Roles of caregiver-child interaction on the association of socioeconomic status with early childhood development: a population-based study in rural China.

OBJECTIVE: Socioeconomic status (SES) has been previously associated with children's early development, health, and nutrition; however, evidence about the potential role of caregiver-child interaction in such associations was limited. This study aimed to explore the effect of caregiver-child interaction on the associations of SES with child developmental outcomes, including early neurodevelopment and social-emotional behavior. METHODS: A cross-sectional survey was conducted among 2078 children aged 0-6 in a rural county that just lifted out of poverty in 2020 in Central China. The Ages & Stages Questionnaires-Chinese version (ASQ-C) and the Social-Emotional (ASQ: SE) questionnaire were used to assess children's early neurodevelopment and social-emotional behavior, respectively. Caregiver-child interaction was evaluated with the Brigance Parent-Child Interactions Scale. Regression-based statistical mediation and moderation effect were conducted with the PROCESS macro of SPSS. RESULTS: Children with low SES had an increased risk of suspected neurodevelopmental delay [OR = 1.92, 95% CI: 1.50, 2.44] and social-emotional developmental delay [OR = 1.31, 95% CI: 1.04, 1.66]. The caregiver-child interaction partially mediated the associations of SES with child developmental outcomes; the proportion of the indirect effect was 14.9% for ASQ-C total score and 32.1% for ASQ: SE score. Moreover, the caregiver-child interaction had a significant moderation effect on the association of SES with ASQ-C total score (P < 0.05). A weaker association was observed in children with high-level caregiver-child interaction than in medium and low ones. Similar moderating effects were found among boys but not girls. CONCLUSION: Caregiver-child interaction plays a vital role in the relationship between SES and child development. Children with low SES households will benefit more in terms of their early development from intervention programs strengthening caregiver-child interaction.

Prevalence, toxin-genotype distribution, and transmission of Clostridium perfringens from the breeding and milking process of dairy farms.

This study aimed to elucidate the distribution, transmission, and cross-contamination of Clostridium perfringens during the breeding and milking process from dairy farms. The prevalence of 22.3% (301/1351) yielded 494 C. perfringens isolates; all isolates were type A, except for one type D, and 69.8% (345/494) of the isolates carried atyp. cpb2 and only 0.6% (3/494) of the isolates carried cons. cpb2. C. perfringens detected throughout the whole process but without type F. 150 isolates were classified into 94 pulsed-field gel electrophoresis (PFGE) genotypes; among them, six clusters contained 34 PFGE genotypes with 58.0% isolates which revealed epidemic correlation and genetic diversity; four PFGE genotypes (PT57, PT9, PT61, and PT8) were the predominant genotypes. The isolates from different farms demonstrated high homology. Our study confirmed that C. perfringens demonstrated broad cross-contamination from nipples and hides of dairy cattle, followed by personnel and tools and air-introduced raw milk during the milking process. In conclusion, raw milk could serve as a medium for the transmission of C. perfringens, which could result in human food poisoning. Monitoring and controlling several points of cross-contamination during the milking process are essential as is implementing stringent hygiene measures to prevent further spread and reduce the risk of C. perfringens infection.

Associations of father absence and limited access to books and toys with early childhood development among children aged 0-6 years in a rural county lifted out of poverty in China.

OBJECTIVES: This study aimed to understand the early development and nurturing care environment of children aged 0-6 years in rural China and to evaluate the sex- and age-specific associations of nurturing care environment with child developmental outcomes. METHODS: A cross-sectional survey involving 2078 children aged 0-6 years was conducted using a stratified cluster sampling strategy. We used face-to-face interviews to collect information on child, family and nurturing care. The Ages & Stages Questionnaires-Chinese version and ASQ: Social-Emotional were applied to assess children's neuro- and social-emotional development, respectively. Lower neurodevelopmental scores indicate an increased risk for neurodevelopmental delay, and higher social-emotional scores are indicative to a risk of social-emotional problems. The multiple linear regression model examined the associations of nurturing care environments with childhood development. RESULTS: Among the investigated children, the average age was (42.9 ± 19.8) months and 55.8% were boys; 67.9% of the children had absent fathers because of labour migration and 54.0% had limited access to books and toys. Overall, boys had a lower total neurodevelopmental score than girls; similar gender patterns were also found in the domains of communication, fine motor, problem-solving and person-social. Concurrent absent fathers and limited access to books and toys were significantly associated with reduced neurodevelopmental scores [ß - 11.44, 95% CI (-18.20, -4.68)] and increased social-emotional developmental scores [ß 5.88, 95%CI (1.35, 10.41)] after controlling for confounding factors. Sex-specific analysis only echoed the results in boys. Additionally, having an absent father and limited access to books and toys was associated with lower neurodevelopmental scores [ß - 14.58, 95%CI (-25.41, -3.75)] in children under 3 years of age and higher social-emotional developmental scores among children aged 3-6 years [ß 10.66, 95%CI (5.09, 16.24)]. CONCLUSIONS: Children, especially boys, with absent fathers due to labour migration have poorer neuro- and social-emotional development. Limited access to books and toys and father absence are linked to the children's developmental delay, especially for those under 3 years of age. Our findings suggest that intervention programs in resource-constrained rural areas are desirable; more importantly, such programs should begin before 3 years of age to achieve a benefit-cost outcome.

Ca2+-Chelation-Induced Fabrication of Multistimuli-Responsive Charged Nanogels from Phospholipid-Polymer Conjugates and Use for Drug/Protein Loading.

Thermoresponsive phospholipid-poly(N-isopropylacrylamide) (PL-PNIPAM) conjugates were synthesized via reversible addition fragmentation chain transfer polymerization mediated by a phospholipid-modified trithiocarbonate. Temperature triggered the micellization of the PL-PNIPAM conjugate to form phosphate group-decorated micelles in the aqueous solution. Driven by the chelation of phospholipids and Ca2+, the PL-PNIPAM conjugate and Ca2+ ions formed size-tunable nanoclusters at a temperature beyond the lower critical solution temperature. To fabricate cross-linked nanogels, NIPAM was copolymerized with N-succinimidyl acrylate (NSA) to obtain the PL-P(NIPAM-co-NSA) conjugate bearing pendent cross-linkable functionalities. Subsequently, the size-controllable nanogels containing disulfide linkages were generated at 37 °C by cross-linking the PL-P(NIPAM-co-NSA)/Ca2+ nanoclusters with cystamine through modulation of Ca2+ concentrations. These negatively charged nanogels demonstrate temperature/pH/reduction triple responsiveness. The nanogels can be efficiently loaded with doxorubicin (DOX) and proteins with various isoelectric points. The DOX-loaded nanogels exhibited a temperature/pH/reduction triple-responsive release profile. The immobilized RNase A, BSA, and GOx retained the protein bioactivity. The release of RNase A-loaded nanogels possesses a temperature-responsive profile. The immobilization of Lys and cytochrome C in nanogels inhibited protein bioactivity. However, the addition of NaCl triggered the recovery of bioactivity. These multistimuli-responsive nanogels can provide a versatile platform applicable in biotechnology and drug/protein delivery.

Tracing Clostridium perfringens strains from beef processing of slaughter house by pulsed-field gel electrophoresis, and the distribution and toxinotype of isolates in Shaanxi province, China.

The purpose of this study was to investigate the distribution and specify the transmission and cross-contamination of Clostridium perfringens (C. perfringens) in the beef slaughtering and butchering process. The prevalence of 21.2% (150/708) yielded 208 isolates of C. perfringens, including 80.8% type A and 19.2% type D, 0.4% (3/708) samples carried both type A and D strains, and 72.5% type D isolates carried both cpe and atyp.cpb2 genes. C. perfringens were identified through the whole slaughtering process but no type F (cpe and cpa isolates) was found. 69 isolates were further analyzed and classified into 28 PFGE genotypes and clade I contained 94.2% isolates and 24 PFGE genotypes, which showed the genetic diversity and epidemic correlation. Our study traced C. perfringens contamination along the handling processes and showed a gradually ascending contamination rate during the whole process, revealing widespread cross-contamination from the feces and hides of slaughtered cattle to the carcass in the slaughtering workshop, so as from tools and personnel to meat of the cutting workshops. Strains from different slaughterhouses (regions) have high homology, and type A is the predominant toxinotype. It is necessary to monitor and control several key points of cross-contamination during slaughtering process to reduce a risk of C. perfringens infection.

Prevalence, toxin-typing and antimicrobial susceptibility of Clostridium perfringens in sheep with different feeding modes from Gansu and Qinghai provinces, China.

OBJECTIVE: The purpose of this study was to determine the prevalence and antimicrobial resistance of Clostridium perfringens from sheep (intensive husbandry) in Gansu and Tibetan sheep (extensive husbandry) in Qinghai, China. METHODS: 400 fecal samples (sheep, n = 320, Tibetan sheep, n = 80) were collected from Gansu and Qinghai for C. perfringens isolation. Toxin genes were detected by PCR, antimicrobial susceptibility testing was carried out by broth microdilution method, and whole genome was sequenced using Illumina HiSeq. RESULTS: 83 strains of C. perfringens (sheep, n = 47; Tibetan sheep, n = 36) were isolated from the samples. 44.5% (37/83) of the isolates were positive for cpb2, while 34.9% (29/83) of the isolates were positive for cna. 95.2% isolates were resistant to sulfonamides, followed by tetracycline (22.9%), ampicillin (14.5%), penicillin (10.8%), doxycycline (4.8%), and amoxicillin (1.2%). The isolates from same source shared similar allelic profile and closer genetic relationship. A total of 14 toxin genes and 11 antimicrobial resistance genes were detected among the sequenced isolates, and 10 sequenced C. perfringens isolates carried multiple (n ≥ 3) antimicrobial-resistance genes. Moreover, oxazolidinone-resistant gene optrA was detected in one isolate from Tibetan sheep, which co-harbored tetA(P), aac(6')-aph(2″), ant(6)-Ib, erm(Q), fexA, tet(44), erm(A) and erm(B). CONCLUSIONS: C. perfringens from sheep and Tibetan sheep shared different prevalence rates and antimicrobial-resistance, and the isolates from same source shared closer genetic relationship.

Methionine-Based pH and Oxidation Dual-Responsive Block Copolymer: Synthesis and Fabrication of Protein Nanogels.

In this paper, we synthesized a block copolymer containing pendent thioether functionalities by reversible addition-fragmentation chain transfer polymerization of a tert-butyloxycarbonyl (Boc)-l-methionine-(2-methacryloylethyl)ester (Boc-METMA) monomer using a poly(ethylene glycol) (PEG)-based chain transfer agent. The deprotection of Boc groups resulted in an oxidation and pH dual-responsive cationic block copolymer PEG-b-P(METMA). The block copolymer PEG-b-P(METMA) possessing protonable amine groups was water-soluble at pH < 6.0 and self-assembled to form spherical micelles at pH > 6.0. In the presence of H2O2, the micelles first became highly swollen with time and completely disassembled at last, demonstrating the H2O2-responsive feature because of the oxidation of hydrophobic thioether to hydrophilic sulfoxide. The anticancer drug curcumin (Cur) was entrapped in the polymeric micelles and the Cur-loaded micelles displayed a H2O2-triggered release profile as well as a pH-dependent release behavior, making PEG-b-P(METMA) micelles promising nanocarriers for reactive oxygen species-responsive drug delivery. Taking advantage of the protonated amine groups, the cationic polyelectrolyte PEG-b-P(METMA) formed polyion complex micelles with glucose oxidase (GOx) through electrostatic interactions at pH 5.8. By cross-linking the cores of PIC micelles with glutaraldehyde, the PIC micelles were fixed to generate stable GOx nanogels under physiological conditions. The GOx nanogels were glucose-responsive and exhibited glucose-dependent H2O2-generation activity in vitro and improved storage and thermal stability of GOx. Cur can be encapsulated in the GOx nanogels, and the Cur-loaded GOx nanogels demonstrate the glucose-responsive release profile. The GOx nanogels displayed high cytotoxicity to 4T1 cells and were effectively internalized by the cells. Therefore, these GOx nanogels have potential applications in the areas of cancer starvation and oxidation therapy.

Porcine circovirus type 2 ORF5 protein induces endoplasmic reticulum stress and unfolded protein response in porcine alveolar macrophages.

Porcine circovirus type 2 (PCV2) is the essential infectious agent causing porcine circovirus-associated disease (PCVD) in pigs and one of the important viruses that severely jeopardize the swine husbandry industry. PCV2 elicits the unfolded protein response (UPR) via activation of the PERK pathway, and its capsid protein (Cap) has also been found to induce UPR with subsequent activation of apoptosis. The open reading frame 5 (ORF5) protein is a recently discovered non-structural protein, and its function in PCV2 pathogenesis remains unknown. The aim of this study was to determine whether the PCV2 ORF5 protein could induce endoplasmic reticulum stress (ERS) and UPR in porcine alveolar macrophages (PAMs). pEGFP-tagged ORF5 protein was transiently overexpressed in PAMs. Transmission electron microscopy (TEM) was employed to examine changes in ER morphology, and quantitative real-time PCR and western blotting analysis were used to measure UPR-related cell signaling alterations. We found that the ORF5 protein triggers swelling and degranulation of the ER and upregulates the expression of ERS markers. Further experiments demonstrated that the PCV2 ORF5 protein induces ERS and UPR via the PERK (RNA-activated protein kinase-like endoplasmic reticulum kinase), ATF6 (activating transcription factor 6) and IRE1 (inositol requiring enzyme 1) signaling pathways. Together with previous studies, we provide new information on the ERS-UPR induced by the PCV2 ORF5 protein.

Low esophageal mucosal blood flow in patients with nutcracker esophagus.

Nutcracker esophagus (NE) is characterized by high-amplitude peristaltic esophageal contractions, and these patients often present with symptoms of "angina-like" or noncardiac chest pain. Tissue ischemia is a known cause of visceral pain, and the goal of our present study was to determine whether esophageal wall blood perfusion (EWBP) is reduced in patients with NE. Fourteen normal subjects (mean age 51 yr, 11 men) and 12 patients (mean age 53 yr, 9 men) with NE and noncardiac chest pain were investigated. The EWBP was measured continuously using a custom-designed laser Doppler probe tethered to a Bravo capsule, which anchored it to the esophageal wall. The baseline EWBP in normal subjects was 651 ± 27 perfusion units. In patients with NE, the baseline EWBP was significantly lower than in the normal subjects (451 ± 32 perfusion units). The EWBP decreased after injection of edrophonium (which increases muscle contractions) and increased following sublingual nitroglycerin administration (which relaxes muscle) in normal subjects, as well as in NE patients. Spontaneous pain events during the recording period were often associated with drops in the EWBP. We propose that low EWBP leads to hypoxia of the esophageal tissue, which may be a mechanism of esophageal pain in patients with NE.

Inhibitory motor neurons of the esophageal myenteric plexus are mechanosensitive.

Mechanosensitivity of enteric neurons has been reported in the small intestine and colon, but not in the esophagus. Our earlier in vivo studies show that mechanical stretch of the esophagus in the axial direction induces neurally mediated relaxation of the lower esophageal sphincter, possibly through mechanosensitive motor neurons. However, this novel notion that the motor neurons are mechanosensitive has not been examined in isolated esophageal myenteric motor neurons. The goal of our present study was to examine the mechanosensitivity of esophageal motor neurons in primary culture and elucidate the underlying molecular mechanisms. Immmunocytochemical analysis revealed that >95% cells were positive for the neuronal marker protein gene product 9.5 and that 66% of these cells costained with protein gene product 9.5 and neuronal nitric oxide (NO) synthase. Hypotonic solution induced an increase in the cytoplasm volume in all cells that was independent of extracellular Ca(2+). Hypotonic solution and mechanical stretch induced cytoplasmic free Ca(2+) signaling in ~65% of neurons in the presence, but not absence, of extracellular Ca(2+). Neurons grown on the elastic membrane responded to mechanical stretch by an increase in neuronal size and Ca(2+) signaling simultaneously. Hypotonic stretch-induced cytoplasmic free Ca(2+) signaling was not affected by extracellular Mg(2+), 5-nitro-2-(3-phenylpropylamino)benzoic acid, and nifedipine but was attenuated by 2-aminoethoxydiphenyl borate, Gd(3+), and Grammostola mechanotoxin 4, blockers of the stretch-activated ion channels. In ~57% of the neurons, hypotonic stretch also induced Ca(2+)-dependent cytoplasmic NO production, which was abolished by Grammostola mechanotoxin 4. These results prove that the esophageal inhibitory motor neurons possess a mechanosensitive property and also provide novel insights into the stretch-activated ion channel-Ca(2+)-NO signaling pathway in these neurons.

Circular and longitudinal muscles shortening indicates sliding patterns during peristalsis and transient lower esophageal sphincter relaxation.

Esophageal axial shortening is caused by longitudinal muscle (LM) contraction, but circular muscle (CM) may also contribute to axial shortening because of its spiral morphology. The goal of our study was to show patterns of contraction of CM and LM layers during peristalsis and transient lower esophageal sphincter (LES) relaxation (TLESR). In rats, esophageal and LES morphology was assessed by histology and immunohistochemistry, and function with the use of piezo-electric crystals and manometry. Electrical stimulation of the vagus nerve was used to induce esophageal contractions. In 18 healthy subjects, manometry and high frequency intraluminal ultrasound imaging during swallow-induced esophageal contractions and TLESR were evaluated. CM and LM thicknesses were measured (40 swallows and 30 TLESRs) as markers of axial shortening, before and at peak contraction, as well as during TLESRs. Animal studies revealed muscular connections between the LM and CM layers of the LES but not in the esophagus. During vagal stimulated esophageal contraction there was relative movement between the LM and CM. Human studies show that LM-to-CM (LM/CM) thickness ratio at baseline was 1. At the peak of swallow-induced contraction LM/CM ratio decreased significantly (<1), whereas the reverse was the case during TLESR (>2). The pattern of contraction of CM and LM suggests sliding of the two muscles. Furthermore, the sliding patterns are in the opposite direction during peristalsis and TLESR.

The mediation and interaction effects of Internet addiction in the association between family functioning and depressive symptoms among adolescents: A four-way decomposition.

INTRODUCTION: Previous studies have demonstrated that both family dysfunction and internet addiction (IA) are associated with a higher risk of adolescent depression. However, no study has yet investigated the mechanisms involved. This study aims to explore the mediation and interaction roles of internet addiction (IA) between family functioning and depressive symptoms among adolescents in rural China. METHODS: A multi-stage, stratified cluster, and random sampling was conducted among 3343 adolescents in rural China from October 27 to November 6, 2020. Depressive symptoms, IA, and family functioning were assessed using the Epidemiologic Studies Depression Scale (CESD), the Internet Addiction Test (IAT), and the Family Adaptation Partnership Growth Affection and Resolve Index (APGAR), respectively. Correlation analysis was performed by binary logistic regression. The study employed a four-way decomposition method to explore the potential mechanisms of family functioning on depressive symptoms. RESULTS: The results indicated that family functioning and IA were associated with adolescents' depressive symptoms. The interaction between family functioning and IA accounted for 74 % of the association between family functioning and depression symptoms, while direct effects accounted for 24 %. The "proportion eliminated" (76.11 %) was substantially larger than "proportion mediated" (7.36 %). LIMITATIONS: The cross-sectional design limited to identify the causal relationship among the variables. CONCLUSIONS: We found that family dysfunction synergizes with IA to contribute to the high risk of adolescent depression. Prioritizing at preventing IA in adolescence could be an effective way to mitigate the adverse effects of family dysfunction on depression.

DRA involvement in linaclotide-stimulated bicarbonate secretion during loss of CFTR function.

Duodenal bicarbonate secretion is critical to epithelial protection, as well as nutrient digestion and absorption, and is impaired in cystic fibrosis (CF). We examined if linaclotide, typically used to treat constipation, may also stimulate duodenal bicarbonate secretion. Bicarbonate secretion was measured in vivo and in vitro using mouse and human duodenum (biopsies and enteroids). Ion transporter localization was identified with confocal microscopy, and de novo analysis of human duodenal single-cell RNA sequencing (scRNA-Seq) data sets was performed. Linaclotide increased bicarbonate secretion in mouse and human duodenum in the absence of cystic fibrosis transmembrane conductance regulator (CFTR) expression (Cftr-knockout mice) or function (CFTRinh-172). Na+/H+ exchanger 3 inhibition contributed to a portion of this response. Linaclotide-stimulated bicarbonate secretion was eliminated by down-regulated in adenoma (DRA, SLC26A3) inhibition during loss of CFTR activity. ScRNA-Seq identified that 70% of villus cells expressed SLC26A3, but not CFTR, mRNA. Loss of CFTR activity and linaclotide increased apical brush border expression of DRA in non-CF and CF differentiated enteroids. These data provide further insights into the action of linaclotide and how DRA may compensate for loss of CFTR in regulating luminal pH. Linaclotide may be a useful therapy for CF individuals with impaired bicarbonate secretion.

Morphological, immunocytochemical, and functional characterization of esophageal enteric neurons in primary culture.

The enteric nervous system of the esophagus plays an important role in its sensory and motor functions. Although the esophagus contains enteric neurons, they have never been isolated and characterized in primary culture. We isolated and cultured enteric neurons of the rat esophagus and determined their morphological appearance, chemical coding for neurotransmitters, and functional characteristics. After primary culture for 2 wk, dendrites and axons appeared in the enteric neurons, which usually have one axon and several dendrites. Although the size of neuronal bodies varied from Dogiel type I to type II, their average size was 39 ± 1.8 µm in length and 23 ± 1.4 µm in width. Immmunocytochemical studies revealed that over 95% of these cells were positively stained for two general neuronal markers, PGP 9.5 or Milli-Mark Fluoro. Chemical coding showed that the neurons were positively stained for choline acetyltransferease (53 ± 6%) or nNOS (66 ± 13%). In functional studies, membrane depolarization and stimulation of several G protein-coupled receptors (GPCRs) induced Ca²âº signaling in the esophageal enteric neurons. The GPCR stimulation was found to induce both intracellular Ca²âº release and extracellular Ca²âº entry. The functional expressions of Ca²âº channels (voltage-gated Ca²âº channels and store-operated channels) and Ca²âº pump (sarcoplasmic reticulum Ca²âº-ATPase) were also demonstrated on these neurons. We have grown, for the first time, esophageal enteric neurons in primary culture, and these contain excitatory and inhibitory neurotransmitters. The functional integrity of GPCRs, Ca²âº channels, and Ca²âº pump in these neurons makes them a useful cell model for further studies.

Anatomical disruption and length-tension dysfunction of anal sphincter complex muscles in women with fecal incontinence.

BACKGROUND: Anal sphincter complex muscles, the internal anal sphincter, external anal sphincter, and puborectalis muscles, play an important role in the anal continence mechanism. Patients with symptoms of fecal incontinence have weak anal sphincter complex muscles; however, their length-tension properties and relationship to anatomical disruption have never been studied. OBJECTIVE: This study aimed to assess the anatomy of the anal sphincter complex muscles with the use of a 3-dimensional ultrasound imaging system and to determine the relationship between the anatomical defects and the length-tension property of external anal sphincter and puborectalis muscles in women with incontinence symptoms and in control subjects. DESIGN: Severity of anal sphincter muscle damage was determined by static and dynamic 3-dimensional ultrasound imaging. The length-tension property was determined by anal and vaginal pressure with the use of custom-designed probes. PATIENTS: Forty-four asymptomatic controls and 24 incontinent patients participated in this study. MAIN OUTCOME MEASURES: The anatomical defects and length-tension dysfunction of anal sphincter complex muscles in patients with fecal incontinence were evaluated. RESULTS: The prevalence of injury to sphincter muscles is significantly greater in the incontinent patients than in the controls. Eighty-five percent of patients but only 9% controls reveal damage to ≥2 of the 3 muscles of the anal sphincter complex. Anal and vaginal squeeze pressures increased with the increase in the probe size (length-tension curve) in the majority of controls. In patients, the increase in anal and vaginal squeeze pressures was either significantly smaller than in controls or it decreased with the increasing probe size (abnormal length-tension). LIMITATIONS: We studied patients with severe symptoms. Whether our findings are applicable to patients with mild to moderate symptoms remains to be determined. CONCLUSIONS: The length-tension property of the external anal sphincter and puborectalis muscles is significantly impaired in incontinent patients. Our findings have therapeutic implications for the treatment of anal incontinence.

Obesogenic sleep patterns among Chinese preschool children: A latent profile and transition analysis of the association sleep patterns and obesity risk.

OBJECTIVE: This paper utilized a person-centered approach to examine whether sleep patterns on school and free days are associated with obesity risk in preschool children aged 3-6 years. METHODS: The cross-sectional analysis included 204 children from the Wuhan Healthy Start Project with valid sleep data in at least four consecutive days gathered via Actigraph GT3X+. Based on three domains of sleep duration, sleep onset, and sleep offset, we used latent profile analysis to identify distinct sleep patterns on school and free days separately. Additionally, we conducted latent transition analysis to explore the probabilities of sleep patterns transitions between school and free days. The multivariate logistic regression model investigated the associations of sleep patterns with overweight/obesity (OWO) (BMI ≥ age- and sex-specific 85th percentile) and abdominal obesity (AO) (WC ≥ age- and sex-specific 75th percentile). RESULTS: Two sleep patterns were identified for school days: "EL-sc" (early-to-sleep/longer-duration) (n = 119; 58.3%) and "LS-sc" (late-to-sleep/shorter-duration) (n = 85; 41.7%). Similarly, "LES-fr" (late-to-sleep/early-to-wake/shorter-duration) (n = 118; 57.8%) and "ELL-fr" (early-to-sleep/late-to-wake/longer-duration) (n = 86; 42.2%) patterns were identified for free days. LTA categorized the participants into four distinct transition groups, i.e., "EL-sc→ELL-fr" (32.9%), "EL-sc→LES-fr" (24.0%), "LS-sc→LES-fr" (33.8%), and "LS-sc→ELL-fr" (9.3%). Compared with the "ELsc→ELL-fr", the "LS-sc→LES-fr" had a higher risk of OWO (AOR 4.76; 95% CI: 1.39-20.33) and AO (AOR, 2.78; 95% CI, 1.21-6.62), respectively. Neither "EL-sc→LES-fr" (AOR, 1.11; 95% CI, 0.14-6.67) nor "LS-sc→ELL-fr" (AOR, 0.74; 95% CI, 0.03-6.14) was significantly associated with OWO. Likewise, no significant association was observed for "EL-sc→LES-fr" (AOR, 0.96; 95% CI, 0.35-2.62) and "LS-sc→ELL-fr" (AOR, 0.56; 95% CI, 0.11-2.18) with AO. CONCLUSIONS: "LS-sc→LES-fr" pattern is significantly associated with an increased risk of general and abdominal obesity, indicating its obesogenic nature. Furthermore, although not statistically associated with obesity outcomes, "LS-sc→ELL-fr" and "EL-sc→LES-fr" patterns exhibit a semi-obesogenic characteristic. In addition, we identified a concerning trend that preschool children are at risk of transitioning to and persisting in sleep patterns characterized by delayed and shorter sleep. These findings underscore the importance of implementing interventions and strategies to address sleep patterns as a crucial step to minimize the risk of obesity.

Critical role of down-regulated in adenoma bicarbonate transporter in linaclotide stimulated intestinal bicarbonate secretion.

Duodenal bicarbonate secretion is critical to epithelial protection, nutrient digestion/absorption and is impaired in cystic fibrosis (CF). We examined if linaclotide, typically used to treat constipation, may also alter duodenal bicarbonate secretion. Bicarbonate secretion was measured in vivo and in vitro using mouse and human duodenum. Ion transporter localization was identified with confocal microscopy and de novo analysis of human duodenal single cell RNA sequencing (sc-RNAseq) was performed. Linaclotide increased bicarbonate secretion in mouse and human duodenum in the absence of CFTR expression or function. Linaclotide-stimulated bicarbonate secretion was eliminated by down-regulated in adenoma (DRA) inhibition, regardless of CFTR activity. Sc-RNAseq identified that 70% of villus cells expressed SLC26A3, but not CFTR, mRNA. Linaclotide increased apical membrane expression of DRA in non-CF and CF differentiated enteroids. These data provide insights into the action of linaclotide and suggest linaclotide may be a useful therapy for CF individuals with impaired bicarbonate secretion.

Esophageal wall blood perfusion during contraction and transient lower esophageal sphincter relaxation in humans.

We recently reported that esophageal contraction reduces esophageal wall perfusion in an animal study. Our aim was to determine esophageal wall blood perfusion (EWBP) during esophageal contraction and transient lower esophageal sphincter relaxations (TLESRs) in humans. We studied 12 healthy volunteers. A custom-designed laser Doppler probe was anchored to the esophageal wall, 4-6 cm above the LES, by use of the Bravo pH system so that the laser light beam stay directed toward the esophageal mucosa. A high-resolution manometry equipped with impedance electrodes recorded esophageal pressures and reflux events. Synchronized pressure, impedance, pH, and EWBP recordings were obtained during dry and wet swallows and following a meal. Stable recordings of laser Doppler EWBP were only recorded when the laser Doppler probe was firmly anchored to the esophageal wall. Esophageal contractions induced by dry and wet swallows resulted in 46 ± 9% and 60 ± 10% reduction in the EWBP, respectively (compared to baseline). Reduction in EWBP was directly related to the amplitude (curvilinear fit) and duration of esophageal contraction. Atropine reduced the esophageal contraction amplitude and decreased the EWBP reduction associated with esophageal contraction. TLESRs were also associated with reduction in the EWBP, albeit of smaller amplitude (29 ± 3%) but longer duration (19 ± 2 s) compared with swallow-induced esophageal contractions. We report 1) an innovative technique to record EWBP for extended time periods in humans and 2) contraction of circular and longitudinal muscle during peristalsis and selective longitudinal muscle contraction during TLESR causes reduction in the EWBP; 3) using our innovative technique, future studies may determine whether esophageal wall ischemia is the cause of esophageal pain/heartburn.

Antireflux action of Nissen fundoplication and stretch-sensitive mechanism of lower esophageal sphincter relaxation.

BACKGROUND & AIMS: Surgical fundoplication is an effective treatment for gastroesophageal reflux disease. One of the proposed mechanisms for its antireflux action is that it reduces lower esophageal sphincter (LES) relaxation. We investigated whether fundoplication works through a stretch-sensitive mechanism of LES relaxation. METHODS: Studies were performed in rats. Intravenous and arterial lines were placed and tracheal intubation was performed. A midline laprotomy was performed to place sutures through the esophagus to exert axial stretch on the LES, and the vagus nerve was isolated in the neck for electrical stimulation. The LES pressure was monitored with a 2F solid-state pressure transducer placed through a gastrostomy. Cranial displacement of the LES was recorded using piezoelectric crystals. Data were recorded before and after 360-degree Nissen fundoplication. RESULTS: Axial stretch and vagus nerve stimulation induced cranial displacement of the LES as well as LES relaxation in a dose-dependent manner. LES relaxation and axial stretch were each significantly reduced after fundoplication (P < .01). Nitric-oxide-induced LES relaxation was not affected by fundoplication. Removal of fundoplication restored axial stretch- and vagus nerve-stimulated LES relaxation as well as LES cranial displacement. CONCLUSIONS: Fundoplication reduces LES relaxation by interfering with axial stretch on the LES. Based on this mechanism of the antireflux actions of fundoplication, it might be possible to design new surgical strategies to treat reflux disease and reduce complications of fundoplication surgery.