Different intestinal tropism of the G2b Taiwan porcine epidemic diarrhea virus-Pintung 52 strain in conventional 7-day-old piglets.

The group 2b (G2b) porcine epidemic diarrhea virus (PEDV) that emerged in 2013 has since caused devastating diseases and economic loss. The full-length genome of the G2b Taiwan PEDV-Pintung 52 (PEDV-PT) strain and its intestinal tropism by evaluating the pathological changes in the original PEDV-PT infected field piglet and orally inoculation of either 10, 103, or 105 50% tissue culture infective dose/mL (TCID50/mL) of the plaque-purified PEDV-PT-Passage 5 (P5) in 7-day-old conventional piglets were analyzed. Phylogenetic analysis of the full-length genome indicated that the G2b Taiwan PEDV-PT strain was closely related to the North American G2b PEDV strains. Some pathological features of the G2b Taiwan PEDV-PT infection, including the absence of lesions and antigen signal in the crypt epithelial cells of the jejunum and ileum and in the villus enterocytes of the duodenum and colon, were different from those of infections by the North American G2b PEDV strains. This difference in the intestinal tropism of the G2b Taiwan PEDV-PT strain highlights the importance of studying the pathogenicities of different PEDV variants. Moreover, similar distributions of PEDV antigens and lesions in the G2b Taiwan PEDV-PT infected field piglet and its plaque-purified isolate, PEDV-PT-P5, inoculated piglets indicating that the plaque-purified PEDV-PT-P5 viral stock could facilitate the preclinical evaluation of vaccines and other interventions aimed at preventing the G2b PEDV infection.

The relationship of family characteristics and bipolar disorder using causal-pie models.

Many family characteristics were reported to increase the risk of bipolar disorder (BPD). The development of BPD may be mediated through different pathways, involving diverse risk factor profiles. We evaluated the associations of family characteristics to build influential causal-pie models to estimate their contributions on the risk of developing BPD at the population level. We recruited 329 clinically diagnosed BPD patients and 202 healthy controls to collect information in parental psychopathology, parent-child relationship, and conflict within family. Other than logistic regression models, we applied causal-pie models to identify pathways involved with different family factors for BPD. The risk of BPD was significantly increased with parental depression, neurosis, anxiety, paternal substance use problems, and poor relationship with parents. Having a depressed mother further predicted early onset of BPD. Additionally, a greater risk for BPD was observed with higher numbers of paternal/maternal psychopathologies. Three significant risk profiles were identified for BPD, including paternal substance use problems (73.0%), maternal depression (17.6%), and through poor relationship with parents and conflict within the family (6.3%). Our findings demonstrate that different aspects of family characteristics elicit negative impacts on bipolar illness, which can be utilized to target specific factors to design and employ efficient intervention programs.

A note on testing for intervention effects on binary responses.

OBJECTIVES: In some circumstances controlled trials are not feasible and treatments can only be evaluated using clinical databases. Here we consider the situation where treatment is introduced at a particular calendar time and can only be evaluated by comparison with historical controls. In these circumstances Heuer and Abel recommended using change-point methods to search for change in characteristics over the whole study period rather than simply comparing treated and untreated patients. Their recommendation is to only conclude that the intervention had an effect if a change-point could be demonstrated close in time to the introduction of the new treatment. This reduces the risk of false positives caused by confounding changes in population characteristics or changes in patient management. For binary data we develop a method that follows their philosophy and apply it to an observational study in the treatment of pin sites after orthopaedic surgery. METHODS: Tests for change in binomial probabilities based on Brownian bridge and Hansen's approximation for maximally selected chi(2) statistics are compared to an exact test by Worsley. The approximate method is generalized to logistic regression models allowing for covariates. RESULTS: The agreement of the exact and approximate method is good for sample sizes of 100 or more. The actual test size of the Hansen approximate test allowing for covariates is close to the nominal level, whereas the Brownian bridge approximation is slightly conservative. The change in pin site treatment significantly reduces the risk of infection for both adults and children. CONCLUSIONS: We consider the Hansen approximation to provide a very good and very simple method for obtaining the p-value when testing for a change in binary data event probabilities, with or without covariates.

Hepatitis C virus core protein recruits nucleolar phosphoprotein B23 and coactivator p300 to relieve the repression effect of transcriptional factor YY1 on B23 gene expression.

Hepatitis C virus (HCV) core has a pleiotropic effect on various promoters. In this study, we found that the expression of nucleolar phosphoprotein B23 was enhanced in HCV core-expressing cells and, moreover, HCV core interacts directly with the C-terminal end of B23. Using sucrose gradient centrifugation analysis and immunoprecipitation assays, HCV core was found in a large complex containing B23 and its interacting partner transcription factor YY1. Both B23 and HCV core associated with YY1 in the central GA/GK-rich and C-terminal zinc finger domain. These physical interactions between core, B23, and YY1 led to ternary complex formation that was bound to the YY1 response element. In a transient cotransfection experiment, relief of the trans-suppression activity of YY1 on the YY1-response element-driven reporter by core and B23 was found. This is also true when examining the effects of these three constructs on the B23 promoter-driven reporter. Additionally, chromatin immunoprecipitation assays indicated that a transcriptional activation complex consisting of core, together with B23, p300, and YY1, was recruited to the YY1 response element of B23 promoter, and this probably occurred through complex formation between core and these three cellular transcription regulators. This is different from the situation in the absence of core, where YY1 and histone deacetylase 1, but not B23 and p300, were associated on the YY1 element as the transcription repression complex. Together, our results indicate that HCV core can recruit B23 and p300 to relieve the repression effect of YY1 on B23 promoter activity, a property that requires the intrinsic histone acetyltransferase activity of p300. Thus, because these three core-associated cellular transcription regulators have a multitude of cellular interacting proteins and are involved in a versatility of cellular processes, the complex formation described here may partially account for the pleiotropic effects of core protein on gene expression and cellular function in HCV-infected cells.

Gene synthesis by a LCR-based approach: high-level production of leptin-L54 using synthetic gene in Escherichia coli.

Synthetic genes are very useful in genetic and protein engineering. Here we propose a general method for construction of synthetic genes. Short oligonucleotides are joined through ligase chain reaction (LCR) in high stringency conditions to make "unit fragments" which are then fused to form a full-length gene sequence by polymerase chain reaction. The procedure is simple and accurate and does not place constraints on sequence and length. In this report, a recombinant leptin gene was synthesized according to the codon preference of Escherichia coli. Besides, a substitution of the only Met at position 54 for Leu and an addition of a Met at the N-terminus were introduced in the synthetic gene. The gene was cloned in the pQE-31 expression vector and was expressed in E. coli. A large amount of recombinant leptin containing 6 x His tag was produced and purified by Ni-NTA affinity column. Finally, intact leptin-L54 was released after removing the tag by CNBr cleavage at the Met residue.

Performance of a neodymium yttrium aluminum borate green laser pumped with a fiber-coupled laser diode.

Using a space-dependent rate-equation model and considering the effect of pump beam quality, the optimum pump condition for a fiber-coupled diode end-pumped neodymium yttrium aluminum borate laser has been determined. Under optimum pump conditions, greater than 10% optical-to-optical conversion efficiency was obtained when the laser was pumped by a fiber-coupled laser diode with good beam quality. The influence of pump beam quality on the conversion efficiency is also discussed.

Single-Mode Operation and Frequency Doubling of Fiber-Coupled Diode Butt-Coupling-Pumped Nd:YVO(4) Lasers.

We present a simple way to achieve single-frequency operation by using a fiber-coupled diode butt-coupling-pumped Nd:YVO(4) laser in a flat-flat cavity. Single-mode outputs of 620 and 260 mW for fundamental and second-harmonic wavelengths were obtained when the laser was pumped by an 1100-mW fiber-coupled laser diode. Experimental results show that thermal effects provide not only a stable resonator with a good overlap of laser mode and pump size but also enhance single-frequency performance.

Physical mapping of RFLP markers on four chromosome arms in maize using terminal deficiencies.

Terminal deficiencies (TDs) generated by the r-X1 deletion system in maize were used to physically map RFLP markers on the short arm of chromosome 2 (2S) and the long arm of chromosome 6 (6L), chromosome 8 (8L), and chromosome 10 (10L). Five TDs on 2S, 8 on 6L, 10 on 8L, and 20 on 10L were isolated using the recessive morphological markers lg1, py1, j1(gl18), and sr2, respectively, for selection. Two exceptional TDs on 2S and 8L also have a second breakpoint on the long arm of chromosome 2 (2L) and 8L, respectively. The physical mapping of RFLP probes in relation to TD breakpoints was done by Southern hybridization. The five TDs on 2S divide chromosome 2 into four regions, all of which are distinguishable by RFLP markers. Likewise, three remaining chromosome arms are divided by TDs into RFLP-marked regions: 8 TDs divide 6L into five regions, 10 TDs divided 8L into seven regions, and 20 TDs divide 10L into three regions. The linear order of the physical map of 6L and 8L is consistent with that of the genetic maps, but that of 2L and 10L is not. Four groups of markers on 2S as well as 2L, and two on 10L are in reverse order in the physical map compared with the genetic maps. Other intriguing results are that breakpoints of TDs on 6L and 8L are distributed throughout the selected region, but most of those on 2L and 10L cluster in a region near the centromere: a single TD arose after fertilization.

DNA triplex formed by d-A-(G-A)7-G and d-mC-(T-mC)7-T in aqueous solution at neutral pH.

Evidence from UV spectroscopic melting experiments indicated that the DNA oligonucleotide 5'-d-A-(G-A)7-G-3' (1), a repeating AG sequence found in the human genome, and its complement 5'-d-mC-(T-mC)7-T-3' (mC: 5-methyl-C) (2), can form both a triplex (with a Tm = 44 degrees C) and a duplex (with a Tm = 69 degrees C) around physiological pH (7.2) in micromolar concentration solution with 0.1 M NaCl. In addition, the triplex can be detected at a pH as high as 8.4 (Tm = 27 degrees C). The stability of the triplex formed by 1 and 2, as monitored by UV melting experiments, is found to increase as the pH is lowered from 8.4 (Tm = 27 degrees C) to 6 (Tm = 79 degrees C). However, the stability of the duplex, formed by 1 and 2, is found to be unchanged (the Tm is approximately 69 degrees C) in the same pH range. There is no triplex, as observed by UV with the oligonucleotides having identical sequences, when the regular cytosine base was used [d-C-(T-C)7-T, (3)] at a pH > 6 in the same concentration range. The stoichiometric ratio of the triplex formed with 1 and 2 is also found to be 1:2 by means of a UV mixing titration study. This result suggests that the conformation of the triplex of 1 and 2 also involves T.A.T and mC+.G.mC base triads. The formation of a triplex by 1 and 2 can be observed by native gel electrophoresis in submicromolar conditions with magnesium ion present. The results of this study strongly support the theory that replacement of regular cytidine nucleotides by 5-methylcytosine nucleotides facilitates the formation of the DNA triplex at physiological pH. This could thus be used an an anti-gene probe via the formation of triplex under the conditions described.

Stability of ondansetron hydrochloride injection in extemporaneously prepared oral solutions.

The stability of ondansetron hydrochloride in extemporaneously prepared oral solutions containing orange juice, cola, or cherry syrup was determined. Solutions were prepared by adding ondansetron hydrochloride to orange juice, cola, or cherry syrup to produce ondansetron concentrations of 0.267 and 0.067 mg/mL in orange juice or cola and 0.533 mg/mL in cherry syrup. The ondansetron concentration in orange juice and cola solutions was assayed at the time of preparation and at 30 minutes and one hour. The cherry syrup solution was stored at both 3-5 and 25-27 degrees C, with the ondansetron concentration being determined at the time of preparation and daily for seven days. All the solutions were prepared in triplicate. Ondansetron concentrations were measured by stability-indicating high-performance liquid chromatography. At each time interval, the mean ondansetron concentration remained > or = 97% of the initial measurement for all solutions. The appearance and color of the solutions did not change. Ondansetron hydrochloride was stable for at least one hour in orange juice or cola and at least seven days in cherry syrup.

Stability of ondansetron in large-volume parenteral solutions.

OBJECTIVE: To determine the stability of ondansetron hydrochloride in large-volume parenteral solutions under four storage and time-period conditions. DESIGN/METHOD: Ondansetron was added to each of the following commercially available solutions to make final concentrations of approximately 24 and 96 micrograms/mL: NaCl 0.9%, D5W, and lactated Ringer's solution. SETTING: University analytical laboratory. MAIN OUTCOME MEASURES: Each solution was studied at both concentrations under the following conditions: (1) 1 day refrigerated, 2 days room temperature; (2) 7 days refrigerated, 2 days room temperature; (3) 14 days refrigerated, 2 days room temperature; and (4) 14 days room temperature. All solutions were exposed to fluorescent light when under room temperature conditions and were studied in triplicate. Ondansetron concentrations of samples were obtained periodically throughout each storage/time condition via a specific, stability-indicating HPLC method. RESULTS: A clinically significant loss of concentration was defined as greater than 10 percent decrease from the initial concentration. In all solutions and at both concentrations studied, the mean ondansetron concentration was greater than or equal to 90 percent under all storage and time conditions. CONCLUSIONS: Ondansetron can be stored and administered in these solutions without loss of potency.

The effects of short-term social support from dentists on the stress experienced by parents of pediatric dental patients.

Parents of pediatric dental patients were observed in a university clinic waiting room as they were greeted by student dentists. Soon after the dentist left with the child, the parents completed a series of scales assessing their perceived social support from the dentist, their anxiety and distress levels, and their confidence in the dentist. Regression analysis indicated that perceived social support from the dentist was associated negatively with parental distress regarding the dental treatment (beta = .-45, P < .01) and parental state anxiety (beta = .-37, P < .01). In addition, social support was associated positively with parent's confidence in the dentist beta = .54, P < .01). Possible mediational paths and clinical implications are discussed.

Some variables affecting dentists' assessment of patients' distress.

Dental students estimated the pain distress and discomfort of their patients after patients received a Class II restoration (simple filing). Dentist's estimates of patient pain accounted for 36% of the variance in patient self-report responses. Dentist accuracy was significantly lower, however, for estimates of anxiety and distress. Dentist accuracy was unaffected by patient's degree of self-monitoring. There was a tendency for female dentists to demonstrate greater accuracy than male dentists, but this effect was nonsignificant. In addition, dentists' accuracy in assessing overall patient discomfort was significantly lower in that segment of the treatment procedure that was most stressful for the dentists. This article discusses theoretical explanations for this latter effect.

Similarity of behavior within addict couples. Part II. Addiction-related variables.

The similarity of behavioral patterns within addict couples before, during, and after a relationship is the focus of this paper. The correlations between partners in real and artificially constructed (pseudo) couples for employment, criminal activities, and other behaviors are examined. The effect of a relationship on behavior is also assessed before, during, and after treatment. Significant correlations during real relationships were found for employment, welfare, and illegal income. Joint couple entry into treatment appears to be more effective for women than men. Similarity within couples disappears when the relationship ends.

Similarity of behavior within addict couples. Part I. Methodology and narcotics patterns.

The development and initial assessment of a methodology for measuring the similarity of behavioral patterns within addicted couples over the duration of a relationship is presented. Two subsamples of addicts from methadone maintenance programs in southern California were established: one of couples who had a real relationship during their addiction and treatment careers, and a group of pseudocouples (matched as closely as possible) created from unrelated clients in the same programs. Narcotics use, abstinence, and support patterns are analyzed for couples before, during, and after their relationship. When it occurs during the relationship, treatment effects are also analyzed.

The efficient assessment of need for cognition.

A short form for assessing individual differences in need for cognition is described.