Analytical Performance Characteristics of the Cepheid GeneXpert Ebola Assay for the Detection of Ebola Virus

BackgroundThe recently developed Xpert® Ebola Assay is a novel nucleic acid amplification test for simplified detection of Ebola virus (EBOV) in whole blood and buccal swab samples. The assay targets sequences in two EBOV genes, lowering the risk for new variants to escape detection in the test. The objective of this report is to present analytical characteristics of the Xpert® Ebola Assay on whole blood samples.ConclusionIn summary, we found the Xpert® Ebola Assay to have high analytical sensitivity and specificity for the detection of EBOV in whole blood. It offers ease of use, fast turnaround time, and remote monitoring. The test has an efficient viral inactivation protocol, fulfills inclusivity and exclusivity criteria, and has specimen stability characteristics consistent with the need for decentralized testing. The simplicity of the assay should enable testing in a wide variety of laboratory settings, including remote laboratories that are not capable of performing highly complex nucleic acid amplification tests, and during outbreaks where time to detection is critical.     

Recessive TBC1D24 Mutations Are Frequent in Moroccan Non-Syndromic Hearing Loss Pedigrees

Mutations in the TBC1D24 gene are responsible for four neurological presentations: infantile epileptic encephalopathy, infantile myoclonic epilepsy, DOORS (deafness, onychodystrophy, osteodystrophy, mental retardation and seizures) and NSHL (non-syndromic hearing loss). For the latter, two recessive (DFNB86) and one dominant (DFNA65) mutations have so far been identified in consanguineous Pakistani and European/Chinese families, respectively. Here we report the results of a genetic study performed on a large Moroccan cohort of deaf patients that identified three families with compound heterozygote mutations in TBC1D24. Four novel mutations were identified, among which, one c.641G>A (p.Arg214His) was present in the three families, and has a frequency of 2% in control Moroccan population with normal hearing, suggesting that it acts as an hypomorphic variant leading to restricted deafness when combined with another recessive severe mutation. Altogether, our results show that mutations in TBC1D24 gene are a frequent cause (>2%) of NSHL in Morocco, and that due to its possible compound heterozygote recessive transmission, this gene should be further considered and screened in other deaf cohorts.     

Genetic and molecular analysis of the CLDN14 gene in Moroccan family with non-syndromic hearing loss

BACKGROUND:

Hearing loss is the most prevalent human genetic sensorineural defect. Mutations in the CLDN14 gene, encoding the tight junction claudin 14 protein expressed in the inner ear, have been shown to cause non-syndromic recessive hearing loss DFNB29.

AIM:

We describe a Moroccan SF7 family with non-syndromic hearing loss. We performed linkage analysis in this family and sequencing to identify the mutation causing deafness.

MATERIALS AND METHODS:

Genetic linkage analysis, suggested the involvement of CLDN14 and KCNE1 gene in deafness in this family. Mutation screening was performed using direct sequencing of the CLDN14 and KCNE1 coding exon gene.

RESULTS:

Our results show the presence of c.11C>T mutation in the CLDN14 gene. Transmission analysis of this mutation in the family showed that the three affected individuals are homozygous, whereas parents and three healthy individuals are heterozygous. This mutation induces a substitution of threonine to methionine at position 4.

CONCLUSION:

These data show that CLDN14 gene can be i mplicated in the development of hearing loss in SF7 family; however, the pathogenicity of c.11C>T mutation remains to be determined.     

The comparative efficacy of some botanicals for the control of the seed weevil Caryedon serratus Olivier (Col., Bruchidae) in seeds of Acacia seyal Del.

A comparative study on the efficacy of powder and oil extracts from leaves and seed kernels of Azadirachta indica Juss. and Eucalyptus camaldulensis Dehn. against the seed weevil Caryedon serratus Olivier infesting seeds of Acacia seyal Del. was carried out. EO 1, EO 5 and NSKO5% were comparable to BHC 3% in significantly reducing egg laying by C. serratus. With the exception of ELP and NLP, all botanicals significantly reduced egg hatching. Botanicals significantly reduced larval development with the exception of ELP and NLP. EO and NSKO at 3 and 5% were significantly more effective than BHC in reducing larval development by 95–97%. All botanicals had no significant effect on adult emergence which was only affected by BHC. Botanicals caused 4.3–43% mortality of C. serratus adults and the maximum mortality occurred in EO 3% as compared to BHC. It is apparent that oils extracted from E. camaldulensis had a fumigant and knock down effect on adult C. serratus. Extracted oils were more effective than leaf or seed powder. Oils extracted from leaves of E. camaldulensis were volatile and had a knock down effect on adult C. serratus indicating the possibility of using them as alternative fumigants in seed stores.     

Efficacy of crude neem seed kernel oil (NSKO) in controlling the tree locust (Anacridium melanorhodon melanorhodon Walker), a serious pest of the gum arabic producing tree (Acacia senegal L. Willd.) in the Sudan

The present study was carried out at Al Semih (western Sudan) (13° 14′ N: 30° 27′ E: Alt. 550 m). The area is part of the gum arabic belt which is heavily populated with farmers who practice shifting cultivation and/or agroforestry where Acacia senegal L. Willd (the main gum arabic producing tree) is dominant. The tree locust (Anacridium melanorhodon melanorhodon Walker (Orthoptera, Acrididae) is a serious pest of A. senegal. Farmers practice agroforestry within the gum belt where they cultivate their crops e.g. millet, groundnut, sesame and sorghum between A. senegal trees. Gum Arabic is an important source of revenue for farmers. Chemical pesticides have been used to control the tree locust. Although the gum belt (where A. senegal grows) is heavily populated, pesticides are used to control the tree locust in the area. The present study was undertaken with the objective of using alternative environmentally safe pesticides. Several doses of neem seed kernel crude oil (NSKO) were tested against the 4^th, 5^th, 6^th instars and the mature tree locust (A. melanorhodon melanorhodon) in a completely randomized block design. Experiments were carried out in cages placed on part of the gum arabic belt. NSKO at high doses (5 and 10% v/v) significantly reduced feeding, moulting and ovipositing of the tree locust and significantly increased the mortality of all developmental stages tested. However, low doses of NSKO had no significant effect on the tree locust.     

Mutations in aARS genes revealed by targeted next-generation sequencing in patients with mitochondrial diseases

Molecular Biology Reports - Mitochondrial diseases are a clinically heterogeneous group of multisystemic disorders that arise as a result of various mitochondrial dysfunctions. Autosomal recessive...     

Synthesis and characterization of a series of N3O-ligated mononuclear Mn(II) halide complexes

Treatment of a N₃O-donor chelate ligand (mpppa = N-methyl-N-((6-pivaloylamido-2-pyridyl)methyl)-N-(2-pyridylethyl)amine; bpppa = N-benzyl-N-((6-pivaloylamido-2-pyridyl)methyl)-N-(2-pyridylmethyl)amine) with equimolar amounts of Mn(ClO₄)₂ · 6H₂O and Me₄NX (X = Cl, Br, I) in methanol resulted in the production of a series of mononuclear Mn(II) halide complexes of the formula [(L)Mn-X(CH₃OH)]ClO₄ (L = mpppa or bpppa). X-ray crystallographic studies of [(mpppa)Mn-Cl(CH₃OH)]ClO₄ · CH₃OH (2 · CH₃OH), [(mpppa)Mn-Br(CH₃OH)]ClO₄ · CH₃OH (4 · CH₃OH), [(mpppa)Mn-I(CH₃OH)]ClO₄ · CH₃OH (6 · CH₃OH), and [(bpppa)Mn-I(CH₃OH)]ClO₄ · O₂(CH₂CH₃)₂ (7 · O(CH₂CH₃)₂) revealed for each a mononuclear Mn(II) center having tetradentate coordination of the chelate ligand, one coordinated halide anion, and one molecule of coordinated methanol. An increase in the Mn-X distance through the halide series (Cl, Br, I) correlates linearly with the increase in the radius of the anion. The magnetic moment of each halide complex, measured via Evans method in methanol, is consistent with the presence of a high-spin distorted octahedral Mn(II) center. The EPR features of the halide complexes in methanol do not change as a function of the nature of the halide coordinated to the Mn(II) center.     

The c.242G>A mutation in LRTOMT gene is responsible for a high prevalence of deafness in the Moroccan population

Congenital hearing impairment (HI) affects one in 1,000 newborns and has a genetic cause in 50?% of the cases. Autosomal recessive non-syndromic hearing impairment is responsible for 70–80?% of all hereditary cases of HI. Recently, it has been demonstrated that, mutations of LRTOMT are associated with profound nonsyndromic hearing impairment at the DFNB63 locus. The objective of this study is to evaluate the carrier frequency of c.242G>A mutation in LRTOMT gene and define the contribution of this gene in the etiology of deafness in Moroccan population. We screened 105 unrelated Moroccan families with non-syndromic HI and 120 control individuals for mutation in the exon 8 of the LRTOMT gene, by sequencing and PCR-RFLP. The Homozygous c.242G>A mutation was found in 8.75?% of the families tested and in 4.16?% of control in the heterozygous state. Our results show that after the GJB2 gene mutation in LRTOMT gene is the second cause of congenital hearing impairment in Moroccan patients. This finding should facilitate diagnosis of congenital deafness of the affected subjects in Morocco.     

Using community and population approaches to understand how contemporary and historical factors have shaped species distribution in river ecosystems

Aim To examine how the employment of both community‐ and population‐level approaches can provide a wider view of the importance of contemporary and historical factors on current species distribution. We posit that community ecology should provide more information about contemporary factors, whereas population genetics should provide better information about historical factors. Location Rivers of the western Mediterranean Basin, including four subregions differing in geological history: the Iberian Plate, Transitional, Betic and Rif. Methods For a community‐level approach, Trichoptera richness and community composition were compared between subregions using species accumulation curves and a correspondence analysis. For a population‐level approach, the mtDNA cytochrome C oxidase subunit I (COI) gene of specimens of the Trichoptera midstream‐lowland species Chimarra marginata (L.) was sequenced and analysed using phylogeographical methods. Results The community approach revealed that historical events had more influence on headwater communities than contemporary ecological factors, whereas historical events had negligible influence on midstream‐lowland communities. In midstream‐lowland sites, however, the population approach showed that the genetic structure of C. marginata differed significantly between subregions and revealed patterns of historical gene migration. In terms of species richness, the Rif subregion had the lowest value per basin due to local climatic features and isolation. Main conclusions Both community‐ and population‐level approaches yielded information about the effects of historical factors on species distribution. However, the importance of historical events on current Trichoptera communities depends on the river zonation. Unlike headwater sites, midstream‐lowland sites showed signs of historical events at the population level but not at the community level at the scale used, indicating that both approaches should be employed together in biogeographical studies. Lack of detection of historical events at the community level does not necessarily mean that they are negligible. Most likely, the organizational level used is not appropriate. We also stress the importance of implementing conservation measures for rivers in the western Mediterranean, especially under future scenarios of climate change and human disturbances in the Mediterranean Basin.