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Objective To investigate the potential of EEG multiscale entropy and complexity as biomarkers in infantile spasms. Methods We collected EEG data retrospectively from 16 newly diagnosed patients, 16 age- and gender-matched healthy controls, and 15 drug-resistant patients. The multiscale entropy (MSE) and total EEG complexity before anti-epileptic drug (AED) treatment, before adrenocorticotropic hormone (ACTH) treatment, 14 days after ACTH therapy, and after 6 months of follow-up were calculated. Results The total EEG complexity of 16 newly diagnosed infantile spasms patients was lower than the 16 healthy controls (median [IQR]: 351.5 [323.1–388.1] vs 461.6 [407.7–583.4]). The total EEG complexity before treatment was higher in the six patients with good response to AED than the 10 patients without response (median [IQR]: 410.0 [388.1–475.0] vs 344.5 [319.6–352.0]). The total EEG complexity before and after 14-days of ACTH therapy was not different between 13 ACTH therapy responders and nine non-responders. After 6-months follow-up, the total EEG complexity of ACTH therapy responders were higher than non-responders (median [IQR]: 598.5 [517.4–623.3] vs 448.6 [347.1–536.3]). Conclusions The total EEG complexity before AED and 6 months after ACTH are associated with spasm-freedom. Significance The total EEG complexity is a potential biomarker to predict and monitor the treatment effect in infantile spasms.

Objective Human papillomavirus (HPV) testing as the primary cervical cancer screening followed by reflex cytology if high-risk HPV is present (hrHPV+) is recently adopted in some countries. However, reflex cytology's sensitivity is variable, and a suitable triage approach for hrHPV+ remains controversial. Here, we compared the performance of three triage tools in hrHPV+ women. Methods Three triage tools—cytology, HPV16/18 genotyping, and DNA methylation biomarker PAX1m—were analyzed for their clinical performance in hrHPV+ women. In addition, women without cervical cancer at enrollment were followed for histologically confirmed high-grade cervical intraepithelial neoplasia or worse (CIN3+) annually using Papanicolaou smear. Results Of 4762 women aged ≥20 years enrolled, 502 (10.5%) were hrHPV+. PAX1m and cytology demonstrated similar accuracy (>90%), sensitivity (>78%), and specificity (>92%) as triage tools in 429 hrHPV+ women aged 30–64 years. PAX1m had better accuracy and specificity (91.6% and 92.5%, respectively) than HPV16/18 (76.9% and 76.8%, respectively). The incidence of CIN3+ among hrHPV+ women was 10.7 cases/1000 person-years. The incidence was significantly greater in PAX1m-positive women than in PAX1m-negative women. Conclusions PAX1m has comparable clinical performance to cytology and better accuracy and specificity than HPV16/18 as the triage tool for detecting CIN3+ in hrHPV+ women. The PAX1m assay is thus a promising molecular-based triage tool for early detection of CIN and predicting disease progression in hrHPV+ women. It can be especially useful in countries where adequate cytology-based infrastructure is lacking, such as some Southeast Asian countries, for cervical cancer screening and prevention.

Objectives This study investigated whether the methylation of ZNF582, PAX1, SOX1, NKX6.1, and PTPRR genes in oral scrapings could be used to detect oral dysplasia and oral cancer and to predict oral cancer recurrence. Materials and methods Oral scrapings were collected from 65 normal oral mucosa subjects, 107 oral precancer patients, and 95 oral squamous cell carcinoma patients. Methylation levels of the five genes were quantified by real-time methylation-specific PCR after bisulfite conversion. Results Among the five tested genes, methylated ZNF582 (ZNF582m) and PAX1 (PAX1m) were found to be appropriate biomarkers for oral dysplasia and oral cancers. ZNF582m could detect mild dysplasia or worse oral lesions with the sensitivity and specificity being 0.85 and 0.87, respectively. PAX1m performed better in identifying moderate dysplasia or worse oral lesions with the sensitivity and specificity being 0.72 and 0.86, respectively. Moreover, the methylation levels and positive rates for ZNF582m and PAX1m were increased when disease severity increased. Thus, they may be applicable as a triage tool for patients with abnormal visual oral examinations. After cancer excision, both ZNF582m and PAX1m levels decreased. However, their levels increased again at the subsequently recurrent sites in some patients approximately 3–4 months before cancer recurrence. Finally, areca-quid chewing alone and in combination with cigarette smoking or alcohol drinking were found to be correlated with ZNF582 and PAX1 hypermethylation. Conclusion We conclude that hypermethylated ZNF582 and PAX1 are effective biomarkers for the detection of oral dysplasia and oral cancer and for the prediction of oral cancer recurrence.

This study assessed whether hypermethylated ZNF582 and PAX1 genes in oral scrapings are correlated with the progression and prognosis of oral squamous cell carcinoma (OSCC).Methylation levels of ZNF582 and PAX1 genes in oral scrapings, collected from the cancer and adjacent normal oral mucosal sites of 80 OSCC patients before surgical cancer excision, were quantified using real-time methylation-specific PCR after bisulfite conversion.Both the mean methylation (M)-indices of ZNF582 and PAX1 genes in oral scrapings were significantly higher at the cancer sites than at the adjacent normal oral mucosal sites (both P .001). In the oral scrapings collected from the adjacent normal oral mucosal sites, the higher M-index of methylated ZNF582 (ZNF582Hypermethylated ZNF582 and PAX1 genes in the oral scrapings collected from adjacent normal oral mucosal sites rather than cancer sites are associated with aggressive progression and poor prognosis of OSCC.

Electroencephalography (EEG) is frequently used in functional neurological assessment of children with neurological and neuropsychiatric disorders. Multiscale entropy (MSE) can reveal complexity in both short and long time scales and is more feasible in the analysis of EEG. Entropy-based estimation of EEG complexity is a powerful tool in investigating the underlying disturbances of neural networks of the brain. Most neurological and neuropsychiatric disorders in childhood affect the early stage of brain development. The analysis of EEG complexity may show the influences of different neurological and neuropsychiatric disorders on different regions of the brain during development. This article aims to give a brief summary of current concepts of MSE analysis in pediatric neurological and neuropsychiatric disorders. Studies utilizing MSE or its modifications for investigating neurological and neuropsychiatric disorders in children were reviewed. Abnormal EEG complexity was shown in a variety of childhood neurological and neuropsychiatric diseases, including autism, attention deficit/hyperactivity disorder, Tourette syndrome, and epilepsy in infancy and childhood. MSE has been shown to be a powerful method for analyzing the non-linear anomaly of EEG in childhood neurological diseases. Further studies are needed to show its clinical implications on diagnosis, treatment, and outcome prediction.

Background Effective biomarkers for oral cancer screening are important for early diagnosis and treatment of oral cancer. Methods Oral epithelial cell samples collected by mouth rinse were obtained from 65 normal control subjects, 108 patients with oral potentially malignant disorders, and 94 patients with oral squamous cell carcinoma (OSCC). Methylation levels of zinc-finger protein 582 (ZNF582) and paired-box 1 (PAX1) genes were quantified by real-time methylation-specific polymerase chain reaction after bisulfite conversion. Results An abrupt increase in methylated ZNF582 (ZNF582m) and PAX1 (PAX1m) levels and positive rates from mild dysplasia to moderate/severe dysplasia, indicating that both ZNF582m and PAX1m are effective biomarkers for differentiating moderate dysplasia or worse (MODY+) oral lesions. When ZNF582m/PAX1m tests were used for identifying MODY+ oral lesions, the sensitivity, specificity, and odds ratio (OR) were 0.65/0.64, 0.75/0.82, and 5.6/8.0, respectively. Conclusion Hypermethylated ZNF582 and PAX1 genes in oral epithelial cells collected by mouth rinse are effective biomarkers for the detection of oral dysplasia and oral cancer.

Objective Tourette syndrome is a developmental neuropsychiatric disorder in children, and abnormal corticobasal ganglion connectivity is implied for the pathophysiology. Multiscale entropy, an entropy-based method to measure dynamic complexity at multiple temporal scales, is helpful to disclose the information of brain connectivity. This preliminary study investigated the complexity of resting-state electroencephalogram signals using multiscale entropy in children with Tourette syndrome. Method Resting-state electroencephalographic (EEG) signals were analyzed by sample entropy and multiscale entropy methods in 10 children with Tourette syndrome and 10 healthy gender- and age-matched controls. Results Except for the Fp2 channel, the complexity index values in all channels were reduced in children with Tourette syndrome compared with those in normal controls. A statistically significant reduction in EEG complexity was found in the bilateral central, parietal, occipital, and left temporal regions, indicating disturbed brain connectivity in Tourette syndrome. Although there was no difference of complexity in the higher frequency spectra, there was a statistically significant difference of complexity in lower frequency in F3 channel, pointing to the importance of examining a range of time scales in exploring EEG signals. Conclusions Our preliminary study demonstrated that EEG complexity was significantly lower in children with Tourette syndrome than in normal controls. This difference may serve as a marker of disturbed brain connectivity in such individuals and suggests that further clinical studies are warranted. (PsycINFO Database Record

ObjectivesDNA methylation is a potential biomarker for early cancer detection. Previous studies suggested that the methylations of several genes are promising markers for the detection of cervical intraepithelial neoplasia at grade III or worse (CIN3+). The purpose of the present study was to explore the feasibility of these DNA methylation testing in cervical cancer screening.MethodsA total of 443 women were recruited from the Yuan’s General Hospital. Cervical scrapings were collected for Papanicolaou (Pap) test by using cervical brushes, and the cytological data were used for analysis. The residual cells on the brush were preserved in phosphate-buffered saline solution at 4°C until DNA extraction. Then, the extracted DNA were used for molecular tests, which included human papillomavirus typing and quantification of the methylation levels for PAX1, SOX1, and NKX6-1 genes. Subjects who had abnormal Pap test results underwent colposcopy or biopsy with subsequent conization or major surgery when biopsy results revealed CIN2+. The final diagnosis for this group was confirmed by colposcopy or pathological examination. The study was approved by the institutional review board of Yuan’s General Hospital, and all the molecular tests were performed by ISO17025 certified laboratories.ResultsThe sensitivity of PAX1m and SOX1m was greater than 80%, and the specificity of PAX1m and NXK6-1m was greater than 80% for the detection of CIN3+ lesions. PAX1m detection alone had a sensitivity and specificity of 86% and 85%, respectively, whereas when used as a cotest with the Pap test, the sensitivity and specificity were 89% and 83%, respectively.ConclusionsPAX1m showed great potential as a biomarker for cervical cancer screening. When incorporating PAX1m detection into current screening protocol, the efficacy of screening could be greatly improved. Moreover, unnecessary referral for colposcopy and biopsy could be reduced up to 60%. However, prospective population-based studies are necessary for further implementation of this screening program.

The characteristics of hybrid gadolinium oxide nanocrystal (Gd 2 O 3 -NC) and gadolinium oxide charge trapping (Gd 2 O 3 -CT) memories were investigated with different Gd 2 O 3 film thickness. By performing the rapid thermal annealing on Gd 2 O 3 films with different thickness, the Gd 2 O 3 -NCs with the diameter of 6–9 nm for charge storage, surrounded by the amorphous Gd 2 O 3 ( α -Gd 2 O 3 ) layer, were formed. The α -Gd 2 O 3 layer was considered to be the charge trapping layer, resulting in the large memory window of Gd 2 O 3 -NC/CT memories with thick Gd 2 O 3 film. The charge trapping energy level of the Gd 2 O 3 -NCs and α -Gd 2 O 3 layer was extracted to be 0.16 and 0.45 eV respectively by using the temperature-dependent retention measurement. Further, after a 10 6 program/erase cycling operation, the memory with thin Gd 2 O 3 film can be predicted to sustain a 94% memory window of the first cycling one while the memory with thick Gd 2 O 3 film suffered from a 30% charge loss because of the traps within the α -Gd 2 O 3 layer. The Gd 2 O 3 film thickness of 10 nm was optimized to exhibit superior performances of the Gd 2 O 3 -NC/CT memory, which can be applied into the nonvolatile memory.

Population and test characteristics by histologic category in training set. (PPTX 47 kb)

Background Carbamazepine, an anticonvulsant and a mood-stabilizing drug, is the main cause of the Stevens–Johnson syndrome (SJS) and its related disease, toxic epidermal necrolysis (TEN), in Southeast Asian countries. Carbamazepine-induced SJS–TEN is strongly associated with the HLA-B*1502 allele. We sought to prevent carbamazepine-induced SJS–TEN by using HLA-B*1502 screening to prospectively identify subjects at genetic risk for the condition. Methods From 23 hospitals in Taiwan, we recruited 4877 candidate subjects who had not taken carbamazepine. We genotyped DNA purified from the subjects' peripheral blood to determine whether they carried the HLA-B*1502 allele. Those testing positive for HLA-B*1502 (7.7% of the total) were advised not to take carbamazepine and were given an alternative medication or advised to continue taking their prestudy medication; those testing negative (92.3%) were advised to take carbamazepine. We interviewed the subjects by telephone once a week for 2 months to monitor them ...

DNA methylation is a potential biomarker for early cancer detection. Previous studies suggested that the methylations of several genes are promising markers for the detection of cervical intraepithelial neoplasia at grade III or worse (CIN3+). The purpose of the present study was to explore the feasibility of these DNA methylation testing in cervical cancer screening.A total of 443 women were recruited from the Yuan's General Hospital. Cervical scrapings were collected for Papanicolaou (Pap) test by using cervical brushes, and the cytological data were used for analysis. The residual cells on the brush were preserved in phosphate-buffered saline solution at 4°C until DNA extraction. Then, the extracted DNA were used for molecular tests, which included human papillomavirus typing and quantification of the methylation levels for PAX1, SOX1, and NKX6-1 genes. Subjects who had abnormal Pap test results underwent colposcopy or biopsy with subsequent conization or major surgery when biopsy results revealed CIN2+. The final diagnosis for this group was confirmed by colposcopy or pathological examination. The study was approved by the institutional review board of Yuan's General Hospital, and all the molecular tests were performed by ISO17025 certified laboratories.The sensitivity of PAX1 and SOX1 was greater than 80%, and the specificity of PAX1 and NXK6-1 was greater than 80% for the detection of CIN3+ lesions. PAX1 detection alone had a sensitivity and specificity of 86% and 85%, respectively, whereas when used as a cotest with the Pap test, the sensitivity and specificity were 89% and 83%, respectively.PAX1 showed great potential as a biomarker for cervical cancer screening. When incorporating PAX1 detection into current screening protocol, the efficacy of screening could be greatly improved. Moreover, unnecessary referral for colposcopy and biopsy could be reduced up to 60%. However, prospective population-based studies are necessary for further implementation of this screening program.

This work proposes synthesis methods to design the mechanism that is adjustable for tracing variable circular arcs with prescribed velocities. The constraint equations and useful properties of the desired mechanism are derived by using the concept of cross-ratio. The desired mechanism can be a crank-rocker or a slider-crank mechanism. The coupler curve of the mechanism, if necessary, can be symmetrical to itself. The practical applications of the mechanism are also discussed.

Overproduction of the short-chain dehydrogenase/reductase PTR1 confers resistance to the dihydrofolate reductase inhibitor methotrexate in the protozoan parasite Leishmania. Genetic analysis has previously implicated PTR1 in pterin and folate metabolism. PTR1 was purified from a fusion protein expressed in Escherichia coli. Purified PTR1 exhibits NADPH-dependent biopterin, dihydrobiopterin, folate, and dihydrofolate reductase activities. The highest activity was found with the most oxidized pterins. The active protein was found to be a tetramer as demonstrated by gel-filtration chromatography. Kinetic constants (K(m)), as determined by double-reciprocal plots, were calculated for NADPH and for several of PTR1's substrates. The PTR1 of Leishmania tarentolae had a K(m) of 16.9 microM for the cofactor NADPH and K(m) values ranging from 3.5 to 85 microM for the various substrates. The dissociation constant (KD), as determined by fluorescence titration, for NADPH was estimated to be 130 microM. The biochemical characterization of this important and novel enzyme involved in folate and pterin metabolism of Leishmania should be useful for structure-function analysis and for developing specific inhibitors against this putative important chemotherapeutic target.

Design equations for spherical four-bar linkages to trace a coupler curve with two prescribed cusps are derived in this study by using a special case of spherical Burmester curves. For the case in which the tangents at two cusps are also prescribed, an analytical method is proposed with the aid of the foregoing design equations associated with the concept of the spherical cross ratio and spherical Bobillier theorem. The proposed method is straightforward and quite useful for those cases in which the tangents at two cusps are neither on the same plane nor on parallel planes. Numerical examples are also provided for illustrating the entire technique.

The gadolinium oxide nanocrystal (Gd2O3-NCs) memory has been demonstrated with large memory window and good reliabilities. It has been proven that the charges are stored in the crystallized Gd2O3-NCs which are surrounded by trapping-free amorphous phase of Gd2O3 film. The calculated deep trapping energy level is about 1.7eV below the conduction of the Gd2O3-NCs, and the energy band diagram of the Gd2O3-NCs memory has been proposed. In this study, we optimize the thickness of Gd2O3 thin film to investigate the effect on the charge storage mechanism. The thickness of Gd2O3 thin film and blocking oxide could simultaneously dominate the operation speed and charge loss phenomenon. Thus, the most suitable film thicknesses for the Gd2O3-NCs memory were proposed for future mass production nonvolatile memory application.

A new method is proposed to derive the special cases that the center- and circle-point curves of PP-PP multiply separated positions degenerate into the combinations of points, lines and circles. Analytical equations for the construction of the Burmester curves are presented in a simpler form with four parameters. All conditions that yield degenerate forms of the equations are concluded by investigating the parameters. Degenerate Burmester curves are then thoroughly derived from various combinations of the conditions. The results of this work are more complete than existing papers, and presented concisely in tables. Applications of the degenerate curves are also illustrated by an example.

With a pole and two instant centres specified on a unit sphere, a geometrical construction is proposed to determine the spherical centre- and circle-points. The explicit equations of the corresponding spherical centre- and circle-point curves of the PP-PP case are thus obtained by using spherical trigonometry. All break-ups of these spherical centre- and circle-point curves are directly generated from the explicit equations. Through an exhaustive search, it was found that there are four kinds of degenerated curves, i.e. (1) two points and a great circle, (2) two spherical ellipses and a great circle, (3) three orthogonal great circles and (4) a sphere. Two design examples are presented.

In this paper a cross ratio theory applied in sphere geometry and its application on the spherical mechanism is presented. By expanding the cross ratio concept in plane geometry, cross ratio is proved to be equally valid in sphere geometry. Then, an alternative form of spherical cross ratio is proposed. An application of spherical cross ratio for the design of spherical four-bar mechanisms is presented.

The analytical methods employed for synthesizing the Watt-I mechanisms, which generate closed coupler curves with up to four cusps, are presented in this paper. The necessary conditions for a Watt-I mechanisms to generate a cusp are classified. Closed-form aolutions for synthesizing the mechanisms which generate up to three cusps and pass through up to three coupler positions are also proposed. The mechanism generating closed coupler curve with four cusps is synthesized by using an optimization technique.

In this study, new geometrical constructions and simplified equations are proposed to determine the center- and circle-point curves of four-bar linkages with specified first derivative of motion. Properties and degenerations of the curves can easily be investigated by the proposed methods. By using the degenerate curves, the equations for evaluating the Burmester centers are greatly simplified to cubic and quadratic equations. The methods proposed herein can be applied directly to the synthesis of linkages for motion generator, path generator, and function generator with specified first derivative of motion. Interesting applications are presented for illustrating the entire techniques.

It has been found that the path of every point on a moving plane has a cusp in the T-position of the second kind, which is in nature an instantaneous dwell-position. In this paper, cases which violate such a property are found, hence the motion of a moving plane in the vicinity of the instantaneous dwell-position is investigated. Interesting properties and their interrelationships are obtained, such as the number of consecutive contact points between the fixed polode and the moving polode, the paths of points on the moving plane, path tangent at the cusp, etc. The instant center of a link in the instantaneous dwell-position, in general, cannot be determined by the existing methods. However, based on the current properties, the instant center can be determined analytically. A geometric procedure presented here is able to determine the instant center of the stationary link of six-link mechanisms in the limit position.

Aims: Warfarin, a widely prescribed oral anticoagulant, is used for the prevention of thromboembolism. Several polymorphisms in VKORC1 have been shown to be associated with warfarin dose requirements. The frequencies of these VKORC1 polymorphisms display population differences; however, this has not been examined in many populations. In this study, we examined VKORC1 polymorphisms in five East-Asian populations (Han Chinese, Indonesian, Filipino, Thai and Vietnamese) and Indians. Materials & methods: A total of six SNPs in the VKORC1 gene (-1639G>A, 497T>G, 1173C>T, 1542T>G, 2255C>T and 3730G>A) were genotyped. Frequencies, linkage disequilibrium and haplotype structures of these six VKORC1 SNPs were analyzed. Results: Our data showed that 497T>G is only polymorphic in the Indian population. The 497G allele is very rare in the East-Asian populations (frequency < 1%). The remaining SNPs demonstrated high linkage disequilibrium and had similar frequencies and haplotype structures in all but the Indian population. The Indian population is mostly made up of the H7 haplotype (76%) while the rest of the recruited populations consisted of the H1 haplotype (>80%).

Absence epilepsy is an important epileptic syndrome in children. Multiscale entropy (MSE), an entropy-based method to measure dynamic complexity at multiple temporal scales, is helpful to disclose the information of brain connectivity. This study investigated the complexity of electroencephalogram (EEG) signals using MSE in children with absence epilepsy. In this research, EEG signals from 19 channels of the entire brain in 21 children aged 5-12 years with absence epilepsy were analyzed. The EEG signals of pre-ictal (before seizure) and ictal states (during seizure) were analyzed by sample entropy (SamEn) and MSE methods. Variations of complexity index (CI), which was calculated from MSE, from the pre-ictal to the ictal states were also analyzed. The entropy values in the pre-ictal state were significantly higher than those in the ictal state. The MSE revealed more differences in analysis compared to the SamEn. The occurrence of absence seizures decreased the CI in all channels. Changes in CI were also significantly greater in the frontal and central parts of the brain, indicating fronto-central cortical involvement of "cortico-thalamo-cortical network" in the occurrence of generalized spike and wave discharges during absence seizures. Moreover, higher sampling frequency was more sensitive in detecting functional changes in the ictal state. There was significantly higher correlation in ictal states in the same patient in different seizures but there were great differences in CI among different patients, indicating that CI changes were consistent in different absence seizures in the same patient but not from patient to patient. This implies that the brain stays in a homogeneous activation state during the absence seizures. In conclusion, MSE analysis is better than SamEn analysis to analyze complexity of EEG, and CI can be used to investigate the functional brain changes during absence seizures.

An analytical method for the synthesis of planar four-bar path generator with specified inclinations of path tangents at two cusps has been presented here. Simplified equations based on the concept of kinematic inversion and cross-ratio have been derived for the design problem. The desired four-bar linkages have been synthesized directly by using these analytical equations.

PTR1, the gene promoting MTX resistance following gene amplification or DNA transfection in Leishmania tarentolae and selected mutants, has been cloned and heavily overexpressed (>100 mg/liter) in Escherichia coli strain BL21 (DE3). Protein has been purified, essentially to homogeneity, in two steps, via ammonium sulfate precipitation and chromatography on DEAE-Trisacryl. The active proteins are tetramers and display optimal pteridine reductase activity at pH 6.0 using biopterin as substrate and NADPH as the reduced dinucleotide cofactor. 2,4-Diaminopteridine substrate analogues are strong competitive inhibitors (K(i) approximately 38 --> 3 nM) against the pterin substrate and both NADP(+) and folate are inhibitors although somewhat weaker. Dihydropteridines are poor substrates compared to the fully oxidized pteridine. Kinetic analysis affords the usual Michaelis constants and in addition shows that inhibition by NADP(+) allows the formation of ternary nonproductive complexes with folate. The kinetic results are consistent with a sequential ordered bi-bi kinetic mechanism in which first NADPH and then pteridine bind to the free enzyme. Sequence comparisons suggest that PTR1 belongs to the short-chain dehydrogenase/reductase (SDR) family containing an amino-terminal glycine-rich dinucleotide binding site plus a catalytic Y(Xaa)(3)K motif. In accord with this observation, the mutants K16A, Y37D, and R39A and the double mutants K17A:R39A and Y37D:R39A all show a two- to threefold lower binding affinity for NADPH and exhibit low or zero activity. Two Y(Xaa)(3)K regions are present in wild-type PTR1 at 152 and 194. Only Y194F gives protein with zero activity. This observation coupled with affinity labeling of PTR1 by oNADP(+) (2', 3'-dialdehyde derivative of NADP(+)) followed by NaBH(4) reduction, V8 protease digestion, and mass spectral analysis suggests that the motif participating in catalysis is that at 194. The mutation K198Q eliminates inactivation by oNADP(+) supporting the hypothesis that K198 is associated with nucleotide orientation, as has been demonstrated for similar lysine residues in other members of the SDR family.

Naturally occurring pteridines in eukaryotic systems usually contain 2-amino, 4-hydroxy 6-alkyl substituents composed of either a methylene-p-aminobenzoylglutamate (or polutamate) or a dihydroxypropyl group. The former class are known collectively as the folates and occur widely as reduced and 5-alkylated derivatives, in which form they participate in important metabolic one-carbon transfers (Blakley, 1984). The latter, known as biopterin,, also occurs in reduced forms and is an important cofactor in aromatic amino acid hydroxylations en route to the catecholamines (Shiman, 1985; Kaufman and Kaufman, 1985; n and Lovenberg, 1985) and in the nitrite synthase pathway (Marletta,, 1993). Dihydrofolate reductase (DHFR), dihydropteridine reductase (DHPR) and pteridine reductase (PTR1), best characterised from Leishmania, are three enzymes that initiate the reduction of a pteridine in association with a reduced dinucleotide cofactor. Their comparative reaction pathways are illustrated in Figure 1.

Mutations at the cytokinin biosynthesis locus (tmr) of Agrobacterium tumefaciens usually result in strains that induce tumors exhibiting the rooty phenotype associated with high auxin-to-cytokinin ratios. However, tobacco (Nicotiana tabacum cv Havana 425) leaf disc explants responded to tmr- mutant strain A356 by producing rapidly growing, unorganized tumors, indicating that these lines can grow in a cytokinin-independent fashion despite the absence of a functional tmr gene. Several methods have been used to characterize the physiological and cellular basis of this phenotype. The results indicate that tmr- tumors have a physiologically distinct mechanism for cytokinin-independent growth in comparison to tumors induced by wild-type bacteria. The cytokinin-independent phenotype of the tmr- transformants appears to be cell autonomous in nature: only the transformed cells and their progeny were capable of cytokinin-independent growth. Specifically, the tmr- tumors did not accumulate cytokinin, and clonal analysis indicated the tmr- transformed cells were not capable of stimulating the growth of neighboring nontransformed cells. Finally, the cytokinin-independent phenotype of the tmr- transformants was shown to be cold sensitive, whereas the wild-type tumors exhibited a cold-resistant cytokinin-independent phenotype. Potential mechanisms for this novel form of cytokinin-independent growth, including the role of the dehydrodiconiferyl alcohol glucosides found in both tumor types, are discussed.

Several complex phenotypic changes are induced when the transcription factor AmMYB308 is overexpressed in transgenic tobacco plants. We have previously shown that the primary effect of this transcription factor is to inhibit phenolic acid metabolism. In the plants that we produced, two morphological features were prominent: abnormal leaf palisade development and induction of premature cell death in mature leaves. Evidence from the analysis of these transgenic plants suggests that both changes resulted from the lack of phenolic intermediates. These results emphasize the importance of phenolic secondary metabolites in the normal growth and development of tobacco. We suggest that phenolic acid derivatives are important signaling molecules in the final stages of leaf palisade formation and that phenolic acid derivatives also play a prominent role in tissue senescence.

Seven kinds of flavones, β-sitosterol, β-sitosterol-D-glucoside, hesperidin, limonin, ferulic acid and succinic acid were isolated from the fruit peel of Citrus tankan Hayata.

Key Word IDX batwood; flavonoids; (2R,3R)-3-acctoxy-5,7dihydroxy-6- mcthylflavanoae; (2R,3R)-3,5.7-trihydroxy-6-methyltlavanone. Abstract-Two dihydroflavonols. (2R,3R)-3-aoetoxy-5,7dihydroxy-6-methyhlavanonc and (2R,3Rb3,5,7-trihydroxy- 6-mcthyllIavanone, accompanied by 12 known tlavottoids were isolated from the acetone extract of the heartwood of Pinus morri.wnicola. The major flavonoids include chrysin, pinobattksin and tectochrysin.

Background Opportunistic screening in hospitals is widely used to effectively reduce the incidence rate of cervical cancer in China and other developing countries. This study aimed to identify clinical risk factor algorithms that combine gynecologic examination and molecular testing (paired box gene 1 (PAX1) or zinc finger protein 582 (ZNF582) methylation or HPV16/18) results to improve diagnostic accuracy. Methods The delta Cp of methylated PAX1 and ZNF582 was obtained via quantitative methylation-specific PCR in a training set (57 CIN2− and 43 cervical intraepithelial neoplasia ≥grade 3 (CIN3+) women), and the individual and combination gene sensitivities and specificities were determined. The detection accuracy of three algorithms combining gynecologic findings and genetic test results was then compared in a randomized case-control study comprising 449 women referred for colposcopic examination by gynecologists in the outpatient department of Xiangya Hospital between November 2011 and March 2013. Results Significant association was observed between CIN3+ and methylated PAX1 or ZNF582 in combination with HPV16/18 (OR:15.52, 95 % CI:7.73–31.18). The sensitivities and specificities of methylated PAX1 or ZNF582 combined with HPV16/18 for CIN3+ women were 89.2 and 76.0 %, or 85.4 and 80.1 %, respectively. Of the three algorithms applied to cohort data and validated in the study, two indicated 100 % sensitivity in detecting cervical cancer and a low rate of referrals for colposcopy. Conclusions These algorithms might contribute to precise and objective cervical cancer diagnostics in the outpatient departments of hospitals in countries with high mortality and low screening rates or areas with uneven resource distribution. Electronic supplementary material The online version of this article (doi:10.1186/s13148-016-0232-3) contains supplementary material, which is available to authorized users.

The interpretation of equivocal Papanicolaou (Pap) smear results remains challenging, even with the addition of the high-risk human papillomavirus test (HPV-HR). Recently, methylated zinc finger protein 582 (ZNF582) (ZNF582 m ) was reported to be highly associated with cervical cancer. In this study, we compared the performance of ZNF582 m detection and HPV-HR genotyping in the triage of cervical atypical squamous cell of undetermined significance (ASC-US) and atypical squamous cell - cannot exclude a high-grade lesion (ASC-H). Two hundred and forty-two subjects with equivocal papanicolaou smear (Pap smear) results were recruited in this hospital-based and case-controlled study. The residual cervical cells in liquid-based cytological test (LBC) containers were used for genomic DNA extraction and then for ZNF582 m and HPV-HR detection. The level of ZNF582 m was quantified by real-time methylation-specific PCR after bisulfite conversion. The HPV-HR test was performed by using a nested multiplex PCR (NMPCR) assay that combines degenerate E6/E7 consensus primers and HPV type-specific primers. Significant associations were observed between ZNF582 m and the risk of cervical intraepithelial neoplasia grade 3 or higher (CIN3+; odds ratio = 15.52, 95% confidence interval (CI): 7.73 to 31.18). The sensitivity and specificity of ZNF582 m for women with CIN3+ were 82.43% and 76.79%, respectively. High sensitivity (99.33%) but low specificity (38.76%) was observed for HPV-HR. When combining both positive results of ZNF582 m and HPV-HR, the sensitivity and specificity were 82.43% and 81.55%, respectively. The sensitivity and specificity of ZNF582 m or HPV-16/18 were 89.19% and 70.24%, respectively. However, the sensitivity and specificity of ZNF582 m combined with HPV-16/18 (both ZNF582 m and HPV-16/18 positive results) were 59.46% and 94.64%, respectively. ZNF582 m provides a promising triage tool for women with ASC. To effectively manage ASC patients, a new strategy co-testing for ZNF582 m and HPV-16/18 genotyping was proposed. This strategy could reduce the number of patients referred for colposcopic examination and thus provide a feasible follow-up solution in the regions where colposcopy is not readily available. This strategy could also prevent women from experiencing unnecessary anxiety caused by HPV-HR.