ANAVIP® interaction with western pygmy rattlesnake venom: In vitro assesment of reactivity using SE-HPLC.

Every year large numbers of venomous snake bites occur around the world, especially in tropical areas. The World Health Organization classifies venomous snake bites as one of their highest priority neglected tropical diseases, one of the reasons being the short supply of antivenom compared to the number of snake envenomations. The standard of care for snake envenomation is administration of antivenom. Many antivenoms are polyvalent, which are produced using venoms from multiple species of snakes. The polyvalent antivenoms can treat envenomation from snake venoms used in the production, but also show cross-reactivity against snake venoms with similar composition. Determining cross-reactivities of antivenoms could help improve the quality of treatment and provide a better understanding of venom-antivenom binding. One antivenom only has been available in the United States for treatment of North American Crotaline envenomation, with the recent introduction of an F(ab')2 antivenom (ANAVIP®). Size-exclusion high performance liquid chromatography (SE-HPLC) was used to assess cross-reactivity of the western pygmy rattlesnake, Sistrurus miliarius streckeri (S. m. streckeri), against ANAVIP®. Estimates of venom-antivenom reactivity was measured in reaction mixtures based on the increase in elution profile area of higher molecular weight complexes (region 1) and on the decrease in elution profile area of reactants (region 2). Reaction mixtures contained ANAVIP® (1.0 mg/ml) and S. m. streckeri venom (0.125, 0.25, 0.5, or 1.0 mg/ml). Controls were ANAVIP® and S. m. streckeri (1.0 mg/ml). Mixtures were incubated at 37 °C for 30 min, then stored at 4 °C (5 min) prior to SE-HPLC. Relative binding, estimated from the increase in region 1 (immune complexes) and decrease in region 2 (reactants) region areas, suggested saturation of reactive antivenom binding sites at 0.125 (and above) mg venom/mg antivenom. SE-HPLC data indicate that binding of ANAVIP® to S. m. streckeri venom does occur, consistent with protective effects observed clinically. Further studies are needed to compare the binding of S. m. streckeri venom to other commercially available antivenoms, and the binding of ANAVIP® to other venoms of clinical significance.

A bioactive metallurgical grade porous silicon-polytetrafluoroethylene sheet for guided bone regeneration applications.

The properties of porous silicon make it a promising material for a host of applications including drug delivery, molecular and cell-based biosensing, and tissue engineering. Porous silicon has previously shown its potential for the controlled release of pharmacological agents and in assisting bone healing. Hydroxyapatite, the principle constituent of bone, allows osteointegration in vivo, due to its chemical and physical similarities to bone. Synthetic hydroxyapatite is currently applied as a surface coating to medical devices and prosthetics, encouraging bone in-growth at their surface and improving osseointegration. This paper examines the potential for the use of an economically produced porous silicon particulate-polytetrafluoroethylene sheet for use as a guided bone regeneration device in periodontal and orthopaedic applications. The particulate sheet is comprised of a series of microparticles in a polytetrafluoroethylene matrix and is shown to produce a stable hydroxyapatite on its surface under simulated physiological conditions. The microstructure of the material is examined both before and after simulated body fluid experiments for a period of 1, 7, 14 and 30 days using Scanning Electron Microscopy. The composition is examined using a combination of Energy Dispersive X-ray Spectroscopy, Thin film X-ray diffraction, Attenuated Total Reflectance-Fourier Transform Infrared Spectroscopy and the uptake/release of constituents at the fluid-solid interface is explored using Inductively Coupled Plasma-Optical Emission Spectroscopy. Microstructural and compositional analysis reveals progressive growth of crystalline, 'bone-like' apatite on the surface of the material, indicating the likelihood of close bony apposition in vivo.

Defocus image contrast in hexagonally-ordered mesoporous material.

A transmission electron microscope was used to characterize a powder form of hexagonally-ordered mesoporous silica material. The structural symmetry built into this amorphous material allowed one to obtain three characteristic images, i.e. a hexagonal honey-comb structure and wide/narrow parallel lines. These images were found to originate primarily from phase contrast, which changed sensitively with defocusing. To further understand the contrast behaviour of these images, an analytical form of the defocus contrast theory was developed and applied to the simulation of the characteristic wide/narrow parallel line images. The simulation was found to be in good qualitative agreement with experiments, where changes in focus conditions and specimen thickness were predicted to alter the contrast in the resulting parallel-line type images.

Quantitative TEM analysis of a hexagonal mesoporous silicate structure.

TEM analysis of mesoporous materials is generally undertaken to give qualitative results. Accurate quantitative analysis is demonstrated in this study. A systematic image analysis of a powder form of a hexagonal mesoporous material known as KIT-6 is conducted using a transmission electron microscope (TEM). Three types of image contrast typically appear in this material (a hexagonal honeycomb structure, wide and narrow parallel lines). The honeycomb face is used to characterise this material in terms of a conventional 2-D hexagonal structure and the d-spacings for the (100) and (110) planes are experimentally measured in varying focus conditions. A tilting experiment is conducted to determine how the angle of tilt affects the line spacing and their visibility. Tilting has very little effect on the line spacing, whereas it affects the visibility of both the wide and narrow lines by limiting an angle range of visibility. The hexagonal lattice structure parameter determined by TEM method is found to be approximately 7% lower than that calculated by low-angle X-ray diffraction. Thus we conclude that TEM data can be used to determine the geometry and dimensions of hexagonal mesoporous silica materials, with a small error in the hexagonal lattice parameter.

The antibacterial effects of zinc ion migration from zinc-based glass polyalkenoate cements.

Zinc-based glass polyalkenoate cements have been synthesised and their potential use in orthopaedic applications investigated. Zinc ions were released from the materials in a rapid burst over the first 24 h after synthesis, with the release rate falling below detectable levels after 7 days. Cement-implanted bone samples were prepared and the released zinc was shown, using energy dispersive X-ray analysis, to penetrate from the cement into the adjacent bone by up to 40 microm. Finally, the cements exhibited antibacterial activity against Streptococcus mutans and Actinomyces viscosus that reflected the pattern of zinc release, with the inhibition of growth greatest shortly after cement synthesis and little or no inhibition measureable after 30 days.

Ultrasonically set glass polyalkenoate cements for orthodontic applications.

There is an accepted clinical requirement for a luting cement that can be command set upon satisfactory placement of an orthodontic appliance onto dentition. This work evaluates the suitability of ultrasound, imparted from a dental scaler, as a potential mechanism for achieving this. The net setting times and subsequent compressive strengths of a range of commercial and experimental glass polyalkenoate cements (GPCs) were evaluated, using modified ISO 9917 methods, when set both chemically and by ultrasound. The ultrasound was applied to the GPC through an orthodontic brace. It was possible to command set GPCs by the application of five to ten seconds of ultrasound; the exact time required being dependent upon the composition of the GPC in question. The compressive strengths of these cements can be improved by around 90% with the command set when the optimum PAA molecular weight and tartaric acid content is employed.

Clothing fabric does not affect thermoregulation during exercise in moderate heat.

PURPOSE: We investigated whether temperature regulation is improved during exercise in moderate heat by the use of clothing constructed from fabric that was purported to promote sweat evaporation compared with traditional fabrics. METHODS: Eight well-trained, euhydrated males performed three exercise bouts wearing garments made from an evaporative polyester fabric (SYN), wearing garments made from traditional cotton fabric (COT), or dressed seminude (S-N) in random order. Bouts consisted of 15 min seated rest, 30 min running at 70% .VO(2max), 15 min walking at 40% .VO(2max), and 15 min seated rest, all at 30 +/- 1 degrees C and 35 +/- 5% relative humidity. COT and SYN clothing ensembles consisted of crew neck, short sleeve T-shirts, cycling shorts, and anklet socks made from their respective materials, and running shoes. The S-N condition consisted of a Lycra swim suit, polyester socks, and running shoes. RESULTS: Mean skin temperature was lower for S-N during preexercise rest when compared with SYN and COT. No differences in mean body temperature, rectal temperature, or mean skin temperature were observed during or after exercise. No differences in VO2 or heart rate were observed. No differences in comfort sensations were observed. CONCLUSION: In summary, before, during, or after exercise in a moderately warm environmental condition, neither the addition of a modest amount of clothing nor the fabric characteristics of this clothing alters physiological, thermoregulatory, or comfort sensation responses.

Expiratory flow limitation confounds ventilatory response during exercise in athletes.

INTRODUCTION: A significant number of highly trained endurance runners have been observed to display an inadequate hyperventilatory response to intense exercise. Two potential mechanisms include low ventilatory responsiveness to hypoxia and ventilatory limitation as a result of maximum expiratory flow rates being achieved. PURPOSE: To test the hypothesis that expiratory flow limitation can complicate determination of ventilatory responsiveness during exercise the following study was performed. METHODS/MATERIALS: Sixteen elite male runners were categorized based on expiratory flow limitation observed in flow volume loops collected during the final minute of progressive exercise to exhaustion. Eight flow limited (FL) (VO2max, 75.9+/-2.4 mL x kg(-1) x min(-1); expiratory flow limitation, 47.3+/-20.4%) and eight non-flow limited subjects (NFL) (VO2max, 75.6+/-4.8 mL x kg(-1) x min(-1); expiratory flow limitation, 0.3+/-0.8%) were tested for hypoxic ventilatory responsiveness (HVR). RESULTS: Independent groups ANOVA revealed no significant differences between FL and NFL for VO2max, VE max (136.2+/-16.0 vs 137.5+/-21.6 L x min(-1)), VE/VO2, (28.4+/-3.2 vs 27.6+/-2.9 L x lO2(-1)), VE/VCO2 (24.8+/-3.1 vs 24.4+/-2.0 L x lCO2(-1)), HVR (0.2+/-0.2 vs 0.3+/-0.1 L x %SaO2(-1)), or SaO2 at max (89.1+/-2.4 vs 86.6+/-4.1%). A significant relationship was observed between HVR and SaO2 (r = 0.92, P < or = 0.001) in NFL that was not present in FL. Conversely, a significant relationship between VE/VO2 and SaO2 (r = 0.79, P < or = 0.019) was observed in FL but not NFL. Regression analysis indicated that the HVR-SaO2 and SaO2-VE/VO2 relationships differed between groups. DISCUSSION: When flow limitation is controlled for, HVR plays a more significant role in determining SaO2 in highly trained athletes than has been previously suggested.

Partitioned weight loss and body composition changes during a mountaineering expedition: a field study.

Weight loss and changes in body composition are recognized phenomena associated with high-altitude mountaineering expeditions. Attempts to partition the weight loss between fat mass (FM) and fat-free mass (FFM) have been inconclusive. Therefore, five male subjects, average age 40.0 +/- 5.5 years, were studied prior to, during, and following a 21-day expedition between 2200 m and 4300 m on Mt. McKinley, Alaska. Pre- to postexpedition body composition changes were determined by densitometry, skinfold thickness (12 sites), body girth measurements (14 sites), and cross-sectional area from magnetic resonance imaging (MRI) (three sites). Data analysis by Student's t-test (p < 0.05(1) tail) indicated decreases in body weight (4.2 +/- 2.8 kg, 220 g/day, 5.4% of initial weight), total skinfold thickness (10.8%), total body girth (2.8%), and percentage of fat by densitometry (15%). Total cross-sectional area of the three MRI slices (upper arm + thigh + calf) decreased 4.7%, muscle area decreased 9.1%, and fat area decreased 3.9%. The three methods used to partition the weight loss between FM and FFM did not agree: 77% FM vs 23% FFM by densitometry, 25% FM vs 75% FFM by skinfolds, and 38% FM vs 62% FFM by MRI. Energy intake (3640 +/- 1250 kcal/day) was negatively correlated (Pearson r > 0.88(2) tail) with losses in weight (r = -0.89), skinfolds (r = -0.93), and girths (r = -0.88), ie the greater the intake the less the decline. Therefore, insufficient energy intake appeared primarily responsible for the weight loss and changes in body composition during the expedition.

Comparative Study of Micro-ID, Minitek and Conventional Methods with Enterobacteriaceae Freshly Isolated from Foods.

A total of 400 Enterobacteriaceae isolates freshly taken from broiler carcasses, ground beef, pork sausage, raw shrimp, pre-wrapped sandwiches, raw carrots, lettuce and fresh strawberries was inoculated into the 15 biochemical tests of the Micro-ID (4-h) system and into the 15 corresponding tests in the Minitek (24-h) and conventional systems. For each food there were 750 biochemical test comparisons (50 isolates × 15 tests). The overall agreement between Micro-ID and conventional tests was 96.8%, whereas the agreement between Minitek and conventional tests was 93.6%. Three laboratory technicians who independently recorded results of 6000 biochemical tests from each of the three systems were in complete agreement for 99.3%, 98.9% and 99.7% of the Micro-ID. Minitek and conventional tests, respectively. Thus results obtained with the miniaturized systems were as easy to read and interpret as conventional tests in tubes. The most frequently encountered Enterobacteriaceae from these foods were Escherichia coli (broiler carcasses, pork sausage). Enterobacter agglomerans (carrots, lettuce, shrimp, strawberries), Enterobacter cloacae (pre-wrapped sandwiches), and Serratia liquefaciens (ground beef).

Effectiveness of Sampling Methods for Salmonella Detection on Processed Broilers.

A total of 240 processed broiler carcasses (water-chilled and unfrozen) were each sampled by three methods (whole-carcass rinse, neck-skin rinse, and macerated neck skin) for detection of Salmonella . In addition to this, various procedures were compared: destructive (incubating the entire carcass with the rinse fluid) versus non-destructive (incubating the rinse water with concentrated lactose or selenite cystine broth added after removal of the carcass) sampling and pre-enrichment versus no pre-enrichment during Salmonella detection procedures. There was no significant difference (p < 0.05) between the percentage of Salmonella -positive carcasses obtained by destructive sampling and the percentage obtained by non-destructive samples of whole carcasses. There was also no significant difference (p < 0.05) in results obtained by rinsing and blending excised neck-skin samples. There was highly significant difference (p = 0.001), however, between whole carcass and neck-skin analyses. With whole-carcass sampling, 45% of the carcasses were positive for the presence of Salmonella while with rinsing or blending the neck skin of these same carcasses, only 11% and 12%, respectively, were positive for the organism. Pre-enrichment of the whole carcass, of the whole-carcass rinse, or of the neck-skin samples did not result in significantly greater percentages of positive results than did direct enrichment of these samples.

Relation of Enterobacteriaceae Counts to Salmonella Contamination of Market Broilers.

We determined the Enterobacteriaceae counts and the Salmonella status (positive or negative) of 20 individual birds in each of 12 groups of broiler carcasses. The overall logarithmic mean Enterobacteriaceae count for the 240 carcasses was 2.7 with group means ranging from 1.8 to 3.6. One hundred and twenty three (51.2%) of the 240 carcasses were Salmonella -positive. The number of Salmonella - positive carcasses within groups ranged from 0 to 18. No relationship was found between Enterobacteriaceae counts and presence of Salmonella in broiler carcasses.