The U1 snRNP-associated factor Luc7p affects 5′ splice site selection in yeast and human

yLuc7p is an essential subunit of the yeast U1 snRNP and contains two putative zinc fingers. Using RNA–protein cross-linking and directed site-specific proteolysis (DSSP), we have established that the N-terminal zinc finger of yLuc7p contacts the pre-mRNA in the 5′ exon in a region close to the cap. Modifying the pre-mRNA sequence in the region contacted by yLuc7p affects splicing in a yLuc7p-dependent manner indicating that yLuc7p stabilizes U1 snRNP–pre-mRNA interaction, thus reminding of the mode of action of another U1 snRNP component, Nam8p. Database searches identified three putative human yLuc7p homologs (hLuc7A, hLuc7B1 and hLuc7B2). These proteins have an extended C-terminal tail rich in RS and RE residues, a feature characteristic of splicing factors. Consistent with a role in pre-mRNA splicing, hLuc7A localizes in the nucleus and antibodies raised against hLuc7A specifically co-precipitate U1 snRNA from human cell extracts. Interestingly, hLuc7A overexpression affects splicing of a reporter in vivo. Taken together, our data suggest that the formation of a wide network of protein–RNA interactions around the 5′ splice site by U1 snRNP-associated factors contributes to alternative splicing regulation

A Bedside Method for Measuring Effects of a Sedative Drug on Cerebral Function in Newborn Infants

Background: Data on the cerebral effects of analgesic and sedative drugs are needed for the development of safe and effective treatments during neonatal intensive care. Electroencephalography (EEG) is an objective, but interpreter-dependent method for monitoring cortical activity. Quantitative computerized analyses might reveal EEG changes otherwise not detectable. Methods: EEG registrations were retrospectively collected from 21 infants (mean 38.7 gestational weeks; range 27–42) who received dexmedetomidine during neonatal care. The registrations were transformed into computational features and analyzed visually, and with two computational measures quantifying relative and absolute changes in power (range EEG; rEEG) and cortico-cortical synchrony (activation synchrony index; ASI), respectively. Results: The visual assessment did not reveal any drug effects. In rEEG analyses, a negative correlation was found between the baseline and the referential frontal (rho = 0.612, p = 0.006) and parietal (rho = −0.489, p = 0.035) derivations. The change in ASI was negatively correlated to baseline values in the interhemispheric (rho = −0.753; p = 0.001) and frontal comparisons (rho = −0.496; p = 0.038). Conclusion: Cerebral effects of dexmedetomidine as determined by EEG in newborn infants are related to cortical activity prior to DEX administration, indicating that higher brain activity levels (higher rEEG) during baseline links to a more pronounced reduction by DEX. The computational measurements indicate drug effects on both overall cortical activity and cortico-cortical communication. These effects were not evident in visual analysis

A Bedside Method for Measuring Effects of a Sedative Drug on Cerebral Function in Newborn Infants

Background: Data on the cerebral effects of analgesic and sedative drugs are needed for the development of safe and effective treatments during neonatal intensive care. Electroencephalography (EEG) is an objective, but interpreter-dependent method for monitoring cortical activity. Quantitative computerized analyses might reveal EEG changes otherwise not detectable. Methods: EEG registrations were retrospectively collected from 21 infants (mean 38.7 gestational weeks; range 27–42) who received dexmedetomidine during neonatal care. The registrations were transformed into computational features and analyzed visually, and with two computational measures quantifying relative and absolute changes in power (range EEG; rEEG) and cortico-cortical synchrony (activation synchrony index; ASI), respectively. Results: The visual assessment did not reveal any drug effects. In rEEG analyses, a negative correlation was found between the baseline and the referential frontal (rho = 0.612, p = 0.006) and parietal (rho = −0.489, p = 0.035) derivations. The change in ASI was negatively correlated to baseline values in the interhemispheric (rho = −0.753; p = 0.001) and frontal comparisons (rho = −0.496; p = 0.038). Conclusion: Cerebral effects of dexmedetomidine as determined by EEG in newborn infants are related to cortical activity prior to DEX administration, indicating that higher brain activity levels (higher rEEG) during baseline links to a more pronounced reduction by DEX. The computational measurements indicate drug effects on both overall cortical activity and cortico-cortical communication. These effects were not evident in visual analysis

Measuring perceived meaningfulness in day centres for persons with mental illness

Rationale: As support in leading a meaningful and active life, a person with mental illness is often given the opportunity to attend a day centre. However, few studies have investigated the meaningfulness perceived by the person visiting a day centre. For such a purpose, a self-report instrument was developed. Aims: To explore whether perceived meaningfulness, as expressed in the recently developed instrument Evaluation of Perceived Meaning in Day Centers (EPM-DC), could be viewed as one dimension and also to investigate the psychometric properties of this instrument. Methods: Persons with mental illness attending five day centres in Sweden participated and completed the questionnaire. The data were analysed by Rasch analysis. Major findings: The study showed that the concept captured in the instrument could be viewed as unidimensional and the result gave preliminary evidence for sound psychometric properties. Principal conclusion: The results indicate promising signs of validity and reliability, but the suitability of self-reporting may be questioned

Residual DNA analysis in influenza vaccine processing

In cell-based influenza vaccine production, the European Pharmacopoeia demands a host cell residual DNA concentration of less than 10 ng per dose. To reliably measure residual DNA in both process samples and final vaccine using quantitative PCR, DNA preparation prior to analysis is a necessity. Samples from the vaccine purification process contain different buffers, salts, and cell-based compounds, and vary 3–4 logs in DNA concentration from harvest to the final product, which all put strain on the DNA preparation. For accurate determination of DNA concentration, recovery is of high importance. There are many commercially available DNA preparation kits that use different techniques to bind DNA, from spin columns with a DNA-binding membrane or medium (resin) to magnetic beads. However, these kits are mainly developed to purify DNA fragments from gel electrophoresis or genomic DNA from tissues such as blood or cultured cells, and do not have recovery as a priority. Few kits are intended for residual DNA determination in samples with high concentration of a protein or virus product. In this study, prototype media for DNA preparation, in bind-elute and batch mode, were evaluated for recovery, hands-on time, and throughput. In batch mode, recoveries of \u3e 80% were achieved, but the technique exhibited matrix effects on real process samples. In bind-elute mode, recoveries of 40%–60% were achieved after elution. However, recovery could be improved by determination of DNA concentration, while keeping DNA bound to the medium

Haematological toxicity in adult patients receiving craniospinal irradiation - Indication of a dose-bath effect.

The purpose of this study was to investigate the correlation between the haematological toxicity observed in patients treated with craniospinal irradiation, and the dose distribution in normal tissue, specifically the occurrence of large volumes exposed to low dose