RET mutational spectrum in Hirschsprung disease: evaluation of 601 Chinese patients

Rare (RVs) and common variants of the RET gene contribute to Hirschsprung disease (HSCR; congenital aganglionosis). While RET common variants are strongly associated with the commonest manifestation of the disease (males; short-segment aganglionosis; sporadic), rare coding sequence (CDS) variants are more frequently found in the lesser common and more severe forms of the disease (females; long/total colonic aganglionosis; familial).Here we present the screening for RVs in the RET CDS and intron/exon boundaries of 601 Chinese HSCR patients, the largest number of patients ever reported. We identified 61 different heterozygous RVs (50 novel) distributed among 100 patients (16.64%). Those include 14 silent, 29 missense, 5 nonsense, 4 frame-shifts, and one in-frame amino-acid deletion in the CDS, two splice-site deletions, 4 nucleotide substitutions and a 22-bp deletion in the intron/exon boundaries and 1 single-nucleotide substitution in the 5' untranslated region. Exonic variants were mainly clustered in RET the extracellular domain. RET RVs were more frequent among patients with the most severe phenotype (24% vs. 15% in short-HSCR). Phasing RVs with the RET HSCR-associated haplotype suggests that RVs do not underlie the undisputable association of RET common variants with HSCR. None of the variants were found in 250 Chinese controls.     

Two new flavones from Eupatorium coelestinum

Besides coumarin, nobiletin, lucidin dimethyl ether and 5,6,7,3′,4′-pentamethoxyflavone, two new highly oxygenated flavones were isolated from Eupatorium coelestinum. Their structures were determined by spectroscopic methods and alkaline degradations as 5,6,7,8,3′,4′,5′ -heptamethoxyflavone and 5,6,7,8,5′-pentamethoxy-3′,4′-methylenedioxyflavone.     

Specific Treatment Technologies for Removing Arsenic from Water

Arsenic (As) is a highly toxic metalloid found in ground and surface water. Arsenic contamination in drinking water leads to harmful effects on human health. To eliminate arsenic from drinking water, several technologies such as coagulation, adsorption, ion exchange, filtration, membrane processes, etc., have been used. In this study, three technologies were evaluated for arsenic removal. Results from batch kinetic experiments showed that iron coated sand (IOCS‐2) can remove more than 90 % of As from synthetic water. Experiments were conducted with three different pH values (6, 7, and 8) and an initial As concentration of 260 μg/L. A new material, developed in this study, namely iron coated sponge (IOCSp), was found to have a high capacity in removing both As (V) and As (III). Each gram of IOCSp adsorbed about 160 μg of As within a 9‐hour contact period of IOCSp with As solution. Low pressure nanofiltration removed more than 94 % of As from an influent containing 440 μg/L As. The applied pressure was varied from 85 to 500 kPa.     

Oligoribonuclease is a common downstream target of lithium-induced pAp accumulation in Escherichia coli and human cells

We identified Oligoribonuclease (Orn), an essential Escherichia coli protein and the only exonuclease degrading small ribonucleotides (5mer to 2mer) and its human homologue, small fragment nuclease (Sfn), in a screen for proteins that are potentially regulated by 3′-phosphoadenosine 5′-phosphate (pAp). We show that both enzymes are sensitive to micromolar amounts of pAp in vitro. We also demonstrate that Orn can degrade short DNA oligos in addition to its activity on RNA oligos, similar to what was documented for Sfn. pAp was shown to accumulate as a result of inhibition of the pAp-degrading enzyme by lithium, widely used to treat bipolar disorder, thus its regulatory targets are of significant medical interest. CysQ, the E.coli pAp-phosphatase is strongly inhibited by lithium and calcium in vitro and is a main target of lithium toxicity in vivo. Our findings point to remarkable conservation of the connection between sulfur- and RNA metabolism between E.coli and humans.     

Two biflavonoids from Ouratea nigroviolacea

The leaves of Ouratea nigroviolacea (Ochnaceae) afforded two biflavonoids, ouratine A and B together with agathisflavone and stigmasterol. The biflavonoids were characterized as 4'-O-methylated apigeninyl-(I-6, II-8)-4'-O-methylatedapigenin and 4'-O-methylated apigeninyl-(I-6, II-8) apigenin by spectral and chemical transformation studies.     

Diagnosing benign and malignant lesions in breast tissue sections by using IR-microspectroscopy

The collection of IR spectra through microscope optics and the visualization of the IR data by IR imaging represent a visualization approach, which uses infrared spectral features as a native intrinsic contrast mechanism. To illustrate the potential of this spectroscopic methodology in breast cancer research, we have acquired IR-microspectroscopic data from benign and malignant lesions in breast tissue sections by point microscopy with spot sizes of 30-40 μm. Four classes of distinct breast tissue spectra were defined and stored in the data base: fibroadenoma (a total of 1175 spectra from 14 patients), ductal carcinoma in situ (a total of 1349 spectra from 8 patients), connective tissue (a total of 464 spectra), and adipose tissue (a total of 146 spectra). Artifical neural network analysis, a supervised pattern recognition method, was used to develop an automated classifier to separate the four classes. After training the artifical neural network classifier, infrared spectra of independent external validation data sets (“unknown spectra”) were analyzed. In this way, all spectra (a total of 386) taken from micro areas inside the epithelium of fibroadenomas from 4 patients were correctly classified. Out of the 421 spectra taken from micro areas of the in situ component of invasive ductal carcinomas of 3 patients, 93% were correctly identified. Based on these results, the potential of the IR-microspectroscopic approach for diagnosing breast tissue lesions is discussed.     

PEGylation of protein-based MRI contrast agents improves relaxivities and biocompatibilities

Magnetic resonance imaging (MRI) has emerged as a leading diagnostic technique in clinical and preclinical settings. However, the application of MRI to assess specific disease markers for diagnosis and monitoring drug effect has been severely hampered by the lack of desired contrast agents with high relaxivities, and optimized in vivo retention time. We have reported the development of protein-based MRI contrast agents (ProCA1) by rational design of Gd^3 + binding sites into a stable protein resulting in significantly increased longitudinal (r₁) and transverse (r₂) relaxivities compared to Gd-DTPA. Here, we report a further improvement of protein contrast agents ProCA1 for in vivo imaging by protein modification with various sizes of polyethylene glycol (PEG) chain. PEGylation results in significant increases of both r₁ and r₂ relaxivities (up to 200%), and these high relaxivities persist even at field strengths up to 9.4 T. In addition, our experimental results demonstrate that modified contrast agents have significant improvement of in vivo MR imaging and biocompatibilities including dose efficiency, protein solubility, blood retention time and decreased immunogenicity. Such improvement can be important to the animal imaging and pre-clinical research at high or ultra-high field where there is an urgent need for molecular imaging probes and optimized contrast agent.     

The Armadillo Repeat Gene ZAK IXIK Promotes Arabidopsis Early Embryo and Endosperm Development through a Distinctive Gametophytic Maternal Effect

The proper balance of parental genomic contributions to the fertilized embryo and endosperm is essential for their normal growth and development. The characterization of many gametophytic maternal effect (GME) mutants affecting seed development indicates that there are certain classes of genes with a predominant maternal contribution. We present a detailed analysis of the GME mutant zak ixik (zix), which displays delayed and arrested growth at the earliest stages of embryo and endosperm development. ZIX encodes an Armadillo repeat (Arm) protein highly conserved across eukaryotes. Expression studies revealed that ZIX manifests a GME through preferential maternal expression in the early embryo and endosperm. This parent-of-origin–dependent expression is regulated by neither the histone and DNA methylation nor the DNA demethylation pathways known to regulate some other GME mutants. The ZIX protein is localized in the cytoplasm and nucleus of cells in reproductive tissues and actively dividing root zones. The maternal ZIX allele is required for the maternal expression of MINISEED3. Collectively, our results reveal a reproductive function of plant Arm proteins in promoting early seed growth, which is achieved through a distinct GME of ZIX that involves mechanisms for maternal allele-specific expression that are independent of the well-established pathways.