Pyruvate kinase is a dosage-dependent regulator of cellular amino acid homeostasis

The glycolytic enzyme pyruvate kinase (PK) is required for cancer development, and has been implicated in the metabolic transition from oxidative to fermentative metabolism, the Warburg effect. However, the global metabolic response that follows changes in PK activity is not yet fully understood. Using shotgun proteomics, we identified 31 yeast proteins that were regulated in a PK-dependent manner. Selective reaction monitoring confirmed that their expression was dependent on PK isoform, level and activity. Most of the PK targets were amino acid metabolizing enzymes or factors of protein translation, indicating that PK plays a global regulatory role in biosynthethic amino acid metabolism. Indeed, we found strongly altered amino acid profiles when PK levels were changed. Low PK levels increased the cellular glutamine and glutamate concentrations, but decreased the levels of seven amino acids including serine and histidine. To test for evolutionary conservation of this PK function, we quantified orthologues of the identified PK targets in thyroid follicular adenoma, a tumor characterized by high PK levels and low respiratory activity. Aminopeptidase AAP-1 and serine hydroxymethyltransferase SHMT1 both showed PKM2- concentration dependence, and were upregulated in the tumor. Thus, PK expression levels and activity were important for maintaining cellular amino acid homeostasis. Mediating between energy production, ROS clearance and amino acid biosynthesis, PK thus plays a central regulatory role in the metabolism of proliferating cells.     

Characterizing mammographic images by using generic texture features

Introduction

Although mammographic density is an established risk factor for breast cancer, its use is limited in clinical practice because of a lack of automated and standardized measurement methods. The aims of this study were to evaluate a variety of automated texture features in mammograms as risk factors for breast cancer and to compare them with the percentage mammographic density (PMD) by using a case-control study design.

Methods

A case-control study including 864 cases and 418 controls was analyzed automatically. Four hundred seventy features were explored as possible risk factors for breast cancer. These included statistical features, moment-based features, spectral-energy features, and form-based features. An elaborate variable selection process using logistic regression analyses was performed to identify those features that were associated with case-control status. In addition, PMD was assessed and included in the regression model.

Results

Of the 470 image-analysis features explored, 46 remained in the final logistic regression model. An area under the curve of 0.79, with an odds ratio per standard deviation change of 2.88 (95% CI, 2.28 to 3.65), was obtained with validation data. Adding the PMD did not improve the final model.

Conclusions

Using texture features to predict the risk of breast cancer appears feasible. PMD did not show any additional value in this study. With regard to the features assessed, most of the analysis tools appeared to reflect mammographic density, although some features did not correlate with PMD. It remains to be investigated in larger case-control studies whether these features can contribute to increased prediction accuracy.     

Correlation of optical coherence tomography with clinical and histopathological findings in experimental autoimmune uveoretinitis

Optical coherence tomography (OCT) is becoming the state-of-the-art method for the non-invasive imaging of a variety of ocular diseases. The aim of this study was to assess the application of OCT for the in vivo monitoring and follow-up of pathological changes during experimental autoimmune uveoretinitis (EAU) in rats. Initially we established OCT imaging in healthy brown Norway rats and correlated it with retinal histology. Subsequently, we induced EAU and imaged animals by OCT throughout the pre-peak, peak, and post-peak phases of the disease. The sensitivity of OCT imaging was determined by comparison with clinical EAU and histopathology scores obtained ex vivo at several time points throughout the disease course. Our data demonstrate that OCT imaging of the healthy rat retina closely correlates with histological observations and allows the clear visualization of all retinal layers. After induction of EAU, the first pathological changes could be detected by OCT at day (d) 8 post-immunization (p.i.) which corresponded to the time point of clinical disease onset. An increase in retinal thickness (RT) was detected from d10 p.i. onwards which peaked at d16 p.i. and decreased again to near control levels by d20 p.i. We introduce a novel semi-quantitative OCT scoring which correlates with histopathological findings and complements the clinical scores. Therefore, we conclude that OCT is an easily accessible, non-invasive tool for detection and follow-up of histopathological changes during EAU in rats. Indeed, significant differences in RT between different stages of EAU suggest that this OCT parameter is a sensitive marker for distinguishing disease phases in vivo.     

Impaired reproductive development in sons of women occupationally exposed to pesticides during pregnancy

Objectives

The aim of this prospective study was to investigate whether occupational pesticide exposure during pregnancy causes adverse effects on the reproductive development in the male infants.

Design and measurements

Pregnant women employed in greenhouses in Denmark were consecutively recruited, and 113 mother–son pairs were included. The mothers were categorized as occupationally exposed (91 sons) or unexposed (22 sons) to pesticides during pregnancy. Testicular position and volume, penile length, and position of urethral opening were determined at 3 months of age using standardized techniques. Concentrations of reproductive hormones in serum from the boys were analyzed.

Results

The prevalence of cryptorchidism at 3 months of age was 6.2% [95% confidence interval (CI), 3.0–12.4]. This prevalence was considerably higher than among Danish boys born in the Copenhagen area (1.9%; 95% CI, 1.2–3.0) examined by the same procedure. Boys of pesticide-exposed mothers showed decreased penile length, testicular volume, serum concentrations of testosterone, and inhibin B. Serum concentrations of sex hormone-binding globulin, follicle-stimulating hormone, and the luteinizing hormone:testosterone ratio were increased compared with boys of nonexposed mothers. For individual parameters, only the decreased penile length was statistically significant (p = 0.04). However, all observed effects were in the anticipated direction, and a joint multivariate test showed that this finding had a p-value of 0.012.

Conclusions

Our findings suggest an adverse effect of maternal occupational pesticide exposure on reproductive development in the sons despite current greenhouse safeguards and special measures to protect pregnant women.     

Radiofrequency Ablation of Liver Metastases—Software-Assisted Evaluation of the Ablation Zone in MDCT: Tumor-Free Follow-Up Versus Local Recurrent Disease

The purpose of this study was to investigate differences in change of size and CT value between local recurrences and tumor-free areas after CT-guided radiofrequency ablation (RFA) of hepatic metastases during follow-up by means of dedicated software for automatic evaluation of hepatic lesions. Thirty-two patients with 54 liver metastases from breast or colorectal cancer underwent triphasic contrast-enhanced multidetector-row computed tomography (MDCT) to evaluate hepatic metastatic spread and localization before CT-guided RFA and for follow-up after intervention. Sixteen of these patients (65.1 ± 10.3 years) with 30 metastases stayed tumor-free (group 1), while the other group (n = 16 with 24 metastases; 62.0 ± 13.8 years) suffered from local recurrent disease (group 2). Applying an automated software tool (SyngoCT Oncology; Siemens Healthcare, Forchheim, Germany), size parameters (volume, RECIST, WHO) and attenuation were measured within the lesions before, 1 day after, and 28 days after RFA treatment. The natural logarithm (ln) of the quotient of the volume 1 day versus 28 days after RFA treament was computed: lnQ1//28/0_volume. Analogously, ln ratios of RECIST, WHO, and attenuation were computed and statistically evaluated by repeated-measures ANOVA. One lesion in group 2 was excluded from further evaluation due to automated missegmentation. Statistically significant differences between the two groups were observed with respect to initial volume, RECIST, and WHO (p < 0.05). Furthermore, ln ratios corresponding to volume, RECIST, and WHO differed significantly between the two groups. Attenuation evaluations showed no significant differences, but there was a trend toward attenuation assessment for the parameter lnQ28/0_attenuation (p = 0.0527), showing higher values for group 1 (–0.4 ± 0.3) compared to group 2 (–0.2 ± 0.2). In conclusion, hepatic metastases and their zone of coagulation necrosis after RFA differed significantly between tumor-free and local-recurrent ablation zones with respect to the corresponding size parameters. A new parameter (lnQ1//28/0_{volume/RECIST/WHO/attenuation}) was introduced, which appears to be of prognostic value at early follow-up CT.