N4-acetyldeoxycytosine DNA modification marks euchromatin regions in Arabidopsis thaliana.

BACKGROUND: Direct analogs of chemically modified bases that carry important epigenetic information, such as 5-methylcytosine (m5C)/5-methyldeoxycytosine (5mC), 5-hydroxymethylcytosine (hm5C)/5-hydroxymethyldeoxycytosine (5hmC), and N6-methyladenosine (m6A)/N6-methyldeoxyadenosine (6mA), are detected in both RNA and DNA, respectively. The modified base N4-acetylcytosine (ac4C) is well studied in RNAs, but its presence and epigenetic roles in cellular DNA have not been explored. RESULTS: Here, we demonstrate the existence of N4-acetyldeoxycytosine (4acC) in genomic DNA of Arabidopsis with multiple detection methods. Genome-wide profiling of 4acC modification reveals that 4acC peaks are mostly distributed in euchromatin regions and present in nearly half of the expressed protein-coding genes in Arabidopsis. 4acC is mainly located around transcription start sites and positively correlates with gene expression levels. Imbalance of 5mC does not directly affect 4acC modification. We also characterize the associations of 4acC with 5mC and histone modifications that cooperatively regulate gene expression. Moreover, 4acC is also detected in genomic DNA of rice, maize, mouse, and human by mass spectrometry. CONCLUSIONS: Our findings reveal 4acC as a hitherto unknown DNA modification in higher eukaryotes. We identify potential interactions of this mark with other epigenetic marks in gene expression regulation.

First Report of Blastocystis Infection in Pigs from Large Farms in Xinjiang, China.

Blastocystis is a common intestinal protozoan in humans and various animals worldwide. A few studies have reported the genetic characterization of Blastocystis in pigs in China, but no epidemiological data are available from the Xinjiang Uygur Autonomous Region. In this study, 801 fecal samples were collected from seven scale pig farms in Xinjiang and tested by polymerase chain reaction and sequence analysis of the partial SSU rRNA gene. The average infection rate of Blastocystis was 21.7% (174/801), with 7.1% in preweaning piglets, 10.0% in postweaning piglets, 31.8% in fattening pigs, and 41.9% in sows (χ2  = 104.89; P < 0.01). Blastocystis subtypes ST1 (7/174), ST3 (2/174), and ST5 (165/174) were identified, with subtype ST5 being predominant in each of the pig farms and in each of the age groups. ST3 and ST5 were identified in preweaning piglets, and ST1, ST3, and ST5 were identified in postweaning piglets. In contrast, only the subtype ST5 was observed in fattening pigs and sows. Genetic polymorphisms were observed at the intrasubtype level, including two variations of ST1 (ST1A, ST1B), and seven of ST5 (ST5A to ST5G), by sequence alignment analysis and phylogenetic analysis. More studies are needed to elucidate the transmission and public health significance of Blastocystis in pigs in various areas.

Evaluation of a novel blood microsampling device for clinical trial sample collection and protein biomarker analysis.

Aim: Evaluation of a novel microsampling device for its use in clinical sample collection and biomarker analysis. Methodology: Matching samples were collected from 16 healthy donors (ten females, six males; age 42 ± 20) via K2EDTA touch activated phlebotomy (TAP) device and phlebotomy. The protein profile differences between sampling groups was evaluated using aptamer-based proteomic assay SomaScan and selected ELISA. Conclusion: Somascan signal concordance between phlebotomy- and TAP-generated samples was studied and comparability of protein abundances between these blood sample collection methods was demonstrated. Statistically significant correlation in selected ELISA assays also confirmed the TAP device applicability to the quantitative analysis of protein biomarkers in clinical trials.

Enterocytozoon bieneusi in donkeys from Xinjiang, China: prevalence, molecular characterization and the assessment of zoonotic risk.

BACKGROUND: Enterocytozoon bieneusi, a zoonotic pathogen, has the potential to infect both immunocompromised and immunocompetent humans. It is found in large number of animals; however, not much is known regarding its prevalence in equine animals, particularly donkeys. This is the first molecular epidemiological evaluation of E. bieneusi in 178 free-ranging donkeys from five countrysides; and 502 farmed donkeys from 18 farms in 12 cities of Xinjiang, China by Nested PCR. RESULTS: E. bieneusi was detected in 2.5% (17/680) donkeys, with 2.6% (13/502) in farmed and 2.2% (4/178) in free-ranging ones. Sequence analysis identified eight ITS genotypes, all belonging to zoonotic Groups 1 or 2, including six known genotypes: horse1 (n = 5), D (n = 3), NCD-2 (n = 3), BEB6 (n = 2), BEB4 (n = 1), and NIAI (n = 1); and two new genotypes: XJD1 (n = 1) and XJD2 (n = 1). CONCLUSIONS: This is the first report confirming the presence of E. bieneusi in donkeys in Xinjiang, China, and indicates the possibility of zoonotic transmission of this pathogenic parasite.

Prevalence and molecular characterization of Cryptosporidium spp. in pigs in Xinjiang, China.

Cryptosporidium spp. are distributed ubiquitously worldwide, and pigs are considered as one of the main reservoir hosts. Eight hundred one porcine fecal specimens were collected from seven intensive pig farms in Xinjiang Uygur Autonomous Region, China. Cryptosporidium spp. were screened via PCR amplification of the small ribosomal subunit RNA gene, and 143 specimens (17.9%, 143/801) from all seven farms tested positive for Cryptosporidium spp. Cryptosporidium prevalence in the pigs differed significantly among farms (p < 0.01). The highest Cryptosporidium spp. prevalence in post-weaned pigs was 39.5% (111/281), followed by fattening pigs (23.2%, 30/129), pre-weaned pigs (1.2%, 2/169), and sows (0/222). Significant differences were observed between age groups (p < 0.01). C. suis was the predominantly identified species (62.9%, 90/143), followed by C. scrofarum (35.7%, 51/143), and C. parvum (1.4%, 2/143). Two C. parvum specimens were subtyped by analyzing the 60-kDa glycoprotein (gp60) gene sequences and were identified as IIdA14G1 and IIdA15G1. To our knowledge, this is the first report of C. parvum infection in pigs in China. The identification of three Cryptosporidium species, including zoonotic C. parvum in pigs in Xinjiang raises concern for the health of both swine animals and personnel in the pig industry.

Genotyping and Zoonotic Potential of Enterocytozoon bieneusi in Pigs in Xinjiang, China.

Enterocytozoon bieneusi is an obligate intracellular fungus, infecting various invertebrate and vertebrate hosts, it is common in humans and causes diarrhea in the immunocompromised. In the present study, 801 fecal specimens were collected from pigs on seven large-scale pig farms in Xinjiang, China. Nested polymerase chain reaction (PCR) amplification of the internal transcribed spacer (ITS) gene showed that the overall E. bieneusi infection rate was 48.6% (389/801). The E. bieneusi infection rates differed significantly among the collection sites (20.0-73.0%) (χ2 = 75.720, df = 6, p < 0.01). Post-weaned pigs had the highest infection rate (77.2%, 217/281), followed by fattening pigs (67.4%, 87/129) and pre-weaned suckling pigs (35.5%, 60/169). Adult pigs had the lowest infection rate (11.3%, 25/222). The E. bieneusi infection rates also differed significantly among age groups (χ2 = 246.015, df = 3, p < 0.01). Fifteen genotypes were identified, including 13 known genotypes (CHC, CS-1, CS-4, CS-7, CS-9, D, EbpA, EbpC, EbpD, H, PigEb4, PigEBITS5, and WildBoar8) and two novel genotypes (XJP-II and XJP-III). Among them, six genotypes (CS-4, D, EbpA, EbpC, H, and PigEBITS5) have been reported in humans. Phylogenetic analysis showed that all the genotypes belonged to Group 1 of E. bieneusi. These findings suggest that pigs may play an important role in transmitting E. bieneusi infections to humans.

Detection and genetic characterization of Giardia duodenalis in pigs from large-scale farms in Xinjiang, China.

To study the presence of Giardia duodenalis in Xinjiang, northwest China, we collected 801 fecal specimens from seven large-scale pig farms and screened them using PCR targeting the SSU rRNA gene. Twenty-one (2.6%) of the specimens from five farms were G. duodenalis-positive, with a significant difference in prevalence among different farms (0-8.7%) (p < 0.01). Giardia duodenalis prevalence was highest in fattening pigs (5.4%, 7/129), followed by sows (3.2%, 7/222), post-weaning piglets (1.8%, 5/281), and pre-weaning piglets (1.2%, 2/169), but there was no significant difference in prevalence among the age groups (p > 0.05). Sequence analysis of the SSU rRNA gene revealed that the 21 G. duodenalis strains belonged to three assemblages: A (n = 2), B (n = 16), and E (n = 3). Assemblage B was the predominant assemblage and was widely distributed in all G. duodenalis-positive farms and age groups. All G. duodenalis-positive specimens were further assayed at the ß-giardin (bg), glutamate dehydrogenase (gdh), and triosephosphate isomerase (tpi) genes, and two tpi, four gdh, and two bg sequences were identified. These data indicate that pigs may be a zoonotic risk and can potentially spread G. duodenalis infection from animals to humans.

Association of A-FABP gene polymorphism and mRNA expression with intramuscular fat content (IMF) in Baicheng-You chicken.

This study aims to assess the association of polymorphisms and mRNA expression of adipocyte-type fatty acid-binding protein (A-FABP) with intramuscular fat (IMF) in the breast muscle (BM) and leg muscle (LM) of Baicheng-You chickens (BYCs). A total of 180 chickens, including sixty black Baicheng-You chickens (BBYCs), sixty silky Baicheng-You chickens (SBYCs) and sixty white Baicheng-You chickens (WBYCs), were reared from 1 to 120 day. A polymerase chain reaction-single-strand conformation polymorphism strategy (PCR-SSCP) was used to detect the polymorphism of the A-FABP gene in the first exon, and the C51T silent mutational site was found. The IMF content with the AA genotype was significantly higher than that with the AG genotype (p = 0.0473) in the LM of WBYC. Thus, this site could be taken as a molecular marker in selecting a higher IMF content of LM in WBYC. A-FABP gene mRNA expression in the BM and LM of BYCs was detected, and a significant positive correlation was observed in the LM of WBYC. These findings provide fundamental data that might be useful in further study of the role of the A-FABP gene in IMF content and fatty metabolism in chickens.

Pharmacogenomic testing: aiding in the management of psychotropic therapy for adolescents with autism spectrum disorders.

Adolescents with autism have higher rates of anxiety than the general adolescent population. They often struggle to express psychological symptoms verbally where their symptoms may manifest as withdrawal and agitation. Adolescent patients with autism have higher rates of polypharmacy and high-risk psychiatric medication use (eg, atypical antipsychotics) than other patients with psychiatric illness. Primary care pediatricians are at the front lines of psychiatric management for patients with autism. Yet, they have inadequate access to pediatric psychiatry for complex medication management. Pharmacogenomic testing can provide personalized drug metabolism profiles for a majority of psychotropic medications. Primary care based pharmacogenomic testing for adolescents with autism on one or more psychiatric medications may help individualize and optimize complex medication regimens, while promoting drug safety.

Correlation of the A-FABP Gene Polymorphism and mRNA Expression with Intramuscular Fat Content in Three-Yellow Chicken and Hetian-Black Chicken.

The adipocyte-type fatty acid-binding protein (A-FABP) is considered a candidate gene for fat metabolism; thus, it affects fat deposition in chickens. The present study was designed to examine the polymorphism and mRNA abundance of the A-FABP gene with intramuscular fat (IMF) in the pectoralis muscles (PM) and leg muscles (LM) of Three-yellow Chicken (TYC) and Hetian-black Chicken (HTBC). In total, 60 TYCs and 60 HTBCs were sacrificed using exsanguination at market age. The IMF contents of the PM and LM in the HTBC were significantly higher than those in the TYC. Three genotypes of the A-FABP gene first exon, AA, AB, and BB, were examined by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP), and a C51 T mutational site, which is a silent substitution mutation, was revealed. The IMF contents of the AA genotype in the PM of the HTBC were significantly higher than those in the AB genotype; thus, the C51 T mutable site is a gene marker for selecting a higher IMF content in the PM of the HTBC. The relative expression of the A-FABP mRNA in the LM of the HTBC, which was measured by quantitative real-time PCR, was significantly higher than in the TYC. A significantly positive association was detected between A-FABP expression with the IMF contents of the PM and LM of both the TYC and the HTBC. These results provide basic data that might be helpful to further research the role of the A-FABP gene in fat deposition and fatty acid metabolism in chickens.

Association of H-FABP gene polymorphisms with intramuscular fat content in Three-yellow chickens and Hetian-black chickens.

BACKGROUND: To explore the relationship between the heart-type fatty acid binding protein (H-FABP) gene and intramuscular fat (IMF), a polymorphism of the second exon of the H-FABP gene was investigated in 60 Three-yellow chickens (TYCs) and 60 Hetian-black chickens (HTBCs). RESULTS: The IMF contents of the cardiac, chest and leg muscles in HTBC were increased compared with TYC. Both TYC and HTBC populations exhibited Hardy-Weinberg Equilibrium (HWE) according to the χ(2) test. Three variations of the two birds were detected, namely, G939A, G982A and C1014T. HTBCs with the TT genotypes exhibit increased IMF content in the chest muscles compared with the TC genotype. Thus, the G982A site could be considered a genetic marker for selecting increased IMF content in the chest muscles of HTBC. The correlation coefficients revealed that H-FABP mRNA expression was negatively correlated with the IMF content in the cardiac, chest and leg muscles of HTBC and in the cardiac and chest muscles of TYC. The relative mRNA expression of H-FABP was reduced in the cardiac and leg muscles of HTBC compared with TYC, but this difference was not observed at the protein level, as assessed by Western blot analysis. CONCLUSIONS: These findings offer essential data that can be useful in the breeding program of HTBC and future research exploring the role of H-FABP in IMF deposition and regulation in chickens.

Correlation between Heart-type Fatty Acid-binding Protein Gene Polymorphism and mRNA Expression with Intramuscular Fat in Baicheng-oil Chicken.

This study aims to determine the polymorphism and mRNA expression pattern of the heart-type fatty acid-binding protein (H-FABP) gene and their association with intramuscular fat (IMF) content in the breast and leg muscles of Baicheng oil chicken (BOC). A total of 720 chickens, including 240 black Baicheng oil chicken (BBOC), 240 silky Baicheng oil chicken (SBOC), and 240 white Baicheng oil chicken (WBOC) were raised. Three genotypes of H-FABP gene second extron following AA, AB, and BB were detected by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) strategy. The G939A site created AA genotype and G956A site created BB genotype. The content of IMF in AA genotype in breast muscle of BBOC was significantly higher than that of AB (p = 0.0176) and the genotype in leg muscle of WBOC was significantly higher than that of AB (p = 0.0145). The G939A site could be taken as genetic marker for higher IMF content selecting for breast muscle of BBOC and leg muscle of WBOC. The relative mRNA expression of H-FABP was measured by real-time PCR at 30, 60, 90, and 120 d. The IMF content significantly increased with age in both muscles. The mRNA expression level of H-FABP significantly decreased with age in both muscles of the three types of chickens. Moreover, a significant negative correlation between H-FABP abundance and IMF content in the leg muscles of WBOC (p = 0.035) was observed. The mRNA expression of H-FABP negatively correlated with the IMF content in both breast and leg muscles of BOC sat slaughter time.

[Analysis of reason and strategy for the failure of posterior pedicle screw short-segment internal fixation on thoracolumbar fractures].

OBJECTIVE: To analyze the reason and strategy for failure of posterior pedicle screw short-segment internal fixation on thoracolumbar fractures. METHODS: From March 2008 to December 2010,the clinical data of 18 patients with thoracolumbar fracture failed in posterior pedicle screw short-segment internal fixation were retrospectively analyzed. There were 11 males and 7 females with an average age of 37.2 years (ranged, 19 to 63). The time from the first operation to complication occurrence was from 6 to 44 months with an average of 14.3 months. Of them,fusion failure was in 7 cases (combined with screw breakage in 4 cases), the progressive neuro-dysfunction was in 5 cases,the progressive lumbodorsal pain was in 6 cases. All 18 patients with kyphosis were treated with anterior internal fixation remaining posterior fixation (9 cases) and anterior internal fixation after posterior fixation removal (9 cases). RESULTS: All the patients were followed up from 18 to 50 months with an average of 30.5 months. No intetnal fixation loosening and breakage were found, moreover, X-ray and lamellar CT showed bone healing well. Preoperative, postoperative at 3 months and at final follow-up, ODI score was respectively 31.6+/-5.1, 8.6+/-5.7, 8.3+/-3.2; VAS score was respectively 7.2+/-2.3, 2.3+/-0.7, 2.1+/-1.1; kyphosis angle was respectively (-21.2/-+7.8 degreeso, (-5.3+/-6.8 degrees ), (-5.8+/-7.8 )degrees. Compared with preoperative data ,above-listed items had obviously ameliorated(P<0.05). CONCLUSION: Treatment of thoracolumbar fracture with posterior pedicle screw short-segment internal fixation may result in the complications such as bone nonunion ,internal fixation breakage and progressive kyphosis. Anterior reconstruction may be a good strategy for the failure of posterior operation.