Maternal undernutrition during early pregnancy inhibits postnatal growth of the tibia in the female offspring of rats by alteration of chondrogenesis.

Epidemiological research has suggested that birth weights are correlated with adult leg lengths. However, the relationship between prenatal undernutrition (UN) and postnatal leg growth remains controversial. We investigated the effects of UN during early pregnancy on postnatal hindlimb growth and determined whether early embryonic malnutrition affects the functions of postnatal chondrocytes in rats. Undernourished Wistar dams were fed 40% of the daily intake of rats in the control groups from gestational days 5.5-11.5, and femurs, tibias, and trunks or spinal columns were morphologically measured at birth and at 16 weeks of age in control and undernourished offspring of both sexes. We evaluated cell proliferation and differentiation of cultured chondrocytes derived from neonatal tibias of female offspring and determined chondrocyte-related gene expression levels in neonatal epiphysis and embryonic limb buds. Tibial lengths of undernourished female, but not male, offspring were longer at birth and shorter at 16 weeks of age (p < .05) compared with those of control rats. In chondrocyte culture studies, stimulating effects of IGF-1 on cell proliferation (p < .01) were significantly decreased and levels of type II collagen were lower in female undernourished offspring (p < .05). These phenomena were accompanied by decreased expression levels of Col2a1 and Igf1r and increased expression levels of Fgfr3 (p < .05), which might be attributable to the decreased expression of specificity protein 1 (p < .05), a key transactivator of Col2a1 and Igf1r. In conclusion, UN stress during early pregnancy reduces postnatal tibial growth in female offspring by altering the function of chondrocytes, likely reflecting altered expression of gene transactivators.

Stabilization technique for real-time high-resolution vascular ultrasound using frequency domain interferometry.

We have proposed an ultrasound imaging method based on frequency domain interferometry (FDI) with an adaptive beamforming technique to depict real-time high-resolution images of human carotid artery. Our previous study has investigated the performance of the proposed imaging method under an ideal condition with a high signal-to-noise ratio (SNR). In the present study, we propose a technique that has the potential to improve accuracy in estimating echo intensity using the FDI imaging method. We investigated the performance of the proposed technique in a simulation study that two flat interfaces were located at depths of 15.0 and 15.2 mm and white noise was added. Because the -6 dB bandwidth of the signal used in this simulation study is 2.6 MHz, the conventional B-mode imaging method failed to depict the two interfaces. Both the conventional and proposed FDI imaging methods succeeded to depict the two interfaces when the SNR ranged from 15 to 30 dB. However, the average error of the estimated echo intensity at the interfaces using the conventional FDI imaging method ranged from 7.2 to 10.5 dB. In contrast, that using the FDI imaging method with the proposed technique ranged from 2.0 to 2.2 dB. The present study demonstrates the potential of the FDI imaging method in depicting robust and high-range-resolution ultrasound images of arterial wall, indicating the possibility to improve the diagnosis of atherosclerosis in early stages.

Real-time high-resolution vascular ultrasound using frequency domain interferometry with the ROI-division process.

We have proposed a high-range-resolution ultrasound imaging method for human carotid artery using an adaptive beamforming technique based on frequency domain interferometry (FDI). The method assumes that the received signal consists of multiple echoes of targets and noise, where the waveform of each echo is similar to that of the reference signal. In this study, we examine the dependence of the echo waveform on the target depth, and investigate the proper measurement-range for the FDI imaging method using a reference signal. Furthermore, we propose a ROI-division process, where each sub-ROI has a proper measurement-range for the application of the FDI imaging method. Simulation and experimental results show the efficiency of the ROI-division process in improving the image quality of human carotid artery acquired using the FDI imaging method. We believe that the modified FDI imaging method with the ROI-division process has the potential to facilitate significant progress in the detection of vessel stenosis and in the assessment of cardiovascular disease risk.

Right hepatic artery traveling anteriorly to the common bile duct.

The topographic relationship between arteries and hepatobiliary ducts can be crucial during cholecystectomy. We observed the right hepatic artery traveling a rare route in a 91-year-old male. The common hepatic artery gave off the left hepatic, the right gastric, the gastroduodenal, and the right hepatic arteries consecutively without forming the proper hepatic artery. The right hepatic artery crossed the common bile duct anteriorly, ascended on the right side of the duct, passed the cystic duct posteriorly, and entered the right lobe of the liver. The so-called 9 o'clock artery running on the right side of the common hepatic and common biliary is reasonably speculated to be the aberrant right hepatic artery as presently shown. Developmental and clinical issues are discussed.

Ring formation of the internal iliac artery.

We observed an arterial ring communicating the superior and inferior gluteal arteries in the left half of the pelvis of an 88-year-old male. Although many previous studies have shown variations in the internal iliac artery, there has been no literature describing the fenestration. Therapeutic embolization is commonly performed for intractable bleeding in pelvic region. Surgeons should be aware of the arterial ring formation because of possible danger in the intravascular treatments. In patients with similar arterial rings, embolization of the anterior trunk of the internal iliac artery could be insufficient when blood runs through the circle of the arterial ring.

The middle suprarenal artery arising from the superior mesenteric artery.

We observed a rare case of the middle suprarenal artery branching out from the superior mesenteric artery in a 78-year-old male. This atypical artery enters the right suprarenal gland that was also supplied by the superior and the inferior suprarenal arteries as usual. In embryonic stages, vasculature of the vitelline system and the gonadal system is differentially organized. The superior mesenteric artery has been generally thought to be pure vitelline, since there has been no evidence that the superior mesenteric artery supplies other organs than digestive. We then speculate that the present middle suprarenal artery is a remnant of the embryonic gonadal artery from the superior mesenteric artery, whereas a stem artery to the testis disappeared. Surgeons should take notice of the middle suprarenal artery when operations are conducted around the superior mesenteric artery.

High range resolution ultrasonographic vascular imaging using frequency domain interferometry with the Capon method.

For high range resolution ultrasonographic vascular imaging, we apply frequency domain interferometry with the Capon method to a single frame of in-phase and quadrature (IQ) data acquired using a commercial ultrasonographic device with a 7.5 MHz linear array probe. In order to tailor the adaptive beam forming algorithm for ultrasonography we employ four techniques: frequency averaging, whitening, radio-frequency data oversampling, and the moving average. The proposed method had a range resolution of 0.05 mm in an ideal condition, and experimentally detected the boundary couple 0.17 mm apart, where the boundary couple was indistinguishable from a single boundary utilizing a B-mode image. Further, this algorithm could depict a swine femoral artery with a range beam width of 0.054 mm and an estimation error for the vessel wall thickness of 0.009 mm, whereas using a conventional method the range beam width and estimation error were 0.182 and 0.021 mm, respectively. The proposed method requires 7.7 s on a mobile PC with a single CPU for a 1×3 cm region of interest. These findings indicate the potential of the proposed method for the improvement of range resolution in ultrasonography without deterioration in temporal resolution, resulting in enhanced detection of vessel stenosis.

Immunohistochemical detection of L-DOPA-derived dopamine within serotonergic fibers in the striatum and the substantia nigra pars reticulata in Parkinsonian model rats.

On the basis of our previous studies in the normal rat [Arai, R., Karasawa, N., Geffard, M., Nagatsu, I., 1995. L-DOPA is converted to dopamine in serotonergic fibers of the striatum of the rat: a double-labeling immunofluorescence study. Neurosci. Lett. 195, 195-198; Arai, R., Karasawa, N., Nagatsu, I., 1996a. Aromatic L-amino acid decarboxylase is present in serotonergic fibers of the striatum of the rat. A double-labeling immunofluorescence study. Brain Res. 706, 177-179; Arai, R., Karasawa, N., Nagatsu, I., 1996b. Dopamine produced from L-DOPA is degraded by endogenous monoamine oxidase in neurons of the dorsal raphe nucleus of the rat: an immunohistochemical study. Brain Res. 722, 181-184] we have assumed that exogenously administered L-dihydroxyphenylalanine (L-DOPA) is converted into dopamine (DA) in serotonergic (5-HT) fibers within the striatum (ST) and the substantia nigra pars reticulata (SNR). In the present study, an attempt was made to confirm the assumptions in Parkinsonian rats, which were produced by unilateral injections of 6-hydroxydopamine (6-OHDA) into the substantia nigra pars compacta (SNC). The rats exhibiting more than 150 total controversial circles were regarded as satisfactory models of Parkinson disease (PD). Using a dual immunofluorescence histochemistry, we examined DA-immunoreactivity in the 5-HT fibers within the ST and the SNR of the PD model rats after L-DOPA was injected intraperitoneally. In experimental cases with the L-DOPA administration, DA-immunoreactivity was detected in 5-HT fibers in both the ST and the SNR on the 6-OHDA injection side; no DA-immunoreactivity was found in 5-HT fibers in the ST or the SNR in control cases without the L-DOPA administration. The results support the assumption that exogenously administered L-DOPA may be converted into DA within the 5-HT fibers in the ST and SNR of the PD model rats.

MafA differentiates rat intestinal cells into insulin-producing cells.

Recent studies have suggested that basic leucine zipper transcription factor MafA has a crucial role in pancreatic beta-cell-specific insulin gene transcription. Thus, we investigated whether MafA overexpression in the intestine induces insulin production in small-intestinal epithelial cells in vivo. Recombinant adenovirus containing MafA gene (Ad-MafA) was prepared and administered orally to streptozocin-treated diabetic rats. Insulin gene expression was observed in the intestine by RT-PCR analysis, and then insulin protein was detected by immunohistochemical analysis after Ad-MafA administration. Furthermore, MafA overexpression in the intestine increased plasma insulin levels and ameliorated hyperglycemia. These results indicate that MafA overexpression in the intestine induces intestinal epithelial cells newly to produce and release insulin.

A mammalian dpy-19 homologue is expressed in GABAergic neurons.

Gsh2 homeobox transcription factors play a crucial role in the development of GABAergic neurons. Caenorhabditis elegans's mab-5 gene is homologous to Gsh2; its expression is controlled by dpy-19. This study produced the polyclonal anti-mammalian DPY-19 (MDPY-19) antibody and showed the distribution of anti-MDPY-19 immunopositive cells. In addition, the mammalian dpy-19 (Mdpy-19) 5'-flanking region was analyzed by in vivo transient transfection assays. Mdpy-19 is expressed in ependymal cells in the adult rat brain, embryonic neuroepithelial cells, and cultured neural stem cells. In the adult rat ventricular zone, immunoreactivity with MDPY-19 of the dorsal area is stronger than that of the ventral area. Embryonic neuroepithelial cells and radial glial cells show strong anti-MDPY-19 immunoreactivity. We created the Mdpy-19 green fluorescent protein (GFP) reporter gene. Our results show that Mdpy-19 is expressed in neural stem cells and progenitor cells, especially radial glial cells. Only ependymal cells among differentiated cells express Mdpy-19 gene. However, the possibility exists that the Mdpy-19 gene is able to transcript in GABAergic neurons. It is suggested that a factor existing in mature neurons withdraws the expression of Mdpy-19.

Diet high in lipid hydroperoxide by vitamin E deficiency induces insulin resistance and impaired insulin secretion in normal rats.

To clarify the effect of dietary lipid hydroperoxide (LPO) on development of glucose intolerance, we fed Sprague-Dawley rats on a diet containing elevated LPO level for 10 weeks and measured both insulin sensitivity and insulin secretion. The contents of LPO in both plasma and skeletal muscle in the LPO-fed rats were significantly higher than those in the controls. Both insulin resistance evaluated by steady-state blood glucose (SSBG) methods and impaired insulin secretion evaluated by oral glucose tolerance test (OGTT) were found in the LPO-fed rats as compared with control rats. Furthermore, the levels of insulin receptor substrate (IRS)-1 protein in the skeletal muscle were significantly lower in the LPO-fed rats. Those impairments were not reversed in LPO-fed rats with supernormal levels of plasma vitamin E following vitamin E supplementation for 5 weeks. Moreover, the immunohistochemical study revealed that NF-kappaB-p50 protein was found in the nucleus of pancreatic beta-cells of the LPO-fed rats, whereas it was not observed in the nucleus of the islets in the control rats. These findings indicate that NF-kappaB is activated in response to oxidative stress in pancreatic islet cells in LPO-fed rats. In conclusion, our studies reveal that diet high in LPO by vitamin E deficiency accelerates glucose intolerance through impairments of both sensitivity and secretion of insulin.

Combined expression of pancreatic duodenal homeobox 1 and islet factor 1 induces immature enterocytes to produce insulin.

Immature rat intestinal stem cells (IEC-6) given the ability to express the transcription factor, pancreatic duodenal homeobox 1 (Pdx-1), yielded YK cells. Although these cells produced multiple enteroendocrine hormones, they did not produce insulin. Exposure of YK cells to 2 nmol/l betacellulin yielded BYK cells that showed the presence of insulin expression in cytoplasm and that secreted insulin into culture media. By examining the mechanism of differentiation in BYK cells, we found that another transcription factor, islet factor 1 (Isl-1) was newly expressed with the disappearance of Pax-6 expression in those cells after exposure to betacellulin. These results indicated that combined expression of Pdx-1 and Isl-1 in IEC-6 cells was required for the production of insulin. In fact, overexpression of both Pdx-1 and Isl-1 in IEC-6 cells (Isl-YK-12, -14, and -15 cells) gave them the ability to express insulin without exposure to betacellulin. Furthermore, implantation of the Isl-YK-14 cells into diabetic rats reduced the animals' plasma glucose levels; glucose levels dropped from 19.4 to 16.9 mmol/l 1 day after the injection of cells. As expected, the plasma insulin concentrations were 2.7 times higher in the diabetic rats injected with Isl-YK-14 cells compared to in controls. In summary, our results indicated that immature intestinal stem cells can differentiate into insulin-producing cells given the ability to express the transcription factors Pdx-1 and Isl-1.