Donor-derived DNA in hair follicles of recipients after allogeneic hematopoietic stem cell transplantation.

The hair follicles of recipients of allogeneic hematopoietic SCT (HSCT) constitute the tissue with the greatest need for regeneration after high-dose chemotherapy. Previous studies have shown a lack of donor-derived DNA in the hair follicles of recipients. Therefore, we carried out a study to determine whether male donor-derived genetic material can be found in female recipients' hair follicles after HSCT. Fluorescent-based PCR with analyses of Y-chromosome STR (Y-STR) and RQ-PCR with the sex-determining region Y (SRY) were used independently to evaluate chimerism status. Our results proved the existence of donor-derived stem DNA in the recipients' hair follicle cells. This report undermines the validity of data indicating that hair follicle cells maintain 100% of recipient origin.

Association studies of CTLA-4, CD28, and ICOS gene polymorphisms with B-cell chronic lymphocytic leukemia in the Polish population.

Abnormal expression of the costimulatory molecules cytotoxic T-lymphocyte antigen 4 (CTLA-4), CD28, and inducible co-stimulator (ICOS) leads to disturbances of immune response and an increased risk of cancer. An extended study was undertaken to evaluate the association among the polymorphisms CTLA-4c.49A>G, CTLA-4g.319C>T, CTLA-4g.*642AT(8_33), CD28c.17+3T>C, and ICOSc.1554+4GT(8_15) and susceptibility to B-cell chronic lymphocytic leukemia (B-CLL) in the Polish population. The study revealed increased frequency of the CTLA-4g.319C>T [T] allele and the CTLA-4g.319C>T [T] phenotype in B-CLL patients compared with healthy controls (p = 0.003, odds ratio [OR] = 1.73; and p = 0.009, OR = 1.74, respectively). The presence of the CD28c.17+3T>C [C] allele and the CD28c.17+3T>C [C] phenotype increased the OR of B-CLL to 1.59 (p = 0.007) and 1.74 (p = 0.007), respectively. Either CTLA-4g.319C>T or CD28c.17+3T>C was associated with time to Rai stage progression. The distributions of the alleles and genotypes of the ICOS gene significantly differed between patients and controls (p = 0.0009 and p = 0.006, respectively). Individuals possessing short alleles were 2.02 times more prone to B-CLL than others (p = 0.001), whereas carriers of long alleles were protected from B-CLL (p = 0.02, OR = 0.62). The haplotype association study and multivariate analysis confirmed the association of CTLA-4g.319C>T and ICOSc.1554+4GT(8_15) gene polymorphisms with B-CLL. The polymorphic sites CTLA-4c.49A>G and CTLA-4g.*642AT(8_33) did not correlate with B-CLL. Our results are the first in the literature to report that gene polymorphism of the costimulatory molecules CTLA-4, CD28, and ICOS contributes to susceptibility to B-CLL.

Y-SNP-genotyping - a new approach in forensic analysis.

Y-chromosomal DNA polymorphisms, especially Y-STRs are well established in forensic routine case work. The STRs are used for identification in paternity deficiency cases and stain analysis with complicate mixtures of male and female DNA. In contrast, Y-chromosomal SNPs are a new tool in forensic investigations. At present, Y-SNPs are mainly used in molecular anthropology for evolutionary studies. Nevertheless, these markers could also provide very useful information for the analysis of forensic cases. The aim of the presented study was to test Y-SNP-typing for stain analyses using different methods-SNaPshot and MALDI-TOF MS. Both methods are based on the principle of minisequencing. The selected Y-SNP markers are suited to define the most important European haplogroups.

Progression of multiple sclerosis is associated with exon 1 CTLA-4 gene polymorphism.

OBJECTIVES: Multiple sclerosis (MS) is a chronic demyelinating disease of the central nervous system which is widely believed to have a T-cell-mediated etiology. The cytotoxic T-lymphocyte antigen-4 (CTLA-4) antigen molecule plays a key role in the downregulation of T-cell responses. To examine the genetic association of the CTLA-4 gene locus with MS, we analyzed an exon 1 (A49G) transition. MATERIAL AND METHODS: One hundred and fifty-two MS patients and 154 controls were examined. The A/G transition was genotyped by a polymerase chain reaction followed by labeling with a SNaPshot kit and detection using a capillary genetic analyzer. RESULTS: The genotype, allele and phenotype frequencies did not differ significantly between MS patients and controls. Those MS patients with AA and AG genotypes had 4.36 times greater risk of progressing from the relapsing-remitting to the secondary progressive form of the disease than those with the GG genotype. CONCLUSION: The results of our study indicate that CTLA-4 (A49G) exon 1 polymorphism is associated with MS progression.

Online reference database of European Y-chromosomal short tandem repeat (STR) haplotypes.

The reference database of highly informative Y-chromosomal short tandem repeat (STR) haplotypes (YHRD), available online at http://ystr.charite.de, represents the largest collection of male-specific genetic profiles currently available for European populations. By September 2000, YHRD contained 4688 9-locus (so-called "minimal") haplotypes, 40% of which have been extended further to include two additional loci. Establishment of YHRD has been facilitated by the joint efforts of 31 forensic and anthropological institutions. All contributing laboratories have agreed to standardize their Y-STR haplotyping protocols and to participate in a quality assurance exercise prior to the inclusion of any data. In view of its collaborative character, and in order to put YHRD to its intended use, viz. the support of forensic caseworkers in their routine decision-making process, the database has been made publicly available via the Internet in February 2000. Online searches for complete or partial Y-STR haplotypes from evidentiary or non-probative material can be performed on a non-commercial basis, and yield observed haplotype counts as well as extrapolated population frequency estimates. In addition, the YHRD website provides information about the quality control test, genotyping protocols, haplotype formats and informativity, population genetic analysis, literature references, and a list of contact addresses of the contributing laboratories.

Microsatellite instability in thyroid papillary carcinoma and multinodular hyperplasia.

Microsatellite instability (MSI) is a molecular landmark of mutations in DNA mismatch repair genes. The impaired efficiency of DNA repair mechanisms promotes carcinogenesis as well as contributes to tumour progression. Until now, only few studies on MSI in thyroid tumours have been published. Therefore, the aim of the present study was to investigate MSI as a possible characteristic feature of thyroid tumours. The analysis of 12 thyroid papillary carcinomas and 17 multinodular hyperplasias at 13 microsatellite loci showed MSI and loss of heterozygosity (LOH) in both types of lesion, with more alterations noted in the papillary thyroid carcinomas (in 65%) than in multinodular hyperplasia (in 35%). In carcinomas, LOH occurred more frequently than MSI, while in multinodular hyperplasia the LOH/MSI ratio is almost equal.

Characteristics and frequency of germline mutations at microsatellite loci from the human Y chromosome, as revealed by direct observation in father/son pairs.

A number of applications of analysis of human Y-chromosome microsatellite loci to human evolution and forensic science require reliable estimates of the mutation rate and knowledge of the mutational mechanism. We therefore screened a total of 4,999 meioses from father/son pairs with confirmed paternity (probability >/=99. 9%) at 15 Y-chromosomal microsatellite loci and identified 14 mutations. The locus-specific mutation-rate estimates were 0-8. 58x10-3, and the average mutation rate estimates were 3.17x10-3 (95% confidence interval [CI] 1.89-4.94x10-3) across 8 tetranucleotide microsatellites and 2.80x10-3 (95% CI 1.72-4.27x10-3) across all 15 Y-chromosomal microsatellites studied. Our data show a mutational bias toward length increase, on the basis of observation of more repeat gains than losses (10:4). The data are in almost complete agreement with the stepwise-mutation model, with 13 single-repeat changes and 1 double-repeat change. Sequence analysis revealed that all mutations occurred in uninterrupted homogenous arrays of >/=11 repeats. We conclude that mutation rates and characteristics of human Y-chromosomal microsatellites are consistent with those of autosomal microsatellites. This indicates that the general mutational mechanism of microsatellites is independent of recombination.

[Apolipoprotein E(ApoE)and tau protein in Alzheimer type dementia]. / Apolipoproteina (ApoE) i bialko tau w otepieniu typu Alzheimera.

In 45 patients meeting NINCDS-ARDRA criteria for probably diagnosis of Alzheimer disease (AD), ApoE genotype and tau protein level in cerebro-spinal fluid (CSF) were determined. Frequency of e4 allele occurrence in group of AD patients was 73.3% and showed high statistic significance in comparison with control group. Increase of tau protein level in CSF was also statistically significant. No correlation in ApoE allele and tau protein level in CSF was revealed. The authors emphasize the usefulness of tau protein level measurement and determination of ApoE allele in diagnosis of Alzheimer disease.

[Y-specific sequences in Turner syndrome]. / Oznaczanie swoistych sekwencji chromosomu Y w zespole Turnera.

Turner Syndrome (TS) is the only one monosomy that occurrs+ in humans. The cytogenetics of TS is very well known from years. It has been estimated that almost 98-99% of TS foetuses end in abortion. It was suggested that the monosomy arises relatively late during embryonal development and survived TS individuals could be mosaics. It has been proved that mosaic karyotype mos 45,X/46X, + mar(Y) occurrs++ in 2% to 11% of TS patients. The patients having additional cell line containing der(Y) are at increased risk of gonadoblastoma development. In these cases gonadectomy should be considered. Therefore detection of mosaic and establishing the origin of marker chromosome (specially containing Y-specific sequences) is of special importance. The aim of present study was to detect the small mosaics, containing mar(Y) in TS patients, by using PCR and FISH techniques. Eight Y sequences for the PCR analyses as well as bicolor in situ hybridisation with painting probes for Y and X chromosomes have been applied. The positive amplification for Y-specific sequences has been detected in 7% of TS patients. Our results support the thesis that searching for the Y sequences should be introduced to routine genetic TS diagnosis.

Variable distribution of TFF2 (Spasmolysin) alleles in Europeans does not indicate predisposition to gastric cancer.

Peptides belonging to the trefoil factor family (TFF) protect the gastrointestinal epithelia. Overexpression of TFFs was observed in pathological conditions such as gastritis, ulceration, metaplasia and neoplasia of the gastrointestinal tract. The aims of this work were to investigate the recently described TFF2 gene polymorphism in different European populations. DNA samples from blood of healthy individuals and gastric cancer patients were genotyped using the polymerase chain reaction. They were compared to a gastric cancer population. The results do not show any significant difference in allelic frequencies between gastric cancer patients and healthy individuals from Portugal. However, the frequency of the two alleles found varies considerably among Europeans.

Determination of phosphoglucomutase phenotypes in hair bulbs.

The research was to determine a simple method of phosphoglucomutase phenotype identification in hair bulb. The agarose technique and electrophoresis on cellulose-acetate foil methods were chosen because a small quantity of the maternal available for examination. It was found out that 1 or 2 bulbs are sufficient to identify the PGM1 features if the electrophoresis method is applied and if more bulbs are available, the PGM3 characteristic can also be identified. The modified technique was used for staining the phosphoglucomutase phenotypes.

[The "blotting" method for the detection of possible phenotypes of acid phosphatase in sperm]. / Das "Blotting" Verfahren zum Nachweis möglicher Phänotypen der Sauren Phosphatase des Spermas.

64 sperm samples are tested for the evidence of Sperm Acid Phosphatase by agarose isoelectrofocusing with modified blotting. Near pI = 5.0, we observed polymorphic fractions of Sperm Acid Phosphatase. The investigated polymorphism is alike the original one from prostata. Polymorphic patterns are stable in sperm stains up to 6 weeks.

Polymorphism of acid phosphatase (E.C. 3.1.3.2) including some rare variants in Wroclaw (south-west Poland) area. Utility in population and criminalistic studies.

In a sample of 7467 inhabitants of Wroclaw and Lower Silesia, the frequencies of ACP alleles were determined as ACPa = 0.3466. ACPb = 0.5684, ACPc = 0.0848, ACPr = 0.001. The above frequencies are similar to those described in other population samples. Phenotypes with rare gene ACPe were described in Poland for the first time. Utility of ACP phenotyping in criminalistical investigation of legal instruments is strongly limited by low stability of enzyme and change of electrophoretic image.

Genetic polymorphism of FUC (EC 3.2.1.51.) in Polish population.

In a sample of the Polish population numbering 271 persons three FUC phenotypes were encountered by cellulose acetate gel isoelectric focusing (CAGIF). The frequencies of FUC1 and FUC2 genes were 0.653 and 0.347, respectively. The FUC system has proven to be of high value for paternity testing.

Rare phenotypes of the phosphoglucomutase locus 1 detectable by isoelectric focusing on Cellogel.

The rare phenotypes PGM1, determined by alleles PGM1(3), PGM1(4), PGM1(6), and PGM1(7) were examined by starch gel electrophoresis and cellulose acetate gel isoelectric focusing and were compared with the commonest phenotypes of PGM1. The frequencies of the rare genes found in the Polish populations were as follows: in Lublin, PGM1(3) = 0.0002, PGM1(4) = 0.0005, PGM1(6) = 0.0010, and PGM1(7) = 0.0005; in Wroclaw, PGM1(3) = 0.0000, PGM1(4) = 0.0005, PGM1(6) = 0.0007, and PGM1(7) = 0.0002. The results suggest that the F and S type variants of the genes PGM1(4) and PGM1(7) probably do not occur. It is still possible that F and S variants exist for the genes PGM1(3) and PGM1(6).

Subtypes of the phosphoglucomutase-1 (PGM1) locus detectable in Polish populations by isoelectric focusing on cellogel.

The technique of isoelectric focusing on methylated 'cellogel' strips (CAGIF) was used to confirm the presence of four alleles of PGM1 in human red cell lysates. The subtypes of PGM1 were determined in two Polish population samples, from Southwestern Poland (Wroclaw region, n=321) and Southeastern Poland (Lubin region, n=212). Ten different phenotypes are considered as gene products of four alleles at PGM1, with the following frequencies: Wroclaw: PGM1F, 0.1044; PGM1S, 0.5966; PGM2F, 0.0685; and PGM2S, 0.2305; Lublin: PGM1F, 0.1439; PGM1S, 0.6014; PGM2F, 0.0825; and PGM2S, 0.1722.

GLO polymorphism in two Polish population samples.

The red cell GLO phenotypes were determined in two Polish population samples. A total of 1310 people from the region of Lublin (Southeastern Poland, n = 797) and Wroclaw (Southwestern Poland, n = 513) were investigated. The gene frequencies were calculated for GLO1 (= 0.4427) and GLO2 (= 0.5573). The evaluation of 372 mother-child pairs showed no deviation from a hereditary hypothesis.