The shortage of medical doctors is now a serious social problem in Japan. There are 2.1 physicians per 1000 residents in Japan, fewer than most other OECD countries. Tokushima Prefecture has the second highest population of medical doctors in Japan, but the shortage of medical doctors in rural areas was observed by an uneven distribution. Primary care practice was started in the education of clinical clerkship for 5^th grade medical students of our university since July, 2008. They round a variety of hospitals, clinics and facilities for nursing-care during one week in Kaihu county where has a severe problem in the shortage of medical doctors in Tokushima Prefecture. In order to research the efficacy of the primary care practice, we have administered a questionnaire for medical students before and after the lecture and practice, and the intensity was estimated in each student by using visual analogue scale. The results showed that the practice was more meaningful in increasing the intensity for interest and passion for community medicine and medicine in remote area than the lecture. Our data indicate that it is important to prepare more courses to learn primary care and general medicine in our clinical practice system to continue the interest and passion in community medicine. J. Med. Invest. 56: 81-87, August, 2009

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Insulin resistance is one of the pathophysiological features of obesity and type 2 diabetes. Recent findings have linked insulin resistance to chronic low-grade inflammation in white adipose tissue. Excess storage of saturated fat in white adipose tissue due to a modern life style causes hypertrophy and hyperplasia of adipocytes, which exhibit attenuated insulin signaling due to their production and release of saturated fatty acids. These adipocytes recruit macrophages to white adipose tissue and, together with them, initiate a proinflammatory response. Proinflammatory factors and saturated fatty acids secreted into the bloodstream from white adipose tissue impair insulin signaling in non-adipose tissues, which causes whole-body insulin resistance. J. Med. Invest. 56: 88-92, August, 2009

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The Long-Evans Cinnamon (LEC) rat, an animal model of Wilson’s disease, is resistant to a variety of chemical carcinogenesis except liver and colon. In the present study, N-ethyl-N-nitrosourea (ENU)-induced transplacental carcinogenesis was examined in male and female LEC, Long-Evans Agouti (LEA), a sibling line of the LEC rat, and F344 rats (n=21). ENU was administered to pregnant rats as a single s.c. injection at a dose of 60 mg/kg body weight on the 17th day after conception. Cerebral/spinal gliomas and trigeminal/spinal nerve schwannomas developed in both LEA and F344 rats at 30 weeks of age, but no nervous system tumors developed in LEC rats, the difference being statistically significant. Lung adenomas also developed in LEA and F344 rats, but not in LEC rats. Semiquantitative RT-PCR demonstrated that metallothionein (MT)1a, MT2 and O⁶-methylguanine-DNA methyltransferase (MGMT) mRNA levels in the liver of LEC rats were higher than those in F344 and LEA rats. In addition, Western blot analysis showed that MT (MT1 plus MT2) in the liver of LEC rats was also higher than that in other strains. Present results suggest that high levels of MT and/or MGMT contribute to the resistance to nitrosamine-induced carcinogenesis in LEC rats. J. Med. Invest. 56: 93-98, August, 2009

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We evaluated risk factors of radiation pneumonitis (RP) after whole breast irradiation following breast-conserving surgery. Four hundred and seventy-two cases underwent whole breast irradiation with tangential field following breast-conserving surgery in our hospital, between January 2005 and April 2007. Of these cases, we performed statistical analyses for 423 breasts of 413 patients, using a pulmonary dose-volume histogram. Patient characteristics, treatment regimens and irradiation methods were included as variables in the analyses on risk factors of RP. As a result, 89 breasts of 84 cases (21%) were diagnosed with RP. The version 3.0 of the NCI Common Terminology Criteria for Adverse Events was used to evaluate the grade of pneumonitis: 77 cases (18.2%) were diagnosed as Grade 1 RP, 10 cases (2.3%) as Grade 2, and 2 cases (0.5%) as Grade 3. Multivariate analysis indicated that the significant risk factors for RP were central lung distance (CLD) (>1.8 cm) and the short axis length of the radiation field. The incidence of radiation-induced bronchiolitis obliterans organizing pneumonia (BOOP) syndrome significantly correlated only with CLD. The lung volume within the radiation field was shown to be a significant risk factor for RP and radiation-induced BOOP syndrome. J. Med. Invest. 56: 99-110, August, 2009

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Purpose: To report the efficacy of intravitreal injections of bevacizumab for diabetic macular edema (DME) in the short-term. Design: Retrospective, noncomparative, interventional case series. Methods: Medical records of 20 eyes of 19 patients who underwent intravitreal injections of bevacizumab for persistent diabetic macular edema were reviewed retrospectively. All eyes received intravitreal injections of bevacizumab (1.25 mg/0.05 ml). The clinical course of best-corrected visual acuity (BCVA) using a logarithm of the minimum angle of resolution chart, and averaged foveal retinal thickness using an optical coherence tomography (OCT) were monitored for up to four weeks after the injection. Results: BCVA at one week improved by two lines or more in six eyes (30%) and in nine eyes (45%) at four weeks. However, no significant improvement in the mean BCVA from baseline was observed at one week (P>0.05) and four weeks (P>0.05). Mean retinal thicknesses (RT) were 411±170 μm at baseline, 349±102 μm at one week after the injection (P<0.05), and 380±159 μm at four weeks (P>0.05). One week after the injection, significant regression of macular edema was seen. However, recurrence occurred at four weeks. No complications such as severe vision loss, endophthalmitis, or systemic events developed. Conclusion: No changes in BCVA and RT were observed in the short-term observation after the intravitreal injection of bevacizumab for DME. J. Med. Invest. 56: 111-115, August, 2009

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Purpose: To evaluate the efficacy of combined photodynamic therapy (PDT) and sub-Tenon’s capsule injection of triamcinolone acetonide (TA) for the treatment of choroidal neovascularization (CNV) in age-related macular degeneration (AMD) at 1-year follow-up. Methods: A total of 117 eyes with subfoveal CNV caused by AMD were included. Sixty-eight eyes were treated with combined PDT and sub-tenon injection of 20 mg TA and compared with a control group of 49 eyes treated with PDT alone. All patients completed 1 year follow-up. The main outcome measures were best-corrected visual acuity (BCVA) and retreatment frequency. Results: There were no significant differences in age, sex, VA, lesion type, and greatest linear dimension between the two groups at baseline. BCVA analysis showed statistically no significant differences between the two groups. The PDT+TA group required a lower mean number of treatments (1.38 versus 1.67, P=0.032). Conclusion: Our study confirms that sub-Tenon’s capsule injection of TA with PDT significantly reduces the retreatment frequency of PDT. J. Med. Invest. 56: 116-119, August, 2009

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Objectives: To clarify the underlying mechanisms of L-DOPA induced vasoconstriction in rat aorta. Methods: The effect of L-DOPA on phenylephrine-induced contractile force of blood vessels was examined in vitro using rat aortic ring preparations by isometric tension experiment. Involvement of nitric oxide (NO) in the effect of L-DOPA on vascular smooth muscle was studied by using N_ω-Nitro-L-arginine (L-NNA), Sodium nitroprusside (SNP) in endothelium-intact and endothelium-denuded aortic rings. Results: L-DOPA potentiated α-adrenergic receptor- and depolarization-induced vascular contraction and inhibited acetylcholine-induced vasorelaxation. This effect was diminished by pretreatment of the aortic rings with L-NNA, an inhibitor of NO synthesis, or by removing the endothelium from the ring preparations. In endothelium-denuded rings, L-DOPA inhibited exogenous NO-dependent but not cGMP-mediated vasorelaxation. Increases in cGMP levels in response to an NO donor were attenuated by L-DOPA in cultured rat aortic smooth muscle cells. L-DOPA could not contract rings (without endothelium) pretreated with 3-(5’-hydroxymethyl- 2’-furyl)-1-benzyl indazole (YC-1), an activator of guanylyl cyclase, but SOD (150 U/ml) pretreatment of rings with endothelium inhibited contraction by L-DOPA. Conclusions: These results suggest that L-DOPA inhibits nitric-dependent vasorelaxation on vascular smooth muscle cells via production of reactive oxygen species. J. Med. Invest. 56: 120-129, August, 2009

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Low calcium intake and physical inactivity are modifiable risk factors of osteoporosis; however, little information is available about the prevalence of these risk factors among urban Filipino women living on low-incomes. The present study, therefore, investigated daily calcium intake, main calcium sources, and physical activity status in this population. The study group comprised healthy women aged in their 30 s who had participated in our previous survey using heel speed of sound (SOS) measurement in Davao, Philippines. The women were stratified into three groups based on SOS score and 20 were randomly selected from each, giving 60 in total. Calcium intake was measured by direct analysis of the food samples collected from 3-days 24 hour-food duplicate method. Physical activity was estimated based on pedometer determined walking steps over 5 days. The median [25%, 75%] calcium intake per day was 289 [225, 434] mg. Traditional foods derived from local small fish and plants were the main calcium sources. The median walking steps per day was 8750 [6920, 10836]. Although three groups did not show significantly different calcium intakes and walking steps, consumption of low-cost small fish and plant foods could be encouraged along with walking in urban Filipino women living on low-incomes. J. Med. Invest. 56: 130-135, August, 2009

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Objective We report the findings of Thallium-201 chloride (²⁰¹TlCl) scintigraphy and consider how to use this technique to evaluate the character of soft tissue lesions. Patients and Methods We enrolled 91 consecutive patients (45 males and 46 females, age range 8-91-years-old). Nineteen patients were malignant and 72 were benign. Patients were scanned 15 minutes (early phase) and 3 hours (delayed phase) after ²⁰¹TlCl injection. More intense uptake in the lesion compared to the normal side was considered as ‘high’, the same degree of uptake was considered ‘iso’, and decreased uptake was ‘low’. The retention index (RI) was calculated in 9 patients in the malignant group and in 16 patients in the benign group. Results In malignant tumors, 15 of 19 patients showed high uptake in both the early and delayed phases. One malignant fibrous histiocytoma patient was high only in the delayed phase and 1 liposarcoma patient was high only in the early phase. Two liposarcoma patients showed an iso uptake in both phases. One of these patients was pathologically diagnosed as a myxoid type. In benign lesions, no lipoma showed increased uptake. All neurogenic tumors except for 2 demonstrated high uptake. All 3 ganglions of the lower extremities showed iso uptake. Most inflammatory diseases showed increased uptake. Clinically-considered benign patients consisted of tumorous lesions or inflammatory disease. Only 2 patients were considered ‘low’, and these were diagnosed as intramuscular hematoma and cyst. RI was variable in both malignant and benign lesions and no statistically significant difference was seen between malignant and benign lesions by t-test (p=0.72). Conclusions A high ²⁰¹TlCl uptake lesion is more frequently seen in malignant tumors, but regardless of whether the tumor is benign or malignant, according to the histopathological variety, the ²⁰¹TlCl uptake pattern can not be the only indicator to differentiate malignant from benign tumors. We ultimately need to evaluate the nature of tumors by a combination of several imaging techniques. J. Med. Invest. 56: 136-141, August, 2009

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Despite the popularity of 3T3-L1 adipocytes as a model system of adipocytes in vivo, they do not carry all of the cellular functions of adipocytes in vivo. In this study, we investigated the effect of extracellular matrix (ECM) rigidity on insulin signal transduction in 3T3-L1 adipocytes. On 250 Pa polyacrylamide gel (soft gel) laminated with a mixture of collagen type 1 and fibronectin, whose rigidity matches that of adipose tissue, expression of the insulin receptor, IRS-1 and AKT was upregulated and their insulin-stimulated phosphorylation was enhanced. Furthermore, the expression of GLUT1 was downregulated, whereas the expression of GLUT4 was unaffected as ECM rigidity decreased. Insulin-stimulated GLUT4 recruitment to the plasma membrane was significantly enhanced in cells seeded on soft gel. These results suggest that adjusting the ECM rigidity to that of adipose tissue augments insulin signaling in 3T3-L1 adipocytes and enhances insulin-stimulated GLUT4 recruitment to the plasma membrane. J. Med. Invest. 56: 142-149, August, 2009

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We previously developed a high powered light-emitting diode device capable of discharging germicidal ultraviolet irradiation (UVA-LED) at an approximate wavelength of 365 nm. This study examined the bactericidal activity of UVA-LED in moving air streams. Aerosols of Escherichia coli DH5α were exposed to UVA-LED irradiation using a stable current (0.5 A and 1.2 mW/cm²) or pulse current (1.0 A and 0.2 mW/cm²). Settle plate analysis was used for bioaerosol sampling, where results were expressed as Colony Forming Units. A -3 Log inactivation of the E. coli population occurred after 75 minutes of constant exposure to stable current. The pulse current produced inactivation within a similar timeframe. Our results might be significant as a basic study for further investigations about the effect of UVA-LED on airborne bacteria and its suitability for air disinfection applications. J. Med. Invest. 56: 150-156, August, 2009

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Previously, we noticed the presence of nerve fiber-like structures in a whole mount preparation of the rat cornea that had not been stained with the non-specific acetylcholinesterase (NsAchE) method. These nerve-like fibers were projected into the central area of the cornea, forming a mesh-like pattern. The aim of this study is to examine the properties of these mesh-like fibers using the following two methods: their sensitivity to capsaicin and the detection of isolectin B4 (IB4)- and capsaicin receptor TRPV1 (transient receptor potential vanilloid 1)-reactivities. The mean disappeared area of non-stained fibers after NsAchE treatment was 26% of the total areas in the neonatally capsaicin-treated cornea. Bunches composed of fine IB4-positive nerve fibers were seen in a whole mount preparation. There were connections between the bunches, producing a mesh-like pattern similar to that of the fibers that were not stained with NsAchE. Fine TRPV1-immunoreactive (ir) nerve fibers were also shown to form bunches, with connections between each bunch observed in whole mount preparations. Thus, TRPV1-ir nerve fibers seem to densely innervate the rat corneal subepithelial stroma and are distinct from the NsAchE-positive nerve fibers. The TRPV1-ir fine nerve fibers overlapped with the IB4-positive nerve fibers, suggesting that the mesh-like fibers that were not stained with NsAchE are fine nociceptive sensory nerve fibers because of their sensitivity to capsaicin and similar distribution pattern to IB4- and TRPV1-positive nerve fibers. J. Med. Invest. 56: 157-165, August, 2009

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Acanthamoeba keratitis (AK) is a severe infectious corneal ulcer that usually occurs in contact lens wearers. Although the number of AK cases in Japan has been increasing, many of these cases are diagnosed in the early stage and are treated adequately. This is probably because of the increased availability of various diagnostic techniques and the ever-increasing knowledge about AK among ophthalmologists. In this article, we described 3 cases of AK that were diagnosed and treated in the early stages of the disease, and we discuss why 1 of the cases had a less favorable prognosis than the other 2 cases, which had excellent prognoses, from an etiological point of view. J. Med. Invest. 56: 166-169, August, 2009

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