Fluorescence Correlation Spectroscopy Reveals Efficient Cytosolic Delivery of Protein Cargo by Cell-Permeant Miniature Proteins.

New methods for delivering proteins into the cytosol of mammalian cells are being reported at a rapid pace. Differentiating between these methods in a quantitative manner is difficult, however, as most assays for evaluating cytosolic protein delivery are qualitative and indirect and thus often misleading. Here we make use of fluorescence correlation spectroscopy (FCS) to determine with precision and accuracy the relative efficiencies with which seven different previously reported "cell-penetrating peptides" (CPPs) transport a model protein cargo-the self-labeling enzyme SNAP-tag-beyond endosomal membranes and into the cytosol. Using FCS, we discovered that the miniature protein ZF5.3 is an exceptional vehicle for delivering SNAP-tag to the cytosol. When delivered by ZF5.3, SNAP-tag can achieve a cytosolic concentration as high as 250 nM, generally at least 2-fold and as much as 6-fold higher than any other CPP evaluated. Additionally, we show that ZF5.3 can be fused to a second enzyme cargo-the engineered peroxidase APEX2-and reliably delivers the active enzyme to the cell interior. As FCS allows one to realistically assess the relative merits of protein transduction domains, we anticipate that it will greatly accelerate the identification, evaluation, and optimization of strategies to deliver large, intact proteins to intracellular locales

‘It’s a different world out there’: Improving how academics prepare health science students for rural and Indigenous practice in Australia.

Rural and Aboriginal and Torres Strait Islander (Indigenous) health content in undergraduate health science curricula in Western Australia has been limited. In 2008, a three-and-a-half-day, rurally-based, intercultural and inter-disciplinary programme for academics from three universities aimed to improve how academics prepared health science students for work in this area. Situated learning theory underpinned the programme's design, which prioritised context and participation in the construction of knowledge: academics lived ‘on country’ and participated in the lived experience of a rural and Indigenous community. Semi-structured phone interviews with 21 academics four months later indicated this approach had radically changed thinking and led to a desire to improve rural and Indigenous health and teaching practice. Targeting academics to learn about rural and Indigenous health in situ is one promising strategy for improving undergraduate health science education in this priority area

Examination of Fluconazole-Induced Alopecia in an Animal Model and Human Cohort.

Fluconazole-induced alopecia is a significant problem for patients receiving long-term therapy. We evaluated the hair cycle changes of fluconazole in a rat model and investigated potential molecular mechanisms. Plasma and tissue levels of retinoic acid were not found to be causal. Human patients with alopecia attributed to fluconazole also underwent detailed assessment and in both our murine model and human cohort fluconazole induced telogen effluvium. Future work further examining the mechanism of fluconazole-induced alopecia should be undertaken

High-resolution Near-Infrared Images and Models of the Circumstellar Disk in HH 30

We present Hubble Space Telescope (HST) Near-Infrared Camera and Multi-object Spectrometer (NICMOS) observations of the reflection nebulosity associated with the T Tauri star HH 30. The images show the scattered light pattern characteristic of a highly inclined, optically thick disk with a prominent dustlane whose width decreases with increasing wavelength. The reflected nebulosity exhibits a lateral asymmetry in the upper lobe on the opposite side to that reported in previously published Wide Field Planetary Camera 2 (WFPC2) images. The radiation transfer model which most closely reproduces the data has a flared accretion disk with dust grains larger than standard interstellar medium grains by a factor of approximately 2.1. A single hotspot on the stellar surface provides the necessary asymmetry to fit the images and is consistent with previous modeling of the light curve and images. Photometric analysis results in an estimated extinction of Av>~80; however, since the photometry measures only scattered light rather than direct stellar flux, this a lower limit. The radiative transfer models require an extinction of Av = 7,900.Comment: Accepted for publication in Ap.

Clostridium difficile Infection in Outpatients, Maryland and Connecticut, USA, 2002–2007

Clostridium difficile, the most commonly recognized diarrheagenic pathogen among hospitalized persons, can cause outpatient diarrhea. Of 1,091 outpatients with diarrhea, we found 43 (3.9%) who were positive for C. difficile toxin. Only 7 had no recognized risk factors, and 3 had neither risk factors nor co-infection with another enteric pathogen