Rapid identification of mosquito species and age by mass spectrometric analysis.

BACKGROUND: A rapid, accurate method to identify and to age-grade mosquito populations would be a major advance in predicting the risk of pathogen transmission and evaluating the public health impact of vector control interventions. Whilst other spectrometric or transcriptomic methods show promise, current approaches rely on challenging morphological techniques or simple binary classifications that cannot identify the subset of the population old enough to be infectious. In this study, the ability of rapid evaporative ionisation mass spectrometry (REIMS) to identify the species and age of mosquitoes reared in the laboratory and derived from the wild was investigated. RESULTS: The accuracy of REIMS in identifying morphologically identical species of the Anopheles gambiae complex exceeded 97% using principal component/linear discriminant analysis (PC-LDA) and 84% based on random forest analysis. Age separation into 3 different age categories (1 day, 5-6 days, 14-15 days) was achieved with 99% (PC-LDA) and 91% (random forest) accuracy. When tested on wild mosquitoes from the UK, REIMS data could determine the species and age of the specimens with accuracies of 91 and 90% respectively. CONCLUSIONS: The accuracy of REIMS to resolve the species and age of Anopheles mosquitoes is comparable to that achieved by infrared spectroscopy approaches. The processing time and ease of use represent significant advantages over current, dissection-based methods. Importantly, the accuracy was maintained when using wild mosquitoes reared under differing environmental conditions, and when mosquitoes were stored frozen or desiccated. This high throughput approach thus has potential to conduct rapid, real-time monitoring of vector populations, providing entomological evidence of the impact of alternative interventions.

Pathogenic mitochondrial tRNA point mutations: nine novel mutations affirm their importance as a cause of mitochondrial disease.

Mutations in the mitochondrial genome, and in particular the mt-tRNAs, are an important cause of human disease. Accurate classification of the pathogenicity of novel variants is vital to allow accurate genetic counseling for patients and their families. The use of weighted criteria based on functional studies-outlined in a validated pathogenicity scoring system--is therefore invaluable in determining whether novel or rare mt-tRNA variants are pathogenic. Here, we describe the identification of nine novel mt--tRNA variants in nine families, in which the probands presented with a diverse range of clinical phenotypes including mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes, isolated progressive external ophthalmoplegia, epilepsy, deafness and diabetes. Each of the variants identified (m.4289T>C, MT-TI; m.5541C>T, MT-TW; m.5690A>G, MT-TN; m.7451A>T, MT-TS1; m.7554G>A, MT-TD; m.8304G>A, MT-TK; m.12206C>T, MT-TH; m.12317T>C, MT-TL2; m.16023G>A, MT-TP) was present in a different tRNA, with evidence in support of pathogenicity, and where possible, details of mutation transmission documented. Through the application of the pathogenicity scoring system, we have classified six of these variants as "definitely pathogenic" mutations (m.5541C>T, m.5690A>G, m.7451A>T, m.12206C>T, m.12317T>C, and m.16023G>A), whereas the remaining three currently lack sufficient evidence and are therefore classed as 'possibly pathogenic' (m.4289T>C, m.7554G>A, and m.8304G>A).

Central representation of somatic sensations in the parietal operculum (SII) and insula.

Four subjects with small restricted cerebral cortical infarcts have been examined. One had a lesion confined to the parietal operculum (SII), while in the second the SII lesion also encroached on the posterior insula; in the third subject, both banks of the sylvian fissure and the dorsal insula were involved, while in the fourth the lesion involved the upper bank of the sylvian fissure. In all cases, the postcentral gyrus (SI) was intact. Subjects 1 and 2 had mild spontaneous pain, but subjects 3 and 4 had never had spontaneous pain. In the affected areas, none could feel mechanical (skinfold pinch) pain. The 2 subjects with spontaneous pain could not discriminate sharpness (pinprick), but this was unimpaired in the third and fourth subjects. Warmth, cold, and heat pain were impaired in the 2 subjects with spontaneous pain, but not in those without; however warm-cold difference was greater in the affected regions of all subjects. The possibility must nevertheless be considered that the presence of central pain in some way alters the cortical mechanisms for the perception of thermal stimuli. Certainly, as we had earlier observed, spontaneous pain only occurs when there is interference with thermal sensation. Functional MRI (fMRI) studies following thermal stimulation in subjects 1 and 2 showed these areas, particularly SII, to be concerned with the reception of innocuous and noxious thermal stimuli, mechanical (skinfold pinch) pain and sharpness (pinprick), implying that SI is principally concerned with the reception of low-intensity mechanical stimuli, although it was activated in 1 of our fMRI-studied subjects by innocuous cooling.