Clinical trial protocol to evaluate the efficacy of cefixime in the treatment of early syphilis.

BACKGROUND: Syphilis rates have been increasing both in the USA and internationally with incidence higher among men-who-have-sex-with-men and people living with human immunodeficiency virus (HIV) infection. Currently, benzathine penicillin is the recommended treatment for syphilis in all patients. Global shortages and cost increases in benzathine penicillin call for alternative treatment options. This study evaluates the efficacy of oral cefixime for the treatment of early syphilis. METHODS: We are conducting a randomized, multisite, open-label, non-comparative clinical trial in Los Angeles and Oakland, CA. Eligible participants are ≥ 18 years old, with primary, secondary, or early latent syphilis (rapid plasma reagin [RPR] titer ≥ 1:8). Patients with HIV infection must have a viral load ≤ 200 copies/mL and CD4+ T cell count ≥ 350 cells/µL during the past 6 months. Participants are randomized to receive either 2.4 M IU benzathine penicillin G intramuscularly once or cefixime 400 mg orally twice a day for 10 days. Participants return at 3, 6, and 12 months post-treatment for follow-up RPR serological testing. The primary outcome is the proportion of participants who achieve ≥ 4-fold RPR titer decrease at 3 or 6 months post-treatment. DISCUSSION: Clinical trials evaluating the efficacy of alternative antibiotics to penicillin are urgently needed. TRIAL REGISTRATION: Clinicaltrials.gov NCT03660488 . Registered on 4 September 2018.

Musculoskeletal problems associated with lameness and poor performance in cutting horses: 200 cases (2007-2015).

OBJECTIVE To describe the chief complaints by owners and the types and prevalences of musculoskeletal problems associated with lameness or poor performance in cutting horses. DESIGN Retrospective case series. ANIMALS 200 client-owned cutting horses examined at the Texas A&M University Veterinary Medical Teaching Hospital between January 1, 2007, and December 31, 2015, because of lameness or poor performance. PROCEDURES Medical records were reviewed, and data were collected regarding signalment, history, findings on physical and lameness examinations, results of diagnostic procedures performed, diagnosis, and treatment. Distribution of observed proportions of forelimb and hind limb involvement was compared with a hypothetical distribution of 50% by means of a χ2 test. RESULTS More horses were examined because of a recent decrease in performance (116/200 [58%]) than for lameness (84 [42%]). All horses had at least 1 lame limb, with lameness affecting a total of 281 limbs. Of the 281 lame limbs, 189 (67%) were hind limbs and 92 (33%) were forelimbs. These proportions were substantially different from a hypothetical distribution of 50% hind limbs and 50% forelimbs. The most common performance change was that horses would not reverse direction to follow prespecified individual cattle, and the most common cause of lameness was pain localized to the stifle joint region (69 [35%]). CONCLUSIONS AND CLINICAL RELEVANCE Cutting horses sustained more hind limb than forelimb musculoskeletal problems, and although these horses were more likely to be examined for decreased performance than lameness, veterinarians should be vigilant for problems affecting the stifle joint region.

Descriptive analysis and early-stage consumer acceptance of yogurts fermented with carrot juice.

This research explored the sensory characteristics and consumer acceptance of novel probiotic unsweetened yogurts. Yogurts were made with 4 carrot juice levels (8, 16, 24, and 32%), 2 firmness levels (regular, 45g/L milk solids; firm, 90g/L milk solids), and 2 starter cultures (C1, C2). The sensory profile characterized the color intensity (before and after stirring), carrot flavor, sourness, and 7 texture/mouth-feel attributes (astringency, chalkiness, mouth-coating, thickness, smoothness, creaminess, and graininess). The influence of carrot juice level and firmness level were evaluated using ANOVA, polynomial contrasts, and principal component analysis. Mean scores and standard errors were calculated. Consumer acceptance panels in Wolfville, Nova Scotia (n=56), and in Vancouver, British Columbia (Asian n=72, non-Asian n=72), evaluated the hedonic responses to the C1 and C2 formulations, respectively. We observed increases in color intensity, carrot flavor, creaminess, mouth-coating, and chalkiness with increasing carrot juice levels, as well as increases in color intensity, carrot flavor, creaminess, mouth-coating, thickness, and astringency with increasing milk solids concentrations of the C1 and C2 yogurts. Mean hedonic scores for color, appearance, and texture/mouth-feel were greater than hedonic scores for aroma, flavor/taste, and overall liking. This research identified the sensory qualities that need further development and demonstrated the importance of early-stage consumer acceptance research for directing new product development.

Development of vocal tract length during early childhood: a magnetic resonance imaging study.

Speech development in children is predicated partly on the growth and anatomic restructuring of the vocal tract. This study examines the growth pattern of the various hard and soft tissue vocal tract structures as visualized by magnetic resonance imaging (MRI), and assesses their relational growth with vocal tract length (VTL). Measurements on lip thickness, hard- and soft-palate length, tongue length, naso-oro-pharyngeal length, mandibular length and depth, and distance of the hyoid bone and larynx from the posterior nasal spine were used from 63 pediatric cases (ages birth to 6 years and 9 months) and 12 adults. Results indicate (a) ongoing growth of all oral and pharyngeal vocal tract structures with no sexual dimorphism, and a period of accelerated growth between birth and 18 months; (b) vocal tract structure's region (oral/anterior versus pharyngeal/posterior) and orientation (horizontal versus vertical) determine its growth pattern; and (c) the relational growth of the different structures with VTL changes with development-while the increase in VTL throughout development is predominantly due to growth of pharyngeal/posterior structures, VTL is also substantially affected by the growth of oral/anterior structures during the first 18 months of life. Findings provide normative data that can be used for modeling the development of the vocal tract.

Efficient gene delivery and gene expression in zebrafish using the Sleeping Beauty transposon.

We used the Tc1/mariner family transposable element Sleeping Beauty (SB) for transgenesis and long-term expression studies in the zebrafish (Danio rerio), a popular organism for clinical disease, vertebrate patterning, and cell biology applications. SB transposase enhanced the transgenesis and expression rate sixfold (from 5 to 31%) and more than doubled the total number of tagged chromosomes over standard, plasmid injection-based transgenesis methods. Molecular analysis of these loci demonstrated a precise integration of these elements into recipient chromosomes with genetic footprints diagnostic of transposition. GFP expression from transposase-mediated integrants was Mendelian through the eighth generation. A blue-shifted GFP variant (BFP) and a red fluorescent protein (DsRed) were also useful transgenesis markers, indicating that multiple reporters are practical for use with SB in zebrafish. We showed that SB is suitable for tissue-specific transgene applications using an abbreviated gamma-crystallin GFP cassette. Finally, we describe a general utility transposon vector for chromosomal engineering and molecular genetics experiments in zebrafish. Together, these data indicate that SB is an efficient tool for transgenesis and expression in zebrafish, and that the transposon will be useful for gene expression in cell biology applications as well as an insertional mutagen for gene discovery during development.

Severe carpometacarpal osteoarthritis in older Arabian horses.

OBJECTIVE: To report a severe form of carpometacarpal osteoarthritis (CMC-OA) affecting primarily older Arabian horses. STUDY DESIGN: Retrospective study. ANIMALS: Thirty-one horses with CMC-OA. METHODS: Carpal radiographs (405 sets) from 3 hospitals were evaluated to identify horses with marked osteoproliferative reaction across the medial aspect of the CMC joint. Owners were contacted to obtain detailed histories and status updates. Necropsy specimens of the CMC joint were evaluated at 2 hospitals to determine the prevalence of 2 variations in the articulation between the proximal second and third metacarpal bones. RESULTS: Thirty-one horses were identified as having marked osteoproliferative reaction at the CMC joint. Twenty-three (74%) were Arabian horses. Of the Arabian horses, the average age at admission was 14.4 years. Eight (34.8%) Arabian horses had a known history of trauma. Most were no longer rideable at presentation. Ten of the horses were subsequently euthanatized because of lameness. The dorsal and palmar articulations between the second and third metacarpal bones were examined in 177 horses. The palmar articulation was absent in 48% of Arabian horses and 12.5% of non-Arabian horses at 1 center, including 4 horses with CMC-OA. At the second center, the palmar articulation was present in 8 of 8 Arabian horses but was absent in 22 of 92 (24%) non-Arabian horses. CONCLUSIONS: An increased frequency of this crippling form of OA was observed in Arabian horses. It may reflect an increased prevalence, in some geographical regions, of an absent palmar articulation between the second and third metacarpal bones. CLINICAL RELEVANCE: Carpal trauma in some Arabian horses may result in unexpectedly severe carpometacarpal osteoarthritis.

Trp203 mutation in GroEL promotes a self-association reaction: a hydrodynamic study.

A combination of sedimentation equilibrium and sedimentation velocity in the analytical ultracentrifuge is used to investigate the hydrodynamic integrity and increased self-association interactions of the mutant GroEL Y203W when compared to the wild-type GroEL molecule, which may be derived from increased hydrophobic exposure caused by the mutation. Sedimentation velocity has revealed that three distinct species were present throughout the concentration ranges used, corresponding to 14-mer (GroEL "super monomer") and 28-mer ("super dimer") subunit compositions with a small amount of 42-mer ("super trimer"), which, from the relative concentration of each species, would give an estimated weight average molecular weight of (1.0 +/- 0.1) x 10(6) Da. Sedimentation equilibrium gave an apparent weight average molecular weight (Mw,app) of (910,000 +/- 5000) Da, which is in agreement with these findings. These results are in contrast to wild-type GroEL which, in excellent agreement with the previous findings of Behlke and co-workers, revealed a single species with an Mw,app of (805,000 +/- 5200) Da and a sedimentation coefficient s(0)20,w of (21.6 +/- 0.3) S. We therefore conclude that the tryptophan mutation at the Y203 location causes a significant degree of self-association of the GroEL 14-mer assembly (with dimer and trimer present). These findings would appear to correlate well with the findings of Gibbons et al., who showed an increase in hydrophobic exposure due to this mutation.

Domain rotations between open, closed and bullet-shaped forms of the thermosome, an archaeal chaperonin.

Three conformations of the thermosome, an archaeal group II chaperonin, have been determined by cryo-electron microscopy (EM). We describe an open form of the double-ring oligomer, a closed form and a bullet-shaped form with one ring open and the other closed. Domain movements have been deduced by docking atomic coordinates into the EM maps. The subunit apical domains, bearing the putative substrate binding sites, rotate about 30 degrees upwards and twist in the plane of the ring from the closed to the open conformation. The closed rings have their nucleotide binding pockets closed by the intermediate domains, but in the open rings, the pocket is accessible.

A kinetic analysis of the nucleotide-induced allosteric transitions of GroEL.

Single-point mutants of GroEL were constructed with tryptophan replacing a tyrosine residue in order to examine nucleotide-induced structural transitions spectrofluorometrically. The tyrosine residues at positions 203, 360, 476 and 485 were mutated. Of these, the probe at residue 485 gave the clearest fluorescence signals upon nucleotide binding. The probe at 360 reported similar signals. In response to the binding of ATP, the indole fluorescence reports four distinct structural transitions occurring on well-separated timescales, all of which precede hydrolysis of the nucleotide. All four of these rearrangements were analysed, two in detail. The fastest is an order of magnitude more rapid than previously identified rearrangements and is proposed to be a T-to-R transition. The next kinetic phase is a rearrangement to the open state identified by electron cryo-microscopy and this we designate an R to R* transition. Both of these rearrangements can occur when only a single ring of GroEL is loaded with ATP, and the results are consistent with the occupied ring behaving in a concerted, cooperative manner. At higher ATP concentrations both rings can be loaded with the nucleotide and the R to R* transition is accelerated. The resultant GroEL:ATP14 species can then undergo two final rearrangements, RR*-->[RR](+)-->[RR](#). These final slow steps are completely blocked when ADP occupies the second ring, i.e. it does not occur in the GroEL:ATP7:ADP7 or the GroEL:ATP7 species. All equilibrium and kinetic data conform to a minimal model in which the GroEL ring can exist in five distinct states which then give rise to seven types of oligomeric conformer: TT, TR, TR*, RR, RR*, [RR](+) and [RR](#), with concerted transitions between each. The other eight possible conformers are presumably disallowed by constraints imposed by inter-ring contacts. This kinetic behaviour is consistent with the GroEL ring passing through distinct functional states in a binding-encapsulation-folding process, with the T-form having high substrate affinity (binding), the R-form being able to bind GroES but retaining substrate affinity (encapsulation), and the R*-form retaining high GroES affinity but allowing the substrate to dissociate into the enclosed cavity (folding). ADP induces only one detectable rearrangement (designated T to T*) which has no properties in common with those elicited by ATP. However, asymmetric ADP binding prevents ATP occupying both rings and, hence, restricts the system to the T*T, T*R and T*R* complexes.

Scaleup of a pharmaceutical granulation in planetary mixers.

The scaleup of a pharmaceutical granulation in a series of planetary mixers with increasing bowl capacity between 5 and 200 liters has been studied by applying the classical dimensionless numbers of Power, Reynolds, and Froude to define the power consumption profile of each mixer as a function of the processing conditions in dimensionless form. Experiments were performed using a formulation based on dicalcium phosphate dihydrate containing pregelatinized starch, with water being added at a fixed rate. Samples were taken at different power consumptions and assessed using mixer torque rheometry. Geometrically similar machines gave the same dimensionless relationship, but when similarity was compromised by the use of modified bowls and blades, different relationships occurred. The results show that, for geometrically similar machines, it is possible to calculate the power consumption at a predefined granulation endpoint at any given operating condition at any scale.

Applicability of a scale-up methodology for wet granulation processes in Collette Gral high shear mixer-granulators.

This study investigates the extension of a scale-up methodology based on dimensionless power relationships, to a series of vertical mixer-granulators in which the bowl is removable, i.e. both impeller and chopper blades are mounted on top-driven vertical shafts positioned through the lid. Granulation runs were carried out in several bowl sizes belonging to the series of Collette Gral mixer-granulators, ranging from 8 to 600 l capacities. It was shown that under certain conditions a common scale-up master curve could be drawn from the data gathered for each bowl, thus permitting the use of such a curve for the determination of mixer-granulator power consumption at a defined granulation end-point. The results also helped to clarify the concept of similitude, both geometrical and dynamic, which is implied in the methodology. The importance of wall slippage (as promoted by the insertion of a PTFE lining into the bowls) and batch size are illustrated.

Process control in a high shear mixer-granulator using wet mass consistency: the effect of formulation variables.

This work investigates the relationships between the wet mass consistency/viscosity of samples prepared in a mixer-granulator and physical properties of the dry granules produced from the wet mass; namely, size distribution, bulk density (Hausner ratio), friability, and flow avalanching behavior. The correlation between the consistency of the wet mass and the downstream dry granule properties confirms that consistency is the key parameter to control in wet granulation by mechanical agitation. Variations in the formulation affect the dimensionless power relationship of the mixer-granulator considered; that is, the equivalence between wet mass consistency and mixer net power consumption, which is actually the parameter used to monitor the wet granulation process. The same variations in formulation also affect the relationships between wet mass consistency and dry granule properties.

A methodology for the optimization of wet granulation in a model planetary mixer.

This paper investigates a methodology for the optimization of wet granulation processes in planetary mixers. A model formulation was granulated in a planetary mixer (two different bowl sizes). The wet masses were characterized by their bulk density and consistency (as measured by mixer torque rheometry), and the feasibility of scale-up from one mixer bowl to the other was studied using a dimensionless numbers approach for the estimation of the power consumption at the granulation end point. Both bowls gave the same dimensionless power relationships (a relationship between the power number, Reynolds number, Froude number, and bowl fill ratio), which could therefore be used for calculating the power consumption level when the wet mass achieves its target values of density and consistency, i.e., the point at which granulation should be stopped. It was also shown that batches granulated in different conditions (batch size, blade speed) in two planetary mixers, but presenting similar wet mass characteristics (bulk density and consistency) led to dry granules of similar properties: granule size distribution, density, friability, and flow. This work suggests that it is possible to characterize the wet mass by only two parameters which describe the quality of the downstream granules. The scale-up procedure based on the use of dimensionless numbers was found to be applicable to planetary mixers, provided they give one common dimensionless power relationship.

Asymmetry, commitment and inhibition in the GroE ATPase cycle impose alternating functions on the two GroEL rings.

The ATPase cycle of GroE chaperonins has been examined by transient kinetics to dissect partial reactions in complexes where GroEL is asymmetrically loaded with nucleotides. The occupation of one heptameric ring by ADP does not inhibit the loading of the other with ATP nor does it prevent the consequent structural rearrangement to the "open" state. However, ADP binding completely inhibits ATP hydrolysis in the asymmetric complex, i.e. ATP cannot by hydrolysed when ADP is bound to the other ring. This non-competitive inhibition of the ATPase by ADP is consistent with a ring-switching, or "two-stroke", mechanism of the type: ATP:GroEL --> ADP:GroEL --> ADP:GroEL:ATP --> GroEL:ATP --> GroEL:ADP, i.e. with respect to the GroEL rings, ATP turns over in an alternating fashion. When the ATP-stabilized, "open" state is challenged with hexokinase and glucose, to quench the free ATP, the open state relaxes slowly (0.44 s-1) back to the apo (or closed) conformation. This rate, however, is three times faster than the hydrolytic step, showing that bound ATP is not committed to hydrolysis. When GroES is bound to the GroEL:ATP complex and the system is quenched in the same way, approximately half of the bound ATP undergoes hydrolysis on the chaperonin complex showing that the co-protein increases the degree of commitment. Thus, non-competitive inhibition of ATP hydrolysis, combined with the ability of the co-protein to block ligand exchange between rings has the effect of imposing a reciprocating cycle of reactions with ATP hydrolysing, and GroES binding, on each of the GroEL rings in turn. Taken together, these data imply that the dominant, productive steady state reaction in vivo is: GroEL:ATP:GroES --> GroEL:ADP:GroES --> ATP:GroEL:ADP:GroES --> ATP:GroEL:ADP --> GroES:ATP:GroEL:ADP --> GroES:ATP:GroEL for a hemi-cycle, and that significant inhibi tion of hydrolysis may arise through the formation of a dead-end ADP:GroEL:ATP:GroES complex.

Volume-sensitive taurine efflux from mammary tissue is not obliged to utilize volume-activated anion channels.

Cell-swelling, induced by a hyposmotic shock, activates the release of taurine from lactating rat mammary tissue explants. The degree of stimulation of taurine efflux was dependent upon the extent of cell-swelling. Volume-sensitive taurine release was attenuated by the anion transport inhibitors NPPB, DIOA, DIDS, niflumate, flufenamate, mefenamate and diiodosalicylate but not by salicylate. Cell-swelling, following a hyposmotic challenge, did not increase the unidirectional efflux of radiolabelled I- or D-asparate from mammary tissue explants. The results suggest that although mammary tissue expresses a volume-sensitive amino acid transport system which is inhibited by anion transport blockers the pathway has no identity with volume-activated anion channels.

Sensitization and desensitization to capsaicin and menthol in the oral cavity: interactions and individual differences.

It was reported in a recent study that, like capsaicin, menthol is capable of producing a desensitization to sensory irritation in the oral cavity. Whereas capsaicin is known to be able to cross-desensitize with other chemical irritants, no such information exists for menthol. To address this question, the first experiment was designed to reveal whether cross-desensitization would occur between menthol and capsaicin. After a pretest on the tongue tip in which subjects rated the intensity of irritation and cold produced by 3.5 ppm capsaicin or 0.3% l-menthol, five samples of the same stimuli were sipped and swished at 1-min intervals for 5 min. Fifteen minutes later subjects were tested on the tongue tip with either capsaicin or menthol. The results 1) confirmed self-desensitization for both chemicals, 2) demonstrated cross-desensitization of menthol by capsaicin, and 3) revealed cross-sensitization of capsaicin by menthol. This series of outcomes suggests that menthol produces much of its sensory irritation via capsaicin-sensitive pathways, but that it excites and/or desensitizes those pathways via different mechanisms than does capsaicin. Analysis of the individual data revealed large differences in sensitization, and desensitization that were significantly correlated across chemicals, which suggests the possibility that the perceptual response to repeated exposures to irritants may be idiosyncratic. Contrary to earlier findings, the first experiment also revealed apparent self- and cross-desensitization of the menthol sensation of coolness. The latter outcome was investigated in a second experiment in which the effect of capsaicin desensitization on the perception of physical as well as chemical (menthol) cooling was measured when the stimuli were presented as oral rinses. No desensitization was found for either form of stimulation, which implied the apparent desensitization of coolness in Experiment I may have been due to the difficulty of discriminating sensations of cold from sensations of chemical irritation. The overall findings are discussed in terms of the complex sensory and perceptual interactions that take place within the chemesthetic modality.

Sensory irritation and coolness produced by menthol: evidence for selective desensitization of irritation.

The temporal characteristics of the oral perception of menthol solutions were explored in two experiments. In Experiment 1, 10 samples of either 0.03% or 0.30% menthol were presented at 1 min intervals and rated for the perceived intensity of cooling and irritation. Reports of sensation quality (burning, tingling, stinging and numbing) and pain were also collected. At the higher concentration, a significant decrease in perceived intensity was observed over time for irritation, but not for cooling. Experiment 2 was designed to explore further the nature of the decline in irritation observed in Experiment 1. Employing 1-min and 5-min inter-stimulus intervals between solutions, it was found that the decrease in menthol irritation more closely resembled desensitization than adaptation. Decreases in the frequency of reports of the burning and stinging qualities, but not the tingling, numbing or cooling qualities, suggested that menthol has a specific desensitizing effect on a population of mucosal nociceptive fibers.