The KRAS-Variant and miRNA Expression in RTOG Endometrial Cancer Clinical Trials 9708 and 9905

Objective

To explore the association of a functional germline variant in the 3′-UTR of KRAS with endometrial cancer risk, as well as the association of microRNA (miRNA) signatures and the KRAS-variant with clinical characteristics and survival outcomes in two prospective RTOG endometrial cancer trials.

Methods/Materials

The association of the KRAS-variant with endometrial cancer risk was evaluated by case-control analysis of 467 women with type 1 or 2 endometrial cancer and 582 age-matched controls. miRNA and DNA were isolated for expression profiling and genotyping from tumor specimens of 46 women with type 1 endometrial cancer enrolled in RTOG trials 9708 and 9905. miRNA expression levels and KRAS-variant genotype were correlated with patient and tumor characteristics, and survival outcomes were evaluated by variant allele type.

Results

The KRAS-variant was not significantly associated with overall endometrial cancer risk (14% controls and 17% type 1 cancers), although was enriched in type 2 endometrial cancers (24%, p = 0.2). In the combined analysis of RTOG 9708/9905, miRNA expression differed by age, presence of lymphovascular invasion and KRAS-variant status. Overall survival rates at 3 years for patients with the variant and wild-type alleles were 100% and 77% (HR 0.3, p = 0.24), respectively, favoring the variant.

Conclusions

The KRAS-variant may be a genetic marker of risk for type 2 endometrial cancers. In addition, tumor miRNA expression appears to be associated with patient age, lymphovascular invasion and the KRAS-variant, supporting the hypothesis that altered tumor biology can be measured by miRNA expression, and that the KRAS-variant likely impacts endometrial tumor biology.     

Divergence of water balance mechanisms in two sibling species (Drosophila simulans and D. melanogaster): effects of growth temperatures

Drosophila simulans is more abundant under colder and drier montane habitats in the western Himalayas as compared to its sibling D. melanogaster but the mechanistic bases of such climatic adaptations are largely unknown. Previous studies have described D. simulans as a desiccation sensitive species which is inconsistent with its occurrence in temperate regions. We tested the hypothesis whether developmental plasticity of cuticular traits confers adaptive changes in water balance-related traits in the sibling species D. simulans and D. melanogaster. Our results are interesting in several respects. First, D. simulans grown at 15 °C possesses a high level of desiccation resistance in larvae (~39 h) and in adults (~86 h) whereas the corresponding values are quite low at 25 °C (larvae ~7 h; adults ~13 h). Interestingly, cuticular lipid mass was threefold higher in D. simulans grown at 15 °C as compared with 25 °C while there was no change in cuticular lipid mass in D. melanogaster. Second, developmental plasticity of body melanisation was evident in both species. Drosophila simulans showed higher melanisation at 15 °C as compared with D. melanogaster while the reverse trend was observed at 25 °C. Third, changes in water balance-related traits (bulk water, hemolymph and dehydration tolerance) showed superiority of D. simulans at 15 °C but of D. melanogaster at 25 °C growth temperature. Rate of carbohydrate utilization under desiccation stress did not differ at 15 °C in both the species. Fourth, effects of developmental plasticity on cuticular traits correspond with changes in the cuticular water loss i.e. water loss rates were higher at 25 °C as compared with 15 °C. Thus, D. simulans grown under cooler temperature was more desiccation tolerant than D. melanogaster. Finally, desiccation acclimation capacity of larvae and adults is higher for D. simulans reared at 15 °C but quite low at 25 °C. Thus, D. simulans and D. melanogaster have evolved different strategies of water conservation consistent with their adaptations to dry and wet habitats in the western Himalayas. Our results suggest that D. simulans from lowland localities seems vulnerable due to limited acclimation potential in the context of global climatic change in the western Himalayas. Finally, this is the first report on higher desiccation resistance of D. simulans due to developmental plasticity of both the cuticular traits (body melanisation and epicuticular lipid mass) when grown at 15 °C, which is consistent with its abundance in temperate regions.     

Climatic adaptations of body melanisation in Drosophila melanogaster from Western Himalayas

We investigated population divergence in body melanisation in wild samples of Drosophila melanogaster across an elevational gradient (512-2202 m) in the Western Himalayas. Wild populations are characterized by higher phenotypic variability as compared with laboratory populations. Significant differences in elevational slope values for three posterior abdominal segments (fifth, sixth and seventh) in wild versus laboratory populations suggest plastic effects. However, elevational slope values do not differ for the three anterior abdominal segments (second, third and fourth). Thus, elevational changes in melanisation include genetic as well as plastic effects. Fitness consequences of within population variability were analyzed on the basis of assorted darker and lighter flies from two highlands as well as from two lowland localities. There is lack of correlation of melanisation with body size as well as ovariole number in assorted darker and lighter flies. For each population, darker flies showed higher desiccation resistance, lower rate of water loss, longer copulation duration and greater fecundity as compared with lighter flies. Phenotypic variations in body melanisation can be interpreted in relation with seasonal changes in temperature as well as humidity (Tcv and RHcv) of the sites of origin of populations. Thus, elevational changes in body melanisation may represent genetic response to selection pressures imposed by colder and drier climatic conditions in the Western Himalayas.     

p53 aberrations in low grade endometrioid carcinoma of the endometrium with nodal metastases: possible insights on pathogenesis discerned from immunohistochemistry

Background

TP53 mutations are rarely identified in low grade endometrioid carcinoma of the endometrium, and their pathogenic significance in such tumors is evidenced by the fact that TP53 aberrations have been associated with reduced recurrence-free survival in this subset of tumors. However, TP53 aberrations may not always represent a driving molecular event in a given endometrial cancer with a mutation. In this case study, the immunophenotype of a distinctive low grade endometrioid adenocarcinoma with an unusual pattern of lymph node metastases is used to explore the possible roles for underlying TP53-related molecular events in its pathogenesis.

Case presentation

A low grade endometrioid carcinoma, 9 cm in greatest dimension, with 35% invasion of the myometrial wall thickness, focal lymphovascular invasion, and metastases to 2 of 16 pelvic lymph nodes, was diagnosed in a 52-year-old woman. The endometrial tumor showed a p53-mutation (aberrant)-type immunohistochemical pattern in 40% of the tumor, but the rest of the tumor, as well as the foci of myometrial and lymphovascular invasion, were p53-wild type. Both lymph nodes with metastatic disease showed a distinct biphasic pattern, comprised of both p53-wild type and p53-aberrant areas in tumoral foci that were spatially apposed but not intermixed. Most p53-aberrant areas (at both the lymph nodes and the endometrium) showed a higher mitotic index and increased atypia as compared to the p53-wild type areas; both showed squamous differentiation. The p53-aberrant areas at both locations were also p16-diffusely positive, vimentin-positive, and estrogen/progesterone receptor-positive, whereas the p53-wild type areas showed an identical immunophenotype with the exception of being p16-mosaic positive. All components of the tumor at both the primary and metastatic sites showed loss of MSH2 and MSH6 and retained MLH/PMS2 expression.

Conclusions

The presence of p53-mutant and wild-type areas in multiple lymph nodes, coupled with the absence of a p53-aberrant immunophenotype in the myometrium-invasive or lymphovascular-invasive portions of the tumor, argues against the possibility that the TP53 mutation in this tumor is a driving event in its pathogenesis, at least regarding the metastatic process. This case illustrates how routine immunohistochemistry can provide important insights into underlying molecular events in cancers, exemplifies an uncommon co-existence of DNA mismatch repair protein deficiency and p53-aberrant immunophenotype in low-grade endometrioid carcinoma, illustrates morphologic differences between p53-aberrant and p53-wild type areas within in the same tumor, and is an exemplar of the emerging theory that lymph node metastases of endometrial cancer may be comprised of different subclones of the primary tumor.

     

Geographical Clines for Quantitative Traits in Natural Populations of a Tropical Drosophilid: Zaprionus Indianus

We analyzed natural populations of Zaprionus indianusin 10 Indian localities along a south-north transect (latitude: 10–31°3 N). Size traits (body weight, wing length and thorax length) as well as a reproductive trait (ovariole number) followed a pattern of clinal variation, that is, trait value increased with latitude. Wing/thorax ratio, which is inversely related to wing loading, also had a positive, but non-significant correlation with latitude. By contrast, bristle numbers (sternopleural and abdominal) exhibited a non-significant but negative correlation with latitude. Sex dimorphism, estimated as the female/male ratio, was very low in Z. indianus, contrasting with results already published in other species. Genetic variations among populations were also analyzed according to other geographic parameters (altitude and longitude) and to climatic conditions from each locality. A significant effect of altitude was found for size traits. For abdominal bristles, a multiple regression technique evidenced a significant effect of both latitude and altitude, but in opposite directions. Genetic variations were also correlated to climate, and mainly with average year temperature. Taking seasonal variations into account failed however to improve the predictability of morphometrical variations. The geographic differentiation of Z.indianusfor quantitative traits suggests adaptive response to local conditions, especially to temperature, but also reveals a complex situation according to traits investigated and to environmental parameters, which does not match results on other drosophilid species.     

Use of implantable cardioverter defibrillators after out-of-hospital cardiac arrest: a prospective follow-up study

Background

Survivors of out-of-hospital cardiac arrest are at high risk of recurrent arrests, many of which could be prevented with implantable cardioverter defibrillators (ICDs). We sought to determine the ICD insertion rate among survivors of out-of-hospital cardiac arrest and to determine factors associated with ICD implantation.

Methods

The Ontario Prehospital Advanced Life Support (OPALS) study is a prospective, multiphase, before–after study assessing the effectiveness of prehospital interventions for people experiencing cardiac arrest, trauma or respiratory arrest in 19 Ontario communities. We linked OPALS data describing survivors of cardiac arrest with data from all defibrillator implantation centres in Ontario.

Results

From January 1997 to April 2002, 454 patients in the OPALS study survived to hospital discharge after experiencing an out-of-hospital cardiac arrest. The mean age was 65 (standard deviation 14) years, 122 (26.9%) were women, 398 (87.7%) had a witnessed arrest, 372 (81.9%) had an initial rhythm of ventricular tachycardia or ventricular fibrillation (VT/VF), and 76 (16.7%) had asystole or another arrhythmia. The median cerebral performance category at discharge (range 1–5, 1 = normal) was 1. Only 58 (12.8%) of the 454 patients received an ICD. Patients with an initial rhythm of VT/VF were more likely than those with an initial rhythm of asystole or another rhythm to undergo device insertion (adjusted odds ratio [OR] 9.63, 95% confidence interval [CI] 1.31–71.50). Similarly, patients with a normal cerebral performance score were more likely than those with abnormal scores to undergo ICD insertion (adjusted OR 12.52, 95% CI 1.74–92.12).

Interpretation

A minority of patients who survived cardiac arrest underwent ICD insertion. It is unclear whether this low usage rate reflects referral bias, selection bias by electrophysiologists, supply constraint or patient preference.People who survive out-of-hospital cardiac arrest have an increased risk of recurrent arrest of 18%–20% in the first year.^1,2 Three large randomized studies evaluated the use of implantable cardioverter defibrillators (ICDs) versus antiarrhythmic drugs in survivors of out-of-hospital cardiac arrest.^3,4,5 The largest of the 3 studies involved 1016 patients and found a 39% relative risk reduction in mortality in the ICD group.³ The 2 smaller studies both reported nonsignificant reductions in mortality in the ICD group.^4,5 Two recent meta-analyses showed that the use of ICDs was associated with significant and important increases in survival among cardiac arrest survivors: all-cause mortality was reduced by 23%–28% with their use for secondary prevention, and the rate of sudden cardiac death was reduced by 50% in both meta-analyses.^6,7Guidelines from several national and international societies recommend insertion of ICDs in all survivors of cardiac arrest without a reversible cause.^8,9 Despite advances in ICD insertion and technology, studies to date suggest that the utilization rate is low, at least in some settings.^10,11 Several factors, including patient preference, physician referral, availability and cost, may contribute to the underutilization of ICDs.The Ontario Prehospital Advanced Life Support Study (OPALS)^12,13 is a multiphase before–after study designed to systematically evaluate the effectiveness of various prehospital interventions for people experiencing cardiac arrest, trauma or respiratory arrest. As an extension of the OPALS study, we sought to determine the rate of ICD insertion among survivors of cardiac arrest, as well as the factors associated with ICD implantation.