Murine bladder wall biomechanics following partial bladder obstruction

Evaluation of bladder wall mechanical behavior is important in understanding the functional changes that occur in response to pathologic processes such as partial bladder outlet obstruction (pBOO). In the murine model, the traditional approach of cystometry to describe bladder compliance can prove difficult secondary to small bladder capacity and surgical exposure of the bladder. Here, we explore an alternative technique to characterize murine mechanical properties by applying biaxial mechanical stretch to murine bladders that had undergone pBOO. 5–6 week old female C57/Bl6 mice were ovariectomized and subjected to pBOO via an open surgical urethral ligation and sacrificed after 4 weeks (n=12). Age matched controls (n=6) were also analyzed. Bladders were separated based on phenotype of fibrotic (n=6) or distended (n=6) at the time of harvest. Biaxial testing was performed in modified Kreb's solution at 37 °C. Tissue was preconditioned to 10 cycles and mechanical response was evaluated by comparing axial strain at 50 kPa. The normal murine bladders exhibited anisotropy and were stiffer in the longitudinal direction. All mice showed a loss of anisotropy after 4 weeks of pBOO. The two phenotypes observed after pBOO, fibrotic and distended, exhibited less and more extensibility, respectively. These proof-of-principle data demonstrate that pBOO creates quantifiable changes in the mechanics of the murine bladder that can be effectively quantified with biaxial testing.     

Assessment of winter wheat cultivars for changes in straw composition and digestibility in response to fungicide and growth regulator treatments

The effects of a combined fungicide and growth regulator treatment on the composition and dry matter digestibility of straw samples of winter wheat cultivars, Beaver, Brigadier, Hussar, Mercia, Riband, Genesis, Hunter, Spark, Cadenza and Flame, were compared. The samples were analysed for nitrogen, ash, fibre fractions and minerals. Nitrogen content of treated samples was reduced compared to the untreated control samples and in contrast the proportion of ash increased for all the treated samples. All samples were analysed by derivative thermogravimetry (DTG) to determine changes in fibre composition. Only six cultivars were analysed by chemical fractionation methods to determine changes in lignin, cellulose and hemicellulose content. The results from the two methods of fibre analysis were broadly similar. K, Ca, Mg and Na concentrations of the treated straw samples were found to be different to the untreated samples. Increase in dry matter digestibility (DMD) of treated straw was observed for all cultivars; the effects on Beaver, Brigadier, Mercia, Riband, Genesis, Hunter and Cadenza were most pronounced, but were less for Hussar, Spark and Flame. The potential impact of these changes in straw quality on organic mushroom compost preparation is briefly discussed.     

USE of an ELECTRONIC HGMF INTERPRETER SYSTEM FOR ENUMERATION of NALIDIXIC ACID RESISTANT SALMONELLAE IN CHICKEN CAECA

An electronic counting system using hydrophobic grid-membrane filters (HGMF) and the HGMF Interpreter was evaluated for its usefulness in enumerating nalidixic acid resistant Salmonella in frozen chicken caeca. Salmonella recovery was equivalent on both Hektoen Enteric and EF-18 agars. However, the color of the Salmonella growth on EF-18 agar was more easily differentiated by the HGMF Interpreter electronic counting system. the study showed that a 4 h resuscitation on a nonselective medium was required in order to maximize the subsequent recovery on Hektoen Enteric agar, though not on EF-18 agar. Using the EF-18 agar as the Salmonella selective medium, a method was established that recorded counts of nalidixic acid resistant Salmonella rapidly and easily in electronic data files, for subsequent retrieval and manipulation.     

Microtubule Stabilization in Pressure Overload Cardiac Hypertrophy

Increased microtubule density, for which microtubule stabilization is one potential mechanism, causes contractile dysfunction in cardiac hypertrophy. After microtubule assembly, α-tubulin undergoes two, likely sequential, time-dependent posttranslational changes: reversible carboxy-terminal detyrosination (Tyr-tubulin ↔ Glu-tubulin) and then irreversible deglutamination (Glu-tubulin → Δ2-tubulin), such that Glu- and Δ2-tubulin are markers for long-lived, stable microtubules. Therefore, we generated antibodies for Tyr-, Glu-, and Δ2-tubulin and used them for staining of right and left ventricular cardiocytes from control cats and cats with right ventricular hypertrophy. Tyr- tubulin microtubule staining was equal in right and left ventricular cardiocytes of control cats, but Glu-tubulin and Δ2-tubulin staining were insignificant, i.e., the microtubules were labile. However, Glu- and Δ2-tubulin were conspicuous in microtubules of right ventricular cardiocytes from pressure overloaded cats, i.e., the microtubules were stable. This finding was confirmed in terms of increased microtubule drug and cold stability in the hypertrophied cells. In further studies, we found an increase in a microtubule binding protein, microtubule-associated protein 4, on both mRNA and protein levels in pressure-hypertrophied myocardium. Thus, microtubule stabilization, likely facilitated by binding of a microtubule-associated protein, may be a mechanism for the increased microtubule density characteristic of pressure overload cardiac hypertrophy.     

CONSUMER ACCEPTANCE OF COMMERCIALLY AVAILABLE FRANKFURTERS1

Consumers want meat products that are healthful and affordable. However, consumers will repurchase a product only if it tastes good. Consumers identified reasons for purchasing frankfurters and evaluated overall acceptability of 10 commercially available frankfurters. Consumers listed “taste” as the most important factor when purchasing frankfurters; only 16% of the consumers indicated that nutrition/health was the most important criterion. Nineteen consumer-perceived sensory and liking attributes were measured for the frankfurters in this study. The intensities of the attributes ranged from low to moderately high. All the frankfurters scored in the low to moderate range for overall acceptability. Most of the scores for liking of individual attributes correlated highly with overall liking, indicating that many of the questions relating to attribute liking could be eliminated.     

Spa47 is an oligomerization‐activated type three secretion system (T3SS) ATPase from Shigella flexneri

Gram‐negative pathogens often use conserved type three secretion systems (T3SS) for virulence. The Shigella type three secretion apparatus (T3SA) penetrates the host cell membrane and provides a unidirectional conduit for injection of effectors into host cells. The protein Spa47 localizes to the base of the apparatus and is speculated to be an ATPase that provides the energy for T3SA formation and secretion. Here, we developed an expression and purification protocol, producing active Spa47 and providing the first direct evidence that Spa47 is a bona fide ATPase. Additionally, size exclusion chromatography and analytical ultracentrifugation identified multiple oligomeric species of Spa47 with the largest greater than 8 fold more active for ATP hydrolysis than the monomer. An ATPase inactive Spa47 point mutant was then engineered by targeting a conserved Lysine within the predicted Walker A motif of Spa47. Interestingly, the mutant maintained a similar oligomerization pattern as active Spa47, but was unable to restore invasion phenotype when used to complement a spa47 null S. flexneri strain. Together, these results identify Spa47 as a Shigella T3SS ATPase and suggest that its activity is linked to oligomerization, perhaps as a regulatory mechanism as seen in some related pathogens. Additionally, Spa47 catalyzed ATP hydrolysis appears to be essential for host cell invasion, providing a strong platform for additional studies dissecting its role in virulence and providing an attractive target for anti‐infective agents.     

The American Isolates: The Little Races

In order to bring the study of bi- and tri-mixed racial groups in America and elsewhere into the general body of racial theory, it is suggested that such groups be considered "little races" From this point of view, considerations having to do with "racialization" and "de-racialization" may find a place in a larger frame of reference along with tribalization, detribalization, and processes leading to group differentiation generally. The little races are marginal groups, but not necessarily aggregations of marginal men. They are made aware of themselves as marginal groups as they become objects of scurrilous epithets thrown at them by larger neighboring racial groups. This leads to a search for a name of dignity, a name inherited from "pure" and honorable ancestors. A number of such origin myths are mentioned. Modern ecological, economic, political, and social changes threaten the future of the little races.