Witold Stanisław Proskura

Synthesis and structural X-ray analysis of 1,1’-(naphthalene-1,8-diyl)-3,3’-dibenzoyl-bisthiourea and its use as anion-binding receptor

* Correspondence: witold.proskura@zut.edu.pl Sterol regulatory element-binding protein 1 (SREBP1) is the main transcription factor linked to lipogenesis regulation. SREBP1 regulates the expression of the FASN and ACACA genes coding for the 2 major lipogenic enzymes involved in de novo fatty acid synthesis, fatty acid synthase and acetyl-CoA carboxylase, respectively (1). The SREBP1 pathway plays a key role in the regulation of milk fat synthesis, and thus the variation within the SREBP1 gene may influence fat content and especially fatty acid composition in bovine milk and meat (2). Hoashi et al. (3) determined the complete sequence of this gene. They discovered an insertion/deletion (ins/del) of 84 bp in intron 5 of the SREBP1 gene. The short (S)and long (L)-type alleles were identified. Recently, several studies have indicated an association between this ins/ del polymorphism and fatty acid composition in the milk and meat of cattle (2,4–6). The aim of this study was to analyze the genotype distribution of the above-mentioned polymorphism in cattle populations of different breeds. The study included a total of 662 individuals of the following breeds: Montbeliard, Limousin, HolsteinFriesian, Angus, Jersey, and Charolais. DNA was extracted from whole peripheral blood, collected from the jugular vein into test tubes containing an anticoagulant (K3EDTA) using the MasterPure DNA Purification Kit for Blood (Epicentre Biotechnology). The primers used for amplification were those described previously (7). PCR amplifications were performed in a 10-μL volume of reaction mixture containing: approximately 60 ng of genomic DNA, 15 pmol of each primer, 1X Taq DNA polymerase buffer with (NH4)2SO4, 1.5 mM MgCl2, 0.2 mM dNTP, 0.4 U of Taq DNA polymerase, and nucleasefree deionized water up to 10 μL. Amplifications were carried out according to the following temperature profile: initial denaturation at 94 °C for 5 min, followed by 35 cycles (denaturation at 94 °C for 30 s, primer annealing at 56 °C for 45 s, extension at 72 °C for 45 s) and final extension at 72 °C for 5 min. PCR products were separated in a 1.5% agarose gel in 1X TBE buffer and stained with ethidium bromide. After electrophoresis, DNA bands were visualized under UV light and photographed. Two alleles (L and S, corresponding to bands of 492 and 408 bp, respectively) and 3 genotypes (LL, LS, and SS) were identified. The analysis included 6 Polish cattle herds consisting of beef (Limousin, Angus, Charolais), dairy (HolsteinFriesian, Jersey), and dual-purpose (Montbeliard) breeds. Allele and genotype frequencies obtained in the present study are given in the Table. There was no polymorphism in the SREBP1 locus in dairy cattle populations. All HolsteinFriesian and Jersey individuals were LL homozygotes. Kaneda et al. (8) also reported monomorphism in Abstract: In the present study, an 84-bp insertion/deletion polymorphism of the sterol regulatory element-binding protein 1 (SREBP1) gene was investigated. The analysis included 6 Polish cattle herds consisting of beef (Limousin, Angus, Charolais), dairy (HolsteinFriesian, Jersey) and dual-purpose (Montbeliard) breeds. The polymorphism was found only in Limousin and Montbeliard animals, in which both the short (S)and long (L)-type alleles occurred. The genotype distribution in Montbeliard cattle was as follows: 0.038, 0.406, and 0.556 for SS, LS, and LL, respectively. To the author’s knowledge, this study was the first to investigate the above-mentioned polymorphic site in the Montbeliard and Jersey cattle populations. As far as SREBP1 function is concerned, the examined locus seems to be a very interesting object for further investigations, such as association studies for fatty acid content and composition in cattle milk.

Polymorphism in dopamine receptor D4 gene is associated with pigeon racing performance

Sequencing and genotyping: Sequencing of PCR products (Table S1) covering the complete exon 3 and parts of introns 2 and 3 from the DRD4 gene revealed four SNPs: the intronic g.129954C>T (ss1751581452), two missense mutations g.129562A>C (p.Arg139Ser; ss1751581453) and g.129456C>T (p.Leu175Phe; ss1751581455) and the synonymous g.129484T>C (ss1751581454). The last one was not further considered. The study involved 123 racing pigeons derived from two unrelated pigeon flocks. SNP g.129562A>C and SNP g.129456C>T were in complete linkage disequilibrium, and only the latter was considered in association analyses. The frequencies of SNPs and of combined genotypes of the DRD4 gene are presented in Table S2.

Nucleotide Substitution in 3' Arm of Bovine MIR-2467 in Five Cattle Breeds

The T > C single nucleotide polymorphism (SNP) in the MIR2467 gene was investigated in order to confirm its presence in cattle genome and to check for possible differences in its genotype distribution among different breeds. Additional purpose of the study was to investigate in silico potential effect of that substitution on the structure and stability of precursor mir-2467. The study involved 634 individuals of five cattle breeds: Angus, Hereford, Holstein-Friesian, Jersey, and Limousin, which were genotyped using PCR-RFLP assay. In this study, the presence of T > C polymorphism at position 24 was observed in all the cattle breeds excepting Hereford. In addition, the differences in the genotype distribution among analyzed breeds were indicated. On the basis of minimum free energy structure prediction, the C allele was indicated to have possible impact on decreasing the stability of the pre-mir-2467, thus altering its ability to regulate target genes expression.

The use of data mining methods for dystocia detection in Polish Holstein-Friesian Black-and-White cattle

Objective The aim of this study was to verify the usefulness of artificial neural networks (ANN), multivariate adaptive regression splines (MARS), naïve Bayes classifier (NBC), general discriminant analysis (GDA), and logistic regression (LR) for dystocia detection in Polish Holstein-Friesian Black-and-White heifers and cows and to indicate the most influential predictors of calving difficulty. Methods A total of 1,342 and 1,699 calving records including six categorical and four continuous predictors were used. Calving category (difficult vs easy or difficult, moderate and easy) was the dependent variable. Results The maximum sensitivity, specificity and accuracy achieved for heifers on the independent test set were 0.855 (for ANN), 0.969 (for NBC), and 0.813 (for GDA), respectively, whereas the values for cows were 0.600 (for ANN), 1.000 and 0.965 (for NBC, GDA, and LR), respectively. With the three categories of calving difficulty, the maximum overall accuracy for heifers and cows was 0.589 (for MARS) and 0.649 (for ANN), respectively. The most influential predictors for heifers were an average calving difficulty score for the dam’s sire, calving age and the mean yield of the farm, where the heifer was kept, whereas for cows, these additionally included: calf sex, the difficulty of the preceding calving, and the mean daily milk yield for the preceding lactation. Conclusion The potential application of the investigated models in dairy cattle farming requires, however, their further improvement in order to reduce the rate of dystocia misdiagnosis and to increase detection reliability.

Cordycepin from Hot Water Extract of Cordyceps militaris Induce Apoptosis in Human Non-Small Lung Carcinoma upon Activation of A3 Adenosine Receptors

Cordyceps militaris hot water extract (CMHW) containing cordycepin (cordycepin-CMHW) was used to study the anti-cancer effects in human A549 non-small cell lung carcinoma cells. Our results showed cordycepin-CMHW can inhibit cell proliferations in A549 cells by activating A3 adenosine receptor (A3AR) via the inactivation of Akt pathways. Cordycepin-CMHW can also induce apoptosis in the A549 cells by enhancing DNA fragmentation and chromatin condensation. We further observed that cordycepin-CMHW up-regulated caspase-9 and increased cleavage of caspase-3 and poly ADP ribose polymerase (PARP) in A549 cells. The results suggested cordycepin-CMHW is a highly selective treatment to de-regulation of the cell proliferation and apoptosis in non-small cell lung carcinoma via signaling pathways generated by A3AR activation.

Identification of Cows Susceptible to Mastitis based on Selected Genotypes by Using Decision Trees and A Generalized Linear Model

Abstract The aim of the present study was to: 1) check whether it would be possible to detect cows susceptible to mastitis at an early stage of their utilization based on selected genotypes and basic production traits in the first three lactations using ensemble data mining methods (boosted classification tress – BT and random forest – RF), 2) find out whether the inclusion of additional production variables for subsequent lactations will improve detection performance of the models, 3) identify the most significant predictors of susceptibility to mastitis, and 4) compare the results obtained by using BT and RF with those for the more traditional generalized linear model (GLZ). A total of 801 records for Polish Holstein-Friesian Black-and-White cows were analyzed. The maximum sensitivity, specificity and accuracy of the test set were 72.13%, 39.73%, 55.90% (BT), 86.89%, 17.81%, 59.49% (RF) and 90.16%, 8.22%, 58.97% (GLZ), respectively. Inclusion of additional variables did not have a significant effect on the model performance. The most significant predictors of susceptibility to mastitis were: milk yield, days in milk, sire’s rank, percentage of Holstein-Friesian genes, whereas calving season and genotypes (lactoferrin, tumor necrosis factor alpha, lysozyme and defensins) were ranked much lower. The applied models (both data mining ones and GLZ) showed low accuracy in detecting cows susceptible to mastitis and therefore some other more discriminating predictors should be used in future research.