Use of nonradioactive labeling to detect large gene rearrangements in 21-hydroxylase deficiency

PURPOSE: To establish the Southern blotting technique using hybridization with a nonradioactive probe to detect large rearrangements of CYP21A2 in a Brazilian cohort with congenital adrenal hyperplasia due to 21-hydroxylase deficiency (CAH-21OH). METHOD: We studied 42 patients, 2 of them related, comprising 80 non-related alleles. DNA samples were obtained from peripheral blood, digested by restriction enzyme Taq I, submitted to Southern blotting and hybridized with biotin-labeled probes. RESULTS: This method was shown to be reliable with results similar to the radioactive-labeling method. We found CYP21A2 deletion (2.5%), large gene conversion (8.8%), CYP21AP deletion (3.8%), and CYP21A1P duplication (6.3%). These frequencies were similar to those found in our previous study in which a large number of cases were studied. Good hybridization patterns were achieved with a smaller amount of DNA (5 mug), and fragment signs were observed after 5 minutes to 1 hour of exposure. CONCLUSIONS: We established a non-radioactive (biotin) Southern blot/hybridization methodology for CYP21A2 large rearrangements with good results. Despite being more arduous, this technique is faster, requires a smaller amount of DNA, and most importantly, avoids problems with the use of radioactivity.

Doppler Echocardiographic Assessment of Pregnant Women with Chronic Arterial Hypertension

OBJECTIVE: To assess structural and functional cardiac changes in asymptomatic pregnant women with chronic arterial hypertension (CAH). METHODS: One hundred pregnant women with CAH underwent conventional Doppler echocardiography. The Student t test was used to compare them with 29 normotensive pregnant women (NT) in their third gestational trimester. RESULTS: Systolic (SBP; mmHg) and diastolic (DBP; mmHg) blood pressure values were higher (p<0.001) in the CAH pregnant women (SBP: 139±19 and DBP: 92± 18) as compared with those of the NT group (SBP: 112±10 and DBP: 74±9). A significant enlargement of the left atrium (4.10±0.48 cm vs 3.6±0.3 cm; p<0.001) and of the left ventricular normalized mass (59.6±19.7 g/cm2,7 vs 41.9±3.4 g/cm2,7; p<0.001) was observed. Cardiac output (CO, L/min) and systolic volume (SV, mL) were significantly higher in the CAH group (CO: 6.0±1.54 vs 4.9±2.1, p<0.01; SV: 77.3±19.8 vs 56.5±25.8, p<0.001). CONCLUSION: Chronic hypertensive pregnant women have structural and functional cardiac changes that justify routine cardiologic assessment, even in the absence of cardiopulmonary symptoms.

Opuscules entomologiques / par E. Mulsant.

cah.3e, 1853

Calciuria and preeclampsia

Urinary calcium excretion has been reported to be diminished in preeclampsia. The objective of the present study was to determine urinary calcium excretion in pregnant patients with chronic arterial hypertension (CAH) and preeclampsia (PE), and in normotensive patients (N). Forty-four pregnant patients (gestional age, 20-42 weeks; 18 CAH, 17 PE, 9 N) were evaluated for calciuria, proteinuria, plasma uric acid and blood pressure. Patients with PE (82 ± 15.1 mg/24 h) showed significantly lower calciuria (P<0.05) than the group with CAH (147 ± 24.9 mg/24 h) and the N group (317 ± 86.0 mg/24 h) (P<0.05, Student t-test). Plasma uric acid was significantly higher in the PE group (6.1 ± 0.38 mg/dl) than the CAH group (5.0 ± 0.33 mg/dl; P<0.05), which also presented higher proteinuria levels, although the difference was not statistically significant. Diastolic and systolic blood pressure did not differ between the PE (164 ± 105 mmHg) and CAH (164 ± 107 mmHg) groups. Calciuria was significantly lower in the group with preeclampsia than in the group with chronic arterial hypertension. We conclude that calciuria can be a further factor for identifying preeclampsia

Phenotype and genotype correlation of the microconversion from the CYP21A1P to the CYP21A2 gene in congenital adrenal hyperplasia

Deficiency of 21-hydroxylase is the most common form of congenital adrenal hyperplasia (CAH-21OH). We determined by allele-specific PCR the frequency of microconversion in the CYP21A2 gene in 50 Brazilian patients with the classical (salt wasting: SW and simple virilizing: SV) forms and nonclassical (NC) form of CAH-21OH and correlated genotype with phenotype. Genotypes were classified into three mutation groups (A, B, and C) based on the amount of enzymatic activity in in vitro studies using adrenal cells. In 94 unrelated alleles, we diagnosed 76% of the affected alleles after screening for 7 microconversions. The most frequent point mutations observed in this series were I172N (19%), V281L (18%), and IVS2,A/C>G,-12 (15%). In the SW form, the most frequent mutation was IVS2,A/C>G,-12 (38%), in the SV form it was I172N (53%), and in the NC form it was V281L (57.7%). We observed a good correlation between genotype and phenotype. Discordance between genotype and phenotype was found in one SV patient with a mild mutation in one of the alleles (R356W/V281L). However, we cannot rule out the presence of an additional mutation in these alleles. We also observed a good correlation of genotype with 17alpha-hydroxyprogesterone, testosterone, and androstenedione levels. The severity of external genitalia virilization correlated with the severity of mutation. In conclusion, the frequencies described in the present study did not differ from worldwide studies, including the Brazilian population. The few differences observed may reflect individual sample variations. This new Brazilian cohort study suggests the presence of new mutations in Brazilian patients with different forms of CAH-21OH.

Mélanges littéraires, ou Journal des dames, dédié à la Reine / par M. Dorat

Etat de collection : T. 1, cah. 2 (1777, avr.) ; t. 2, cah. 1 (1777, juin) ; t. 3 (1777, sept.-nov.) ; t. 5-6 (1778, mars-juin)

Análise do mercado de TV por assinatura

A presente tese busca analisar as relações entre as estruturas de produto e custo e a verticalização mediante a aplicação da teoria microeconômica básica. Verificamos que os pacotes de canais ofertados ao consumidor podem ser explicados através de 3 efeitos: Discriminação de Preço, Complementariedade entre grupos de canais e Dispersão da propensão a pagar entre grupos de canais. Dentro das hipóteses indicadas acima, identificamos que a renda do consumidor, o estímulo ao consumo de um canal avançado mediante o consumo de um canal básico e/ou a dispersão da propensão a pagar justificam a estratégia de oferta mediante pacotes de canais de televisão. Constatamos que o mercado de TV por Assinatura é caracterizado pelo elevado poder de mercado associado à infra-estrutura de distribuição (custo fixo alto), e que a verticalização entre as empresas da cadeia produtiva do mercado audiovisual é um fenômeno comum desse mercado. Identificamos, ainda, que é possível existir práticas discriminatórias mediante imposição de limites ao uso de redes de distribuidora por programadoras rivais. Entretanto, a Integração Vertical pode ser motivada por questões de economias tecnológicas ou ganhos de integração de monopólios sucessivos que levam a ganhos de eficiência econômica no mercado e não configuram, portanto, práticas discriminatórias. Ao analisarmos os principais modelos para análise da estrutura de mercado, identificamos que o papel do custo fixo é determinante para a estrutura do mesmo e o número de firmas no mercado não determina a eficiência.

Comparison of four trap types for ambrosia beetles (Coleoptera, Scolytidae) in Brazilian Eucalyptus stands

Eucalyptus spp. plantations represent >60% of the reforested area in Brazil. Although ambrosia beetle attacks on live trees were at first nonexistent, they have begun to appear with greater frequency. Monitoring for pest insects is a key factor in integrated pest management, and baited traps are one of the most widely used methods for insect population detection and survey. We compared the efficiency of the most widely used trap in Brazil to survey for ambrosia beetles and other Scolytidae, the ESALQ-84 type, with other traditionally employed traps: the multiple funnel (Lindgren trap); drainpipe; and slot (Theyson) traps, in a Eucalyptus grandis Hill ex Maiden stand in Brazil. The ESALQ-84 trap was the most efficient in trapping Hypothenemus eruditus Westwood and Hypothenemus obscurus (F.); the multiple funnel trap caught significantly more Cryptocarenus diadematus Eggers; whereas the slot trap caught more Premnobius cavipennis Eichhoff and Xyleborus affinis Eichhoff than the other traps. The drainpipe trap was the least effective trap overall. When corrected for number of beetles caught per trap surface area, catches were significantly higher on the ESALQ-84 trap for the majority of the species analyzed, probably because of a smaller trap surface area. The slot trap was recommended for it caught overall more beetles of the three most economically important scolytid species in eucalypt plantations in Brazil, P. cavipennis, X. affinis, and X, ferrugineus.

Ambrosia and bark beetles (Scolytidae : Coleoptera) in pine and eucalypt stands in southern Brazil

More than 95% of the reforested area in Brazil is covered by exotic Eucalyptus and Pinus plantations. Native Scolytidae, mostly ambrosia beetles, appear to be rapidly adapting to these exotic trees, and reports of economic damage are becoming frequent. The objectives of our research were to survey, characterize and compare the Scolytidae fauna present in a P. taeda and an E. grandis stand in Telemaco Borba, Parana state, Brazil. Beetles were caught in ethanol baited ESALQ-84 vane traps in weekly collections from July 1995 until July 1997. In all, 87 species were trapped, 62 in the pine and 75 in the eucalypt stand. The most abundant beetle species in the pines were Hypothenemus eruditus, Xyleborinus gracilis, Cryptocarenus sp. and Xylosandrus retusus, while the most frequent were H. eruditus, Cryptocarenus sp., H. obscurus, Ambrosiodmus obliquus, and X. gracilis. In the eucalypt stand, H. eruditus, X. retusus, H. obscurus, X. ferrugineus and Microcorthylus minimus were the most abundant species, and H. eruditus, H. obscurus and M. minimus were the most frequently trapped. The majority of the species, regardless of the forest community, were most active between August (end of winter) and October (mid-spring). Significantly more H. eruditus, X. gracilis, Cryptocarenus sp., Corthylus obliquus, Hypothenemus bolivianus, A. obliquus, Sampsonius dampfi and Xyleborus affinis were trapped in the pine stand, while X. retusus, H. obscurus, X. ferrugineus, Xyleborinus linearicollis, Corthylus sp, and Corthylus convexicauda were caught in higher numbers in the eucalypt stand. Approximately 50% of the species trapped were found in both communities. Morisita's similarity index indicates the composition of the two communities is very similar, suggesting that most of the beetles are polyphagous. (C) 2001 Elsevier B.V. B.V. All rights reserved.

Bark and ambrosia beetle (Coleoptera : Scolytidae) responses to volatiles from aging loblolly pine billets

Many species of bark and ambrosia beetles use host volatiles as cues for breeding site location. In a study where the objectives were to identify the different volatiles released by Pinus taeda L. billets as they age, to determine the arrival sequence of scolytids (Colcoptera: Scolytidae), and to correlate volatile emission by the billets with beetle catches, 25 species of scolytids were trapped. Bark beetles were more attracted to the billets in the beginning of the period. whereas ambrosia beetles arrived later. Among the bark beetles, Dendroctonus terebrans (Olivier) was significantly more attracted during the Ist 3 wk after tree felling, Hylastes tenuis Eichhoff in the Ist 2 wk. Pityophtorus pulicarius (Zimmermann) in weeks 2 and 3, and Ips grandicollis (Eichhoff) was more attracted on weeks 3 and 4. Among the ambrosia beetles, Xyleborinus saxeseni (Ratzeburg) was more attracted to billets during weeks 4-6, whereas Xyleborus pubescens Zimmermann and Xyleborus californicus Wood were more attracted during week 6. The billets showed marked decline in attractiveness to all scolytids after 8 wk. Volatiles collected during the beetle trapping periods included 15 hydrocarbon monoterpenes, 18 oxygenated monoterpenes, 4-allylanisole, and ethanol. The hydrocarbon monoterpenes and 4-allylanisole decreased sharply over time, but oxygenated monoterpenes and ethanol increased up to weeks 4-6, after which they also decreased. Good correlations between certain billet volatiles and catches for some beetle species were obtained, but their biological significance could not be determined.

Doppler Echocardiographic Assessment of Pregnant Women with Chronic Arterial Hypertension

OBJECTIVE: To assess structural and functional cardiac changes in asymptomatic pregnant women with chronic arterial hypertension (CAH). METHODS: One hundred pregnant women with CAH underwent conventional Doppler echocardiography. The Student t test was used to compare them with 29 normotensive pregnant women (NT) in their third gestational trimester. RESULTS: Systolic (SBP; mmHg) and diastolic (DBP; mmHg) blood pressure values were higher (p<0.001) in the CAH pregnant women (SBP: 139±19 and DBP: 92± 18) as compared with those of the NT group (SBP: 112±10 and DBP: 74±9). A significant enlargement of the left atrium (4.10±0.48 cm vs 3.6±0.3 cm; p<0.001) and of the left ventricular normalized mass (59.6±19.7 g/cm2,7 vs 41.9±3.4 g/cm2,7; p<0.001) was observed. Cardiac output (CO, L/min) and systolic volume (SV, mL) were significantly higher in the CAH group (CO: 6.0±1.54 vs 4.9±2.1, p<0.01; SV: 77.3±19.8 vs 56.5±25.8, p<0.001). CONCLUSION: Chronic hypertensive pregnant women have structural and functional cardiac changes that justify routine cardiologic assessment, even in the absence of cardiopulmonary symptoms.

Crystallization and preliminary X-ray diffraction analysis of a myotoxic Lys49-PLA(2) from Bothrops jararacussu venom complexed with p-bromophenacyl bromide

For the first time, a non-catalytic and myotoxic Lys49-PLA(2) (BthTX-I from Bothrops jararacussu venom) has been crystallized with BPB inhibitor. X-ray diffraction data were collected and electron-density calculations showed that the ligand is bound to the His48 residue. BthTX-I with His48 chemically modified by BPB shows strongly reduced myotoxic and cytotoxic activities. This suggests a biological correlation between the modification of His48, which is associated with catalytic activity of PLA(2)s, and other toxicological activities of Lys49-PLA(2)s.

Attraction of ambrosia beetles (Coleoptera : Scolytidae) to different tropical pine species in Brazil

Ambrosia beetles are the predominant Scolytidae in Brazil. Little is known about the attractiveness of exotic conifer tree volatiles to native scolytids. Objectives were to compare the attractiveness of logs with and without bark of Pinus oocarpa Schiede, P. caribaea variety bahamensis Barrett & Golfari, P, car. variety caribaea Barrett & Golfari and P. car. variety hondurensis Barrett & Golfari over time to native scolytids in different pine stands, to compare the relative attractiveness of logs relative to ethanol traps, to determine how long it takes for logs to become attractive to ambrosia beetles and when attraction peaks occur, and to determine if volatiles released by live standing trees would mask volatiles released by logs of the same species. In young stands, Hypothenemus was the predominant insect genus, whereas in older stands Xyleborus predominated. Debarked logs trapped more beetles than logs with bark. Pine log species attractiveness was not influenced by volatiles present in the stand. Beetles were divided into the following 3 groups, based on response to log volatiles and ethanol: (1) species attracted to ethanol and not responding to pine terpenes Ambrosiodmus hagedorni (Iglesia), A. retusus (Eichhoff), X. spinulosus Blandford, Corthylus schaufussi Schiede, Cryptocarenus heveae (Hagedorn), H. obscurus (F.), (2) species attracted to ethanol but responding to pine terpenes Xyleborinus gracilis (Eichhoff), X. affinis Eichhoff, H. eruditus Westwood, Premnobius cavipennis Eichhoff, and (3) species more attracted to pine terpenes and less responsive to ethanol, A. obliquus (Le Conte), X. ferrugineus F,, X. catulus Blandford. Pinus car. ;variety bahamensis was the least attractive pine, P. oocarpa the most attractive. The attraction peak varied according to the season; logs were not attractive to beetles 10 wk after cutting.

Identification of sulcatol, a potential pheromone of the ambrosia beetle Gnathotrichus materiarius (Col., Scolytidae)

We report the identification of a potential pheromone for Gnathotrichus materiarius (Fitch) (Col., Scolytidae). The population sex ratio is close to 1:1, and males initiate attacks on host trees. Headspace and hindgut samples from single males showed the presence of the putative pheromone 6-methyl-5-hepten-2-ol, sulcatol. Unmated males released sulcatol for at least 12 days, and ceased producing the pheromone after 20 days. The peak sulcatol release occurred after 2 days. Males cease production of sulcatol 24 h after being paired with females. Single females were unable to initiate galleries, and no sulcatol was detected from their headspace and hindgut samples. The chiral ratio of the pheromone, observed from headspace samples, was 31% (S)-(+)- and 69% (R)-(-)-sulcatol.

Osseointegration assessment of chairside argon-based nonthermal plasma-treated Ca-P coated dental implants

This study investigated the effect of an Argon-based nonthermal plasma (NTP) surface treatment-operated chairside at atmospheric pressure conditions applied immediately prior to dental implant placement in a canine model. Surfaces investigated comprised: Calcium-Phosphate (CaP) and CaP + NTP (CaP-Plasma). Surface energy was characterized by the Owens-Wendt-Rabel-Kaelble method and chemistry by X-ray photoelectron spectroscopy (XPS). Six adult beagles dogs received 2 plateau-root form implants (n = 1 each surface) in each radii, providing implants that remained 1 and 3 weeks in vivo. Histometric parameters assessed were bone-to-implant contact (BIC) and bone area fraction occupancy (BAFO). Statistical analysis was performed by Kruskall-Wallis (95% level of significance) and Dunn's post-hoc test. The XPS analysis showed peaks of Ca, C, O, and P for the CaP and CaP-Plasma surfaces. Both surfaces presented carbon primarily as hydro-carbon (CAC, CAH) with lower levels of oxidized carbon forms. The CaP surface presented atomic percent values of 38, 42, 11, and 7 for C, O, Ca, and P, respectively, and the CaPPlasma presented increases in O, Ca, and P atomic percent levels at 53, 12, and 13, respectively, in addition to a decrease in C content at 18 atomic percent. At 1 week no difference was found in histometric parameters between groups. At 3 weeks significantly higher BIC and BAFO were observed for CaPPlasma treated surfaces. Surface elemental chemistry was modified by the Ar-based NTP. Ar-based NTP improved bone formation around plateau-root form implants at 3 weeks compared with CaP treatment alone. © 2012 Wiley Periodicals, Inc.