Serological identification of house dust mite allergens in dogs with atopic dermatitis

House dust mite antigens have been used for decades to diagnose allergic diseases in humans and animals. The objective of this study was to identify allergens in commercial Dermatophagoides farinae and Blomia tropicalis extracts by immunoblotting using sera from allergic dogs and anti-dog IgE conjugate. The analysis of antigens present in the D. farinae extract (FDA Allergenic) using sera from 10 dogs allergic to D. farinae showed that eight sera recognized a band of approximately 102 kDa, eight recognized two bands of 52 to 76 kDa, five recognized one band of approximately 76 kDa, four recognized one band of 31 to 38 kDa, and two recognized one band of 12 to 17 kDa. Immunoblot assays of the B. tropicalis extract (FDA Allergenic) using sera from 10 animals allergic to B. tropicalis showed that five sera recognized two bands of 52 to 76 kDa. These results demonstrate the importance of the two house dust mite species for the pathogenesis of canine atopic dermatitis in Brazil. In addition, the results indicate which allergens should be present in allergenic extracts used for diagnosis and allergen-specific immunotherapy.

In-House Validation of Capillary Electrophoresis Method

Capillary electrophoresis method designed originally for the analysis of monosaccharides was validated using reference solutions of polydatin. The validation was conducted by studying and determining the concentration levels of LOD and LOQ and the range of linearity and by determining levels of uncertainty in respect to repeatability and reproducibility. The reliability of the gained results is also discussed. A guide with recommendations considering the validation and overall design of analysis sequences with CE is also produced as a result of this study.

Relative influence of age, resting heart rate and sedentary life style in short-term analysis of heart rate variability

In order to assess the relative influence of age, resting heart rate (HR) and sedentary life style, heart rate variability (HRV) was studied in two different groups. The young group (YG) consisted of 9 sedentary subjects aged 15 to 20 years (YG-S) and of 9 nonsedentary volunteers (YG-NS) also aged 15 to 20. The elderly sedentary group (ESG) consisted of 16 sedentary subjects aged 39 to 82 years. HRV was assessed using a short-term procedure (5 min). R-R variability was calculated in the time-domain by means of the root mean square successive differences. Frequency-domain HRV was evaluated by power spectrum analysis considering high frequency and low frequency bands. In the YG the effort tolerance was ranked in a bicycle stress test. HR was similar for both groups while ESG showed a reduced HRV compared with YG. Within each group, HRV displayed a negative correlation with HR. Although YG-NS had better effort tolerance than YG-S, their HR and HRV were not significantly different. We conclude that HRV is reduced with increasing HR or age, regardless of life style. The results obtained in our short-term study agree with others of longer duration by showing that age and HR are the main determinants of HRV. Our results do not support the idea that changes in HRV are related to regular physical activity.

Cloning, sequence analysis, and expression of cDNA coding for the major house dust mite allergen, Der f 1, in Escherichia coli

Our objective was to clone, express and characterize adult Dermatophagoides farinae group 1 (Der f 1) allergens to further produce recombinant allergens for future clinical applications in order to eliminate side reactions from crude extracts of mites. Based on GenBank data, we designed primers and amplified the cDNA fragment coding for Der f 1 by nested-PCR. After purification and recovery, the cDNA fragment was cloned into the pMD19-T vector. The fragment was then sequenced, subcloned into the plasmid pET28a(+), expressed in Escherichia coli BL21 and identified by Western blotting. The cDNA coding for Der f 1 was cloned, sequenced and expressed successfully. Sequence analysis showed the presence of an open reading frame containing 966 bp that encodes a protein of 321 amino acids. Interestingly, homology analysis showed that the Der p 1 shared more than 87% identity in amino acid sequence with Eur m 1 but only 80% with Der f 1. Furthermore, phylogenetic analyses suggested that D. pteronyssinus was evolutionarily closer to Euroglyphus maynei than to D. farinae, even though D. pteronyssinus and D. farinae belong to the same Dermatophagoides genus. A total of three cysteine peptidase active sites were found in the predicted amino acid sequence, including 127-138 (QGGCGSCWAFSG), 267-277 (NYHAVNIVGYG) and 284-303 (YWIVRNSWDTTWGDSGYGYF). Moreover, secondary structure analysis revealed that Der f 1 contained an a helix (33.96%), an extended strand (17.13%), a ß turn (5.61%), and a random coil (43.30%). A simple three-dimensional model of this protein was constructed using a Swiss-model server. The cDNA coding for Der f 1 was cloned, sequenced and expressed successfully. Alignment and phylogenetic analysis suggests that D. pteronyssinus is evolutionarily more similar to E. maynei than to D. farinae.

The house that the Court built. The retained powers case law of the European Court of Justice and the constitutional structure of the Union

Euroopan unionin perustamissopimusten katsotaan muodostavan EU:n valtiosäännön, jonka rajat ovat kuitenkin perustamissopimusten määräysten väljän muotoilun sekä Euroopan unionin tuomioistuimen tulkintakäytännön valossa epätarkat. Etenkin kysymys unionin ja sen jäsenvaltioiden välisestä toimivallanjaosta on EU-oikeudellisen tutkimuksen klassikoita. Tarkastelen pro gradu -tutkielmassani unionin valtiosääntörakennetta ja unionioikeuden kokonaisvaikutusta jäsenvaltioiden toimivaltojen käyttöön erityisesti EU-tuomioistuimen ratkaisukäytännössään kehittämän niin kutsutun retained powers -doktriinin valossa. Kyseisen opin mukaan EU-oikeus asettaa vaatimuksia jäsenvaltioiden toimivaltojen käytölle myös niillä aloilla, joilla sääntelytoimivalta on jäänyt jäsenvaltioille eikä sitä ole jaettu unionin kanssa. Aiheen teoreettisen tarkastelun pohjalta analysoin Euroopaun unionin tuomioistuimen ratkaisukäytäntöä erityisesti yhtä säilytetyn toimivallan alaa, koulutusta, koskevissa tapauksissa pyrkien havaitsemaan typologioita tuomioistuimen ratkaisutoiminnassa. Tutkimus noudattaa EU-valtiosääntöoikeuden metodologiaa. Keskeisenä lähdemateriaalina on siten käytetty unionituomioistuimen ratkaisukäytäntöä, joka heijastaa perustamissopimuksia tarkemmin unionin valtiosääntörakennetta. Oikeuskäytännön analyysi ja tulkinta on suoritettu peilaten sitä vasten unionin kehitystä markkinaorientoituneesta organisaatiosta yleismaailmalliseksi poliittiseksi unioniksi. Tutkielmani loppupäätelmä on, että jäsenvaltiot ovat tietyissä rajoissa hyväksyneet unionituomioistuimen kehittämän doktriinin, ja unionituomioistuin on siten saanut aikaan tosiasiallisen muutoksen EU:n valtiosääntörakenteessa. Retained powers -doktriini on omiaan syventämään eurooppalaista integraatiota ja nostaa kysymyksiä toimivallanjaon merkityksestä unionioikeudessa, perustamissopimusten kyvystä heijastaa unionin valtiosääntörakennetta sekä unionituomioistuimen toiminnan poliittisesta luonteesta.

In-house method validation and occurrence of alpha-, beta-endosulfan, endosulfan sulphate, lambda-cyhalothrin, procymidone and trifluralin residues in strawberry

A method for determination of organohalogen pesticides in strawberry by gas chromatography with electron capture detection was validated and applied in a monitoring program. Linearity, matrix effects, and day effect were evaluated for the analytes alpha-endosulfan, beta-endosulfan, endosulfan sulphate, lambda-cyhalothrin, procymidone, and trifluralin. The linear range varied according to the chromatographic response of the analyte. Significant matrix effects were observed. The mean recoveries ranged from 74.6 to 115.4%, with repeatability standard deviations between 1.6 and 21.0% and intermediate precision between 5.9 and 21.0%. Detection, quantification and decision limit, and detection capacity ranged from 0.003 to 0.007 mg/kg, 0.005 to 0.013 mg/kg; 0.003 to 3.128 mg/kg; and 0.005 to 3.266 mg/kg, respectively. The method was fit for the purpose of monitoring organohalogen residues in strawberries. Residues of these pesticides were detected in 124 of the 186 samples analyzed between 2009 and 2011 in the state of Minas Gerais. Nine of them did not comply with the current legislation requirements; among them, seven (3.8%) had residues of unauthorized pesticide for the culture of strawberry, one (0.5%) had residues above the maximum residue limit, and another one (0.5%) exhibited both non-conformities.

Los Angeles [le bâtiment du tribunal, County Court House] : [photographie de presse] / [Agence Rol]

Référence bibliographique : Rol, 60150

Mélanges historiques et juridiques : formulaires et protocoles ; Style du Parlement, de Guillaume DU BREUIL ; ordonnances et actes divers du XIVe siècle ; obsèques de Charles VI ; extraits des Registres du Parlement.

Contient : Sciendum des notaires et secrétaires du Roi : « Sciendum est quod v[adia] notarii sunt sex solidi parisienses per diem... » [même texte latin dans les mss franç. 6021, f. 36, et 18368, f. 83 ; texte français daus les mss franç. 18239, f. 1, 18281, f. 399 v, etc.] ; « Cy après s'ensuivent aucunes notes et enseignemens, touchant les offices des notaires et secrétaires du Roy nostre sire » ; « Prothocole » ou formules de suscriptions « de lettres closes », et formules de lettres empruntées aux chancelleries de Charles VI et de Charles VII ; Formulaire de la chancellerie de France, de l'époque de Charles V, en latin : « Lilterarum regiarum quedam diriguntur subditis regni... » ; Style du Parlement, par Guillaume DU BREUIL ; Notes concernant les pairs de France (f. 71 v), — les « éveschez estant en régale » (f. 71 v), — les cas royaux (f. 72), — les « cas dont congnoist la court laye » (f. 72), etc ; Ordonnances concernant le Parlement et la justice : Ordonnances de Philippe VI de Valois, 13 mars 1344/5 (f. 75), et décembre 1344 (f. 85 v), — et de Jean II le Bon, mai 1355 (f. 93), et décembre 1363 (f. 107) ; Instruction pour la conservation des « droiz de souverainetez et de ressort et autres droiz roiaulx » dans la ville et baronnie de Montpellier, 8 mai 1372 [Ordonnances des rois de France, t. V, 1736, pp. 479-480] ; Actes divers de Charles V, juillet 1363 [?] (f. 114), et mars 1371/2 (f. 115), — et de Philippe VI de Valois, 10 juillet 1336 (f. 117) ; Ordonnance de Philippe VI de Valois, concernant le jugement des procès en matière réelle, 1330 [Or donnances des rois de France, t. II, 1729, pp. 63-64] ; « La manière de faire les obsèques du roy de France, et mesmement du roy Charles VI », novembre 1422 : « Le duc de Bedfort, régent du royaulme de France... » [cf. ms. français 1850, f. 141] ; « Secuntur plura arresta extracta a registris Parlamenti... » ; Autre série d'arrêts, d'une main plus récente ; Choses dont il faut souhaiter que « Dieu nous gart »