Copy Number Variation Analysis in Single-Suture Craniosynostosis: Multiple Rare Variants Including RUNX2 Duplication in Two Cousins With Metopic Craniosynostosis

Little is known about genes that underlie isolated single-suture craniosynostosis. In this study, we hypothesize that rare copy number variants (CNV) in patients with isolated single-suture craniosynostosis contain genes important for cranial development. Using whole genome array comparative genomic hybridization (CGH), we evaluated DNA from 186 individuals with single-suture craniosynostosis for submicroscopic deletions and duplications. We identified a 1.1 Mb duplication encompassing RUNX2 in two affected cousins with metopic synostosis and hypodontia. Given that RUNX2 is required as a master switch for osteoblast differentiation and interacts with TWIST I, mutations in which also cause craniosynostosis, we conclude that the duplication in this family is pathogenic, albeit with reduced penetrance. In addition, we find that a total of 7.5% of individuals with single-suture synostosis in our series have at least one rare deletion or duplication that contains genes and that has not been previously reported in unaffected individuals. The genes within and disrupted by CNVs in this cohort are potential novel candidate genes for craniosynostosis. (C) 2010 Wiley-Liss, Inc

Photosystem-II D1 protein mutants of Chlamydomonas reinhardtii in relation to metabolic rewiring and remodelling of H-bond network at Q(B) site

Photosystem II (PSII) reaction centre D1 protein of oxygenic phototrophs is pivotal for sustaining photosynthesis. Also, it is targeted by herbicides and herbicide-resistant weeds harbour single amino acid substitutions in D1. Conservation of D1 primary structure is seminal in the photosynthetic performance in many diverse species. In this study, we analysed built-in and environmentally-induced (high temperature and high photon fluency-HT/HL) phenotypes of two D1 mutants of Chlamydomonas reinhardtii with Ala250Arg (A250R) and Ser264Lys (S264K) substitutions. Both mutations differentially affected efficiency of electron transport and oxygen production. In addition, targeted metabolomics revealed that the mutants undergo specific differences in primary and secondary metabolism, namely, amino acids, organic acids, pigments, NAD, xanthophylls and carotenes. Levels of lutein, beta-carotene and zeaxanthin were in sync with their corresponding gene transcripts in response to HT/HL stress treatment in the parental (IL) and A250R strains. D1 structure analysis indicated that, among other effects, remodelling of H-bond network at the Q(B) site might underpin the observed phenotypes. Thus, the D1 protein, in addition to being pivotal for efficient photosynthesis, may have a moonlighting role in rewiring of specific metabolic pathways, possibly involving retrograde signalling

Sexual Signalling in Propithecus verreauxi: Male “Chest Badge” and Female Mate Choice

Communication, an essential prerequisite for sociality, involves the transmission of signals. A signal can be defined as any action or trait produced by one animal, the sender, that produces a change in the behaviour of another animal, the receiver. Secondary sexual signals are often used for mate choice because they may inform on a potential partner's quality. Verreaux's sifaka (Propithecus verreauxi) is characterized by the presence of two different morphs of males (bimorphism), which can show either a stained or clean chest. The chest becomes stained by secretions of the sternal gland during throat marking (rubbing throat and chest on a vertical substrate while smearing the scent deposition). The role of the chest staining in guiding female mate choice was previously hypothesized but never demonstrated probably due to the difficulty of observing sifaka copulations in the wild. Here we report that stained-chested males had a higher throat marking activity than clean-chested males during the mating season, but not during the birth season. We found that females copulated more frequently with stained-chested males than the clean-chested males. Finally, in agreement with the biological market theory, we found that clean-chested males, with a lower scent-releasing potential, offered more grooming to females. This “grooming for sex” tactic was not completely unsuccessful; in fact, half of the clean-chested males copulated with females, even though at low frequency. In conclusion, the chest stain, possibly correlated with different cues targeted by females, could be one of the parameters which help females in selecting mates

Post-Exposure Prophylaxis for HIV in the SOF Environment

Special Operations Forces (SOF), both medical and non-medical, have the opportunity to operate in isolated areas while being exposed to a variety of infectious agents. Although the incidence of human immunodeficiency virus (HIV) in the United States military deployed population can be assumed to be nearly zero, experience has shown that our forces are often exposed to populations where the HIV rates can be extremely high or unknown and personal protective measures may not be available. Compliance with the Centers for Disease Control and Prevention guidelines for HIV post-exposure prophylaxis (PEP) is the policy in place for most if not all major military commands. These recommendations are predicated on the fact that the highly active antiretroviral therapy is available in the proposed time frame. Using the Air Force as an example, this article looks at availability of anti-HIV medications on allowance standards. Of a possible 133 allowance standards listed with the United States Air Force Medical Logistics Organization (AFMLO), only 15 (11.27%) contain one or more of the available antiretroviral medications at the time of review. Although these highly active antiretroviral therapy (HAART) medications are not very heat stable and may lose effectiveness when exposed to temperature extremes, it is possible for an individual medic to carry enough medicine to comply with CDC recommendations without significant increase in “weight and cube” packing allowances

A whole cell optical bioassay for the detection of chemical warfare mustard agent simulants

Chemical warfare agents (CWAs) have become a crucial concern of the 20th century in the safety vision of the society and the environment, triggering a growing awareness for early alarm in case of terroristic attacks as well as preventing contamination. Herein, we present the development of a whole-cell optical bioassay for the detection of CWA simulants. In particular, the effects of the mustard agent simulants bis-2-chloroethyl amine and 2-chloroethyl ethyl sulphide on the unicellular green photosynthetic algae Chlamydomonas reinhardtii was studied, with the aim to unravel the response of the microorganism to the presence of these simulants and optimise the analytical conditions of the bioassay. Important variations in the growth, photosynthetic activity, and content of photosynthetic pigments were observed in the presence of the selected simulants. The algal response towards bis-2-chloroethyl amine and 2-chloroethyl ethyl sulphide in a concentration range between 0.2 and 2.5 mM was analysed, indicating a linear relationship in the measured dose-response curves and detection limits of 50 and 200 mu M, respectively. Interference studies demonstrated the suitability of the proposed optical bioassay to detect mustard agent simulants also in drinking water, a defenceless matrix in case of terroristic attack, where atrazine, copper, and arsenic could be present at safety limits. Very slight matrix effect was evidenced, with ratios of 1.04 and 0.86 for calibration curves of bis-2-chloroethyl amine and 2-chloroethyl ethyl sulfide in tap water samples with respect to curves of standard solutions. Recovery values of 104 +/- 15% and 97.5 +/- 6.5% for 1 mM and 2 mM of bis-2-chloroethyl amine and 93 +/- 16% and 105 +/- 5% for 1 mM and 2 mM of 2-chloroethyl ethyl sulfide were achieved. (c) 2017 Elsevier B.V. All rights reserved

gamma-Aminobutyric acidA receptor heterogeneity in rat central nervous system: studies with clonazepam and other benzodiazepine ligands.

none7The properties of [3H]clonazepam, [3H]diazepam and [3H]zolpidem (N,N,6[trimethyl-2-(4-methyl-phenyl)imidazo[1,2-a]pyridine-3-acetamide hemitratrate) binding to synaptic membranes of cerebellum, cortex, olfactory bulb, striatum and spinal cord of rat were compared to the binding properties of [3H]flunitrazepam, [3H]flumazenil and [3H]midazolam. In the cerebellar, cortical and olfactory bulb membranes, the density of high-affinity binding sites of all these tritiated benzodiazepine (BZ) ligands is almost identical. In contrast, in the striatum, the density of [3H]clonazepam and [3H]zolpidem binding sites is approximately 60 and 30%, respectively, of the density of [3H]diazepam, [3H]flunitrazepam or [3H]flumazenil sites. In spinal cord membranes, the number of high-affinity binding sites of [3H]clonazepam and [3H]zolpidem is less than 20% of the number of binding sites for [3H]diazepam, [3H]flunitrazepam, [3H]flumazenil and [3H]midazolam. Moreover, the displacement of [3H]flunitrazepam from spinal cord membranes by clonazepam and zolpidem was characterized by high IC50 values and Hill slopes significantly less than 1. Because [3H]BZ ligand binding in the spinal cord is enhanced by gamma-aminobutyric acid (GABA), these data suggest that different regions of the rat central nervous system may contain different GABA-BZ receptor subtypes. The different pharmacological properties of clonazepam, diazepam and zolpidem (i.e., regarding their ability to enhance bicuculline seizure threshold, to decrease locomotor activity, to induce ataxia or to elicit anticonflict action) further support the concept that in the rat central nervous system preferential occupancy of heterogeneous GABAA receptors by these drugs can be related to their effects on behavior.noneMassotti M;Schlichting JL;Antonacci MD;Giusti P;Memo M;Costa E;Guidotti AMassotti, M; Schlichting, Jl; Antonacci, Md; Giusti, Pietro; Memo, M; Costa, E; Guidotti, A

Design and biophysical characterization of atrazine-sensing peptides mimicking the Chlamydomonas reinhardtii plastoquinone binding niche

The plastoquinone (Q(B)) binding niche of the Photosystem II (PSII) D1 protein is the subject of intense research due to its capability to bind also anthropogenic pollutants. In this work, the Chlamydomonas reinhardtii D1 primary structure was used as a template to computationally design novel peptides enabling the binding of the herbicide atrazine. Three biomimetic molecules, containing the Q(B)-binding site in a loop shaped by two alpha-helices, were reconstituted by automated protein synthesis, and their structural and functional features deeply analysed by biophysical techniques. Standing out among the others, the biomimetic mutant peptide, D1pepMut, showed high ability to mimic the D1 protein in binding both Q(B) and atrazine. Circular dichroism spectra suggested a typical properly-folded alpha-helical structure, while isothermal titration calorimetry (ITC) provided a complete thermodynamic characterization of the molecular interaction. Atrazine binds to the D1pepMut with a high affinity (K-d = 2.84 mu M), and a favourable enthalpic contribution (Delta H = -11.9 kcal mol(-1)) driving the interaction. Fluorescence spectroscopy assays, in parallel to ITC data, provided hyperbolic titration curves indicating the occurrence of a single atrazine binding site. The binding resulted in structural stabilisation of the D1pepMut molecule, as suggested by atrazine-induced cooperative profiles for the fold-unfold transition. The interaction dynamics and the structural stability of the peptides in response to the ligand were particularly considered as mandatory parameters for biosensor/biochip development. These studies paved the way to the set-up of an array of synthetic mutant peptides with a wide range of affinity towards different classes of target analytes, for the development of optical nanosensing platforms for herbicide detection