A comparison of genetic stability in tea [Camellia sinensis (L.) Kuntze] plantlets derived from callus with plantlets from long-term in vitro propagation

Plant Cell, Tissue and Organ Culture (PCTOC) - The development of tissue-specific or inducible promoters is important for plant genetic engineering. In this study, we isolated two novel promoters...     

The genetic structure of cattle populations (Bos taurus) in northern Eurasia and the neighbouring Near Eastern regions: implications for breeding strategies and conservation

We investigated the genetic structure and variation of 21 populations of cattle (Bos taurus) in northern Eurasia and the neighbouring Near Eastern regions of the Balkan, the Caucasus and Ukraine employing 30 microsatellite markers. By analyses of population relationships, as well as by a Bayesian-based clustering approach, we identified a genetic distinctness between populations of modern commercial origin and those of native origin. Our data suggested that northern European Russia represents the most heavily colonized area by modern commercial cattle. Further genetic mixture analyses based on individual assignment tests found that native Red Steppe cattle were also employed in the historical breeding practices in Eastern Europe, most probably for incorporating their strong and extensive adaptability. In analysis of molecular variance, within-population differences accounted for approximately 90% of the genetic variation. Despite some correspondence between geographical proximity and genetic similarity, genetic differentiation was observed to be significantly associated with the difference in breeding purpose among the European populations (percentage of variance among groups and significance: 2.99%, P = 0.02). Our findings give unique genetic insight into the historical patterns of cattle breeding practices in the former Soviet Union. The results identify the neighbouring Near Eastern regions such as the Balkan, the Caucasus and Ukraine, and the isolated Far Eastern Siberia as areas of 'genetic endemism', where cattle populations should be given conservation priority. The results will also be of importance for cost-effective management of their future utilization.     

Development of enzyme immunoassay of amphetamines in human biological fluids

Enzyme-linked immunosorbent assay (ELISA) for detection of amphetamines in urine was developed. The conditions for ELISA using an antigen obtained from the derivative of amphetamine and ovalbumin were optimized. It was shown that the specificity of the proposed method when used in clinical conditions corresponds to that of gas chromatography with mass spectrometric detection.     

Genetic differentiation between the Old and New types of Serbian Tsigai sheep

Two Tsigai sheep populations exist in Serbia: the Old type, called Čokan, and the New type. It is assumed that the New type results from upgrading Tsigai sheep with exotic genetic material. We investigated genetic diversity and differentiation of these types by analysing 23 autosomal microsatellites. Tests for Hardy-Weinberg proportions, linkage equilibrium between genotypes across loci and the calculation of inbreeding coefficients were performed and the deficiency in the number of alleles within the Tsigai types was examined using a Wilcoxon sign-rank test. The New type displayed a higher level of genetic variability than the Čokan in terms of allele numbers, but the New Tsigai showed a pattern of heterozygosity deficiency. The positive f value for the Čokan suggests the occurrence of inbreeding in this type. The proportion of linkage disequilibrium was below that expected by chance. Exclusion of two loci in Hardy-Weinberg disequilibrium did not alter our conclusions based on the entire data set i.e. the two Tsigai types are clearly differentiated and the New Tsigai type has been influenced by crossbreeding. Therefore, the Čokan Tsigai should be considered as a distinct endangered breed in the FAO classification.     

Estimation of breed contributions to present and future genetic diversity of 44 North Eurasian cattle breeds using core set diversity measures

Extinction of breeds threatens genetic diversity of livestock species. The need to conserve genetic diversity is widely accepted but involves in general two questions: (i) is the expected loss of diversity in a set of breeds within a defined future time horizon large enough to establish a conservation plan, and if so (ii) which breeds should be prioritised for such a conservation plan? The present study uses a marker assisted methodology to address these questions. The methodology combines core set diversity measures with a stochastic method for the estimation of expected future diversity and breed marginal diversities. The latter is defined as the change in the total diversity of all breeds caused by a one unit decrease in extinction probability of a particular breed. The stochastic method was validated by means of simulations. A large field data set consisting of 44 North Eurasian cattle breeds was analysed using simplified determined extinction probabilities. The results show that the expected loss of diversity in this set within the next 20 to 50 years is between 1 and 3% of the actual diversity, provided that the extinction probabilities which were used are approximately valid. If this loss is to be reduced, it is sufficient to include those three to five breeds with the highest marginal diversity in a conservation scheme.     

A phylogenetic study of Dermestidae (Coleoptera) based on larval morphology

Abstract.  The results of a phylogenetic analysis of Dermestidae (Coleoptera) based on the morphology of immature stages are presented and implications for the classification of the family are discussed. The monophyly of (( Orphilus ) + (Dermestidae) + (Endecatomidae (Bostrichidae + Anobiidae))) is strongly supported. Dermestidae, excluding Orphilus Erichson, forms a monophyletic group. The phylogenetic position of the Orphilinae remains enigmatic. The hypothesized relationships of the remaining dermestid subfamilies are (Thorictinae (Dermestinae (Attageninae (Megatominae + Trinodinae)))). Phylogenetically informative morphological characters of larvae and pupae are illustrated and described. Character state transformations implied by the phylogenetic hypothesis are discussed. New morphological observations are provided, including the first records of the presence of gastric caeca in the larva of Dermestes L., and the first report of the number of Malpighian tubules and the number of abdominal ganglia in Dermestidae, Bostrichidae, Anobiidae, Nosodendridae and Derodontidae. The form of the mandibular base is described for the first time for Thylodriini Beal and Trinodini Beal. The fossil record and distribution of extant Dermestidae are discussed in the light of the phylogenetic hypothesis. The origin of the family is hypothesized to be in the Late Cretaceous.     

Plasma and saliva miR-21 expression in colorectal cancer patients

MicroRNA-21 (miR-21) expression was quantified by real-time qRT-PCR in peripheral blood and saliva samples obtained from patients diagnosed with colorectal cancer (CRC) of varying degrees of malignancy and healthy volunteers. All patients had adenocarcinoma located in the distal colon at different stages. Significant differences were detected between the control group and the total experimental group of CRC patients (plasma, P?=?0.0001; saliva, P?=?5e-12). MiR-21 expression was also significantly different in certain subgroups of patients with CRC disease stages II–IV as compared to the control group. No correlation of miR-21 expression was found with regard to gender and age of patents. Also, there were no significant individual correlations and linear regression of miR-21 expression in the plasma and saliva. The estimated diagnostic sensitivity and specificity of miR-21 expression were respectively 65 and 85% in the plasma, and 97 and 91% in the saliva. Our data suggest that miR-21 in both the saliva and plasma could be a proper biomarker for CRC screening, although the saliva miR-21 expression test looks preferable due to its higher sensitivity, specificity, and technical simplicity.     

Linkage disequilibrium analysis for microsatellite loci in six cattle breeds

Autosomal microsatellites are valuable tools for investigating genetic diversity and population structure and making conservation decisions to preserve valuable breeds of domestic animals. We carried out a linkage disequilibrium analysis using 29 microsatellite markers in six cattle populations: Suksun, Istoben, Yaroslavl, Kholmogory, Grey Ukrainian and Pechora type of Khologory cattle. We discovered a significant linkage disequilibrium between microsatellites INRA037 and CSRM60 in Grey Ukrainian cattle.     

Immunoaffinity chromatography of human thyroid peroxidase: The stability of the three-dimensional structure and immunoreactivity of antigen and antibodies under various elution conditions

Actions of various chemical agents modeling immunoaffinity chromatography elution conditions caused structural changes of the components of human thyroid peroxidase (TPO) complexes with monoclonal antibodies (MABs) F8 and A1 whose antigenic determinants have a conformational nature and are located in the immunodominant region and a peripheral region of TPO, respectively. These changes became apparent in the circular dichroism and fluorescence spectra of TPO and both MABs as well as in the immunoassay. The effectiveness of the chemical reagents with respect to TPO desorption from an immobilized MAB decreased in the following order: 0.2 M ammonia (pH 11.5) > 0.1 M lithium 3,5-diiodosalycilate > 0.1 M glycine-HCl (pH 2.5) > 1 M NaI > 30% propylene glycol + 1 M NaCl > 30% propylene glycol > 1 M NaCl. At pH 11.5, the three-dimensional structure and immunoreactivity of TPO retained completely and only minor alterations of MAB analogical parameters took place, thus providing a high yield of the functional active human TPO and favoring repeated use of the immobilized MABs in immunoaffinity chromatography. The results may be used as a strategy for the optimization of various protein antigens immunoaffinity chromatography.