Sexual dimorphism of human vallate papillae: an in vivo study of normative morphology.

The perimeters of vallate papillae (VP) house approximately half of the taste buds on the human tongue. However, little information exists regarding perimeter measurements of VP. Likewise, great diversity exists among reports of the number of VP and diameter of VP, in general. The research presents an analysis of the perimeters, counts, and diameters of VP in vivo. Endoscopic examination was performed on 79 individuals (40 females, 39 males) between 18 and 26 years of age. A total of 583 VP were counted, 565 of which were able to be measured. Data revealed a statistically significant difference between male and female VP count (t(75.6) = 4.5; p = 0.00003). Females had, on average, 2.22 more VP than males. Males were found to have larger mean VP diameter per person and mean VP perimeter per person than females (t(58.9) = -2.4; p = 0.021 and t(59.3) = -2.4; p = 0.019, respectively). The report demonstrates that VP are sexually dimorphic at the gross anatomical level.

20-Hydroxyecdysone release from biodegradable devices: the effect of size and shape.

20-Hydroxyecdysone (20-OH) is a natural compound with many demonstrated effects on the physiological functions of vertebrates, particularly increased protein synthesis. Our study sought a suitable dosage form with continuous release of the drug lasting several weeks for implantation into agricultural animals. Biodegradable microparticles and implants of poly(L-lactic) and poly(DL-lactic) acids were prepared. Oligomers of these materials were synthesized, and a method of melting the binary mixture of the oligomer and 20-OH was employed. The particles were prepared simply by grinding the solidified block of the melt and sieving. Implants were prepared by extruding the melt into silicone tubes, removing the solidified content, and cutting into cylinders of 2 mm diameter and various lengths. A new method of preparation of hollow cylinders by aspirating air into silicone tubes filled with the melt was developed. The experiments demonstrated stability of 20-OH during heat treatment. Release of the active ingredient was tested in static in vitro conditions, analogous to those at the site of implantation, and prolonged drug release was obtained with both types of implant. The hollow implants gave release rates nearest to ideal zero-order kinetics and would appear most appropriate for testing in vivo.

The dynamic behavior of individual microtubules associated with chromosomes in vitro.

Mitotic movements of chromosomes are usually coupled to the elongation and shortening of the microtubules to which they are bound. The lengths of kinetochore-associated microtubules change by incorporation or loss of tubulin subunits, principally at their chromosome-bound ends. We have reproduced aspects of this phenomenon in vitro, using a real-time assay that displays directly the movements of individual chromosome-associated microtubules as they elongate and shorten. Chromosomes isolated from cultured Chinese hamster ovary cells were adhered to coverslips and then allowed to bind labeled microtubules. In the presence of tubulin and GTP, these microtubules could grow at their chromosome-bound ends, causing the labeled segments to move away from the chromosomes, even in the absence of ATP. Sometimes a microtubule would switch to shortening, causing the direction of movement to change abruptly. The link between a microtubule and a chromosome was mechanically strong; 15 pN of tension was generally insufficient to detach a microtubule, even though it could add subunits at the kinetochore-microtubule junction. The behavior of the microtubules in vitro was regulated by the chromosomes to which they were bound; the frequency of transitions from polymerization to depolymerization was decreased, and the speed of depolymerization-coupled movement toward chromosomes was only one-fifth the rate of shortening for microtubules free in solution. Our results are consistent with a model in which each microtubule interacts with an increasing number of chromosome-associated binding sites as it approaches the kinetochore.

Activation of the MKK/ERK pathway during somatic cell mitosis: direct interactions of active ERK with kinetochores and regulation of the mitotic 3F3/2 phosphoantigen.

The mitogen-activated protein (MAP) kinase pathway, which includes extracellular signal-regulated protein kinases 1 and 2 (ERK1, ERK2) and MAP kinase kinases 1 and 2 (MKK1, MKK2), is well-known to be required for cell cycle progression from G1 to S phase, but its role in somatic cell mitosis has not been clearly established. We have examined the regulation of ERK and MKK in mammalian cells during mitosis using antibodies selective for active phosphorylated forms of these enzymes. In NIH 3T3 cells, both ERK and MKK are activated within the nucleus during early prophase; they localize to spindle poles between prophase and anaphase, and to the midbody during cytokinesis. During metaphase, active ERK is localized in the chromosome periphery, in contrast to active MKK, which shows clear chromosome exclusion. Prophase activation and spindle pole localization of active ERK and MKK are also observed in PtK1 cells. Discrete localization of active ERK at kinetochores is apparent by early prophase and during prometaphase with decreased staining on chromosomes aligned at the metaphase plate. The kinetochores of chromosomes displaced from the metaphase plate, or in microtubule-disrupted cells, still react strongly with the active ERK antibody. This pattern resembles that reported for the 3F3/2 monoclonal antibody, which recognizes a phosphoepitope that disappears with kinetochore attachment to the spindles, and has been implicated in the mitotic checkpoint for anaphase onset (Gorbsky and Ricketts, 1993. J. Cell Biol. 122:1311-1321). The 3F3/2 reactivity of kinetochores on isolated chromosomes decreases after dephosphorylation with protein phosphatase, and then increases after subsequent phosphorylation by purified active ERK or active MKK. These results suggest that the MAP kinase pathway has multiple functions during mitosis, helping to promote mitotic entry as well as targeting proteins that mediate mitotic progression in response to kinetochore attachment.

Polarized light scattering by aerosols in the marine atmospheric boundary layer.

The intensity and polarization of light scattered from marine aerosols affect visibility and contrast in the marine atmospheric boundary layer (MABL). The polarization properties of scattered light in the MABL vary with size, refractive index, number distributions, and environmental conditions. Laboratory measurements were used to determine the characteristics and variability of the polarization of light scattered by aerosols similar to those in the MABL. Scattering from laboratory-generated sea-salt-containing (SSC) [NaCl, (NH(4))(2) SO(4), and seawater] components of marine aerosols was measured with a scanning polarization-modulated nephelometer. Mie theory with Gaussian and log normal size distributions of spheres was used to calculate the polarized light scattering from various aerosol composition models and from experimentally determined distributions of aerosols in the marine boundary layer. The modeling was verified by comparison with scattering from distilled water aerosols. The study suggests that polarimetric techniques can be used to enhance techniques for improving visibility and remote imaging for various aerosol types, Sun angles, and viewing conditions.

Laboratory studies of angle- and polarization-dependent light scattering in sea ice.

The angle- and polarization-dependent light scattering were measured for oriented first-year and multiyear sea ice taken from the Chukchi Sea near Pt. Barrow, Alaska. The entire Mueller matrix for these samples was determined at 532 nm. Mueller matrices were also determined for artificially grown saline ice samples and melted samples of the respective ice types. Phase functions for thin-slab samples are qualitatively consistent with calculations for scattering from brine inclusions in a solid ice medium and depend strongly on the shape of the scattering sample. Small orientation-dependent effects are observed for scattering from oriented sea ice. A simple model is used to describe qualitatively some features of the measured sea ice Mueller matrices. This model combines the effects of scattering from spherical inhomogeneities and the intrinsic birefringence of pure water ice. A set of Mueller matrix inequalities is presented and used to obtain physical insight into the measurement results.

Patterning of functional antibodies and other proteins by photolithography of silane monolayers.

We have demonstrated the assembly of two-dimensional patterns of functional antibodies on a surface. In particular, we have selectively adsorbed micrometer-scale regions of biotinylated immunoglobulin that exhibit specific antigen binding after adsorption. The advantage of this technique is its potential adaptability to adsorbing arbitrary proteins in tightly packed monolayers while retaining functionality. The procedure begins with the formation of a self-assembled monolayer of n-octadecyltrimethoxysilane (OTMS) on a silicon dioxide surface. This monolayer can then be selectively removed by UV photolithography. Under appropriate solution conditions, the OTMS regions will adsorb a monolayer of bovine serum albumin (BSA), while the silicon dioxide regions where the OTMS has been removed by UV light will adsorb less than 2% of a monolayer, thus creating high contrast patterned adsorption of BSA. The attachment of the molecule biotin to the BSA allows the pattern to be replicated in a layer of streptavidin, which bonds to the biotinylated BSA and in turn will bond an additional layer of an arbitrary biotinylated protein. In our test case, functionality of the biotinylated goat antibodies raised against mouse immunoglobulin was demonstrated by the specific binding of fluorescently labeled mouse IgG.

The force exerted by a single kinesin molecule against a viscous load.

Kinesin is a motor protein that uses the energy derived from the hydrolysis of ATP to power the transport of organelles along microtubules. To probe the mechanism of this chemical-to-mechanical energy transduction reaction, the movement of microtubules across glass surfaces coated with kinesin was perturbed by raising the viscosity of the buffer solution. When the viscosity of the solution used in the low density motility assay was increased approximately 100-fold through addition of polysaccharides and polypeptides, the longer microtubules, which experienced a larger drag force from the fluid, moved more slowly than the shorter ones. The speed of movement of a microtubule depended linearly on the drag force loading the motor. At the lowest kinesin density, where dilution experiments indicated that the movement was caused by a single kinesin molecule, extrapolation of the linear relationship yielded a maximum time-averaged drag force of 4.2 +/- 0.5 pN per motor (mean +/- experimental SE). The magnitude of the force argues against one type of "ratchet" model in which the motor is hypothesized to rectify the diffusion of the microtubule; at high viscosity, diffusion is too slow to account for the observed speeds. On the other hand, our data are consistent with models in which force is a consequence of strain developed in an elastic element within the motor; these models include a different "ratchet" model (of the type proposed by A. F. Huxley in 1957) as well as "power-stroke" models.

Gay men as victims of nonconsensual sex.

Incidents of nonconsensual sexual activity among 930 homosexually active men living in England and Wales are analyzed. Of these men, 27.6% said they had been sexually assaulted or had sex against their will at some point in their lives; one third had been forced into sexual activity (usually anal intercourse) by men with whom they had previously had, or were currently having, consensual sexual activity. The contention that male rape is usually committed by heterosexually identified men, primarily as an expression of power and control, is not supported. Recognition that gay men rape other gay men is needed, both by the gay community and support services for victims.

Determination of the average orientation of DNA in the octopus sperm Eledone cirrhossa through polarized light scattering.

The coupled-dipole approximation has been used to model polarized light-scattering data obtained from the sperm of the octopus Eledone cirrhosa. Mueller scattering-matrix elements (which describe how a sample alters the intensity and degree of polarization of scattered light) were measured as a function of angle. The sample was modeled as a helical fiber believed to correspond to a DNA protein complex. It was necessary to propose an inherent anisotropy in the polarizability of the fiber in order to fit the data. The direction of the principle axes of the polarizability were determined by comparing the model with experimental data. The results suggest that the 2-nm DNA fibers are perpendicular to the thick fiber that defines the helical geometry of the octopus sperm head.

Kinesin swivels to permit microtubule movement in any direction.

Kinesin is a motor protein that uses the energy derived from ATP hydrolysis to transport organelles along microtubules. By analyzing the thermal fluctuation of microtubules tethered to glass surfaces by single molecules of kinesin, we have measured the torsional flexibility of the motor protein. The torsional stiffness of kinesin, (117 +/- 19) x 10(-24) N.m.rad-1 (mean +/- SEM), is so low that one kT of energy (approximately 4.1 x 10(-21) J at room temperature) is sufficient to twist a kinesin molecule through more than 360 degrees from its resting orientation. Consistent with this flexibility, motility assays show that one or more kinesin molecules can move a microtubule equally well in any direction. These results explain how a motor on the surface of an organelle can rapidly bind to and capture a microtubule irrespective of the organelle's orientation. Furthermore, the flexibility ensures that several motors can efficiently work together even though they are randomly oriented on the surface of an organelle rather than being in precise arrays like the motors of muscle and cilia.

Sex role separation in sexual diaries of homosexual men.

OBJECTIVE: To measure types of sex role prevalence in common and risk-related behaviours among gay men for modelling HIV transmission. DESIGN: Cohort study of 385 homosexually active men recording sexual diaries over 1-month periods. METHODS: Measures of incidence of behavioural sex roles for masturbation, fellatio, anal intercourse and anilingus by relationship type, derived from 1-month sexual diary data. RESULTS: Low behavioural role rigidity for masturbation and fellatio, but higher rigidity for anal intercourse and anilingus. Participants with no regular partner showed a relatively low frequency of anal intercourse, whereas those in closed relationships showed a high frequency. CONCLUSION: Although anal intercourse shows a certain degree of behavioural role rigidity, this rigidity is not large enough to conclude that gay men exclusively engage in either an active or a passive role. Typical rates for exclusive active and passive roles for anal intercourse during the month the diaries were recorded were in the range of 12-15%; the dual role was significantly higher.

The testing of saliva samples for HIV-1 antibodies: reliability in a non-clinic setting.

AIMS: To assess the reliability of saliva samples as a means of testing for HIV-antibodies outside clinic settings. METHODS: Men taking part in a non-clinic longitudinal study of homosexually active men provided samples of saliva and blood. Sera were screened using a competitive ELISA (Wellcozyme) and positive sera were confirmed by an indirect ELISA (Abbott). Saliva samples were screened either using an IgG captive radioimmunoassay or an amplified ELISA. RESULTS: A total of 534 paired saliva and blood samples were tested. Overall sensitivity was 96.2% and specificity was 100%. None of the saliva tests were falsely positive for HIV-1 antibodies. CONCLUSIONS: HIV-1 saliva tests can reliably be used in a non-clinic or field setting. However, if results are to be given to respondents, it is necessary to offer adequate counselling and consider the mechanisms for referral and follow-up for those that are found to be HIV-1 antibody positive.

No connection between alcohol use and unsafe sex among gay and bisexual men.

OBJECTIVE: To investigate the relationship between alcohol use and unsafe sexual behaviour. METHODS: The paper discusses data collected from 461 gay and bisexual men interviewed in England and Wales by Project SIGMA during 1991-1992. These data were collected during face-to-face interviews using retrospective weekly diary techniques and include details of all sexual sessions and alcohol use. The 819 reported sexual sessions with other men are divided into those that involved alcohol use (30.6%) and those that did not. RESULTS: Differences in the incidence of HIV risk behaviours between sexual sessions that involved alcohol use and those that did not are small, and none are statistically significant. Furthermore, for those men who engaged in sexual behaviour whilst under the influence of alcohol, the quantity of alcohol consumed had no effect on sexual behaviour. CONCLUSIONS: Among gay and bisexual men, sex under the influence of alcohol is no more likely to be unsafe than sex among men who have not consumed alcohol.

Changes in condom use by gay men.

Cross sectional and longitudinal data on condom use among homosexually active men is presented. Data pertain to two waves of interviewing (1987/8 and 1991) from Project SIGMA, a large longitudinal study of male homosexual behaviour in the light of HIV. Aggregate changes across the time-period indicate an increase in condom use for both insertive and receptive anal intercourse, particularly with casual partners. Condom use is still much more prevalent with casual rather than regular partners. Longitudinal analysis show considerable changes at the individual level, with almost a third of men changing their practices. Half of those decreased their risk behaviours, whilst the other half increased them. Less than 1% used condoms for fellatio in the year preceding the 1991 interview, but 8.3% had used other barriers during sex, primarily for hygiene and fun reasons, rather than as disease prophylaxis. Some implications are drawn for condom promotion and HIV prevention.

PIP: Even though the uptake of condoms by homosexual and bisexual males definitely ranks as one of the most dramatic health protective behavioral changes ever recorded, HIV prevention campaigns nonetheless continue to encourage these men to use condoms during anal intercourse. Maybe 10% of gay men used condoms prior to 1980, and then primarily for reasons of hygiene, but 78% of gay and bisexual men use them for anal intercourse by 1987 and were regularly used by 50% of all men engaging in anal sex. This paper reports changes in condom use in the only non-clinic longitudinal study of gay and bisexual) men in England and Wales. 358 men were interviewed 1987/88 and 1991, and a total of 472 men in 1991. The participants were predominantly well-educated, White, and happy with their sexuality. At the aggregate level, an increase in condom use was observed over the period for both insertive and receptive anal intercourse, especially with casual partners. Longitudinal analysis shows a lot of change at the individual level, with almost a third of the men changing their practices; half of these, however, decreased their risk behaviors, while the other half increased them. Further, less than 1% used condoms for fellatio in the year preceding the 1991 interview, but 8.3% had used other barriers such as rubber gloves and dental dams during sex mainly for hygiene and fun reasons instead of for disease prevention. The authors comment on the implications of these findings for condom promotion and HIV prevention.

Molten globule monomer to condensed dimer: role of disulfide bonds in platelet factor-4 folding and subunit association.

Platelet factor 4 (PF4) exhibits high affinity for heparin and exists as a tetramer in solution under physiologic conditions. Reduction of the two disulfide bridges in PF4 increases the protein's dissociation constant for heparin approximately 20-fold and shifts the highest apparent aggregation state from tetramer to dimer as evidenced by gel filtration, chemical cross-linking, and 1H-NMR studies. 1H-NMR spectra of reduced PF4 monomers generally show narrower, less dispersed, upfield-shifted NH and alpha H resonances, suggesting the presence of an unfolded monomer state. Reduced PF4 monomer folding, however, is evidenced by the presence of about 12 relatively long-lived backbone NHs and by CD spectra that indicate conservation of overall secondary structure. These data suggest the presence of a molten globule-type state. Urea denaturation shifts this apparent molten globule to a fully unfolded state characterized by more random coil-like resonance shifts. The reduced PF4 dimer state yields NMR and CD data consistent with preservation of tertiary structural folds found for the native species. In this regard, the reduced PF4 folding transition is thermodynamically linked with dimer formation which stabilizes tertiary structure. Monomer-dimer association equilibria for reduced PF4 essentially follow the same pH and salt titration trends as reported previously for native PF4 dimers [Mayo, K. H., & Chen, M. J. (1989) Biochemistry 28, 9469-9478], indicating that that dimer interface is generally conserved in the absence of disulfide constraints. Reduced PF4 tetramers are not apparent under any conditions investigated, suggesting that disulfides are necessary for efficient antiparallel beta-sheet alignment between dimer pairs.