ISSR fingerprinting of Coffea arabica throughout Ethiopia reveals high variability in wild populations and distinguishes them from landraces

Forests of SW Ethiopia constitute the native habitat of Coffea arabica and also the place where domestication of Arabica coffee started. Selection from wild populations has led to numerous landraces (farmer’s varieties) and cultivars. Inter-simple sequence repeats (ISSRs) were generated from a representative set of forest coffee populations and landraces across Ethiopia. For the broad diversity assessment, nine di- and tri-nucleotide ISSR primers were applied, as chosen from a total of 102 primers tested initially. Tetranucleotide ISSR primers differed in amplifying fingerprints that could hardly be analysed due to excessive variation. Tree building analysis (NJ, UPGMA) of 84 polymorphic loci amplified for 125 C. arabica individuals provided evidence for several groups of related genotypes occurring in certain geographical areas of Ethiopia and underscored the existence of wild coffee distinct from landraces. Landraces seem to have originated in different geographical areas of Ethiopia in a stepwise domestication process. While the overall geographical signal in the dataset was weak, analysis in a Bayesian framework using the admixture model with geographical priors in STRUCTURE recovered some genetic clustering. Based on Shannon’s diversity index, populations from Yayu (0.47) and Bonga (0.46) showed highest diversity, followed by individuals from Berhane Kontir (0.41). A likely scenario for the differentiation of C. arabica after an allopolyploidization event is that the hierarchical-geographical patterning of wild Coffea genotypes expected from stepwise range extension was obscured by recent or ancient gene flow. The diversity and geographical distribution of autochthonous C. arabica genotypes indicates the need for a multi-site in situ conservation approach.     

CAL1 is the Drosophila CENP-A assembly factor

Centromeres are specified epigenetically by the incorporation of the histone H3 variant CENP-A. In humans, amphibians, and fungi, CENP-A is deposited at centromeres by the HJURP/Scm3 family of assembly factors, but homologues of these chaperones are absent from a number of major eukaryotic lineages such as insects, fish, nematodes, and plants. In Drosophila, centromeric deposition of CENP-A requires the fly-specific protein CAL1. Here, we show that targeting CAL1 to noncentromeric DNA in Drosophila cells is sufficient to heritably recruit CENP-A, kinetochore proteins, and microtubule attachments. CAL1 selectively interacts with CENP-A and is sufficient to assemble CENP-A nucleosomes that display properties consistent with left-handed octamers. The CENP-A assembly activity of CAL1 resides within an N-terminal domain, whereas the C terminus mediates centromere recognition through an interaction with CENP-C. Collectively, this work identifies the “missing” CENP-A chaperone in flies, revealing fundamental conservation between insect and vertebrate centromere-specification mechanisms.     

Species composition,abundance and habitat association of rodents and insectivores from Chilalo–Galama Mountain range,Arsi, Ethiopia

A study on the ecology of rodents and insectivores on Mount Chilalo and the Galama Mountain range was conducted during August, 2007–April, 2008. Study grids were marked in nine randomly selected habitats. Sherman live traps and snap traps were used to capture the small mammals. A total of 2525 (including recaptures) individual rodents and insectivores were captured in 5488 (Sherman) and 1590 (snap) trap‐nights. Among the live‐trapped animals, 2037 (96.2%) and 80 (3.8%) were rodents and insectivores, respectively, that comprised 17 species. These were as follows: Arvicanthis abyssinicus, Arvicanthis blicki, Arvicanthis dembeensis, Lophuromys flavopunctatus, Lophuromys melanonyx, Mastomys natalensis, Mus mahomet, Otomys typus, Rattus rattus, Stenocephalemys albipes, Stenocephalemys albocaudata, Stenocephalemys griseicauda, Crocidura glassi, Crocidura flavescens and Crocidura thalia. Tachyoryctes splendens and Hystrix cristata were sighted but not captured. Endemics to Ethiopia constituted 60% of the trapped small mammals. Lophuromys flavopunctatus was the most abundant and widely distributed rodent species, while M. mahomet was the least. The altitudes between 3301 and 3400 m asl had the highest species composition and abundance although it is second in number of catches per trap‐nights. The majority of rodents and insectivores were associated with montane forest habitat, whereas the least were in the shrubland. Anthropogenic effects were witnessed affecting the distribution of small mammals.     

Capillary isoelectric focusing coupled with dynamic imaging detection: A one-dimensional separation for two-dimensional protein characterization

Here, we devised a novel approach for two-dimensional (2D) protein characterization using a single one-dimensional separation followed by a second characterization in the same instrument. The approach combines capillary isoelectric focusing (CIEF), which separates proteins according to pI, with dynamic imaging detection, which permits monitoring of protein diffusion in real time and thereby allows estimation of molecular weight from diffusion coefficient. Compared with classical 2D gel electrophoresis, the approach provided several significant advantages including speed and ease in operation and automation, while yielding comparable accuracy. The approach was applicable for protein samples of low to moderate complexity.     

Genome-wide DNA methylation profiling of chronic lymphocytic leukemia allows identification of epigenetically repressed molecular pathways with clinical impact

We performed a genome-wide analysis of aberrant DNA methylation in chronic lymphocytic leukemia (CLL) using methylated CpG island amplification (MCA) coupled with a promoter microarray. We identified 280 potential targets of aberrant DNA methylation in CLL. These genes were located more frequently in chromosomes 19 (16%, p = 0.001), 16 (11%, p = 0.001), 17 (10%, p = 0.02) and 11 (9%, p = 0.02) and could be grouped in several functional networks. Methylation status was confirmed for 22 of these genes (SOX11, DLX1, FAM62C, SOX14, RSPO1, ADCY5, HAND2, SPOCK, MLL, ING1, PRIMA1, BCL11B, LTBP2, BNC1, NR2F2, SALL1, GALGT2, LHX1, DLX4, KLK10, TFAP2 and APP) in 78 CLL patients by pyrosequencing. As a proof of principle, we analyzed the expression of 2 genes, PRIMA1 and APP, in primary cells and of GALGT2, TFAP2C and PRIMA1 in leukemia cells. There was an inverse association between methylation and gene expression. This could be reversed by treatment with 5-aza-2′-deoxycytidine in cell lines. Treatment in a clinical trial with 5-azacitidine resulted in decreased methylation of LINE, DLX4 and SALL1 in the peripheral blood B-cells of patients with CLL. IgV_H mutational status or ZAP-70 expression were not associated with specific methylation profiles. By multivariate analysis, methylation of LINE and APP was associated with shorter overall survival (p = 0.045 and 0.0035, respectively). This study demonstrates that aberrant DNA methylation is common and has potential prognostic and therapeutic value in CLL.Key words: chronic lymphocytic leukemia, DNA methylation, MCA/promoter microarray, epigenetics     

Phylogenetics and taxonomic delimitation of the genus Guizotia (Asteraceae) based on sequences derived from various chloroplast DNA regions

Parsimony-based phylogenetic analyses of the genus Guizotia were undertaken based on DNA sequence data from the following chloroplast DNA (cpDNA) regions: trnT-trnL, trnL-trnF, trnY-rpoB, trnC-petN, psbM-trnD and rps16-trnQ intergenic spacers, trnL, rps16 and matK-5′trnK introns and matK gene. Out of the 26 primers used in this study, 14 were newly designed. The study was conducted to determine (1) the closest relative of Guizotia abyssinica, (2) the taxonomic status of some Guizotia taxa and (3) the subtribal placement of Guizotia in the tribe Heliantheae. The analyses of the sequence data showed that G. abyssinica, G. scabra ssp. scabra, G. scabra ssp. schimperi and G. villosa are phylogenetically closely related. However, G. scabra ssp. schimperi appeared as the most closely related taxon to G. abyssinica. Based on this phylogenetic analysis, we suggest that the two subspecies of G. scabra are better treated as separate species. The analysis also clearly demonstrated that “Chelelu” and “Ketcha” are distinct Guizotia species. The trnT-trnL and trnL-trnF intergenic spacer-based phylogenetic analysis of various subtribes of the tribe Heliantheae strongly supports the placement of the genus Guizotia within the subtribe Milleriinae.     

Vision Loss and Psychological Distress among Ethiopians Adults: A Comparative Cross-Sectional Study

Background

Vision loss causes major changes in lifestyle and habits that may result in psychological distress and further reduction in the quality of life. Little is known about the magnitude of psychological distress in patients with vision loss and its variation with the normal. The aim of this study is, therefore, to investigate the psychological effects of vision loss and its determinants among Ethiopians.

Methods

A comparative cross-sectional study was conducted on adults attending the Eye clinic of Jimma University Hospital. One hundred fifteen consecutive adults with visual loss at least in one eye and 115 age-and sex-matched controls with normal vision were studied. The psychological distress was measured using standardized Self-Reporting Questionnaire (SRQ-20). Chi-square test and logistic regression were carried out and associations were considered significant at P<0.05.

Results

The overall prevalence of psychological distress was 33.4%. While psychological distress was found in 49.8% of patients who had loss of vision at least in one eye, only 18.3% of the controls had it. In the adjusted analysis, patients with vision loss had 4.6 times higher risk of suffering from psychological distress compared to patients with normal vision (AOR 4.56; 95% CI 2.16-9.62). Moreover, patients with vision loss in both eyes (AOR 4.00; 95% CI 1.453-11.015) and with worse visual acuity in the better eye (AOR 3.66; 95% CI 1.27-10.54) were significantly more likely to have psychological distress than those patients with vision loss in one eye only and good visual acuity in the better eye respectively. The cause of visual loss, pattern of visual loss, duration of visual loss and sociodemographic variables did not influence the likelihood of having psychological distress.

Conclusion

Prevalence of psychological distress was significantly higher in patients with visual loss compared to patients with normal vision. There is a need for integration of psychosocial care into the current medical and surgical treatment of patients with vision loss.     

<Emphasis Type="Italic">Camelus dromedarius</Emphasis> brucellosis and its public health associated risks in the Afar National Regional State in northeastern Ethiopia

Background

A cross-sectional study was carried out in four districts of the Afar region in Ethiopia to determine the prevalence of brucellosis in camels, and to identify risky practices that would facilitate the transmission of zoonoses to humans. This study involved testing 461 camels and interviewing 120 livestock owners. The modified Rose Bengal plate test (mRBPT) and complement fixation test (CFT) were used as screening and confirmatory tests, respectively. SPSS 16 was used to analyze the overall prevalence and potential risk factors for seropositivity, using a multivariable logistic regression analysis.

Results

In the camel herds tested, 5.4% had antibodies against Brucella species, and the district level seroprevalence ranged from 11.7% to 15.5% in camels. The logistic regression model for camels in a herd size?>?20 animals (OR?=?2.8; 95% CI: 1.16-6.62) and greater than four years of age (OR?=?4.9; 95% CI: 1.45-16.82) showed a higher risk of infection when compared to small herds and those?≤?4 years old. The questionnaire survey revealed that most respondents did not know about the transmission of zoonotic diseases, and that their practices could potentially facilitate the transmission of zoonotic pathogens.

Conclusions

The results of this study revealed that camel brucellosis is prevalent in the study areas. Therefore, there is a need for implementing control measures and increasing public awareness in the prevention methods of brucellosis.

     

Phylogenetic diversity and dietary association of rumen Treponema revealed using group-specific 16S rRNA gene-based analysis

Treponema spp. are a commonly detected bacterial group in the rumen that are involved in the degradation of soluble fibers. In this study, a ruminal Treponema group-specific PCR primer targeting the 16S rRNA gene was designed and used to assess the phylogenetic diversity and diet association of this group in sheep rumen. Total DNA was extracted from rumen digesta of three sheep fed a diet based on alfalfa/orchardgrass hay or concentrate. The real-time PCR quantification indicated that the relative abundance of the Treponema group in the total rumen bacteria was as high as 1.05%, while the known species Treponema bryantii accounted for only 0.02%. Fingerprints of the Treponema community determined by 16S rDNA-targeted denaturing gradient gel electrophoresis (DGGE) analysis tended to differ among the diets. Principal component analysis of the DGGE profiles distinguished those Treponema associated with either the hay or the concentrate diets. Analysis of a Treponema 16S rRNA gene clone library showed phylogenetically distinct operational taxonomic units for a specific dietary condition, and significant (P=0.001) differences in community composition were observed among clone libraries constructed from each dietary regimen. The majority of clones (75.4%) had <97% sequence similarity with known Treponema. These results suggest the predominance of uncultured Treponema that appear to have distinct members related to the digestion of either hay or concentrate diet.