Elevated level of pro-inflammatory eicosanoids and EPC dysfunction in diabetic patients with cardiac ischemia.

BACKGROUND: Circulating endothelial progenitor cells (EPCs) are recruited from the blood system to sites of ischemia and endothelial damage, where they contribute to the repair and development of blood vessels. Since numerous eicosanoids including leukotrienes (LTs) and hydroxyeicosatetraenoic acids (HETEs) have been shown to exert potent pro-inflammatory activities, we examined their levels in chronic diabetic patients with severe cardiac ischemia in conjunction with the level and function of EPCs. RESULTS: Lipidomic analysis revealed a diabetes-specific increase (p<0.05) in inflammatory and angiogenic eicosanoids including the 5-lipoxygenase-derived LTB (4.11±1.17 vs. 0.96±0.27 ng/ml), the lipoxygenase/CYP-derived 12-HETE (117.08±35.05 vs. 24.34±10.03 ng/ml), 12-HETrE (17.56±4.43 vs. 4.15±2.07 ng/ml), and the CYP-derived 20-HETE (0.32±0.04 vs. 0.06±0.05 ng/ml) the level of which correlated with BMI (p=0.0027). In contrast, levels of the CYP-derived EETs were not significantly (p=0.36) different between these two groups. EPC levels and their colony-forming units were lower (p<0.05) with a reduced viability in diabetic patients compared with non-diabetics. EPC function (colony-forming units (CFUs) and MTT assay) also negatively correlated with the circulating levels of HgA1C. CONCLUSION: This study demonstrates a close association between elevated levels of highly pro-inflammatory eicosonoids, diabetes and EPC dysfunction in patients with cardiac ischemia, indicating that chronic inflammation impact negatively on EPC function and angiogenic capacity in diabetes.

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.

Technical note: The Next Step - a semi-automatic coding and comparison system for forensic footwear impressions.

Footwear impression evidence is a key tool in criminal investigations, connecting suspects to the crime scene; in addition, it may provide valuable forensic intelligence linking different crime scenes in the absence of a suspect. This paper presents the development of a computer database and semi-automatic system for shoeprint comparison. The database is based on structured manual coding of elements by CSI (Crime Scene Investigators). The computer algorithm then compares the pattern information of the query footwear impression to the entire database, returning a list of possible matches ranked by pattern similarity and crime scene geographic proximity. Initial results using a database of 284 footwear impressions from real crime scenes revealed that a "match" (i.e., the same shoe model) was found for 30% of the impressions; in most cases, the "match" ranked within the top five places of the "hit" list generated by the algorithm. Our results confirm that this semi-automatic footwear comparison system is simple, cost-effective and efficient, providing great potential for linking crime scenes.

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.

Investigation of Different Ways in Which the CODIS 7.0 May be Used in Mass Disaster Identification.

DNA analysis is a key method for the identification of human remains in mass disasters. Reference samples from relatives may be used to identify missing persons by kinship analysis. Different methods of applying the CODIS in disaster victim identification (DVI) were investigated. Two searches were evaluated: (i) relating family relatives to a pedigree tree (FPT) and (ii) relating unidentified human remains to a pedigree tree (UPT). A joint pedigree likelihood ratio (JPLR) and rank were calculated for each search. Both searches were similar in average JPLR and rank. In exceptional cases, namely the existence of a mutation different from the CODIS model, a nonbiological father, a mistake in STR, or incorrect profile association, the UPT search returned one true rank, whereas the FPT search returned no results. This paper suggests a novel strategy to overcome these limitations and increase efficiency in conducting identification of mass disaster victims.

Heme oxygenase-1 induction improves cardiac function following myocardial ischemia by reducing oxidative stress.

BACKGROUND: Oxidative stress plays a key role in exacerbating diabetes and cardiovascular disease. Heme oxygenase-1 (HO-1), a stress response protein, is cytoprotective, but its role in post myocardial infarction (MI) and diabetes is not fully characterized. We aimed to investigate the protection and the mechanisms of HO-1 induction in cardiomyocytes subjected to hypoxia and in diabetic mice subjected to LAD ligation. METHODS: In vitro: cultured cardiomyocytes were treated with cobalt-protoporphyrin (CoPP) and tin protoporphyrin (SnPP) prior to hypoxic stress. In vivo: CoPP treated streptozotocin-induced diabetic mice were subjected to LAD ligation for 2/24 h. Cardiac function, histology, biochemical damage markers and signaling pathways were measured. RESULTS: HO-1 induction lowered release of lactate dehydrogenase (LDH) and creatine phospho kinase (CK), decreased propidium iodide staining, improved cell morphology and preserved mitochondrial membrane potential in cardiomyocytes. In diabetic mice, Fractional Shortening (FS) was lower than non-diabetic mice (35±1%vs.41±2, respectively p<0.05). CoPP-treated diabetic animals improved cardiac function (43±2% p<0.01), reduced CK, Troponin T levels and infarct size compared to non-treated diabetic mice (P<0.01, P<0.001, P<0.01 respectively). CoPP-enhanced HO-1 protein levels and reduced oxidative stress in diabetic animals, as indicated by the decrease in superoxide levels in cardiac tissues and plasma TNFα levels (p<0.05). The increased levels of HO-1 by CoPP treatment after LAD ligation led to a shift of the Bcl-2/bax ratio towards the antiapoptotic process (p<0.05). CoPP significantly increased the expression levels of pAKT and pGSK3ß (p<0.05) in cardiomyocytes and in diabetic mice with MI. SnPP abolished CoPP's cardioprotective effects. CONCLUSIONS: HO-1 induction plays a role in cardioprotection against hypoxic damage in cardiomyocytes and in reducing post ischemic cardiac damage in the diabetic heart as proved by the increased levels of pAKT with a concomitant inhibition of pGSK3ß leading to preserved mitochondrial membrane potential.

Endothelial progenitor cell function inversely correlates with long-term glucose control in diabetic patients: association with the attenuation of the heme oxygenase-adiponectin axis.

BACKGROUND: Endothelial progenitor cells (EPCs) are attenuated, both in number and functionality, in animal models of chronic cardiovascular and metabolic disorders. This effect has subsequently been linked to the aggravation of long-term morbidity and mortality associated with such disorders. The objective was to examine EPC number and survival in chronic diabetic vs nondiabetic patients in conjunction with the examination of their redox, inflammatory, and antioxidant defense system (Nrf2 genes) status in serum and visceral fat. METHODS: Visceral adipose tissue from diabetic and nondiabetic patients undergoing coronary artery bypass graft surgery was analyzed for Nrf2-dependent genes. Oxidative stress was evaluated using thiobarbituric acid-reactive substance assay (TBARS). Peripheral blood, collected 1 day prior to surgery, was evaluated for inflammatory cytokines and EPCs. RESULTS: When compared with controls (P < 0.05), results of the thiobarbituric acid-reactive substance assay were higher in diabetic patients. Although Nrf2-dependent antioxidant proteins (thioredoxin-1 [Trx-1], nicotinamide adenine dinucleotide phosphate [NAD(P)H] quinone oxidoreductase [NQO1], glutathione S-transferase [GST]) were upregulated, heme oxygenase (HO-1) and adiponectin protein expression were lower in the diabetic group (P < 0.05). Serum levels of bilirubin were lower (P < 0.005) while the levels of inflammatory cytokines were higher in diabetic patients (P < 0.05). EPC levels and their colony forming units were significantly lower (P < 0.05) with reduced viability in diabetic patients as compared with nondiabetic patients. CONCLUSIONS: These results demonstrate for the first time that in diabetic patients, there is an inadequate heme oxygenase-adiponectin axis response, which could compromise the compensatory antioxidant and anti-inflammatory effects consequently contributing toward EPC dysfunction in these patients.