Polymorphism of 9p21.3 Locus Is Associated with 5-Year Survival in High-Risk Patients with Myocardial Infarction

Objective

The rs10757278, rs1333049 and rs4977574 are single nucleotide polymorphisms (SNPs) of chromosome 9p21 locus associated with a prevalence of acute coronary syndromes (ACS). Reports concerning their association with long-term outcome after an ACS are equivocal. The aim of our study was to investigate the association of the 9p21.3 locus with 5-year overall mortality in patients with ST-elevation myocardial infarction (STEMI).

Materials and methods

We performed a retrospective analysis of data collected prospectively in 2 independent registries of consecutive patients with STEMI (derivation and validation group). Genotyping was performed with the TaqMan method. The analyzed end-point was total mortality.

Results

The derivation group comprised 589 patients: 25.3% female (n = 149), mean age 62.4±12.0 years, total 5-year mortality 16.6% (n = 98). When all the study group was analyzed, no significant differences in mortality were found between the genotypes. However, in high-risk patients (GRACE risk score ≥155 points, n = 238), homozygotes associated with higher risk for ACS had significantly better 5-year survival compared to other genotypes. The hazard ratio associated with the high-risk genotype (a homozygote of high risk for ACS or a heterozygote) was: HR = 2.2 (1.15–4.2) for the rs10757278 polymorphism, HR = 2.7 (95% CI 1.3–5.4) for the rs4977574 one and HR = 2.3 (1.2–4.5) for the rs1333049 one (Cox proportional hazards model). Survival analysis in the validation group (n = 365) showed a clear trend towards better prognosis in GG homozygotes of the rs10757278 SNP, which confirms our initial results (p = 0.09, log-rank test).

Conclusions

The 9p21.3 locus is associated with 5-year mortality in high-risk patients with STEMI. The genotypes associated with higher risk for ACS show a protective effect in terms of further survival (instead of a deteriorating prognosis, as reported previously). This finding, due to the very high size of the effect, could potentially be applied to clinical practice, if appropriate methods are elaborated.     

Flow cytometric analysis of CD55 and CD59 expression on blood cells in paroxysmal nocturnal haemoglobinuria

PNH is a rare clonal disorder of hematopoietic stem cells, therefore all blood cells lineages are involved. The main feature is an increased sensitivity of erythrocytes to complement-mediated cell lysis due to deficiency of membrane-bound GPI (glycosylphosphatidylinositol)-anchored proteins which normally function as inhibitors of reactive hemolysis. In the present study, we performed flow cytometric analysis using monoclonal antibodies against CD55 and CD59 for the detection of PNH-type clone in the blood of 50 patients (28 females and 22 males, age range 7-67 yrs). In one patient only we found a large population (95%) of granulocytes with decreased expression of both CD55 and CD59 molecules (type I PNH) and in two others with partial loss of CD55 expression (type II PNH). The expression was determined chiefly on granulocytes which in the control group showed reliable and high expression of CD55 and CD59.     

Recycling of manure nutrients: use of algal biomass from dairy manure treatment as a slow release fertilizer

An alternative to land spreading of manure is to grow crops of algae on the N and P present in the manure and convert manure N and P into algal biomass. The objective of this study was to evaluate the fertilizer value of dried algal biomass that had been grown using anaerobically digested dairy manure. Results from a flask study using two soils amended with algal biomass showed that 3% of total algal nitrogen (N) was present as plant available N at day 0. Approximately 33% of algal N was converted to plant available N within 21 days at 25 degrees C in both soils. Levels of Mehlich-3 extractable phosphorus (P) in the two soils rose with increasing levels of algal amendment but were also influenced by existing soil P levels. Results from plant growth experiments showed that 20-day old cucumber and corn seedlings grown in algae-amended potting mix contained 15-20% of applied N, 46-60% of available N, and 38-60% of the applied P. Seedlings grown in algae-amended potting mixes were equivalent to those grown with comparable levels of fertilizer amended potting mixes with respect to seedling dry weight and nutrient content. These results suggest that dried algal biomass produced from treatment of anaerobically digested dairy manure can substitute for commercial fertilizers used for potting systems.     

Use Reference Bands to Accurately Estimate ISCN Band Levels 400, 550, and 850

In their 2002 Guidelines for chromosome analysis of peripheral blood, the American College of Medical Genetics states that "The 550-band stage should be the goal of all constitutional studies..." The College of American Pathologists requires that the average case be analyzed at the 400-band level of resolution for routine work, and that the 550-band level be achieved in appropriate blood samples. The challenge is how to identify the 400, 550, and 850-band levels confidently and consistently. In this study, our objectives were to develop simple and reliable criteria to estimate band level, and to evaluate our laboratory's performance with respect to those criteria. Using the ISCN(1995) ideogram as a reference, candidate bands were selected for the three band levels: 400, 550 and 850. A pilot and two follow-up studies were conducted and a set of candidate bands were validated against the Vancouver method of evaluating band level so that band level scores were similar using either method. The final set of reference bands were the presence of 9q32 and 20q13.2 for the 400-band level; 5q33.2 and 10q22.2 for the 550-band level; and 3p26.1, 18q22.3 and 20q13.32 for the 850-band level. Cell selection improved after each technologist was provided a composite image of chromosomes with reference bands highlighted. The band level criteria presented here involve no band counting, appear to be objective, can help to improve quality and consistency among technologists, and can ensure compliance with regulatory agencies.     

Using numerical approximation as an intermediate step in analytical derivations: some observations from biomechanics

We present four examples to illustrate the use of a type of numerical approximation as an intermediate step in analytical derivation of seemingly complicated biomechanical equations. The method involves examination of curve shapes to elucidate useful underlying trends, which may otherwise be overlooked through consideration of only the equations themselves. Two examples of the method's use are drawn from recently published results in the area of experimental methods in biomechanics of very soft tissues, and two others are taken from our current work on cartilage tissue mechanics. We think that such observations provide a useful means of circumventing complexity issues when deriving models for biomechanical analysis, and further that the method, while simple in concept, could be effective in a range of biomechanics applications.     

Cell-surface-associated tissue transglutaminase is a target of MMP-2 proteolysis

MT1-MMP, a prototypic member of a membrane-type metalloproteinase subfamily, is an invasion promoting protease and an activator of MMP-2. In addition, MT1-MMP proteolysis regulates the functionality of cell-surface adhesion/signaling receptors including tissue transglutaminase (tTG). tTG is known to serve as an adhesion coreceptor for beta1/beta3 integrins and as an enzyme that catalyzes the cross-linking of proteins and the conjugation of polyamines to proteins. Here, we report that MMP-2, functioning in concert with MT1-MMP, hydrolyzes cell-surface-associated tTG, thereby further promoting the effect initiated by the activator of MMP-2. tTG, in return, preferentially associates with the activation intermediate of MMP-2. This event decreases the rate of MMP-2 maturation and protects tTG against proteolysis by MMP-2. Our cell culture, in vitro experiments, and in silico modeling indicate that the catalytic domain of MMP-2 directly associates with the core enzymatic domain II of tTG (the K(d) = 380 nM). The follow-up cleavage of the domain II eliminates both the receptor and the enzymatic activity of tTG. Our data illuminate the coordinated interplay involving the MT1-MMP/MMP-2 protease tandem in the regulation of the cell receptors and explain the underlying biochemical mechanisms of the extensive tTG proteolysis that exists at the normal tissue/tumor boundary. Our findings also suggest that neoplasms, which express functionally active MT1-MMP and, therefore, activate soluble MMP-2, can contribute to the degradation of tTG expressed in neighboring host cells. The loss of adhesive and enzymatic activities of tTG at the interface between tumor and normal tissue will decrease cell-matrix interactions and inhibit matrix cross-linking, causing multiple pathological alterations in host cell adhesion and locomotion.     

Origin of the disjunct tetraploid Cardamine amporitana (Brassicaceae) assessed with nuclear and chloroplast DNA sequence data

Seventy-four nucleotide sequences from the ITS regions of nuclear ribosomal DNA and 76 from the trnL-trnF spacer of chloroplast DNA were used to address the origin of tetraploid Cardamine amporitana, the conspecifity of central Italian and northeastern Spanish populations, and the possible cause for such geographic disjunction. Because of the complex lineage relationships in Cardamine, the sampling included 22 taxa. In the results, both data sets are highly congruent in supporting a close relationship of C. amporitana to the widespread Eurasian C. amara. Low genetic variability in northeastern Spanish populations of C. amporitana suggests long-distance dispersal from central Italy. The interior position of the single northeastern Spanish haplotype in a statistical parsimony network of trnL-trnF haplotypes however does not support this scenario and invokes other plausible phylogeographic explanations. The disappearance of geographically intermediate populations and genetic impoverishment by migration and isolation, both probably associated with Quaternary climatic oscillations, appears as an alternative hypothesis to explain the phylogeographic pattern. A recent hybridization event is reported between C. amporitana and a diploid from the C. pratensis group in central Italy on the basis of additive polymorphisms in ITS for all the 22 distinguishing nucleotides.