .Upon invitation, 3.2.M = 3.7; SD = 1) compared to at this point in their life  .3.3.M = 4; SD = 0.8) and light sensors , whereas least acceptable sensor types encompassed audio sensors  and camera-based sensors .χ2(6) = 272.71, p < .001]. Post-hoc pairwise comparisons showed that RF-based sensors, light sensors, temperature/humidity sensors, energy meters, and object-tagged sensors were all considered significantly more acceptable than audio sensors (p ≤ .001) and camera-based sensors (p < .001). Furthermore, object-tagged sensors were considered significantly less acceptable than RF-based sensors (p = .003) and light sensors (p < .001).Inferential statistics revealed significant differences in acceptability between sensor types [3.4.M = 3.8; SD = 1) for scenario 1 (detection of emergency situations), and lowest  for scenario 2 (prediction of acute situations).χ2(4) = 63.329, p < .001; acceptability for informal caregiver: χ2(4) = 126.442, p < .001; acceptability for care recipient: χ2(4) = 54.185, p < .001; perceived usefulness: χ2(4) = 48.404, p < .001). Post-hoc pairwise comparisons for intention to use showed that participants' intention to use CM technology for scenario 1 (detection of emergency situations) was significantly higher than for most other scenarios, including scenario 3 (monitoring of selfcare behaviors) (p = .001), scenario 2 (prediction of acute situations) (p < .001), and scenario 5  (p = .032). Furthermore, intention to use CM technology for scenario 4  was significantly higher than for scenario 3 (monitoring of selfcare behaviors) (p = .035) and scenario 2 (prediction of acute situations) (p = .008).Inferential statistics revealed significant differences between use scenarios in all tested variables (intention to use: 3.5.n = 153) and refusers (n = 151) of CM technology for home-based dementia care. -p = .010),more often male (-p = .003) and less often an adult child (daughter/son) of the PwD (p = .003),more often a neighbor or friend (-p = .023),more often caregiver of a PwD who lives alone (-p = .035),less often caregiver of a person with mild cognitive impairment (-p = .025) and personal innovativeness (p < .001),scoring higher on eHealth literacy (-p = .002)more often users of safety technology  in the care of their loved one with dementia  and “monitoring of nocturnal wellbeing”  should be given priority during development and implementation of CM technology for home-based dementia care. We found the lowest intention to use CM technology for “prediction of acute situations” such as fall prediction. While there is a growing research direction within human activity recognition focused on algorithmic analytics to predict  risk levels , 19, thiLastly, our study showed that accepters and refusers of CM technology for home-based dementia care significantly differed regarding gender, their relation with the care recipient, eHealth literacy, personal innovativeness, usage of safety technology, and their care recipient's type of cognitive impairment and housing situation. Interestingly, accepters of CM technology turned out to be significantly more often neighbors or friends, and significantly less often adult children of the PwD. This indicates that informal caregivers who do not have a family bond with the PwD might be more likely to accept CM technology. Adult children, on the other side, are often secondary caregivers in cases where the PwD lives together with a partner, which might reduce their need for CM technology. Additionally, privacy issues might make blood-relatives of PwD more reluctant about using CM technology in the home environment. It was also striking that accepters of CM technology were significantly more often users of safety technology  already in the care of their loved one with dementia. Previous research has shown that assistive technologies which are already used can prevent the acceptance of new technologies . However4.2.Our study does not come without limitations. First, participants' responses were based on the provided survey materials and not on actual use of CM technology. This could have made it more difficult for participants to give their opinion about the technology. However, at the same time, it enabled us to focus on a broad range of possible use scenarios and sensor types at once, including novel RF-based sensor types which are still in development. Our previous qualitative research , 34 has Second, by using an online survey, informal caregivers of PwD with limited digital skills might have been less likely to respond. In fact, on average, our sample showed high eHealth literacy and the majority used digital platforms to support the coordination of their loved one's care. This might cause the sample to be less representative for current informal caregivers of community-dwelling PwD, but possibly more representative for potential future users of CM technology.Lastly, due to the explorative character of this study, no multivariate analysis (binary logistic regression) was performed. Therefore, we cannot conclude which participant characteristics together act as predictors of intention to use CM technology.4.3.Based on our study, several directions for future research can be formulated. First, our study has investigated acceptance of different sensor types and use scenarios of CM technology separately, whereas stated preferences towards combinations of those options can be even more insightful. To investigate this, future research could use a discrete choice experiment (DCE) design. In DCEs, participants make choices between different hypothetical “packages” of a product or service which systematically vary in certain attributes . The choMoreover, future research should further investigate the views and needs of PwD towards CM technology. We have done so in our previous qualitative study  which seLastly, our results have been obtained in the context of CM technology for home-based dementia care. Future research could investigate the extent to which our findings are applicable in the context of informal care for other groups of home-dwelling individuals with chronic conditions.5.Our study shows that Dutch and German informal caregivers of community-dwelling PwD are overall receptive to using CM technology at this point in their life and even moreso at the thought of the cognitive or physical health of their loved declining in the future. In addition, our study provides insight into which sensor types, use scenarios and personal characteristics of informal caregivers should be given more attention to during the development and implementation of CM technology for home-based dementia care. Based on our results, RF-based sensors, light sensors, temperature/humidity sensors, energy meters, and object-tagged sensors are most likely to be accepted by informal caregivers. Moreover, use scenarios for CM technology focused on detecting emergency situations and monitoring of nocturnal wellbeing should be prioritized. Lastly, our study showed several personal characteristics to be associated with CM technology acceptance, including informal caregivers’ gender, relation with care recipient, eHealth literacy, personal innovativeness, usage of safety technology, and their care recipients' type of cognitive impairment and housing situation. Overall, these findings can help to make adequate choices during development and implementation of CM technology for home-based dementia care, thereby increasing the chance of achieving a fit between technology, user, and use context.


We also examined whether there were between-arm differences in engraftment of maternal microbes in the neonatal microbiota. In relation to the control arm, vaginal seeding increased mother-to-neonate microbiota transmission and caused compositional changes and a reduction in alpha diversity (Shannon Index) of the skin and stool microbiota. The neonatal skin and stool microbiota alpha diversity when maternal vaginal microbiota is provided is intriguing and highlights the need of larger randomized studies to determine the ecological mechanisms and effects of vaginal seeding on clinical outcomes.Children delivered by elective, prelabor Cesarean section (C-section) are not exposed to the birth canal microbiota and, in relation to vaginally delivered children, show altered microbiota development. Perturbed microbial colonization during critical early-life windows of development alters metabolic and immune programming and is associated with an increased risk of immune and metabolic diseases. In nonrandomized studies, vaginal seeding of C-section-born neonates partially restores their microbiota colonization to that of their vaginally delivered counterparts, but without randomization, confounding factors cannot be excluded. In a double-blind, randomized, placebo-controlled trial, we determined the effect of vaginal seeding versus placebo seeding (control arm) on the skin and stool microbiota of elective, prelabor C-section-born neonates ( The microbes that first colonize a neonate at birth play a key role in metabolic programming and immune system development –3. Pertu–Cesarean section (C-section)—31.8% of all births in the United States in 2020 —is often8–Bacteroides . We further assessed engraftment of maternal microbes in the infant microbiota by comparing the proportion of microbes shared between mothers and their children.n = 10) or placebo seeding with sterile saline . The mean age of the mothers was 33.4 years  and the mean maternal prepregnancy body mass index (BMI) was 25.6 kg/m2 . Of the 20 neonates, 60% were male, mean birth weight was 3.21 kg , 18/20 received exclusive breast milk as their first feed, and 12/20 were exclusively breastfed at 7 days. See Twenty mother-child dyads were randomized to either vaginal seeding   and Bifidobacterium   in terms of bacterial DNA load (measured by quantitative PCR [qPCR]) or alpha diversity, when comparing mothers from the vaginal-seeding versus control groups  and 2A.  × 10−9) . Relativ 0.0078) .P = 0.033) but not in the transitional stool (P = 0.21) or day 30 stool (P = 0.32)  .P = 0.047 for skin; P = 0.02 for stool), an effect that persisted in the infant stool microbiota by day 30 (P = 0.01).Compared with the control, vaginal seeding caused a significant reduction in alpha diversity (Shannon index) in the skin at day 1  and trany day 30   and to a slightly lesser degree, the transitional stool microbiota   (Treatment (seeding versus control) explained considerable variance in the beta diversity of the skin microbiota of day 1 neonates (ent arm) .10.1128/mbio.00491-23.3FIG S1FIG S1, PDF file, 0.3 MB.Principal-coordinate Analysis of infant microbiota beta diversity (weighted UniFrac) by treatment group. (A) Transitional stool. (B) Day 30 stool. (C) Day 1 skin. A permutational multivariate analysis of variance (PERMANOVA) test was performed, and statistical results were noted on the top of each panel. Download Copyright © 2023 Mueller et al.2023Mueller et al.https://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Enterobacter, in transitional stool and Clostridium sensu stricto in day 30 stool , causing a decrease in the relative abundance of 5 ASVs in the transitional stool  , and an 30 stool  and D. O 18 ASVs .P = 0.0001) (P = 0.009)   and B. I= 0.009) . By day With respect to composition of maternal microbiota sources, the perinatal vaginal microbiota substantially overlapped with the microbiota in other maternal body sites , most no10.1128/mbio.00491-23.4FIG S2FIG S2, PDF file, 0.07 MB.Shared ASVs between vaginal swabs and maternal stool. Download Copyright © 2023 Mueller et al.2023Mueller et al.https://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Our study is the first double-blind, randomized, placebo-controlled trial to determine whether vaginal seeding causes differential engraftment of maternal bacteria in the skin and stool of neonates. The study has important limitations, such as a small sample size and only 2 time points until 30 days of life. Despite these limitations, we observed significant effects of vaginal seeding on the neonatal microbiota. Vaginal seeding of C-section-delivered neonates increased mother-to-neonate microbial transfer and changed the neonatal microbiota in different body sites, urging further studies to determine whether these changes provide health benefits to the more than 1 million babies annually born by C-section in the United States .Bifidobacterium and Bacteroides spp. -approved study (WCG IRB number 1300043) at the Inova Health System in Northern Virginia under the US Food and Drug Administration (FDA) Investigational New Drug Application (IND) number 18076 (with an IND required by the FDA). We recruited pregnant women who were scheduled for an elective C-section at ≥37 weeks gestation and obtained written informed consent. We applied stringent inclusion and exclusion criteria to ensure the least risk of maternal-to-infant transmission of infection as discussed with the FDA and the exclusion of maternal health conditions associated with dysbiosis of the vaginal microbiome. For full details of inclusion and exclusion criteria, see supplemental data and On the day of the scheduled C-section, a team member blind to treatments inserted a gauze moistened with sterile saline into the mother’s vagina. The gauze was incubated in the vagina for approximately 1 h and then removed and placed into a lidded sterile container prior to the mother receiving perioperative antibiotics  and kept at room temperature. Shortly after the insertion of the vaginal gauze, a control gauze was also made by identical preparation of the gauze  but without insertion into the mother’s vagina. The control gauze was also placed in a separate lidded sterile container and kept at room temperature. A team member not blind to the treatments then randomized the mother to either the vaginal seeding arm or the control arm. Randomization occurred in a 1:1 ratio and was also stratified to the following 3 prepregnancy BMI strata: normal weight, overweight, and obese.In the operating room, after C-section delivery of the infant and prior to the infant having skin-skin contact with its mother, the infant was wiped down with the vaginal seeding gauze or the control gauze by a team member who was blind to the treatments. The gauze was wiped first over the infant’s mouth, followed by the face and the rest of the body in a standardized manner as described previously, typically just after the 1-min Apgar score was taken , 23.The We collected detailed data on mothers, including demographics, medical history, anthropometrics, and medication use, including antibiotics during pregnancy. Data collected for the infants included demographics, method of first feeding and subsequent feedings, medication use, and adverse events. We collected and managed study data using the REDCap electronic data capture tools hosted at the Inova Health System.For this study, maternal samples collected and analyzed were as follows: (i) prenatal maternal stool from after 35 weeks gestation, (ii) vaginal swab collected just prior to the vaginal gauze insertion before randomization, (iii) gauze used for swabbing the neonate, (iv) skin swab from the forearm obtained the day after delivery, and (iv) buccal swab obtained the day after delivery. Infant samples collected and analyzed were as follows: (i) transitional stool sample at day of life 2 to 3, (ii) stool sample around day 30 of life , and (iii) skin swab from the forearm (using a dry sterile swab rolled over forearm skin area of approximately 2 by 2 inches) obtained the day after delivery before the infant’s first bath. We stored all samples at −80°C until analysis.We extracted microbial DNA using the Qiagen DNeasy PowerSoil HTP 96 kit  following the manufacturer’s instructions. For 16S rRNA gene sequencing, we amplified the V4 region of 16S rRNA genes using the 515F/806R primers according to the Earth Microbiome Project protocol . We sequACTCCTACGGGAGGCAGCAG; and 518R, ATTACCGCGGCTGCTGG)  with universal 16S gene primers  . We condAll analyses were conducted by the statistical team in a blind manner. Select members of the statistical team were unblinded only after the final analysis of the data. Descriptive statistics (median with interquartile range and percentages) were presented on demographic and clinical characteristics for mothers and infants according to treatment arm.The bioinformatics team conducted all microbiota analyses in a blind manner, and select members were unblinded only after the final analysis.We quality controlled dereplicated 16S sequences without primers and denoised them into amplicon sequence variants (ASVs) using the R package DADA2 (v1.20.0) using the pooled settings for denoising and chimera removal , 38. We 39–We calculated weighted UniFrac distances to obtain the pairwise beta diversity, which we further evaluated by the adonis2 function using the R package vegan to test the significance with 999 permutations . We perfWe used the R package DivNet to estimate Shannon diversity using ecological network regression models. We employed the function “betta”  from theTo estimate the sources of the infant microbiota at different body sites, we performed bacterial source tracking using FEAST . SourcesWe used ANCOM to search for differentiated bacterial taxa by randomization group . We furtPRJNA954027 and are publicly available as of the date of publication.Sequencing reads were deposited in the Sequence Read Archive (SRA) under 10.1128/mbio.00491-23.2DATA SET S1Data Set S1, PDF file, 0.07 MB.Inclusion and exclusion criteria for the clinical trial. Download Copyright © 2023 Mueller et al.2023Mueller et al.https://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the