Application of real-time PCR for total airborne bacterial assessment : comparison with epifluorescence microscopy and culture-dependent methods

Traditional culture-dependent methods to quantify and identify airborne microorganisms are limited by factors such as short-duration sampling times and inability to count nonculturableor non-viable bacteria. Consequently, the quantitative assessment of bioaerosols is often underestimated. Use of the real-time quantitative polymerase chain reaction (Q-PCR) to quantify bacteria in environmental samples presents an alternative method, which should overcome this problem. The aim of this study was to evaluate the performance of a real-time Q-PCR assay as a simple and reliable way to quantify the airborne bacterial load within poultry houses and sewage treatment plants, in comparison with epifluorescencemicroscopy and culture-dependent methods. The estimates of bacterial load that we obtained from real-time PCR and epifluorescence methods, are comparable, however, our analysis of sewage treatment plants indicate these methods give values 270-290 fold greater than those obtained by the ''impaction on nutrient agar'' method. The culture-dependent method of air impaction on nutrient agar was also inadequate in poultry houses, as was the impinger-culture method, which gave a bacterial load estimate 32-fold lower than obtained by Q-PCR. Real-time quantitative PCR thus proves to be a reliable, discerning, and simple method that could be used to estimate airborne bacterial load in a broad variety of other environments expected to carry high numbers of airborne bacteria. [Authors]

Pièces relatives à l'histoire de la ville de TULLE. IV

Contient : Extrait de l'inventaire des archives de l'église de Tulle ; Extrait du cartulaire de l'église de Tulle ; Extrait du cartulaire de Saint-Martin de Tulle ; « Libellus apologetico-supplex » pour les Recollets de Tulle, par le P. Victorin Tarneau ; Extrait d'un cartulaire de Saint-Martin de Tulle ; Accord entre les chanoines de Tulle et ceux de Notre-Dame de Rocamadour, pour le règlement des droits de ces derniers (15 mars 1423) ; Extrait des statuts de l'église de Tulle ; Extraits de l'obituaire de l'église de Tulle (Molinier, Obituaires, n° 503 bis) ; Note sur diverses pièces conservées aux archives de l'évêché de Tulle ; Extrait des Decisiones Burdegalenses, de Nicolas Bohier (Lyon, 1579, in-fol.) ; Extrait des registres du Parlement de Bordeaux concernant la ville de Tulle (1555-1581) ; Rôle des noms des présidents et conseillers du Parlement de Bordeaux (décembre 1564) ; original ; Extrait des registres du Parlement de Bordeaux (1555-1563) ; Procès-verbal d'une assemblée du chapitre de Tulle, autorisant l'engagement d'une cloche pour le soulagement des pauvres (23 mai 1691) ; Mandement de Humbert [Ancelin], évêque de Tulle, pour l'usage de la viande et des oeufs durant le carême (16 février 1691) ; Autobiographie de Mascaron ; Lettres de provision de l'évêché de Tulle en faveur de Jules Mascaron (5 janvier 1671) ; Billet mortuaire du même (5 décembre 1703) ; Deux lettres du même, évêque d'Agen (19 février et 10 mars, s. d.) ; Lettre anonyme écrite de Tulle, le 19 avril 1666 ; Lettre de Louis de Guron, évêque de Comminges (8 juin 1688) à A. de Fes ; Lettre de M. de Fes à Baluze (Daux, 16 juin 1688) ; Deux lettres de [Louis de] Guron, évêque-nommé de Tulle, à Mazarin (10 décembre et 23 septembre 1652) ; Lettre de M. de Fes à Baluze (Toulouse, 5 novembre 1681) ; Notes sur les familles Guron et de Rechignevoisin ; Lettre de M. de Fes à Baluze (Toulouse, 18 mars 1688) ; Lettres de Louis de Guron, évêque de Tulle, puis de Comminges ; « Bref de N. S. Père le pape Innocent X, » envoyé à l'évêque de Tulle touchant la question du Jansénisme (21 mars 1654) ; imprimé ; Lettre de Richelieu au cardinal Antoine Barberini (25 juin 1634) ; Liste des événements notables de l'histoire de Tulle de 1545 à 1685 ; « Catalogus abbatum et episcoporum Tutellensium, » par Baluze ; placard imprimé avec nombreuses corrections de la main de l'auteur ; Diplômes de Raoul et de Louis IV pour le monastère de Tulle ; Fragments d'un traité, en latin, sur la vision en Dieu ; Lettres d'Harduin [de Péréfixe], archevêque de Paris, authentiquant des reliques rapportées par Baluze (s. d.) ; Certificat d'authenticité de ces mêmes reliques, délivré par Gaspard, cardinal prêtre du titre de San Silvestro in capite (8 novembre 1681) ; placard imprimé ; Lettres adressées à Baluze ; Copies par Baluze de diverses pièces relatives aux reliques rapportées par lui de Rome (1683-1685) ; Extrait d'un rouleau des morts du monastère d'Obazine ; « Ex epigrammatis Joannis Vulteii Remensis » (Lyon, 1537, in-8°), et extraits divers concernant Pierre du Chastel, évêque de Tulle ; Notes relatives aux Privilegia regularium, du P. Bruno Chassaing (Paris, 1648, in-fol.)

Use of long-term suppressive acyclovir after hematopoietic stem-cell transplantation: impact on herpes simplex virus (HSV) disease and drug-resistant HSV disease.

The effect that long-term use of suppressive acyclovir (ACV) has on both overall herpes simplex virus (HSV) disease and ACV-resistant HSV disease was examined in 3 consecutive cohorts of hematopoietic stem-cell transplant (HCT) recipients (n=2049); cohort 1 received ACV for 30 days after HCT, cohort 2 received it for 1 year after HCT, and cohort 3 received it for an extended period (i.e., >1 year) if the patient's immunosuppression continued after 1 year. The 2-year probability of HSV disease was 31.6% (95% confidence interval [CI], 28.0%-35%) in cohort 1, 3.9% (95% CI, 2.7%-5.2%) in cohort 2, and 0% in cohort 3 (P<.001). ACV-resistant HSV disease developed in 10 patients in cohort 1 (2-year probability, 1.3% [95% CI, 0.8%-2.7%]), in 2 patients in cohort 2 (2-year probability, 0.2% [95% CI, 0%-0.8%]; P=.006), and in 0 patients in cohort 3 (cohort 2 vs. cohort 3, P=.3). Long-term use of suppressive prophylactic ACV appears to prevent the emergence of drug-resistant HSV disease in HCT.

La protection des investissements suisses à l'étranger dans le cadre des accords de promotion et de protection des investissements

Dotée d'une économie traditionnellement ouverte vers l'extérieur, la Suisse dispose de l'un des plus vastes réseaux au monde d'accords bilatéraux de promotion et de protection des investissements (APPI). Instrument dont la faible notoriété est inversement proportionnelle au nombre élevé d'accords concernés et aux montants en jeu, les APPI visent à conférer aux entreprises suisses établies à l'étranger - et vice-versa - une large protection contre les risques non commerciaux. Le présent ouvrage s'attache à l'analyse du développement et du contenu des APPI conclus par la Suisse avec ses partenaires, essentiellement des pays en développement. S'appuyant sur la doctrine et la jurisprudence internationales, il traite de manière détaillée des droits matériels et procéduraux parfois contestés que ce type d'accords offrent aux investisseurs et met en évidence les particularités des APPI du réseau suisse.

Is Less Chemotherapy Detrimental in Adults with Philadelphia Chromosome (Ph)-Positive Acute Lymphoblastic Leukemia (ALL) Treated with High-Dose Imatinib? Results of the Prospective Randomized Graaph-2005 Study

Aim: To compare a less intensive regimen based on high-dose imatinib (IM) to an intensive IM/HyperCVAD regimen in adults with Ph+ ALL, in terms of early response and outcome after stem cell transplantation (SCT). Methods: Patients aged 18-60 years with previously untreated Ph+ ALL not evolving from chronic myeloid leukemia were eligible if no contra-indication to chemotherapy and SCT (ClinicalTrials.gov ID, NCT00327678). After a steroid prephase allowing Ph and/or BCR-ABL diagnosis, cycle 1 differed between randomization arms. In arm A (IM-based), IM was given at 800 mg on day 1-28, combined with vincristine (2 mg, day 1, 8, 15, 22) and dexamethasone (40 mg, day 1-2, 8-9, 15-16, and 22-23) only. In arm B (IM/HyperCVAD), IM was given at 800 mg on day 1-14, combined with adriamycin (50 mg/m2, day 4), cyclophosphamide (300 mg/m2/12h, day 1, 2, 3), vincristine (2 mg, day 4 and 11), and dexamethasone (40 mg, day 1-4 and 11-14). All patients received a cycle 2 combining high-dose methotrexate (1 g/m2, day 1) and AraC (3 g/m2/12h, day 2 and 3) with IM at 800 mg on day 1-14, whatever their response. Four intrathecal infusions were given during this induction/consolidation period. Minimal residual disease (MRD) was centrally evaluated by quantitative RQ-PCR after cycle 1 (MRD1) and cycle 2 (MRD2). Major MRD response was defined as BCR-ABL/ABL ratio <0.1%. Then, all patients were to receive allogeneic SCT using related or unrelated matched donor stem cells or autologous SCT if no donor and a major MRD2 response. IM/chemotherapy maintenance was planned after autologous SCT. In the absence of SCT, patients received alternating cycles 1 (as in arm B) and cycles 2 followed by maintenance, like in the published IM/HyperCVAD regimen. The primary objective was non-inferiority of arm A in term of major MRD2 response. Secondary objectives were CR rate, SCT rate, treatment- and transplant-related mortality, relapse-free (RFS), event-free (EFS) and overall (OS) survival. Results: Among the 270 patients randomized between May 2006 and August 2011, 265 patients were evaluable for this analysis (133 arm A, 132 arm B; median age, 47 years; median follow-up, 40 months). Main patient characteristics were well-balanced between both arms. Due to higher induction mortality in arm B (9 versus 1 deaths; P=0.01), CR rate was higher in the less intensive arm A (98% versus 89% after cycle 1 and 98% versus 91% after cycle 2; P= 0.003 and 0.006, respectively). A total of 213 and 205 patients were evaluated for bone marrow MRD1 and MRD2. The rates of patients reaching major MRD response and undetectable MRD were 45% (44% arm A, 46% arm B; P=0.79) and 10% (in both arms) at MRD1 and 66% (68% arm A, 63.5% arm B; P=0.56) and 25% (28% arm A, 22% arm B; P=0.33) at MRD2, respectively. The non-inferiority primary endpoint was thus demonstrated (P= 0.002). Overall, EFS was estimated at 42% (95% CI, 35-49) and OS at 51% (95% CI, 44-57) at 3 years, with no difference between arm A and B (46% versus 38% and 53% versus 49%; P=0.25 and 0.61, respectively). Of the 251 CR patients, 157 (80 arm A, 77 arm B) and 34 (17 in both arms) received allogeneic and autologous SCT in first CR, respectively. Allogeneic transplant-related mortality was similar in both arms (31.5% versus 22% at 3 years; P=0.51). Of the 157 allografted patients, 133 had MRD2 evaluation and 89 had MRD2 <0.1%. In these patients, MRD2 did not significantly influence post-transplant RFS and OS, either when tested with the 0.1% cutoff or as a continuous log covariate. Of the 34 autografted patients, 31 had MRD2 evaluation and, according to the protocol, 28 had MRD2 <0.1%. When restricting the comparison to patients achieving major MRD2 response and with the current follow-up, a trend for better results was observed after autologous as compared to allogeneic SCT (RFS, 63% versus 49.5% and OS, 69% versus 58% at 3 years; P=0.35 and P=0.08, respectively). Conclusions: In adults, the use of TK inhibitors (TKI) has markedly improved the results of Ph+ ALL therapy, now close to those observed in Ph-negative ALL. We demonstrated here that chemotherapy intensity may be safely reduced when associated with high-dose IM. We will further explore this TKI-based strategy using nilotinib prior to SCT in our next GRAAPH-2013 trial. The trend towards a better outcome after autologous compared to allogeneic SCT observed in MRD responders validates MRD as an important early surrogate endpoint for treatment stratification and new drug investigation in this disease.

Efeito da densidade de plantas e da adubação NPK na produção inicial de palmito de pupunheira

Avaliaram-se os efeitos da densidade de plantas e doses de adubação na produção de palmito de pupunheira, em condições de campo, na Fazenda Yuricam, Rio Preto da Eva (AM). O delineamento experimental foi de blocos casualizados com três repetições, com os tratamentos em arranjo fatorial 3 x 5, sendo avaliados os fatores: densidade de plantas (10.000; 5.000 e 3.333 plantas ha-1) e doses de adubação NPK (0-0-0; 112,5-12,5-90; 337,5-38-270; 225-90-180, 225-25-180 em kg ha-1 de N, P2O5 e K2O, respectivamente). Os resultados do primeiro ano de produção de palmito, em termos de estipes colhidas, para efeito das doses de adubação NPK, foram: 225-90-180 (66 %); 225-25-180 (57 %); 337,5-38-270 (50 %); 112,5-12,5-90 (26 %); 0-0-0 (1 %) - os números entre parênteses indicam os percentuais de plantas coletadas em relação às plantadas - e para efeitos de densidade: 3.333 plantas ha-1 (58 %); 5.000 plantas ha-1 (28 %); 10.000 plantas ha-1 (40 %). O tempo médio para extração de palmito foi menor na densidade de 3.333 plantas ha-1 (18,4 meses) e na dose de adubação NPK de 225-90-180 (18,1 meses). O efeito simples da adubação NPK dentro da densidade de 10.000 plantas ha-1 com a adubação 225-90-180 e 225-25-180 apresentou maior produção de palmito, com 841 e 779 kg ha-1, e de 1.154 e 1.223 kg ha-1 de estipe tenro, respectivamente, nas duas adubações.

Quantification of brain glycogen concentration and turnover through localized 13C NMR of both the C1 and C6 resonances.

We have recently shown that at isotopic steady state (13)C NMR can provide a direct measurement of glycogen concentration changes, but that the turnover of glycogen was not accessible with this protocol. The aim of the present study was to design, implement and apply a novel dual-tracer infusion protocol to simultaneously measure glycogen concentration and turnover. After reaching isotopic steady state for glycogen C1 using [1-(13)C] glucose administration, [1,6-(13)C(2)] glucose was infused such that isotopic steady state was maintained at the C1 position, but the C6 position reflected (13)C label incorporation. To overcome the large chemical shift displacement error between the C1 and C6 resonances of glycogen, we implemented 2D gradient based localization using the Fourier series window approach, in conjunction with time-domain analysis of the resulting FIDs using jMRUI. The glycogen concentration of 5.1 +/- 1.6 mM measured from the C1 position was in excellent agreement with concomitant biochemical determinations. Glycogen turnover measured from the rate of label incorporation into the C6 position of glycogen in the alpha-chloralose anesthetized rat was 0.7 micromol/g/h.

Measures of health and disease in Africa: are current methods giving us useful information about trends in cardiovascular diseases?

An enormous burst of interest in the public health burden from chronic disease in Africa has emerged as a consequence of efforts to estimate global population health. Detailed estimates are now published for Africa as a whole and each country on the continent. These data have formed the basis for warnings about sharp increases in cardiovascular disease (CVD) in the coming decades. In this essay we briefly examine the trajectory of social development on the continent and its consequences for the epidemiology of CVD and potential control strategies. Since full vital registration has only been implemented in segments of South Africa and the island nations of Seychelles and Mauritius - formally part of WHO-AFRO - mortality data are extremely limited. Numerous sample surveys have been conducted but they often lack standardization or objective measures of health status. Trend data are even less informative. However, using the best quality data available, age-standardized trends in CVD are downward, and in the case of stroke, sharply so. While acknowledging that the extremely limited available data cannot be used as the basis for inference to the continent, we raise the concern that general estimates based on imputation to fill in the missing mortality tables may be even more misleading. No immediate remedies to this problem can be identified, however bilateral collaborative efforts to strength local educational institutions and governmental agencies rank as the highest priority for near term development.

Efeito da Gliricidia sepium sobre nutrientes do solo, microclima e produtividade do milho em sistema agroflorestal no Agreste Paraibano

Gliricidia sepium é uma leguminosa arbórea que tem sido utilizada em sistemas em aléias no semi-árido nordestino por apresentar bom desenvolvimento em condições de estresse hídrico. Entretanto, há pouca informação disponível sobre o efeito da introdução dessa espécie nos agroecossistemas da região. No presente estudo, objetivou-se avaliar a influência da distância de plantas de Gliricidia sepium sobre características da cultura do milho e do solo e microclima no Agreste Paraibano. O estudo foi realizado no município de Esperança (PB), em área de 0,5 ha, onde, em 1996, foram plantadas fileiras de G. sepium espaçadas 6 m entre si e com 1 m entre as árvores. Nesta área, em 2002, foram delimitadas quatro parcelas de 6 x 8 m e, em cada parcela, foi estabelecido um transeto perpendicular às fileiras de árvores com três posições de amostragem: (1) nas fileiras de árvores (0 m); (2) a 1 m das fileiras de árvores, e (3) a 3 m de distância das fileiras de árvores. O delineamento experimental utilizado foi em blocos casualizados com quatro repetições. A massa seca de folhedo caído embaixo da fileira de árvores foi de 1.390 kg ha-1 e diminuiu, gradativamente, para 270 kg ha-1 a 3 m de distância das árvores. As concentrações de P, K e matéria orgânica leve (MOL) embaixo das árvores foram maiores do que a 1 e 3 m de distância das fileiras. As médias mensais das temperaturas mínimas do ar e do solo embaixo e a 3 m das árvores foram similares. Entretanto, as médias mensais das temperaturas máximas do solo e do ar foram de 6 e 2 °C mais altas a 3 m das árvores, respectivamente, ao longo do período de estudo. A umidade do solo foi significativamente menor embaixo das árvores do que a 1 e 3 m de distância. O milho produziu mais grãos e palha e acumulou mais nutrientes nas posições mais próximas das fileiras de G. sepium.