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In contrast to members of Picornaviridae which have long 5′-untranslated regions (5′UTRs) containing internal ribosomal entry sites (IRESs) that form five distinct classes, members of Caliciviridae typically have short 5′UTRs and initiation of translation on them is mediated by interaction of the viral 5′-terminal genome-linked protein (VPg) with subunits of eIF4F rather than by an IRES. The recent description of calicivirus genomes with 500–900nt long 5′UTRs was therefore unexpected and prompted us to examine them in detail. Sequence analysis and structural modelling of the atypically long 5′UTRs of Caliciviridae sp. isolate yc-13 and six other caliciviruses suggested that they contain picornavirus-like type 2 IRESs, whereas ruddy turnstone calicivirus (RTCV) and Caliciviridae sp. isolate hwf182cal1 calicivirus contain type 4 and type 5 IRESs, respectively. The suggestion that initiation on RTCV mRNA occurs by the type 4 IRES mechanism was confirmed experimentally using in vitro reconstitution. The high sequence identity between identified calicivirus IRESs and specific picornavirus IRESs suggests a common evolutionary origin. These calicivirus IRESs occur in a single phylogenetic branch of Caliciviridae and were likely acquired by horizontal gene transfer.

Initiation of translation on many viral mRNAs occurs by non-canonical mechanisms that involve 5’ end-independent binding of ribosomes to an internal ribosome entry site (IRES). The ~190 nt-long intergenic region (IGR) IRES of dicistroviruses such as Cricket paralysis virus (CrPV) initiates translation without Met-tRNAiMetor initiation factors. Advances in metagenomics have revealed numerous dicistrovirus-like genomes with shorter, structurally distinct IGRs, such as Nedicistrovirus (NediV) and Antarctic picorna-like virus 1 (APLV1). Like canonical IGR IRESs, the ~165 nt-long NediV-like IGRs comprise three domains, but they lack key canonical motifs, including L1.1a/L1.1b loops (which bind to the L1 stalk of the ribosomal 60S subunit) and the apex of stemloop V (SLV) (which binds to the head of the 40S subunit). Domain 2 consists of a compact, highly conserved pseudoknot (PKIII) from which a CrPV-like stemloop SLIV and an adjacent conserved 5nt-long stemloop protrude.In vitroreconstitution experiments showed that NediV-like IRESs initiate translation from a non-AUG codon and form elongation-competent 80S ribosomal complexes in the absence of initiation factors and Met-tRNAiMet. Unlike canonical IGR IRESs, NediV-like IRESs bind directly to the peptidyl (P) site of ribosomes leaving the aminoacyl (A) site accessible for decoding. The related structures of NediV-like IRESs and their common mechanism of action indicate that they exemplify a distinct class of IGR IRES.

Utilization of non-AUG alternative translation start sites is most common in bacteria and viruses, but it has been also reported in other organisms. This phenomenon increases proteome complexity by allowing expression of multiple protein isoforms from a single gene. In Saccharomyces cerevisiae, a few described cases concern proteins that are translated from upstream near-cognate start codons as N-terminally extended variants that localize to mitochondria. Using bioinformatics tools, we provide compelling evidence that in yeast the potential for producing alternative protein isoforms by non-AUG translation initiation is much more prevalent than previously anticipated and may apply to as many as a few thousand proteins. Several hundreds of candidates are predicted to gain a mitochondrial targeting signal (MTS), generating an unrecognized pool of mitochondrial proteins. We confirmed mitochondrial localization of a subset of proteins previously not identified as mitochondrial, whose standard forms do not carry an MTS. Our data highlight the potential of non-canonical translation initiation in expanding the capacity of the mitochondrial proteome and possibly also other cellular features.

Utilization of non-AUG alternative translation start sites is most common in bacteria and viruses, but it has been also reported in other organisms. This phenomenon increases proteome complexity by allowing expression of multiple protein isoforms from a single gene. In Saccharomyces cerevisiae, a few described cases concern proteins that are translated from upstream near-cognate start codons as N-terminally extended variants that localize to mitochondria. Using bioinformatics tools, we provide compelling evidence that in yeast the potential for producing alternative protein isoforms by non-AUG translation initiation is much more prevalent than previously anticipated and may apply to as many as a few thousand proteins. Several hundreds of candidates are predicted to gain a mitochondrial targeting signal (MTS), generating an unrecognized pool of mitochondrial proteins. We confirmed mitochondrial localization of a subset of proteins previously not identified as mitochondrial, whose standard forms do not carry an MTS. Our data highlight the potential of non-canonical translation initiation in expanding the capacity of the mitochondrial proteome and possibly also other cellular features.

The allele frequencies of 15 autosomal STR loci (D3S1358, vWA, FGA, TH01, TPOX, CSF1PO, D5S818, D13S317, D7S820, D16S539, D2S1338, D8S1179, D21S11, D18S51, and D19S433) used in forensic medicine were determined for the Russian population of European Russia (N = 176). The power of discrimination (PD) and power of exclusion (PE) of the system of the 15 STR loci were 0.999 999 999 999 999 986 and 0.999 999 331 310 171 000, respectively. The allele and genotype frequency distributions in the Russian population corresponded to the Hardy-Weinberg equilibrium. The D2S1338, D18S51, D21S11, and FGA loci were identified as the most informative markers for the Russian population and proposed as a reference for forensic studies in the Russian Federation.

We have obtained Y-STR haplotypes in 12 loci (DYS19, DYS385, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438 and DYS439) from 215 Buryat males. We have found that one haplotype (15-11,18-13-28-23-10-11-14-14-10-12) comprises more than 30% of Y chromosomes in this population while another haplotype (14-11,13-14-30-23-10-14-14-14-10-10) comprises additional 14% of chromosomes. The population under study seems to be very homogenous as far as Y chromosome is regarded and the most frequent haplotype seems to be the modal haplotype for Buryats.

Many well-defined mutations in the gene for the catalytic subunit of polymerase gamma (POLG1) have been found to be associated with disease, whereas the status of several mutations remains unresolved due to the conflicting reports on their frequencies in populations of healthy individuals. Here, we have developed a highly sensitive, real-time allelic discrimination assay enabling detection of the Y831C mutation in the POLG1 gene. The Y831C mutation is present in the Polish population at a frequency of 2.25%. The new assay is well suited to both extensive population studies and molecular diagnostics of POLG1.

In order to investigate the genetic history of autochthonous South Siberian populations and to estimate the contribution of distinct patrilineages to their gene pools, we have analyzed 17 Y-chromosomal binary markers (YAP, RPS4Y(711), SRY-8299, M89, M201, M52, M170, 12f2, M9, M20, 92R7, SRY-1532, DYS199, M173, M17, Tat, and LLY22 g) in a total sample of 1,358 males from 14 ethnic groups of Siberia (Altaians-Kizhi, Teleuts, Shors, Tuvinians, Todjins, Tofalars, Sojots, Khakassians, Buryats, Evenks), Central/Eastern Asia (Mongolians and Koreans) and Eastern Europe (Kalmyks and Russians). Based on both, the distribution pattern of Y-chromosomal haplogroups and results on AMOVA analysis we observed the statistically significant genetic differentiation between the populations of Baikal and Altai-Sayan regions. We suggest that these regional differences can be best explained by different contribution of Central/Eastern Asian and Eastern European paternal lineages into gene pools of modern South Siberians. The population of the Baikal region demonstrates the prevalence of Central/Eastern Asian lineages, whereas in the populations of Altai and Sayan regions the highest paternal contribution resulted from Eastern European descent is revealed. Yet, our data on Y-chromosome STRs variation demonstrate the clear differences between the South Siberian and Eastern European R1a1-lineages with the evolutionary ages compatible with divergence time between these two regional groups.

Summary To investigate the origin and evolution of aboriginal populations of South Siberia, a comprehensive mitochondrial DNA (mtDNA) analysis (HVR1 sequencing combined with RFLP typing) of 480 individuals, representing seven Altaic-speaking populations (Altaians, Khakassians, Buryats, Sojots, Tuvinians, Todjins and Tofalars), was performed. Additionally, HVR2 sequence information was obtained for 110 Altaians, providing, in particular, some novel details of the East Asian mtDNA phylogeny. The total sample revealed 81% East Asian (M ∗ , M7, M8, M9, M10, C, D, G, Z, A, B, F, N9a, Y) and 17% West Eurasian (H, U, J, T, I, N1a, X) matrilineal genetic contribution, but with regional differences within South Siberia. The highest influx of West Eurasian mtDNAs was observed in populations from the East Sayan and Altai regions (from 12.5% to 34.5%), whereas in populations from the Baikal region this contribution was markedly lower (less than 10%). The considerable substructure within South Siberian haplogroups B, F, and G, together with the high degree of haplogroup C and D diversity revealed there, allows us to conclude that South Siberians carry the genetic imprint of early-colonization phase of Eurasia. Statistical analyses revealed that South Siberian populations contain high levels of mtDNA diversity and high heterogeneity of mtDNA sequences among populations (Fst = 5.05%) that might be due to geography but not due to language and anthropological features.

We have performed haplotyping of seven Y-STRs (DYS19, DYS389, DYS390, DYS391, DYS392, DYS393 and DYS385). We have found 199 different haplotypes in 277 unrelated males from Pomerania–Kujawy region of Poland. Among the haplotypes found, 164 (59.2%) were unique in our population sample, while 35 occurred more than once. Interpopulation comparisons were also performed and showed significant differences in haplotype frequencies between our population and other populations of Western and Southern European origin, while haplotype distributions in populations from other regions of Poland, as well as Central and Eastern Europe, were similar.

The authors would like to thank the following individuals for providing samples for this study: Drs. E. Lotosh, F. Luzina, I. Dambueva, C. Dorzhu, V. Kakpakov, and O. Karamchakova. This work was supported by the Russian Fund for Basic Research (grant 99-06-80430), the State Program Frontiers in Genetics (grant 99-04-30), and by a grant from the Ludwik Rydygier Medical University in Bydgoszcz (BW 90/01).

Allele frequencies for 13 STRs were obtained from a sample of 306-1041 unrelated individuals born in the Pomerania-Kujawy region of Poland.

Single-strand conformation polymorphism (SSCP) analysis combined with automated laser fluorescence detection is proposed as a comprehensive, rapid and sensitive method for screening sequence variation of the human beta-actin-related pseudogene (HUMACTBP2). Eleven sequenced alleles representing each type of known sequence variant of HUMACTBP2 locus were studied. Allelic variants of the same size but different sequence structures are easily resolved on the basis of their secondary conformation. Fifty ACTBP2 amplification products previously typed on a denaturing gel were repeatedly examined to determine the utility of SSCP analysis in terms of ease of interpretation and reproduction capabilities of the conformational patterns. Eleven sequenced ACTBP2 allelic variants were used as external conformation standards in polymerase chain reaction (PCR)-SSCP subtyping. This enabled identification of polymorphism in a particular length variant and therefore consistent discrimination between heterozygous samples appeared identical on denaturing gels. Of five "homozygous" samples, one was shown to be heterozygous for two distinct alleles of the same size but different sequences. Thus, the method provides a unique possibility for detecting false homozygotes. The technique complements both denaturing gel electrophoresis and DNA sequencing in studies on the overall variability of the ACTBP2 locus.

An automated DNA profiling system based on the multiplex amplification of highly polymorphic short tandem repeat (STR) markers and the amelogenin locus was developed. Five STR loci with nonoverlapping allele size ranges have been utilized in the multiplex amplifications, including HUMD1S103, HUMTH01, HUMD21S11, HUMD18S51, and HUMFIBRA. One primer for each locus was labeled with a fluorescent dye (fluorescein) which allows detection on the single wavelength ALF DNA Sequencer (Pharmacia Biotech). As part of the detailed evaluation of the suitability of the hexaplex system for routine forensic use, the effect of variation in amplification parameters on the efficiency of the system was examined. Polymerase chain reaction amplification conditions were optimized to provide specific, robust amplification of forensic samples.

IgA nephropathy is currently the most frequently occurring type of primary glomerulonephritis. Studies aimed at determining the factors of favorable and unfavorable prognosis in the progression of the disease are conducted. Apart from the above renal disease progression factors, it seems that renal functional reserve (RFR), indirectly indicating the functional status of intrarenal vessels can be a marker assisting in determining prognosis and effectiveness of applied treatment. Decrease in RFR is one of the first symptoms of renal damage, since it precedes decrease in GFR assessed in resting condition. The aim of our study was to assess selected functional (RFR), metabolic, and genetic parameters of renal disease progression in patients with IgA nephropathy, as well as to determine their effect on clinical progression of the disease. Material and methods. The study comprised 30 patients with renal biopsy proven IgA nephropathy, aged 35,2 +/- 8,9, 12 women and 18 men, who had conducted a 12-month period of observation and treatment. The patients' RFR was measured and the following parameters in blood pressure samples were established: creatinine, BUN, uric acid, total cholesterol (TCH), HDL and LDL and TG, homocysteine, endothelium functional indicators: vWF:Ag, TPA:Ag, PAI-1, polymorphism of the human angiotensin converting enzyme gene and endothelial nitric oxide synthase gene. In 24-hour urine collection N-acetylglucosaminidase excretion and daily protein loss were measured. Results. During treatment, changes in some biochemical indicators were observed (uric acid, TCH, LDL, DUB, NAG, erythrocyturia, homocysteine), while others remained stable. Statistically significant differences in concentrations of endothelial antigens: vWF:Ag and PAI-1 were found. Conclusions. Based on the analysis of the results it was concluded that functional status of intrarenal vessels is related to functional status of endothelium and renal tubulae, and also that it probably affects the response to treatment. Decrease of proteinuria during treatment is, among others, related to decrease of metabolic disorders, while the initial results of analysis of polymorphism of the human angiotensin converting enzyme gene and endothelial nitric oxide synthase gene suggest that it may affect the decrease of proteinuria and concentration of homocysteine in the blood.

The role of Helicobacter pylori (H. pylori) infection in etiopathogenesis of gastritis and/or duodenitis is well recognized and proven. H. pylori infection also concerns children and youth. It was proven that CagA(+) strains of H. pylori cause inflammatory changes and can lead to ulcer disease of ventriculum and/or duodenum.The purpose of the study was to determine the prevalence of urease A and cytotoxicity genes in children with diagnosed H. pylori infection and concomitant gastritis and/or duodenitis as well as to determine the kind and stadium of inflammatory changes in relation to particular strains.The study was performed in 76 patients aged from 3 to 20 years (average age 12.6), who were qualified for endoscopy of the upper GI tract because of recurrent abdominal pain complaints. The H. pylori infection and accompanying gastritis and/or duodenitis were diagnosed according to the Sydney classification.In 57/76 (75%) of diagnosed patients the H. pylori infection was proven with the PCR method (ureA (+)). The following strains were found: CagA(+) in 19 (33% infected subjects). Gastritis and/or duodenitis occurred in all subjects infected with CagA (+) strains and in 54/57 of those with H. pylori infection ureA (+). In subjects without infection, gastritis and/or duodenitis was found in 12 persons, and this result was significantly lower.In diagnosis it is important to confirm H. pylori presence by CagA gene typing because it relates to the inflammatory changes of the stomach and/or duodenum.

Summary Mitochondrial DNA variability in the Polish Roma population has been studied by means of hypervariable segment I and II (HVS I and II) sequencing and restriction fragment-length polymorphism analysis of the mtDNA coding region. The mtDNA haplotypes detected in the Polish Roma fall into the common Eurasian mitochondrial haplogroups (H, U3, K, J1, X, I, W, and M*). The results of complete mtDNA sequencing clearly indicate that the Romani M*-lineage belongs to the Indian-specific haplogroup M5, which is characterized by three transitions in the coding region, at sites 12477, 3921 and 709. Molecular variance analysis inferred from mtDNA data reveals that genetic distances between the Roma groups are considerably larger than those between the surrounding European populations. Also, there are significant differences between the Bulgarian Roma (Balkan and Vlax groups) and West European Roma (Polish, Lithuanian and Spanish groups). Comparative analysis of mtDNA haplotypes in the Roma populations shows that different haplotypes appear to demonstrate impressive founder effects: M5 and H (16261–16304) in all Romani groups; U3, I and J1 in some Romani groups. Interestingly, haplogroup K (with HVS I motif 16224-16234-16311) found in the Polish Roma sample seems to be specific for Ashkenazi Jewish populations.

Allele frequencies for 15 short tandem repeats (STRs) included in AmpFlSTR Profiler and AmpFlSTR SGM Plus kits were obtained from populations of Pomorze Gdanskie, Wielkopolska, Kujawy, Pomorze Zachodnie, Mazury and Mazowsze regions of Poland.

Mitochondrial DNA (mtDNA) sequence variation was examined in Poles (from the Pomerania-Kujawy region; n = 436) and Russians (from three different regions of the European part of Russia; n = 201), for which the two hypervariable segments (HVS I and HVS II) and haplogroup-specific coding region sites were analyzed. The use of mtDNA coding region RFLP analysis made it possible to distinguish parallel mutations that occurred at particular sites in the HVS I and II regions during mtDNA evolution. In total, parallel mutations were identified at 73 nucleotide sites in HVS I (17.8%) and 31 sites in HVS II (7.73%). The classification of mitochondrial haplotypes revealed the presence of all major European haplogroups, which were characterized by similar patterns of distribution in Poles and Russians. An analysis of the distribution of the control region haplotypes did not reveal any specific combinations of unique mtDNA haplotypes and their subclusters that clearly distinguish both Poles and Russians from the neighbouring European populations. The only exception is a novel subcluster U4a within subhaplogroup U4, defined by a diagnostic mutation at nucleotide position 310 in HVS II. This subcluster was found in common predominantly between Poles and Russians (at a frequency of 2.3% and 2.0%, respectively) and may therefore have a central-eastern European origin.

The work presents 3 members of a family of 4, who were diagnosed to have coeliac disease (one classic and two latent forms of the disease). Genetic investigation regarding all three patients revealed the existence of HLA DQ A1*0501 allele associated with susceptibility to coeliac disease. Due to a much more frequent occurrence of atypical forms of coeliac disease in family members, than in general population, and due to risks resulting from tardy diagnoses and the lack of treatment, it is recommended that patients should be subjected to tests determining the presence of antiendomysial antibodies, as well as to genetic investigation with regards to latent coeliac disease.

This paper presents the results of a Polish pop- ulation study (n = 210) for the three STR loci vWA, D3S1358 and FGA analysed using the multiplex PCR system AmpflSTR Blue. The allele distributions were in accordance with Hardy-Weinberg expectations. The com- bined mean exclusion chance, mean paternity index and power of discrimination for the three loci were MEC = 0.96055, MPI = 127.1295 and PD = 0.99986. This demonstrates that these systems are valuable tools for forensic identification and paternity testing.

The human beta-actin related pseudogene (HUMACTBP2) seems to be one of the most informative microsatellite markers known because of the high number of length and sequence variants. A total of 50 alleles found in white Caucasians from the Pomerania-Kujawy region of Poland were analyzed by automated sequencing. In addition to STR length polymorphism, seven different types of sequence variation were observed. Alleles ranging in size between 233 and 273 bp showed regular sequence structure with tetranucleotide repeats AAAG. In the alleles ranging in size from 275 to 323 bp, hexamer units AAAAAG or AGAAAG occurred in the repeat region in addition to AAAG repeats. Two alleles (317 and 321 bp) contained two hexamers in the repeat region. There was considerable polymorphism of the hexamer position leading to allelic variants of the same size but different sequence structures. A large amount of variation in both 5' and 3' flanking regions was also observed. Allelic designation based on the number of all types of units within the repeat region (including the hexamer unit) is proposed. An allelic ladder composed of 21 sequenced alleles was constructed to add precision and accuracy to the identification of alleles at ACTBP2 locus.

The application of any new genetic system in forensic casework requires a sufficiently large data base of population. In the present study, we report the allele frequency distribution for seven different loci in the Polish population of Pomerania-Kujawy Region. Five of these markers are coamplified simultaneously using the Amplitype PM kit (Perkin-Elmer): low density lipoprotein receptor (LDLR), glycophorin A (GYPA), hemoglobin G gammaglobin (HBGG), D7S8 linked to cystic fibrosis locus and Group specific Component (GC). The other two loci are VNTR D1S80 (Nakamura 1988) and STR ACTBP2 (SE33) (Polymeropoulos 1992). In addition, we have sequenced selected SE 33 alleles to determine possible sequence structure variations which may cause the allele designation difficult.

Before the developement of DNA polymorphism the most cases of disputed paternity were solved by testing blood groups, serum proteins, erythrocyte enzymes and additionally HLA. Nevertheless in cases of only one system exclusion, especially homozygotic ones, or silent genes the further extension of surveys, family investigations or finally investigation of the enzyme activity has been required. Those cases have been suggested by some authors to be resolved using the DNA technique [4,7,8,10]. Since Nakamura et al. [9] had reported the polymorphic region — VNTR locus D1S80 and Kasai et al. [3] had sequenced it the new approach caused by the introduction of D1S80 into disputed paternity investigation has been used.

The D1S80 locus contains repetitive DNA sequences that are arranged as tandem repeat units (VNTR). The D1S80 tandem repeat unit defined by Nakamura et al. [4] is 16 base pairs in length repeated from 14 to over 41 times for each allele. The D1S80 locus has been localized to the distal end of chromosome 1p and shows codominant autosomal segregation [1–5].

Most methods of clinical diagnostics, such as different radiological, ultrasonographic or finally endoscopie examinations have very limited use in forensic medicine. It is the result of the fact, that a pathologist can evaluate almost everything directly, with no application of any device. There are only a few published papers concerning the forensic autopsies assisted with such methods [1–7].

Using post mortem temperature for estimating the time since death began in last century The first really comprehensive paper is dated 1863 [1]. Since then, over one hundred papers on this topic have been published. Many possible measurement points were investigated in detail [2, 3]. Several algorithms are known, based either on purely practical measurements or on theoretical considerations tested by experiment. Some of them are just rules of thumb, easy to memorize and use, whereas others require a bit more advanced mathematics [4–6], even involving computer data processing in some cases [9–12].

Chromatographic conditions for the separation of five pairs of active compounds occuring in four ointment preparations and an aerosol one were elaborated. The ways of extraction of hydrocortisone esters (acetate and butyrate), chlorquinaldol, oxytetracycline base and oxytetracycline hydrochloride from the ointments as well as purification of the extracts prior to the HPLC analysis were described. The proposed analytical methods are for more specific and precise from those used until now in the home-made preparations quality control.