Venous drainage of the face.

The venous anatomy of the face was examined in 12 fresh cadavers. Venograms and arteriovenograms were obtained after the injection of contrast medium. In 8 of the 12 cadavers, a large loop was formed by the facial vein, the supratrochlear vein, and the superficial temporal vein, which became the main trunk vein of the face. In 4 of the 12 cadavers, the superior lateral limb of the loop vein was less well developed. The loop vein generally did not accompany the arteries of the face. Cutaneous branches of the loop vein formed a polygonal venous network in the skin, while communicating branches ran toward deep veins. These findings suggest that blood from the dermis of the face is collected by the polygonal venous network and enters the loop vein through the cutaneous branches, after which blood flows away from the face through the superficial temporal vein, the facial vein, and the communicating branches and enters the deep veins.

Population dynamics of Armigeres subalbatus (Diptera: Culicidae) across a temperate altitudinal gradient.

Understanding the impacts of weather fluctuations, and environmental gradients, on the abundance of vectors is fundamental to grasp the dynamic nature of the entomological risk for disease transmission. The mosquito Armigeres subalbatus (Coquillet) is a common vector of filariasis. Nevertheless, its population dynamics have been relatively poorly studied. Here, we present results from a season long study where we studied spatio-temporal abundance patterns of Ar. subalbatus across the altitudinal gradient of Mt. Konpira in Nagasaki, Japan. Spatially, we found that abundance of adult Ar. subalbatus decreased with altitude and increased in areas where the ground was rich in leaf litter. Similarly, adult activity was observed only when relative humidity was over 65%. Temporally, we found that peaks in abundance followed large rainfall events. Nevertheless, this mosquito was under significant density dependence regulation. Our results suggest that Ar. subalbatus population peaks following large rainfall events could reflect the recruitment of individuals that were dormant as dry eggs. We did not find a clear signal of temperature on abundance changes of this mosquito, but only on its phenology. Since ground cover seemed more critical than temperature to its spatial distribution, we propose that this mosquito might have some degree of autonomy to changes in temperature.

Phase stability of a garnet-type lithium ion conductor Li7La3Zr2O12.

The phase stability of Li7La3Zr2O12 (LLZ) was investigated using high temperature X-ray diffraction (HT-XRD). An Al-free tetragonal LLZ phase transformed into a non-quenchable cubic phase around 650 °C. The phase transformation process between the tetragonal phase and the new cubic phase showed perfect reversibility. The thermal analysis showed a pair of endothermic and exothermic peaks around 640 °C that is in good agreement with the phase transformation process observed in the HT-XRD study. The non-quenchable high temperature cubic phase showed high ionic conductivity with extraordinarily low activation energy (0.117 eV). The tetragonal phase showed another phase transformation to a low temperature (LT) cubic phase around 150-200 °C in air by absorbing CO2 into the structure. The preferred temperature for the CO2 absorption process was around 200 °C and the absorbed CO2 was extracted once the temperature reached 450 °C or above resulting in the phase transformation back to the tetragonal phase. On the other hand the high temperature (HT) cubic phase which shows high ionic conductivity was stabilized by Al substitution. A Li-poor LLZ containing impurity phases such as La2Zr2O7 and La2O3 effectively reacted with γ-Al2O3 resulting in the formation of a pure Al-stabilized cubic LLZ, while the stoichiometric LLZ took a much longer time to complete the Al-substitution. The result suggested that the formation of Li vacancies is the primary step in the formation of the Al-stabilized cubic phase.

Evaluation of the effects of freeze-dried soybean curd intake on cholesterol levels using a novel biomarker.

To evaluate the effects of freeze-dried soybean curd intake on serum cholesterol levels, we examined the subclasses of cholesterol for healthy adult volunteers who continued to eat a piece of freeze-dried soybean curd each day along with their ordinary diet for four weeks. Of 12 subjects, the soybean curd diet proved effective in 2 cases; small dense LDL (sd-LDL) cholesterol levels were significantly reduced in association with the decrease in LDL cholesterol levels. These results suggested that daily intake of freeze-dried soybean curd may lead to an improvement in cholesterol metabolism in subjects with originally higher cholesterol levels and that sd-LDL cholesterol can be a novel biomarker for evaluation of the cholesterol lowering-action of functional food.

Pharmacogenomics-based clinical studies using a novel, fully automated genotyping system.

Clinical investigations into single nucleotide polymorphisms (SNPs) in drug metabolism have already been set out for clinical trials in subject groups classified as extensive metabolizers or poor metabolizers. In particular, the frequency of CYP2C19 in poor metabolizers within the Japanese population is relatively high, and genetic variations result in differences in kinetics and pharmacological action, e.g. clinical response to proton pump inhibitors which are mainly metabolized by 2C19 in the liver. We introduced a novel, fully automated genotyping system and used it in the genotyping of CYP2C19. The completed system is based on the analysis of a melting curve of probe DNA which is bound to the target SNP site using a fluorescence quenching probe. The system enables automated and multiple SNP-genotyping from sample preparation. This fully automated system of analysis can be adapted to clinical studies, e.g. classification of genes related to pharmacokinetics and target receptors by genetic variations.

Rapid, severe repigmentation of congenital melanocytic naevi after curettage and dermabrasion: histological features.

BACKGROUND: Curettage and dermabrasion are effective in treating giant congenital melanocytic naevi (GCMN). We report two patients with rapid, severe postoperative repigmentation. To the best of our knowledge this is the first report on the histological features of such patients. OBJECTIVES: We wish to call attention to histological features that may cause rapid, severe repigmentation after curettage and dermabrasion of medium to giant CMN. PATIENTS/METHODS: From 1998 to 2002, we treated 23 patients with medium to giant CMN with curettage and dermabrasion. Patients being treated ranged in age from 1 month to 19 years. Histological samples were taken from the centre of naevi in all patients during surgery. Histological types were 12 intradermal and 11 compound. Follow-up after curettage lasted at least 3 years. RESULTS: Among our 23 patients only two showed repigmentation soon after surgery. Histological sections from these two patients indicated naevoid cells in the deep dermis along hair follicles or sebaceous glands. However, no such pigmented naevoid cells along hair follicles were observed in samples from patients successfully treated with curettage and dermabrasion with less repigmentation. CONCLUSIONS: Although we saw only two cases of repigmentation soon after curettage and dermabrasion, we suspect a correlation between pigmented naevoid cells around hair follicles and repigmentation. If histological sections of skin biopsies show pigmented cells along hair follicles in the deep dermis, other treatments such as total skin resection followed by skin grafting or tissue expansion may be better choices than curettage or dermabrasion.

Inhibitory effect of cinnamaldehyde, derived from Cinnamomi cortex, on the growth of influenza A/PR/8 virus in vitro and in vivo.

We have investigated the inhibitory effect of trans-cinnamaldehyde (CA), one of the principal constituents of essential oil derived from Cinnamomi cortex, on the growth of influenza A/PR/8 virus in vitro and in vivo. When 1-h drug treatment was initiated at various times post-infection (p.i.) in Madin-Darby canine kidney cells using a fixed dose of CA (40 microM), the maximum inhibitory effect (29.7% virus yield of control) was obtained when drug treatment was started at 3h p.i. Under the same treatment schedule, CA inhibited the virus growth in a dose-dependent manner (20-200 microM), and, at 200 microM, the virus yield was reduced to an undetectable level. RT-PCR and SDS-PAGE analyses showed that CA inhibited viral protein synthesis at the post-transcriptional level. In mice infected with the lung-adapted PR-8 virus, inhalation (50mg/cage/day) and nasal inoculation (250 microg/mouse/day) of CA significantly increased survival rates on the 8 days to 100% and 70%, respectively, in contrast to a survival rate of 20% in the untreated control group. Importantly, inhalation of CA caused virus yield reduction by 1 log in bronchoalveolar lavage fluid on day 6 after infection, compared with that of the untreated control group. These findings might provide further support to the empirical indication of Cinnamomi cortex-containing Kampo medicines for acute respiratory infectious diseases.

Anatomical relationship between arteries and veins in the paraumbilical region.

Veins in the paraumbilical region were investigated in eight fresh cadavers, in which radiopaque materials were injected into both the arterial and the venous systems, to determine their locational relationship to the arteries. Veins in the skin and subcutaneous tissue consisted of venae comitantes and non-venae comitantes. The main trunk of the non-venae comitantes was the superficial inferior epigastric vein, and it formed a polygonal venous network in the skin layer. A large communicating vein, which did not accompany an artery, connected the venous network to a vena comitans of a large paraumbilical arterial perforator. Venous blood that had perfused the dermis of the paraumbilical region had two kinds of pathways to a deep vein: through the superficial inferior epigastric vein to the femoral vein and through the vena comitans to the deep inferior epigastric vein.

Anatomical study of the venous drainage architecture of the scapular skin and subcutaneous tissue.

Venous anatomy of the skin and subcutaneous adipofascial tissue in the scapular region was examined in 14 specimens of 12 fresh cadavers that had been injected systemically with contrast medium. Three-dimensional analysis was performed by radiographing the specimens stereoscopically and splitting them into the skin and subcutaneous adipofascial tissue layers. From the architecture, most of the venous blood that had perfused the dermis was considered to pool in a polygonal venous network, located in the skin layer; to flow chiefly through some large communicating veins; and to enter the scapular, parascapular, or circumflex scapular veins. Most of the venous blood that had perfused the subcutaneous adipofascial tissue was considered to enter the scapular or parascapular veins directly.

Anatomical relationship between arteries and veins in the scapular region.

Veins in the scapular region were investigated in five fresh cadavers, in which radiopaque materials were injected into both the arterial and the venous systems, to determine their locational relationship to the arteries. This radiographic technique is very useful for comparing veins and arteries. Many veins not accompanying arteries were observed, and veins in the skin and subcutaneous tissue were considered to consist of venae comitantes and non-venae-comitantes. The non-venae-comitantes formed a mainly polygonal venous network in the skin layer, and large communicating veins connected the venous network to the scapular or parascapular veins. These veins played a role in the drainage of venous blood that had perfused the dermis and, in this sense, they should be named cutaneous veins.

Inhibitory effect of (+)-catechin on the growth of influenza A/PR/8 virus in MDCK cells.

We investigated whether (+)-catechin, a building block of tannins contained in the extract of Ephedrae herba (EHext), exerts an inhibitory effect on the acidification of intracellular compartments such as endosomes and lysosomes (referred to as ELS), and thereby inhibits the growth of influenza A PR/8/34 (PR8) virus (H1N1 subtype) in Madin-Darby canine kidney cells. The vital fluorescence microscopic study with acridine orange showed that 1-h treatment with (+)-catechin inhibited the acidification of ELS in a concentration-dependent manner (1.0-10.0 mM). Moreover, the growth of PR8 virus was inhibited markedly when the cells were treated with (+)-catechin (1.25-10.0 mM) for 1 h immediately after infection, or treated within as little as 5 to 10 min after infection. Conversely, virus growth resumed within 3 h concomitantly with the reappearance of acidified ELS after removal of (+)-catechin. Similar to EHext, (+)-catechin inhibited both the acidification of ELS and the influenza virus growth. It suggests that (+)-catechin is one of the active components in EHext.

Sialyl Lewis X and anti-P-selectin antibody attenuate lipopolysaccharide-induced acute renal failure in rabbits.

BACKGROUND/AIM: Acute renal failure (ARF) is one of the common problems associated with sepsis or multiple organ dysfunction syndrome (MODS). We have investigated the effects of inhibiting selectin-mediated cell adhesion on lipopolysaccharide-induced ARF in rabbits, using sialyl Lewis X oligosaccharide and PB1.3, an anti-human P-selectin monoclonal antibody, as inhibitors. METHODS: ARF was induced by intravenous administration of lipopolysaccharide (0.3 mg/kg, i.v. bolus injection) to New Zealand White rabbits. Induction of ARF was characterized by increases in blood urea nitrogen (BUN), creatinine, and the number of polymorphonuclear leukocytes infiltrating glomeruli, and by fibrin deposition in glomeruli, and tubular dilatation in the kidney. Sialyl Lewis X oligosaccharide (14 mg/kg, i.v. bolus injection immediately before lipopolysaccharide administration and 9 mg/kg/h, i.v. infusion) or PB1.3 (5 mg/kg, i.v. bolus injection before lipopolysaccharide administration), anti-P-selectin antibody, were treated. RESULTS: Treatment with sialyl Lewis X oligosaccharide inhibited the increases in BUN, creatinine, and the number of infiltrating polymorphonuclear leukocytes, and attenuated histopathological impairments. Similarly, treatment with PB1.3 prevented some of the characteristics associated with lipopolysaccharide-induced ARF, not but the increase in creatinine. CONCLUSION: These results suggest that selectin inhibitors, including sialyl Lewis X oligosaccharide and PB1.3, may provide clinical benefits in the prevention of ARF associated with sepsis and MODS. To our knowledge, this is the first report that P-selectin is directly involved in lipopolysaccharide-induced ARF in rabbits.

Nitric oxide-mediated antitumor activity induced by the extract from Grifola frondosa (Maitake mushroom) in a macrophage cell line, RAW264.7.

We have investigated D-fraction (MDF) extracted from Grifola frondosa (Maitake mushroom) on the inducible nitric oxide synthase (iNOS)-mediated nitric oxide (NO) production in RAW264.7 (RAW) cells, a murine monocyte/macrophage cell line, with special reference to antitumor activity of MDF against human hepatoma-derived huH-1 cells. MDF could induce iNOS mRNA expression in RAW cells in a dose range of more than 30 microg/ml, but the effect of 10 microg/ml of MDF was negligible. The iNOS mRNA expression induced by 100 microg/ml of MDF was 6 hrs later, but lasted for a longer time than that of lipopolysaccharide (LPS), a representative iNOS inducer. Although iNOS mRNA levels in MDF-stimulated cells were almost equal to LPS-stimulated cells at the peak time, the cumulative amount of nitrite was only about 50% compared with that of LPS-treated cells. When huH-I cells were cultured in MDF containing media in a 24-well plate with inserted porous bottom in the presence or absence of RAW cells, the viability of huH-1 cells decreased significantly only in the presence of RAW cells in MDF dose-dependent manner. This antitumor activity of RAW cells in the presence of MDF was abolished or attenuated by the addition of L-NAME, a NOS inhibitor, confirming that this phenomenon is due to iNOS-mediated NO production by RAW cells, but not direct cytotoxic activity of MDF against huH-1 cells. These data suggest that MDF is a novel inducer for iNOS which contributes at least in part to antitumor activity of MDF.

Role of P-selectin in the migration of neutrophils to chemoattractant-induced cutaneous inflammation in mice.

The role of P selectin in the accumulation of neutrophils at acute dermal inflammatory sites induced by chemoattractants, LTB4 and IL-8 was investigated in the mouse. A mouse P-selectin-human IgG chimera bound to mouse neutrophils in vitro in a calcium-dependent manner, as detected by flow cytometry. A rat monoclonal antibody (mAb) against mouse P-selectin, RB40.34 abolished P-selectin-IgG chimera binding to mouse neutrophils, but a control antibody did not. Intradermal injection of LTB4 at a dose of 100 ng/site caused neutrophil accumulation to increase by 3-4 fold, as detected by measuring myeloperoxidase activity. Neutrophil extravasation to perivascular tissue was detected by histochemical observation. The intravenous injection of RB40.34 or the specific LTB4 antagonist, SM-15178, at doses at 5 mg/kg attenuated the accumulation of neutrophils by 55.6% and 70.3%, respectively, but a control antibody showed no effect. Similarly, intradermal administration of IL-8 at a dose of 5 microg/site induced significant neutrophil migration into the interstitial tissue of the skin, as followed by measuring myeloperoxidase activity and histopathologic analysis. The intravenous injection of RB40.34 at a dose of 5 mg/kg reduced the neutrophil accumulation by 59.2% in contrast, a control antibody showed no effect. To our knowledge, this is the first direct demonstration that P-selectin plays a substantial role in LTB4- and IL-8-induced neutrophil accumulation in mouse skin.

Anatomic study of the venous drainage architecture of the forearm skin and subcutaneous tissue.

The venous anatomy of the forearm skin was examined radiographically in 15 fresh cadavers that had been injected systemically with a lead oxide-gelatin mixture. In 10 specimens, the forearm skin was divided into the skin and superficial adipofascial layer and the deep adipofascial layer. Five specimens were radiographed stereoscopically. Despite the thinness of the skin and subcutaneous tissue of the forearm, the cutaneous vein was seen three-dimensionally. Judging from the architecture and direction of the venous valves, most of the venous blood that had perfused the dermis was believed to: (1) pool in a venous network located in the superficial zone of the skin and subcutaneous tissue, (2) flow chiefly in the accessory cephalic and median antebrachial veins, and (3) enter the cephalic and basilic veins near the antecubital fossa. Venae comitantes of the septocutaneous and musculocutaneous perforators of the radial or ulnar arteries were thought to be only bypasses to the deep vein.

Angiographic study of the subdermal plexus: a preliminary report.

The subdermal plexus was pictured angiographically in five fresh cadavers injected systemically with a lead oxide-gelatin mixture. Subdermal plexus was found either in the subdermal plane or in the deep part of the dermis. Diameters of vessels in the subdermal plexus are not uniform and there are differences in vascular continuity, which means that the subdermal plexus does not always have a random pattern. This observation is important when designing a thin flap.

The effects of delayed treatment with sialyl Lewis X against lipopolysaccharide-induced acute lung injury in rabbits.

The therapeutic effects of a selectin inhibitor against lipopolysaccharide-induced acute lung injury were studied in rabbits by using sialyl Lewis X-oligosaccharide. Lipopolysaccharide-induced acute lung injury, as characterized by an impairment of pulmonary gas exchange, clinically resembles that of the acute respiratory distress syndrome. Delayed treatments with sialyl Lewis X-oligosaccharide (55 mg kg(-1) i.v. bolus injection 0.5, 1 or 2 h after lipopolysaccharide administration+36 mg kg(-1) h(-1) i.v. infusion for 5.5, 5 or 4 h, respectively) prevented the lipopolysaccharide-induced impairments in pulmonary gas exchange, as well as the accumulation of polymorphonuclear leukocytes in the lung tissue. In contrast, this agent had no significant effects on lipopolysaccharide-induced systemic hypotension, the decrease in the number of circulating white blood cells and platelets or the decline in blood pH. This is the first demonstration that sialyl Lewis X-oligosaccharide is effective against the impairments in pulmonary gas exchange even if administered 0.5, 1 or 2 h following the lipopolysaccharide injection.

Therapeutic effect of anti-macrophage inflammatory protein 2 antibody on influenza virus-induced pneumonia in mice.

We investigated the effect of anti-macrophage inflammatory protein 2 immunoglobulin G (aMIP-2 IgG) on the progression of influenza virus-induced pneumonia in mice. When mice were infected with a mouse lung-adapted strain of influenza A/PR/8/34 virus by intranasal inoculation, neutrophil counts in the bronchoalveolar lavage fluid (BALF) increased in parallel with the kinetics of MIP-2 production, which peaked 2 days after infection. After intracutaneous injection of a dose of 10 or 100 microg of aMIP-2 IgG once a day on days 0 and 1, neutrophil counts in BALF on day 2 were reduced to 49 or 37%, respectively, of the value in the control infected mice administered anti-protein A IgG. The antibody administration also improved lung pathology without affecting virus replication. Furthermore, by prolonged administration with a higher or lower dose for up to 5 days, body weight loss became slower and finally 40% of mice in both treatment groups survived potentially lethal pneumonia. These findings suggest that MIP-2-mediated neutrophil infiltration during the early phase of infection might play an important role in lung pathology. Thus, MIP-2 was considered to be a novel target for intervention therapy in potentially lethal influenza virus pneumonia in mice.

Protective effect of anti-P-selectin monoclonal antibody in lipopolysaccharide-induced lung hemorrhage.

Excessive leukocyte accumulation is involved in the pathogenesis of the sepsis-induced acute lung injury. Selectins are essential to the interaction between leukocytes and endothelial cells. In this report, we investigated the role of selectins in the severe lung injury induced by lipopolysaccharide (LPS). Significant lung hemorrhage was observed 24 h after the intravenous administration of LPS (1 mg/kg). First, we evaluated the effect of sialyl Lewis X-oligosaccharide (SLeX-OS), a derivative of sialyl Lewis X which is one of the ligands for E-, P- and L-selectins. The treatment with SLeX-OS (26.5 mg/kg iv bolus + 19.8 mg/kg iv infusion) resulted in a decrease of lung hemorrhage by 49.5% (P<0.05 versus the control group). Second, we tested the effect of anti-P-selectin monoclonal antibody (MAb), PB 1.3, to investigate the role of P-selectin. The bolus administration of PB1.3 at a dose of 5 mg/kg attenuated the lung hemorrhage by 74.6% (P<0.05 versus the control group). In addition, we also detected an increase of soluble P-selectin in plasma 24 h after the injection of LPS. These results suggest that P-selectin has a substantial role in the pathogenesis of the lung injury induced by LPS.

Protective effects of sialyl Lewis X and anti-P-selectin antibody against lipopolysaccharide-induced acute lung injury in rabbits.

The prophylactic effects of selectin inhibitors on lipopolysaccharide-induced acute lung injury were studied in rabbits by using sialyl Lewis X-oligosaccharide and PB1.3, an anti-human P-selectin monoclonal antibody. Lipopolysaccharide-induced acute lung injury resembles that of the acute respiratory distress syndrome, in which there is a decrease in arterial blood oxygen tension (PaO2) and an increase in the difference between alveolar and arterial oxygen tension (A-aDO2). Prophylactic treatment with the selectin inhibitors, sialyl Lewis X-oligosaccharide (55 mg kg(-1) i.v. bolus injection immediately before lipopolysaccharide administration + 36 mg kg(-1) h(-1) i.v. infusion for 4 h) and PB1.3 (5 mg kg(-1) i.v. bolus injection immediately before lipopolysaccharide administration), prevented the lipopolysaccharide-induced impairments in pulmonary gas exchange. In contrast, these agents had no significant effects on lipopolysaccharide-induced systemic hypotension, the decrease in the number of circulating white blood cells and platelets, the decline in blood pH, or the increase in arterial CO2 tension (PaCO2). These results indicate that selectin inhibitors including sialyl Lewis X-oligosaccharide and the anti-P-selectin antibody, PB1.3, attenuate lipopolysaccharide-induced acute lung injury in rabbits. This is the first demonstration that P-selectin is directly involved in the development of lipopolysaccharide-induced impairments in pulmonary gas exchange.