Consumer perception of smoked Cheddar cheese.

Consumer perception of smoked cheese was evaluated through focus groups, surveys, and central location testing. Three focus groups (n = 29) were conducted with consumers of smoked cheese. Subsequently, 2 online surveys were conducted. The purpose of the first survey (n = 1,195) was to understand types of smoked cheeses consumed and if consumers associated specific wood smokes with smoked cheese. Next, an adaptive choice-based conjoint (n = 367) was designed to evaluate consumer perception of different attributes of smoked cheese. Maximum difference scaling and familiarity questions were also included in the adaptive choice-based conjoint survey. Following the surveys, a central location test (n = 135) was conducted with cheeses smoked with 3 different woods at a low and high intensity (6 cheeses total). Hierarchical Bayesian estimation, 1-way ANOVA, agglomerative hierarchical clustering, and 2-way ANOVA (smoke type × intensity level) were used to interpret the collected data. Results from the focus groups indicated that smoked cheese was perceived as an artisan, high-end product and that appearance and price were strong purchase factors. In general, consumers were not aware of how smoked flavor was imparted to cheese, but when informed of the processes, they preferred cold-smoked cheese to the addition of liquid smoke flavor. Results from both surveys confirmed focus group observations. Consumers perceived flavor differences among different wood smokes and smoked products. Method of smoking, smoke intensity, type of wood, and type of cheese were the most important attributes for purchase of smoked cheese. When tasting, consumers differentiated smoke aroma and flavor among cheeses and preferred cherry wood smoked cheeses over apple wood or hickory smoked cheeses. Understanding consumer perceptions of smoked cheese will give insight into the desired experience that consumers expect when purchasing smoked cheese.

Heritable gene expression differences between lake and stream stickleback include both parallel and antiparallel components.

The repeated phenotypic patterns that characterize populations undergoing parallel evolution provide support for a deterministic role of adaptation by natural selection. Determining the level of parallelism also at the genetic level is thus central to our understanding of how natural selection works. Many studies have looked for repeated genomic patterns in natural populations, but work on gene expression is less common. The studies that have examined gene expression have found some support for parallelism, but those studies almost always used samples collected from the wild that potentially confounds the effects of plasticity with heritable differences. Here we use two independent pairs of lake and stream threespine stickleback (Gasterosteus aculeatus) raised in common garden conditions to assess both parallel and antiparallel (that is, similar versus different directions of lake-stream expression divergence in the two watersheds) heritable gene expression differences as measured by total RNA sequencing. We find that more genes than expected by chance show either parallel (22 genes, 0.18% of expressed genes) or antiparallel (24 genes, 0.20% of expressed genes) lake-stream expression differences. These results correspond well with previous genomic studies in stickleback ecotype pairs that found similar levels of parallelism. We suggest that parallelism might be similarly constrained at the genomic and transcriptomic levels.

Computational Fluid Dynamics Studies of a Flow Reactor: Free Energies of Clusters of Sulfuric Acid with NH3 or Dimethyl Amine.

Computational fluid dynamics simulations of a flow reactor provided 3D spatial distributions of its temperature and flow profiles and abundances of sulfuric acid, nitrogeneous base, and the acid-base clusters formed from them. Clusters were simulated via their kinetic formation and decomposition involving sulfuric acid and base molecules. Temperature and flow profiles and the base and sulfuric acid distributions are characterized and the latter is compared to mass spectrometer measurements. Concentrations of simulated clusters of sulfuric acid with either NH3 or dimethylamine were compared to experimentally measured particle concentrations. Cluster thermodynamics were adjusted to better the agreement between simulated and experimental results. Free energies of acid-base clusters derived here are also compared to recent quantum chemistry calculations. Sensitivities to the thermodynamics were explored with a 2D laminar flow simulation and the abundance of large clusters was most sensitive to the thermodynamics of the smallest cluster, consisting of 1 base and 1 acid. Comparisons of this model to the computational fluid dynamics models provide verification of the implemented cluster chemistry. A box model was used to calculate nucleation rates for the conditions of other experimental work, and to provide predictions of nucleation for typical atmospheric conditions.

Cataloguing of Potential HIV Susceptibility Factors during the Menstrual Cycle of Pig-Tailed Macaques by Using a Systems Biology Approach.

UNLABELLED: Our earlier studies with pig-tailed macaques demonstrated various simian-human immunodeficiency virus (SHIV) susceptibilities during the menstrual cycle, likely caused by cyclic variations in immune responses in the female genital tract. There is concern that high-dose, long-lasting, injectable progestin-based contraception could mimic the high-progesterone luteal phase and predispose women to human immunodeficiency type 1 (HIV-1) acquisition and transmission. In this study, we adopted a systems biology approach employing proteomics (tandem mass spectrometry), transcriptomics (RNA microarray hybridization), and other specific protein assays (enzyme-linked immunosorbent assays and multiplex chemokine and cytokine measurements) to characterize the effects of hormonal changes on the expression of innate factors and secreted proteins in the macaque vagina. Several antiviral factors and pathways (including acute-phase response signaling and complement system) were overexpressed in the follicular phase. Conversely, during the luteal phase there were factors overexpressed (including moesins, syndecans, and integrins, among others) that could play direct or indirect roles in enhancing HIV-1 infection. Thus, our study showed that specific pathways and proteins or genes might work in tandem to regulate innate immunity, thus fostering further investigation and future design of approaches to help counter HIV-1 acquisition in the female genital tract. IMPORTANCE: HIV infection in women is poorly understood. High levels of the hormone progesterone may make women more vulnerable to infection. This could be the case during the menstrual cycle, when using hormone-based birth control, or during pregnancy. The biological basis for increased HIV vulnerability is not known. We used an animal model with high risk for infection during periods of high progesterone. Genital secretions and tissues during the menstrual cycle were studied. Our goal was to identify biological factors upregulated at high progesterone levels, and we indeed show an upregulation of genes and proteins which enhance the ability of HIV to infect when progesterone is high. In contrast, during low-progesterone periods, we found more HIV inhibitory factors. This study contributes to our understanding of mechanisms that may regulate HIV infection in females under hormonal influences. Such knowledge is needed for the development of novel prevention strategies.

Testing for parallel allochronic isolation in lake-stream stickleback.

The evolution of reproductive isolation (RI) is a critical step shaping progress towards speciation. In the context of ecological speciation, a critical question is the extent to which specific reproductive barriers important to RI evolve rapidly and predictably in response to environmental differences. Only reproductive barriers with these properties (importance, rapidity, predictability) will drive the diversification of species that are cohesively structured by environment type. One candidate barrier that might exhibit such properties is allochrony, whereby populations breed at different times. We studied six independent lake-stream population pairs of threespine stickleback (Gasterosteus aculeatus Linnaeus, 1758) that are known from genetic studies to show RI. However, the specific reproductive barriers driving this RI have proven elusive, leading to a 'conundrum of missing reproductive isolation'. We here show that breeding times differ among some of the populations, but not in a consistent manner between lakes and streams. Moreover, the timing differences between lake and stream populations within each pair could account for only a small proportion of total RI measured with neutral genetic markers. Allochrony cannot solve the conundrum of missing reproductive isolation in lake-stream stickleback.

Assessing reproductive isolation using a contact zone between parapatric lake-stream stickleback ecotypes.

Ecological speciation occurs when populations evolve reproductive isolation as a result of divergent natural selection. This isolation can be influenced by many potential reproductive barriers, including selection against hybrids, selection against migrants and assortative mating. How and when these barriers act and interact in nature is understood for relatively few empirical systems. We used a mark-recapture experiment in a contact zone between lake and stream three-spined sticklebacks (Gasterosteus aculeatus, Linnaeus) to evaluate the occurrence of hybrids (allowing inferences about mating isolation), the interannual survival of hybrids (allowing inferences about selection against hybrids) and the shift in lake-like vs. stream-like characteristics (allowing inferences about selection against migrants). Genetic and morphological data suggest the occurrence of hybrids and no selection against hybrids in general, a result contradictory to a number of other studies of sticklebacks. However, we did find selection against more lake-like individuals, suggesting a barrier to gene flow from the lake into the stream. Combined with previous work on this system, our results suggest that multiple (most weakly and often asymmetric) barriers must be combining to yield substantial restrictions on gene flow. This work provides evidence of a reproductive barrier in lake-stream sticklebacks and highlights the value of assessing multiple reproductive barriers in natural contexts.

Evidence supporting vertical transmission of Salmonella in dairy cattle.

We set out to investigate whether Salmonella enterica could be recovered from various tissues of viable neonatal calves immediately following parturition. Eleven samples were aseptically collected from each of 20 calves and consisted of both left and right subiliac and prescapular lymph nodes (LN), mesenteric LN, spleen and liver, as well as intestinal tissue (including luminal contents) from the small intestine, caecum, spiral colon and rectum. In addition, a faecal sample was collected from 19 of the dams. Salmonella was recovered from at least one sample from 10 of the 20 neonates. Across all calves, Salmonella was recovered from 12·7% of all samples and from LN in particular, Salmonella was recovered from 10·0%, 5·0%, and 5·0% of subiliac, prescapular, and mesenteric LN, respectively. Within calves, Salmonella was recovered from 0% to 73% of samples and across tissues, estimates of Salmonella prevalence were greatest in the caecum (30%) but was never recovered from the right pre-scapular LN. These data provide evidence of vertical transmission from a dam to her fetus such that viable calves are born already infected and thereby not requiring faecal-oral exposure for transmission. This new knowledge ought to challenge - or at least add to - existing paradigms of Salmonella transmission dynamics within cattle herds.

Plane of nutrition influences the performance, innate leukocyte responses, and resistance to an oral Salmonella enterica serotype Typhimurium challenge in Jersey calves.

Two experiments investigated how plane of nutrition influences performance, leukocyte responses, and resistance to an oral Salmonella enterica serotype Typhimurium challenge. In experiment 1, 46 (2±1 d of age) calves were randomly assigned to 2 diets: a low (LPN; n=23) and high plane of nutrition (HPN; n=23). The LPN calves were fed 409 g/d of dry matter (DM) of a 20% crude protein and 20% fat milk replacer, whereas HPN calves were fed 610 and 735 g/d of DM of a 28% crude protein and 25% fat milk replacer during wk 1 and 2 to 6, respectively. In experiment 2, 20 bull calves (LPN; n=11 and HPN; n=9) were orally challenged on d 80 with 1.5×10(7) cfu of Salmonella Typhimurium (ATCC #14028). The HPN calves had a greater incidence (87.5 vs. 45.5%) and duration of days with high fecal scores (5.5 vs. 3.5 d). The LPN calves had greater neutrophil surface expression of L-selectin on d 7, 21, and 42. Following the Salmonella Typhimurium challenge, calf starter DM intake was greater among the HPN calves. The percentage of neutrophils producing an oxidative burst was also greater among HPN calves on d 1 to 5 after the challenge. Similarly, the intensity of the oxidative burst tended to be greater among the HPN calves on d 2 and 3 postchallenge. The secretion of tumor necrosis factor-α from whole-blood cultures stimulated with lipopolysaccharide tended to be greater on d 1 and was greater on d 5 and 6 among HPN calves. The median ranks of haptoglobin concentrations were greater and plasma zinc concentrations tended to be decreased among LPN calves. These data indicate that feeding a HPN to Jersey calves improved average daily gain and feed efficiency, but increased the incidence of high fecal scores during the first few weeks of life; however, the HPN Jersey calves may be more resistant to Salmonella Typhimurium after weaning.

Insights into the pathophysiology of catch-up compared with non-catch-up growth in children born small for gestational age: an integrated analysis of metabolic and transcriptomic data.

Small for gestational age (SGA) children exhibiting catch-up (CU) growth have a greater risk of cardiometabolic diseases in later life compared with non-catch-up (NCU) SGA children. The aim of this study was to establish differences in metabolism and gene expression profiles between CU and NCU at age 4-9 years. CU children (n=22) had greater height, weight and body mass index standard deviation scores along with insulin-like growth factor-I (IGF-I) and fasting glucose levels but lower adiponectin values than NCU children (n=11; all P<0.05). Metabolic profiling demonstrated a fourfold decrease of urine myo-inositol in CU compared with NCU (P<0.05). There were 1558 genes differentially expressed in peripheral blood mononuclear cells between the groups (P<0.05). Integrated analysis of data identified myo-inositol related to gene clusters associated with an increase in insulin, growth factor and IGF-I signalling in CU children (P<0.05). Metabolic and transcriptomic profiles in CU SGA children showed changes that may relate to cardiometabolic risk.

Improved performance and heightened neutrophil responses during the neonatal and weaning periods among outdoor group-housed Holstein calves.

The objective was to determine if outdoor group housing of Holstein calves influences metabolic status, leukocyte responses, and behavior compared with individually housed calves. Forty-nine Holstein heifer calves (2 ± 1 d of age) were randomly assigned to 1 of 2 treatments: individually housed (G1; n = 22) or group housed [3 calves per pen (G3); n = 27]. The space allowances per calf were 4.8 and 7.0m(2) for G1 and G3, respectively. All calves were offered an identical plane of milk replacer nutrition (747 and 1,010 g of DM/d of a 28% CP:20% fat milk replacer from wk 1 to 2 and wk 3 to 6, respectively). Weaning was initiated during wk 7 by removing the p.m. feeding and calves were completely weaned when they consumed 900 g of calf starter/d (as fed) for 2 consecutive days after d 54. At d 90, calves were commingled into random outdoor groups of 5 calves per pen. Peripheral blood was collected during the neonatal (d 3, 10, and 21), weaning (d 46, 48, and 54), and commingling periods (d 90, 93, and 98) and was analyzed for neutrophil oxidative burst (OB) capacity when cocultured with Escherichia coli, neutrophil surface L-selectin protein expression, and whole-blood secretion of tumor necrosis factor-α when cocultured with lipopolysaccharide. Starter intake was greater for G3 during the postweaning period (wk 8 to 12). Average daily gain was greater for G3 than G1 from d 54 to 68 and tended to be greater after commingling from d 113 to 133. During the neonatal period, G3 calves had more activated neutrophils, as evidenced by increased neutrophil L-selectin protein expression and a tendency for increased percentage of neutrophils producing an OB than G1 calves. During weaning, G3 calves continued to have more activated neutrophils with increased L-selectin expression on d 46 and 48 and a greater OB intensity throughout the period. No differences were observed among leukocyte responses between treatments at d 93 and 98. Outdoor group-housed Holstein calves had improved performance and heightened neutrophil responses compared with individually housed calves.

Detection of B-mode polarization in the cosmic microwave background with data from the South Pole Telescope.

Gravitational lensing of the cosmic microwave background generates a curl pattern in the observed polarization. This "B-mode" signal provides a measure of the projected mass distribution over the entire observable Universe and also acts as a contaminant for the measurement of primordial gravity-wave signals. In this Letter we present the first detection of gravitational lensing B modes, using first-season data from the polarization-sensitive receiver on the South Pole Telescope (SPTpol). We construct a template for the lensing B-mode signal by combining E-mode polarization measured by SPTpol with estimates of the lensing potential from a Herschel-SPIRE map of the cosmic infrared background. We compare this template to the B modes measured directly by SPTpol, finding a nonzero correlation at 7.7σ significance. The correlation has an amplitude and scale dependence consistent with theoretical expectations, is robust with respect to analysis choices, and constitutes the first measurement of a powerful cosmological observable.

Comparison of pork quality and sensory characteristics for antibiotic free yorkshire crossbreds raised in hoop houses.

The objective of this study was to compare pork characteristics and to determine consumer acceptability of pork chops from antibiotic free Yorkshire crossbreds sired by Berkshire (BY), Large Black (LBY), Tamworth (TY) or Yorkshire (YY) boars and reared in hoop houses. The experiments were conducted at the North Carolina Agricultural and Technical State University (NCA&TSU) Farm in Greensboro, NC and the Cherry Research Station Center for Environmental Farming Systems (CEFS) Alternative Swine Unit in Goldsboro, NC (source of antibiotic free Yorkshire sows used at both places). Twenty-four sows were artificially inseminated at each location in each of three trials. Litters were weaned at 4 wks old, and reared within deep-bedded outdoor hoop houses. To compare pork characteristics, 104 randomly selected animals were harvested at a USDA-inspected abattoir at approximately 200 d of age. Variables measured included pH, color score, L*, a*, b*, marbling score, drip loss, hot carcass weight, backfat thickness (BF), loin muscle area (LMA), and slice shear force. Sensory panel tests were also conducted at two time periods. The data was analyzed with GLM in SAS 9.01 including location, trial, and sire breed as fixed effects. Backfat thickness, LMA, color score and a* were different among breeding groups (p<0.05). The LBY pigs had thicker backfat and smaller LMA than the other breed types. The TY and YY had less backfat than all other breed groups. Color score was lower for YY than BY and LBY but intermediate for TY. The a* was lower for TY than other breeds except LBY which was intermediate. For one sensory panel test, YY pork was more preferred overall as well as for juiciness and texture compared to BY and LBY (p<0.05), but no impact of breed type was noted for the other test, with values similar for BY, LBY, TY and YY pork. This information may help small farmers make decisions about breed types to use for outdoor production.

Ambient pressure proton transfer mass spectrometry: detection of amines and ammonia.

An instrument to detect gaseous amines and ammonia is described, and representative data from an urban site and a laboratory setting are presented. The instrument, an Ambient pressure Proton transfer Mass Spectrometer (AmPMS), consists of a chemical ionization and drift region at atmospheric pressure coupled to a standard quadrupole mass spectrometer. Calibrations show that AmPMS sensitivity is good for amines, and AmPMS backgrounds were suitably determined by diverting sampled air through a catalytic converter. In urban air at a site in Atlanta, amines were detected at subpptv levels for methyl and dimethyl amine which were generally at a low abundance of <1 and ∼3 pptv, respectively. Trimethyl amine (or isomers) was on average about 4 pptv in the morning and increased to 15 pptv in the afternoon, while triethyl amine (or isomers or amides) increased to 25 pptv on average in the late afternoon. The background levels for the 4 and 5 carbon amines and ammonia were high, and data are very limited for these species. Improvements in detecting amines and ammonia from a smog chamber were evident due to improvements in AmPMS background determination; notably dimethyl amine and its OH oxidation products were followed along with impurity ammonia and other species. Future work will focus on accurate calibration standards and on improving the sample gas inlet.

Failure of rabbit neutrophils to secrete endogenous pyrogen when stimulated with staphylococci.

Cells obtained from acute peritoneal exudates in rabbits were separated into neutrophil and mononuclear populations by centrifugation on colloidal silica gradients. When these populations were separately incubated in tissue culture medium in the presence of opsonized Staphylococcus epidermidis, endogenous pyrogen was secreted only by the adherent cells of the mononuclear population. Pyrogen production by neutrophils could not have amounted to as much as 1% of the pyrogen produced by macrophages. When mononuclear cells were added back to purified neutrophils, no pyrogen was produced that could not be accounted for by the number of macrophages added. Rabbit blood cells were similarly fractionated on colloidal silica gradients. Again, endogenous pyrogen was made only by the adherent mononuclear population. The neutrophils isolated on these gradients appeared to be morphologically normal and were 85% viable as judged by dye exclusion. They showed normal random motility. Both blood and exudate neutrophils responded chemotactically to N-formyl Met-Leu-Phe, and blood neutrophils responded chemotactically to zymosan-activated serum. Both kinds of neutrophils phagocytosed zymosan particles and both killed opsonized S. epidermidis in a roller tube system. Both blood and exudate neutrophils showed normal superoxide production when stimulated with opsonized zymosan particles. This evidence suggests that macrophages are the only source of endogenous pyrogens, and that pyrogens secreted by cell populations that are rich in neutrophils are to be attributed to the monocytes or macrophages that they contain.

Self-recognition of CD1 by gamma/delta T cells: implications for innate immunity.

The specificity of immunoglobulins and alpha/beta T cell receptors (TCRs) provides a framework for the molecular basis of antigen recognition. Yet, evolution has preserved a separate lineage of gamma/delta antigen receptors that share characteristics of both immunoglobulins and alpha/beta TCRs but whose antigens remain poorly understood. We now show that T cells of the major tissue gamma/delta T cell subset recognize nonpolymorphic CD1c molecules. These T cells proliferated in response to CD1+ presenter cells, lysed CD1c+ targets, and released T helper type 1 (Th1) cytokines. The CD1c-reactive gamma/delta T cells were cytotoxic and used both perforin- and Fas-mediated cytotoxicity. Moreover, they produced granulysin, an important antimicrobial protein. Recognition of CD1c was TCR mediated, as recognition was transferred by transfection of the gamma/delta TCR. Importantly, all CD1c-reactive gamma/delta T cells express V delta 1 TCRs, the TCR expressed by most tissue gamma/delta T cells. Recognition by this tissue pool of gamma/delta T cells provides the human immune system with the capacity to respond rapidly to nonpolymorphic molecules on professional antigen presenting cells (APCs) in the absence of foreign antigens that may activate or eliminate the APCs. The presence of bactericidal granulysin suggests these cells may directly mediate host defense even before foreign antigen-specific T cells have differentiated.

Treatment of whole blood (WB) and red blood cells (RBC) with S-303 inactivates pathogens and retains in vitro quality of stored RBC.

A pathogen inactivation (PI) process has been developed using the frangible anchor linker effector (FRALE) compound S-303. A series of experiments were performed in whole blood (WB) to measure the level of viral and bacterial inactivation. The results showed that 0.2mM S-303 and 2mM glutathione (GSH) inactivated >6.5 logs of HIV, >5.7 logs of Bluetongue virus, >7.0 logs of Yersinia enterocolitica, 4.2 logs of Serratia marcescens, and 7.5 logs of Staphylococcus epidermidis. Recent development for S-303 is focused on optimization of the PI process for red blood cell concentrates (RBC). A series of studies in RBC showed that 0.2mM S-303 and 20mM GSH inactivated approximately 5 logs or greater of Y. enterocolitica, E. coli, S. marcescens, S. aureus, HIV, bovine viral diarrhoea virus, bluetongue virus and human adenovirus 5. In both applications of the S-303 process, in vitro parameters of RBC function and physiology were retained compared to conventional RBC. Results from these studies indicate that S-303 can be applicable for PI of RBC and WB.

The Reaction of CIONO2 with Submicrometer Sulfuric Acid Aerosol.

The measured reaction probability, gamma for the reaction of chlorine nitrate (CIONa(2)) with 60 percent (by weight) sulfuric acid aerosol increases monotonically with particle size at 250 kelvin. The reacto-diffusive length (l, the effective liquid depth over which reaction occurs) derived from these experiments is 0.037 +/- 0.007 micrometer (95 percent confidence level for precision). The reaction probability for the reaction of CIONO(2) with 60 percent sulfuric acid aerosol doped with approximately 7 x 10(-4) M hydrochloric acid at 250 kelvin is larger by about a factor of 4 than in the absence of hydrochloric acid and varies less with particle size (l = 0.009 +/- 0.005 micrometer). These results provide a test of the theory for gas-particle reactions and further insight into the reactivity of atmospheric aerosol.

An antimicrobial activity of cytolytic T cells mediated by granulysin.

Cytolytic T lymphocytes (CTLs) kill intracellular pathogens by a granule-dependent mechanism. Granulysin, a protein found in granules of CTLs, reduced the viability of a broad spectrum of pathogenic bacteria, fungi, and parasites in vitro. Granulysin directly killed extracellular Mycobacterium tuberculosis, altering the membrane integrity of the bacillus, and, in combination with perforin, decreased the viability of intracellular M. tuberculosis. The ability of CTLs to kill intracellular M. tuberculosis was dependent on the presence of granulysin in cytotoxic granules, defining a mechanism by which T cells directly contribute to immunity against intracellular pathogens.

Impact of storage temperature and product pH on the survival of Listeria monocytogenes in vacuum-packaged souse.

Souse is a fully cooked, ready-to-eat gelled pork product. There is a zero-tolerance policy for Listeria monocytogenes in ready-to-eat meat products. The survival and/or growth of L. monocytogenes in souse is unknown. The effectiveness of three different souse formulations (pH 4.3, 4.7, and 5.1) for controlling the growth of L. monocytogenes at two refrigerated storage temperatures (5 and 10 degrees C) was evaluated. All products were vacuum packaged. Uninoculated product was prepared as the control, and other products were artificially surface contaminated with a three-strain cocktail of L. monocytogenes (10(6) CFU/ cm2). Microbial counts were obtained on selective and nonselective media twice weekly through 8 weeks of storage. Souse did not support the growth of L. monocytogenes regardless of product formulation or storage temperature. At 5 degrees C, D-values for products with pH values of 4.7 and 5.1 were not different, but survival of L. monocytogenes in product with a lower pH (4.3) was decreased compared with survival in products with higher pH values (P < 0.05). Survival of L. monocytogenes was not impacted by storage temperatures (P > 0.05). Consumer acceptability (n = 75 souse consumers) of pH 4.3 products was not different from that for (typical) pH 4.7 products (P > 0.05). These results indicate that conventionally produced souse does not support the growth of L. monocytogenes and that inactivation of the organism is more likely in products formulated at a lower pH (< or = 4.3) without affecting consumer acceptance.

Topology of the pore-region of a K+ channel revealed by the NMR-derived structures of scorpion toxins.

The architecture of the pore-region of a voltage-gated K+ channel, Kv1.3, was probed using four high affinity scorpion toxins as molecular calipers. We established the structural relatedness of these toxins by solving the structures of kaliotoxin and margatoxin and comparing them with the published structure of charybdotoxin; a homology model of noxiustoxin was then developed. Complementary mutagenesis of Kv1.3 and these toxins, combined with electrostatic compliance and thermodynamic mutant cycle analyses, allowed us to identify multiple toxin-channel interactions. Our analyses reveal the existence of a shallow vestibule at the external entrance to the pore. This vestibule is approximately 28-32 A wide at its outer margin, approximately 28-34 A wide at its base, and approximately 4-8 A deep. The pore is 9-14 A wide at its external entrance and tapers to a width of 4-5 A at a depth of approximately 5-7 A from the vestibule. This structural information should directly aid in developing topological models of the pores of related ion channels and facilitate therapeutic drug design.