Targeted knock-down of miR21 primary transcripts using snoMEN vectors induces apoptosis in human cancer cell lines

We have previously reported an antisense technology, 'snoMEN vectors', for targeted knock-down of protein coding mRNAs using human snoRNAs manipulated to contain short regions of sequence complementarity with the mRNA target. Here we characterise the use of snoMEN vectors to target the knock-down of micro RNA primary transcripts. We document the specific knock-down of miR21 in HeLa cells using plasmid vectors expressing miR21-targeted snoMEN RNAs and show this induces apoptosis. Knock-down is dependent on the presence of complementary sequences in the snoMEN vector and the induction of apoptosis can be suppressed by over-expression of miR21. Furthermore, we have also developed lentiviral vectors for delivery of snoMEN RNAs and show this increases the efficiency of vector transduction in many human cell lines that are difficult to transfect with plasmid vectors. Transduction of lentiviral vectors expressing snoMEN targeted to pri-miR21 induces apoptosis in human lung adenocarcinoma cells, which express high levels of miR21, but not in human primary cells. We show that snoMEN-mediated suppression of miRNA expression is prevented by siRNA knock-down of Ago2, but not by knock-down of Ago1 or Upf1. snoMEN RNAs colocalise with Ago2 in cell nuclei and nucleoli and can be co-immunoprecipitated from nuclear extracts by antibodies specific for Ago2

Stereochemistry of Bistricyclic Aromatic Enes and Related Polycyclic Systems

Bistricyclic aromatic enes (BAEs) and related polycyclic systems are a class of molecular materials that display a rich variety of conformations, dynamic stereochemistry and switchable chirality, color, and spectroscopic properties. This is due to the a subtle interplay of the inherent preference for planarity of aromatic systems and the competing necessity of non-planarity due to intramolecular overcrowding in the fjord regions built into the general molecular structure of BAEs. The conformational, dynamic, and spectroscopic properties may be designed and fine-tuned, e.g., by variation of the bridging groups X and Y, the overcrowding in the fjord regions, extensions of the aromatic system, or other modifications of the general BAE structure, based on the fundamental understanding of the structure-property relationships (SPR). The present review provides an analysis of the conformational spaces and the dynamic stereochemistry of overcrowded bistricyclic aromatic enes applying fundamental symmetry considerations. The symmetry analysis presented here allows deeper insight into the conformations, chirality, and the mechanisms of the dynamic stereochemistry, and will be instrumental in future computational studies

The Hyperactivity/Inattention Subscale of the Strengths and Difficulties Questionnaire Predicts Short- and Long-Term Weight Loss in Overweight Children and Adolescents Treated as Outpatients

Objective: The success of treatment for pediatric obesity is variable and often unsatisfactory. This study elucidates the influence of inattention and hyperactivity/impulsivity on short- and long-term weight loss and maintenance after outpatient treatment. Methods: We included 8- to 16-year-old overweight and obese participants treated in 17 multidisciplinary outpatient treatment centers in a nationwide observational study. All treatment centers that reported long-term (1-year) follow-up weight data of at least 60% of the participants were included. At the beginning and end of treatment and at 1 year follow-up weight and height were measured at the center. Inattention and hyperactivity/impulsivity were assessed with the hyperactivity/inattention subscale (HI) of the parent-rated Strengths and Difficulty Questionnaire (SDQ). General linear models were used with the standard deviation scores of the BMI (BMI-SDS) as dependent variable and HI scores as main independent variable adjusting for age, sex, baseline BMI-SDS, and center. Results: 394 participants were included (57% female, age: 11.7± 2.0 years, baseline BMI-SDS 2.32 ±.46 kg/m2). HI scores were significantly associated with short- and long-term BMI-SDS (p < 0.0005), with higher baseline HI scores predicting less weight loss. Conclusions: Our results indicate that inattention and hyperactivity/impulsivity are linearly associated with reduced short- and long-term weight loss. Implications for treatment are discussed