Intracellular serine protease 1 of Bacillus subtilis is formed in vivo as an unprocessed, active protease in stationary cells.

Western immunoblots and assays of Bacillus subtilis extracts showed that intracellular serine protease 1 is produced in a form larger than previously reported, appears not to have undergone N-terminal processing, and is active in the presence or absence of calcium. No evidence for an inactive precursor form of the protease was found.     

Solid-state fermentation of sweet sorghum to ethanol

Solid-state fermentation of chopped sweet sorghum particles to ethanol was studied in static flasks using an ethanol tolerant yeast strain. The influence of various process parameters, such as temperature, yeast cell concentration, and moisture content, on the rate and extent of ethanol fermentation was investigated. Optimal values of these parameters were found to be 35 degrees C, 7 x 10(8) cells/g raw sorghum, and 70% moisture level, respectively.     

Experiments on the reproductibility of results from EUF soil extracts with possible improvements resulting from these experiments

Summary The reproducibility of EUF-Ca, K, Na, P, B, NO₃–N and EUF extractable N for fraction 1 (200 V, 20° C, 30 minutes) was within a CV range of 4.54–12.28%.The summation of results for fraction 1 (200 V, 20°C, 30 minutes) and fraction 2 (400 V, 80°C, 5 minutes) gave an improved CV range of 4.23–8.81%.EUF-Mg obtained from fraction 1 (200 V, 20°C, 30 minutes), fraction 2 (400 V, 80°C, 5 minutes) and the acid fraction gave a CV of 6.81%.Results from the acid fraction gave poor reproducibility for Zn, (CV=45.1%). The CV of 12.5% for Mn is low compared to the CV of 30–40% usually obtained.The recovery of Zn, Mn and Mg from the acid fraction was improved. The reproducibility of Zn and Mg was also improved by (a) reducing the sources of contamination; (b) introducing a second filter at the cathode; (c) steeping the filters for 48 hours.     

Objective assessment of treatment response in hirsutism

A simple photographic technique was used to assess the objective response of hirsute women to treatment with cyproterone acetate (CA). The skin in front of the left ear was shaved and photographs taken immediately and after 1 week, and hair growth per week estimated by averaging the length of 20 hairs in the magnified photographs. The precision, repeatability and patient acceptance of the method were found to be satisfactory. Basal hair growth rates were 2.28 +/- 0.4 mm/week (mean +/- SD, n = 34) and showed a significant correlation with hirsutism scores derived from a standard physician-rated scale. During therapy with CA the mean (+/- SD) improvement in hair growth rate was 19 +/- 13%, whereas physician-rated hirsutism scores improved by 33 +/- 20%. The reduction in hair growth rate showed no significant correlation with improvement assessed using subjective rating by either physician or patient. The greater improvement in physician-rated scores compared with hair growth rate assessment suggests that hair shaft width, colour and other factors may be as important as hair length when assessing treatment response.     

Deaths of drug addicts in the United Kingdom 1967-81.

A search of the Home Office index of notified drug addicts identified 1499 deaths during 1967-81, of which 226 (15%) were of therapeutic addicts--that is, patients who had become addicted during medical treatment with a notifiable drug--and 1273 (85%) were of non-therapeutic addicts. The crude mortality fell from 23.5/1000/year for the period 1968-70 to 18.4/1000/year for 1978-80. Altogether 416 addicts aged under 50 at notification died after 1972, which was 16 times the number of deaths expected in a population with a similar age and sex composition. A more detailed examination of the cohorts of addicts notified each year showed little variation between them in the first two years of follow up. Nineteen addicts (1.6%) had died by 31 December of the year of their notification and 39 (3.3%) one year later. These figures may be an indication of the clinical course of addiction. Most deaths of non-therapeutic addicts in which a drug was implicated (939 cases (74%] were due to medically prescribed drugs--barbiturates at first and later opiates such as dipipanone hydrochloride and dextromoramide. Heroin was implicated in only 65 (7%) of these deaths. The Home Office index is a valuable source for identifying drugs of abuse and serious problems of addiction. The fact that prescribed drugs are causing the death of so many addicts demands a response from the medical profession.     

Apparent resilience of a declining Afro‐Palaearctic migrant to forest loss on the wintering grounds

Populations of long‐distance migrant birds are declining but it is unknown what role land cover change in non‐breeding areas may be playing in this process. Using compositional analysis, we assessed habitat selection by one such migrant, the Wood Warbler Phylloscopus sibilatrix, at a wintering site in the forest–savannah transition zone in Eastern Region, Ghana. There was a preference for forest, a habitat that is in marked decline at this site. Annual habitat mapping revealed that the area of forest declined by 26% between 2011/12 and 2013/14, mainly through clearance for conversion to arable land. Numbers of birds changed throughout the season, but despite the reduction in the preferred forest habitat, there was no change in the total number of birds recorded at the site over the study period. The number of birds recorded at a point was positively related to the proportion of cleared land, plantation and, to a lesser extent, dense forest within 100 m. Investigation of the fine‐scale habitat preferences of radiotagged Wood Warblers suggested that there was an optimum number of trees, around 66–143 per hectare, at which estimated probability of occupancy was 0.5, falling to a probability of 0.2 at 25 trees per hectare. We suggest that Wood Warblers may be buffered against the loss of forest habitat by their ability to utilize degraded habitats, such as well‐wooded farmland, that still retain a substantial number of trees. However, the continued loss of trees, from both forest and farmland is ultimately likely to have a negative impact on wintering Wood Warblers in the long‐term.     

Habitat use and tree selection of a declining Afro-Palaearctic migrant at sub-Saharan staging and wintering sites

Capsule: Wood Warblers Phylloscopus sibilatrix showed significant selection for tree species and woodland characteristics at staging and wintering sites in sub-Saharan Africa.

Aims: To investigate home range size, habitat and tree species selection of Wood Warblers at a staging site in Burkina Faso (Koubri) and a wintering site in Ghana (Pepease).

Methods: Comparing habitat recorded at locations of radio-tagged birds and at control points, we investigated whether there was habitat and tree species selection. We also compared home range size of individual birds between the two sites.

Results: Home range size did not differ between the two sites. There was significant selection for tree species at both Koubri and Pepease: Anogeissus leiocarpus and Albizia zygia, respectively. At Koubri, there was significant avoidance of the most common tree species (Azadirachta indica, Mangifera indica (both non-native), Vitellaria paradoxa and Acacia spp.). In addition, there was a preference for taller trees and greater tree density at both sites. However, the probability of a point being used declined with increasing number of taller (>14?m) trees.

Conclusion: Fine-scale selection of woodland habitats suggests that Wood Warblers are likely to suffer the consequences of ongoing land-use change in their West African wintering grounds.     

Hydrophobized triphenyl phosphonium derivatives for the preparation of mitochondriotropic liposomes: choice of hydrophobic anchor influences cytotoxicity but not mitochondriotropic effect

Context: Nanocarrier-based strategies to achieve delivery of bioactives specifically to the mitochondria are being increasingly explored due to the importance of mitochondria in critical cellular processes.

Objective: To test the ability of liposomes modified with newly synthesized triphenylphosphonium (TPP)–phospholipid conjugates and to test their use in overcoming the cytotoxicity of stearyl triphenylphosphonium (STPP)-modified liposomes when used for delivery of therapeutic molecules to the mitochondria.

Methods: TPP–phospholipid conjugates with the dioleoyl, dimyristoyl or dipalmitoyl lipid moieties were synthesized and liposomes were prepared with these conjugates in a 1?mol% ratio. The subcellular distribution of the liposomes was tested by confocal microscopy. Furthermore, the liposomes were tested for their effect on cell viability using a MTS assay, on cell membrane integrity using a lactate dehydrogenase assay and on mitochondrial membrane integrity using a modified JC-1 assay.

Results: The liposomes modified with the new TPP–phospholipid conjugates exhibited similar mitochondriotropism as STPP-liposomes but they were more biocompatible as compared to the STPP liposomes. While the STPP-liposomes had a destabilizing effect on cell and mitochondrial membranes, the liposomes modified with the TPP–phospholipid conjugates did not demonstrate any such effect on biomembranes.

Conclusions: Using phospholipid anchors in the synthesis of TPP–lipid conjugates can provide liposomes that exhibit the same mitochondrial targeting ability as STPP but with much higher biocompatibility.     

Engineering of CRISPR/Cas9‐mediated potyvirus resistance in transgene‐free Arabidopsis plants

Members of the eukaryotic translation initiation factor (eIF) gene family, including eIF4E and its paralogue eIF(iso)4E, have previously been identified as recessive resistance alleles against various potyviruses in a range of different hosts. However, the identification and introgression of these alleles into important crop species is often limited. In this study, we utilise CRISPR/Cas9 technology to introduce sequence‐specific deleterious point mutations at the eIF(iso)4E locus in Arabidopsis thaliana to successfully engineer complete resistance to Turnip mosaic virus (TuMV), a major pathogen in field‐grown vegetable crops. By segregating the induced mutation from the CRISPR/Cas9 transgene, we outline a framework for the production of heritable, homozygous mutations in the transgene‐free T₂ generation in self‐pollinating species. Analysis of dry weights and flowering times for four independent T₃ lines revealed no differences from wild‐type plants under standard growth conditions, suggesting that homozygous mutations in eIF(iso)4E do not affect plant vigour. Thus, the established CRISPR/Cas9 technology provides a new approach for the generation of Potyvirus resistance alleles in important crops without the use of persistent transgenes.