Caffeoylquinic Acids,Cytotoxic, Antioxidant,Acetylcholinesterase and Tyrosinase Enzyme Inhibitory Activities of Six Inula Species from Bulgaria

Chlorogenic (5‐CQA), 1,5‐, 3,5‐, 4,5‐ and 3,4‐dicaffeoylquinic (DCQA) acids were identified and quantified in the methanol extracts of Inula oculus‐christi L., I. bifrons L., I. aschersoniana Janka var. aschersoniana, I. ensifolia L., I. conyza (Griess .) DC. and I. germanica L. by HPLC analysis. The amount of 5‐CQA varied from 5.48 to 28.44 mg/g DE and the highest content was detected in I. ensifolia. 1,5‐DCQA (4.05–55.25 mg/g DE) was the most abundant dicaffeoyl ester of quinic acid followed by 3,5‐DCQA, 4,5‐DCQA and 3,4‐DCQA. The extract of I. ensifolia showed the highest total phenolic content (119.92±0.95 mg GAE/g DE) and exhibited the strongest DPPH radical scavenging activity (69.41±0.55 %). I. bifrons extract was found to be the most active sample against ABTS^.+ (TEAC 0.257±0.012 mg/mL) and the best tyrosinase inhibitor. The studied extracts demonstrated a low inhibitory effect towards acetylcholinesterase and possessed low cytotoxicity in concentration range from 10 to 300 μg/mL toward non‐cancer (MDCK II) and cancer (A 549) cells.     

Micropropagation of the endangered species Malus niedzwetzkyana for conservation biodiversity in Kazakhstan

In Vitro Cellular & Developmental Biology - Plant - Biodiversity conservation requires advanced and effective ex situ plant propagation techniques. The present study was conducted to optimize...     

Safflower (Carthamus Tinctorius L.) a Potential Source of Drugs against Cryptococcal Infections,Malaria and Leishmaniasis

In this research we present that Carthamus Tinctorius L. (gen. Asteraceae, otherwise known as Safflower) (Fig. 1) may contain agents active in Cryptococcal infections, malaria and Leishmaniasis, as treatment options are becoming scarce due to drug resistance development. Phytochemistry and pharmacological activities (antimicrobial, antimalarial, antileishmanial) of C. tinctorius L. were analyzed. The composition of volatile oil of safflower dried flowers was analyzed by gas chromatography-mass spectrophotometry with flame ionization detector (GC-FID) and in vitro sensitivity assays were performed to assess biological activity. 8 known and 3 unknown compounds were detected in the extract (Fig. 1). Then the Safflower ointment was manufactured and its acute toxicity study on rats was tested. The volatile oil of C. tinctorius L exhibited activity against Cryptococcus neoformans, Plasmodium falciparum and Leishmania donovani. Safflower volatile oil has anticryptococcal, antimalarial and antileishmanial effects. The prepared ointment had an excellent acute toxicity safety profile.     

Chemical diversity of volatiles of Teucrium orientale L. var. orientale, var. puberulens, and var. glabrescens determined by simultaneous GC-FID and GC/MS techniques

In the present work, three varieties of Teucrium orientale, var. orientale, var. puberulens, and var. glabrescens, were collected and investigated for chemical composition of the oils. Subsequent gas chromatography (GC-FID) and gas chromatography coupled to mass spectrometry (GC/MS) revealed high abundance of sesquiterpenes in the essential oils analyzed. All the oils contained β-caryophyllene (22.6, 8.5, and 6.3%, resp.) and hexadecanoic acid (7.9, 12.8, and 13.1%). Germacrene D (24.6 and 33.4%) and bicyclogermacrene (6.7 and 8.5%) were found to be the main constituents of var. orientale and var. puberulens, respectively. The high percentages of β-cubebene (26.9%), α-cubebene (9.0%), and α-copaene (7.2%) established the diversity of var. glabrescens. The qualitative difference between the essential oils allowed the differentiation between the varieties in agreement with the morphological observations described in Flora of Turkey for each variety studied. In addition, a cluster analysis of twelve Teucrium taxa based on the essential-oil composition has been carried out. Hovewer, the analysis did not clearly reflect the infrageneric classification of the genus, it largely confirmed the relationships between the infraspecific taxa of Teucrium orientale and T. chamaedrys.     

Advantages of Amplifluor-like SNP markers over KASP in plant genotyping

Background

KASP (KBioscience Competitive Allele Specific PCR) and Amplifluor (Amplification with fluorescence) SNP markers are two prominent technologies based upon a shared identical Allele-specific PCR platform.

Methods

Amplifluor-like SNP and KASP analysis was carried out using published and own design of Universal probes (UPs) and Gene-specific primers (GSPs).

Results

Advantages of the Amplifluor-like system over KASP include the significantly lower costs and much greater flexibility in the adjustment and development of ‘self-designed’ dual fluorescently-labelled UPs and regular GSPs. The presented results include optimisation of ‘tail’ length in UPs and GSPs, protocol adjustment, and the use of various fluorophores in different qPCR instruments. The compatibility of the KASP Master-mix in both original and Amplifluor-like systems has been demonstrated in the presented results, proving their similar principles. Results of SNP scoring with rare alleles in addition to more frequently occurring alleles are shown.

Conclusions

The Amplifluor-like system produces SNP genotyping results with a level of sensitivity and accuracy comparable to KASP but at a significantly cheaper cost and with much greater flexibility for UPs with self-designed GSPs.

     

Differences in photosynthetic activity, chlorophyll and carotenoid levels, and in chlorophyll fluorescence parameters in green sun and shade leaves of Ginkgo and Fagus

The differences in pigment levels and photosynthetic activity of green sun and shade leaves of ginkgo (Ginkgo biloba L.) and beech (Fagus sylvatica L.) are described. Sun leaves of both tree species possessed higher levels in chlorophylls (Chl) and carotenoids on a leaf area basis, higher values for the ratio Chl a/b and lower values for the ratio Chl/carotenoids (a+b)/(x+c) in comparison to shade leaves. The higher photosynthetic rates P(N) of sun leaves (ginkgo 5.4+/-0.9 and beech 8.5+/-2.1 micromol m(-2)s(-1)) were also reflected by higher values for the Chl fluorescence decrease ratios R(F)(d) 690 and R(F)(d) 735. In contrast, the shade leaves had lower P(N) rates (ginkgo 2.4+/-0.3 and beech 1.8+/-1.2 micromol m(-2)s(-1)). In both tree species the stomatal conductance G(s) was significantly higher in sun (range: 70-19 1 mmol m(-2)s(-1)) as compared to shade leaves (range: 5-55 mmol m(-2)s(-1)). In fact, at saturating light conditions there existed a close correlation between G(s) values and P(N) rates. Differences between sun and shade leaves also existed in several other Chl fluorescence ratios (F(v)/F(m), F(v)/F(o), and the stress adaptation index Ap). The results clearly demonstrate that the fan-shaped gymnosperm ginkgo leaves show the same high and low irradiance adaptation response as the angiosperm beech leaves.     

A new strain group of common carp: The genetic differences and admixture events between Cyprinus carpio breeds

Common carp (Cyprinus carpio) has an outstanding economic importance in freshwater aquaculture due to its high adaptive capacity to both food and environment. In fact, it is the third most farmed fish species worldwide according to the Food and Agriculture Organization. More than four million tons of common carp are produced annually in aquaculture, and more than a hundred thousand tons are caught from the wild. Historically, the common carp was also the first fish species to be domesticated in ancient China, and now, there is a huge variety of domestic carp strains worldwide. In the present study, we used double digestion restriction site‐associated DNA sequencing to genotype several European common carp strains and showed that they are divided into two distinct groups. One of them includes central European common carp strains as well as Ponto–Caspian wild common carp populations, whereas the other group contains several common carp strains that originated in the Soviet Union, mostly as cold‐resistant strains. We believe that breeding with wild Amur carp and subsequent selection of the hybrids for resistance to adverse environmental conditions was the attribute of the second group. We assessed the contribution of wild Amur carp inheritance to the common carp strains and discovered discriminating genes, which differed in allele frequencies between groups. Taken together, our results improve our current understanding of the genetic variability of common carp, namely the structure of natural and artificial carp populations, and the contribution of wild carp traits to domestic strains.     

Chemical Diversity of Volatiles of Teucrium orientale L. var. orientale,var. puberulens,and var. glabrescens Determined by Simultaneous GC‐FID and GC/MS Techniques

In the present work, three varieties of Teucrium orientale, var. orientale, var. puberulens, and var. glabrescens, were collected and investigated for chemical composition of the oils. Subsequent gas chromatography (GC‐FID) and gas chromatography coupled to mass spectrometry (GC/MS) revealed high abundance of sesquiterpenes in the essential oils analyzed. All the oils contained β‐caryophyllene (22.6, 8.5, and 6.3%, resp.) and hexadecanoic acid (7.9, 12.8, and 13.1%). Germacrene D (24.6 and 33.4%) and bicyclogermacrene (6.7 and 8.5%) were found to be the main constituents of var. orientale and var. puberulens, respectively. The high percentages of β‐cubebene (26.9%), α‐cubebene (9.0%), and α‐copaene (7.2%) established the diversity of var. glabrescens. The qualitative difference between the essential oils allowed the differentiation between the varieties in agreement with the morphological observations described in Flora of Turkey for each variety studied. In addition, a cluster analysis of twelve Teucrium taxa based on the essential‐oil composition has been carried out. Hovewer, the analysis did not clearly reflect the infrageneric classification of the genus, it largely confirmed the relationships between the infraspecific taxa of Teucrium orientale and T. chamaedrys.     

Genetic risk factors for restenosis after percutaneous coronary intervention in Kazakh population

Background

After coronary stenting, the risk of developing restenosis is from 20 to 35 %. The aim of the present study is to investigate the association of genetic variation in candidate genes in patients diagnosed with restenosis in the Kazakh population.

Methods

Four hundred fifty-nine patients were recruited to the study; 91 patients were also diagnosed with diabetes and were excluded from the sampling. DNA was extracted with the salting-out method. The patients were genotyped for 53 single-nucleotide polymorphisms. Genotyping was performed on the QuantStudio 12K Flex (Life Technologies). Differences in distribution of BMI score among different genotype groups were compared by analysis of variance (ANOVA). Also, statistical analysis was performed using R and PLINK v.1.07. Haplotype frequencies and LD measures were estimated by using the software Haploview 4.2.

Results

A logistic regression analysis found a significant difference in restenosis rates for different genotypes. FGB (rs1800790) is significantly associated with restenosis after stenting (OR?=?2.924, P?=?2.3E?06, additive model) in the Kazakh population. CD14 (rs2569190) showed a significant association in the additive (OR?=?0.08033, P?=?2.11E?09) and dominant models (OR?=?0.05359, P?=?4.15E?11). NOS3 (rs1799983) was also highly associated with development of restenosis after stenting in additive (OR?=?20.05, P?=?2.74 E?12) and recessive models (OR?=?22.24, P?=?6.811E?10).

Conclusions

Our results indicate that FGB (rs1800790), CD14 (rs2569190), and NOS3 (rs1799983) SNPs could be genetic markers for development of restenosis in Kazakh population. Adjustment for potential confounder factor BMI gave almost the same results.

     

Furanocoumarin Content,Antioxidant Activity,and Inhibitory Potential of Heracleum verticillatum,Heracleum sibiricum,Heracleum angustisectum,and Heracleum ternatum Extracts against Enzymes Involved in Alzheimer's Disease and Type II Diabetes

Hexane extracts of Heracleum verticillatum, H. sibiricum, H. angustisectum, and H. ternatum were studied for their furanocoumarin content antioxidant potential and acetylcholinesterase and α‐amylase inhibitory activities. Quantification of the furanocoumarins was performed by ¹H‐NMR. Pimpinellin was found to be the main component in the roots of all studied species. Bergapten and imperatorin were the major compounds in the fruits of H. sibiricum and H. verticillatum, respectively, while byakangelicol dominated in H. angustisectum and H. ternatum fruits. The leaf and fruit extracts of H. angustisectum demonstrated the highest DPPH radical scavenging activity and TEAC (IC₅₀ 0.58 mg/mL and 1.83 mm , respectively). The root extracts of H. verticillatum and H. angustisectum were found to be the most effective against acetylcholinesterase (IC₅₀ 0.30 and 0.34 mg/mL, respectively). The studied extracts were not active or demonstrated a weak inhibitory effect (%Inh. up to 29.7) towards α‐amylase.