Keratinocyte Growth Factor Gene Electroporation into Skeletal Muscle as a Novel Gene Therapeutic Approach for Elastase-Induced Pulmonary Emphysema in Mice

Pulmonary emphysema is a progressive disease with airspace destruction and an effective therapy is needed. Keratinocyte growth factor (KGF) promotes pulmonary epithelial proliferation and has the potential to induce lung regeneration. The aim of this study was to determine the possibility of using KGF gene therapy for treatment of a mouse emphysema model induced by porcine pancreatic elastase (PPE). Eight-week-old BALB/c male mice treated with intra-tracheal PPE administration were transfected with 80 μg of a recombinant human KGF (rhKGF)-expressing FLAG-CMV14 plasmid (pKGF-FLAG gene), or with the pFLAG gene expressing plasmid as a control, into the quadriceps muscle by electroporation. In the lung, the expression of proliferating cell nuclear antigen (PCNA) was augmented, and surfactant protein A (SP-A) and KGF receptor (KGFR) were co-expressed in PCNA-positive cells. Moreover, endogenous KGF and KGFR gene expression increased significantly by pKGF-FLAG gene transfection. Arterial blood gas analysis revealed that the PaO2 level was not significantly reduced on day 14 after PPE instillation with pKGF-FLAG gene transfection compared to that of normal mice. These results indicated that KGF gene therapy with electroporation stimulated lung epithelial proliferation and protected depression of pulmonary function in a mouse emphysema model, suggesting a possible method of treating pulmonary emphysema

Intratracheal Administration of Recombinant Human Keratinocyte Growth Factor Promotes Alveolar Epithelial Cell Proliferation during Compensatory Lung Growth in Rat

Keratinocyte growth factor (KGF) is considered to be one of the most important mitogens for lung epithelial cells. The objectives of this study were to confirm the effectiveness of intratracheal injection of recombinant human KGF (rhKGF) during compensatory lung growth and to optimize the instillation protocol. Here, trilobectomy in adult rat was performed, followed by intratracheal rhKGF instillation with low (0.4 mg/kg) and high (4 mg/kg) doses at various time-points. The proliferation of alveolar cells was assessed by the immunostaining for proliferating cell nuclear antigen (PCNA) in the residual lung. We also investigated other immunohistochemical parameters such as KGF, KGF receptor and surfactant protein A as well as terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling. Consequently, intratracheal single injection of rhKGF in high dose group significantly increased PCNA labeling index (LI) of alveolar cells in the remaining lung. Surprisingly, there was no difference in PCNA LI between low and high doses of rhKGF with daily injection, and PCNA LI reached a plateau level with 2 days-consecutive administration (about 60%). Our results indicate that even at low dose, daily intratracheal injection is effective to maintain high proliferative states during the early phase of compensatory lung growth

Keratinocyte growth factor accelerates compensatory growth in the remaining lung after trilobectomy in rats.

OBJECTIVE: In rats pulmonary resection is followed by lung compensatory growth. However, the molecular mechanism underlying lung compensatory growth remains unclear. Keratinocyte growth factor is expressed in lung tissue and is considered a possible mitogen for lung epithelial cells. The objectives of this study were to define the role of keratinocyte growth factor and its receptor in rat lung compensatory growth after trilobectomy and the effect of exogenous keratinocyte growth factor gene transfection. METHODS: Adult Lewis rats were used. Right trilobectomy was performed in the operation group and sham thoracotomy in the sham group. In the operation group, keratinocyte growth factor-FLAG or FLAG expression vector was transfected directly into the lung by means of electroporation. Expression of keratinocyte growth factor and its receptor and alveolar cell proliferation index based on proliferating cell nuclear antigen levels were measured in the right lung at day 14 after the operation. RESULTS: Proliferating cell nuclear antigen, keratinocyte growth factor, and keratinocyte growth factor receptor expression in lung epithelial cells was significantly increased at day 4 after trilobectomy. Transfection of keratinocyte growth factor-FLAG expression vector resulted in further significant enhancement of proliferating cell nuclear antigen at day 4 after trilobectomy; however, the transfection of FLAG expression vector did not alter the enhancement of proliferating cell nuclear antigen. Exogenous expression of keratinocyte growth factor in the remaining lung by means of electroporation significantly augmented epithelial proliferation and decreased the average airspace distance (mean linear intercept). CONCLUSION: Our results implicate keratinocyte growth factor in the induction of alveolar epithelial cell proliferation for compensatory lung growth and indicate that overexpression of keratinocyte growth factor in the remaining lung by means of electroporation significantly augmented lung epithelial proliferation

Prediction of portal pressure from intraoperative ultrasonography

BackgroundPortal hypertension is a major risk factor for hepatic failure or bleeding in patients who have undergone hepatectomy, but it cannot be measured indirectly. We attempted to evaluate the intraoperative ultrasonography parameters that correlate with portal pressure (PP) in patients undergoing hepatectomy.MethodsWe examined 30 patients in whom PP was directly measured during surgery. The background liver conditions included chronic viral liver disease in seven patients, chemotherapy-associated steatohepatitis in four patients, fatty liver in one patient, hepatolithiasis in one patient, obstructive jaundice in one patient, and a normal liver in 16 patients. A multivariate logistic analysis and linear regression analysis were conducted to develop a predictive formula for PP.ResultsThe mean PP was 10.4 ± 4.1 mm Hg. The PP tended to be increased in patients with chronic viral hepatitis. A univariate analysis identified the association of the six following parameters with PP: the platelet count and the maximum (max), minimum (min), endo-diastolic, peak-systolic, and mean velocity in the portal vein (PV) flow. Using multiple linear regression analysis, the predictive formula using the PV max and min was as follows: Y (estimated PP) = 18.235?0.120 × (PV max.[m/s])?0.364 × (PV min). The calculated PP (10.44 ± 2.61 mm Hg) was nearly the same as the actual PP (10.43 ± 4.07 mm Hg). However, there was no significant relationship between the calculated PP and the intraoperative blood loss and post hepatectomy morbidity.ConclusionsThis formula, which uses ultrasonographic Doppler flow parameters, appears to be useful for predicting PP

Accuracy and prognostic impact of a vessel invasion grading system for stage IA non-small cell lung cancer.

BACKGROUND: Despite the recognition that lymphatic and blood vessel invasion is an important prognostic factor in lung cancer, there is no common definition for pathological evaluation of vessel invasion. The aim of the present study was to determine whether D2-40 immunostaining can increase the accuracy of detection of lymphatic vessel invasion and whether our new grading system of vessel invasion by "degree" could be used instead of conventional evaluation by "presence" for pathological stage IA non-small cell lung cancer (NSCLC). METHODS: The vessel invasion classification was re-evaluated in 221 recent paraffin-embedded sections of p-stage IA NSCLC stained by Hematoxylin-Eosin (HE), Elastica-Van-Gieson (EVG), and D2-40. RESULTS: After re-assessment using D2-40 immunostaining, 41.2% (31 of 75) of ly1 cases by HE/EVG changed to ly0, and 14.9% (17 of 114) of ly0 cases by HE/EVG changed to ly1. Overall, 4 of 28 ly2 cases on conventional staining were changed to ly1, and 2 were changed to ly0 using D2-40 immunostaining. When the patients were divided into two groups by the presence of vessel invasion (v/ly0 vs. 1, 2, 3), there was no significant difference in cancer-specific survival (p=0.1107, 0.0875, respectively), while when they were divided according to degree of vessel invasion (v/ly0, 1 vs. 2, 3), there was a statistically significant difference (p=0.0038, p=0.0002, respectively). On multivariate analysis, lymphatic vessel invasion had a significant impact on cancer-specific survival (p=0.0061). CONCLUSION: Our results suggest that D2-40 immunostaining provides a precise diagnosis of lymphatic vessel invasion, and our new grading system of vessel invasion by "degree" is accurate and has prognostic value in early lung cancer

Acoustic injuries due to headphones among high school students

A survey was performed at 3 junior and 2 senior high schools in Kagawa Prefecture, in order to carify the use of headphones and the possibility of auditory damage due to headphones. Out of 2,217 students, 46.1% used “headphones”, including earphones and so-called Walkman-type headphones. Most of them used the headphones to listen to music. There was a significant difference in the incidence of cochlear symptoms between headphones users and non-users. In junior high school students, the incidence of cochlear symptoms was higher in those who had used headphones for more than four years and in those who used headphones for more than two hours per day. The extent of use correlated with the incidence of cochlear symptoms and the results indicate the possibility that acoustic injury could be caused by headphones

Discussion on regional revitalization using woody biomass resources as renewable energy

Expanding the use of renewable energy is a matter of concern in many countries. Many Japanese local municipalities are attempting to promote business creation using renewable energy as an opportunity for regional revitalization based on the policy of the Japanese government. Renewable energy makes significant contributions to the region in terms of energy security, stable energy supply, and job creation. However, the most important contribution is enhancing residents' attachment to their community and supporting actions that appeal to the community through renewable energy utilization. Residents should actively participate in regional revitalization, which leads to sustainable prosperity of the region. However, the current measures of local municipalities seem to be lacking such a viewpoint. This study aims to analyse the environmental and economic aspects as well as the social acceptance of establishing a bathing business that utilizes woody biomass as its heat source. This study also discussed how the generation of heat from woody biomass contributes to regional revitalization. The case study area was Kobe and Mt. Rokko, Japan. First, heat production and utilization was assumed for the forest resources in Mt. Rokko, and the annual biomass energy production potential was estimated. The annual costs of lumber cutting, transportation, and chipping, and the annual CO2 reduction by substituting bunker A with woody biomass for heat production were also estimated. A questionnaire survey was conducted to determine community awareness of energy production using woody biomass from Mt. Rokko. Principal component analysis, contingent valuation method, and path analysis were used to quantify respondents' awareness

Field survey of acatalasemia and hypocatalasemia from 1949 through 1985

The authors reported the results of a field survey of acatalasemia and hypocatalasemia conducted from 1949 through 1985. Six families and eleven cases of acatalasemia and twelve hypocatalasemia cases were added to the statistics reported in 1976. As a result, the total number of reported acatalasemias became 108 cases in 53 families as of the end of 1985