Henryk W. Witas

The anti-asparagines antibodies correlate with l-asparagines activity and may affect clinical outcome of childhood acute lymphoblastic leukemia

The primary aim of the study was to evaluate the importance of anti-asparaginase antibodies for l-asparaginase activity in children with standard and medium risk acute lymphoblastic leukemia (ALL). Forty-seven children with newly diagnosed ALL were included into the prospective study. Enzyme activity and the presence of anti-asparaginase antibodies (IgG and IgM class) were determined. Anti-asparaginase antibodies were identified in 13/47 (IgM class) and 10/47 (IgG class) patients in the induction and in 19/47 (IgM class) and 20/47 (IgG class) patients in the reinduction phase of treatment. The enzyme activity was lower in patients that were positive for anti-asparaginase antibodies, especially in reinduction phase (median 37 (20 – 180) vs 355 (141 – 499), p = 0.001). An association between anti-asparaginase antibodies and the allergic reaction to the drug was found. Besides, the children who developed anti-asparaginase antibodies in the induction phase of treatment showed lower event-free survival as well as overall survival in comparison with children without antibodies. Since our study was carried out in a small number of patients, this observation is only speculative and needs to be confirmed by a further study on a larger sample size, with multivariable analysis. However, our data suggest that l-asparaginase activity together with anti-asparaginase antibodies measurements may become useful for effective therapy of ALL.

mtDNA from the Early Bronze Age to the Roman Period Suggests a Genetic Link between the Indian Subcontinent and Mesopotamian Cradle of Civilization

Ancient DNA methodology was applied to analyse sequences extracted from freshly unearthed remains (teeth) of 4 individuals deeply deposited in slightly alkaline soil of the Tell Ashara (ancient Terqa) and Tell Masaikh (ancient Kar-Assurnasirpal) Syrian archaeological sites, both in the middle Euphrates valley. Dated to the period between 2.5 Kyrs BC and 0.5 Kyrs AD the studied individuals carried mtDNA haplotypes corresponding to the M4b1, M49 and/or M61 haplogroups, which are believed to have arisen in the area of the Indian subcontinent during the Upper Paleolithic and are absent in people living today in Syria. However, they are present in people inhabiting today’s Tibet, Himalayas, India and Pakistan. We anticipate that the analysed remains from Mesopotamia belonged to people with genetic affinity to the Indian subcontinent since the distribution of identified ancient haplotypes indicates solid link with populations from the region of South Asia-Tibet (Trans-Himalaya). They may have been descendants of migrants from much earlier times, spreading the clades of the macrohaplogroup M throughout Eurasia and founding regional Mesopotamian groups like that of Terqa or just merchants moving along trade routes passing near or through the region. None of the successfully identified nuclear alleles turned out to be ΔF508 CFTR, LCT-13910T or Δ32 CCR5.

Between the Baltic and Danubian Worlds: The Genetic Affinities of a Middle Neolithic Population from Central Poland

For a long time, anthropological and genetic research on the Neolithic revolution in Europe was mainly concentrated on the mechanism of agricultural dispersal over different parts of the continent. Recently, attention has shifted towards population processes that occurred after the arrival of the first farmers, transforming the genetically very distinctive early Neolithic Linear Pottery Culture (LBK) and Mesolithic forager populations into present-day Central Europeans. The latest studies indicate that significant changes in this respect took place within the post-Linear Pottery cultures of the Early and Middle Neolithic which were a bridge between the allochthonous LBK and the first indigenous Neolithic culture of north-central Europe—the Funnel Beaker culture (TRB). The paper presents data on mtDNA haplotypes of a Middle Neolithic population dated to 4700/4600–4100/4000 BC belonging to the Brześć Kujawski Group of the Lengyel culture (BKG) from the Kuyavia region in north-central Poland. BKG communities constituted the border of the “Danubian World” in this part of Europe for approx. seven centuries, neighboring foragers of the North European Plain and the southern Baltic basin. MtDNA haplogroups were determined in 11 individuals, and four mtDNA macrohaplogroups were found (H, U5, T, and HV0). The overall haplogroup pattern did not deviate from other post-Linear Pottery populations from central Europe, although a complete lack of N1a and the presence of U5a are noteworthy. Of greatest importance is the observed link between the BKG and the TRB horizon, confirmed by an independent analysis of the craniometric variation of Mesolithic and Neolithic populations inhabiting central Europe. Estimated phylogenetic pattern suggests significant contribution of the post-Linear BKG communities to the origin of the subsequent Middle Neolithic cultures, such as the TRB.

Hunting for the LCT-13910*T Allele between the Middle Neolithic and the Middle Ages Suggests Its Absence in Dairying LBK People Entering the Kuyavia Region in the 8th Millennium BP

Populations from two medieval sites in Central Poland, Stary Brześć Kujawski-4 (SBK-4) and Gruczno, represented high level of lactase persistence (LP) as followed by the LCT-13910*T allele’s presence (0.86 and 0.82, respectively). It was twice as high as in contemporaneous Cedynia (0.4) and Śródka (0.43), both located outside the region, higher than in modern inhabitants of Poland (0.51) and almost as high as in modern Swedish population (0.9). In an attempt to explain the observed differences its frequency changes in time were followed between the Middle Neolithic and the Late Middle Ages in successive dairying populations on a relatively small area (radius ∼60km) containing the two sites. The introduction of the T allele to Kuyavia 7.4 Ka BP by dairying LBK people is not likely, as suggested by the obtained data. It has not been found in any of Neolithic samples dated between 6.3 and 4.5 Ka BP. The identified frequency profile indicates that both the introduction and the beginning of selection could have taken place approx. 4 millennia after first LBK people arrived in the region, shifting the value of LP frequency from 0 to more than 0.8 during less than 130 generations. We hypothesize that the selection process of the T allele was rather rapid, starting just after its introduction into already milking populations and operated via high rates of fertility and mortality on children after weaning through life-threatening conditions, favoring lactose-tolerant individuals. Facing the lack of the T allele in people living on two great European Neolithization routes, the Danubian and Mediterranean ones, and based on its high frequency in northern Iberia, its presence in Scandinavia and estimated occurrence in Central Poland, we propose an alternative Northern Route of its spreading as very likely. None of the successfully identified nuclear alleles turned out to be deltaF508 CFTR.

Frequency of lactase persistence genotype in a healthy Polish population

The majority of mammals are unable to digest lactose due to post-weaning deactivation of the LCT gene, which is responsible for encoding the enzyme lactase (i.e., adult-type hypolactasia). A substitution of C with T at position −13910 bp upstream of the LCT gene has been linked to the lactase persistence phenotype in European populations. We investigated the frequency of LCT-13910C>T polymorphism in 223 blood donors from central Poland. All samples were genotyped by polymerase chain reaction and direct sequencing. The LCT-13910 T allele (lactase persistence) was present in 51% of individuals sampled from the Polish population. We did not find any non-European variants associated with lactase persistence (LCT-13907C>G, LCT-13913T>C, LCT-13915T>G), or any new polymorphisms within the sequenced region. Allele frequencies obtained are in agreement with results from other countries and confirm the unique pattern of distribution of the LCT-13910C>T genotype in Europe.

Do determinants of platelet function co-segregate with genetic markers of type 1 diabetes mellitus?: Analysis of platelet and fibrinolytic parameters in families with type 1 diabetes mellitus

This study examined the significance of selected parameters of primary haemostasis to discriminate between relatives of children with insulin-dependent diabetes mellitus (IDDM). Platelet function, including markers of spontaneous and agonist-induced platelet activation (CD62), platelet consumption (microparticles) and clumping (aggregates), as well as selected parameters of the fibrinolytic system (t-PA and PAI-1), were studied in IDDM children ( n = 45), their parents ( n = 65), siblings ( n = 17) and unrelated healthy controls ( n = 51). The fraction of activated platelets circulating in whole blood amounted to 4.3 +/- 2.1% in IDDM children, and significantly exceeded the level found in parents (1.3 +/- 0.7%, P < 0.002), siblings (1.2 +/- 1.0%, P < 0.002), and controls (1.2 +/- 0.6%, P < 0.002). Furthermore, an enhanced formation of platelet microparticles was observed in the IDDM group, both in resting platelets and also when platelets were stimulated with thrombin. Significantly decreased total PAI-1 occurred in IDDM children ( P < 0.02 versus parents); also slightly lowered active PAI-1 and t-PA antigen were noticed in IDDM subjects compared to other groups, however, the differences were not statistically significant. To assess dissimilarities between the groups of subjects we applied the forward stepwise model of discriminant function analysis, which included platelet flow cytometry parameters. The best separation and the highest discrepancy (expressed as the so called squared Mahalanobis distances, d ) was M revealed between controls and IDDM patients ( P < < 0.0001) and between controls and parents ( P < < 0.0001). The values of d found between IDDM children and their siblings (P < 0.001), as well as parents ( P < 0.01), were M of much lower significance. The finding that the control group, representing unrelated subjects, remains particularly well separated from the other groups, more or less clustered together, implies the possible involvement of genetic factor(s) which might potentially affect platelet activation and reactivity. In addition, the distinguished distribution of HLA DQAI(52) and HLA DQBI(57) genotypes in the groups further validates the suspicion that the altered platelet function and response in diabetes might be associated with some independent genetic factor(s), and is not likely to result from HLA DQAI(52) and HLA DQBI(57) impact.

The Late Neolithic sepulchral and ritual place of site 14 in Kowal (Kuyavia, Central Poland)

Zusammenfassung: Forschungsgegenstand dieses Artikels ist ein Begräbnis- und Ritualplatz der Kugelamphoren-Kultur der Fundstätte 14 in Kowal (Zentral-Polen). Die Stätte umfasst einen Submegalithen sowie einen Ritualplatz mit Tiergräbern und einer menschlichen Bestattung. Der Komplex kann auf die Zeit zwischen 3250/3100 und 2400/2150 v. Chr. datiert werden. Analysen haben gezeigt, dass der Platz über eine lange Zeit benutzt wurde, und dass die durchgeführten Rituale sehr komplex waren. Beweise wurden gefunden, dass an einigen Stellen rituelle Handlungen ohne Unterbrechung über viele Jahre durchgeführt wurden und einzelne Tieropfer und Niederlegungen nur durch einen Zeitraum von wenigen Jahren voneinander getrennt waren. Die Fund-Struktur einiger Gruben zeigt eine sehr penible Organisation des geheiligten Platzes und ermöglicht an einigen Stellen sogar eine Unterscheidung in Nutz- und rituelle Zonen. Weitere wichtige Informationen wurden durch die Untersuchung einer menschlichen Grablege an dieser Stätte gewonnen. Die Analyse der Grabbeigaben hat deren Natur als rituelle Gaben gezeigt; weiter weist sie auf eine dominante Rolle von Tieren im täglichen Leben der verstorbenen Person hin. Wichtige Daten wurden durch die paläo-biologische Untersuchung der menschlichen Knochen gewonnen. Die paläo-serologische Analyse ergab die Blutgruppe der bestatteten Person. Dies ist der erste derartige Nachweis in der Geschichte der europäischen neolithischen Forschung. Bei den Ergebnissen der DNA-Analyse handelt es eines Vertreters der Kugelamphoren-Kultur in der Literatur und zudem um eine von nur wenigen solcher Untersuchungen eines neolithischen Menschen. Einige der nachgewiesenen Allele sind die ältesten Beispiele dieses speziellen Allel-Typs, die in menschlichen Überresten bis heute identifiziert wurden. Die DNA-Analysen belegen eine Laktose-Intoleranz der bestatteten Person, sie legen weiter eine genetische Verwandtschaft mit der Bevölkerung aus der Region des Fruchtbaren Halbmondes nahe. Die Analyse des stabilen Sauerstoff-Isotopen-Verhältnisses zeigt Muster für häufigeren Ortswechsel und das Entwöhnungsalter. Analysen von stabilen Kohlenstoff- und Stickstoff-Isotopen-Verhältnissen haben Rückschlüsse auf die Ernährung ermöglicht; besondere Aufmerksamkeit verdient die Tatsache, dass bis zu 25 % der Kohlenhydrat-haltigen Nahrung aus Hirse bestanden hat. Das ist der früheste Isotopen-gestützte Nachweis von Hirse in der menschlichen Ernährung in Mitteleuropa. Résumé: Le site 14 de Kowal en Pologne centrale, un complexe rituel appartenant à la culture des amphores globulaires, forme le sujet de cet article. Le site comporte un sub-mégalithe, une structure rituelle avec des dépôts d’animaux et une sépulture humaine. Il fut occupé pendant une longue période allant de 3250/3100 à 2400/2150 av. J.-C. et les rituels étaient complexes: certains furent célébrés au même endroit pendant longtemps, tandis que les sacrifices et dépôts d’animaux n’étaient séparés que par de courts intervalles de peutêtre quelques années. Le remplissage de plusieurs fosses démontre une organisation précise de l’espace sacré, et il a même été possible de faire une distinction entre les actes de déposition rituelle et les dépôts de type utilitaire à l’intérieur de ces fosses. L’étude de la sépulture humaine a également fourni d’importants indices. Il en ressort qu’un individu intentionnellement choisi parmi les membres de ‘l’aristocratie’ locale a été la victime d’un meurtre rituel. L’analyse du mobilier funéraire indique qu’il s’agit d’offrandes rituelles et suggère que les animaux jouaient un rôle déterminant dans la vie de cet individu. L’étude de ses ossements a fourni un important ensemble de données, y compris les résultats d’une analyse sérologique permettant d’identifier – pour la première fois pour le Néolithique européen – le groupe sanguin de cette homme. Dans une étude également pionnière pour la culture des amphores globulaires, l’analyse ADN a permis de décrire la composition génétique d’un de ses membres, une rareté dans les études sur le Néolithique. Certains allèles sont les plus anciens exemplaires d’un type particulier d’allèles jusqu’à présent découvert dans des restes humains. Les données de l’analyse de l’ADN de cet individu suggèrent qu’il était intolérant au lactose et qu’il avait des affinités génétiques avec le Croissant fertile. Les résultats de l’analyse des isotopes stables de l’oxygène peuvent être interprétés soit en termes de mobilité ou de sevrage. Ceux provenant de l’analyse des isotopes stables du carbone et de l’azote nous permettent de reconstruire l’alimentation de notre individu ; en particulier le millet fournissait jusqu’à 25 % de sa consommation en glucides. Ce résultat constitue le premier indice isotopique de la présence du millet dans l’alimentation des populations de l’Europe centrale. Abstract: Site 14 in Kowal (central Poland), a funerary and ritual place of the Globular amphora culture forms the subject of this article. The site includes a sub-megalith, a ritual feature with animal burials and a human burial. The complex can be dated to a period spanning from 3250/3100 to 2400/2150 BC; it was used over a long period, and the rituals carried out were complex. Rituals were performed over many years in certain areas, while individual acts of animal sacrifice and deposition may have been separated by a short interval of just a few years. The fill of some pits shows that the sacred space was carefully organised and it has been possible to distinguish between some utilitarian and ritual deposits within them. Other important information is provided by the study of a human burial discovered at the site. It is possible that a deliberately chosen individual from the tribal ‘aristocracy’ was the victim of a ritual murder. The analysis of the grave goods reveals that they were ritual offerings, and these suggest that animals played a dominant role in the deceased man’s daily life. The study of his bones provides important new insights, including those from a palaeo-serological analysis revealing his blood group, a first in the history of European Neolithic research. The results of DNA analysis constitute the first description of the genetic traits of a representative of the Globular amphora culture, among very few such DNA profiles available for a Neolithic person. Some of the alleles discovered are the oldest examples of particular alleles among those identified in human remains so far. DNA evidence suggests that the individual was lactose intolerant, and it demonstrates a genetic relationship with the Fertile Crescent. The analysis of stable oxygen isotope ratios shows patterns compatible with either residential mobility or weaning age. The analysis of stable carbon and nitrogen isotope ratios allowed us to reconstruct the individual’s diet; particular attention is drawn to the fact that up to 25 % of his carbohydrate diet consisted of millet. This is the earliest isotopic evidence for the presence of millet in the diet of people living in Central Europe.

Extremely high frequency of autoimmune-predisposing alleles in medieval specimens

The precise etiology and reasons for the increase in incidence of autoimmune disorders still remain unclear, and although both genetic and environmental factors have been proven to shape individual predisposition, it is not known which of the factors, if not both, is responsible for the boom observed during the last decades. In order to establish whether a higher frequency of autoimmune-predisposing alleles may explain this increase we took advantage of ancient DNA methodology to establish the genetic predisposition, conferred by cytotoxic T lymphocyte associated antigen-4 (CTLA4) +49A/G and human leukocyte antigens (HLA) DQB157, in population inhabiting Poland in the Middle Ages. After successful typing of 42 individuals from a 12th∼14th’s century archeological burial site, we found that frequencies of the predisposing alleles in the medieval population were higher than they are at present, suggesting thus that the recently observed incidence increase results most probably from factors of other than genetic nature.

Origin of the ornamented bâton percé from the Gołębiewo site 47 as a trigger of discussion on long-distance exchange among Early Mesolithic communities of Central Poland and Northern Europe

This article describes evidence for contact and exchange among Mesolithic communities in Poland and Scandinavia, based on the interdisciplinary analysis of an ornamented bâton percé from Gołębiewo site 47 (Central Poland). Typological and chronological-cultural analyses show the artefact to be most likely produced in the North European Plain, during the Boreal period. Carbon-14 dating confirms the antiquity of the artefact. Ancient DNA analysis shows the artefact to be of Rangifer tarandus antler. Following this species designation, a dispersion analysis of Early-Holocene reindeer remains in Europe was conducted, showing this species to exist only in northern Scandinavia and north-western Russia in this period. Therefore, the bâton from Gołębiewo constitutes the youngest reindeer remains in the European Plain and south-western Scandinavia known to date. An attempt was made to determine the biogeographic region from which the antler used to produce the artefact originates from. To this end, comprehensive δ18O, δ13C and δ15N isotope analyses were performed. North Karelia and South Lapland were determined as the most probable regions in terms of isotopic data, results which correspond to the known distribution range of Rangifer tarandus at this time. In light of these finds, the likelihood of contact between Scandinavia and Central Europe in Early Holocene is evaluated. The bâton percé from Gołębiewo is likely key evidence for long-distance exchange during the Boreal period.

Do determinants of platelet function co-segregate with genetic markers of type 1 diabetes mellitus?

: This study examined the significance of selected parameters of primary haemostasis to discriminate between relatives of children with insulin-dependent diabetes mellitus (IDDM). Platelet function, including markers of spontaneous and agonist-induced platelet activation (CD62), platelet consumption (microparticles) and clumping (aggregates), as well as selected parameters of the fibrinolytic system (t-PA and PAI-1), were studied in IDDM children ( n = 45), their parents ( n = 65), siblings ( n = 17) and unrelated healthy controls ( n = 51). The fraction of activated platelets circulating in whole blood amounted to 4.3 +/- 2.1% in IDDM children, and significantly exceeded the level found in parents (1.3 +/- 0.7%, P < 0.002), siblings (1.2 +/- 1.0%, P < 0.002), and controls (1.2 +/- 0.6%, P < 0.002). Furthermore, an enhanced formation of platelet microparticles was observed in the IDDM group, both in resting platelets and also when platelets were stimulated with thrombin. Significantly decreased total PAI-1 occurred in IDDM children ( P < 0.02 versus parents); also slightly lowered active PAI-1 and t-PA antigen were noticed in IDDM subjects compared to other groups, however, the differences were not statistically significant. To assess dissimilarities between the groups of subjects we applied the forward stepwise model of discriminant function analysis, which included platelet flow cytometry parameters. The best separation and the highest discrepancy (expressed as the so called squared Mahalanobis distances, d ) was M revealed between controls and IDDM patients ( P < < 0.0001) and between controls and parents ( P < < 0.0001). The values of d found between IDDM children and their siblings (P < 0.001), as well as parents ( P < 0.01), were M of much lower significance. The finding that the control group, representing unrelated subjects, remains particularly well separated from the other groups, more or less clustered together, implies the possible involvement of genetic factor(s) which might potentially affect platelet activation and reactivity. In addition, the distinguished distribution of HLA DQAI(52) and HLA DQBI(57) genotypes in the groups further validates the suspicion that the altered platelet function and response in diabetes might be associated with some independent genetic factor(s), and is not likely to result from HLA DQAI(52) and HLA DQBI(57) impact.