Bioavailability and Biological Effects of 2- O-ß-d-Glucopyranosyl-carboxyatractyligenin from Green Coffee in Caenorhabditis elegans.

Targeted analysis of Coffea arabica and Coffea canephora green coffees (total sample size n = 57) confirmed 2- O-ß-d-glucopyranosyl-carboxyatractyligenin (6) as the quantitatively dominating carboxyatractyligenin derivative. Its abundance in Arabicas (2425 ± 549 nmol/g, n = 48) exceeded that in Robustas (34 ± 12 nmol/g, n = 9) roughly by a factor of 70. Coffee processing involving heat (e.g., steam treatment and decaffeination) reduced concentrations of 6 and increased those of the decarboxylated derivative. The bioavailability of compound 6 in Caenorhabditis elegans was demonstrated by ultraperformance liquid chromatography-tandem mass spectrometry analysis of extracts prepared from nematode cultures incubated in a liquid medium containing 6. A toxicity assay performed to assess the impact of 6 in vivo showed a 20-fold higher median lethal dose (LD50 = 11.7 ± 1.2 mM) concentration compared to that of the known phytotoxic adenine-nucleotide transporters inhibitor carboxyatractyloside (2, LD50 = 0.61 ± 0.05 mM), whereas 1 mM 6 and 0.1 mM 2 were sufficient to decrease the survival of wild type C. elegans, already 10-20-fold lower doses reduced reproduction. Because the insulin/insulin-like growth factors signaling cascade (IIS) is a key regulator of life span and stress resistance, the impact of compound 6 on the survival of long-living daf-2 C. elegans was tested. As the susceptibility of these nematodes to 6 was as high as that in wild type, an impact on central metabolic processes independent of IIS was suggested. Analysis of the in vivo adenosine triphosphate (ATP) content of adult C. elegans revealed no changes after 1 and 24 h, but a 50% reduction after treatment with 1 mM 6 during the entire postembryonic development. These data speak for a developmental-stage-dependent modulation of the ATP pool by 6.

The Reproduction Rate of Peptide Transporter PEPT-1 Deficient C. elegans Is Dependent on Dietary Glutamate Supply.

Intestinal absorption of dietary amino acids is mediated via two routes. Free amino acids released by hydrolysis of dietary proteins are taken up by a multitude of amino acid transporters while di- and tripeptides released are taken up by the peptide transporter PEPT-1. Loss of PEPT-1 impairs growth, post-embryonic development and reproduction in Caenorhabditis elegans, and supplementation with a mixture of all L-amino acids only partially rescues fertility. In the present study, we demonstrate that dietary L-glutamate is the responsible amino acid that can increase fertility in hermaphrodite pept-1 worms. This effect was associated with a significantly higher uptake of glutamate/aspartate in pept-1 than in wildtype C. elegans. Furthermore, we found that the intestinal transporter proteins SNF-5 of the solute carrier SLC6 family of nutrient amino acid transporters (NAT) and AAT-6 of the SLC7 family as the light subunit of a heteromeric amino acid transporter (HAT) play a key role in glutamate homeostasis in pept-1 C. elegans. Genes encoding these transporters are highly expressed and upon silencing a 95% reduced fertility (snf-5) and sterility (aat-6) was observed. A subsequent L-glutamate supplementation failed to rescue these phenotypes. Dietary glutamate supplementation did neither influence the feeding frequency, nor did it improve mating efficiency of pept-1 males. Most strikingly, pept-1 were more prone to habituation to repeated gentle touch stimuli than wildtype C. elegans, and dietary glutamate supply was sufficient to alter this behavioral output by restoring the mechanosensory response to wildtype levels. Taken together, our data demonstrate a key role of L-glutamate in amino acid homeostasis in C. elegans lacking the peptide transporter in the intestine and demonstrate its distinct role in reproduction and for neural circuits mediating touch sensitivity.

Relation between maternal angiogenic factors and utero-placental resistance in normal first- and second-trimester pregnancies.

OBJECTIVE: Soluble endoglin (sEng) is a novel antiangiogenic protein and elevated sEng concentrations in maternal circulation are closely related to preeclampsia and HELLP syndrome. As the perfusion of the uterine arteries as well as the dynamics of angiogenic factors between first and second trimester have prognostic value regarding pregnancy outcome, it was the aim of this study to investigate the relation between maternal angiogenic factors and uterine Doppler parameters. STUDY DESIGN: The longitudinal study includes 50 normal pregnancies. Pulsatility index (PI) of the uterine arteries was detected by Doppler ultrasound in first and second trimester. In parallel, maternal sEng and soluble fms-like tyrosine kinase 1 (sFlt1) concentration was measured using ELISA. RESULTS: In the first trimester, the sEng concentrations were 4.92 ± 1.36 ng/mL and the uterine PI was 1.14 ± 0.28. In the second trimester, the maternal sEng concentration decreased significantly to 3.99 ± 0.63 ng/mL (p < 0.05) which was associated by a decrease of the uterine PI to 0.78 ± 0.15 (p < 0.001). Soluble fms-like tyrosine kinase 1 concentrations did not differ significantly between first and second trimester (423 ± 333 vs. 444 ± 291 pg/mL). There was a significant negative correlation between sEng and uterine resistance in the second trimester (r = -0.416; p < 0.001). CONCLUSIONS: In normal pregnancy, parallel to the fall of utero-placental resistance, there is a physiological decline of the maternal sEng concentration between first and second trimester. In second trimester, there is a negative correlation between sEng and uterine Doppler parameters.

The value of positron emission tomography in the follow-up for breast cancer.

BACKGROUND: The aim of this study was to investigate the diagnostic value of FDG PET in the follow-up for breast cancer in disease-free patients and patients suspected of having recurrent or metastatic disease. As a single imaging tool, PET can be compared with the conventional diagnostic means used for different examination sites. PATIENTS AND METHODS: A total of 73 PET studies were carried out on 57 patients who had been diagnosed as having breast cancer. Sixteen patients had two PET scans. Thirty-eight scans (52.%) were performed in a follow-up setting. Thirty-five PET scans were performed in patients suspected of having recurrent disease or elevated tumor marker. Depending on the region of suspicion, conventional imaging included computed tomography, magnetic resonance imaging, chest X-ray, ultrasound and X-ray. All the patients in our study were followed-up for a period of 12 months. RESULTS: PET correctly identified metastatic or recurrent disease in 25 out 27 cases of clinical suspicion. In patients examined because of elevated tumour marker CA 15-3, PET was able to detect recurrence or metastatic disease in six of the eight patients. The absence of disease was correctly diagnosed by PET in 35 out of 38 scans in 24 patients in the follow-up for breast cancer. The overall sensitivity and specificity for PET was 80.6% and 97.6%, respectively. On the basis of a yearly rate of disease progression of 5-8%, the mean positive predictive value was 74.5% and the mean negative predictive value 98.3%. CONCLUSION: PET has been shown to have impact on the staging and management of recurrent or metastatic breast cancer in cases of suspicion and in a follow-up setting. The current oncological situation can be clarified with a single basic imaging modality.