A new search tool for familial searches in the Israel Criminal DNA database.

The Israel DNA database has recently started to conduct familial searches (FS). We adopted the CODIS pedigree strategy for FS, which is used in the Unidentified Human Remains (UHR) database and implemented it into the criminal forensic database. This strategy is based on Kinship analysis performed in pedigrees containing DNA profiles from the crime scene designated "unknown," that are then searched against the entire suspects database. A list of candidates is produced and ranked by Joint Pedigree Likelihood Ratio (JPLR). Traditional Y-STR characterizing and mitochondrial sequencing can be performed in order to further minimize the list. Our novel strategy consists of an additional pedigree analysis aimed at prioritizing potential candidates from the candidate list: a Test Pedigree Tree (TPT). Candidates ranked high on the JPLR list can be verified or eliminated from the list by using other close family members included in the database. To further validate this novel strategy, we describe two cases where implementation of this strategy led to a successful match and solved the crime.

Weight-of-evidence for DNA identification of missing persons and human remains using CODIS.

DNA Identification of unidentified human remains (UHR) is performed in Israel by comparing the UHR's short tandem repeat (STR) profiles to a national database of STR profiles taken from relatives of missing persons. Kinship analysis is performed using the CODIS 7.0 software and results are stated as a Joint Pedigree Likelihood Ratio (JPLR). The weight-of-evidence for JPLR has never been studied, making it difficult to interpret the meaning of specific values in terms of whether UHR are related to specific pedigrees. Therefore, the aim of this study was to statistically determine the practical meaning and context of the JPLR. We used 440 million pairs of simulated DNA profiles and 294 pairs of real ones from known siblings, parent/offspring and unrelated persons. A Score-Based Likelihood Ratio (SBLR) was empirically constructed, validated and compared to both JPLR and the LR produced by CODIS. Our results show that CODIS's JPLR and LR values for single-person pedigrees overestimate the level of support for both "parent/child" and "siblings" propositions relative to the "unrelated" proposition, by up to two orders of magnitude. A practical table is given for correcting this phenomenon, with statistical interpretation (i.e. SBLR) for each JPLR score, including verbal levels of propositional support ranging from "no support" (SBLR<2) to "extremely strong" (SBLR>1 Million).

DNA kinship analysis of unidentified human remains that led to a murder investigation.

DNA-based analysis is an important tool for the identification of unidentified human remains (UHR). Comparing DNA of UHR with DNA profiles of missing persons' relatives may provide genetic data leading to identification. In most cases, DNA profiles of missing persons are not available and therefore, the identification process must be based on kinship analysis. UHR were found buried in Southern Israel in 2016. Kinship analysis indicated potential family links to members of a Bedouin family. The Joint Pedigree Likelihood Ratio (JPLR) was lower than the familial database threshold and the match could easily have been missed due to its low value (245). A few factors led to expanded analysis, and the addition of more family members (from the forensic database), as well as the exclusion of half-sister, radically increased the JPLR value (4.94 × 1012). This case demonstrates the crucial importance of examining the metadata and analyzing the STRs regarding family links in cases of low JPLR, as well as the need to re-calculate the JPLR individually for each family member in the pedigree versus the UHR, and to increase the number of family members uploaded into the pedigree.

The rate of recurrent BRCA1, BRCA2, and TP53 mutations in the general population, and unselected ovarian cancer cases, in Belo Horizonte, Brazil.

In Brazil, several recurring mutations in BRCA1 and BRCA2 and a TP53 mutation (R337H) have been reported in high risk breast cancer cases. We hypothesized that these recurring mutations may also be detected in the general population and ovarian cancer cases in the state of Minas Gerais. To test this notion, participants were recruited from the outpatient and the Gynecological clinic in the UFMG Medical Center in Belo Horizonte, Minas Gerais, Brazil. BRCA1 (c.68_69delAG, c.5266dupC, c.181T>G, c.4034delA, c.5123C>A), BRCA2 (c.5946delT, c.8537_8538delAG, 4936_4939delGAAA), the c.156_157insAlu* BRCA2 and the c.1010G>A *TP53 mutation were genotyped using validated techniques. Overall, 513 cancer free participants (273 men) (mean age 47.7 ± 15.1 years) and 103 ovarian cancer cases (mean age at diagnosis 58.7 ± 9.6 years) were studied. None of the participants were found to carry any of the genotyped mutations. We conclude that the recurring mutations in BRCA1, BRCA2 and TP53 cannot be detected in the general population or consecutive ovarian cancer cases in this geographical region in Brazil.

The pre-GAP-related domain of neurofibromin regulates cell migration through the LIM kinase/cofilin pathway.

Neurofibromin contains several domains, most notably a GAP-related domain (GRD), that down-regulates Ras pathways. The functions of the non-GRD neurofibromin domains are largely known. Here we show that the pre-GRD region of neurofibromin alters the expression of genes involved in cell adhesion and migration and acts as a negative regulator of the Rac1/Pak1/LIMK1/cofilin pathway. Thus, neurofibromin-deficient glioblastoma and mouse fibroblasts are enriched in Rac1-GTP, p-Pak1, p-LIMK1 and p-cofilin, with all proteins exhibiting decreased expression upon expression of NF1(1-1163) polypeptide. Concomitantly, actin stress fibers and focal adhesion were disassembled and cell migration was halted. These effects were independent of the Ras signaling pathways. It seems that NF1(1-1163), through negative regulation of Rac-1, shifts the balance from a state of inactive phospho-cofilin to active unphosphorylated cofilin, resulting in severing of F-actin. Impairment of these cellular functions of neurofibromin provides novel insights into the invasiveness/progression of NF1-associated tumors.