Cl- channels regulate lipid droplet formation via Rab8a expression during adipocyte differentiation.

Several studies have shown that Cl- channels regulate the differentiation of some cell types. Thus, we investigated the role of Cl- channels on adipocyte differentiation using adipose tissue-derived stem cells (ASCs) and Cl- channel blocker. We induced rabbit ASCs into adipocytes using Cl- channel blocker. The expression levels of adipocyte markers were no significant difference between the cells treated with a Cl- channel blocker NPPB and untreated cells. However, when the cells were treated with NPPB, lipid droplets (LDs) sizes decreased compared with the untreated control. Interestingly, the expression levels of Rab8a, which is known as a regulator of LD fusion, were also decreased in the cells treated with NPPB. Other Cl- channel blockers, DIDS and IAA-94, also inhibited large LDs formation and Rab8a expression. These results demonstrate that Cl- channels do not regulate the adipocyte differentiation, but do regulate the LDs formation via Rab8a expression.Abbreviations: ASCs: adipose tissue-derived stem cells; LDs: lipid droplets; RUNX2: runt-related transcription factor 2; CFTR: cystic fibrosis transmembrane conductance regulator; TG: triacylglycerol; FA: fatty acid; GLUT4: glucose transporter type 4; ER: endoplasmic reticulum; ADRP: adipose differentiation-related protein; TIP47: tail-interacting protein of 47 kD; HSL: hormone sensitive lipase; PBS: phosphate-buffered saline; DMEM: Dulbecco's modified Eagle Medium; FBS: fetal bovine serum; SMA: smooth muscle actin; FAS: fatty acid synthase; ZONAB: ZO-1 associated nucleic acid binding protein; PPAR-γ: peroxisome proliferator-activated receptor-γ; C/EBPα: CCAAT/enhancer binding protein α; CE: cholesteryl ester; V-ATPase: vacuolar H+ ATPase.

Functional characterization of various channel-expressing central airway epithelial cells from mouse induced pluripotent stem cells.

Functional central airway epithelial cells (CAECs) from induced pluripotent stem cells (iPSCs) are an attractive potential cell source for central airway regeneration. The central airway epithelium, such as the tracheal epithelium, is composed of ciliated cells, goblet cells, and basal cells and has physiologically important functions such as the regulation of water volume on the airway surface by Cl- and water channels and the elimination of particles inhaled from the external environment by ciliary movement. Previous work from our group and from other research groups has reported the generation of airway epithelial cells from iPSCs. However, it remains unclear whether iPSC-derived CAECs express the various channels that are required for the regulation of water volume on the airway surface and whether these channels function properly. In this study, we generated CAECs from iPSCs supplemented with activin and bFGF using air-liquid interface culture. We then evaluated the physiological functioning of the iPSC-derived CAECs by examining the gene expression and transport functions of Cl - channels using a halide ion-sensitive yellow fluorescent protein and ciliary movement. Reverse-transcription polymerase chain reaction and immunohistochemistry indicated that various channel markers such as cystic fibrosis transmembrane conductance regulator (CFTR) and aquaporin (AQP) were present in iPSC-derived CAECs. Furthermore, the transport functions of Cl - channels and CFTR were successfully confirmed. Finally, ciliary movement was measured, and a ciliary beating frequency (CBF) of approximately 10 Hz was observed. These results demonstrate that CAECs generated by our method have physiological functions similar to those of native CAECs.

Carrageenan delays cell cycle progression in human cancer cells in vitro demonstrated by FUCCI imaging.

BACKGROUND: Carrageenan is a sulfated polysaccharide that exists in red seaweeds recently shown to have anticancer properties. Previous findings show various effects of carrageenan suppressing tumor cell growth. One of the hallmarks of cancer is uncontrolled proliferation, a consequence of loss of normal cell-cycle control, that underlies tumor growth. Recently there is an increasing interest in potential anticancer agents that affect cell cycle in cancer cells. Thus, in this study we investigated the effects of carrageenan on the tumor cell cycle. METHODS: Using human cervical carcinoma cells (HeLa) cells as and human umbilical vein endothelial cells (HUVEC), the cytotoxic effects of kappa carrageenan (k-CO) and lambda carrageenan (λ-CO) at the concentrations of 250-2500 µg/mL were observed. Cell viability was determined using the MTT assay while cell death rates were determined using staining with calcein-AM/propidium iodide. Cell-cycle profile and progression were demonstrated with HeLa cells expressing FUCCI (fluorescence ubiquitination-based cell-cycle indicator) probes (HeLa-FUCCI). RESULTS: Carrageenan had no significant effect on HUVEC (normal cells). In contrast both forms of carrageenan were cytotoxic towards HeLa cells (cancer cells). Furthermore, according to cell-cycle analysis with FUCCI cells, the cell cycle of HeLa cells was delayed in specific phases due to different carrageenan treatments. CONCLUSION: Considering these results, it could be suggested that carrageenan affects the cell-cycle of HeLa cells not only by arresting the cell cycle in specific phases but also by delaying the time needed for the cell to progress through the cell cycle. Additionally, different types of carrageenans have different effects on cell cycle progression. This effect of carrageenan towards cancer cells could possibly be developed into a tumor cell-specific anticancer agent.

Povidone-iodine-induced cell death in cultured human epithelial HeLa cells and rat oral mucosal tissue.

Although povidone-iodine (PVP-I) has been used as a gargle since 1956, its effectiveness and material safety have been remained controversial. The aim of this study was to investigate the toxicity of PVP-I to epithelial cells in a concentration range significantly lower than that used clinically. Study design was in vitro laboratory investigations and in vivo histological and immunologic analysis. We examined the effects of PVP-I at concentrations of 1 × 10(-2) to 1 × 10(3) µM and 1 × 10(-4) to 1 × 10 µM on HeLa cells as a model of epithelial cells and rat oral mucosa, respectively, after 1 or 2 days of exposure. Annexin V/FLUOS was used to distinguish live, apoptotic and necrotic cells. The terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) method was also used to observe whether apoptotic epithelial cells exist in rat oral mucosa after 1 day of exposure of PVP-I. HeLa cells developed concentration-dependent cytotoxicity, and epithelium of rat oral mucosa was thinned in a concentration-dependent manner. HeLa cell apoptosis increased after 1 × 10(0) µM of PVP-I exposure for 2 days. In the TUNEL method, many apoptotic epithelial cells were observed in the rat oral mucosa after 1 day of exposure to diluted 1 × 10(-2) µM of PVP-I, but minimal apoptotic epithelial cells were observed using 1 × 10(-3) µM of PVP-I. Our findings suggest that exposure to PVP-I, of which concentrations are even lower than those used clinically, causes toxicity in epithelial cells. This knowledge would help us better understand the risk of the use of PVP-I against mucosa.

Potential for respiratory epithelium regeneration from induced pluripotent stem cells.

OBJECTIVES: In cases of laryngeal inflammatory lesions and tracheal invasion of a malignant tumor, autologous tissue implantation techniques using skin or cartilage are often applied. However, these techniques are both invasive and unstable. The purpose of this study was to evaluate the potential use of induced pluripotent stem (iPS) cells in the regeneration of respiratory epithelium. METHODS: We seeded iPS cells on low-attachment plates in serum-free media to generate embryoid bodies (EBs). After a 3-day culture, the EBs were transferred to a gelatin-coated dish supplemented with activin A alone or with basic fibroblast growth factor (induction groups). As a control, EBs were cultured without these growth factors (control group). Cultured tissues from all groups were histologically examined for 2 weeks. RESULTS: In the induction groups, the presence of respiratory epithelium-like tissue was observed with hematoxylin and eosin staining after 14 days of culture. CONCLUSIONS: This study demonstrated the potential use of iPS cells in regeneration of the respiratory epithelium.

Reducing radiation exposure using commonly available objects.

OBJECTIVES: One and a half years have passed since the Fukushima Daiichi nuclear power plant disaster. The environmental radiation dose rate was not critical, but an existing exposure situation has been identified in a large part of Fukushima Prefecture. Although people continue to live and work in the contaminated area, they are not provided with sufficient information to reduce their exposure to radiation by themselves. In this study, we attempt to evaluate the efficiency of radiation shielding by using everyday items widely available to people. METHODS: NaI scintillation and Geiger-Müller survey meters were used to measure the radiation dose of (1) contaminated soil and (2) soil covered with commonly available items. RESULTS: In the soil at a depth of 10 cm from the surface, the radiation dose rate decreased from 3.36 to 0.65 µSv/h, and the count rate decreased from 3,120 to 352 cpm. Both the radiation dose rate and count rate reduced when the soil was covered with everyday items, such as a magazine more than 20 mm thick, a polystyrene foam board, and a wooden board of the same thickness. CONCLUSIONS: To protect residents from unnecessary radiation exposure in the existing exposure situation, covering contaminated soil with a wooden board or a magazine, either of them 20 mm thick, is useful to reduce the radiation dose.

Epitaxial growth of three-dimensionally architectured optoelectronic devices.

Optoelectronic devices have long benefited from structuring in multiple dimensions on microscopic length scales. However, preserving crystal epitaxy, a general necessity for good optoelectronic properties, while imparting a complex three-dimensional structure remains a significant challenge. Three-dimensional (3D) photonic crystals are one class of materials where epitaxy of 3D structures would enable new functionalities. Many 3D photonic crystal devices have been proposed, including zero-threshold lasers, low-loss waveguides, high-efficiency light-emitting diodes (LEDs) and solar cells, but have generally not been realized because of material limitations. Exciting concepts in metamaterials, including negative refraction and cloaking, could be made practical using 3D structures that incorporate electrically pumped gain elements to balance the inherent optical loss of such devices. Here we demonstrate the 3D-template-directed epitaxy of group III-V materials, which enables formation of 3D structured optoelectronic devices. We illustrate the power of this technique by fabricating an electrically driven 3D photonic crystal LED.

An assessment of radiation doses at an educational institution 57.8 km away from the Fukushima Daiichi nuclear power plant 1 month after the nuclear accident.

OBJECTIVES: On 11 March 2011, the Great East Japan Earthquake occurred. Due to this earthquake and subsequent tsunami, malfunctions occurred at the Fukushima Daiichi nuclear power plant. Radioactive material even reached the investigated educational institution despite being 57.8 km away from the power station. With the goal of ensuring the safety of our students, we decided to carry out a risk assessment of the premises of this educational institution by measuring radiation doses at certain locations, making it possible to calculate estimated radiation accumulation. METHODS: Systematic sampling was carried out at measurement points spaced at regular intervals for a total of 24 indoor and outdoor areas, with 137 measurements at heights of 1 cm and 100 cm above the ground surface. Radiation survey meters were used to measure environmental radiation doses. RESULTS: Radiation dose rates and count rates were higher outdoors than indoors, and higher 1 cm above the ground surface than at 100 cm. Radiation doses 1 cm above the ground surface were higher on grass and moss than on asphalt and soil. The estimated radiation exposure for a student spending an average of 11 h on site at this educational institution was 9.80 µSv. CONCLUSIONS: Environmental radiation doses at our educational institution 57.8 km away from the Fukushima Daiichi nuclear power plant 1 month after the accident were lower than the national regulation dose for schools (3.8 µSv/h) at most points. Differences in radiation doses depending on outdoor surface properties are important to note for risk reduction.

First detection and characterization of rat hepatitis E Virus (HEV-C1) in Japan.

Rat hepatitis E virus (HEV-C1) in the Orthohepevirus C species has been reported to cause zoonotic infection and hepatitis in humans. HEV-C1 strains have been detected from wild rats in many countries in Europe, Asia, and North America. However, in Japan, no HEV-C1 strains have been identified. In the present study, 5 (1.2%) of 428 wild rats (Rattus norvegicus or R. rattus) were positive for anti-HEV-C1 IgG. Although all 428 rat sera were negative for HEV-C1 RNA, it was detectable in 20 (19.8%) of 101 rat fecal samples collected on a swine farm, where HEV (genotype 3b, HEV-3b) was prevalent and wild rats were present. In addition, HEV-C1 RNA was detectable in the intestinal contents and liver tissues of 7 (18.9%) of 37 additional rats captured on the same farm. The HEV-C1 strain (ratEJM1703495L) obtained in this study shared only 75.8-84.7% identity with reported HEV-C1 strains over the entire genome but propagated efficiently in cultured cells. HEV-3b strains were detected in the rats' intestinal contents, with 97.3-99.5% identity to those in pigs on the same farm, but were undetectable in rat liver tissues, suggesting that wild rats do not support the replication of HEV-3b of swine origin.

The effect of topical amiloride eye drops on tear quantity in rabbits.

PURPOSE: To investigate the presence of epithelial sodium channels (ENaC) in rabbit and human conjunctival epithelium and to test the effects of topical amiloride, a potassium-sparing diuretic that blocks the ENaC, on tear quantity in rabbits. METHODS: Both healthy normal human and rabbit conjunctival tissues underwent immunohistochemistry staining for ENaC-α and γ subunits as well as for reverse transcription-polymerase chain reaction (RT-PCR) for detection of ENaC-α and ENaC-γ subunit mRNA expression. Rabbits were instilled topical amiloride eye drops and tear function tests were performed before and after instillations. RESULTS: Immunohistochemical staining for ENaC-α subunit in all rabbit eyes showed positive staining in apical and basal conjunctival epithelial cells. Human conjunctival epithelia revealed positive staining with ENaC-α antibody especially in the apical and basal layers. Immunohistochemistry staining with ENaC- γ antibody also revealed positive staining of the conjunctival epithelial cells especially in the basal layers. The ENaC-α mRNA was detected in samples from healthy white rabbit conjunctival epithelia, and ENaC-α and ENaC- γ mRNAs were detected in samples from healthy human conjunctival epithelia. The mean tear quantity showed a significant increase at 15 and 30 min compared to the pre-instillation value in eyes assigned to amiloride eye drops. The mean tear quantity at 15 and 30 min was significantly higher in the amiloride group compared to the control eyes. CONCLUSIONS: Topical amiloride application appears to increase the quantity of preocular tears owing to inhibition of conjunctival epithelial sodium channels.

Potential of induced pluripotent stem cells for the regeneration of the tracheal wall.

OBJECTIVES: Our previous studies focused on basic research and the clinical applications of an artificial trachea. However, the prefabricated artificial trachea cannot be utilized for pediatric airways, because the tracheal frame needs to expand as the child develops. The purpose of this study was to evaluate the potential of induced pluripotent stem (iPS) cells for the regeneration of the tracheal wall. METHODS: We cultured iPS cells in a 3-dimensional (3-D) scaffold in chondrocyte differentiation medium (bioengineered scaffold model), and the results were compared with those in a 3-D scaffold without iPS cells (control scaffold model). The 3-D scaffolds were implanted into tracheal defects in 8 nude rats. After 4 weeks, the regenerated tissue was histologically examined. RESULTS: Implanted iPS cells were confirmed to exist in all 5 rats implanted with bioengineered scaffolds. Cartilage-like tissue was observed in the regenerated tracheal wall in 2 of the 5 rats in the bioengineered scaffold model, but in none of the 3 rats in the control scaffold model. CONCLUSIONS: Implanted iPS cells were confirmed to exist in the bioengineered scaffold. Cartilage-like tissue was regenerated in the tracheal defect. This study demonstrated the potential of iPS cells in the regeneration of the tracheal wall.

Bioengineered trachea with fibroblasts in a rabbit model.

OBJECTIVES: Although our group has had mostly successful results with clinical application of a tracheal prosthesis, delayed epithelial regeneration remains a problem. In our previous studies using rats, it was demonstrated that tracheal fibroblasts accelerated proliferation and differentiation of the tracheal epithelium in vitro and in vivo. The purpose of this study was to evaluate the effects of fibroblasts on epithelial regeneration in larger tracheal defects in rabbits. METHODS: We developed a bioengineered scaffold, the luminal surface of which was coated with fibroblasts. This scaffold was implanted into tracheal defects in 12 rabbits (bioengineered group), and scaffolds without fibroblasts were implanted in 12 rabbits (control group). The regenerated epithelium was histologically examined by light microscopy, scanning electron microscopy, and immunohistochemical studies. RESULTS: In the bioengineered group, a stratified squamous epithelium was observed on the surface 7 days after transplantation. However, in the control group, the scaffolds were exposed. Fourteen days after implantation, a columnar ciliated epithelium was observed in the bioengineered group. The average thickness of the regenerated epithelium in the bioengineered group was significantly greater than that in the control group. CONCLUSIONS: This study indicated that fibroblasts had a stimulatory effect that hastened regeneration of the epithelium in large tracheal defects.

Formation of a photocatalytic WO3 surface layer on electrodeposited Al-W alloy coatings by selective dissolution and heat treatment.

In this study, we explored the feasibility of WO3 surface layer formation on electrodeposited Al-W alloy coatings by selective dissolution and heat treatment, with the aim of providing corrosion-resistant Al-W alloy coatings with photocatalytic self-cleaning properties under visible light illumination. The selective dissolution of Al and oxidation of residual W was carried out by immersing Al-W alloy films in an aqueous solution of nitric acid. A nanostructured H2WO4·H2O surface layer was formed on the alloy film by this process. The H2WO4·H2O layer was dehydrated to WO3 by heat treatment, yielding a multilayered WO3/Al-W alloy film with an approximately 300 nm thick WO3 layer. The WO3/Al-W alloy film exhibited photocatalytic self-cleaning, as demonstrated by the photodegradation of stearic acid and methylene blue. We also confirmed that selective dissolution and heat treatment did not significantly diminish the corrosion resistance of the Al-W alloy films.

The Cytotoxic Effects of Geranylgeranylacetone Are Attenuated in the High-Glucose Condition.

Geranylgeranylacetone (GGA) has been used as an antiulcer drug and also is known as inducer of heat shock protein 70 that has cytoprotective effects especially in hyperglycemic condition. In contrast, cytotoxicity of GGA has also been reported. Some studies have reported that GGA suppresses cell growth and induces apoptosis in cell models of human leukemia, ovarian carcinoma, and colon cancer in vitro. Therefore, the aim of this study was to determine whether GGA can have a cytotoxic effect on a human cervical cancer cell line (HeLa), human colorectal adenocarcinoma cells (Caco-2), and human embryonic kidney cells 293 (HEK) in normal-glucose and high-glucose environments (NG and HG, respectively). The results showed that 100 µM GGA inhibited proliferation of HeLa cells only in NG environment despite inhibiting proliferation of Caco-2 and HEK cells regardless of glucose concentration. Cell viability assay revealed that GGA decreased viability of HeLa, Caco-2, and HEK cells only in NG environment. Flow cytometric analyses revealed that the type of cell death was a combination of necrosis and apoptosis. Our study revealed that difference in cytotoxicity of GGA is influenced by glucose condition. The cytotoxic effects of GGA are attenuated in the HG condition. Since both cytotoxic and cytoprotective effects are reported about GGA, further research is needed about the mechanism of the cytotoxic effects.

UVA Photoprotective Activity of Brown Macroalgae Sargassum cristafolium.

Sunscreens today contain several synthetic UV (Ultraviolet) filter molecules to protect the skin epidermis from UV radiation damage. However, these molecules may create several negative effects on human skin. Due to this condition, there is an increase in the development of natural products to replace uses of these synthetic chemicals. Brown macroalgae Sargassum has been recently studied for its photoprotective activities. The purpose of this study is to investigate photoprotective activity of one of most abundant Sargassum species in Lombok coast; Sargassum cristaefolium. Spectrophotometry analysis with UV-VIS revealed the UV spectra absorbing capability of Sargassum cristaefolium (SC) in the UVA spectrum range (314-400 nm). Furthermore, spectrometry analyses with LC-MS revealed the existence of UV absorbing compound MAA-palythene. In correlation, SC ethanol extracts also demonstrate that it could protect DNA from UVA irradiation as analyzed in vitro in HeLa cell model. The effects of SC on UVA exposed-dorsal mice skin have also shown interesting results, as mice pretreated with SC before UVA exposure showed protective activity on the epidermal integrity similar as positive control. Whereas, UV exposed mice without SC or commercial products resulted in increased epidermal thickness, which is the common parameter of skin photoaging. In addition, pretreated mice with SC also show protective effects in the formation of collagen connective tissues. Overall, current results show promising photoprotective activity of SC against UV radiation. More advanced investigations of SC as a potential photoprotective agent would be reasonable for development of macroalgae-based natural skin protection products.

Potential of heterotopic fibroblasts as autologous transplanted cells for tracheal epithelial regeneration.

The tracheal epithelium maintains the health of the respiratory tract through mucociliary clearance and regulation of ion and water balance. When the trachea is surgically removed, artificial grafts have been clinically used by our group to regenerate the trachea. In such cases, the tracheal epithelium needs 2 months for functional regeneration. Previous study has shown that fibroblasts facilitate tracheal epithelial regeneration. In this study, heterotopic fibroblasts originating from the dermis, nasal, and gingival mucosa were cocultured with tracheal epithelial cells to evaluate their potential as autologous transplanted cells for tracheal epithelial regeneration. The epithelia induced by the heterotopic fibroblasts showed differences in structure, cilia development, mucin secretion, and expression of ion and water channels. These results indicated that nasal fibroblasts could not induce mature tracheal epithelium and that dermal fibroblasts induced epidermis-like epithelium. Only the gingival fibroblasts (GFBs) could induce morphologically and functionally normalized tracheal epithelium comparable to the epithelium induced by tracheal fibroblasts. Epithelial cell proliferation and migration were also upregulated by GFBs. These results indicate that GFBs are useful as autologous transplant cells for tracheal epithelial regeneration.

Full-length genome of a novel genotype 3 hepatitis E virus strain obtained from domestic pigs in Japan.

Hepatitis E virus (HEV) causes acute or chronic hepatitis in humans and can be transmitted via the fecal-oral route. Pigs are one of the main reservoirs for this infection. Sixty pigs, 4-5 months of age, on a swine herd in Japan had detectable anti-HEV IgG antibodies, and five (8.3%) of them had ongoing infection of genotype 3 HEV. Five HEV strains obtained from the viremic pigs shared 98.8-100% nucleotide identity, and one representative strain (swHE1606845), whose entire genomic sequence was determined in this study, differed by 14.1-19.6% from the reported HEV strains of subtypes 3a-3k and by 14.7-19.1% from other genotype 3 HEV strains whose subtypes have not yet been assigned. swHE1606845 showed a higher nucleotide p-distance value of ≥0.143 with the genotype 3 HEV strains of subtypes 3a-3k and ≥0.152 with other genotype 3 strains of unassigned subtypes. A SimPlot analysis revealed a lack of recombination events. These results indicate that swHE1606845 is a candidate member of a novel subtype of genotype 3. Further efforts to identify the swHE1606845-like novel strain are warranted to clarify the origin of this strain and to determine the complete nucleotide sequences of two additional swHE1606845-like strains for assigning a new subtype.

Effect of fibroblasts on tracheal epithelial regeneration in vitro.

Several artificial grafts for covering deficient trachea have been produced through tissue engineering. Recently, our group clinically used an artificial trachea made from collagen sponge for patients with noncircumferential tracheal resection. However, the slowness of epithelial regeneration on the surface of the artificial trachea was confirmed as one particular problem. In this study, we co-cultured tracheal epithelial cells with fibroblasts and examined effects of fibroblasts on epithelial regeneration in vitro. Fibroblasts activated epithelial cell proliferation and migration. In co-culture with fibroblasts, epithelial cells reconstructed pseudostratified epithelium, which was composed of ciliated, goblet, and basal cells. Furthermore, a basement membrane was reconstructed between epithelial cells and fibroblasts, and integrin beta4 was also observed there. Fibroblasts rapidly increased mucin secretion by epithelial cells. These results indicate that stimulatory effects of fibroblasts on epithelial cell migration, proliferation, and differentiation would reduce the time required for covering of epithelial cells on the defect of luminal surface and hasten regeneration of morphologically and functionally normalized epithelium involving the reconstruction of basement membrane.

Mild hypertension in young Kurosawa and Kusanagi-hypercholesterolaemic (KHC) rabbits.

The coexistence of hypertension and hypercholesterolaemia from youth may increase the prevalence of and mortality from cardiovascular disease and stroke. We thus investigated haemodynamics of mild hypertension in young Kurosawa and Kusanagi-hypercholesterolaemic (KHC) rabbits aged 10-12 months old, as models of heritable hypercholesterolaemia. Pressure and flow waves were simultaneously recorded at the ascending aorta with a catheter-tip micromanometer and ultrasonic flow meter under pentobarbital anaesthesia, respectively. Systolic (119.3 +/- 6.5 and 138.4 +/- 7.4 mmHg (mean +/- SD) for control and KHC rabbit groups; p < 0.001), diastolic (95.7 +/- 6.1 and 109.8 +/- 5.2; p < 0.001), mean (105.8 +/- 6.5 and 122.5 +/- 4.9; p < 0.001) and pulse (23.7 +/- 2.5 and 28.6 +/- 4.0; p < 0.001) pressures as well as total peripheral vascular resistance (0.32 +/- 0.02 and 0.37 +/- 0.03 mmHg/ml/min; p < 0.001) were significantly greater in the KHC rabbit group than those in the age-matched control rabbit group, respectively, while there were no significant differences in the mean aortic flow, heart rate or stroke volume between the two rabbit groups. Aortic input impedance (p < 0.05) and reflection coefficient (p < 0.05) were significantly greater at lower frequency in the KHC rabbit group than in the control rabbit group, whereas there was no significant difference in the characteristic impedance between the two rabbit groups. Plasma angiotensin I (p < 0.01) and II (p < 0.01) levels and serum angiotensin converting enzyme activity (p < 0.05) were significantly greater in the KHC rabbit group than in the age-matched control rabbit group. Atheromatous plaque was in the early stage and composed mainly of abundant foam cells. Neither sclerotic lesions nor stenosis were observed in main peripheral arteries. The mild hypertension in young KHC rabbits was due partly to the increased activity of the renin-angiotensin system. These findings may be thought provoking in elucidating the mechanism and developing preventive and therapeutic strategies in young patients with coexistent hypertension and hypercholesterolaemia.

Tissue engineering for regeneration of the tracheal epithelium.

OBJECTIVES: The slowness of epithelialization on the artificial trachea that has been successfully used in humans is a problem. The purpose of this study was to develop a way to regenerate the epithelium on the surface of this artificial trachea. METHODS: In an in vitro study, isolated rat tracheal epithelial cells were seeded on a collagenous gel that was stratified on a collagenous sponge. Histologic and immunohistochemical examinations were made. In an in vivo study, we transplanted grafts with green fluorescent protein-positive tracheal epithelial cells onto the tracheal defects of normal rats. At 3, 7, 14, and 30 days after the operation, histologic and immunohistochemical examinations were made. RESULTS: In the in vitro study, the 3 layers--the epithelium, gel, and sponge--could be observed. The epithelium expressed cytokeratin 14, cytokeratin 18, and occludin. In the in vivo study, the artificial trachea was covered with epithelium at 3 days after operation, and then the epithelium differentiated from single- or double-stratified squamous epithelium into columnar ciliated epithelium. Green fluorescent protein-positive cells were found 3 days after operation. CONCLUSIONS: We believe that the method used in our experiment is an effective way to regenerate the epithelium on the surface of an artificial trachea. With further experimentation, this method should be suitable for clinical application.