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Cell synchronization was used to obtain an adequate percentage of very long chromosomes in equine mitotic spreads. Reported here is our variation, adapted to horse chromosomes, of a method using excess thymidine followed by bromodeoxyuridine incorporation. This technique routinely yields excellent quality cells, predominantly in prometaphase and prophase. Among other differences with the standard technique, this addltion does not use Colcemid, which, in addltion to Inhibiting spindle fiber formation, also increases chromosome contraction resulting in thicker and thus fewer bands. Consequently, horse prometaphase chromosomes, which have incorporated BrdU in the late-S-phase, are very long and display a large number of R-bands after the fluorescence-photolysis Glemsa method. This technique should definitely be useful for the analysis of structural anomalies and the standardization of equine R-bands

Both dynamic G-banding and cell synchronization produced by bromodeoxyuridine (BrdU), were applied to equine chromosomes. BrdU incorporated during the first half of the S-phase is taken up into the R-bands that are early replicating. These bands, which have incorporated BrdU, cannot contract as usual and remain elongated; only the other regions of the chromosome, i.e., the G-bands, contract normally and are sharply defined. BrdU also can be used for cell synchronization. The addition of BrdU in a high concentration, 15 hours before harvest, and its removal 11 hours later, has two effects: initially the BrdU is incorporated during the first part of the S-phase and then it blocks the cells at mid-S-phase. Within the cell cycle, mid-S-phase appears to be the most vuinerable time to various blocking agents. To differentiate the regions of BrdU incorporation from those that have not been substituted, the fluorescence-photolysis-Glemsa (FPG) technique was applied as modified for horse chromosomes. This dynamic technique, which produces many prometaphase and prophase chromosomes showing very sharp G-bands, is certain to enhance the accuracy of cytogenetic analysis and aid in the standardization of equine chromosomes

It is common in archeology and some forensic cases to make assumptions about the relationship between persons buried in the same grave, or in distinct burials but in the same cemetery. However, there is no objective criterion to make such suppositions. This study aims to investigate the biological relationship among six Neolithic individuals (4200–3400 cal BC) from the Can Gambús-1 necropolis (Barcelona, Spain) as well as, their possible mitochondrial ancestry. From at least 47 tombs, six skeletons were selected, from six individual graves, selecting from each one three integral teeth, without cavities or caries. To carry out this study, the samples were analyzed by two distinct extraction methods, one destructive and other non-destructive, and to determine a presumptive (biological) kinship between the buried individuals, small length nuclear DNA markers were considered, as well as a mitochondrial DNA analysis, in order to determine their maternal lineage. The samples were extremely degraded, requiring several attempts to achieve sequences with a standard quality, without double peaks or other artefacts. Despite the degraded state of the samples, it has been possible to analyze the mitochondrial information and to determine that the studied individuals had a European ancestry, and were not maternally related, since they did not share the same mitochondrial haplotype.

The nature of the sample in a forensic case is one of the most important factors, since it determines the posterior analysis, helping to define or discard its identity (like blood versus semen). A presumptive test is a qualitative analysis that allows to identify, or confirm, the presence of a substance in a sample. These determinations usually occur, after a chemical reaction, and a specific colour is produced. A false positive is another substance reacting the same way, producing the expected result. The aim of this work was to evaluate the most effective presumptive test (with fewer false-positives) when analysing products that could look and behave like blood during a forensic screening assay. Eight different products were tested, like Betadine®, and four reagents were considered: Tetramethylbenzidine, O-toluidine, Leuchomalachite green and BlueStar® Forensic (BlueStar). Each product was tested with the reagents five times − mixed with human blood (3:1), with three different animal blood (3:1), and then unmixed. Our results indicated that Leuchomalachite green is the most suitable presumptive test, since it was the reagent with less false positives.

One of the most challenging points for ancient DNA studies on human remains is that analysing procedures involve partial or total destruction of the samples, which usually are unique and irreplaceable. This is what prevents museums, anthropologists or archaeologists from giving samples for genetic investigation. So that it is interesting to find an analysing method without destroying them. In this study, it was carried out the evaluation of a non-destructive extraction DNA protocol on 4 Neolithic individuals, from Can Gambús (Sabadell, Spain) and 4 pre Bell Beaker period individuals, from Los Cercados (Valladolid, Spain). Along this work, it was discussed the efficiency of the non-destructive protocol, valuing the obtained result, and the evidences state of preservation after the whole procedure.

On their arrival to the Iberian Peninsula, the North African and Berbers Muslims encountered no resistance, but a land with some political trouble between Christians, Godos and the Jewish people, which facilitated somehow the absorption of the new invaders ideas. In on hand, there is the idea that military invasion changed by force daily habits, religious traditions and beliefs: the “Al-Ândalus” region. However, in the other hand, there are evidences of a peaceful co-existence between Muslims and the “Iberians”, as well as, a voluntary absorption of the Islamic culture. Thus, Muslims who lived under Christian rule formed the “Mudejar” community and its biogeographical origin remains still unclear. Considering the wide geographical extend and duration of the Muslim occupation, it is reasonable to suppose that the Islamic permanency had a significant impact on the Iberian gene-pool. In Uceda, (Guadalajara, Spain), 70 medieval bodies were found buried in a Mudejar cemetery. According to anthropologists’ experts, Muslim burial was observed, including the body orientation to Mecca, and all led to believe that the individuals had an Islamic origin. So, we are carrying out a genetic study in order to verify if their mitochondrial lineage is coincident with the characteristic haplogroups described for peoples from the actual Islamic geographical areas. The sampling process concerned two teeth per individual, obeying to the criteria to select evidences for critical DNA analysis. The samples were directly exposed to the floor conditions, and, in some cases, were externally damaged, requiring some adaptations in the analysis process. Our preliminary results seem to indicate that the individuals did not have North-African origin, since they revealed, so far, European haplogroups, indicating a possible religious conversion. From the forensic perspective, this is an important study since we adapted the technique in order to obtain reliable and replicable data from critical sample

There has been very little work published on the variation of reporting practices of mixtures between laboratories, but it has been previously demonstrated that there is little consistency. This is because there is no current uniformity of practice, so different laboratories will operate using different rules. The interpretation of mixtures is not solely a matter of using some software to provide ‘an answer’. An assessment of a case will usually begin with a consideration of the circumstances of a crime. Assumptions made about the numbers of contributors follow from an examination of the electropherogram(s) – and these may differ between the prosecution and the defence hypotheses. There may be a necessity to evaluate several sets of hypotheses for any given case if the circumstances are uncertain. Once the hypotheses are formulated, the mathematical analysis is complex and can only be accomplished by the use of specialist software. In order to obtain meaningful results, it is essential that scientists are trained, not only in the use of the software, but also in the methodology to understand the likelihood ratio concept that is used. The Euroforgen-NoE initiative has developed a training course that utilizes the LRmix program to carry out the calculations. This software encompasses the recommendations of the ISFG DNA commissions on mixture interpretation and is able to interpret samples that may come from two or more contributors and may also be partial profiles. Recently, eighteen different laboratories were trained in the methodology. Afterwards they were asked to independently analyze two different cases with partial mixture DNA evidence and to write a statement court-report. We show that by introducing a structured training programme, it is possible to demonstrate, for the first time, that a high degree of standardization, leading to uniformity of results can be achieved by participating laboratories., There has been very little work published on the variation of reporting practices of mixtures between laboratories, but it has been previously demonstrated that there is little consistency. This is because there is no current uniformity of practice, so different laboratories will operate using different rules. The interpretation of mixtures is not solely a matter of using some software to provide 'an answer'. An assessment of a case will usually begin with a consideration of the circumstances of a crime. Assumptions made about the numbers of contributors follow from an examination of the electropherogram(s)--and these may differ between the prosecution and the defence hypotheses. There may be a necessity to evaluate several sets of hypotheses for any given case if the circumstances are uncertain. Once the hypotheses are formulated, the mathematical analysis is complex and can only be accomplished by the use of specialist software. In order to obtain meaningful results, it is essential that scientists are trained, not only in the use of the software, but also in the methodology to understand the likelihood ratio concept that is used. The Euroforgen-NoE initiative has developed a training course that utilizes the LRmix program to carry out the calculations. This software encompasses the recommendations of the ISFG DNA commissions on mixture interpretation and is able to interpret samples that may come from two or more contributors and may also be partial profiles. Recently, eighteen different laboratories were trained in the methodology. Afterwards they were asked to independently analyze two different cases with partial mixture DNA evidence and to write a statement court-report. We show that by introducing a structured training programme, it is possible to demonstrate, for the first time, that a high degree of standardization, leading to uniformity of results can be achieved by participating laboratories.

Tissue kallikrein (KLK1) is a kinin-forming serine protease synthesized in many organs including arteries and kidney. Study of the physiological role of KLK1 has benefited from the availability of mouse and human genetic models of KLK1 deficiency, through engineering of KLK1 mouse mutants and discovery of a major polymorphism in the human KLK1 gene that induces a loss of enzyme activity. Studies in KLK1-deficient mice and human subjects partially deficient in KLK1 have documented its critical role in arterial function in both species. KLK1 is also involved in the control of ionic transport in the renal tubule, an action that may not be kinin-mediated. Studies of experimental diseases in KLK1-deficient mice have revealed cardio- and nephro-protective effects of KLK1 and kinins in acute cardiac ischemia, post-ischemic heart failure, and diabetes. Potential clinical and therapeutic developments are discussed.