A technique to detect and to quantify fasciocutaneous blood vessels in small laboratory animals ex vivo

PURPOSE: A microangiographical technique is described, which allows visualization of small and capillary blood vessels and quantification of fasciocutaneous blood vessels by means of digital computer analysis in very small laboratory animals. MATERIALS AND METHODS: The left carotid artery of 20 nu/nu mice was cannulated (26 gauge) and a mixture of gelatin, bariumsulfate, and green ink was injected according to standardized protocol. Fasciocutaneous blood vessels were visualized by digital mammography and analyzed for vessel length and vessel surface area as standardized units [SU] by computer program. RESULTS: With the described microangiography method, fasciocutaneous blood vessels down to capillary size level can be clearly visualized. Regions of interest (ROIs) can be defined and the containing vascular network quantified. Comparable results may be obtained by calculating the microvascular area index (MAI) and the microvascular length index (MLI), related to the ROIs size. Identical ROIs showed a high reproducibility for measured [SU] < 0.01 +/- 0.0012%. CONCLUSION: Combining microsurgical techniques, pharmacological knowledge, and modern digital image technology, we were able to visualize small and capillary blood vessels even in small laboratory animals. By using our own computer analytical program, quantification of vessels was reliable, highly reproducible, and fast.

Characterization of the fetuin-binding fraction of Neospora caninum tachyzoites and its potential involvement in host-parasite interactions

Terminal sialic acid residues on surface-associated glycoconjugates mediate host cell interactions of many pathogens. Addition of sialic acid-rich fetuin enhanced, and the presence of the sialidiase inhibitor 2-deoxy-2,3-dehydro-N-acetylneuraminic acid reduced, the physical interaction of Neospora caninum tachyzoites and bradyzoites with Vero cell monolayers. Thus, Neospora extracts were subjected to fetuin-agarose affinity chromatography in order to isolate components potentially interacting with sialic acid residues. SDS-PAGE and silver staining of the fetuin binding fraction revealed the presence of a single protein band of approximately 65 kDa, subsequently named NcFBP (Neospora caninum fetuin-binding protein), which was localized at the apical tip of the tachyzoites and was continuously released into the surrounding medium in a temperature-independent manner. NcFBP readily interacted with Vero cells and bound to chondroitin sulfate A and C, and anti-NcFBP antibodies interfered in tachyzoite adhesion to host cell monolayers. In additon, analysis of the fetuin binding fraction by gelatin substrate zymography was performed, and demonstrated the presence of two bands of 96 and 140 kDa exhibiting metalloprotease-activity. The metalloprotease activity readily degraded glycosylated proteins such as fetuin and bovine immunoglobulin G heavy chain, whereas non-glycosylated proteins such as bovine serum albumin and immunoglobulin G light chain were not affected. These findings suggest that the fetuin-binding fraction of Neospora caninum tachyzoites contains components that could be potentially involved in host-parasite interactions.

Gunshot-related displacement of skin particles and bacteria from the exit region back into the bullet path

In previous studies, it was shown that there is a gunshot-related transport of skin particles and microorganisms from the entrance region into the depth of the bullet path. The present study deals with the question of whether gunshots may also cause a retrograde transport of skin particles and microorganisms from the bullet exit region back into the bullet path. For this purpose, we used a composite model consisting of rectangular gelatin blocks and pig skin. The skin pieces were firmly attached to the gelatin blocks on the side where the bullet was to exit. Prior to the test shots, the outer surface of the pig skin was contaminated with a thin layer of a defined bacterial suspension. After drying the skin, test shots were fired from a distance of 10 m using cartridges calibre .38 spec. with different bullet types. Subsequent analyses showed that in all shots with full penetration of the composite model, the bullet path contained displaced skin particles and microorganisms from the skin surface at the exit site. These could be regularly detected in the distal 6-8 cm of the track, occasionally up to a distance of 18 cm from the exit hole. The distribution of skin particles and microorganisms is presented and the possible mechanism of this retrograde transport is discussed.

Heikinpäivä in Hancock

Each January in Hancock, Michigan, the Heikinpäivä Midwinter Festival offers local residents three weeks of activities highlighting the continuing role Finnish culture has in the area. Utilizing a set of fading Finnish midwinter traditions surrounding the day of Heikki, or Henrik, this festival has grown from a brief day-long gathering to a long period of activity incorporating films, craft and cooking classes, religious services, and more traditional festival events such as a parade, games, feasting, music, and dancing. This festival has complex origins in more commonplace agricultural traditions brought from Finland by immigrants, which are often no longer commonly remembered in Finland to this day. In this paper, I will examine the complex history of this festival both through its Finnish origins and through its current incarnation in Michigan. Through this festival, we can see the role Finnish heritage has as a simultaneous marker of cultural pride and deprecation. The place Finnish heritage has as a tool in community and economic development in the City of Hancock and the wider region will also be explored. Finally, the function of the festival as a means of maintaining traditions seemingly doomed to fade with time will also be explored.

Performance characteristics and design recommendations for biomass-burning stoves using earthen construction materials

This report provides an analysis of the thermal performance and emissions characteristics of improved biomass stoves constructed using earthen materials. Commonly referred to as mud stoves, this type of improved stove incorporates high clay content soil with an organic binder in the construction of its combustion chamber and body. When large quantities of the mud material are used to construct the stove body, the stove does not offer significant improvements in fuel economy or air quality relative to traditional open fire cooking. This is partly because a significant amount of heat is absorbed by the mass of the stove reducing combustion efficiency and heat transfer to the cook pot. An analysis of the thermal and mechanical properties of stove materials was also performed. A material mixture containing a one‐to‐one ratio by volume of high content clay soil and straw was found to have thermal properties comparable to fired ceramics used in more advanced improved stove designs. Feedback from mud stove users in Mauritania and Mali, West Africa was also collected during implementation. Suggestions for stove design improvements were developed based on this information and the data collected in the performance, emissions, and material properties analysis. Design suggestions include reducing stove height to accommodate user cooking preferences and limiting overall stove mass to reduce heat loss to the stove body.

Activation of the epidermal growth factor receptor (EGFR) by a novel metalloprotease pathway

Neutrophil Elastase (NE) is a pro-inflammatory protease present at higher than normal levels in the lung during inflammatory disease. NE regulates IL-8 production from airway epithelial cells and can activate both EGFR and TLR4. TACE/ADAM17 has been reported to trans-activate EGFR in response to NE. Here, using 16HBE14o-human bronchial epithelial cells we demonstrate a new mechanism by which NE regulates both of these events. A high molecular weight soluble metalloprotease activity detectable only in supernatants from NE-treated cells by gelatin and casein zymography was confirmed to be meprin alpha by Western immunoblotting. In vitro studies demonstrated the ability of NE to activate meprin alpha, which in turn could release soluble TGFalpha and induce IL-8 production from 16HBE14o- cells. These effects were abrogated by actinonin, a specific meprin inhibitor. NE-induced IL-8 expression was also inhibited by meprin alpha siRNA. Immunoprecipitation studies detected EGFR/TLR4 complexes in NE-stimulated cells overexpressing these receptors. Confocal studies confirmed colocalization of EGFR and TLR4 in 16HBE14o- cells stimulated with meprin alpha. NFkappaB was also activated via MyD88 in these cells by meprin alpha. In bronchoalveolar lavage fluid from NE knock-out mice infected intra-tracheally with Pseudomonas aeruginosa meprin alpha was significantly decreased compared with control mice, and was significantly increased and correlated with NE activity, in bronchoalveolar lavage fluid from individuals with cystic fibrosis but not healthy controls. The data describe a previously unidentified lung metalloprotease meprin alpha, and its role in NE-induced EGFR and TLR4 activation and IL-8 production.

Fuelwood collection and consumption: a case study in Lupeta Tanzania

Fuelwood is a major resource in rural areas. Fuelwood collection and consumption habits were monitored in Lupeta, Tanzania through household interviews and fuelwood collection walks. Social dimensions, economic aspects of fuelwood, and alternative fuel sources were also examined. The study found that for all wealth classes, fuelwood is the primary source of fuel used within the village, with the middle and upper classes occasionally supplementing fuelwood with charcoal. Women collect and consume fuelwood for cooking. The majority of women (69%) prefer to use charcoal because fuelwood collection is labor intensive and time consuming. While the use of charcoal would provide more time for other required household activities, local economic constraints inhibit their adoption. The fuel shift from biomass fuels to transition fuels is happening slowly in Lupeta from fuelwood to charcoal. As fuelwood becomes scarcer, improved methods will need to be adopted or the fuel source will change.

Add a Dash of Rhetoric, a Pinch of Technical Communication, a Handful of Pedagogy, and Mix Well: Cookbooks and Recipes as Instructional Texts, and What Domestic Texts from 1870 to 1935 Can Teach Us About Technical Communication

Through the use of rhetoric centered on authority and risk avoidance, scientific method has co-opted knowledge, especially women's everyday and experiential knowledge in the domestic sphere. This, in turn, has produced a profound affect on technical communication in the present day. I am drawing on rhetorical theory to study cookbooks and recipes for their contributions to changes in instructional texts. Using the rhetorical lenses of metis (cunning intelligence), kairos (timing and fitness) and mneme (memory), I examine the way in which recipes and cookbooks are constructed, used and perceived. This helps me uncover lost voices in history, the voices of women who used recipes, produced cookbooks and changed the way instructions read. Beginning with the earliest cookbooks and recipes, but focusing on the pivotal temporal interval of 1870-1935, I investigate the writing and rhetorical forces shaping instruction sets and domestic discourse. By the time of scientific cooking and domestic science, everyday and experiential knowledge were being excluded to make room for scientific method and the industrial values of the public sphere. In this study, I also assess how the public sphere, via Cooperative Extension Services and other government agencies, impacted the domestic sphere, further devaluing everyday knowledge in favor of the public scientific model. I will show how the changes in the production of food, cookbooks and recipes were related to changes in technical communication. These changes had wide rippling effects on the field of technical communication. By returning to some of the tenets and traditions of everyday and experiential knowledge, technical communication scholars, practitioners and instructors today can find new ways to encounter technical communication, specifically regarding the creation of instructional texts. Bringing cookbooks, recipes and everyday knowledge into the classroom and the field engenders a new realm of epistemological possibilities.

Effects of epidermal growth factor on the invasion activity of the oral cancer cell lines HSC3 and SAS

Epidermal growth factor (EGF) is excreted in a high concentration in human saliva and modulates the growth and differentiation of various cancer cells. To elucidate the molecular mechanisms by which EGF affects oral cancer growth and invasion, we analyzed the Matrigel invasion activity of the cultured oral cancer cell line. Cells grown under the influence of EGF were subjected to Matrigel invasion assays and cells grown in the absence of EGF were used as controls. Gelatin-zymography and Northern blot analyses quantified the invasiveness and tumorigenicity. Chloramphenicol acetyltransferase assay (CAT assay) determined the EGF stimulation of matrix metalloproteinase (MMP) expression. EGF increased the number of cells penetrating a Matrigel membrane. Gelatin-zymography and Northern blot analysis revealed that MMP9 and Ets1 expressions correlated with EGF but MMP2 was not changed. a transient transfection assay revealed that EGF increased the promoter activities of the MMP9 genes in HSC3 and SAS cells. These results suggest that EGF increases the invasion activity of oral cancer cells partly by increasing MMP9.