Extracellular volume fraction using contrast-enhanced CT is useful in differentiating intrahepatic cholangiocellular carcinoma from hepatocellular carcinoma.

Objectives: To evaluate whether tumor extracellular volume fraction (fECV) on contrast-enhanced computed tomography (CT) aids in the differentiation between intrahepatic cholangiocarcinoma (ICC) and hepatocellular carcinoma (HCC). Methods: In this retrospective study, 113 patients with pathologically confirmed ICC (n = 39) or HCC (n = 74) who had undergone preoperative contrast-enhanced CT were enrolled. Enhancement values of the tumor (Etumor) and aorta (Eaorta) were obtained in the precontrast and equilibrium phase CT images. fECV was calculated using the following equation: fECV [%] = Etumor/Eaorta × (100 - hematocrit [%]). fECV values were compared between the ICC and HCC groups using Welch's t-test. The diagnostic performance of fECV for differentiating ICC and HCC was assessed using receiver-operating characteristic (ROC) analysis. fECV and the CT imaging features of tumors were evaluated by two radiologists. Multivariate logistic regression analysis was performed to identify factors predicting a diagnosis of ICC. Results: Mean fECV was significantly higher in ICCs (43.8% ± 13.2%) than that in HCCs (31.6% ± 9.0%, p < 0.001). The area under the curve for differentiating ICC from HCC was 0.763 when the cutoff value of fECV was 41.5%. The multivariate analysis identified fECV (unit OR: 1.10; 95% CI: 1.01-1.21; p < 0.05), peripheral rim enhancement during the arterial phase (OR: 17.0; 95% CI: 1.29-225; p < 0.05), and absence of washout pattern (OR: 235; 95% CI: 14.03-3933; p < 0.001) as independent CT features for differentiating between the two tumor types. Conclusions: A high value of fECV, peripheral rim enhancement during the arterial phase, and absence of washout pattern were independent factors in the differentiation of ICC from HCC.

Intermediate-range chemical ordering of cations in molten RbCl-AgCl.

A first sharp diffraction peak (FSDP) is observed in the X-ray total structure factor of a molten mixture of RbCl-AgCl, while both pure melts of RbCl and AgCl do not exhibit FSDP individually. Molecular dynamics simulations were performed to investigate the origin of the FSDP with the polarizable ion model (PIM). Coexistence of covalent Ag-Cl and ionic Rb-Cl bonds leads the system to evolve intermediate range ordering, which is simulated by introducing the induced polarization in different ways between Ag-Cl with fully polarizable treatment based on Vashishta-Raman potential and Rb-Cl with suppression over-polarization in the nearest neighbor contribution based on Born-Meyer potential. The partial structure factors for both the Ag-Ag and Rb-Rb correlations, SAgAg(Q) and SRbRb(Q), show a positive contribution to the FSDP, while SAgRb(Q) for the Ag-Rb correlation exhibits a negative contribution, indicating that Ag and Rb ions are distributed in an alternating manner within the intermediate-range length scale. The origin of the intermediate-range chemical ordering of cations can be ascribed to the preferred direction of the dipole moments of anions in the PIM.

Anatomical evaluation of facial nerve pathways and dissection of "premasseter space" for rhytidectomy in Asians.

BACKGROUND: Partial detachment of the superficial musculoaponeurotic system (SMAS) by dissection of the premasseter space (PMS) is an option for enhancing the effectiveness of SMAS-based rhytidectomy. The aim of this study was to identify the underlying cause of the potential risk of motor nerve impairment sometimes caused by PMS dissection and to consider the effective use of PMS dissection, especially in Asians. METHODS: Detailed dissection was carried out on six fixed Japanese cadavers to evaluate facial nerve pathways around the PMS. RESULTS: The anterior wall of the PMS was opaque because each face exhibited fibers of various thicknesses within and around the anterior border of the masseter. The ascending ramifications of the buccal trunk ran through the fibers, outside the anterior border of the masseter in some faces but within it in others. CONCLUSIONS: This study revealed the presence of a danger zone when dissecting the PMS in Asians. Severing the fibers that fix the SMAS to the masseter fascia around the anterior border of the masseter is sometimes unavoidable to attain good mobility of the SMAS. Surgeons must be mindful of the fibers near the anterior border of the masseter because they may be outside the PMS and contain buccal trunk ramifications; the anterior wall of the PMS tends to be opaque in Asians. Nonetheless, the extent of PMS dissection should be determined on an individual basis. The present findings may help to reduce relevant risks in Asian patients and standardize procedures for effective rhytidectomy. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these evidence-based medicine ratings, please refer to the table of contents or the online instructions to authors at www.springer.com/00266.

Petrous apex cholesterol granuloma treated via the endoscopic transsphenoidal approach.

Numerous surgical approaches have been used to treat petrous apex cholesterol granulomas. They are usually treated via the transtemporal- or middle fossa approach; some are managed endoscopically. We present a patient treated by the endoscopic transsphenoidal approach and review the literature.

Therapeutic importance of transplantation of a vascularised free flap for an oromandibular injury with infection and severe scar contracture.

We report three patients who had transplantation of vascularised free flaps, peroneal osteocutaneous flap in two patients and radial forearm flap in one patient, for the treatment of oromandibular injuries that had not healed as a result of infection after initial treatment.

Analysis of estrogenic activity of foodstuffs and cigarette smoke condensates using a yeast estrogen screening method.

Hormone mimics present in our environment are of concern because such agents could potentially reduce fertility and increase sexual dysfunction in wildlife and increase the risk of breast and reproductive organ cancers in man. Therefore, monitoring of the levels of estrogenic compounds in environmental materials is essential in order to prevent their exposure to man and to discover potential harmful effects on human health. In the present study, we analyzed estrogenic activity in 23 foodstuffs and cigarette smoke condensate samples extracted with an organic solvent, using the yeast estrogen screening (YES) system. Three soybean-related foodstuffs (soy sauce, tofu, miso), beer, coffee and cigarette smoke condensates showed clear estrogenic activity in the YES system. HPLC fractionations followed by the YES of these YES-positive samples revealed the presence of many estrogenic compounds in cigarette smoke condensates, whereas the other samples exerted estrogenic activities in only one or two fractions. Genistein was able to be isolated as the major active principle in soy sauce, tofu and miso, its concentration in these three foodstuffs ranging from 0.1 to 394 microg/g or ml. 8-Prenylnaringenin was also isolated from beer extracts as a major compound with estrogenic activity present at 0.22-4.0 ng/ml. Estrogenic activity of 8-prenylnaringenin with YES was 10-times as high as that of genistein, although it was 100-times less than that of 17beta-estradiol. Based on our results in vitro, 10 mg miso and 10 ml beer can be calculated to have similar estrogenic activity to 1 pmole 17beta-estradiol. It is very important that the effects of genistein and 8-prenylnaringenin on human health are elucidated.

Immunohistochemical analysis of RCAS1 in human pituitary adenomas.

It has been reported that RCAS1 (receptor-binding cancer antigen expressed on SiSO cells) acts as a ligand for a receptor present on normal peripheral lymphocytes and induces apoptotic cell death. It is expressed in uterine and ovarian carcinomas, especially in invasive cancers. This immunohistochemical study is aimed to elucidate the expression of RCAS1 in human pituitary adenomas in order to clarify its role in their proliferative regulation and invasiveness. Five normal pituitary glands, 50 human pituitary adenomas, and one malignant glioma were subjected to immunohistochemical studies. In normal pituitary glands, immunostaining of RCAS1 and MIB-1 was not found. In malignant glioma, large numbers of cell nuclei were positive for MIB-1 (MIB-1 index: 28%), and RCAS1 was detected both in the cytoplasm and on the membrane of the tumor cells. Expression of RCAS1 was noted in 48% of pituitary adenomas immunohistochemically (60.0% of growth hormone-secreting adenomas, 60.0% of prolactin-secreting adenomas, 42.9% of adrenocorticotrophin-secreting adenomas, 40.0% of thyroid-stimulating hormone-secreting adenomas, 33.3% of nonfunctioning adenomas, and 44.4% of gonadotropin-subunit-positive adenomas). It showed no correlation with tumor type, size, and invasiveness. The statistically significant relationship between RCAS1 and MIB-1 positivity was identified in our study. These results suggest that expression of RCAS1 as well as MIB-1 positivity predict the growth potential of individual pituitary adenomas.

Potassium channel blocker activates extracellular signal-regulated kinases through Pyk2 and epidermal growth factor receptor in rat cardiomyocytes.

OBJECTIVES: We sought to determine whether potassium (K(+)) channel blockers (KBs) can activate extracellular signal-regulated kinase (ERK) and to characterize the upstream signals leading to ERK activation in cardiomyocytes. BACKGROUND: Because KBs attenuate K(+) outward current, they may possibly prolong the duration of action potentials, leading to an increase in calcium (Ca(2+)) transient ([Ca(2+)](i)) in cardiomyocytes. Elevation of intracellular Ca(2+) levels can trigger various signaling events. Influx of Ca(2+) through L-type Ca(2+) channels after membrane depolarization induced activation of MEK and ERK through activation of Ras in neurons. Although KBs are frequently used to treat cardiac arrhythmias, their effect on signaling pathways remains unknown. METHODS: Primary cultured rat cardiomyocytes were stimulated with four different KBs-4-aminopyridine (4-AP), E-4031, tetra-ethylammonium and quinidine-and phosphorylation of ERK, proline-rich tyrosine kinase 2 (Pyk2) and epidermal growth factor receptor (EGFR) was detected. Action potentials were recorded by use of a conventional microelectrode. (Ca(2+))(i) was monitored by the fluorescent calcium indicator Fluo-4. RESULTS: E-4031, 4-AP, tetra-ethylammonium and quinidine induced phosphorylation of ERK. 4-Aminopyridine prolonged the duration of action potentials by 37% and increased (Ca(2+))(i) by 52% at 1 mmol/l. Pre-incubation of ethyleneglycoltetraacetic acid, 1,2-bis(2-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid tetrakis and diltiazem completely blocked this phosphorylation, whereas flufenamic acid and benzamil did not. 4-Aminopyridine induced tyrosine phosphorylation of Pyk2 and EGFR, which peaked at 5 and 10 min, respectively. Cytochalasin D, AG1478 and dominant-negative EGFR strongly inhibited the phosphorylation of ERK, whereas calphostin C, calmidazolium and KN62 did not. CONCLUSIONS: These findings indicate that KBs induce ERK activation, which starts with Ca(2+) entry through the L-type Ca(2+) channel in cardiomyocytes, and that EGFR and Pyk2 are involved in this activation.

Immunohistochemical analysis of p27 (Kip1) in human pituitary glands and in various types of pituitary adenomas.

p27 (Kip1) plays regulatory roles in the cell cycle by inhibiting the activity of cyclin dependent kinases (CDKs). This immunohistochemical study is aimed at elucidating the expression of p27 in human pituitary and in various types of pituitary adenomas in order to clarify its role in the regulation of proliferation. Sixteen normal pituitary glands and 179 human pituitary adenomas were used for immunohistochemical studies. The tissues were fixed in 10% formalin and embedded in paraffin. Indirect peroxidase method was performed after heat-induced antigen retrieval using a monoclonal antibody against p27 protein. p27 protein was expressed in the nuclei of all 16 normal human pituitary glands. p27 protein was also expressed in 128 of 179 cases of pituitary adenomas (71.5%). A marked decrease of p27 expression was noted in ACTH-secreting adenomas, 8/20 (40.0%), compared with other types of pituitary adenomas--GH-secreting adenomas, 35/46 (76.1%); PRL-secreting adenomas, 22/33 (66.7%); TSH-secreting adenomas, 8/11 (72.7%); and nonfunctioning adenomas, 55/69 (79.7%). These results suggest that p27 may play some role in the regulation of proliferation in all types of pituitary adenomas. The lower levels of p27 in ACTH-secreting adenoma is of particular interest with respect to the intermediate lobe-derived pituitary tumor developed in p27 knockout mice.

The third naturally occurring attractant toward zoospores of phytopathogenic Aphanomyces cochlioides from the Spinacia oleracea host plant.

A bioassay-guided survey of spinach leaf constituents resulted in 5,4'-dihydroxy-3,3'-dimethoxy-6,7-methylenedioxyflavone being identified as the third naturally-occurring attractant in the host plant toward the zoospores of its pathogen, Aphanomyces cochlioides. The isolate showed attracting activity around Chromosorb W AW particles (60-80 mesh) coated with a 10(-5) M solution in a zoospore suspension. However, this activity was 1/100-1/1000 less than that of cochliophilin A, an attractant in the roots of spinach. Bioassays with the present isolate and related compounds revealed that 5,3',4'-trihydroxy-3-methoxy-6,7-methylenedioxyflavone did not possess attractant activity, but rather weak antagonistic activity toward the former two attractants from spinach.

NMR determination of the absolute configuration of a macrophomate synthase inhibitor by using an axial chiral reagent.

Macrophomate synthase catalyzes an extraordinary four-step transformation from oxalacetate and 2-pyrone to macrophomic acid by an intermolecular Diels-Alder reaction. The absolute configuration of the most potent macrophomate synthase inhibitor; (-)-2-carboxylmethyl-1-methoxybicyclo[2.2.2]oct-5-ene-2-carboxylic acid, was determined to be (1S, 2R, 4R) by using an axial chiral reagent.

Accumulation of oxidative DNA damage, 8-oxo-2'-deoxyguanosine, and change of repair systems during in vitro cellular aging of cultured human skin fibroblasts.

Effects of in vitro cellular aging on the content of 8-oxo-2'-deoxyguanosine, a typical oxidation product of DNA bases, were examined in cultured human skin fibroblasts. The 8-oxo-2'-deoxyguanosine content in the DNA of TIG-3S cells established from skin tissues of a fetal donor increased immediately before the cessation of proliferation. TIG-114 and TIG-104 cells established from skin tissues of adult and aged donors, respectively, showed similar changes in 8-oxo-2'-deoxyguanosine content during in vitro cellular aging. The accumulation of 8-oxo-2'-deoxyguanosine in late-passage cells was dependent on the number of cell divisions, and not on the cultivation time. Increases in the activities of superoxide dismutase and glutathione peroxidase were observed prior to the increase in 8-oxo-2'-deoxyguanosine content, while the catalase activity decreased gradually during in vitro cellular aging at late-passage. Furthermore, the activities of 8-oxo-2'-deoxyguanosine endonuclease and DNA polymerases decreased with the progression of proliferation. These results indicate that defense systems against oxidative stress in late-passage cells remain sufficiently active before the cessation of cell division, but that repair systems against oxidative damage decay at late-passage. Oxidative stress beyond the antioxidant capacity and/or repair activity seems to result in an accumulation of 8-oxo-2'-deoxyguanosine in late-passage cells.

Chemotaxis of fungal zoospores, with special reference to Aphanomyces cochlioides.

Zoospores of phytopathogenic fungi accumulate at the potential infection sites of host roots by chemotaxis. The aggregated spores then adhere, encyst, germinate, and finally penetrate into the root tissues to initiate infection. Some of the host-specific attractants have already been identified. The host-specific attractants also induce cell differentiation of certain zoospores under laboratory conditions. This indicates that a signal released from the roots of the host plant guides the pest propagules for orientation and prepares them for establishing a host-pathogen relationship by necessary physiological changes. Some non-host plant secondary metabolites were found to markedly regulate behavior and viability of zoospores, suggesting that non-host compounds may also play a role in protecting the non-host plants from the attack of zoosporic fungi. We hypothesized that zoospores perceive the host signal(s) by specific G-protein-coupled receptors and translate it into responses by way of the phosphoinositide-Ca2+ signaling cascade. The details of the signal transduction mechanism in fungal zoospores are yet to be discovered. In this report, we review the signaling and communications between phytopathogenic fungal zoospores and host and non-host plants with special reference to Aphanomyces cochlioides.

Capability of wild Rosa rugosa and its varieties and hybrids to produce sesquiterpene components in leaf glandular trichomes.

The sesquiterpene contents in leaves of wild Rosa rugosa and of sixty-one hybrid rugosas were quantitatively measured by a GC analysis. In this group of samples, the greater the number of glandular trichomes the hybrid rugosas possessed on their leaves, the larger the amount of sesquiterpenes they accumulated. In contrast, those having no leaf glandular hairs contained only a trace amount of sesquiterpene components. The concentrations of bisaborosaol A (1) and carota-1,4-dienaldehyde (2) as representative sesquiterpenes of R. rugosa were positively correlated with the density of the glandular trichomes. Furthermore, an approximately regular correlation was observed between the concentrations of 1 and 2 in most of the sesquiterpene-producing hybrid rugosas, regardless of their productivity. This suggests that a major part of these hybrid rugosas have inherited from R. rugosa the ability to produce two skeletally different sesquiterpenes in parallel with a phenotype to develop leaf glandular trichomes. This investigation also led to discovering 1-dominant (e.g., Amelie Gravereaux and Purple Pavement), 2-dominant (e.g., David Thompson), and other-dominant (e.g., Martin Frobisher) types of sesquiterpene-producing hybrid rugosas.

Antimicrobial cuparene-type sesquiterpenes, enokipodins C and D, from a mycelial culture of Flammulina velutipes.

Two new cuparene-type sesquiterpenes, enokipodins C (1) and D (2), were isolated from culture medium of an edible mushroom, Flammulina velutipes, along with enokipodins A (3) and B (4). The structures of 1 and 2 were determined using spectroscopic methods (HRMS, (1)H and (13)C, and 2D NMR). The absolute configuration of enokipodin C was determined from the observed (1)H NMR chemical shifts and NOEs in NOESY experiments after conversion into the corresponding esters with the chiral reagent 2-(2'-methoxy-1'-naphthyl)-3,4-dichlorobenzoic acid. All the metabolites showed antimicrobial activity against a fungus, Cladosporium herbarum, and Gram-positive bacteria, Bacillus subtilis and Staphylococcus aureus.

Functional reconstruction of mobile tongue and suprahyoid muscles after resection of cancer of the tongue.

The mobile tongue and the suprahyoid muscles were reconstructed in a man who had had a total resection of the mobile tongue with the oral floor and bilateral radical neck dissection (extensive resection of the bilateral suprahyoid muscles) for cancer of the tongue. A rectus abdominis musculocutaneous flap with the tenth intercostal nerve anastomosed to the remaining hypoglossal nerve was used for the reconstruction. The rectus abdominis muscle was inserted between the mandible and the hyoid bone, taking account of the anatomical positions of the tongue and suprahyoid muscles; the rectus sheath was then firmly sutured to the mandible and hyoid bone. This procedure created firm tendinous insertions between the mandible and hyoid bone. After the operation, the reconstructed tongue showed no sign of atrophy, and the reconstructed supra-hyoid muscles (rectus abdominis muscle) could be moved, and the hyoid bone could be raised.

Rare variant of the intrasoleus musculocutaneous perforator: clinical considerations in raising a free peroneal osteocutaneous flap.

To improve the stability of the cutaneous portion of the vascularized free peroneal osteocutaneous flap (the so-called peroneal flap), various anatomic studies have been conducted, and a reliable notion of the course of cutaneous perforators has been provided; however, anatomic anomalies have occasionally been observed. The authors encountered a rare type of cutaneous perforator in the distal third of the lower leg. It was located under the inferior surface of the flexor hallucis longus muscle, after running in the soleus muscle parallel to the fibula, and then joined the posterior tibial artery. Since absolutely definite diagnostic methods have not been established for the course of the cutaneous perforator, surgery with the possibility of secondary anastomosis, always with a separate peroneal flap, should be a consideration.

Repellent activity of estrogenic compounds toward zoospores of the phytopathogenic fungus Aphanomyces cochlioides.

Screening chemical compounds, we found that a xenoestrogen, bisphenol A, showed potent repellent activity against the zoospores of Aphanomyces cochlioides. Based on this finding, we tested a number of androgenic and estrogenic compounds (e.g. testosterone, progesterone, estradiols, diethylstilbestrol, estrone, estriol, pregnenolone, dienestrol etc.) on the motility behavior of A. cochlioides zoospores. Interestingly, most of the estrogenic compounds exhibited potent repellent activity (1 microg/ml or less by the "particle method") toward the motile zoospores of A. cochlioides. We derivatized some of the estrogens and discussed the relationship between the structure of active molecules and their repellent activity. Apparently, aromatization of the A ring with a free hydroxyl group at C-3 position of a steroidal structure is necessary for higher repellent activity. Interestingly, methylation of diethylstilbestrol (DES) yielded completely different activity i.e. both mono- and di-methyl ethers of DES showed attractant activity. Moreover, the attracted zoospores were encysted and then germinated in the presence of di-methyl ether of DES. The potential usefulness of this repellent test is discussed for the detection of estrogenic activity of naturally occurring compounds, and the possible role of phytoestrogens in host/parasite interactions. So far, this will be the first report of repellent activity of estrogenic compounds toward trivial fungal zoospores.

Single bout of exercise eliminates the immobilization-induced oxidative stress in rat brain.

We were interested in the effects of immobilization (IM), a single bout of exercise (E) and immobilization followed by exercise (EIM) on memory and oxidative damage of macromolecules in hippocampus of rat brain. Eight hours of IM resulted in impairment of passive avoidance test (memory retrieval deficit) and increased latency to start locomotion in an open-field test. Two hours of swimming did not significantly alter the memory retrieval deficit and latency, while the EIM group had longer latency and similar memory than control and E groups. The oxidative damage of lipids, proteins and nuclear DNA increased significantly in IM group and no increase was observed in E and EIM animals. The activity of proteasome was not altered in any groups. The activity of glutamine synthetase (GS) was decreased in IM group (P < 0.05), this down regulation was not observed in E and EIM groups. These data suggest that oxidative damage of macromolecules is associated with impaired cognitive function. Single bout of exercise after immobilization eliminates the oxidative damage of macromolecules and normalizes memory function, probably by its ability to restore the activity level of GS and eliminate the consequences of immobilization-induced prolonged efflux of glutamate.