Usuariu alderique:Oriciu/Archivos/2019

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Agreement of molecular biology and morphology methods in sex determination of human piloter un ulm suisse anti aging from Žatec cemetery 11 th — 13 th century AD Shoda v určení pohlaví při aplikaci molekulárně biologických a morfologických metod na lidské kosterní pozůstatky ze Žatce Sequence length difference in amelogenin gene locus piloter un ulm suisse anti aging both X and Y chromosomes was chosen for its value in sexing.

A modification of the silica method for extraction of aDNA was used. In Obtained data were then confronted with the results given by morphological sexing methods and the concordance of both techniques was This observation can be also taken as an indirect proof of authenticity of extracted aDNA fragments. Extrakce DNA byla provedena pro účely laboratoře upravenou metodou silika. Toto zjištění lze rovněž považovat za nepřímý doklad autenticity DNA izolované ze středověkých kostí.

One of the contributions is sex determination of skeletons, which helps in reconstruction of social structure of past societies. Especially in cases of fragmentary bone remains molecular biology method is the only possibility for informative data retrieving. However the main problem piloter un ulm suisse anti aging aDNA study is contamination of ancient biological samples with modern human DNA that can not be distinguished from the authentic templates Kaestle et al.

Information about sex is hidden in DNA of biological tissues. After long time spent in burial environment aDNA is in very low quantities, fragmented into sequences mainly not longer than bp Cooper et al.

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There are many methods for DNA isolation from hard bone tissue. The phenol-chloroform method Sambrook et al.

It gives similar yields and the extract seems to be free of inhibitory contaminants.

Combination of phenol-chloroform and silica method Krings et al. Therefore, in this study we have optimized original silica isolation method with the use of guanidine thiocyanate as a binding agent Boom ; Höss — Pääbo ; Evison et al. However, in case of female sex there was no positive signal, piloter un ulm suisse anti aging could also mean, that aDNA is not preserved or that the amplification failed.

Another possibility of sex determination is an analysis of amelogenin gene locus present on both X and Y-chromosomes. It has started to be widely used for its positive electrophoresis signals for both sexes Nakahori piloter un ulm suisse anti aging al. Gene for amelogenin protein is located on the pseudoautosomal region Xp There are 5 deletions on X-chromosome 22—80bp and 5 deletions on the Y-chromosome 1—bp Haas-Rochholz — Weiler For aDNA analysis generally the aim is to amplify fragments at lengths up to bp, because aDNA is degraded and fragmented into pieces mainly shorter than bp Pääbo The aim of this study was to establish specific protocol for aDNA isolation that would reveal true uncontaminated results.

For this reason we have taken only adult individuals with sex defined by bone morphology and compared our results to them.

When molecular genetics brings similar results, one can consider such a study as trustworthy. Materials and methods Contamination precautions.

Standard precautions were taken against contamination with intrusive DNA Poinar This room was apart from laboratory, where PCR and electrophoresis took place. All the laboratory equipment was maintained to keep uncontaminated. Plastic gloves were changed frequently. Plastic laboratory ware and glass vessels were autoclaved before use.

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System of blind controls was applied. Sample preparation. We took 19 adult individuals from medieval collection of human remains excavated in Žatec at Chelčického square in years — Those human individuals were chosen for their high preservation of skeletal remains.

Usuariu alderique:Oriciu/Archivos/2019

Higher piloter un ulm suisse anti aging of morphological traits for sex assessment was set as the main criterion in our study where the highest possible accuracy had to be reached. As specimens for genetic analysis fragments of ribs or teeth were taken. We selected tissue samples with good macroscopic status. Teeth had no crevice or visible holes; no openings into spongiosa part were apparent on the rib bone fragments according to Richards et al.

Only compacta was taken as a sample.

2003_4 - Archeologický ústav AV ČR

Specimens were brought to a fine powder by a vibrating ball mill. Extraction of ancient DNA. About 0. Then proteinase K was added and the whole content was kept one more day in motion. DNA was eluted in water and stored in —20°C. Usually the second round of extraction increased aDNA yield to detectable extent. Amplification of nuclear aDNA. Final elongation 72°C was kept for 5 min. We have experienced how difficult it might be to establish optimized PCR protocol for detection of low amounts of degraded aDNA.

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Analyzing of amplified products. Sex determination by morphological approach. Murail et al. The primary diagnosis was undertaken on the group of individuals with preserved pelvic bone — the most reliable part of the skeleton when dealing with a population sample of an unknown extent of sexual dimorphism of extrapelvic dimensions. For primary sex determination we piloter un ulm suisse anti aging chosen both metric Houët et al.

The visual determination of sex from morphological characteristics of the skeletal pelvis is based on five traits Bruzekeach one with firmly defined degrees of feminisation and masculinisation.

The secondary sex diagnosis consisted of calculation of posterior probabilities of discrimination functions statistic package Statistica © based on 31 non-pelvic post-cranial dimensions of the individuals whose sex had been determined by the primary diagnosis. This approach considered the piloter un ulm suisse anti aging of sexual dimorphism of the population under study. Nine dimensions of femur, nine dimensions of tibia, one dimension of fibula, seven dimensions of humerus, one dimension of radius, one dimension of ulna, two dimensions of scapula, and one dimension of clavicle Bräuer figured in the analysis.

Like for primary diagnosis on the snižování velikosti zbavit zaměstnanců, we measured both sides.

For the calculation left side was taken, in case of its absence the observation from right side was used.

Výroční zpráva o činnosti

Results The results of primary and secondary morphological as well as molecular genetic analysis are presented in Table 1. In 18 out of 19 ancient bone samples the extractions were successful. Primary extract supernatant with EDTA and proteinase Kafter the first jednoduchá maska​​ na obličej vrásek had brown colour when it originated from bone specimen and white colour when it had tooth origin.

Disagreement happened in All the three samples were only once determined. Two recept na zvlhčování proti stárnutí of these three samples were genetically determined piloter un ulm suisse anti aging women with visible band of 80bp.

Where two or more samples of one individual were proceeded simultaneously, there was a concordance among the amounts of amplified products. Samples of bone and tooth of one individual were never subjected to the same aDNA isolation process.

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Discussion We have successfully developed an aDNA extraction protocol for retrieving information about sex of individuals from 11 th — 13 th century. In 18 out of 19 This ratio is higher than average but still falls into range previously obtained by other aDNA studies.

In a study where genetic and morphological methods were also compared aDNA was extracted in 12 out of 42 In a study of Vernesi et al. Inconsistencies between morphometric and genetic sex data were also found in a study, where 3 out of 5 buried individuals revealed different sex when both methods were applied Götherström et al.

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The confrontation of morphological and molecular genetic results of sex determination of 19 individuals from Žatec necropolis. AO — inventory number of the skeleton; pF — probability of female sex by primary diagnosis metric approach ; pM — probability of male sex by primary piloter un ulm suisse anti aging metric approach ; visual — result of visual approach of primary diagnosis; SD1 f m — female male posterior probability of discriminant function based on the anterio-posterior diameter of femur diaphysis combined with the maximal length of tibia; SD2 f m — female male posterior probability of discriminant function based on the diameter of femoral head combined with the maximal diameter in the middle of humerus; SD3 f m — female male posterior probability of discriminant function based on the maximal length of radius combined with the maximal length of the articular facet of scapula; GENE — molecular genetic testing by amelogenin locus; nd — data not available.

Konfrontace morfologických a molekulárně genetických výsledků určení pohlaví 19 jedinců z žateckého pohřebiště. AO — inventární číslo kostry; pF — pravděpodobnost ženského pohlaví primární diagnózou česnek proti stárnutí přístup ; pM — pravděpodobnost mužského pohlaví primární diagnózou metrický přístup ; visual — výsledek morfoskopické metodiky primární diagnózy; SD1 f m — posteriorní pravděpodobnost ženského mužského pohlaví u diskriminační funkce založené na předozadním průměru diafýzy kosti stehenní kombinované s maximální délkou kosti holenní; SD2 f m — posteriorní pravděpodobnost ženského mužského pohlaví u diskriminační funkce založené na průměru hlavice kosti stehenní kombinované s maximálním průměrem středu kosti ramenní; SD3 f m — posteriorní pravděpodobnost ženského mužského pohlaví u diskriminační funkce založené na maximální délce kosti vřetení kombinované s maximální délkou artikulační plochy lopatky; GENE — molekulárně genetické testování amelogeninového lokusu; nd — údaj není piloter un ulm suisse anti aging dispozici.

In our study, the results of sex determination correlated in The system of sex assignment might reach only two values — male or female. The chance that false positive results happened in our study was calculated to 0.

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In a case where inconsistencies are found it is hard to determine which method gave the wrong result. All the morphological sex determination techniques have been based on the study of contemporary populations.

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Nivea Zmatňující denní krem a correct application, the characters analyzed should be calibrated to the population examined Vernesi et al. Higher degree of agreement of morphological and genetic results observed in our study can be explained both by the selection of individuals with clear-cut sex traits and by the application of more sophistical statistical approaches of morphological assessments. The concept of primary and secondary diagnosis has been tested on the samples of different biological origins where reliable results have been achieved.

piloter un ulm suisse anti aging

The major advantage of this approach is that it takes into consideration specific sexual dimorphism of the population under study. It should be noted that this concept has not been followed in other above-mentioned studies.

Transkript 1 Výroční zpráva o činnosti 2 předkládá: doc. Boris Kreuzberg, CSc. Zároveň je část lékařů, pracujících na Lékařské fakultě UK v Plzni, vytížena léčebně preventivní činností na půdě Fakultní nemocnice v Plzni, kde většinou zastávají klíčové pozice na jednotlivých klinikách. Pedagogická činnost probíhá ve dvou základních programech Všeobecné lékařství a Zubní lékařství v českém a anglickém jazyce.

The protocol developed for our study seems to be very useful tool for sex analysis. It is more likely to obtain shorter fragments among degraded aDNA sequences. Our extracts were free of exogenous contaminating DNA as we can deduce from the system of blind controls and high percentage of sex agreement between morphological Zbavte se rychle Zits genetic data.

According to some findings, blank controls without amplification product do not mean that contamination was absent in other tubes Kolman So the disagreement of both methods could be also due to hidden contamination.

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It could also happen due to the fact that the sample under study belonged mistakenly to someone else. We cannot exclude that the burial findings of more individuals in one grave could not be mixed together. Burials on the medieval cemeteries are placed in layers often not well separated from each other. Female identification in a case where morphologic method defined man occurred twice in our study. This phenomenon might be explained by all above mentioned and also by allelic drop-out where one of alleles is missing in the extracted sample Zierdt et al.

We have experienced difficulties with aDNA extraction. It has shown to us that the second round of extraction from the bone powder previously used brought better results. It could be due to longer storage of sample with residual EDTA combined with proteinase K in some cases about one year at 4°C. The reason for better result could also be due to the fact, that in the first extraction from bone powder all the contaminating chemicals such as proteins or humic acids Hagelberg — Clegg are reduced to minimum level allowing the extract to be cleaner and better amplifiable.

We cannot confirm that aDNA from tooth tissue is preserved in higher quality status than in bone material Kurosaki et al. Samples of bone and tooth of one individual were never subjected to the same aDNA isolation process so it is hard to determine if tooth material possess higher or cleaner status of aDNA. Higher amounts of amplified products from aDNA that originated from teeth were noticeable though.

This study highlights the protocol for aDNA isolation from bone and teeth archaeological material.