Comparison of the Main Constituents in Two Varieties of Proso Millet Using GC-MS.

Proso millet (Panicum miliaceum) is neglected in human nutrition. Thanks to the composition of the grains, millet is suitable for people with celiac disease and it is also useful in the prevention of cardiovascular diseases. For screening the substances in all plant parts of millet via GC-MS, two varieties, Hanacká Mana and Unicum, were used. Substances from the group saccharides, amino acids, fatty acids, carboxylic acids, phytosterols and others were identified in the roots, leaves, stems, and seeds. The highest level of saccharides was found in the stems (83%); amino acids in the roots (6.9%); fatty acids in the seeds (24.6%); carboxylic acids in the roots (3%), phytosterols in the seeds (10.51%); other substances, such as tetramethyl-2-hexadecenol (1.84%) and tocopherols (2.15%), in the leaves; retinal in the roots (1.30%) and squalene in the seeds (1.29%). Saccharides were the dominant group in all plant parts of proso millet followed by fatty acids. The dominant saccharides in all parts of the millet plant were sucrose, fructose and psicose. On the contrary, turanose, trehalose, glucose and cellobiose belonged to the least represented sugars. Additionally, amyrin, miliacin, campesterol, stigmasterol, ß-sitosterol, and others were identified. Varietal variability can be assumed, e.g., in retinal, miliacin or amyrin content.

Different Profiles of Spatial Navigation Deficits In Alzheimer's Disease Biomarker-Positive Versus Biomarker-Negative Older Adults With Amnestic Mild Cognitive Impairment.

Background: Spatial navigation impairment is a promising cognitive marker of Alzheimer's disease (AD) that can reflect the underlying pathology. Objectives: We assessed spatial navigation performance in AD biomarker positive older adults with amnestic mild cognitive impairment (AD aMCI) vs. those AD biomarker negative (non-AD aMCI), and examined associations between navigation performance, MRI measures of brain atrophy, and cerebrospinal fluid (CSF) biomarkers. Methods: A total of 122 participants with AD aMCI (n = 33), non-AD aMCI (n = 31), mild AD dementia (n = 28), and 30 cognitively normal older adults (CN) underwent cognitive assessment, brain MRI (n = 100 had high-quality images for volumetric analysis) and three virtual navigation tasks focused on route learning (body-centered navigation), wayfinding (world-centered navigation) and perspective taking/wayfinding. Cognitively impaired participants underwent CSF biomarker assessment [amyloid-ß1-42, total tau, and phosphorylated tau181 (p-tau181)] and amyloid PET imaging (n = 47 and n = 45, respectively), with a subset having both (n = 19). Results: In route learning, AD aMCI performed worse than non-AD aMCI (p < 0.001), who performed similarly to CN. In wayfinding, aMCI participants performed worse than CN (both p ≤ 0.009) and AD aMCI performed worse than non-AD aMCI in the second task session (p = 0.032). In perspective taking/wayfinding, aMCI participants performed worse than CN (both p ≤ 0.001). AD aMCI and non-AD aMCI did not differ in conventional cognitive tests. Route learning was associated with parietal thickness and amyloid-ß1-42, wayfinding was associated with posterior medial temporal lobe (MTL) volume and p-tau181 and perspective taking/wayfinding was correlated with MRI measures of several brain regions and all CSF biomarkers. Conclusion: AD biomarker positive and negative older adults with aMCI had different profiles of spatial navigation deficits that were associated with posterior MTL and parietal atrophy and reflected AD pathology.

Spatial Pattern Separation Testing Differentiates Alzheimer's Disease Biomarker-Positive and Biomarker-Negative Older Adults With Amnestic Mild Cognitive Impairment.

Background: The hippocampus, entorhinal cortex (EC), and basal forebrain (BF) are among the earliest regions affected by Alzheimer's disease (AD) pathology. They play an essential role in spatial pattern separation, a process critical for accurate discrimination between similar locations. Objective: We examined differences in spatial pattern separation performance between older adults with amnestic mild cognitive impairment (aMCI) with AD versus those with non-Alzheimer's pathologic change (non-AD) and interrelations between volumes of the hippocampal, EC subregions and BF nuclei projecting to these subregions (medial septal nuclei and vertical limb of the diagonal band of Broca - Ch1-2 nuclei) with respect to performance. Methods: Hundred and eighteen older adults were recruited from the Czech Brain Aging Study. Participants with AD aMCI (n = 37), non-AD aMCI (n = 26), mild AD dementia (n = 26), and cognitively normal older adults (CN; n = 29) underwent spatial pattern separation testing, cognitive assessment and brain magnetic resonance imaging. Results: The AD aMCI group had less accurate spatial pattern separation performance than the non-AD aMCI (p = 0.039) and CN (p < 0.001) groups. The AD aMCI and non-AD groups did not differ in other cognitive tests. Decreased BF Ch1-2 volume was indirectly associated with worse performance through reduced hippocampal tail volume and reduced posteromedial EC and hippocampal tail or body volumes operating in serial. Conclusion: The study demonstrates that spatial pattern separation testing differentiates AD biomarker positive and negative older adults with aMCI and provides evidence that BF Ch1-2 nuclei influence spatial pattern separation through the posteromedial EC and the posterior hippocampus.

Spatial Navigation and Visuospatial Strategies in Typical and Atypical Aging.

Age-related spatial navigation decline is more pronounced in patients with mild cognitive impairment (MCI) and Alzheimer's disease (AD) dementia. We used a realistic-looking virtual navigation test suite to analyze different aspects of visuospatial processing in typical and atypical aging. A total of 219 older adults were recruited from the Czech Brain Aging Study cohort. Cognitively normal older adults (CN; n = 78), patients with amnestic MCI (n = 75), and those with mild AD dementia (n = 66) underwent three navigational tasks, cognitive assessment, and brain MRI. Route learning and wayfinding/perspective-taking tasks distinguished the groups as performance and learning declined and specific visuospatial strategies were less utilized with increasing cognitive impairment. Increased perspective shift and utilization of non-specific strategies were associated with worse task performance across the groups. Primacy and recency effects were observed across the groups in the route learning and the wayfinding/perspective-taking task, respectively. In addition, a primacy effect was present in the wayfinding/perspective-taking task in the CN older adults. More effective spatial navigation was associated with better memory and executive functions. The results demonstrate that a realistic and ecologically valid spatial navigation test suite can reveal different aspects of visuospatial processing in typical and atypical aging.

Spatial Pattern Separation in Early Alzheimer's Disease.

BACKGROUND: The hippocampus, entorhinal cortex, and basal forebrain are among the first brain structures affected by Alzheimer's disease (AD). They play an essential role in spatial pattern separation, a process critical for accurate encoding of similar spatial information. OBJECTIVE: Our aim was to examine spatial pattern separation and its association with volumetric changes of the hippocampus, entorhinal cortex, and basal forebrain nuclei projecting to the hippocampus (the medial septal nuclei and vertical limb of the diagonal band of Broca - Ch1-2 nuclei) in the biomarker-defined early clinical stages of AD. METHODS: A total of 98 older adults were recruited from the Czech Brain Aging Study cohort. The participants with amnestic mild cognitive impairment (aMCI) due to AD (n = 44), mild AD dementia (n = 31), and cognitively normal older adults (CN; n = 23) underwent spatial pattern separation testing, comprehensive cognitive assessment, and MRI brain volumetry. RESULTS: Spatial pattern separation accuracy was lower in the early clinical stages of AD compared to the CN group (p < 0.001) and decreased with disease severity (CN > aMCI due to AD > AD dementia). Controlling for general memory and cognitive performance, demographic characteristics and psychological factors did not change the results. Hippocampal and Ch1-2 volumes were directly associated with spatial pattern separation performance while the entorhinal cortex operated on pattern separation indirectly through the hippocampus. CONCLUSION: Smaller volumes of the hippocampus, entorhinal cortex, and basal forebrain Ch1-2 nuclei are linked to spatial pattern separation impairment in biomarker-defined early clinical AD and may contribute to AD-related spatial memory deficits.

The first report of animal genotypes of Cryptosporidium parvum in immunosuppressed and immunocompetent humans in Slovakia.

The aim of our study was to determine species and genotypes of Cryptosporidium in patients suffering from immunosuppressive illnesses, but also in immunocompetent patients suffering from diarrhoea. A total of 80 samples of faeces were collected from both immunosuppressed and immunocompetent patients. The immunosuppressed patients (65 samples) - 35 adult patients (group A) and 30 children (group B) were hospitalized at the Clinic of Oncohemathology. Samples from immunocompetent humans (15 samples, group C) were taken from patients with clinical signs of acute diarrhoea. With the use of molecular methods targeting the 60 kDa glycoprotein (GP60) gene region, we have identified multiple genotypes of Cryptosporidium. parvum and Cryptosporidium. hominis in immunocompromised, but also in immunocompetent individuals (C. hominis IbA10G2, IeA12G3T3; C. parvum IIaA10G1R1, IIaA11G2R1, IIaA12G2R1, IIaA13G1R1, IIaA14G1R1, IIaA14G2R1, IIaA17G1R1 and IIaA18G1R1). This is the first report of the occurrence of genotypes IIaA10G1R1, IIa12G2R1 and IIaA18G1R1 in human hosts.

Contribution to the study of rutin stability in the achenes of Tartary buckwheat (Fagopyrum tataricum).

Tartary buckwheat achenes are rich in rutin. However, its utilisation as a food is low due to the bitter taste of the products because of the transformation of rutin to quercetin. The aim of this study was to determine the stability of rutin and selected flavonoids, which are influenced by the different sample preparations exposing the achenes to higher temperatures (60 °C, 100 °C, and 140 °C), and also to determine the specificity of the rutin degrading enzymes in Tartary buckwheat. The achenes were directly extracted with methanol pre-extracted with hexane, or lyophilised before extraction. The rutin and quercetin contents were determined by HPLC. The rutin-degrading enzymes in Tartary buckwheat achenes were not inactivated after a heat treatment up to 140 °C acting on them for three hours; neither to lyophilisation nor pre-extraction with hexane. The rutin-degrading enzymes probably caused the deglycosylation of kaempferol-3-O-rutinoside, kaempferol-3-O-ß-d-glucoside and piceid. Rutin-degrading enzymes were inactivated with 100% methanol.

Uptake of caprolactam and its influence on growth and oxygen production of Desmodesmus quadricauda algae.

The consumption of polyamides produced from caprolactam is increasing continuously, and for that reason the danger of environmental contamination by this lactam is also rising. This study's aim was to evaluate the influence of caprolactam on the growth and oxygen production of the green alga Desmodesmus quadricauda and on caprolactam uptake by this alga. The presence of caprolactam in water was observed to cause the algae significantly to increase its oxygen production. Caprolactam concentration of 5,000 mg/L stopped algae growth after 6 days and influenced coenobia structure (seen as disappearance of pyrenoids, deformation of cells) but did not decrease the number of cells in the coenobia. Caprolactam uptake is probably passive but relatively rapid. Maximum concentration in the algae was reached after 18-24 h.

Molecular characterization and first report of Cryptosporidium genotypes in human population in the Slovak Republic.

In our study, we examined 91 fecal samples from five different groups of people containing HIV patients, hemodialysis patients, kidney transplant recipients, immunocompetent humans without clinical signs, and humans with suspected cryptosporidiosis. The purpose of our study was to determine species and genotype composition of representatives of Cryptosporidium spp. using PCR analysis of small subunit ribosomal RNA gene and 60-kDa glycoprotein gene and examine their phylogenetic relationship. In HIV-positive/AIDS-infected group of patients and in hemodialysis patients, no presence of Cryptosporidium species was detected. In two kidney transplant recipients, we detected species/genotypes Cryptosporidium parvum IIaA13G1T1R1 (KT355488) and Cryptosporidium hominis IaA11G2R8 (KT355489) and in two immunocompetent patients with clinical symptoms, we identified Cryptosporidium muris and C. hominis IbA10G2T1 (KT355490). In the group of healthy immunocompetent individuals without clinical signs, we identified species/genotype C. hominis IbA11G2 (KT355491) in one sample.

Verification of presence of caprolactam in sprouted achenes of Fagopyrum esculentum Moench and its influence on plant phenolic compound content.

The presence of caprolactam, a precursor of Nylon-6, among those synthetic polymers which are widely-spread throughout the environment, could be the reason for its being found in plants. The aim of this work was to confirm the previously described presence of caprolactam in dry and sprouted achenes, as well as in achene exudates of common buckwheat (Fagopyrum esculentum Moench). When the lyophilized sprouted and dry buckwheat achenes, along with exudates from growth experiments, with caprolactam-free medium were analysed by HPLC, no caprolactam was found. After addition of caprolactam into the growth medium, we confirmed the uptake of caprolactam in the lyophilized sprouted buckwheat achenes. The uptake of caprolactam is also a function of light conditions during the growth experiments. Caprolactam also inhibits the content of phenolic compounds; especially rutin, vitexin, isovitexin, orientin, and homoorientin in buckwheat plants.

Cryptosporidium scrofarum n. sp. (Apicomplexa: Cryptosporidiidae) in domestic pigs (Sus scrofa).

We describe the morphological, biological, and molecular characteristics of Cryptosporidium pig genotype II and propose the species name Cryptosporidium scrofarum n. sp. to reflect its prevalence in adult pigs worldwide. Oocysts of C. scrofarum are morphologically indistinguishable from C. parvum, measuring 4.81-5.96 µm (mean=5.16)×4.23-5.29 µm (mean=4.83) with a length to width ratio of 1.07±0.06 (n=400). Oocysts of C. scrofarum obtained from a naturally infected pig were infectious for 8-week-old pigs but not 4-week-old pigs. The prepatent period in 8-week-old Cryptosporidium-naive pigs was 4-6 days and the patent period was longer than 30 days. The infection intensity of C. scrofarum in pigs was generally low, in the range 250-4000 oocysts per gram of feces. Infected pigs showed no clinical signs of cryptosporidiosis and no pathology was detected. Cryptosporidium scrofarum was not infectious for adult SCID mice, adult BALB/c mice, Mongolian gerbils (Meriones unguiculatus), southern multimammate mice (Mastomys coucha), yellow-necked mice (Apodemus flavicollis), or guinea pigs (Cavia porcellus). Phylogenetic analyses based on small subunit rRNA, actin, and heat shock protein 70 gene sequences revealed that C. scrofarum is genetically distinct from all known Cryptosporidium species.

The influence of organic and conventional crop management, variety and year on the yield and flavonoid level in common buckwheat groats.

Common buckwheat (Fagopyrum esculentum Moench) is a pseudocereal that is cultivated for its achenes that possess a high level of flavonoids. Some scientific studies have found that the growing conditions can significantly affect the flavonoid content in conventional and organic food products. Hence, the present study compared conventionally and organically produced common buckwheat with respect to the content of selected flavonoids, thousand achenes weight and achene yield. Three buckwheat varieties were grown with organic and conventional crop management under the same environmental conditions. The thousand achenes weight and achene yield were not significantly influenced by way of crop management. The level of rutin, epicatechin, catechin, and epicatechin gallate in buckwheat groats (hulled achenes) were quantified with high pressure liquid chromatography. Only rutin and epicatechin gallate reached significantly higher level in organic groats. However, the differences were influenced by environmental conditions in the given year and variety.

Determination of quercetin glycosides and free quercetin in buckwheat by capillary micellar electrokinetic chromatography.

A simple and reliable method for determination of quercetin glycosides and free quercetin in buckwheat flower, leaves, stems and achenes was developed. The method consists of flavonoid extraction from freeze-dried homogenous material in 50% v/v methanol solution and in presence of an antioxidant, cleaning of extract and analyte isolation using SPE. Analytical step uses capillary micellar electrokinetic chromatography. The working ranges, LOD and LOQ, recovery, precision and measurement uncertainty were calculated. The method is suitable for samples from buckwheat. The highest content of rutin was found in flowers of both kinds of buckwheat (99,400 mg/kg in F. esculentum, 108,000 mg/kg in F. tataricum). The free quercetin occurs in flowers and achenes of F. esculentum, whereas flowers and achenes of F. tataricum contained quercitrin.

A new epigenetic marker: the replication-coupled, cell cycle-dependent, dual modification of the histone H4 tail.

Recently, we described the cold-dependent detection of an epitope, epiC, that was selectively recognized by a monoclonal anti-actin antibody at 4 degrees C, but not at RT, in the early replicating chromatin domains of human fibroblast cell nuclei and chromosomes. EpiC was present in a distinct cell cycle window extending from S-phase throughout mitosis until early G1-phase of the next cell generation, indicating its possible involvement in the transfer/maintenance of epigenetic information on transcriptionally competent parts of the genome. However, the molecular nature of epiC remained unresolved. Here we identified epiC as a dual post-translational modification on the same histone H4 tail, which was immunodetected for the first time. We show that the antibody selectively recognized a synthetic peptide of the histone H4 region K12-L22 containing acetylated K16 and dimethylated K20 (H4K16ac-K20me2) at 4 degrees C, but not at RT. Moreover, we show that the peptide containing acetylated K16 and either unmodified or monomethylated K20 was recognized by this antibody at both temperatures. The present and previous results together indicate that, by acetylation of histone H4 K16 during S-phase, the early replicating chromatin domains acquire the H4K16ac-K20me2 epigenetic label that persists on the chromatin throughout mitosis and become deacetylated during early G1-phase of the next cell cycle.

Level of catechin, myricetin, quercetin and isoquercitrin in buckwheat (Fagopyrum esculentum Moench), changes of their levels during vegetation and their effect on the growth of selected weeds.

Buckwheat is well-known as a crop rich in flavonoids, however, attention has usually only been paid to the main flavonoid rutin as an important natural antioxidant or as a possible allelopathic compound. Therefore, some of the other constituents found within individual plant parts of buckwheat (isoquercitrin, quercetin, catechin, and myricetin), as well as changes of their level during the growing season, were determined by HPLC analysis. The effects of these compounds on plant growth were proved on seven plant species. In buckwheat, isoquercitrin represented the largest component of the selected compounds. The strongest inhibitive effects on the growth of those selected plants were produced by catechin. Quercetin and isoquercitrin had weak inhibitive effects. Myricetin did not show any influence on plant growth. Hence we suppose that myricetin, isoquercetin and quercetin do not have important function in allelopathy of buckwheat. Buckwheat as row material for functional foods could be a significant source of another antioxidant, isoquercitrin.

Rutin and total quercetin content in amaranth (Amaranthus spp.).

The aim of the study was to confirm the presence of rutin, one of the most common quercetin glycosides, and other quercetin derivatives in plants of genus Amaranthus, to investigate the influence of the species and variety on rutin distribution in the plant and content changes during growing season. The rutin content was determined by micellar electrokinetic capillary chromatography in individual plant parts at the beginning of the growth, at the flowering stage and at the maturity stage of five Amaranthus species. The total quercetin content was determined by micellar electrokinetic capillary chromatography too. The rutin content in amaranth ranged from 0.08 (in seeds) to 24.5 g/kg dry matter (in leaves). Comparison of the determined total quercetin content and the calculated content of quercetin released from rutin did not prove important presence of quercetin or other quercetin derivatives than rutin. Only amaranth leaves sampled at the maturity stage probably contained quercetin or quercetin derivatives. Significant differences in the rutin content were established among species and as well varieties. Amaranthus hybrid and A. cruentus were the best sources of rutin.

Exudation of allelopathic substances in buckwheat (Fagopyrum esculentum Moench).

Root exudates of the common buckwheat, especially phenolic compounds, were studied. Their contents, both in the soil during the growing season and in agar medium during germination, were determined by HPLC and GC-MS. The allelopathic activity of the soil from a buckwheat stand was evaluated, as well. Palmitic acid, squalene, epicatechin, vitexin, a gallic acid derivative, and a quercetin derivative were the main compounds of the agar medium. In the soil, palmitic acid methyl ester, vanillic acid, rutin, a gallic acid derivative, and a 4-hydroxyacetophenone derivative were identified. The effects of vitexin, squalene, epicatechin, 4-hydroxyacetophenone, and vanillic and gallic acids were tested on eight plant species. Inhibitive effects were observed in the cases of 4-hydroxyacetophenone and vanillic and gallic acids. Comparisons of the identified compounds and inhibitive effects of soil extracts indicated that palmitic acid and the gallic acid derivative probably have an important function in the allelopathic root response of buckwheat.

Distribution of Vitamin E, squalene, epicatechin, and rutin in common buckwheat plants (Fagopyrum esculentum Moench).

Buckwheat leaves and young parts of the plant are consumed in some countries as a vegetable. Green flour, obtained by milling of the dried plants, is used as a natural food colorant. The distribution of vitamin E, squalene, epicatechin, and rutin (as the most important antioxidants) within buckwheat plants, as well as changes of their content within leaves during the growing season, were determined by GC-MS and HPLC analyses. alpha-Tocopherol was found as the main component of vitamin E in all parts of the plant; epicatechin and squalene were also detected. For the use of buckwheat as an antioxidant source in the human diet, the most suitable part of the plants seems to be the leaves and the flowers at the stage of full flowering due to the considerable amounts of rutin and epicatechin. alpha-Tocopherol content correlates positively with temperature, drought, and duration of solar radiation. Certain differences appear among varieties of buckwheat, especially in their squalene and rutin contents.

Content and quality of protein in proso millet (Panicum miliaceum L.) varieties.

Proso millet belongs to the oldest cereals that human is using. Eight varieties of proso millet were cultivated in Ceske Budejovice from 1998 to 2000 and Cerveny Dvur from 1999 to 2000. The crude protein content was determined according to Kjehladl method and amino acid content was determined chromatographically after acid and oxidative acid hydrolysis. Although the protein content of proso (11.6% of dry matter) was similar to wheat, the grain of proso was significant richer in essential amino acids (leucine, isoleucine, methionine) then wheat. Hence, the protein quality of proso (Essential Amino Acid Index) was higher (51%) compared to wheat. The proso grain contained about 3.3 g kg(-1) of the limiting amino acid-lysine. Significant differences in protein and its quality were found among the evaluated proso varieties. The varieties Toldanskoe and Lipetskoe were the most different from the others in protein and amino acid content and Amino Acid Score of individual acids. They had the lowest content and quality of protein. The seed coat of these varieties was red. The amino acid and protein content was significantly influenced by weather during the year. Dry conditions caused an increase of protein but its quality was decreased.