Effect of Raman exposure time on the quantitative and discriminant analyses of carotenoid concentrations in intact tomatoes.

The significant worldwide expansion of the health food market, which includes functional fruits and vegetables, requires a simple and rapid analytical method for the on-site analysis of functional components, such as carotenoids, in fruits and vegetables, and Raman spectroscopy is a powerful candidate. Herein, we clarified the effects of Raman exposure time on quantitative and discriminant analysis accuracies. Raman spectra of intact tomatoes with various carotenoid concentrations were acquired and used to develop partial least squares regression (PLSR) and partial least squares discriminant analysis (PLS-DA) models. The accuracy of the PLSR model was superior (R2 = 0.87) when Raman spectra were acquired 10 s, but decreased with decreasing exposure time (R2 = 0.69; 0.7 s). The accuracy of the PLS-DA model was unaffected by exposure time (hit rate: 90%). We conclude that Raman spectroscopy combined with PLS-DA is useful for the on-site analysis of carotenoids in fruits and vegetables.

Useful tissues in cabbage head for freshness evaluation with visible and near infrared spectroscopy.

To determine which cabbage head tissues are useful for evaluating freshness using spectroscopic technology, we stored wrapped and unwrapped cabbage heads for up to 30 d, and measured visible and near infrared spectra (420-2500 nm) of the 1st-10th leaf layers and cores. We found that spectral changes in leaves were affected both by leaf layer and storage conditions, while continuous spectral changes were observed in the cores regardless of storage condition. These spectral changes in the leaves and cores were consistent with color images and water content. While we developed good models for estimating the storage days from the 1st and 2nd leaf layers and the cores of unwrapped cabbages, only core spectra provided a high correlation with storage days in wrapped cabbages. These data demonstrated that the cabbage core is sensitive to storage duration and its spectra are useful for evaluating freshness decline regardless of storage condition.

Membrane topology of murine glycerol-3-phosphate acyltransferase 2.

Glycerol-3-phosphate acyltransferase (GPAT) is a rate-limiting enzyme in mammalian triacylglycerol biosynthesis. GPAT is a target for the treatment of metabolic disorders associated with high lipid accumulation. Although the molecular basis for GPAT1 activation has been investigated extensively, the activation of other isoforms, such as GPAT2, is less well understood. Here the membrane topology of the GPAT2 protein was examined using an epitope-tag-based method. Exogenously expressed GPAT2 protein was present in the membrane fraction of transformed HEK293 cells even in the presence of Na(2)CO(3) (100 mM), indicating that GPAT2 is a membrane-bound protein. Trypsin treatment of the membrane fraction degraded the N-terminal (FLAG) and C-terminal (myc-epitope) protein tags of the GPAT2 protein. Bioinformatic analysis of the GPAT2 protein sequence indicated four hydrophobic sequences as potential membrane-spanning regions (TM1-TM4). Immunoblotting of the myc-epitope tag, which was inserted between each TM region of the GPAT2 protein, showed that the amino acid sequence between TM3 and TM4 was protected from trypsin digestion. These results suggest that the GPAT2 protein has two transmembrane segments and that the N-terminal and C-terminal regions of this protein face the cytoplasm. These results also suggest that the enzymatically active motifs I-III of the GPAT2 protein face the cytosol, while motif IV is within the membrane. It is expected that the use of this topological model of GPAT2 will be essential in efforts to elucidate the molecular mechanisms of GPAT2 activity in mammalian cells.

Alternative splicing produces a constitutively active form of human SREBP-1.

We identified a novel alternative splicing event that constitutively produces a truncated active form of human sterol regulatory element-binding protein 1 (SREBP-1). A cDNA of this splicing variant (named SREBP-1Delta) contains a translational stop codon-encoding exon sequence between exons 7 and 8. It produces SREBP-1aDelta (470 a.a.) and SREBP-1cDelta (446 a.a.) proteins that lack transmembrane and C-terminal regulatory sequences necessary for localization of SREBP-1 to the endoplasmic reticulum. A luciferase reporter assay showed that SREBP-1aDelta and SREBP-1cDelta transactivated lipogenic gene promoters to the same extent as that induced by N-terminal active fragments of SREBP-1a and SREBP-1c, respectively. SREBP-1Delta mRNA is expressed in human cell lines as well as adipose and liver tissues. Expression levels ranged from 5% to 16% of total SREBP-1 expression. The ratio of SREBP-1Delta expression to total SREBP-1 expression in HepG2 cells was not affected by either insulin or high glucose treatment.

Relationship between the appearance of preantral follicles in the fetal ovary of Antarctic minke whales (Balaenoptera bonaerensis) and hormone concentrations in the fetal heart, umbilical cord and maternal blood.

The present study aimed to determine the relationship among changes in the number of preantral follicles and concentrations of follicle-stimulating hormone (FSH), luteinizing hormone (LH), progesterone (P4), androstenedione (A) and estradiol-17beta (E2) in the fetal heart, umbilical cord and maternal blood. Primordial follicles had already appeared in a 20 cm fetus and primary follicles were observed in a 50 cm fetus. In a 70 cm fetus, the number of primordial and primary follicles increased rapidly and secondary follicles were present. The concentrations of LH and FSH did not change between 20 cm and 160 cm in fetal length. When the fetal length became > 70 cm, serum levels in the fetus, umbilical cord and mothers, and E2 levels in umbilical cord increased synchronously (p < 0.05). These results showed increases in the number of preantral follicles in the Antarctic minke whale fetal ovary along with fetal growth during the early gestation period. These findings suggest that the change in preantral follicles was associated with changes in the concentration of steroids in early gestation periods. The changes in steroid concentrations in the fetal and umbilical cord blood and the increased number of preantral follicles were coincident at around 70 cm in fetal length, whereas the growth and differentiation of primordial and primary follicles appeared to be independent of FSH and LH.