[Clinical and genetic analysis of a child with Cerebral creatine deficiency syndrome due to variant of SLC6A8 gene].

OBJECTIVE: To explore the clinical features and genetic variant in a child with Cerebral creatine deficiency syndrome (CCDS). METHODS: A child who had presented at the Affiliated Children's Hospital of Fudan University on March 5, 2021 was selected as the study subject. Whole exome sequencing (WES) was carried out for the child, and candidate variant was verified by Sanger sequencing. The level of creatine in the brain was determined by magnetic resonance spectroscopy. RESULTS: The patient, a 1-year-and-10-month male, had presented with developmental delay and epilepsy. Both his mother and grandmother had a history of convulsions. MRS showed reduced cerebral creatine in bilateral basal ganglia and thalamus. The child was found to harbor a hemizygous splicing variant of the SLC6A8 gene, namely c.1767+1_1767+2insA, which may lead to protein truncation. The variant was not found in the public databases. Both his mother and grandmother were heterozygous carriers for the same variant. CONCLUSION: The hemizygous c.1767+1_1767+2insA variant of the SLC6A8 gene probably underlay the CCDS in this child. Discovery of the novel variant has also expanded the mutational spectrum of the SLC6A8 gene.

Astrocytic GABA transporter 1 deficit in novel SLC6A1 variants mediated epilepsy: Connected from protein destabilization to seizures in mice and humans.

OBJECTIVE: Mutations in γ-aminobutyric acid (GABA) transporter 1 (GAT-1)-encoding SLC6A1 have been associated with myoclonic atonic epilepsy and other phenotypes. We determined the patho-mechanisms of the mutant GAT-1, in order to identify treatment targets. METHODS: We conducted whole-exome sequencing of patients with myoclonic atonic epilepsy (MAE) and characterized the seizure phenotypes and EEG patterns. We studied the protein stability and structural changes with homology modeling and machine learning tools. We characterized the function and trafficking of the mutant GAT-1 with 3H radioactive GABA uptake assay and confocal microscopy. We utilized different models including a knockin mouse and human astrocytes derived from induced pluripotent stem cells (iPSCs). We focused on astrocytes because of their direct impact of astrocytic GAT-1 in seizures. RESULTS: We identified four novel SLC6A1 variants associated with MAE and 2 to 4 Hz spike-wave discharges as a common EEG feature. Machine learning tools predicted that the variant proteins are destabilized. The variant protein had reduced expression and reduced GABA uptake due to endoplasmic reticular retention. The consistent observation was made in cortical and thalamic astrocytes from variant-knockin mice and human iPSC-derived astrocytes. The Slc6a+/A288V mouse, representative of MAE, had increased 5-7 Hz spike-wave discharges and absence seizures. INTERPRETATION: SLC6A1 variants in various locations of the protein peptides can cause MAE with similar seizure phenotypes and EEG features. Reduced GABA uptake is due to decreased functional GAT-1, which, in thalamic astrocytes, could result in increased extracellular GABA accumulation and enhanced tonic inhibition, leading to seizures and abnormal EEGs.

Genomic characterization of a novel bacteriophage STP55 revealed its prominent capacity in disrupting the dual-species biofilm formed by Salmonella Typhimurium and Escherichia coli O157: H7 strains.

Salmonella and Escherichia coli are important foodborne pathogens, forming bacterial biofilms that contribute to their virulence, antimicrobial resistance, and survival on surfaces. Broad lytic phages are promising alternatives to conventional technologies for pathogen biocontrol and reducing biofilms. Herein, we isolated and characterized a novel polyvalent phage STP55 that not only lyse some serotypes of Salmonella, but also some E. coli strains. It had a wide range of pH (4-12) and thermal (30-60 °C) tolerances. The latent time was determined to be 10 min in the one-step growth experiment. Morphological observations by transmission electron microscopy and phylogenetic analysis using terminase gene classified STP55 to family Ackermannviridae in the order Caudovirales, with a complex tail structure. The genome was found to comprise 157,708 bp double-stranded DNA, with 44.57% GC content, 207 predicted ORFs and with no genes associated with antibiotic resistance, toxins, lysogeny, and virulence factors. Particularly, phage STP55 was able to inhibit single- and dual-species biofilms formation by S. Typhimurium ATCC 14028 and E. coli O157: H7, with a reduction percentage of 51.0%, 47.8% and 52.8%, respectively. Moreover, more than 65.0%, 72.9% and 46.2% of an established, single- and dual-species biofilms by S. Typhimurium ATCC 14028 and E. coli O157: H7 were removed after 8 h exposure to the phage STP55, respectively. The elimination effect of STP55 on dual-species biofilm formed on lettuce was further observed by SEM. Overall, our results demonstrated that STP55 is a promising antimicrobial against Salmonella and E. coli.

Prenatal diagnosis and intervention improve developmental outcomes and epilepsy prognosis in children with tuberous sclerosis complex.

AIM: To assess whether prenatal diagnosis and early intervention are beneficial for developmental outcomes and epilepsy prognosis in individuals with tuberous sclerosis complex (TSC). METHOD: This retrospective study originated from a single-centre TSC-specific cohort. We enrolled 273 individuals (138 males, 145 females; 2 years-7 years 6 months, mean 4 years 5 months, SD 1 year 6 months) with definitive TSC who completed TSC1/TSC2 genetic testing and were followed up to 2 years of age. The benefits of early attention and intervention were assessed by comparing epilepsy and developmental outcomes between groups with or without a prenatal diagnosis and with or without presymptomatic preventive intervention. RESULTS: The epilepsy occurrence rate was significantly lower in individuals diagnosed prenatally than in individuals diagnosed postnatally (p = 0.027). In individuals diagnosed prenatally, the epilepsy rate in the preventive intervention subgroup was significantly lower than that in the subgroup without preventive intervention (p = 0.008). Significant improvements in cognitive, language, and motor development were observed in individuals diagnosed prenatally compared to individuals diagnosed postnatally and in the preventive intervention subgroup compared to the subgroup without preventive intervention (p < 0.05). INTERPRETATION: Based on this study, we cautiously speculate that early postpartum intervention may reduce the incidence of epilepsy and intractable epilepsy and improve developmental outcomes. Prophylactic intervention with sirolimus and vigabatrin may reduce the incidence of epilepsy. Larger prospective randomized controlled studies are required to support these findings. WHAT THIS PAPER ADDS: Prenatal diagnosis and early intervention may improve developmental outcomes in children with tuberous sclerosis complex (TSC). Prophylactic intervention with sirolimus and vigabatrin may reduce the incidence of epilepsy. Cardiac and/or intracranial lesions combined with genetic testing can be used to diagnose TSC prenatally.

Identification of TSC2 mosaic mutation limited to cortical tuber with TSC targeted sequencing: a case report and literature review.

BACKGROUND: Tuberous sclerosis complex (TSC) is an autosomal dominant disorder, mainly in childhood presents epilepsy due to cortical tubers. TSC1/TSC2 pathogenic variants cannot be detected in regular molecular genetic testing in around 10-15% of TSC patients. METHODS: We analyzed TSC genes in both cortical tuber, blood and skin samples from a pediatric patient with refractory epilepsy. RESULTS: We found no germline mutations by whole-exome sequencing. Well in targeted sequencing of TSC1/2 data, we identified de novo mutations only in cortical tuber: TSC2 NM_000548.5: exon34:c.4183C>T (p.Gln1395*) in 3% of the alleles. No other TSC mutations were found in patient's blood and skin samples and her parents' blood sample. CONCLUSION: Our case report found TSC2 mosaic mutations can be only limited to cortical tuber in patients with TSC.

Sequestosome 1/p62 Protein Is Associated with Autophagic Removal of Excess Hepatic Endoplasmic Reticulum in Mice.

Xenobiotics exposure increases endoplasmic reticulum (ER) proliferation and cytochrome P-450 (CYP) induction to sustain metabolic requirements. Whether autophagy is essential for the removal of excess ER and CYP and whether an autophagy receptor is involved in this process in mammals remains elusive. In this study, we show that autophagy is induced in mouse livers after withdrawal of the hepatic mitogen 1,4-bis[2-(3,5-dichloropyridyloxy)] benzene (TCPOBOP). Although isolated autophagosomes, autolysosomes, and lysosomes from mouse livers after withdrawal of TCPOBOP contained ER proteins, those in control mouse livers did not. Liver-specific Atg5 knockout mice had higher basal hepatic ER content that was further increased and sustained after withdrawal of TCPOBOP compared with wild-type mice. In addition to regulating ER degradation, our results also suggest that autophagy plays a role in regulating the homeostasis of hepatic CYP because blocking autophagy led to increased CYP2B10 accumulation either at the basal level or following TCPOBOP withdrawal. Furthermore, we found that the autophagy receptor protein sequestosome 1 (SQSTM1)/p62 is associated with the ER. After withdrawal of TCPOBOP, p62 knockout mice had increased ER content in the liver compared with wild-type mice. These results suggest that p62 may act as an autophagy receptor for the autophagic removal of excess ER in the mouse liver. Taken together, our results indicate that autophagy is important for the removal of excess ER and hepatic CYP enzymes in mouse livers, a process associated with the autophagy receptor protein p62.

Parkin regulates mitophagy and mitochondrial function to protect against alcohol-induced liver injury and steatosis in mice.

Alcoholic liver disease claims two million lives per year. We previously reported that autophagy protected against alcohol-induced liver injury and steatosis by removing damaged mitochondria. However, the mechanisms for removal of these mitochondria are unknown. Parkin is an evolutionarily conserved E3 ligase that is recruited to damaged mitochondria to initiate ubiquitination of mitochondrial outer membrane proteins and subsequent mitochondrial degradation by mitophagy. In addition to its role in mitophagy, Parkin has been shown to have other roles in maintaining mitochondrial function. We investigated whether Parkin protected against alcohol-induced liver injury and steatosis using wild-type (WT) and Parkin knockout (KO) mice treated with alcohol by the acute-binge and Gao-binge (chronic plus acute-binge) models. We found that Parkin protected against liver injury in both alcohol models, likely because of Parkin's role in maintaining a population of healthy mitochondria. Alcohol caused greater mitochondrial damage and oxidative stress in Parkin KO livers compared with WT livers. After alcohol treatment, Parkin KO mice had severely swollen and damaged mitochondria that lacked cristae, which were not seen in WT mice. Furthermore, Parkin KO mice had decreased mitophagy, ß-oxidation, mitochondrial respiration, and cytochrome c oxidase activity after acute alcohol treatment compared with WT mice. Interestingly, liver mitochondria seemed able to adapt to alcohol treatment, but Parkin KO mouse liver mitochondria had less capacity to adapt to Gao-binge treatment compared with WT mouse liver mitochondria. Overall, our findings indicate that Parkin is an important mediator of protection against alcohol-induced mitochondrial damage, steatosis, and liver injury.

Targeting the nucleic acid oxidative damage repair enzyme MTH1: a promising therapeutic option.

The accumulation of reactive oxygen species (ROS) plays a pivotal role in the development of various diseases, including cancer. Elevated ROS levels cause oxidative stress, resulting in detrimental effects on organisms and enabling tumors to develop adaptive responses. Targeting these enhanced oxidative stress protection mechanisms could offer therapeutic benefits with high specificity, as normal cells exhibit lower dependency on these pathways. MTH1 (mutT homolog 1), a homolog of Escherichia coli's MutT, is crucial in this context. It sanitizes the nucleotide pool, preventing incorporation of oxidized nucleotides, thus safeguarding DNA integrity. This study explores MTH1's potential as a therapeutic target, particularly in cancer treatment, providing insights into its structure, function, and role in disease progression.

A novel phagomagnetic separation-ATP bioluminescence (PhMS-BL) for rapid and sensitive detection of viable Vibrio parahaemolyticus in aquatic product.

Detection of viable Vibrio parahaemolyticus (V. parahaemolyticus) is a major challenge due to its significant risk to food safety and human health. Herein, we developed a phagomagnetic separation-ATP bioluminescence (PhMS-BL) assay based on phage VPHZ6 for rapid and sensitive detection of viable V. parahaemolyticus. Phage as a recognition element was coupled to magnetic beads to capture and enrich V. parahaemolyticus, shortening detection time and improving method sensitivity. The intracellular ATP released by chemical lysis using CTAB was quantified using firefly fluorescein-adenosine triphosphate bioluminescence system to detect viable bacteria. So, PhMS-BL method was able to detect V. parahaemolyticus in a linear range of 2.3 × 102 to 1.3 × 107 CFU mL-1, with a detection limit of 78 CFU mL-1 within 15 min. It is successfully applied to detect V. parahaemolyticus in spiked lake water, lobster tail meat, and clam meat. The developed detection strategy can rapidly and sensitively detect viable V. parahaemolyticus in food matrixes.

An electrochemical biosensor based on phage-encoded protein RBP 41 for rapid and sensitive detection of Salmonella.

Salmonellosis caused by Salmonella contaminated food poses a serious threat to human health. The rapid and accurate detection of Salmonella is critical for preventing foodborne illness outbreaks. In this study, an electrochemical biosensor was developed using a newly identified biorecognition element, RBP 41, which is capable of specifically recognizing and binding to Salmonella. The biosensor was constructed through a layer-by-layer assembly of graphene oxide (GO), gold nanoparticles (GNPs), and RBP 41 on a glassy carbon electrode (GCE), with the GNPs amplifying the detection signal. The established biosensor was able to detect Salmonella in concentrations ranging from 3 to 106 CFU/mL within approximately 30 min by using differential pulse voltammetry (DPV) signal, and the estimated detection limit was to be 0.2984 Log10 CFU/mL. The biosensor demonstrated excellent specificity and was effective in detecting Salmonella in food matrices, such as skim milk and lettuce. Overall, this study highlights the potential of phage tail receptor binding proteins in biosensing and the proposed biosensor as a promising alternative for rapid and sensitive Salmonella detection in various samples.

A magnetic graphene oxide and UiO-66 based homogeneous dual recognition electrochemical aptasensor for accurate and sensitive detection of aflatoxin B1.

Aflatoxin (AFs) contamination is one of the serious food safety issues. Aflatoxin B1 (AFB1) is the most common and toxic aflatoxin, which has been classified as a class 1 carcinogen by the International Agency for Research on Cancer (IARC). It is extremely destructive to liver tissue. Developing a convenient and sensitive detection technique is essential. In this paper, we developed a homogeneous dual recognition strategy based electrochemical aptasensor for accurate and sensitive detection of aflatoxin B1 (AFB1) based on the magnetic graphene oxide (MGO) and UiO-66. The MGO was synthesized for the recognition and magnetic separation of AFB1 from complex samples. UiO-66/ferrocenecarboxylic acid (Fc)/aptamer composites were constructed as both recognition and signal probes. The probes would specifically capture AFB1 enriched by MGO, which enables dual recognition in homogeneous solution, thus further improving the accuracy of AFB1 detection. The electrochemical aptasensor for AFB1 had a linear range from 0.005 to 500 ng mL-1. Additionally, the limit of detection was 1 pg mL-1. It shows a favorable potential for both sensitive and accurate detection of AFB1 in real samples.

Phage-Based Biosensing for Rapid and Specific Detection of Staphylococcus aureus.

Staphylococcus aureus (S. aureus) is a major foodborne pathogen. Rapid and specific detection is crucial for controlling staphylococcal food poisoning. This study reported a Staphylococcus phage named LSA2302 showing great potential for applications in the rapid detection of S. aureus. Its biological characteristics were identified, including growth properties and stability under different pH and temperature conditions. The genomic analysis revealed that the phage has no genes associated with pathogenicity or drug resistance. Then, the phage-functionalized magnetic beads (pMB), serving as a biological recognition element, were integrated with ATP bioluminescence assays to establish a biosensing method for S. aureus detection. The pMB enrichment brought high specificity and a tenfold increase in analytical sensitivity during detection. The whole detection process could be completed within 30 min, with a broad linear range of 1 × 104 to 1 × 108 CFU/mL and a limit of detection (LOD) of 2.43 × 103 CFU/mL. After a 2 h pre-cultivation, this method is capable of detecting bacteria as low as 1 CFU/mL. The recoveries of S. aureus in spiked skim milk and chicken samples were 81.07% to 99.17% and 86.98% to 104.62%, respectively. Our results indicated that phage-based biosensing can contribute to the detection of target pathogens in foods.

Quantum dot-labeled phage-encoded RBP 55 as a fluorescent nanoprobe for sensitive and specific detection of Salmonella in food matrices.

As a commonly pathogenic bacterium, the rapid detection of Salmonella outbreaks and assurance of food safety require a highly efficient detection method. Herein, a novel approach to Salmonella detection using quantum dot-labeled phage-encoded RBP 55 as a fluorescent nanoprobe is reported. RBP 55, a novel phage receptor binding protein (RBP), was identified and characterized from phage STP55. RBP 55 was functionalized onto quantum dots (QDs) to form fluorescent nanoprobes. The assay was based on the combination of immunomagnetic separation and RBP 55-QDs, which formed a sandwich composite structure. The results showed a good linear correlation between the fluorescence values and the concentration of Salmonella (101-107 CFU/mL) with a low detection limit of 2 CFU/mL within 2 h. The method was used to successfully detect Salmonella in spiked food samples. This approach can be used for the simultaneous detection of multiple pathogens by labeling different phage-encoded RBPs using polychromatic QDs in the future.

Magnetic microbead enzyme-linked immunoassay based on phage encoded protein RBP 41-mediated for rapid and sensitive detection of Salmonella in food matrices.

Rapid and sensitive quantitative detection methods are required to monitor and detect Salmonella throughout the food supply chain and early prevention of foodborne disease outbreaks. In this study, a magnetic microbead enzyme-linked immunoassay (MELISA) based on phage receptor binding protein was developed for rapid enrichment and detection of Salmonella in complex food matrices. RBP 41 from phage T102 acted as a species-specific recognition element for Salmonella by exploiting its strong binding capacity to Salmonella surface receptors. RBP 41-MBs were prepared by coupling recombinant RBP 41 with MBs and used to separate and enrich Salmonella cells from spiked food samples. The captured complexes were further integrated with ELISA procedures by HRP-labeled anti-Salmonella antibody for rapid and accurate detection of Salmonella. The whole method took <1.5 h and the detection limit was 10 CFU/mL. Therefore, MELISA was successfully developed for the detection of Salmonella in various spiked food samples (skim milk, lettuce, and chicken breast). The ELISA reaction process of this method was carried out on magnetic beads. It simplified the process of the traditional ELISA method and reduces the reaction time. This study expanded the use of phage-associated proteins and demonstrated the promising prospects for practical applications in the detection of foodborne pathogens.

A novel compound heterozygous mutation of the CLCN7 gene is associated with autosomal recessive osteopetrosis.

Osteopetrosis is a genetic condition of the skeleton characterized by increased bone density caused by osteoclast formation and function defects. Osteopetrosis is inherited in the form of autosomal dominant and autosomal recessive manner. We report autosomal recessive osteopetrosis (ARO; OMIM 611490) in a Chinese case with a history of scarce leukocytosis, vision and hearing loss, frequent seizures, and severe intellectual and motor disability. Whole-exome sequencing (WES) followed by Sanger sequencing revealed novel compound heterozygous mutations in the chloride channel 7 (CLCN7) gene [c.982-1G > C and c.1208G > A (p. Arg403Gln)] in the affected individual, and subsequent familial segregation showed that each parent had transmitted a mutation. Our results confirmed that mutations in the CLCN7 gene caused ARO in a Chinese family. Additionally, our study expanded the clinical and allelic spectrum of the CLCN7 gene and enhanced the applications of WES technology in determining the etiology of prenatal diagnoses in fetuses with ultrasound anomalies.

BCAT1, as a prognostic factor for HCC, can promote the development of liver cancer through activation of the AKT signaling pathway and EMT.

More and more studies have shown that Branched chain amino acid transaminase 1 (BCAT1) is involved in the occurrence and development of a variety of tumors. However, the mechanism of its occurrence and development in hepatocellular carcinoma (HCC) remains unclear. Here, we demonstrated the relationship between BCAT1 and AKT signaling pathway, as well as EMT, and the clinical significance of BCAT1 by using BCAT1 expression in 5 cell lines and 113 liver cancer and non-liver cancer tissue samples. The results showed that the expression of AKT was positively correlated with BCAT1 in HCC tissues, and BCAT1 could promote the progression of HCC cells through the AKT signaling pathway. Clinical analysis and Bioinformatics technology analysis revealed that BCAT1 was correlated with poor prognosis, and BCAT1 expression in the HCC tissues was evidently correlated with tumor number, vascular invasion, Edmondson grade and TNM stage (P < 0.05). In vitro studies showed that BCAT1 increased the invasion and migration of in MHCC-97H cells a d Huh7 cells. By inhibiting the expression of the BCAT1 gene, we detected the corresponding changes in the expression levels of Twist, E-cadherin and Vimentin, confirming that BCAT1 may promote the invasion and migration of HCC cells through epithelial-mesenchymal transformation (EMT). Overall, BCAT1 can activate AKT signaling pathway and EMT to promote the development and metastasis of HCC cells. this study may provide new ideas and directions for cancer diagnosis and treatment.

Identification of CmbHLH Transcription Factor Family and Excavation of CmbHLHs Resistant to Necrotrophic Fungus Alternaria in Chrysanthemum.

Chrysanthemum morifolium Ramat. 'Huaihuang' is a traditional Chinese medicinal plant. However, a black spot disease caused by Alternaria sp., a typical necrotrophic fungus, has a serious damaging influence on the field growth, yield, and quality of the plant. 'Huaiju 2#' being bred from 'Huaihuang', shows resistance to Alternaria sp. bHLH transcription factor has been widely studied because of their functions in growth development, signal transduction, and abiotic stress. However, the function of bHLH in biotic stress has rarely been studied. To characterize the resistance genes, the CmbHLH family was surveyed in 'Huaiju 2#'. On the basis of the transcriptome database of 'Huaiju 2#' after Alternaria sp. inoculation, with the aid of the Chrysanthemum genome database, 71 CmbHLH genes were identified and divided into 17 subfamilies. Most (64.8%) of the CmbHLH proteins were rich in negatively charged amino acids. CmbHLH proteins are generally hydrophilic proteins with a high aliphatic amino acid content. Among the 71 CmbHLH proteins, five CmbHLHs were significantly upregulated by Alternaria sp. infection, and the expression of CmbHLH18 was the most significant. Furthermore, heterologous overexpression of CmbHLH18 could improve the resistance of Arabidopsis thaliana to necrotrophic fungus Alternaria brassicicola by enhancing callose deposition, preventing spores from entering leaves, reducing ROS accumulation, increasing the activities of antioxidant enzymes and defense enzymes, and promoting their gene expression levels. These results indicate that the five CmbHLHs, especially CmbHLH18, may be considered candidate genes for resistance to necrotrophic fungus. These findings not only increase our understanding of the role CmbHLHs play in biotic stress but also provide a basis by using CmbHLHs to breed a new variety of Chrysanthemum with high resistance to necrotrophic fungus.

Rapid detection of foodborne pathogens in diverse foodstuffs by universal electrochemical aptasensor based on UiO-66 and methylene blue composites.

Rapid and sensitive detection of foodborne pathogens in complex environments is essential for food protection. A universal electrochemical aptasensor was fabricated for the detection of three common foodborne pathogens, including Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), and Salmonella typhimurium (S. typhimurium). The aptasensor was developed based on the homogeneous and membrane filtration strategy. Zirconium-based metal-organic framework (UiO-66)/methylene blue (MB)/aptamer composite was designed as a signal amplification and recognition probe. Bacteria were quantitatively detected by the current changes of MB. By simply changing the aptamer, different bacteria could be detected. The detection limits of E. coli, S. aureus and S. typhimurium were 5, 4 and 3 CFU·mL-1, respectively. In humidity and salt environments, the stability of the aptasensor was satisfactory. The aptasensor exhibited satisfactory detection performance in different real samples. This aptasensor has excellent potential for rapid detection of foodborne pathogens in complex environments.

Telemedicine during the COVID-19 epidemic improves outcomes in children with tuberous sclerosis complex: A 1206 visits retrospective cohort study.

AIMS: To evaluate the benefits of telemedicine in children with tuberous sclerosis complex during the COVID-19 pandemic. METHODS: A retrospective cohort study was conducted, comparing telemedicine and in-person visits within the timeframe spanning from June 1, 2021, to June 1, 2022. Disparities in demographics, emergency visits, hospitalizations, adverse effects (AEs) associated with sirolimus, and the incidence of drug-refractory epilepsy (DRE) between telehealth and in-person care were assessed. Additionally, distinctions between audio and video telehealth, as well as varying frequencies of telehealth encounters, were investigated and reported as odds ratios (ORs). RESULTS: A total of 378 patients with 1206 visits were included, of which 137 were telemedicine patients and 241 were in-person patients. The median age was 5.0 years (IQR 2.8-10.0 years). There were 197 males (52.12%), 691 in-person visits (57.30%), and 515 telemedicine visits (42.70%). Children under 12 years old, those farther away from the center, mothers with more than 12 years of education, and children treated with sirolimus were more likely to visit via telemedicine. Telehealth was associated with significantly fewer emergency visits, hospitalizations, AEs of sirolimus, and DRE. With 10 or more visits, the incidence of emergency visits, hospitalization, and DRE was significantly reduced. CONCLUSION: Telemedicine visits are almost as close in number as in-person visits. Younger patients, patients in remote areas, and mothers with higher education levels are more willing to complete telemedicine visits. Telemedicine visits were associated with a significantly lower number of emergency visits, hospitalizations, and AEs of sirolimus. Patients with more than 10 visits per year seemed to have better clinical outcomes.

Characterization of a novel Jerseyvirus phage T102 and its inhibition effect on biofilms of multidrug-resistant Salmonella.

Biofilm, as a complex microbial community, is a serious and major safety concern in the food industry. Interestingly, some phages could effectively disrupt biofilms. This study characterized a novel isolated Salmonella bacteriophage T102, and its ability to control and remove biofilm produced by multidrug-resistant Salmonella. Phage T102 exhibited a broad host range within the Salmonella genus, especially drug-resistant Salmonella. The genome of phage T102 was comprised of 41,941 bp with 49.7% G + C composition, and with no genes associated with antibiotic resistance or virulence factors. The structural protein profile of phage T102 was subjected to SDS-PAGE and UPLC-MS/MS analysis, among them, 34 peptides were consistent with the hypothetical protein sequences annotated in the genome of T102. The biofilm inhibition assay revealed that phage T102 inhibited the formation of 6 h biofilms by two multidrug-resistant S. Typhimurium strains by 43.17 and 32.42%, respectively. 24 h biofilms formed by S. Typhimurium decreased by 54.94 and 53.67%, respectively, after 2 h of exposure to phage T102. Microscopic observation confirmed the inhibition effect of phage T102 on biofilm formation on spiked lettuce. Overall, our results support new research into the application of bacteriophage for biofilm reduction.