Genetic Elements Orchestrating Lactobacillus crispatus Glycogen Metabolism in the Vagina

Glycogen in the female lower reproductive tract is a major carbon source for colonization and acidification by common vaginal Lactobacillus species, such as Lactobacillus crispatus. Previously, we identified the amylopullulanase encoding gene pulA of Lactobacillus crispatus to correlate with the ability to autonomously utilize glycogen for growth. Here, we further characterize genetic variation and differential regulation of pulA affecting the presence of its gene product on the outer surface layer. We show that alpha-glucan degrading activity dissipates when Lactobacillus crispatus is grown on glucose, maltose and maltotriose, in agreement with carbon catabolite repression elements flanking the pulA gene. Proteome analysis of the S-layer confirmed that the amylopullulanase protein is highly abundant in an S-layer enriched fraction, but not in a strain with a defective amylopullulanase variant or in an amylopullulanase-sufficient strain grown on glucose. In addition, we provide evidence that Lactobacillus crispatus pulA mutants are relevant in vivo, as they are commonly observed in metagenome datasets of human vaginal microbial communities. Analysis of the largest publicly available dataset of 1507 human vaginal metagenomes indicates that among the 270 samples that contain a Lactobacillus crispatus pulA gene, 62 samples (23%) had a defective variant of this gene. Taken together, these results demonstrate that both environmental, as well as genetic factors explain the variation of Lactobacillus crispatus alpha-glucosidases in the vaginal environment.     

Trans-Reduction of Cerebral Small Vessel Disease Proteins by Notch-Derived EGF-like Sequences

Cysteine oxidation states of extracellular proteins participate in functional regulation and in disease pathophysiology. In the most common inherited dementia, cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), mutations in NOTCH3 that alter extracellular cysteine number have implicated NOTCH3 cysteine states as potential triggers of cerebral vascular smooth muscle cytopathology. In this report, we describe a novel property of the second EGF-like domain of NOTCH3: its capacity to alter the cysteine redox state of the NOTCH3 ectodomain. Synthetic peptides corresponding to this sequence (NOTCH3 N-terminal fragment 2, NTF2) readily reduce NOTCH3 N-terminal ectodomain polypeptides in a dose- and time-dependent fashion. Furthermore, NTF2 preferentially reduces regional domains of NOTCH3 with the highest intensity against EGF-like domains 12–15. This process requires cysteine residues of NTF2 and is also capable of targeting selected extracellular proteins that include TSP2 and CTSH. CADASIL mutations in NOTCH3 increase susceptibility to NTF2-facilitated reduction and to trans-reduction by NOTCH3 produced in cells. Moreover, NTF2 forms complexes with the NOTCH3 ectodomain, and cleaved NOTCH3 co-localizes with the NOTCH3 ectodomain in cerebral arteries of CADASIL patients. The potential for NTF2 to reduce vascular proteins and the enhanced preference for it to trans-reduce mutant NOTCH3 implicate a role for protein trans-reduction in cerebrovascular pathological states such as CADASIL.     

Halochromic Behavior and Anticancer Effect of New Synthetic Anthocyanidins Complexed with β-Cyclodextrin Derivatives

Anthocyanidins, the aglycons of anthocyanins, are known, beyond their function in plants, also as compounds with a wide range of biological and pharmacological activities, including cytostatic effect against various cancer cells. The nature and position of the substituents in the flavylium cation is essential for such biological properties, as well as the equilibrium between the multistate of the different chemical species that are generated by the flavylium cation, including quinoidal base, hemiketal, and cis- and trans-chalcones. In this work, eight new flavylium derivatives were synthesized, characterized for confirmation of the structure by FT-IR and 2D-NMR, and investigated in vitro as possible cytostatic compounds against HCT116 and HepG2 cancer cells. The most active two compounds were explored for their halochromic properties that can influence the biological activity and subjected to molecular encapsulation in β-cyclodextrin derivatives in order to increase their solubility in water and bioavailability. The anticancer effect was influenced by the position (6-, 7-, or 8-) of the methoxy group in the β-ring of the methoxy-4′-hydroxy-3′-methoxyflavylium cation, while the study of the halochromic properties revealed the important role played by the chalcone species of the pH-dependent multistate in both the uncomplexed and inclusion complex forms of these anthocyanidins.     

Performance analysis of Viterbi decoding for 64-DAPSK and 64-QAMmodulated OFDM signals

The multilevel modulation techniques of 64-quadrature amplitude modulation (QAM) and 64-differential amplitude and phase-shift keying (DAPSK) have been proposed in combination with the orthogonal frequency-division multiplexing (OFDM) scheme for digital terrestrial video broadcasting (DTVB). With this system a data rate of 34 Mb/s can be transmitted over an 8-MHz radio channel. A comparison of these modulation methods in the uncoded case has been presented by Engels and Rohling (see European Trans. Telecommun., vol.6, p.633-40, 1995). The channel coding scheme proposed for DTVB by Schafer (see Proc. Int. Broadcasting Convention, Amsterdam, The Netherlands, p.79-84, 1995) consists of an inner convolutional code concatenated with an outer Reed-Solomon (RS) code. In this paper the performance of the convolutional codes is analyzed for the two modulation techniques. This analysis includes soft decision Viterbi (1971) decoding of the convolutional code. For soft decision decoding of DAPSK modulated signals a new metric is developed     

Identifying Strategies to Support Junior Secondary Students to Engage in Scientific Investigation Tasks

Abstract

This article describes the outcome of a collaborative project between the Hong Kong Institute of Education and four secondary schools that aims to promote the development of scientific investigation skills. The project team designed scientific investigation tasks collaboratively with the teachers and provided school-based support when the tasks were implemented. A total of six teachers and 575 students were involved. Data were collected through questionnaires completed by the students and individual interviews with science teachers about their classroom practice after the completion of the project. The findings suggest that the students did not meet many difficulties and that there were positive influences on students’ interest in learning science. The teachers perceived that there were challenges related to raising students’ self-regulated learning abilities, structuring tasks that were at appropriate levels of difficulty, and promoting group cooperation among the students. Finally, the article argues that the strategies implemented in this study were effective, though it takes much time and effort to help students develop self-regulated learning abilities. The conclusion suggests that teachers consider these challenges collectively and proposes a two-staged model for planning scientific investigation tasks.     

Targeted reconstruction with differential planar computed tomography

In computed tomography, targeted (or zoomed) reconstruction is usually used to achieve high-resolution reconstruction to a small region-of-interest (ROI). This paper reports a novel targeted reconstruction method based on the recently developed differential planar computed tomography technology. With the central-slice sinogram of scanning a planar object, the primary orientation of the object is first determined for the first projection. Then by identifying the dimensions of the ROI and its offsets to the centre-of-rotation (COR), the targeted reconstruction matrix is defined, which just covers the ROI and follows its initial orientation at the scan start time. By scanning and reconstructing a small planar ROI soldered on a large planar substrate, we demonstrate the advantages of the proposed method over traditional CT, including the smaller reconstruction size and less computation time, high-resolution reconstruction and good-orientated CT image for subsequent de-layering and visualization.