Whole-genome sequencing for prediction of Mycobacterium tuberculosis drug susceptibility and resistance : a retrospective cohort study

BACKGROUND : Diagnosing drug-resistance remains an obstacle to the elimination of tuberculosis. Phenotypic drugsusceptibility testing is slow and expensive, and commercial genotypic assays screen only common resistancedetermining mutations. We used whole-genome sequencing to characterise common and rare mutations predicting drug resistance, or consistency with susceptibility, for all fi rst-line and second-line drugs for tuberculosis. METHODS : Between Sept 1, 2010, and Dec 1, 2013, we sequenced a training set of 2099 Mycobacterium tuberculosis genomes. For 23 candidate genes identifi ed from the drug-resistance scientifi c literature, we algorithmically characterised genetic mutations as not conferring resistance (benign), resistance determinants, or uncharacterised. We then assessed the ability of these characterisations to predict phenotypic drug-susceptibility testing for an independent validation set of 1552 genomes. We sought mutations under similar selection pressure to those characterised as resistance determinants outside candidate genes to account for residual phenotypic resistance. FINDINGS : We characterised 120 training-set mutations as resistance determining, and 772 as benign. With these mutations, we could predict 89·2% of the validation-set phenotypes with a mean 92·3% sensitivity (95% CI 90·7–93·7) and 98·4% specifi city (98·1–98·7). 10·8% of validation-set phenotypes could not be predicted because uncharacterised mutations were present. With an in-silico comparison, characterised resistance determinants had higher sensitivity than the mutations from three line-probe assays (85·1% vs 81·6%). No additional resistance determinants were identifi ed among mutations under selection pressure in non-candidate genes. INTERPRETATION : A broad catalogue of genetic mutations enable data from whole-genome sequencing to be used clinically to predict drug resistance, drug susceptibility, or to identify drug phenotypes that cannot yet be genetically predicted. This approach could be integrated into routine diagnostic workfl ows, phasing out phenotypic drugsusceptibility testing while reporting drug resistance early.Wellcome Trust, National Institute of Health Research, Medical Research Council, and the European Union.http://www.thelancet.com/infectionhb201

An approach to a cost control and pricing policy for road haulage and warehousing

An approach to a cost control and pricing policy for road haulage and warehousin

Intoxicação experimental de bovinos com toxina botulínica tipo D

Realizou-se uma intoxicação experimental em bovinos, pela administração oral, com diferentes doses de toxina botulínica tipo D. O objetivo foi determinar o tempo de permanência da toxina no sangue circulante de bovinos, pela detecção da toxina no soro mediante bioensaio em camundongos, e de verificar a presença da toxina no fígado, no baço, nos rins e no coração, e no conteúdo ruminal de bovinos que morreram e/ou foram sacrificados. Utilizaram-se 12 bovinos, mestiços, divididos em quatro grupos de três animais cada. Os grupos I, II e III receberam 200DL50/ml, 21.300DL50/ml e 63.200DL50/ml de toxina botulínica, respectivamente, e o grupo IV manteve-se como controle. A toxina foi detectada principalmente no soro dos bovinos pertencentes aos grupos II e III que receberam altas doses do inóculo tóxico, nos quais a toxina permaneceu por um período de um a sete dias após o aparecimento dos primeiros sinais clínicos da doença. A toxina não foi detectada no fígado, no baço, nos rins e no coração, mas o foi no conteúdo ruminal de um bovino do grupo II. A toxina botulínica foi mais facilmente detectada no soro do que nos órgãos dos bovinos, sendo encontrada principalmente quando o animal ingeriu muita toxina, durante a fase inicial da doença e por um período de sete dias.With the purpose of generating data to help the laboratory diagnosis of botulism, a bovine experimental intoxication, by oral administration with different doses of botulinic toxin type D was carried out. The objective was to determine the time that toxin can be detected in the circulating blood of bovines, by a mouse bioassay, and also to detect toxin in organs (liver, spleen, kidneys and heart) and in the ruminal contents of bovines that had died or were sacrificed after intoxication. Twelve crossbred bovines were divided into four groups of three animals each. Groups I, II and III received 200DL50/ml, 21,300DL50/ml and 63,200DL50/ml of botulinic toxin, respectively, and group IV was maintained as control. The toxin was detected mainly in the serum of bovines of groups II e III, that received high doses of the toxic inoculum, for a period of 1 to 7 days after signs of illness had appeared. The toxin was not detected in the liver, spleen, kidneys and heart but was found in the ruminal contents of only one bovine of the group II. It was concluded that the botulinic toxin is more easily detected in the serum than in organs of bovines, and it was found out mainly when the animal swallow high amount of toxin and during the initial stage of the illness (7 days)

The unusual interacting pair of galaxies IC 3481 and IC 3481A: An optical-NIR photometric and spectroscopic analysis

Background: Retinopathy of Prematurity (ROP) is the main cause of preventable blindness in premature babies. Currently, there is a shortage of trained ophthalmologists, which has resulted in an alarming increase in cases of vision loss and related complications. This study's aim was to determine the utility of examinations conducted by non-ophthalmologist physicians to assess posterior pole vessel abnormalities in eyes at risk for ROP. Method: Non-ophthalmologist physicians (pediatrician and neonatologist) were trained to use an indirect ophthalmoscope to view the posterior pole of babies at risk for ROP. Examinations were conducted on both eyes of premature infants born before 35 weeks gestational age (GA) starting at the third week after birth and weekly thereafter. The presence of Plus disease was identified by the non-ophthalmologist and results compared to the clinical examination by a pediatric ophthalmologist experienced in ROP detection and treatment. Chi-square was used for proportions and the Mann Whitney U test for medians. Fagan's nomogram was determined for diagnostic usability. The Kappa index was used to rate inter-observer agreement. Results: Results of 228 examinations performed on 150 premature infants were analyzed to determine the correlation of the non-ophthalmologist findings and the eye examination. For any vascular change in posterior pole diagnostic, findings were 87% and 87% accuracy for pediatrician and neonatologist, 82% and 83% sensitivity, 90% and 90% specificity respectively. There was no significant difference found in the detection of Plus disease for the examinations performed by the ophthalmologist compared to those performed by the non-ophthalmologist (P<0.05). Conclusions: After training in the use of an indirect ophthalmoscope, non-ophthalmologist physicians can reliably detect posterior pole retinal vessel changes for ROP diagnosis. " 2011 Informa Healthcare USA, Inc.",,,,,,"10.3109/09286586.2011.602506",,,"http://hdl.handle.net/20.500.12104/45318","http://www.scopus.com/inward/record.url?eid=2-s2.0-80755125555&partnerID=40&md5=4a306744d1c81ec82ce4be19cfdc4c0

The utility of non-ophthalmologist examination of eyes at risk for serious retinopathy of prematurity

Background: Retinopathy of Prematurity (ROP) is the main cause of preventable blindness in premature babies. Currently, there is a shortage of trained ophthalmologists, which has resulted in an alarming increase in cases of vision loss and related complications. This study's aim was to determine the utility of examinations conducted by non-ophthalmologist physicians to assess posterior pole vessel abnormalities in eyes at risk for ROP. Method: Non-ophthalmologist physicians (pediatrician and neonatologist) were trained to use an indirect ophthalmoscope to view the posterior pole of babies at risk for ROP. Examinations were conducted on both eyes of premature infants born before 35 weeks gestational age (GA) starting at the third week after birth and weekly thereafter. The presence of Plus disease was identified by the non-ophthalmologist and results compared to the clinical examination by a pediatric ophthalmologist experienced in ROP detection and treatment. Chi-square was used for proportions and the Mann Whitney U test for medians. Fagan's nomogram was determined for diagnostic usability. The Kappa index was used to rate inter-observer agreement. Results: Results of 228 examinations performed on 150 premature infants were analyzed to determine the correlation of the non-ophthalmologist findings and the eye examination. For any vascular change in posterior pole diagnostic, findings were 87% and 87% accuracy for pediatrician and neonatologist, 82% and 83% sensitivity, 90% and 90% specificity respectively. There was no significant difference found in the detection of Plus disease for the examinations performed by the ophthalmologist compared to those performed by the non-ophthalmologist (P<0.05). Conclusions: After training in the use of an indirect ophthalmoscope, non-ophthalmologist physicians can reliably detect posterior pole retinal vessel changes for ROP diagnosis. © 2011 Informa Healthcare USA, Inc