Excitons in CsPbBr3 Halide Perovskite.

Excitons in Bridgman grown halide perovskite CsPbBr3 single crystals were examined using photoluminescence (PL) spectroscopy to determine the nature of the electronic states. The photoluminescence intensity was strongly temperature-dependent and depended upon the specific exciton band. At low temperatures intrinsic disorder and its related shallow below bandgap tail states determine the emission properties. Photoluminescence at low temperature revealed the presence of several strong bands at the band edge that is attributed to free or trapped/bound excitons. This PL emission results from strong electron-phonon coupling with an average phonon energy Eph of 6.5 and 27.4 meV for the emissions, comparable to that observed in other perovskites. The Huang-Rhys parameter S was calculated to be 3.81 and 1.51, indicating strong electron-phonon coupling. The interactions between electrons and phonons produce small polarons that tend to bind charge carriers and result in trapped/bound excitons. The transient photoluminescence response of each specific band was studied, and the results indicated a multiphonon recombination process. Average PL lifetimes of ∼17 ns for free excitons and ∼38 ns for trapped/bound excitons were determined. The observed edge states could be associated with native defects such as vacancies and interstitials, as well as twinning due to the cubic-to-tetragonal phase transition in CsPbBr3. Elimination of the trapping sites for binding excitons could lead to improved charge transport mobilities, carrier lifetimes, and detector properties in this system.

Integration of TB-HIV services at an ANC facility in Frances Baard District, Northern Cape, South Africa.

BACKGROUND: Integrated tuberculosis-human immunodeficiency virus (TB-HIV) service delivery as part of maternal health services, including antenatal care (ANC), is widely recommended. This study assessed the implementation of collaborative TB-HIV service delivery at a hospital-based ANC service unit. METHODS: A record review of a random sample of 308 pregnant women attending the ANC service between April 2011 and February 2012 was conducted. Data were extracted from registers and patient case notes. Outcomes included the proportion of women who underwent HIV counselling and testing (HCT), CD4 count testing, antiretroviral treatment (ART), cotrimoxazole preventive treatment (CPT), TB screening and isoniazid preventive treatment (IPT). Analysis measured variations in patient characteristics associated with service delivery. RESULTS: All women underwent HCT; 80% of those who tested HIV-positive were screened for TB. Most (85.9%) of the HIV-positive women received a CD4 count. However, only 12.9% of eligible women received ART prophylaxis onsite, only 35.7% were referred for initiation of ART, only 42.3% commenced IPT and none received CPT or further investigations for TB. HIV-negative women had 2.6 higher odds (95%CI 1.3-5.3) of receiving TB screening than their HIV-positive counterparts. CONCLUSIONS: Although the identification of HIV-positive women and TB suspects was adequate, implementation of other TB-HIV collaborative activities was sub-optimal.

Contexte : Une offre de services intégrée de tuberculose et du virus de l'immunodéficience humaine (TB-VIH)­en tant qu'élément des services de santé maternelle, notamment des consultations prénatales (ANC)­est largement recommandée. Cette étude a évalué la mise en œuvre d'une offre de services intégrée TB-VIH dans une unité hospitalière de CPN.Méthodes : Les dossiers d'un échantillon aléatoire de 308 femmes enceintes qui ont fréquenté le service ANC entre avril 2011 et février 2012 ont été revus. Les données ont été extraites à partir des registres ainsi que des dossiers des patients. Les résultats attendus comprenaient la proportion de femmes bénéficiant d'un conseil et test VIH (HCT), d'un comptage des CD4, d'un traitement antirétroviral (ART), d'un traitement préventif par cotrimoxazole (CPT), d'un dépistage de TB et d'un traitement préventif par isoniazide (IPT). L'analyse a mesuré les variations des caractéristiques des patients associées à l'offre de services.Résultats : Toutes les femmes ont bénéficié du HCT et 80% de celles ayant eu un test VIH positif ont eu un dépistage de TB. La majorité (85,9%) des femmes VIH-positives ont eu un comptage des CD4. Cependant, seulement 12,9% des femmes éligibles ont reçu une prophylaxie ART sur place ; seulement 35,7% ont été référées pour une mise en route de l'ART ; seulement 42.3% ont commencé l'IPT ; et aucune n'a reçu de CPT ni d'autres investigations relatives à la TB. Les femmes VIH négatives avaient 2,6 fois (IC95% 1,3­5,3) plus de chances de bénéficier d'un dépistage de TB que leurs homologues VIH positives.Conclusions: L'identification des femmes VIH positives et de celles suspectes de TB a été satisfaisante, mais la mise en œuvre des autres activités de collaboration TB-VIH a été insuffisante.

Marco de referencia: La prestación integrada de servicios de atención de la tuberculosis (TB) y la infección por el virus de la inmunodeficiencia humana (VIH) se recomienda ampliamente como parte de los servicios que se ofrecen a las madres durante la atención prenatal (ANC). En el presente estudio se evaluó la introducción de los servicios integrados de TB y VIH en una unidad hospitalaria de ANC.Método: Se examinaron las historias clínicas de una muestra aleatoria de 308 embarazadas que acudieron al servicio de ANC entre abril del 2011 y febrero del 2012. Se extrajeron datos de los registros y las historias clínicas de las pacientes. Los criterios de evaluación fueron la proporción de mujeres en quienes se practicó la orientación y las pruebas diagnósticas del VIH (HCT), el recuento de linfocitos CD4, el tratamiento antirretrovírico (ART), el tratamiento preventivo con cotrimoxazol (CPT), la detección sistemática de la TB y el tratamiento preventivo con isoniazida (IPT). En el análisis se midieron las variaciones en las características de las pacientes asociadas con la prestación de los servicios.Resultados: Todas las mujeres recibieron HCT y en 80% que obtuvieron un resultado positivo, se practicó la detección sistemática de la TB. En la mayoría de las pacientes positivas frente al VIH se practicó el recuento de linfocitos CD4 (85,9%). Sin embargo, solo 12,9% de las mujeres aptas recibieron la profilaxis ART en el lugar de la consulta; solo 35,7% se remitieron con el fin de comenzar el ART; apenas 42,3% de las pacientes comenzaron el IPT; y ninguna recibió CPT ni tuvo investigaciones complementarias por TB. Las mujeres con resultados negativos frente al VIH exhibieron un cociente de posibilidades 2,6 veces inferior de beneficiar de la detección sistemática de la TB, en comparación con las mujeres VIH positivas.Conclusión: Se constató una detección adecuada de las mujeres positivas frente al VIH y de casos con presunción clínica de TB, pero una prestación deficiente de las demás actividades de los servicios integrados del VIH y la TB.

Familial testicular germ cell tumors (FTGCT) - overview of a multidisciplinary etiologic study.

This Review summarizes the cumulative results of the National Cancer Institute Clinical Genetics Branch Multidisciplinary Etiologic Study of Familial Testicular Germ Cell Tumors (FTGCT). Initiated 12 years ago, this protocol enrolled 724 subjects from 147 unrelated families with either ≥2 affected men (n = 90) with TGCT or a proband with bilateral TGCT and a negative family history for this cancer (n = 57). Data were collected directly from 162 subjects evaluated at the NIH Clinical Center, and 562 subjects provided information from their home communities (Field Cohort). The primary study aims included (i) ascertaining, enrolling eligible FTGCT kindred, (ii) characterizing the clinical phenotype of multiple-case families, (iii) identifying the underlying genetic mechanism for TGCT susceptibility in families, (iv) evaluating counseling, psychosocial, and behavioral issues resulting from membership in an FTGCT family, and (v) creating an annotated biospecimen repository to permit subsequent translational research studies. Noteworthy findings include (i) documenting the epidemiologic similarities between familial and sporadic TGCT, (ii) demonstrating significantly younger age-at-diagnosis for familial vs. sporadic TGCT, (iii) absence of a dysmorphic phenotype in affected family members, (iv) shifting the focus of gene discovery from a search for rare, highly penetrant susceptibility variants to the hypothesis that multiple, more common, lower penetrance genes underlie TGCT genetic risk, (v) implicating testicular microlithiasis in FTGCT risk, and (vi) observing that aberrant methylation may contribute to FTGCT risk. A clinically based, biospecimen-intensive, multidisciplinary research strategy has provided novel, valuable insights into the etiology of FTGCT, and created a research resource which will support FTGCT clinical and laboratory studies for years to come.

Maximum sustainable speed, energetics and swimming kinematics of a tropical carangid fish, the green jack Caranx caballus.

Maximum sustained swimming speeds, swimming energetics and swimming kinematics were measured in the green jack Caranx caballus (Teleostei: Carangidae) using a 41 l temperature-controlled, Brett-type swimming-tunnel respirometer. In individual C. caballus [mean ±s.d. of 22·1 ± 2·2 cm fork length (L(F) ), 190 ± 61 g, n = 11] at 27·2 ± 0·7° C, mean critical speed (U(crit)) was 102·5 ± 13·7 cm s⁻¹ or 4·6 ± 0·9 L(F) s⁻¹. The maximum speed that was maintained for a 30 min period while swimming steadily using the slow, oxidative locomotor muscle (U(max,c)) was 99·4 ± 14·4 cm s⁻¹ or 4·5 ± 0·9 L(F) s⁻¹. Oxygen consumption rate (M in mg O2 min⁻¹) increased with swimming speed and with fish mass, but mass-specific M (mg O2 kg⁻¹ h⁻¹) as a function of relative speed (L(F) s⁻¹) did not vary significantly with fish size. Mean standard metabolic rate (R(S) ) was 170 ± 38 mg O2 kg⁻¹ h⁻¹, and the mean ratio of M at U(max,c) to R(S) , an estimate of factorial aerobic scope, was 3·6 ± 1·0. The optimal speed (U(opt) ), at which the gross cost of transport was a minimum of 2·14 J kg⁻¹ m⁻¹, was 3·8 L(F) s⁻¹. In a subset of the fish studied (19·7-22·7 cm L(F) , 106-164 g, n = 5), the swimming kinematic variables of tailbeat frequency, yaw and stride length all increased significantly with swimming speed but not fish size, whereas tailbeat amplitude varied significantly with speed, fish mass and L(F) . The mean propulsive wavelength was 86·7 ± 5·6 %L(F) or 73·7 ± 5·2 %L(T) . Mean ±s.d. yaw and tailbeat amplitude values, calculated from lateral displacement of each intervertebral joint during a complete tailbeat cycle in three C. caballus (19·7, 21·6 and 22·7 cm L(F) ; 23·4, 25·3 and 26·4 cm L(T) ), were 4·6 ± 0·1 and 17·1 ± 2·2 %L(T) , respectively. Overall, the sustained swimming performance, energetics, kinematics, lateral displacement and intervertebral bending angles measured in C. caballus were similar to those of other active ectothermic fishes that have been studied, and C. caballus was more similar to the chub mackerel Scomber japonicus than to the kawakawa tuna Euthynnus affinis.

Composição do óleo essencial de quatro espécies do gênero Plectranthus / Essential oil composition of four Plectranthus species

O gênero Plectranthus é considerado um dos mais ricos em óleos essenciais dentro da família Lamiaceae, compreendendo muitas espécies com propriedades medicinais. Algumas destas são conhecidas popularmente como boldo, as quais possuem semelhanças taxonômicas e diversas sinonímias, possuindo ações anti-dispépticas, analgésicas e digestivas. O objetivo deste estudo foi avaliar quantitativa e qualitativamente os óleos essenciais presentes nas folhas das espécies P. amboinicus, P. barbatus, P. grandis e P. neochilus. A extração do óleo foi realizada por hidrodestilação, utilizando pentano como solvente extrator, repetida por três vezes para cada uma das espécies. A análise dos componentes dos óleos essenciais das quatro espécies de Plectranthus, através da CG/EM, permitiu identificar 14 componentes químicos, sendo a maioria sesquiterpenos. O trans-cariofileno se apresentou em elevada concentração nos óleos estudados. Alguns componentes químicos demonstraram ser específicos para cada espécie e outros apresentaram ocorrência comum a todas as quatro, possibilitando a diferenciação das mesmas em dois grupos, um formado por P. amboinicus e P. neochilus e o outro por P. grandis e P. barbatus. Conclui-se que as quatro espécies de boldo apresentam diferenças significativas quanto ao teor e à constituição química do óleo essencial.

Plectranthus has been considered one of the richest genera in essential oils within the Lamiaceae family, which includes several species with medicinal properties. Some of them are commonly known as boldo and present taxonomic similarities and several synonymies, with antidyspeptic, analgesic and digestive actions. The aim of this study was to evaluate quantitatively and qualitatively the essential oils from P. amboinicus, P. barbatus, P. grandis and P. neochilus leaves. The oil was extracted by hydrodistillation using pentane as extracting solvent and was repeated three times for each species. The analysis of essential oil components by GC/MS in the four Plectranthus species identified 14 chemical components, mostly sesquiterpenes. High concentration of trans-caryophyllene was found in the studied oils. Some chemical components were specific for each species and other components had common occurrence in all four species, allowing their differentiation into two groups, one composed of P. amboinicus and P. neochilus and another one by P. grandis and P. barbatus. In conclusion, the four boldo species had significant differences as to essential oil yield and chemical composition.

Diversidade genética entre acessos de espinheira-santa (Maytenus ilicifolia Mart. ex Reis.) coletados no estado do Rio Grande do Sul, Brasil / Genetic diversity of "espinheira-santa" (Maytenus ilicifolia Mart. ex Reis.) accessions collected in Rio Grande do Sul State, Brazil

Maytenus ilicifolia Mart. ex Reis., popularmente conhecida como espinheira-santa, é espécie autóctone pertencente à família Celastraceae, usada para tratamento de úlceras gástricas e gastrites. Devido à importância medicinal, houve aumento no extrativismo das populações naturais, tornando-a uma espécie prioritária para a conservação, a fim de evitar a erosão genética. Buscou-se com este trabalho analisar a diversidade genética de 20 acessos de M. ilicifolia coletados em diferentes localidades no Rio Grande do Sul. Utilizando marcadores moleculares do tipo AFLP, foram testadas oito combinações de primers, que geraram 455 bandas eletroforéticas, 100 por cento polimórficas. As combinações de primers E-ACC/M-CAA, E-ACG/M-CTA, E-ACG/M-CTC apresentaram o maior número de bandas eletroforéticas, 71 cada, totalizando 46,80 por cento do polimorfismo total. Os valores de similaridade genética calculada pelo coeficiente simple matching foram utilizados para gerar o dendrograma de similaridade pelo método UPGMA. Foi obtido alto coeficiente de correlação cofenética (r=0,94), demonstrando elevada representatividade dos dados de similaridade genética no dendrograma. Pela AMOVA verificou-se que 89,33 por cento da diversidade total ocorreram entre indivíduos dentro das populações. A caracterização molecular de acessos de Maytenus ilicifolia por meio de AFLP foi eficiente para identificar diversidade genética. Através da análise de similaridade genética o banco de germoplasma poderia ser composto com os acessos que apresentaram menor similaridade e maior número de alelos, permitindo com que estes fornecessem ampla cobertura do genoma que compõem Maytenus ilicifolia. Os acessos que ficaram agrupados em mesmo cluster e com número reduzido de alelos podem ser descartados deste banco. A diversidade genética intrapopulacional identificada por esse marcador foi muito maior do que aquela entre populações.

Maytenus ilicifolia Mart. ex Reis., popularly known as "espinheira-santa", is an autochthonous species belonging to the Celastraceae family. This species is used to treat ulcers and gastritis. Due to its medicinal importance, the exploitation of natural populations has increased, making the conservation of this species essential to prevent genetic erosion. The aim of this work was to analyze the genetic diversity of 20 M. ilicifolia accessions collected in different localities of Rio Grande do Sul State. Using AFLP-type molecular markers, eight primer combinations were tested, producing 455 electrophoretic profiles, with 100 percent polymorphism. The primer combinations E-ACC/M-CAA, E-ACG/M-CTA and E-ACG/M-CTC presented the largest number of electrophoretic profiles, 71 each, totaling 46.80 percent of the total polymorphism. The values of genetic similarity estimated by Simple Matching Coefficient were used to produce the dendrogram of similarity by the UPGMA method. A high cophenetic correlation coefficient (r = 0.94) was obtained, demonstrating high representativeness of the data of genetic similarity in the dendrogram. Using AMOVA, 89.33 percent of the total diversity were observed among individuals from the same population. The molecular characterization of Maytenus ilicifolia accessions by AFLP allowed the identification of genetic diversity. The genetic similarity analysis indicated that the germplasm bank could be composed of the accessions presenting the lowest similarity and the largest numbers of alleles, providing a comprehensive coverage of Maytenus ilicifolia genome. The accessions that were grouped into one same cluster and with a reduced number of alleles could be disposed of this bank. The genetic diversity identified by this marker within populations was much greater than that between populations.

Magnetoamplification in a bipolar magnetic junction transistor.

We have demonstrated the first bipolar magnetic junction transistor using a dilute magnetic semiconductor. For an InMnAs p-n-p transistor magnetoamplification is observed at room temperature. The observed magnetoamplification is attributed to the magnetoresistance of the magnetic semiconductor InMnAs heterojunction. The magnetic field dependence of the transistor characteristics confirm that the magnetoamplification results from the junction magnetoresistance. To describe the experimentally observed transistor characteristics, we propose a modified Ebers-Moll model that includes a series magnetoresistance attributed to spin-selective conduction. The capability of magnetic field control of the amplification in an all-semiconductor transistor at room temperature potentially enables the creation of new computer logic architecture where the spin of the carriers is utilized.

Novel structural determinants of single-channel conductance in nicotinic acetylcholine and 5-hydroxytryptamine type-3 receptors.

Nicotinic ACh (acetylcholine) and 5-HT3 (5-hydroxytryptamine type-3) receptors are cation-selective ion channels of the Cys-loop transmitter-gated ion channel superfamily. Numerous lines of evidence indicate that the channel lining domain of such receptors is formed by the alpha-helical M2 domain (second transmembrane domain) contributed by each of five subunits present within the receptor complex. Specific amino acid residues within the M2 domain have accordingly been demonstrated to influence both single-channel conductance (gamma) and ion selectivity. However, it is now clear from work performed on the homomeric 5-HT3A receptor, heteromeric 5-HT3A/5-HT3B receptor and 5-HT3A/5-HT3B receptor subunit chimaeric constructs that an additional major determinant of gamma resides within a cytoplasmic domain of the receptor termed the MA-stretch (membrane-associated stretch). The MA-stretch, within the M3-M4 loop, is not traditionally thought to be implicated in ion permeation and selection. Here, we describe how such observations extend to a representative neuronal nicotinic ACh receptor composed of alpha4 and beta2 subunits and, by inference, probably other members of the Cys-loop family. In addition, we will attempt to interpret our results within the context of a recently developed atomic scale model of the nicotinic ACh receptor of Torpedo marmorata (marbled electric ray).

Histochemistry defines a proteoglycan-rich layer in bovine flexor tendon subjected to bending.

Mid-substance fibrocartilage develops in bovine deep flexor tendon at the point where the tendon wraps under sesamoid bones of the foot and receives transverse compressive loading during locomotion. Fibrocartilage extends several millimeters into the tendon at this location and the proteoglycan-rich tissue stains intensely with Alcian blue. Using histochemical techniques we demonstrate the presence of aggrecan, type VI collagen, and hyaluronic acid in the extracellular matrix of this region of tendon. Biglycan staining was localized to the cells, however. Adjacent to the fibrocartilage, at the outer curvature of the tendon as it bends, the tissue resembles typical tensile tendon with dense bundles of linearly arranged collagen. Longitudinal sections revealed discrete layers of Alcian blue-stained material between the collagen bundles. We demonstrate that these layers of loose matrix also contain aggrecan, type VI collagen, and hyaluronic acid. However, the dense collagen bundles of this region are negative for these components. Transverse sections of tendon in the area adjacent to fibrocartilage show a distinct Alcian blue-stained structure surrounding vascular elements at the point where several fiber bundles come together. This is concluded to be the same structure as the Alcian blue-stained layers seen in longitudinal sections. These observations suggest that proteoglycan-rich matrices in tendon subjected to mechanical loading other than pure tension may serve multiple roles. Such matrices can not only provide compressive stiffness and separate and lubricate collagen bundles that move relative to each other, but may also protect the integrity of vasculature in tendon subjected to bending and shear.

The 5-hydroxytryptamine type 3 (5-HT3) receptor reveals a novel determinant of single-channel conductance.

5-HT3 (5-hydroxytryptamine type 3) receptors are cation-selective ion channels of the Cys-loop transmitter-gated ion channel superfamily. Two 5-HT3 receptor subunits, 5-HT3A and 5-HT3B, have been characterized in detail, although additional putative 5-HT3 subunit genes (HTR3C, HTR3D and HTR3E) have recently been reported. 5-HT3 receptors function as homopentameric assemblies of the 5-HT3 subunit, or heteropentamers of 5-HT3A and 5-HT3B subunits of unknown stoichiometry. The single-channel conductances of human recombinant homomeric and heteromeric 5-HT3 receptors are markedly different, being <1 and approx. 16 pS respectively. Paradoxically, from the results of studies performed on the closely related nicotinic acetylcholine receptor, the channel-lining M2 domain of the 5-HT3A subunit is predicted to enhance cation conduction, whereas that of the 5-HT3B subunit would not. The present study describes a novel determinant of single-channel conductance, out with the M2 domain, which accounts for this anomaly. Utilizing a panel of chimaeric 5-HT3A and 5-HT3B subunits, a profound determinant of single-channel conductance was traced to a putative amphipathic helix (the 'HA stretch') within the large cytoplasmic loop of the receptor. Replacement of three arginine residues (R432, R436 and R440) unique to the HA stretch of the 5-HT3A subunit with the aligned residues (Q395, D399 and A403) of the 5-HT3B subunit increased the single-channel conductance 28-fold. Significantly, from ultrastructural studies of the Torpedo nicotinic acetylcholine receptor, the key residues may be components of narrow openings within the inner vestibule of the channel, located in the cytoplasm, which contribute to the permeation pathway. Our findings indicate an important and hitherto unappreciated function for the HA stretch in the Cys-loop family of transmitter-gated ion channels.

The interaction of anaesthetic steroids with recombinant glycine and GABAA receptors.

BACKGROUND: Anaesthetic steroids are established positive allosteric modulators of GABAA receptors, but little is known concerning steroid modulation of strychnine-sensitive glycine receptors, the principal mediators of fast, inhibitory neurotransmission in the brain stem and spinal cord. This study compared the modulatory actions of five anaesthetic pregnane steroids and two non-anaesthetic isomers at human recombinant alpha1 glycine and alpha1beta2gamma2L GABAA receptors. METHODS: Recombinant alpha1 glycine or alpha1beta2gamma2L GABAA receptors were expressed in Xenopus laevis oocytes and agonist-evoked currents recorded under voltage-clamp. Steroid modulation of currents evoked by GABA, or glycine, was quantified by determining the potency (EC50) and maximal effect of the compounds. RESULTS: The anaesthetics minaxolone (EC50=1.3 microM), Org20599 (EC50=1.1 microM) and alphaxalone (EC50=2.2 microM) enhanced currents mediated by GABAA receptors. The anaesthetics also enhanced currents mediated by glycine receptors, although with higher EC50 values (minaxolone 13.1 microM; Org20599=22.9 microM and alphaxalone=27.8 microM). The maximal enhancement (to 780-950% of control) produced by the three steroids acting at the GABAA receptor was similar, but currents evoked by glycine were potentiated with increasing effectiveness by alphaxalone (199%)

GABAA receptor modulation by the novel intravenous general anaesthetic E-6375.

E-6375 (4-butoxy-2-[4-(2-cyanobenzoyl)-1-piperazinyl] pyrimidine hydrochloride) is a new intravenous general anaesthetic with an anaesthetic potency, in mice, comparable to propofol, or etomidate. Here, we examined the effect of E-6375 upon the GABAA receptor, a putative target of intravenous anaesthetic action. E-6375 reversibly enhanced GABA-evoked currents mediated by recombinant GABAA (alpha1beta2gamma2L) receptors expressed in Xenopus laevis oocytes, with little effect on NMDA- and kainate-evoked currents mediated by NR1a/NR2A and GluR1o/GluR2o glutamate receptors, respectively. E-6375 prolonged the decay of GABA-evoked miniature inhibitory postsynaptic currents recorded from rat Purkinje neurones demonstrating the anaesthetic also enhanced the activity of synaptic GABAA receptors. The GABA enhancing action of E-6375 on recombinant GABAA receptors was unaffected by the subtype of the alpha isoform (i.e. alphaxbeta2gamma2L; x=1-3) within the receptor, but was increased by the omission of the gamma2L subunit. Receptors incorporating beta2, or beta3, subunits were more sensitive to modulation by E-6375 than those containing the beta1 subunit. The selectivity of E-6375 was largely governed by the identity (serine or asparagine) of a single amino acid residue within the second transmembrane domain of the beta-subunit. The various in vivo actions of general anaesthetics may be mediated by GABAA receptor isoforms that have a differential distribution within the CNS. The identification of agents, such as E-6375, that discriminate between GABAA receptor subtypes may augur the development of general anaesthetics with an improved therapeutic profile.

Modulation of native and recombinant GABA(A) receptors by endogenous and synthetic neuroactive steroids.

Upon administration, certain pregnane steroids produce clear behavioural effects including, anxiolysis, sedation, analgesia, anaesthesia and are anti-convulsant. This behavioural profile is characteristic of compounds that act to enhance the actions of GABA acting at the GABA(A) receptor. In agreement, numerous studies have now demonstrated these steroids to be potent, positive allosteric modulators of the GABA(A) receptor. The pregnane steroids are synthesized in the periphery by endocrine glands such as the adrenals and the ovaries, but are also made by neurons and glial cells in the central nervous system itself. Hence, these compounds could play both an endocrine and a paracrine role to influence neuronal excitability by promoting inhibition. Here we review evidence that the pregnane steroids are highly selective and extremely potent GABA(A) receptor modulators and that their effects at 'physiological' concentrations (low nanomolar) may be influenced by the subunit composition of the GABA(A) receptor. This feature may underlie recent findings demonstrating the effects of the neurosteroids on inhibitory synaptic transmission to be brain region dependent, although recent reports suggest that phosphorylation mechanisms may additionally influence neurosteroid sensitivity of the GABA(A) receptor. Numerous synthetic steroids have been synthesized in an attempt to therapeutically exploit the behavioural effects of the pregnane steroids and progress with this approach will be discussed. However, the demonstration that the steroids may be made within the central nervous system offers the alternative strategy of targeting the enzymes that synthesize/metabolise the neurosteroids to exploit this novel endocrine/paracrine interaction.

Neurosteroid modulation of recombinant and synaptic GABAA receptors.

Certain pregnane steroids are now established as potent, positive allosteric modulators of the gamma-aminobutyric acid type A (GABAA) receptor. These compounds are known to be synthesized in the periphery by endocrine glands, such as the ovaries and the adrenal glands, and can rapidly cross the blood-brain barrier. Therefore, such steroids could act as endogeneous modulators of the major inhibitory receptor in the mammalian central nervous system. However, the demonstration that certain neurons and glia can synthesize the pregnane steroids (i.e., neurosteroids) additionally suggests that they may serve a paracrine role by influencing GABAA-receptor function through their local release in the brain itself. Here, we demonstrate that these neurosteroids are highly selective and extremely potent modulators of the GABAA receptor. The subunit composition of the GABAA receptor may influence the actions of the neurosteroids, particularly when considering concentrations of these agents thought to occur physiologically, which may underlie their reported differential effects at certain inhibitory synapses. However, recent work suggests that the phosphorylation status of either the synaptic GABAA receptor or its associated proteins may also influence neurosteroid sensitivity; these findings are discussed. Upon administration, the neurosteroids exhibit clear behavioral effects, including sedation, anticonvulsant actions, and behaviors predictive of anxiolysis; when given at high doses, they induce general anesthesia. Numerous synthetic steroids have been synthesized in an attempt to therapeutically exploit these properties, and these data are reviewed in this chapter. However, targeting the brain enzymes that synthesize and metabolize the neurosteroids may offer a new approach to exploit this novel endocrine-paracrine neurotransmitter interaction.

Alpha-amino acid phenolic ester derivatives: novel water-soluble general anesthetic agents which allosterically modulate GABA(A) receptors.

In the search for a novel water-soluble general anesthetic agent the activity of an alpha-amino acid phenolic ester lead, identified from patent literature, was markedly improved. In addition to improving in vivo activity in mice, good in vitro activity at GABA(A) receptors was also conferred. Within the series of compounds good enantioselectivity for both in vitro and in vivo activity was found, supporting a protein-mediated mechanism of action for anesthesia involving allosteric modulation of GABA(A) receptors. alpha-Amino acid phenolic ester 19, as the hydrobromide salt Org 25435, was selected for clinical evaluation since it retained the best overall anesthetic profile coupled with improved stability and water solubility. In the clinic it proved to be an effective intravenous anesthetic in man with rapid onset of and recovery from anesthesia at doses of 3 and 4 mg/kg.