The isolation of DNA from agarose gels by electrophoretic elution onto malachite green-polyacrylamide columns

Electrophoretic elution of DNA coupled with direct adsorption onto malachite green-polyacrylamide columns was used to isolate double- and single-stranded DNA from agarose gels. Subsequently, DNA was eluted with a high salt buffer and filtered through Sephadex which permitted recovery of the DNA in a low salt buffer at concentrations suitable for heteroduplex analysis by electron microscopy. This method was tested by examining hetero-duplexes formed from the isolated complementary single strands of T7 wild type DNA and a T7 deletion mutant. More than 80% of the reannealed molecules were intact heteroduplexes showing the deletion loop. Irradiation of single-stranded DNA with 254 nm light resulted in distorted, convoluted heteroduplexes while 366 nm light did not show this effect.     

Orientation of pigments and pigment-protein complexes in the green photosynthetic bacterium Prosthecochloris aestuarii

The orientation of pigments and pigment-protein complexes of the green photosynthetic bacterium Prosthecochloris aestuarii was studied by measurement of linear dichroism spectra at 295 and 100 K. Orientation of intact cells and membrane vesicles (Complex I) was obtained by drying on a glass plate. The photochemically active pigment-protein complexes (photosystem-protein complex and reaction center pigment-protein complex) and the antenna bacteriochlorophyll a protein were oriented by pressing a polyacrylamide gel. The data indicate that the near-infrared transitions (Q_y) of bacteriochlorophyll c and most bacteriochlorophyll a molecules have a relatively parallel orientation to the membrane, whereas the Q_y transitions of the bacteriochlorophyll a in the antenna protein are oriented predominantly perpendicularly to the membrane. Carotenoids and the Q_x transitions (590–620 nm) of bacteriochlorophyll a, not belonging to the bacteriochlorophyll a protein, have a relatively perpendicular orientation to the membrane. The absorption and linear dichroism spectra indicate the existence of different pools of bacteriochlorophyll c in the chlorosomes and of carotenoid and bacteriopheophytin c in the cell membrane. The results suggest that the photosystem-protein and reaction center pigment-protein complexes are oriented with their short axes approximately perpendicular to the plane of the membrane. The symmetry axis of the bacteriochlorophyll a protein has an approximately perpendicular orientation.     

Sequence homologies between Escherichia coli and chloroplast ribosomal DNA as seen by heteroduplex analysis

Circular Vicia faba (broad bean) chloroplast DNA was hybridized to the restriction fragment BamHI B from the DNA of the transducing phage lambda rif^d18, which carries the Escherichia coli ribosomal RNA operon rrnB. Cytochrome spreadings of the heteroduplexes show homologies in the 16 S and 23 S rRNA regions, but none in the spacer. The same lambda rif^d18 fragment was hybridized to the Vicia cpDNA ²SalI fragment 3, which contains the Vicia rBNA operon, resulting in an analogous heteroduplex configuration. Cytochrome spreadings of this heteroduplex in increasing concentrations of formamide reveal regions of incomplete homologies. Heteroduplexes between the E. coli rrnD operon, obtained from the recombinant plasmid pBK8, and circular Vicia cpDNA revealed homologies in the spacer region as well as in the 16 S and 23 S rRNA region. Hybrids between all three types of rDNA and their homologous rRNAs were prepared using the mica adsorption technique. They show that the 23 S, 16 S, and 5 S rRNAs are transcribed from the same strand of Vicia cpDNA. The positions of the rRNAs were measured and compared to the heteroduplex structure. It was observed that the E. coli rrnD operon in the plasmid pBK8 contains two 5 S rRNA sequences near the distal end.     

Fluorescence emission spectra of chloroplasts and subchloroplast preparations at low temperature.

A study was made of the chlorophyll fluorescence spectra between 100 and 4.2 K of chloroplasts of various species of higher plants (wild strains and chlorophyll b mutants) and of subchloroplast particles enriched in Photosystem I or II. The chloroplast spectra showed the well known emission bands at about 685, 695 and 715--740 nm; the System I and II particles showed bands at about 675, 695 and 720 nm and near 685 nm, respectively. The effect of temperature lowering was similar for chloroplasts and subchloroplast particles; for the long wave bands an increase in intensity occurred mainly between 100 and 50 K, whereas the bands near 685 nm showed a considerable increase in the region of 50--4.2 K. In addition to this we observed an emission band near 680 nm in chloroplasts, the amplitude of which was less dependent on temperature. The band was missing in barley mutant no. 2, which lacks the light-harvesting chlorophyll a/b-protein complex. At 4.7 K the spectra of the variable fluorescence (Fv) consisted mainly of the emission bands near 685 and 695 nm, and showed only little far-red emission and no contribution of the band at 680 nm. From these and other data it is concluded that the emission at 680 nm is due to the light-harvesting complex, and that the bands at 685 and 695 nm are emitted by the System II pigment-protein complex. At 4.2 K, energy transfer from System II to the light-harvesting complex is blocked, but not from the light-harvesting to the System I and System II complexes. The fluorescence yield of the chlorophyll species emitting at 685 nm appears to be directly modulated by the trapping state of the reaction center.     

Health Care Utilization and Symptom Severity in Ghanaian Children – a Cross-Sectional Study

The aim of this study was to identify factors influencing health care utilization behavior for children with mild or severe disease symptoms in rural Ghana. Between March and September 2008 a cross-sectional health care utilization survey was conducted and 8,715 caregivers were interviewed regarding their intended behavior in case their children had mild or severe fever or diarrhea. To show associations between hospital attendance and further independent factors (e.g. travel distance or socio-economic status) prevalence ratios were calculated for the four disease symptoms. A Poisson regression model was used to control for potential confounding. Frequency of hospital attendance decreased constantly with increasing distance to the health facility. Being enrolled in the national health insurance scheme increased the intention to attend a hospital. The effect of the other factors diminished in the Poisson regression if modeled together with travel distance. The observed associations weakened with increasing severity of symptoms, which indicates that barriers to visit a hospital are less important if children experience a more serious illness. As shown in other studies, travel distance to a health care provider had the strongest effect on health care utilization. Studies to identify local barriers to access health care services are important to inform health policy making as they identify deprived populations with low access to health services and to early treatment.     

Higher Risk Perception of HIV than of Chlamydia and HPV among Secondary School Students in Two German Cities

Background

Chlamydia and genital human papillomavirus (HPV) are the two most common sexually transmitted infections (STIs) among teens and young adults in industrialised countries. The majority of adolescents, however, have limited or no knowledge of these infections. Within the context of a cross-sectional survey on awareness and knowledge of sexually transmitted infections, secondary school students attending the 8th grade and above in Bremen and Bremerhaven, two cities in northern Germany, were asked to rate the risk of peers to get infected with HIV, HPV or chlamydia.

Methods

Between October and December 2011, students aged 12–20 years completed an anonymous, self-administered questionnaire at their school. In addition to answering questions on awareness and knowledge of sexually transmitted infections, all students were also asked to rate the risk of peers to get infected with HIV, HPV or chlamydia. Furthermore, those reporting ever having sexual intercourse were asked to rate their own risk of getting infected with each of the three infections.

Results

1,148 students, 55% female, completed the questionnaire. 27% of the students reported having had sexual intercourse. 68% of all students rated the risk of same-aged students to get infected with HIV/AIDS as high/medium. The corresponding proportions for HPV and chlamydia were 19 and 25% respectively. Those reporting ever having sexual intercourse generally perceived their own risk of getting infected with HIV, chlamydia or HPV as lower than that of their peers.

Conclusion

Generally, the risk of getting infected with HIV was perceived as being higher than that of getting infected with HPV or chlamydia, most likely due to the fact that the students were more aware of HIV than of the other two infections. Efforts should be made to improve awareness and knowledge of HPV and chlamydia among school going adolescents, and to make them realize that these are common infections that are preventable.     

A Clinical Evaluation of Statin Pleiotropy: Statins Selectively and Dose-Dependently Reduce Vascular Inflammation

Statins are thought to reduce vascular inflammation through lipid independent mechanisms. Evaluation of such an effect in atherosclerotic disease is complicated by simultaneous effects on lipid metabolism. Abdominal aortic aneurysms (AAA) are part of the atherosclerotic spectrum of diseases. Unlike atherosclerotic occlusive disease, AAA is not lipid driven, thus allowing direct evaluation of putative anti-inflammatory effects. The anti-inflammatory potency of increasing doses (0, 20 or 40 mg/day) simvastatin or atorvastatin was evaluated in 63 patients that were at least 6 weeks on statin therapy and who underwent open AAA repair. A comprehensive analysis using immunohistochemistry, mRNA and protein analyses was applied on aortic wall samples collected during surgery. The effect of statins on AAA growth was analyzed in a separate prospective study in incorporating 142 patients. Both statins equally effectively and dose-dependently reduced aortic wall expression of NFκB regulated mediators (i.e. IL-6 (P<0.001) and MCP-1 (P<0.001)); shifted macrophage polarization towards a M2 phenotype (P<0.0003); selectively reduced macrophage-related markers such as cathepsin K and S (P<0.009 and 0.0027 respectively), and ALOX5 (P<0.0009), and reduced vascular wall NFκB activity (40 mg/day group, P<0.016). No effect was found on other cell types. Evaluation of the clinical efficacy of statins to reduce AAA progression did not indicate an effect of statins on aneurysm growth (P<0.337). Hence, in the context of AAA the clinical relevance of statins pleiotropy appears minimal.     

Das essenzielle Spurenelement Zink. Ein Metall in Biologie und Medizin

The essential trace element zinc The human body contains 2–3 grams of the essential trace element zinc, and uptake as well as excretion underlies a tight control. If this balance is disturbed, a number of biological processes are affected: among other tasks, zinc is important for the development and function of the central nervous system, the immune defense, and the production and function of insulin. This is based on over 300 different zinc‐containing enzymes, several other zinc proteins, and a role of zinc in intracellular signal transduction.