New sources of dwarfing genes in pearl millet (Pennisetum americanum)

Summary Thirteen naturally occurring dwarf lines of pearl millet [Pennisetum americanum (L.) Leeke], identified from the world collection, varied for several morphological and agronomic characters. Extreme dwarfs were characterized by a tufted growth habit which could be distinguished from the time of germination, while the other dwarf lines could be distinguished only after anthesis. The F₁ hybrids between the tall and dwarf genotypes were tall, indicating that dwarfness is a recessive trait. In 10 out of the 13 crosses, the F₂ segregation ratio was three tall to one dwarf (31) suggesting that the dwarfness is controlled by a single recessive gene, while the height differences in 3 of the dwarfs (IP 8056, IP 8210 and IP 8214) were controlled by more than one gene as they showed continuous variation for plant height in F₂. When the remaining 10 single gene dwarfs were crossed to either d ₁ (Tift 238) or d ₂ (Tift 23 DB) dwarfs, only 2 crosses produced tall F₂ hybrids and they segregated for height in F₂ indicating that these 2 dwarfs are non-allelic to d ₁ and d ₂. Reciprocal crosses of these 2 dwarfs produced tall F₁ hybrids and showed a dihybrid segregation of 934 in F₂ indicating that the dwarfing genes of these 2 parents are non-allelic to each other. These non-allelic dwarfs were assigned the gene symbols d ₃ (IP 10401), and d ₄ (IP 10402).Submitted as J.A. No. 429 by the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT)     

Simple Chromatographic Method for Simultaneous Analyses of Phosphatidylcholine, Lysophosphatidylcholine, and Free Fatty Acids

This study describes a simple chromatographic method for the simultaneous analyses of phosphatidylcholine (PC) and its hydrolytic degradation products: lysophosphatidylcholine (LPC) and free fatty acids (FFA). Quantitative determination of PC, LPC, and FFA is essential in order to assure safety and to accurately assess the shelf life of phospholipid-containing products. A single-run normal-phase high-performance liquid chromatography (HPLC) with evaporative light scattering detector has been developed. The method utilizes an Allsphere silica analytical column and a gradient elution with mobile phases consisting of chloroform: chloroform–methanol (70:30%, v/v) and chloroform–methanol–water–ammonia (45:45:9.5:0.5%, v/v/v/v). The method adequately resolves PC, LPC, and FFA within a run time of 25 min. The quantitative analysis of PC and LPC has been achieved with external standard method. The free fatty acids were analyzed as a group using linoleic acid as representative standard. Linear calibration curves were obtained for PC (1.64–16.3 μg, r² = 0.9991) and LPC (0.6–5.0 μg, r² = 0.9966), while a logarithmic calibration curve was obtained for linoleic acid (1.1–5.8 μg, r² = 0.9967). The detection and quantification limits of LPC and FFA were 0.04 and 0.1 μg, respectively. As a means of validating the applicability of the assay to pharmaceutical products, PC liposome was subjected to alkaline hydrolytic degradation. Quantitative HPLC analysis showed that 97% of the total mass balance for PC could be accounted for in liposome formulation. The overall results show that the HPLC method could be a useful tool for chromatographic analysis, stability studies, and formulation characterization of phospholipid-based pharmaceuticals.KEY WORDS: evaporative light scattering detection, free fatty acid, lysophosphatidylcholine, phosphatidylcholine     

Human TRMU encoding the mitochondrial 5-methylaminomethyl-2-thiouridylate-methyltransferase is a putative nuclear modifier gene for the phenotypic expression of the deafness-associated 12S rRNA mutations

Nuclear modifier genes have been proposed to modulate the phenotypic manifestation of human mitochondrial 12S rRNA A1491G mutation associated with deafness in many families world-wide. Here we identified and characterized the putative nuclear modifier gene TRMU encoding a highly conserved mitochondrial protein related to tRNA modification. A 1937bp TRMU cDNA has been isolated and the genomic organization of TRMU has been elucidated. The human TRMU gene containing 11 exons encodes a 421 residue protein with a strong homology to the TRMU-like proteins of bacteria and other homologs. TRMU is ubiquitously expressed in various tissues, but abundantly in tissues with high metabolic rates including heart, liver, kidney, and brain. Immunofluorescence analysis of human 143B cells expressing TRMU-GFP fusion protein demonstrated that the human Trmu localizes and functions in mitochondrion. Furthermore, we show that in families with the deafness-associated 12S rRNA A1491G mutation there is highly suggestive linkage and linkage disequilibrium between microsatellite markers adjacent to TRMU and the presence of deafness. These observations suggest that human TRMU may modulate the phenotypic manifestation of the deafness-associated mitochondrial 12S rRNA mutations.     

Stiffness of desiccating insect wings

The stiffness of insect wings is typically determined through experimental measurements. Such experiments are performed on wings removed from insects. However, the wings are subject to desiccation which typically leads to an increase in their stiffness. Although this effect of desiccation is well known, a comprehensive study of the rate of change in stiffness of desiccating insect wings would be a significant aid in planning experiments as well as interpreting data from such experiments. This communication presents a comprehensive experimental analysis of the change in mass and stiffness of gradually desiccating forewings of Painted Lady butterflies (Vanessa?cardui). Mass and stiffness of the forewings of five butterflies were simultaneously measured every 10 min over a 24 h period. The averaged results show that wing mass declined exponentially by 21.1% over this time period with a time constant of 9.8 h, while wing stiffness increased linearly by 46.2% at a rate of 23.4 μN mm(-1) h(-1). For the forewings of a single butterfly, the experiment was performed over a period of 1 week, and the results show that wing mass declined exponentially by 52.2% with a time constant of 30.2 h until it reached a steady-state level of 2.00 mg, while wing stiffness increased exponentially by 90.7% until it reached a steady-state level of 1.70 mN mm(-1).     

Genetic and non-genetic parameter estimates for growth traits and Kleiber ratios in Dorper × indigenous sheep

Genetic improvement programme will only be successful when accompanied by a good understanding of the influence of different environmental factors, knowledge of the genetic parameters and the genetic relationships between the traits of interest. This study aimed to evaluate the influence of non-genetic factors on growth traits and Kleiber ratios and to estimate genetic parameters for early growth traits in Dorper × indigenous crossbred sheep. The effects of fixed factors were analysed by the general linear model procedure of statistical analysis system, while the genetic parameters were estimated using a WOMBAT computer program fitted animal model. The overall least-square mean for birth weight (BRW), weaning weight (3MW), six-month weight, nine-month weight, and yearling weight were 3.03 ± 0.02, 14.5 ± 0.18, 20.4 ± 0.26, 24.8 ± 0.31, and 28.3 ± 0.40 kg, respectively. The overall least-square mean for Kleiber ratio from birth to weaning (KR1), weaning to six months, six to nine months and nine months to yearling age were 16.8 ± 0.10, 6.41 ± 0.17, 4.55 ± 0.21 and 3.38 ± 0.20 g/kg of metabolic weight, respectively. The inclusion of maternal genetic effect had a significant influence on BRW, and it explains 20% of the phenotypic variation. The total heritability estimates for BRW, 3MW, birth to weaning average daily weight gain and KR1 were 0.10, 0.14, 0.16 and 0.12, respectively. The phenotypic correlation varied from ?0.11 ± 0.05 to 0.98 ± 0.02, whereas the direct genetic correlation ranged from ?0.32 ± 0.40 to 0.98 ± 0.17. The mean inbreeding coefficient was 0.105% with an annual rate of 0.02%. The heritability estimates for growth traits and Kleiber ratio suggest that slow genetic progress would be expected from the selection. However, the integration of selection with crossbreeding programme can enhance genetic gain. Therefore, selection should be conducted based on breeding values estimated from multiple information sources to increase the selection response.     

Mitochondrial myopathy and sideroblastic anemia (MLASA): missense mutation in the pseudouridine synthase 1 (PUS1) gene is associated with the loss of tRNA pseudouridylation

A missense mutation in the PUS1 gene affecting a highly conserved amino acid has been associated with mitochondrial myopathy and sideroblastic anemia (MLASA), a rare autosomal recessive oxidative phosphorylation disorder. The PUS1 gene encodes the enzyme pseudouridine synthase 1 (Pus1p) that is known to pseudouridylate tRNAs in other species. Total RNA was isolated from lymphoblastoid cell lines established from patients, parents, unaffected siblings, and unrelated controls, and the tRNAs were assayed for the presence of pseudouridine (Psi) at the expected positions. Mitochondrial and cytoplasmic tRNAs from MLASA patients are lacking modification at sites normally modified by Pus1p, whereas tRNAs from controls, unaffected siblings, or parents all have Psi at these positions. In addition, there was no Pus1p activity in an extract made from a cell line derived from a patient with MLASA. Immunohistochemical staining of Pus1p in cell lines showed nuclear, cytoplasmic, and mitochondrial distribution of the protein, and there is no difference in staining between patients and unaffected family members. MLASA is thus associated with absent or greatly reduced tRNA pseudouridylation at specific sites, implicating this pathway in its molecular pathogenesis.     

Impact of the Use of a Rapid Diagnostic Test for Visceral Leishmaniasis on Clinical Practice in Ethiopia: A Retrospective Study

Background

Diagnostic guidelines for Visceral Leishmaniasis (VL) in the East African region are complex. Patients meeting the VL clinical case definition should be tested by rK39 rapid diagnostic test (RDT) followed by the Direct Agglutination Test (DAT) or tissue aspiration if RDT-negative. Otherwise, RDT-positive patients should be started on VL treatment. We evaluated how this guideline is adhered to by assessing the routine clinical practice in a university hospital in North-West Ethiopia.

Methods

Retrospective record analysis was done for all patients who had an rK39-RDT done at University of Gondar (UoG) Hospital between June 2012 and June 2013. We described the diagnostic work-up performed and the proportion initiated on VL treatment by test result.

Results/Findings

From a total of 928 patients tested, 308 (33.2%) were rK39 RDT-positive. Spleen or bone marrow aspiration was done for 237 (77.2%) RDT-positive patients. Of these, 165 were confirmed parasitologically, yielding a positive predictive value of 69.6%. Only 126 (20.3%) of the 620 patients with a negative rK39 test underwent further testing by tissue aspiration, of which 22 (17.5%) were also parasitology positive. HIV test results were available for 570 (61.4%) patients and 36 (6.3%) were HIV-infected. Of the 187 parasitologically confirmed patients, 182 (97.3%) were started on VL treatment.

Conclusions / Discussion

A negative rK39 test was often not followed by further testing and a positive rK39 test result was followed by tissue aspiration in three out of four cases. Further research is required to understand why the diagnostic work-up did not comply with the guidelines, including evaluating adherence to the VL clinical case definition and quality of rK39-RDT testing.     

Environment differentially affects the functional and phylogenetic structures of plant communities in a dry evergreen Afromontane tropical forest

Testing how local environmental conditions influence plant community assembly is important to understand the underlying mechanisms that promote and/or maintain biodiversity. Functional traits are used to find the broad spectrum of resource use strategies that plants use to respond to environmental variation. The patterns and drivers of plant community assembly through the lens of traits and phylogeny; however, remain to be studied in a uniquely biodiversity rich but poorly known fragmented dry Afromontane forest of Ethiopia. Here, we combined trait and community phylogenetic data from thirty sampling plots of 20 × 20 m size to determine the functional and phylogenetic structures and their drivers in a fragmented, human-dominated dry evergreen Afromontane forest. We found phylogenetic and functional clustering of plants in which the effect of environment was found to be trait specific. A weak phylogenetic signal for traits was detected suggesting that species resource use strategies may not be inferred using species phylogenetic distance. Additionally, we found functional traits to be weak in predicting species abundance distribution. Overall, while this study shows a non-random community assembly pattern, it also highlights the importance of deterministic processes being trait specific.