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Shiga toxin-producing Escherichia coli (STEC) are major foodborne human pathogens that cause mild to hemorrhagic colitis, which could lead to complications of hemolytic uremic syndrome. Seven serogroups, O26, O45, O103, O111, O121, O145, and O157, account for the majority of the STEC illnesses in the United States. Shiga toxins 1 and 2, encoded by stx1 and stx2, respectively, and intimin, encoded by eae gene, are major virulence factors. Cattle are a major reservoir of STEC, but swine also harbor them in the hindgut and shed STEC in the feces. Our objectives were to use a culture method to isolate and identify major and minor serogroups of STEC in finisher pig feces. Shiga toxin genes were subtyped to assess public health implications of STEC. Fecal samples (n = 598) from finisher pigs, collected from 10 pig flows, were enriched in E. coli broth and tested for stx1, stx2, and eae by a multiplex PCR (mPCR) assay. Samples positive for stx1 or stx2 gene were subjected to culture methods, with or without immunomagnetic separation and plating on selective or nonselective media, for isolation and identification of stx-positive isolates. The culture method yielded a total of 178 isolates belonging to 23 serogroups. The three predominant serogroups were O8, O86, and O121. The 178 STEC strains included 26 strains with stx1a and 152 strains with stx2e subtypes. Strains with stx1a, particularly in association with eae (O26 and O103), have the potential to cause severe human infections. All stx2-positive isolates carried the subtype stx2e, a subtype that causes edema disease in swine, but is rarely involved in human infections. Several strains were also positive for genes that encode for enterotoxins, which are involved in neonatal and postweaning diarrhea in swine. In conclusion, our study showed that healthy finisher pigs harbored and shed several serogroups of E. coli carrying virulence genes involved in neonatal diarrhea, postweaning diarrhea, and edema disease, but prevalence of STEC of public health importance was low. HIGHLIGHTS

Pseudocapacitive lithium storage is an effective way to promote the improvement of electrochemical performance for lithium ion batteries. However, the intrinsically sluggish lithium ionic diffusion and the low electronic conductivity of TiO2 limit its capability of pseudocapacitive behavior with fast surface redox reaction. In this work, TiO2 quantum dots confined in 3-dimensional carbon framework have been synthesized by a facile process of reverse microemulsion method combined with heat treatment. The obtained composites effectively combine electrochemical redox with surface pseudocapacitive, showing excellent electrochemical properties. An ultra-high discharge capacity of 370.5 mAh/g can be retained after 200 cycles at a current density of 0.1 A/g. Ultra-long life extends to 10,000 cycles with an average capacity loss of as low as 0.00314% per cycle can be obtained at a high current density of 5.0 A/g, due to the high pesudocapacitance contribution of fast surface redox reaction. Furthermore, the practice application of the obtained electrode is also investigated in a full cell with LiCoO2 as the cathode and a high capacity retention of 93.5% is maintained after 100 cycles at the current density of 0.1 A/g.

Disorders associated with mutations in the Tre2/Bub2/Cdc16 (TBC)1 domain family member 24 gene (TBC1D24) present a wide range of phenotypes, ranging from mild to fatal seizure diseases, non-syndromic deafness, and complex syndromes such as deafness, onychodystrophy, osteodystrophy, and mental retardation(DOOR syndrome). In this study, we introduce three siblings of a previously unreported Chinese family with familial infantile myoclonic epilepsy caused by a homozygous TBC1D24 mutation.Genomic DNA was extracted from whole blood of the proband, his parents, and sisters. TBC1D24 exomes were sequenced by whole exome sequencing then analyzed by genetic analysis with Sanger sequencing validation. The patients were followed up for more than 20 years to summarize their clinical features.Genetic analysis identified a homozygous TBC1D24 mutation (c.241_252del12) in the proband and his sisters. Prediction models suggest that the mutation leads to an alteration in the properties and structure of the TBC1D24 protein, especially in the folding direction of the loop region, which is likely to decrease protein activity. The patients manifested with early-onset myoclonic epilepsy, were prone to status epilepticus, and seizures only occurred during wakefulness. Imaging characteristics included cerebellar atrophy and abnormal cerebellar signals.We report a pedigree case of infantile myoclonic epilepsy caused by a homozygous TBC1D24 mutation. Our long-term clinical follow-up not only enriches the clinical phenotype of the disease, but also provides a clinical experience for the early diagnosis and clinical treatment of the disease.

Shiga toxin-producing E. coli (STEC) are major foodborne pathogens. While many studies have focused on the "top-7 STEC", little is known for minor serogroups. A total of 284 non-top-7 STEC strains isolated from cattle feces were subjected to whole-genome sequencing (WGS) to determine the serotypes, the presence of virulence genes and antimicrobial resistance (AMR) determinants. Nineteen typeable and three non-typeable serotypes with novel O-antigen loci were identified. Twenty-one AMR genes and point mutations in another six genes that conferred resistance to 10 antimicrobial classes were detected, as well as 46 virulence genes. The distribution of 33 virulence genes and 15 AMR determinants exhibited significant differences among serotypes (p 0.05). Among all strains, 81.7% (n = 232) and 14.1% (n = 40) carried stx2 and stx1 only, respectively; only 4.2% (n = 12) carried both. Subtypes stx1a, stx1c, stx2a, stx2c, stx2d, and stx2g were identified. Forty-six strains carried eae and stx2a and therefore had the potential cause severe diseases; 47 strains were genetically related to human clinical strains inferred from a pan-genome phylogenetic tree. We were able to demonstrate the utility of WGS as a surveillance tool to characterize the novel serotypes, as well as AMR and virulence profiles of uncommon STEC that could potentially cause human illness.

Shiga toxin-producing Escherichia coli (STEC) serogroups O26, O45, O103, O111, O121, and O145, called non-O157 STEC, are important foodborne pathogens. Cattle, a major reservoir, harbor the organisms in the hindgut and shed them in the feces. Although limited data exist on fecal shedding, concentrations of non-O157 STEC in feces have not been reported. The objectives of our study were (i) to develop and validate two multiplex quantitative PCR (mqPCR) assays, targeting O-antigen genes of O26, O103, and O111 (mqPCR-1) and O45, O121, and O145 (mqPCR-2); (ii) to utilize the two assays, together with a previously developed four-plex qPCR assay (mqPCR-3) targeting the O157 antigen and three virulence genes (stx1, stx2, and eae), to quantify seven serogroups and three virulence genes in cattle feces; and (iii) to compare the three mqPCR assays to a 10-plex conventional PCR (cPCR) targeting seven serogroups and three virulence genes and culture methods to detect seven E. coli serogroups in cattle feces. The two mqPCR assays (1 and 2) were shown to be specific to the target genes, and the detection limits were 4 and 2 log CFU/g of pure culture-spiked fecal samples, before and after enrichment, respectively. A total of 576 fecal samples collected from a feedlot were enriched in E. coli broth and were subjected to quantification (before enrichment) and detection (after enrichment). Of the 576 fecal samples subjected, before enrichment, to three mqPCR assays for quantification, 175 (30.4%) were quantifiable (≥4 log CFU/g) for at least one of the seven serogroups, with O157 being the most common serogroup. The three mqPCR assays detected higher proportions of postenriched fecal samples (P0.01) as positive for one or more serogroups compared with cPCR and culture methods. This is the first study to assess the applicability of qPCR assays to detect and quantify six non-O157 serogroups in cattle feces and to generate data on fecal concentration of the six serogroups.

Shiga toxin-producing Escherichia coli (STEC) of the serogroups O26, O45, O103, O111, O121, and O145, often called non-O157 STEC, are foodborne pathogens. Cattle are asymptomatic reservoirs for STEC; the organisms reside in the hindgut and are shed in the feces, which serve as the source of food product contaminations. Culture-based detection of non-O157 STEC involves an immunomagnetic separation (IMS) step to capture the specific serogroups in complex matrices, such as feces. The IMS procedure is time consuming and labor intensive because of the need to subject each fecal sample to six individual beads. Therefore, our objective was to evaluate whether pooling of IMS beads affects sensitivity of non-O157 STEC detection compared with using individual IMS beads. The evaluation was done by comparing detection of serogroups in feces spiked with pure cultures (experiments 1 and 2) and from feces (n = 384) of naturally shedding cattle (experiment 3). In spiked fecal samples, detection with pools of three, four, six, or seven beads was similar to, or at times higher than, detection with individual IMS beads. In experiment 3, the proportions of fecal samples that tested positive for the six serogroups as detected by individual or pooled beads were similar. Based on noninferiority tests, detection with pooled beads was not substantially inferior to detection with individual beads (P0.05). In conclusion, the pooling of IMS beads is a better option for detection of STEC serogroups in fecal samples compared with individual beads because the procedure saves time and labor and has the prospect of a higher throughput.

Poor cycling performance of SnS2 induced by the initial structural transformation and subsequent volumetric expansion during lithiating process hinders the widespread application of SnS2 in lithium ion batteries despite its high theoretic capacity. A simple approach promoting the performance of SnS2 is proposed here via strategically introducing rGO/g-C3N4 hybrid to form a robust framework in the designed SnS2 composite. Nano-composite of rGO/g-C3N4@SnS2 where hollow SnS2 spheres with ultra-thin walls are combined with rGO/g-C3N4. The prepared rGO/g-C3N4@SnS2 possesses excellent lithium storage and high rate performance with a recovered capacity of 864.93 mAh/g at the current density of 800 mA/g.

The interplay between the structural configuration and the intrinsic semi-conductive property constrains the further enhancement of the electrochemical performance of TiO2 as an anode material for lithium ion batteries. In this study, hierarchical TiO2@C microrods featured with a macrostructure of microrods composed of a substructure of ultrathin TiO2@C nanosheets with efficient conducting network of carbon surface are constructed strategically on the base of the admirable confluence of the structural configuration and the surface modification. The admirable hierarchical structure facilitates the synergic effect between the surface psedocapacitance for high power density and the electrochemical redox capacity for high energy density, which plays an important role in the enhancement of rate performance. The hierarchical TiO2@C microrods exhibit remarkable rate performance and ultra-long cycling ability with a high reversible capacity of 344.2 mA h g−1 close to its theoretic specific capacity in the first cycle at 0.5C. A reversible capacity of 125.6 mA h g−1 at 60 C has been gained in the second cycle with an average capacity-fading loss of only 0.00375% per cycle over 10,000 cycles. The results clearly reveal that the rational confluence of favourable structure and sound surface modification is effective to improve the electrochemical performance of TiO2.

Application of functional imaging to radiotherapy (RT) is a rapidly expanding field with the development of new modalities and techniques. Functional imaging of PET in conjunction with RT provides new avenues towards the clinical application of dose painting - a new RT strategy delivering optimized dose redistribution according to the functional imaging information to further improve tumour control. Two prototypical strategies of dose painting are reviewed: dose painting by contours (DPBC) and dose painting by numbers (DPBN). DPBN set a linear correlation of the boost dose and image intensity of this same voxel while homogeneous dose is given to the subvolume contoured by a threshold created in PET images in DPBC. Both comply with strict organs at risk (OAR) constraints and are alternatives for boosting subvolumes in clinical practice. This review focuses on the rationale, target validation, dose prescription verification and evaluation and recent clinical achievements in the field of integrating PET imaging into RT treatment planning. Further research is necessary in order to investigate unresolved problems in its routine clinical application thoroughly.

Shiga toxin-producing Escherichia coli (STEC) are important human pathogens, and attention to non-O157 serogroups has increased in recent years. Although cattle are normally considered the primary reservoir for STEC, recent illnesses associated with goat contact have indicated that these animals are important potential reservoirs for the organisms. The prevalence of STEC, particularly non-O157 serogroups, in U.S. goats has not been well described. Our objective was to determine the prevalence of six major non-O157 STEC serogroups in the feces of meat goats. Rectal contents from 296 goats were collected postevisceration at a slaughter plant in the southeastern United States over 9 days during a 12-week period from August through October 2012. Samples were enriched in E. coli broth, and DNA was extracted and used as template in an 11-gene multiplex PCR that detected six non-O157 serogroups (O26, O45, O103, O121, O111, and O145) and virulence genes. Samples were considered positive when at least one non-O157 STEC serotype was present with either stx 1 or stx 2. All six non-O157 serogroups were detected by PCR in our samples, and 14.5% of samples were positive for at least one serogroup. Prevalence of O26 was highest, with 6.4% of goat fecal samples positive. The prevalence of O45 was 3.4%, O103 was 4.4%, O111 was 4.1%, O121 was 1.4%, and O145 was 3.0%. Twenty-two (7.4%) of 296 fecal samples had more than one non-O157 serogroup detected in the feces. Two samples had evidence of three non-O157 STEC serogroups. Goats appear to be an important reservoir for non-O157 STEC, and further work to understand the characteristics, epidemiology, and ecology of STEC in these animals is warranted.

Shiga toxin-producing Escherichia coli (STEC), particularly O157, are major food borne pathogens. Non-O157 STEC, particularly O26, O45, O103, O111, O121, and O145, have also been recognized as a major public health concern. Unlike O157, detection procedures for non-O157 have not been fully developed. Our objective was to develop a multiplex PCR to distinguish O157 and the ‘top six’ non-O157 serogroups (O26, O45, O103, O111, O121, and O145) and evaluate the applicability of the multiplex PCR to detect the seven serogroups of E. coli in cattle feces. Published sequences of O-specific antigen coding genes, rfbE (O157) and wzx and wbqE-F (non-O157), were analyzed to design serogroup-specific primers. The specificity of amplifications was confirmed with 138 known STEC strains and the reaction yielded the expected amplicons for each serogroup. In feces spiked with pooled 7 STEC strains, the sensitivity of the detection was 4.1 × 10 5 CFU/g before enrichment and 2.3 × 10 2 after 6 h enrichment in E. coli broth. Additionally, 216 fecal samples from cattle were collected and tested by multiplex PCR and cultural methods. The multiplex PCR revealed a high prevalence of all seven serogroups (178 [O26], 108 [O45], 149 [O103], 30 [O111], 103 [O121], 5 [O145], and 160 [O157]) of 216 samples in fecal samples. Cultural procedures identified 33.1% (53/160) and 35.5% (11/31) of PCR-positive samples for E. coli O157 and non-O157 serogroups, respectively. Samples that were culture-positive were all positive by the multiplex PCR. The multiplex PCR can be used to identify serogroups of putative STEC isolates.

A multiplex PCR procedure that detects six major virulence genes, fliC, stx1, stx2, eae, rfbE, and hlyA, in Escherichia coli O157:H7 was developed. Analyses of the available sequences of the six major virulence genes and the published primers allowed us to develop the six-gene, multiplex PCR protocol that maintained the specificity of each primer pair. The resulting six bands for fliC, stx1, stx2, eae, rfbE, and hlyA were even and distinct with product sizes of 949, 655, 477, 375, 296, and 199 bp, respectively. The procedure was validated with a total of 221 E. coli strains that included 4 ATCC, 84 cattle, and 57 human E. coli O157:H7 strains as well as 76 non-O157 cattle and human E. coli strains. The results of all 221 strains were similar to the results generated by established multiplex PCR methods that involved two separate reactions to detect five virulence genes (stx1, stx2, eae, fliC, and hlyA). Specificity of the O antigen was indicated by amplification of only O157, and not O25, O26, O55, O78, O103, O111, O127, and O145 E. coli serotypes. Sensitivity tests showed that the procedure amplified genes from a fecal sample spiked with a minimum of 104 CFU/g (10 cells/reaction) of E. coli O157. After a 6-h enrichment of E. coli O157-spiked samples, a sensitivity level of 10 CFU/g was achieved.

Bison is becoming a popular meat source for consumers, but very little is known about the bison's status with respect to Escherichia coli O157:H7. We conducted a study to determine the prevalence and identify virulence genes and pulsed-field genetic types of E. coli O157:H7 in bison. Rectal contents and rectoanal mucosal swab (RAMS) samples were collected from a total of 342 bison at slaughter on seven different dates. Isolation of E. coli O157:H7 was by enrichment, immunomagnetic separation, and plating on selective medium, and identification was based on sorbitol fermentation reaction, indole production, and O157 agglutination test. An overall E. coli O157:H7 prevalence of 47.4% was observed. Fecal prevalence across sampling days ranged from 17 to 83%, with an average of 42.1%. The prevalence in the rectoanal mucosal region ranged from 2.2 to 50%, with an average of 19.9%. All E. coli O157:H7 isolates (n = 212) possessed eae, hlyA, fliC, and stx2 genes. The antiterminator Q gene, Q933, was present in 50.7% of fecal and 38% of RAMS isolates, and Q21 was present in 52.1% of fecal and 61.5% of RAMS isolates. The pulsed-field gel electrophoresis analysis of isolates revealed 11 types (95% Dice similarity) and 19 subtypes (100% Dice similarity). Two pulsed-field genetic types accounted for 76.4% of total isolates. Our study suggests that the prevalence of E. coli O157:H7 in rectal contents or on rectal mucosa of bison is variable, but relatively high overall and bison could serve as an important reservoir for human infection.

Boer-sired goat kids (n = 181) from native pastures were transported to outdoor pens in a beef cattle research facility to evaluate the effects of dietary concentrate level and pen configuration on prevalence of Escherichia coli O157 and to determine if horizontal transmission of E. coli O157 occurs between goats and cattle. Goats were blocked into heavy and light BW groups and randomly assigned to 12 pens, with each containing 15 or 16 animals. Treatments were randomly assigned to pens in a 3 × 2 factorial arrangement. Dietary treatments consisted of diets containing 50, 70, or 90% of the total diet (DM basis) as concentrate, with the balance of the diet as alfalfa hay. Goats were housed in pens with concrete floors, and a large concrete block (45 cm × 75 cm × 150 cm) was placed in the center of 6 of the 12 pens, thus providing two different pen configurations. Fecal samples were collected on day 0, 20, 34, 55, 83, and 105, and cultured for E. coli O157. Feces from cattle housed in adjacent pens were also sampled on day 1, 20, and 34. Isolation and identification of E. coli O157 from fecal samples were by selective enrichment in a broth, followed by immunomagnetic separation and plating on cefixime, tellurite, sorbitol MacConkey agar. Sorbitol-negative colonies that were positive for indole production, positive for O157 antigen agglutination, and speciated by API test, were considered as E. coli O157. Pulsed-field gel electrophoresis (PFGE) of chromosomal DNA was performed on all E. coli O157 isolates to characterize the genetic types and to determine if horizontal transfer of strains occurred between cattle and goats. Detection of the attaching and effacing gene (eae), shiga toxin-1 and -2 genes (stx1 and stx2) was done by polymerase chain reaction (PCR). Mean prevalence of E. coli O157 across all treatments and sampling days was 4.6% and no statistical differences (P > 0.10) were observed for the different pen configurations or diet concentrate levels, but prevalence was numerically higher in the 70% concentrate treatment on 4 of the 6 sampling days. The eae and stx2 genes were present in 95% of the isolates, but the stx1 gene was present in only 51% of the isolates. One E. coli O157 isolate from a goat shared a common PFGE subtype with cattle isolates. Results suggest that in goats with low prevalence of E. coli O157 the level of concentrate in the diet did not influence the shedding pattern.

Escherichia coli O157:H7 causes foodborne illness in humans; cattle are considered a primary reservoir for the organism, and transmission is often through contaminated food products or water. The objective of this study was to determine the genetic diversity of E. coli O157:H7 within a single individual bovine fecal sample based on pulsed-field gel electrophoresis (PFGE) typing. Fecal samples (n=601) were collected from dairy and beef cattle at three separate facilities, and E. coli O157:H7 was isolated by enrichment, immunomagnetic separation, and plating on selective medium. The prevalence of E. coli O157:H7 was 46 (7.7%) of 601. From each positive fecal sample, up to 10 putative colonies were tested, and isolates from samples with at least seven positive colonies were subtyped using PFGE and tested for six major virulence genes by multiplex PCR. A total of 254 E. coli O157:H7 isolates from 27 samples met these criteria and were included in PFGE analysis. Fifteen PFGE subtypes (