Microflora of the penile skin-lined neovagina of transsexual women

Background  

The microflora of the penile skin-lined neovagina in male-to-female transsexuals is a recently created microbial niche which thus far has been characterized only to a very limited extent. Yet the knowledge of this microflora can be considered as essential to the follow-up of transsexual women. The primary objective of this study was to map the neo-vaginal microflora in a group of 50 transsexual women for whom a neovagina was constructed by means of the inverted penile skin flap technique. Secondary objectives were to describe possible correlations of this microflora with multiple patients' characteristics, such as sexual orientation, the incidence of vaginal irritation and malodorous vaginal discharge.     

Agglutinating Activity and Structural Characterization of Scalarin,the Major Egg Protein of the Snail Pomacea scalaris (d’Orbigny, 1832)

Apple snail perivitellins are emerging as ecologically important reproductive proteins. To elucidate if the protective functions of the egg proteins of Pomacea canaliculata (Caenogastropoda, Ampullariidae), involved in embryo defenses, are present in other Pomacea species we studied scalarin (PsSC), the major perivitellin of Pomacea scalaris. Using small angle X-ray scattering, fluorescence and absorption spectroscopy and biochemical methods, we analyzed PsSC structural stability, agglutinating activity, sugar specificity and protease resistance. PsSC aggluttinated rabbit, and, to a lesser extent, human B and A erythrocytes independently of divalent metals Ca²⁺ and Mg²⁺ were strongly inhibited by galactosamine and glucosamine. The protein was structurally stable between pH 2.0 to 10.0, though agglutination occurred only between pH 4.0 to 8.0 (maximum activity at pH 7.0). The agglutinating activity was conserved up to 60°C and completely lost above 80°C, in agreement with the structural thermal stability of the protein (up to 60°C). PsSC was able to withstand in vitro gastrointestinal digestion, and showed no trypsin inhibition activity. The presence of lectin activity has been reported in eggs of other Pomacea snails, but here we link for the first time, this activity to an apple snail multifunctional perivitellin. This novel role for a snail egg storage protein is different from closely related P.canaliculata defensive proteins.     

Perspective: Evolution and detection of genetic robustness

Abstract Robustness is the invariance of phenotypes in the face of perturbation. The robustness of phenotypes appears at various levels of biological organization, including gene expression, protein folding, metabolic flux, physiological homeostasis, development, and even organismal fitness. The mechanisms underlying robustness are diverse, ranging from thermodynamic stability at the RNA and protein level to behavior at the organismal level. Phenotypes can be robust either against heritable perturbations (e.g., mutations) or nonheritable perturbations (e.g., the weather). Here we primarily focus on the first kind of robustness–genetic robustness–and survey three growing avenues of research: (1) measuring genetic robustness in nature and in the laboratory; (2) understanding the evolution of genetic robustness; and (3) exploring the implications of genetic robustness for future evolution.     

Nef proteins from simian immunodeficiency virus-infected chimpanzees interact with p21-activated kinase 2 and modulate cell surface expression of various human receptors

The accessory Nef protein allows human immunodeficiency virus type 1 (HIV-1) to persist at high levels and to cause AIDS in infected humans. The function of HIV-1 group M subtype B nef alleles has been extensively studied, and a variety of in vitro activities believed to be important for viral pathogenesis have been established. However, the function of nef alleles derived from naturally simian immunodeficiency virus (SIV)-infected chimpanzees, the original host of HIV-1, or from the HIV-1 N and O groups resulting from independent zoonotic transmissions remains to be investigated. In the present study we demonstrate that SIVcpz and HIV-1 group N or O nef alleles down-modulate CD4, CD28, and class I or II MHC molecules and up-regulate surface expression of the invariant chain (Ii) associated with immature major histocompatibility complex (MHC) class II. Furthermore, the ability of Nef to interact with the p21-activated kinase 2 was generally conserved. The functional activity of HIV-1 group N and O nef genes did not differ significantly from group M nef alleles. However, SIVcpz nef genes as a group showed a 1.8- and 2.0-fold-higher activity in modulating CD28 (P = 0.0002) and Ii (P = 0.016) surface expression, respectively, but were 1.7-fold less active in down-regulating MHC class II molecules (P = 0.006) compared to HIV-1 M nef genes. Our finding that primary SIVcpz nef alleles derived from naturally infected chimpanzees modulate the surface expression of various human cellular receptors involved in T-cell activation and antigen presentation suggests that functional nef genes helped the chimpanzee virus to persist efficiently in infected humans immediately after zoonotic transmission.     

Responses to drought preconditioning in<Emphasis Type="Italic"> Eucalyptus globulus</Emphasis> Labill. provenances

Shoot water relations and morphological responses to drought preconditioning were studied by subjecting 5-month-old seedlings of three provenances of Eucalyptus globulus to different water regimes for 36 days in a greenhouse pot study. Moderately stressed plants were watered every 6 days and severely stressed plants were watered every 9 days. Control plants were watered daily. Drought cycles induced significant changes in morphological and physiological characteristics. Preconditioned seedlings were smaller in size, root collar diameter, height, and leaf area than control seedlings. Shoot/root ratio was not affected by drought. Osmotic potential at full turgor (ψπ_FT) and osmotic potential at turgor loss point (ψπ_TLP) were significantly lower and the magnitude of osmotic adjustment was significantly higher under the severe than under the moderate stress treatment. In severely stressed plants a decrease of turgid mass/dry mass contributed to osmotic adjustment. In a subsequent acclimation test, preconditioned seedlings showed higher values of stomatal conductance, predawn relative water content and water potential and lower mortality than control plants. These variables were significantly related to ψπ_FT. We assume that the reduced leaf area and osmotic adjustment observed in preconditioned seedlings contributed to drought acclimation in the selected E. globulus provenances leading to better rates of gas exchange and improved water status than non-conditioned plants. Provenances exhibited differences in their responses to drought, albeit mainly morphological differences. E. globulus subsp. bicostata from Tumbarumba grew more quickly (larger diameter and height relative growth rate) than the other provenances, implying a greater ability to tolerate water stress. It can be expected that preconditioned seedlings will display greater tolerance of water stress than non-conditioned plants and perform better during early establishment (higher survival and early growth).     

Biology of Cotesia ( = Apanteles) telengai (Hymenoptera: Braconidae) on its natural host Agrotis segetum (Lepidoptera: Noctuidae)

The host suitability of Agrotis segetum Denis & Schiff., A. ipsilon Hufn., Spodoptera littoralis Boisd, S. exigua Hub., Mythimna loreyi Duponchel and Mamestra oleracea L. for the gregarious braconid Cotesia ( = Apanteles) telengai Tobias was determined under laboratory conditions. The parasitoid only completed its development in larvae of A. segetum. The percentage of successfully parasitized larvae and the mean duration of C. telengai egg‐larval period were inversely related to the age of the host or host instar at parasitization. The mean number of parasitoids which emerged per parasitized larva was positively correlated with the larval age. The sex ratio was consistently high (ca. eight males to one female), independent of the host instar parasitized, as compared to 1:1 as observed frequently in field populations of this wasp. The females of C. telengai were active and produced offspring at temperatures of 15, 20, 25 and 30° C. However, the mean percentage of parasitized larvae increased from 13.1 to 72% and the mean progeny per parasitoid female increased from 14.7 to 129.4 parasitoids, both significant, when the experimental temperature was raised from 15 to 30°C, while their mean development time decreased from 75.5 to 19.2 days. At 25°C, the virgin and mated females continued oviposition until days 16 and 17, with a lifetime total of progeny of 397.6 (SD ±224.7) and 611.1 (SD± 128.8) parasitoids respectively, reaching a maximum of 64.3 and 99.2 on day 2 respectively.     

Analysis of Pulsatile Retinal Movements by Spectral-Domain Low-Coherence Interferometry: Influence of Age and Glaucoma on the Pulse Wave

Recent studies have shown that ocular hemodynamics and eye tissue biomechanical properties play an important role in the pathophysiology of glaucoma. Nevertheless, better, non-invasive methods to assess these characteristics in vivo are essential for a thorough understanding of degenerative mechanisms. Here, we propose to measure ocular tissue movements induced by cardiac pulsations and study the ocular pulse waveform as an indicator of tissue compliance. Using a novel, low-cost and non-invasive device based on spectral-domain low coherence interferometry (SD-LCI), we demonstrate the potential of this technique to differentiate ocular hemodynamic and biomechanical properties. We measured the axial movement of the retina driven by the pulsatile ocular blood flow in 11 young healthy individuals, 12 older healthy individuals and 15 older treated glaucoma patients using our custom-made SD-OCT apparatus. The cardiac pulse was simultaneously measured through the use of an oximeter to allow comparison. Spectral components up to the second harmonic were obtained and analyzed. For the different cohorts, we computed a few parameters that characterize the three groups of individuals by analyzing the movement of the retinal tissue at two locations, using this simple, low-cost interferometric device. Our pilot study indicates that spectral analysis of the fundus pulsation has potential for the study of ocular biomechanical and vascular properties, as well as for the study of ocular disease.     

Reaction of Pseudomonas fluorescens kynureninase with beta-benzoyl-L-alanine: detection of a new reaction intermediate and a change in rate-determining step

Beta-benzoyl-DL-alanine was synthesized from alpha-bromoacetophenone and diethyl acetamidomalonate. The racemic amino acid was resolved by carboxypeptidase A-catalyzed hydrolysis of the N-trifluoroacetyl derivative. Beta-benzoyl-L-alanine is a good substrate of kynureninase from Pseudomonas fluorescens, with k(cat) and k(cat)/K(m) values of 0.7 s(-1) and 8.0 x 10(4) M(-1) s(-1), respectively, compared to k(cat) = 16.0 s(-1) and k(cat)/K(m) = 6.0 x 10(5) M(-1) s(-1) for L-kynurenine. In contrast to the reaction of L-kynurenine, beta-benzoyl-L-alanine does not exhibit a significant solvent isotope effect on k(cat) ((H)k/(D)k = 0.96 +/- 0.06). The pre-steady-state kinetics of the reaction of beta-benzoyl-L-alanine were investigated by rapid scanning stopped-flow spectrophotometry. The spectra show the formation of a quinonoid intermediate, with lambda(max) = 490 nm, in the dead time of the instrument, which then decays, with k = 210 s(-1), to form a transient intermediate with lambda(max) at 348 nm. In the presence of benzaldehyde, the 348 nm intermediate decays, with k = 0.7 s(-1), to form a quasistable quinonoid species with lambda(max) = 492 nm. Previous studies demonstrated that benzaldehyde can trap an enamine intermediate formed after the C(beta)-C(gamma) bond cleavage [Phillips, R. S., Sundararaju, B., and Koushik, S. V. (1998) Biochemistry 37, 8783-8789]. Thus, the 348 nm intermediate is kinetically competent. The position of the absorption maximum and shape of the band is consistent with a PMP-ketimine intermediate. The results from chemical quenching analysis do not show a burst of benzoate and, thus, also support the formation of benzoate as the rate-determining step. These data suggest that, in contrast to L-kynurenine, for which the rate-determining step was shown to be deprotonation of the pyruvate-ketimine intermediate [Koushik, S. V., Moore, J. A., III, Sundararaju, B., and Phillips, R. S. (1998) Biochemistry 37, 1376-1382], the rate-determining step in the reaction of beta-benzoyl-L-alanine with kynureninase is C(beta)-C(gamma) bond cleavage.     

Nitrogen dioxide solubility and permeation in lipid membranes

Nitrogen dioxide (NO₂) is an important oxidant molecule in biology that is produced by several biological processes, and it is also an important air pollutant. It can oxidize proteins and lipids with important consequences on their biological functions. Despite its relevance, the interaction of NO₂ with the cell barrier, the lipid membrane, is poorly understood. For instance, can lipid membranes limit NO₂ diffusion? To estimate the permeability of lipid membranes to NO₂ it is necessary to learn more about its solubility in the lipid phase. However, experimental data on NO₂ solubility is very limited. To improve our knowledge on this matter, we used a mixed approach consisting in calculating the solubility of NO₂ and related diatomic and triatomic gases (NO, O₂, CO₂, etc.) in different solvents using quantum calculations and Tomasi’s Polarizable Continuum Model and validating and correcting these results using experimental data available for the related gases. This approach led to an estimated partition coefficient for NO₂ of 2.7 between n-octanol and water, and 1.5 between lipid membranes and water, meaning that NO₂ is a moderately hydrophobic molecule (less than NO, more than CO₂). Based on the solubility-diffusion permeability theory, the permeability coefficient was estimated to be 5 cm s⁻¹, up to 4000 times higher than that of peroxynitrous acid. It is concluded that lipid membranes are not significant barriers to NO₂ transport.