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1.
Food Res Int ; 111: 244-255, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-30007683

RESUMO

Alginate hydrogels are often used for immobilization of plant-derived bioactive compounds by fast and simple ionic gelation technique. However, the structure of alginate gel network is very porous and mostly result with high-diffusion rates of encapsulated compound, what limits its application as delivery vehicle. In order to prevent losses of bioactives and prepare efficient encapsulation systems, the aim of this study was to evaluate a potential of new natural fillers, cocoa powder (CP) and carob (C) for structuring alginate network aimed for encapsulation of aqueous dandelion (Taraxacum officinale L.) leaf extract using ionic gelation. Whey protein isolates served as a standard filler. The influence of different concentrations of gelling medium (2% and 3% calcium chloride) on encapsulation properties of alginate systems was also evaluated. Calcium concentration affected morphological properties (more acceptable when using 3% CaCl2), while textural properties and encapsulation efficiency of polyphenols and retained antioxidant capacity were more influenced by selected delivery materials. Alginate-whey protein isolates beads were scored with the highest loading capacity of polyphenols (>93%), while newly formulated binary mixtures (alginate-cocoa powder and alginate-carob) also enabled highly efficient entrapment of polyphenols (>88%). The slowest release of polyphenols in simulated gastrointestinal fluids were obtained when alginate was combined with CP and C, where system alginate-cocoa powder prepared with lower concentration of calcium chloride (2% CaCl2) enabled the most extended release of total polyphenols and hydroxycinnamic acids. Obtained results strongly justified implementation of new plant-derived functional fillers (cocoa powder and carob) for encapsulation purposes and opened new directions for designing of binary carrier's.


Assuntos
Alginatos/química , Cacau/química , Galactanos/química , Mananas/química , Extratos Vegetais/química , Gomas Vegetais/química , Polifenóis/química , Taraxacum/química , Alginatos/metabolismo , Cacau/metabolismo , Cloreto de Cálcio/química , Cloreto de Cálcio/metabolismo , Galactanos/metabolismo , Géis , Hidrogéis , Mananas/metabolismo , Extratos Vegetais/metabolismo , Gomas Vegetais/metabolismo , Polifenóis/metabolismo , Taraxacum/metabolismo
2.
Molecules ; 23(6)2018 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-29865166

RESUMO

There is growing interest regarding the use of herbal preparations based on Cannabis sativa for medicinal purposes, despite the poorly understood interactions of their main constituent Δ8-tetrahydrocannabinol (THC) with conventional drugs, especially cytostatics. The objective of this pilot study was to prove whether the concomitant intake of THC impaired liver function in male Wistar rats treated with the anticancer drug irinotecan (IRI), and evaluate the toxic effects associated with this exposure. IRI was administered once intraperitoneally (at 100 mg/kg of the body weight (b.w.)), while THC was administered per os repeatedly for 1, 3, and 7 days (at 7 mg/kg b.w.). Functional liver impairments were studied using biochemical markers of liver function (aspartate aminotransferase-AST, alanine aminotransferase-ALP, alkaline phosphatase-AP, and bilirubin) in rats given a combined treatment, single IRI, single THC, and control groups. Using common oxidative stress biomarkers, along with measurement of primary DNA damage in hepatocytes, the degree of impairments caused at the cellular level was also evaluated. THC caused a time-dependent enhancement of acute toxicity in IRI-treated rats, which was confirmed by body and liver weight reduction. Although single THC affected ALP and AP levels more than single IRI, the levels of liver function markers measured after the administration of a combined treatment mostly did not significantly differ from control. Combined exposure led to increased oxidative stress responses in 3- and 7-day treatments, compared to single IRI. Single IRI caused the highest DNA damage at all timepoints. Continuous 7-day oral exposure to single THC caused an increased mean value of comet tail length compared to its shorter treatments. Concomitant intake of THC slightly affected the levels of IRI genotoxicity at all timepoints, but not in a consistent manner. Further studies are needed to prove our preliminary observations, clarify the underlying mechanisms behind IRI and THC interactions, and unambiguously confirm or reject the assumptions made herein.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Camptotecina/análogos & derivados , Dronabinol/farmacologia , Fígado/efeitos dos fármacos , Administração Oral , Animais , Biomarcadores/metabolismo , Peso Corporal/efeitos dos fármacos , Camptotecina/farmacologia , Dano ao DNA , Relação Dose-Resposta a Droga , Irinotecano , Fígado/fisiopatologia , Testes de Função Hepática , Masculino , Testes de Mutagenicidade , Tamanho do Órgão/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Ratos Wistar
3.
Forensic Sci Int Genet ; 34: 249-256, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29573605

RESUMO

In order to implement X-chromosome short tandem repeat (X-STR) typing into routine forensic practice, reference database of a given population should be established. Therefore we extended already published data with additional 397 blood samples from unrelated Croatian citizens, and analyzed the total of 995 samples (549 male and 446 female) typed by Investigator® Argus X-12 Kit. To test genetic homogeneity of consecutively processed five historic-cultural regions covering the entire national territory, we calculated pairwise Fst genetic distances between regions based on allele and full haplotype frequencies. Since the comparison did not yield any statistically significant difference, we integrated STR profile information from all regions and used the whole data set to calculate forensic parameters. The most informative marker is DXS10135 (polymorphism information content (PIC = 0.929) and the most informative linkage group (LG) is LG1 (PIC = 0.996). We confirmed linkage disequilibrium (LD) for seven marker pairs belonging to LG2, LG3 and LG4. By including LD information, we calculated cumulative power of discrimination that amounted to 0.999999999997 in females and 0.999999005 in males. We also compared Croatia with 13 European populations based on haplotype frequencies and detected no statistically significant Fst values after Bonferroni correction in any LG. Multi-dimensional scaling plot revealed tight grouping of four Croatian regions amongst populations of southern, central and northern Europe, with the exception of northern Croatia. In this study we gave the first extensive overview of aberrant profiles encountered during Investigator® Argus X-12 typing. We found ten profiles consistent with single locus duplication followed by tetranucleotide tract length polymorphism. Locus DXS10079 is by far the most frequently affected one, presumably mutated in eight samples. We also found four profiles consistent with X-chromosome aneuploidy (three profiles with XXX pattern and one profile with XXY pattern). In conclusion, we established integral forensic Croatian X-chromosome database, proved forensic pertinence of Investigator® Argus X-12 Kit for the entire Croatian population and identified locus DXS10079 as a potential duplication hotspot.


Assuntos
Cromossomos Humanos X , Bases de Dados de Ácidos Nucleicos , Genética Populacional , Repetições de Microssatélites , Croácia , Impressões Digitais de DNA , Feminino , Loci Gênicos , Humanos , Desequilíbrio de Ligação , Masculino , Polimorfismo Genético
4.
J Phycol ; 54(2): 275-298, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29419886

RESUMO

The diatom genus Entomoneis is known from the benthos and plankton of marine, brackish, and freshwaters. Entomoneis includes diatoms with a bilobate keel elevated above the valve surface, a sigmoid canal raphe, and numerous girdle bands. Owing mostly to the scarcity of molecular data for a diverse set of species, the phylogeny of Entomoneis has not been investigated in depth. The few previous studies that included Entomoneis were focused on broader questions and the available data were from a small number of either unidentified Entomoneis or well-known species (e.g., E. paludosa). Since the first description of new species combining both molecular and morphological characters (E. tenera), we have continued to cultivate and investigate Entomoneis in the plankton of the Adriatic Sea. Combined multigene phylogeny (SSU rDNA sequences, rbcL, and psbC genes) and morphological observations (LM, SEM and TEM) revealed six new Entomoneis species supported by phylogenetic and morphological data: E. pusilla, E. gracilis, E. vilicicii, E. infula, E. adriatica, and E. umbratica. The most important morphological features for species delineation were cell shape, the degree and mode of torsion, valve apices, the appearance and structure of the transition between keel and valve body, the ultrastructure and the shape of the girdle bands, and the arrangement and density of perforations along the valve and valvocopulae. Our results highlight the underappreciated diversity of Entomoneis and call for a more in-depth morphological and molecular investigation of this genus especially in planktonic habitats.


Assuntos
Diatomáceas/classificação , Diatomáceas/citologia , Proteínas de Algas/análise , Croácia , DNA de Algas/análise , DNA Ribossômico/análise , Diatomáceas/genética , Diatomáceas/ultraestrutura , Mar Mediterrâneo , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Filogenia
5.
Food Technol Biotechnol ; 56(4): 494-505, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30923446

RESUMO

The present study evaluates the potential of encapsulation of polyphenolic antioxidants from rosemary (Rosmarinus officinalis L.) leaves by combining emulsification and spray drying techniques. To stabilize the emulsions and prepare samples suitable for use in dry products, double emulsions encapsulating rosemary polyphenolic extract and containing polyglycerol polyricinoleate (4%), whey protein isolates (2 and 4%) as emulsifiers, and maltodextrins (MDE 10 and 21) as enhancing coatings were subjected to spray drying. The obtained results show insignificant (p>0.05) effect of used maltodextrin type and protein content on mean particle size of double emulsions containing rosemary polyphenols. Morphology analyses showed that double emulsions were successfully prepared, spherical microcapsules were obtained after spray drying of double emulsions and double emulsion form was still preserved after rehydration of spray-dried microcapsules. Regardless of used maltodextrins, significantly (p>0.05) higher encapsulation efficiencies (EE) of total polyphenols (39.57 and 42.83%) in rehydrated samples were achieved when higher protein content (4% whey protein isolate) was used, indicating the major impact of protein content on EE of rosemary polyphenols. Also, using HPLC analysis, rosmarinic and caffeic acids, apigenin and luteolin derivatives were detected among specific polyphenols, where rosmarinic acid had notable encapsulation efficiency ranging from 62.15 to 67.43%. In this way, the obtained microcapsules encapsulating rosemary polyphenols could be easily blended with various dry mixtures, and serve for delivery in different functional products.

7.
Mol Biol Rep ; 44(1): 183-189, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28070706

RESUMO

The aim of the study was to assess forensic pertinence of 12 short tandem repeats (STRs) on X-chromosome in south Croatia population. Investigator® Argus X-12 kit was used to co-amplify 12 STR loci belonging to four linkage groups (LGs) on X-chromosome in 99 male and 98 female DNA samples of unrelated donors. PCR products were analyzed by capillary electrophoresis. Population genetic and forensic parameters were calculated by the Arlequin and POPTREE2 software, and an on-line tool available at ChrX-STR.org. Hardy-Weinberg equilibrium was confirmed for all X-STR markers in female samples. Biallelic patterns at DXS10079 locus were detected in four male samples. Polymorphism information content for the most (DXS10135) and the least (DXS8378) informative markers was 0.9212 and 0.6347, respectively. In both male and female samples, combined power of discrimination exceeded 0.999999999. As confirmed by linkage disequilibrium test, significant association of marker pair DXS10074-DXS10079 (P = 0.0004) within LG2 and marker pair DXS10101-DXS10103 (P = 0.0003) within LG3 was found only in male samples. Number of observed haplotypes in our sample pool amounted 3.01, 7.53, 5 and 3.25% of the number of possible haplotypes for LG1, LG2, LG3 and LG4, respectively. According to haplotype diversity value of 0.9981, LG1 was the most informative. In comparison of south Croatia with 26 world populations, pair-wise [Formula: see text] values increase in parallel with geographical distance. Overall statistical assessment confirmed suitability of Investigator® Argus X-12 kit for forensic casework in both identification and familial testing in the population of south Croatia.


Assuntos
Cromossomos Humanos X/genética , Grupo com Ancestrais do Continente Europeu/genética , Genética Populacional/métodos , Repetições de Microssatélites , Croácia , Feminino , Frequência do Gene , Haplótipos , Humanos , Desequilíbrio de Ligação , Masculino
8.
Genet Mol Biol ; 40(1): 80-83, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-27768154

RESUMO

X chromosome STR typing has emerged recently as a powerful tool, complementary to autosomal STR typing, in solving complex forensic and missing person cases. Investigator® Argus X-12 is a commercial product that allows co-amplification of 12 X chromosomal markers belonging to four linkage groups (LGs). In this study, we analyzed by capillary electrophoresis blood samples from 100 females and 102 males from a population of northern Croatia. Statistical analysis included calculation of allele and haplotype frequencies, as well as forensic parameters. The most informative marker for the northern Croatia population was DXS10135 with PIC=0.9211 and a total of 27 alleles. The least polymorphic marker was DXS8378 with 6 alleles. The proportion of observed haplotypes from the number of possible haplotypes varied from 2.74-8.57% across all LGs, with LG1 being the most informative. Of the 11 tested world populations compared to the population of northern Croatia, significant differences in genetic distance (FST) were found for Greenlandic and all non-European populations. We found that all tested markers are in HWE and can thus be used for match probability calculation. Because of high combined power of discrimination in both men and women, Investigator® Argus X-12 is applicable for the northern Croatia population in routine forensic casework.

9.
Leg Med (Tokyo) ; 21: 77-84, 2016 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-27497338

RESUMO

Investigator® Argus X-12 Kit is a commercially available set that allows simultaneous PCR amplification of 12 X-STR markers belonging to four linkage groups (LG). To assess the forensic efficiency of these markers for the population of central Croatia and consequent applicability in routine forensic casework, DNA from 200 blood samples of unrelated donors (100 female and 100 male) was amplified by Investigator® Argus X-12 Kit and analyzed by capillary electrophoresis. Statistical computations based on allele and haplotype frequencies for LG1 - LG4 were performed using Arlequin 3.5 software and on-line tool available at ChrX-STR.org. In female samples, all X-STR markers were in Hardy-Weinberg equilibrium (HWE). The most informative marker for central Croatia population was DXS10135 with polymorphism information content (PIC) 0.9296. The least polymorphic locus was DXS8378 (PIC=0.6363). Power of discrimination (PD) varied from 0.6968 to 0.9336 in male and from 0.8476 to 0.9916 in female samples. Combined PD exceeded 0.999999999 in both men and women. In male samples, linkage disequilibrium (LD) test revealed significant association (P=0.0000) of one marker pair in LG4 and two marker pairs in LG3. Portion of observed haplotypes in the number of possible haplotypes varied from 2.86% to 7.47% across all LGs. LG1 was the most informative with haplotype diversity (H) 0.9972. High PD of all analyzed markers exhibited for central Croatia population confirms suitability of Investigator® Argus X-12 for forensic pertinence. Moreover, results of this study will be included in establishing a national reference X-STR database based on 12 X-STR loci, which is necessary for the correct interpretation of the forensic casework results.


Assuntos
Cromossomos Humanos X/genética , Impressões Digitais de DNA/métodos , Ligação Genética/genética , Genética Populacional/métodos , Croácia , Impressões Digitais de DNA/instrumentação , Feminino , Técnicas de Genotipagem , Humanos , Masculino
10.
J Forensic Leg Med ; 38: 101-5, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26748240

RESUMO

Various commercially available one-step immunoassays for detection of human (primate) blood have been developed. This study evaluated two hemoglobin tests, ABAcard(®) HemaTrace(®) and HemDirect Hemoglobin against glycophorin A test-RSID™-Blood for following parameters: sensitivity, specificity, effectiveness using various substrates, stain remover and aged blood stains. The highest blood detection limit was observed if HemaTrace(®) was used. When compared with HemaTrace(®), ten times lower sensitivity was observed for HemDirect Hemoglobin test. No false positives were obtained for HemDirect Hemoglobin while ABAcard(®) HemaTrace(®), probably due to its extreme sensitivity, showed high percent of false positives with saliva. The lowest sensitivity and 40% of false positives with saliva was exhibited by RSID™-Blood. In addition, this test encountered the lowest efficacy if aged blood-stains or blood treated with stain remover were used. As expected, none of the tested substrates (wood, metal, brick, and soil), influenced on blood testing, although soil substrate affected STR amplification. Conducted studies established HemDirect Hemoglobin test as more reliable for evaluated parameters than ABAcard(®) HemaTrace(®) and RSID™-Blood.


Assuntos
Manchas de Sangue , Medicina Legal/instrumentação , Impressões Digitais de DNA , Hemoglobinas/isolamento & purificação , Humanos , Repetições de Microssatélites , Reação em Cadeia da Polimerase , Valor Preditivo dos Testes , Sensibilidade e Especificidade
11.
Acta Clin Croat ; 55(2): 279-89, 2016 06.
Artigo em Inglês | MEDLINE | ID: mdl-28394544

RESUMO

The purpose of this study was to create a fibrin-based human skin substitute in vitro with epidermal and dermal component and to assess its healing potential in deep partial and full thickness burns. Fibrin scaffolds were prepared from commercial fibrin glue kits. Human fibroblasts were cultured in fibrin gel. Human keratinocytes were seeded on the top of the gel. Viability of cells was determined fluorimetrically. Scanning electron microscope and immunocytochemistry analysis of cultured cells were performed. After hydrosurgical preparation of deep burn necrotic tissue, wound bed was prepared for skin substitutes. Progress of healing was documented using visual estimation and photos. Scanning electron microscope images showed good cell attachment and colony spreading of keratinocytes and fibroblasts on fibrin scaff old. Immunofluorescent staining of cell cultures on fibrin scaffold showed expression of vimentin, a marker of fibroblast cells, cytokeratin 19, a marker of epithelial stem cells, as well as involucrin, a marker of differentiated keratinocytes. Clinical results clearly showed that appearance of the skin did not differ significantly from the areas of transplanted skin using split-thickness skin graft techniques. In conclusion, using these fibrin-cultured autografts on massive full-thickness burn resulted in good healing.


Assuntos
Queimaduras/terapia , Fibrina , Pele Artificial , Tecidos Suporte , Adolescente , Queimaduras/patologia , Técnicas de Cultura de Células , Sobrevivência Celular , Pré-Escolar , Fibroblastos , Humanos , Queratinócitos , Masculino , Transplante de Pele , Cicatrização
12.
Croat Med J ; 56(3): 246-56, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26088849

RESUMO

AIM: To investigate the impact of synthetic electrospun polyurethane (PU) and polycaprolactone (PCL) nanoscaffolds, before and after hydrolytic surface modification, on viability and differentiation of cultured human eye epithelial cells, in comparison with natural scaffolds: fibrin and human amniotic membrane. METHODS: Human placenta was taken at elective cesarean delivery. Fibrin scaffolds were prepared from commercial fibrin glue kits. Nanoscaffolds were fabricated by electrospinning. Limbal cells were isolated from surpluses of human cadaveric cornea and seeded on feeder 3T3 cells. The scaffolds used for viability testing and immunofluorescence analysis were amniotic membrane, fibrin, PU, and PCL nanoscaffolds, with or without prior NaOH treatment. RESULTS: Scanning electron microscope photographs of all tested scaffolds showed good colony spreading of seeded limbal cells. There was a significant difference in viability performance between cells with highest viability cultured on tissue culture plastic and cells cultured on all other scaffolds. On the other hand, electrospun PU, PCL, and electrospun PCL treated with NaOH had more than 80% of limbal cells positive for stem cell marker p63 compared to only 27%of p63 positive cells on fibrin. CONCLUSION: Natural scaffolds, fibrin and amniotic membrane, showed better cell viability than electrospun scaffolds. On the contrary, high percentages of p63 positive cells obtained on these scaffolds still makes them good candidates for efficient delivery systems for therapeutic purposes.


Assuntos
Córnea , Sistemas de Liberação de Medicamentos/instrumentação , Nanoestruturas/química , Células-Tronco/citologia , Tecidos Suporte , Âmnio/citologia , Animais , Diferenciação Celular , Sobrevivência Celular , Fibrina/química , Humanos , Camundongos , Microscopia Eletrônica de Varredura , Poliésteres/química , Poliuretanos/química
13.
Food Chem ; 167: 378-86, 2015 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-25149001

RESUMO

Alginate-based blends consisting of carrageenan, pectin, chitosan or psyllium husk powder were prepared for assessment of the best formulation aimed at encapsulation of caffeine. Alginate-pectin blend exhibited the lowest viscosity and provided the smallest beads. Alginate-psyllium husk blend was characterised with higher viscosity, yielding the largest bead size and the highest caffeine encapsulation efficiency (83.6%). The release kinetics of caffeine indicated that the porosity of alginate hydrogel was not reduced sufficiently to retard the diffusion of caffeine from the beads. Chitosan coated alginate beads provided the most retarded release of caffeine in water. Morphological characteristics of beads encapsulating caffeine were adversely affected by freeze drying. Bitterness intensity of caffeine-containing beads in water was the lowest for alginate-psyllium beads and chitosan coated alginate beads. Higher sodium alginate concentration (3%) for production of hydrogel beads in combination with psyllium or chitosan coating would present the most favourable carrier systems for immobilization of caffeine.


Assuntos
Alginatos/química , Cafeína/química , Carragenina/química , Quitosana/química , Hidrogel de Polietilenoglicol-Dimetacrilato/análise , Pectinas/química , Psyllium/química , Cafeína/administração & dosagem , Química Farmacêutica , Ácido Glucurônico/química , Ácidos Hexurônicos/química
14.
J Sci Food Agric ; 95(13): 2607-18, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25377798

RESUMO

BACKGROUND: Consumer demands for healthy, functional foods are growing rapidly nowadays. Coffee, as one of the most widespread commodities, represents an interesting aspect for enrichment, since it is consumed by millions of people on a daily basis. The aim of this study was to formulate enriched instant coffee powders with the purpose of estimating the influence of storage time, functional ingredients and packaging material on physical and sensory properties of the mixtures. RESULTS: Storage time of 6 months significantly (P <0.05) influenced moisture content of the mixtures, which rose linearly with an increase in storage time. Packaging material proved to be an important variable affecting moisture content, particle size, colour and cohesion index. Functional ingredients (vitamins A and C, iron, inulin and oligofructose) influenced particle size, dispersibility, wettability and, in terms of sensory analysis, grades for aftertaste, chemical taste and overall acceptability. CONCLUSION: Addition of functional ingredients significantly influenced some particle size distribution parameters and reconstitution properties, causing an increase in wettability and dispersibility times. Furthermore, in sensory terms, it influenced aftertaste and chemical taste grades. Packaging material significantly influenced moisture content, some particle size distribution parameters, colour and cohesion index.


Assuntos
Café , Manipulação de Alimentos/métodos , Alimentos Fortificados , Inulina , Micronutrientes , Oligossacarídeos , Paladar , Bebidas/análise , Cor , Embalagem de Alimentos , Armazenamento de Alimentos , Alimento Funcional , Humanos , Tamanho da Partícula , Pós , Água/análise
15.
Acta Vet Scand ; 55: 54, 2013 Jul 18.
Artigo em Inglês | MEDLINE | ID: mdl-23866945

RESUMO

BACKGROUND: Because European-wide directives are restricting the non-clinical use of antibiotics as in-feed growth promotors in swine production, there is an intensive search for alternative strategies for control and prevention of losses among young pigs. With the growing knowledge of the porcine immune system and its endogenous modulation, it has been clearly established that exogenous immunomodulation using adjuvants and immune response modifiers (IRMs) represents an important prophylactic/therapeutic approach in the prevention/treatment of both stress- and microbial-induced disorders that accompaning weaning. However, it is essential to select a fully evaluated agent which may act either as a nonspecific IRM or synergistically as an adjuvant with vaccines. The synthetic macromolecules with a long history as adjuvant and IRM are nonionic block copolymers which consist of polyoxyethylene (POE) and polyoxypropylene (POP) molecules. METHODS: The aim of this work was to evaluate the effectiveness of POE-POP given as a single peroral dose on productivity parameters such as body weight gain, feed intake and feed conversion ratio, and systemic and intestinal immune parameters by assessing the proportions of CD45+ lymphoid cells, CD4+ and CD8+ T cells, and CD21+ B cells in the peripheral blood as well as the number of CD45RA+ naive lymphoid cells residing in the ileal mucosa in weaned pigs during a follow-up study 5 weeks after the treatment. RESULTS: Pigs treated with POE-POP had better feed intake (+ 14.57%), higher average body mass at the end of the experiment (20.91 kg vs. 17.61 kg), and higher body weight gain in relation to Day 0 (191.63% vs. 144.58%) as well as in relation to nontreated pigs (+ 18.74%), with a lower feed conversion ratio (- 30.26%) in comparison to the control pigs. A much lower diarrhea severity score (5 vs. 54) was recorded in pigs treated with POE-POP (- 90.74%) than in the control pigs. A higher average diarrhea severity (ADS) was recorded in the control pigs (1.54 vs. 0.14), whereas the treatmant group had much a lower ADS ratio (- 90.91%) after 35 days of the experiment. The pigs that were treated with POE-POP had an increased proportion of CD45+, CD4+ and CD8+ cells at Day 21 (at p < 0.05, p < 0.05 or p < 0.01, respectively), Day 28 (at p < 0.01, respectively) and Day 35 (at p < 0.01, p < 0.05 or p < 0.01, respectively) as well as of CD21+ cells at Day 28 (p < 0.05) and Day 35 of the experiment (p < 0.01). Also, these pigs had more numerous CD45RA+ cells in interfollicular (p < 0.05) and follicular areas (p < 0.01) of the ileal Peyer's patches than did control pigs. CONCLUSION: This property of POE-POP to induce recruitment of circulating and intestinal immune cell subsets in weaned pigs may allow the use of IRM-active block copolymers as adjuvants for vaccines, particularly those orally delivered and targeted to the gut-associated lymphoid tissues that are well known to promote rather tolerogenic than protective immune responses.


Assuntos
Polietilenoglicóis/química , Polímeros/química , Propilenoglicóis/química , Suínos/imunologia , Subpopulações de Linfócitos T/fisiologia , Animais , Ganho de Peso
16.
Croat Med J ; 54(3): 238-47, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23771754

RESUMO

AIM: To analyze 8 X-linked short tandem repeat (STR) markers in the population of central Croatia and to evaluate their forensic efficiency. METHODS: We carried out a statistical analysis of the data from previously performed genetic analyses, collected during routine forensic work by the Forensic Science Centre ''Ivan Vucetic.'' Mentype® Argus X-8 PCR amplification kit was used for typing the data of 99 unrelated healthy women and 78 men from central Croatia. Haplotype frequencies were calculated only in male samples. Arlequin 3.5 software was used to assess Hardy-Weinberg equilibrium (HWE), linkage disequilibrium (LD), observed and expected heterozygosity. Power of discrimination (PD) for men and women, polymorphism information content (PIC), power of exclusion, and mean exclusion chance for deficiency cases, normal trios, and duos were determined using online database ChrX-STR.org. RESULTS: In female samples, deviations from HWE (P=0.006) for each locus were not found. LD test performed both on female and male samples revealed no significant association between markers (P=0.002). DXS10135 was the most polymorphic locus (PIC=0.931). PD varied from 0.692 to 0.935 in male and from 0.845 to 0.992 in female samples. Combined PD reached 99.999999% in men and 99.9999999999% in women. CONCLUSION: Performed analyses revealed that the studied marker set contained polymorphic markers with high power of discrimination. We can conclude that Mentype® Argus X-8 PCR is suitable for application in the population of central Croatia. Results of this study, together with collected allele and haplotype frequencies, are the first step in establishing a national reference X-STR database based on 8 X-STR loci.


Assuntos
Cromossomos Humanos X/genética , Frequência do Gene , Genética Populacional , Repetições de Microssatélites/genética , Adulto , Croácia , Feminino , Genes Ligados ao Cromossomo X , Marcadores Genéticos , Haplótipos , Humanos , Desequilíbrio de Ligação , Masculino , Reação em Cadeia da Polimerase , Polimorfismo Genético , Adulto Jovem
17.
Croat Med J ; 54(3): 263-71, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23771757

RESUMO

AIM: To examine the influence of ultraviolet C (UVC) radiation on blood, saliva, semen, and naked DNA samples for preventing DNA cross-contamination on working surfaces in laboratories. METHODS: Blood, saliva, semen, and DNA isolated from buccal swab samples were obtained from a single male donor and applied to the laboratory working surfaces. UVC radiation was applied to these diluted and undiluted samples with or without previous decontamination of the working surfaces with 10% sodium hypochlorite and 20% ethanol. Genomic DNA was extracted using Chelex. After quantification, DNA was amplified using the AmpFlSTR® NGM™ PCR Amplification Kit. We tested and statistically analyzed DNA concentration, UVC dose, sample volume, radiation time, the number of correctly detected alleles on genetic loci, and the number of correctly detected alleles in four groups in which 16 loci were divided. RESULTS: When working surfaces were not decontaminated and were treated only with UVC radiation in the laboratory, the genetic profile for naked DNA could not be obtained after 2 minutes of UVC radiation and for saliva after 54 hours. For blood and semen, a partial genetic profile was obtained even after 250 hours of UVC radiation in the laminar. When working surfaces were decontaminated with 10% sodium hypochlorite and 20% ethanol, genetic profile could not be obtained for naked DNA after 2 minutes, for saliva after 4 hours, for blood after 16 hours, and for semen after 8 hours of UVC radiation in the laboratory. CONCLUSION: It is recommended to carefully and thoroughly clean working surfaces with 10% sodium hypochlorite and 20% ethanol followed by minimal 16-hour UVC exposure (dose approximately 4380 mJ/cm2) for complete and successful decontamination.


Assuntos
Sangue/efeitos da radiação , DNA/efeitos da radiação , Saliva/efeitos da radiação , Sêmen/efeitos da radiação , Raios Ultravioleta , Dano ao DNA , Amplificação de Genes , Humanos , Masculino , Reação em Cadeia da Polimerase , Doses de Radiação , Adulto Jovem
18.
Mol Biol Rep ; 40(7): 4349-60, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23649761

RESUMO

Proper detection and subsequent analysis of biological evidence is crucial for crime scene reconstruction. The number of different criminal acts is increasing rapidly. Therefore, forensic geneticists are constantly on the battlefield, trying hard to find solutions how to solve them. One of the essential defensive lines in the fight against the invasion of crime is relying on DNA methylation. In this review, the role of DNA methylation in body fluid identification and other DNA methylation applications are discussed. Among other applications of DNA methylation, age determination of the donor of biological evidence, analysis of the parent-of-origin specific DNA methylation markers at imprinted loci for parentage testing and personal identification, differentiation between monozygotic twins due to their different DNA methylation patterns, artificial DNA detection and analyses of DNA methylation patterns in the promoter regions of circadian clock genes are the most important ones. Nevertheless, there are still a lot of open chapters in DNA methylation research that need to be closed before its final implementation in routine forensic casework.


Assuntos
Metilação de DNA , Genética Forense , Líquidos Corporais/química , Epigenômica , Marcadores Genéticos , Humanos
19.
Arch Gynecol Obstet ; 286(3): 671-82, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22569709

RESUMO

OBJECTIVE: The objective of this study was determination of causative factors of the genital infections and their correlation with various predictor variables. Secondary objectives included: (1) determination of the presence and the type of low molecular weight metabolites in the samples of vaginal secretion formed in vivo, (2) determination of the concentration of 2-phenylethanol formed in vitro for each Candida species, (3) determination of the relationship between fungal/bacterial/viral infections with the metabolites formed in vivo using multivariate analysis. METHODS: One hundred and ninety-seven women in the age range from 18 to 65 years were included in the study. After the completion of questionnaire, all the patients were subjected to Pap test, cervical swabs for the presence of aerobic bacteria, yeasts, Ureaplasma urealyticum, Chlamydia trachomatis, Mycoplasma, and hrHPV DNA. The presence and the concentration of low-molecular weight metabolites in vitro and in vivo were determined by gas chromatography-mass spectrometry (GC-MS) method. Multivariate analysis methods were used for statistical evaluation. RESULTS: The most important risk factors of fungal/bacterial/viral infections were determined. The presence of 2-phenylethanol in vivo was confirmed in 14 of 74 tested samples and connected with the Candida species. The presence of symptoms, hrHPV DNA and Ureaplasma urealyticum are the predictor variables with the highest influence on the formation of the metabolite in vivo. The results in vitro confirmed that various Candida species produced 2-phenylethanol with the concentrations ranging from 0.6 to 4.64 µg/mL. CONCLUSION: The medical exposure to irradiation, marital status, and number of partners as well as stress factors (miscarriages, chronic, viral, or tumor illnesses) had the highest influence on the development of the bacterial/fungal/viral infections. The formation of 2-phenylethanol, both in vivo and in vitro, was confirmed and connected with Candida species. Besides, according to statistical tests, it seems that presence of symptoms, hrHPV DNA, and Ureaplasma urealyticum had also significant role on the formation of 2-phenylethanol in vivo.


Assuntos
Candida/metabolismo , Álcool Feniletílico/análise , Vagina/microbiologia , Vaginite/microbiologia , Adolescente , Adulto , Idoso , Estudos de Casos e Controles , Croácia/epidemiologia , Feminino , Nível de Saúde , Humanos , Estilo de Vida , Pessoa de Meia-Idade , Esfregaço Vaginal , Vaginite/epidemiologia , Adulto Jovem
20.
Mol Biol Rep ; 39(7): 7727-41, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22391654

RESUMO

A reference Y-chromosome short tandem repeat (STR) haplotype database is needed for Y-STR match interpretation as well as for national and regional characterization of populations. The aim of this study was to create a comprehensive Y-STR haplotype database of the Croatian contemporary population and to analyze substructure between the five Croatian regions. We carried out a statistical analysis of the data from previously performed genetic analyses collected during routine forensic work by the Forensic Science Centre "Ivan Vucetic". A total of 1,100 unrelated men from eastern, western, northern, southern and central Croatia were selected for the purpose of this study. Y-STRs were typed using the AmpFISTR Yfiler PCR amplification kit. Analysis of molecular variance calculated with the Y chromosome haplotype reference database online analysis tool included 16 population samples with 20,247 haplotypes. A total of 947 haplotypes were recorded, 848 of which were unique (89.5%). Haplotype diversity was 0.998, with the most frequent haplotype found in 9 of 1,100 men (0.82%). Locus diversity varied from 0.266 for DYS392 to 0.868 for DYS385. Discrimination capacity was 86.1%. Our results suggested high level of similarity among regional subpopulations within Croatia, except for mildly different southern Croatia. Relative resemblance was found with Bosnia and Herzegovina and Serbia. Whit Atheys' Haplogroup Predictor was used to estimate the frequencies of Y-chromosome haplogroups. I2a, R1a, E1b1b and R1b haplogroups were most frequent in all Croatian regions. These results are important in forensics and contribute to the population genetics and genetic background of the contemporary Croatian population.


Assuntos
Cromossomos Humanos Y/genética , Bases de Dados de Ácidos Nucleicos , Haplótipos , Repetições de Microssatélites , Mapeamento Cromossômico , Croácia , Frequência do Gene , Variação Genética , Genética Populacional , Humanos
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