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.

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.

Inhibition of angiogenesis by recombinant human platelet factor-4 and related peptides.

Recombinant human platelet factor-4 (rhPF4), purified from Escherichia coli, inhibited blood vessel proliferation in the chicken chorioallantoic membrane in a dose-dependent manner. Treatment of several cell types with rhPF4 in vitro suggested that the angiostatic effect was due to specific inhibition of growth factor-stimulated endothelial cell proliferation. The inhibitory activities were associated with the carboxyl-terminal, heparin-binding region of the molecule and could be abrogated by including heparin in the test samples, an indication that sulfated polysaccharides might modulate the angiostatic activity of platelet factor-4 in vivo. Understanding of the mechanisms of control of angiogenesis by endogenous proteins should facilitate the development of effective treatments for diseases of pathogenic neovascularization such as Kaposi's sarcoma, diabetic retinopathy, and malignant tumor growth.

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.

Inhibition of tumor growth in mice by an analogue of platelet factor 4 that lacks affinity for heparin and retains potent angiostatic activity.

An analogue of human platelet factor 4 (PF4) lacking affinity for heparin was specifically designed to evaluate the importance of this property in the antitumor effects of recombinant PF4. The purified protein, recombinant PF4-241 (rPF4-241), failed to bind heparin but retained the ability to suppress the growth of tumors in mice. Daily intralesional injections of rPF4-241 significantly inhibited the growth of the B-16 melanoma in syngeneic mice without direct inhibitory effects on B-16 cell growth in vitro. Similar antitumor effects were observed with the human colon carcinoma, HCT-116, grown in nude mice, indicating that the inhibitory activity was neither tumor-type specific nor T-cell dependent. rPF4-241 inhibited endothelial cell proliferation in vitro with dose dependence similar to the native sequence rPF4. Both rPF4 and rPF4-241 inhibited angiogenesis in the chicken chorioallantoic membrane. The analogue, however, was inhibitory at lower concentrations than rPF4 in the chorioallantoic membrane system and its inhibitory effects were not abrogated by the presence of heparin. The present findings support the conclusion that both rPF4 and rPF4-241 inhibit tumor growth by suppression of tumor-induced neovascularization. The finding that this activity is independent of heparin binding may allow the development of PF4-based angiostatic agents with reduced toxicity and improved bioavailability. These results also suggest that PF4 may play a more specific role in modulation of blood vessel development than previously recognized.

Hemoglobin polymerization in sickle cells studied by circular polarized light scattering.

We have studied intracellular polymerization of hemoglobin S in suspensions of small populations of sickle cells using circular polarized light scattering. We argue that the preferential scattering of right circular polarized light (as expressed by measurements of the S14 Mueller scattering matrix element) directly reflects the amount of polymer inside cells. This technique has made it possible to investigate the effect of oxygen tension, cell density and osmotic stress on intracellular hemoglobin polymerization. Using S14 to determine hemoglobin polymer, we show that the polymer increases with deoxyhemoglobin concentration, that cells containing higher hemoglobin concentrations show significantly more polymer than cells containing less hemoglobin, and that polymerization occurs in sickle-trait cells in hypertonic solutions as the oxygen tension in the suspension is reduced. We also present kinetic measurements of polymerization, including that induced by osmotic shock. Finally, we demonstrate that the total light scattered (S11 Mueller scattering matrix element) that is routinely measured simultaneously with S14 can be used to estimate the percent of reduced (deoxy) Hb in the sample. These experiments demonstrate the potential of this technique to monitor hemoglobin polymerization simultaneously with oxygen dissociation under a wide variety of physiological conditions.

Sexual partners, penetrative sexual partners and HIV risk.

This paper argues that the notion of sexual partners per se is insufficient for estimating levels of HIV risk behaviour or changes in HIV risk over time, even though it is a crucial element of most epidemiological models of HIV. The concept of a penetrative sexual partner (PSP) is introduced as a considerably more accurate measure of HIV risk. Using data from a longitudinal study of 930 homosexually active men in England and Wales, this paper demonstrates that variation in numbers of PSPs (and thus HIV risk) is not related to variation in the gross numbers of sexual partners.

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.

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.

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.

Experimental determinations of Mueller scattering matrices for nonspherical particles.

Measurements have been made to determine all sixteen elements of the Mueller scattering matrix for two types of nonspherical particles. Rounded particles of ammonium sulfate and nearly cubic particles of sodium chloride in the 0.1-1.0-mum size range have been prepared by nebulizing salt water solutions and drying the droplets. Scanning electron micrographs are used to determine size distributions used in Mie calculations of all matrix elements. The expected symmetry of the scattering matrices across the diagonal was confirmed, and the expected eight of the sixteen elements were found to be zero within measurement accuracy. The rounded particles were found accurately to obey Mie theory, while the cubic particles were poorly described by Mie theory for some matrix elements and some angles. Total intensity and linear polarization measurements are presented also for a series of increasing sizes of rounded and cubic particles. A discussion of the effect of nonsphericity on the various matrix elements is given, and applications of these results are given to analysis of particle properties in the laboratory, the clouds of Venus, reflection nebulae, the zodiacal light, and atmospheric particulates.

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.

Reentrant tachycardia in a thin layer of ventricular subepicardium: effects of d-sotalol and lidocaine.

Ventricular tachycardia (VT) was induced by premature stimulation or fast pacing in 14 Langendorff-perfused rabbit hearts whose left ventricular endocardial and intramural cells had been selectively killed by freezing. VTs were caused by apparent reentrant excitation in the surviving thin (1 mm thick) subepicardial layer of anisotropically oriented cells having ostensibly normal membrane characteristics. During VT, 100 microM d-sotalol (seven hearts) or 30 microM lidocaine (seven hearts) was added to the perfusate. Electrophysiological variables were measured before and during drug exposure at both slow (S1 = 300 ms) and fast (S1 = 150-180 ms) pacing rates. Sotalol prevented VT reinduction in six of seven preparations, compared to only two of seven with lidocaine. Lidocaine prolonged the functional refractory period (FRP) and slowed conduction velocity (CV). Lidocaine prolonged the wavelength of the cardiac impulse (= FRP x CV) by 18% at slow rates but reduced it by 21% at fast rates. Sotalol, however, since it increased the FRP without reducing CV, caused wavelength prolongation at both rates (43% at slow rates, 26% at fast rates). Thus, this VT model may provide an important contrast of class I and class III drug action, with the drug effects on wavelength predicting susceptibility to VT induction.

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.

Light scattering by Prorocentrum micans: a new method and results.

Striking light-scattering behavior was observed from a marine dinoflagellate, Prorocentrum micans. Measurements of the angular dependence of the 16 Mueller matrix elements were performed on single cells with a polarization-modulation nephelometer by using a new method for cell immobilization. First the dinoflagellate cells were immobilized in a transparent silica gel containing alcohol, and then a second liquid was diffused into the gel to match the index of refraction of the gel network, thereby producing a transparent support medium that scatters less than one tenth the amount of light scattered by a single cell at 90 degrees . Measurements of scattering by a single cell revealed that all 16 matrix elements were significantly nonzero and different from each other. All matrix elements have an extremely rich, reproducible structure that is highly dependent on cell orientation. The matrix elements symmetrically across the diagonal were not equivalent. Striking features of the measurements are the large peak values of S(13), S(14), and other off-diagonal block elements. We believe that this is the first report of such scattering signals by single, suspended marine microorganisms.

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.

Heterosexual behaviour in a large cohort of homosexually active men in England and Wales.

This paper seeks to outline the range and type of heterosexual behaviour in a large non-clinic cohort of homosexually active males. A cohort of 930 homosexually active men were interviewed as part of a prospective study of the seroprevalence of HIV. The paper indicates that over 60% of these men have had at least one sexual experience with a female and that about 90% of these males have engaged in vaginal intercourse with a female. Twelve per cent of the cohort had sexual contact with females in the last year, and 5% had sex with females, in the month preceding interview. Amongst these behaviourally bisexual males the number of female partners is significantly lower than the number of male partners. However, the proportion of female partners with whom penetration occurs is very much higher than for male partners.