Development of a high-sensitivity and portable cell using Helmholtz resonance for noninvasive blood glucose-level measurement based on photoacoustic spectroscopy.

We describe the possibility of high-sensitivity noninvasive blood glucose measurement based on photoacoustic spectroscopy (PAS). The demand for noninvasive blood glucose-level measurement has increased due to the explosive increase in diabetic patients. We have developed a noninvasive blood glucose-level measurement based on PAS. The conventional method uses a straight-type resonant cell. However, the cell volume is large, which results in a low detection sensitivity and difficult portability. In this paper, a small-sized Helmholtz-type resonant cell is proposed to improve detection sensitivity and portability by reducing the cell dead volume. First, the acoustic property of the small-sized Helmholtz-type resonant cell was evaluated by performing an experiment using a silicone rubber. As a result, the detection sensitivity of the small-sized Helmholtz-type resonant cell was approximately two times larger than that of the conventional straight-type resonant cell. In addition, the inside volume was approximately 30 times smaller. Second, the detection limits of glucose concentration were estimated by performing an experiment using glucose solutions. The experimental results showed that a glucose concentration of approximately 1% was detected by the small-sized Helmholtz-type resonant cell. Although these results on the sensitivity of blood glucose-level measurement are currently insufficient, they suggest that miniaturization of a resonance cell is effective in the application of noninvasive blood glucose-level measurement.

Mutation in insulin receptor attenuates oxidative stress and apoptosis in pancreatic beta-cells induced by nutrition excess: reduced insulin signaling and ROS.

Type 2 diabetes results from the failure of beta-cells to adequately compensate for insulin resistance. Although the reduction of beta-cell mass is because of increased cell death and/or inadequate replication or neogenesis, the mechanism underlying beta-cell mass reduction is not fully understood. Here, we clarify the role of insulin signaling pathway in the beta-cell apoptosis using insulin resistant model mice. Wild-type mice and those carrying a mutation in the insulin receptor (mIR) were fed either regular chow or a high-fat diet for 6 weeks and subsequently investigated for beta-cell apoptosis, endoplasmic reticulum stress, and oxidative stress. Insulin tolerance tests revealed that mIR mice fed a high-fat diet (mIRHF) had higher insulin resistance. Beta-cell apoptosis was increased 2-fold in the wild-type mice fed a high-fat diet (wHF) compared with control mice, whereas beta-cell apoptosis in mIRHF mice did not increase compared with that in mIR mice. The expression of endoplasmic reticulum stress markers in isolated islets did not differ between the groups. Staining of 8-hydroxy-2'-deoxyguanosine and 4-hydroxy-2-nonenal in islets of wHF mice significantly increased, but the staining in mIRHF mice was not different from that in control group. Gene expression of the antioxidant enzyme MnSOD was significantly higher in mIRHF mice than those in the other 3 groups. A mutation in the insulin receptor attenuated the oxidative stress and apoptosis in beta-cells even though high caloric nutrient was loaded. Our results suggest that reduced insulin signaling protects beta-cells thorough decline of oxidative stress.

Prevalence and risk factors of soil-transmitted helminth infection in Nepal.

BACKGROUND: The purpose of this study is to evaluate the prevalence and intensity and examine the risk factors of soil transmitted helminth (STH; i.e., roundworm [Ascaris lumbricoides], hookworms [Ancylostoma duodenale and Necator americanus], and whipworm [Trichuris trichiura]) infections in Nepal. METHODS: Five hundred and ninety-four adults (256 men and 338 women) were selected via convenience sampling from five communities in Nepal. The Kato-Katz method was used to assess the prevalence and intensity of STH infection in this population. RESULTS: Prevalence of STH infection ranged from 3.3% in Birendranagar in Chitwan, 3.5% in Kuleshor in Kathmandu, 11.7% in Kanyam in Ilam, 17.0% in Dhikurpokhari in Kaski and 51.4% in Khokana in Lalitpur District [corrected]. Multivariable regression analysis revealed that not using soap for hand-washing was significantly associated with the prevalence and infection intensity of roundworm, hookworms and whipworm. Similarly, not wearing sandals or shoes outside was significantly associated with the prevalence and infection intensity of roundworm and hookworms, but not with infection intensity of whipworm. Literacy, being underweight or overweight, anemia and occupation were not associated with prevalence and intensity of roundworm and hookworms infection, but there was an association between occupation and the prevalence of whipworm infection. CONCLUSION: STH infection was associated with individual hygiene behavior, but not with nutritional status or socio-demographic characteristics. Health policy focusing on changing individual hygiene behaviors might be useful in addressing STH infection in Nepal.

Polymyxin-B hemoperfusion for acute exacerbation of idiopathic pulmonary fibrosis: serum IL-7 as a prognostic marker.

BACKGROUND: Acute exacerbation (AE) of idiopathic pulmonary fibrosis (IPF) has an extremely poor prognosis. Direct hemoperfusion with a polymyxin B-immobilized fiber column (PMX-DHP) has been used to improve oxygenation for acute respiratory distress syndrome. The study aim was to retrospectively determine the predictive factors affecting the prognosis of AE of IPF treated with PMX-DHP. METHODS: We studied patients suffering from AE of IPF, treated with PMX-DHP combined with high-dose corticosteroid therapy. Stored serum taken before and after PMX-DHP therapy was analyzed for 27 cytokines and chemokines. RESULTS: Nineteen patients with AE of IPF were studied. The median survival time after diagnosis of AE was 22 days. Survival rates after diagnosis of AE were 47.4% at 30 days, 31.6% at 60 days, and 26.3% at 90 days. Serum levels of Interleukin (IL)-7, an anti-fibrotic cytokine, in survivors at day 30 following PMX-DHP therapy ('Survivors') significantly increased after the treatment, compared to serum levels of non-survivors at day 30 after the therapy ('Nonsurvivors'), which did not demonstrate a significant change. Serum levels of IL-1beta, interferon-y and chemokine ligand (CCL) 2 levels were not significantly altered in 'Survivors', but were significantly changed in 'Nonsurvivors.' Multivariate Cox proportional-hazards analysis showed that an increase in IL-7 levels after PMX-DHP therapy and treatment without intubation (other than invasive positive-pressure ventilation) were significantly better prognostic factors. CONCLUSION: The results suggest that serum IL-7 may be a useful prognostic factor for patients with AE of IPF treated with PMX-DHP, possibly reflecting underlying anti-fibrotic mechanisms.

Health status of married women residing five communities in Nepal: unexpectedly high prevalence of anemia in a well-off community of Kathmandu.

The high prevalence of anemia and underweight among reproductive age women is a serious health concern in Nepal. The objective of the present study was to describe anemia prevalence and nutritional status of married women in Nepal. Total of 278 women were included in the present analysis. The five communities included an urban commercial area (W) in Kathmandu, an agricultural village (K) in Lalitpur District, an agricultural village in Chitwan District (C) and one in Ilam District (I) with intensive cash cropping, and another rural village in Kaski District (P) with high number of international out-migration of males. The study procedure included anthropometric measurements, blood collection for hemoglobin measurement, and stool collection for examination of hookworm infection, and a structured interview on their reproductive history including contraceptive use. The altitude adjusted prevalence of anemia was 65%, 23%, 16%, 49% and 58% in communities C, I, K, P and W respectively. The hookworm prevalence of each community was not associated with the anemia prevalence. In the urban well-off community W, although 38% of the women had BMI > or = 25 and only 2% of women had BMI<18.5, 58% of the women were anemic. A logistic regression analysis on the anemia risk at the individual level showed no effect of helminth infection, but a significant negative effect of Depo-Provera (depot-medroxyprogesterone acetate) use. Causes of high anemia prevalence among the women resided in the urban well-off area should be investigated in future studies.

[Subscapular elastofibroma].

Elastofibroma is a tumor that is localized mainly at the subscapular region. We report 2 cases of subscapular elastofibromas. Case 1, 75-year-old woman was seen at the hospital because of a left dorsal tumor. Computed tomography (CT) scan revealed the tumor of 6 cm in diameter in the inferior angle of left scapula. The patient underwent excision of the tumor. Case 2, 90-year-old man underwent excision a tumor of 5 cm in diameter in the inferior angle of right scapula simultaneously with the operation of right lung cancer. Histological examinations showed increased elastic fiber with elastica van Gieson staining. These specimens confirmed the diagnosis of elastofibroma There have been no signs of recurrence after surgery.

Effects of open lung approach policy on mechanical ventilation duration in postoperative patients with chronic thromboembolism with pulmonary hypertension: a case-matched study.

Patients with chronic thromboembolism with pulmonary hypertension (CTEPH) often develop severe hypoxaemia after pulmonary thromboendarterectomy. There is reluctance to apply high positive end-expiratory pressure (PEEP) to those patients, whereas high PEEP is important for acute hypoxaemic respiratory failure due to alveolar collapse. Open lung approach (OLA) policy, a combination of recruitment manoeuvre and PEEP titration, may improve oxygenation and lung mechanics in acute hypoxaemic respiratory failure, but the effect of OLA on the outcome is unknown. We designed a case-matched, retrospective study to investigate whether OLA policy improved the outcome of postoperative patients with CTEPH. Among 113 postoperative patients with CTEPH, 40 were chosen before and after the introduction of an OLA policy to create 1:1 ratio of case-match according to gender, age and preoperative total pulmonary resistance index (conventional treatment group vs. OLA group). In the OLA group, recruitment manoeuvre was applied and then PEEP was titrated to maintain oxygenation every 12 hours. Gas exchange and duration of mechanical ventilation were compared between the groups. The OLA group showed higher PaO2/FiO2 ratio at 12 hours after the surgery than the conventional group (P = 0.0021). In the OLA group, duration of mechanical ventilation was shorter than the conventional treatment group (median, 23.5 hours vs. 43 hours, P = 0.0064). The OLA group showed lower cardiac index, higher pulmonary artery pressure and higher total pulmonary resistance index after the surgery than the conventional group. The introduction of the OLA policy may have shortened mechanical ventilation duration despite what appeared to be less favorable early postoperative hemodynamics in patients after the surgery for CTEPH.

MEK-ERK-dependent multiple caspase activation by mitochondrial proapoptotic Bcl-2 family proteins is essential for heavy ion irradiation-induced glioma cell death.

Recently developed heavy ion irradiation therapy using a carbon beam (CB) against systemic malignancy has numerous advantages. However, the clinical results of CB therapy against glioblastoma still have room for improvement. Therefore, we tried to clarify the molecular mechanism of CB-induced glioma cell death. T98G and U251 human glioblastoma cell lines were irradiated by CB, and caspase-dependent apoptosis was induced in both cell lines in a dose-dependent manner. Knockdown of Bax (BCL-2-associated X protein) and Bak (BCL-2-associated killer) and overexpression of Bcl-2 or Bcl-xl (B-cell lymphoma-extra large) showed the involvement of Bcl-2 family proteins upstream of caspase activation, including caspase-8, in CB-induced glioma cell death. We also detected the activation of extracellular signal-regulated kinase (ERK) and the knockdown of ERK regulator mitogen-activated protein kinase kinase (MEK)1/2 or overexpression of a dominant-negative (DN) ERK inhibited CB-induced glioma cell death upstream of the mitochondria. In addition, application of MEK-specific inhibitors for defined periods showed that the recovery of activation of ERK between 2 and 36 h after irradiation is essential for CB-induced glioma cell death. Furthermore, MEK inhibitors or overexpression of a DN ERK failed to significantly inhibit X-ray-induced T98G and U251 cell death. These results suggested that the MEK-ERK cascade has a crucial role in CB-induced glioma cell death, which is known to have a limited contribution to X-ray-induced glioma cell death.

Whether or not prophylactic excision of the extrahepatic bile duct is appropriate for patients with pancreaticobiliary maljunction without bile duct dilatation.

BACKGROUND/AIMS: The standard treatment for patients with a pancreaticobiliary maljunction (PBM) without bile duct dilatation remains controversial. METHODOLOGY: We followed up 29 patients with such PBM who mainly underwent a cholecystectomy alone. The ages of the patients ranged from 3 to 76 years (average age 47.3 years) and the ratio of males to females was 8 vs. 21. When the diameter of the common bile duct was less than 10mm, such bile ducts were diagnosed to have no dilatation. The main clinical indications for surgery were cholecystolithiasis in 15 patients, choledocholithiasis in 3, cholecystocholedocholithiasis in 2, gallbladder polyp in 2, adenomyomatosis in 2, cholecystitis in 2, and protein plug in 1. RESULTS: The amylase levels of gallbladder bile in 20 patients ranged from 115 to 460,200 IU/mL (a mean of 191,698 IU/mL). One patient died of gastric cancer 182 months after surgery and two patients died of other diseases 153, 171 months after surgeries, respectively. The remaining 26 patients have all been doing well for 36 months to 326 months after surgery (a median follow-up period, 160.5 months). The 10- and 15-year survival rates were 100% and 89.7%. CONCLUSIONS: In conclusion, a prophylactic resection of the extrahepatic bile duct and biliary diversion could be unnecessary for patients with PBM without bile duct dilatation.

Transdermal drug delivery using ultrasound-theory, understanding and critical analysis.

This review focuses on a unique transdermal drug delivery enhanced by the action of ultrasound, referred as sonophoresis. Sonophoresis is an active form of transdermal delivery which enhances the transport of permeants, such as drugs through cell membranes as a result of ultrasonic energy. Ultrasonic sound waves cause acoustic cavitation, the resultant effects of which microscopically disrupt the lipid bilayers of the stratum corneum and thereby influencing the influx of permeants. Sonophoresis increases the penetration of various low molecular weight drugs as well as high molecular weight proteins. The objective of this review is to account the role of ultrasound parameters and the associated cavitational effects, gained through a number of investigations, in order to facilitate the understanding of this method.

The RASSF1A isoform of RASSF1 promotes microtubule stability and suppresses tumorigenesis.

The RASSF1A isoform of RASSF1 is frequently inactivated by epigenetic alterations in human cancers, but it remains unclear if and how it acts as a tumor suppressor. RASSF1A overexpression reduces in vitro colony formation and the tumorigenicity of cancer cell lines in vivo. Conversely, RASSF1A knockdown causes multiple mitotic defects that may promote genomic instability. Here, we have used a genetic approach to address the function of RASSF1A as a tumor suppressor in vivo by targeted deletion of Rassf1A in the mouse. Rassf1A null mice were viable and fertile and displayed no pathological abnormalities. Rassf1A null embryonic fibroblasts displayed an increased sensitivity to microtubule depolymerizing agents. No overtly altered cell cycle parameters or aberrations in centrosome number were detected in Rassf1A null fibroblasts. Rassf1A null fibroblasts did not show increased sensitivity to microtubule poisons or DNA-damaging agents and showed no evidence of gross genomic instability, suggesting that cellular responses to genotoxins were unaffected. Rassf1A null mice showed an increased incidence of spontaneous tumorigenesis and decreased survival rate compared with wild-type mice. Irradiated Rassf1A null mice also showed increased tumor susceptibility, particularly to tumors associated with the gastrointestinal tract, compared with wild-type mice. Thus, our results demonstrate that Rassf1A acts as a tumor suppressor gene.

Longitudinal growth and height velocity of Japanese children with Down's syndrome.

AIM: To determine the natural growth pattern of Japanese children with Down's syndrome. METHODS: Longitudinal height data of 85 patients (43 males, 42 females) from birth to final height were analyzed. Based on these data, semi-longitudinal standard growth curves and height velocity curves for Down's syndrome were drawn. RESULTS: The means +/- SD of final height of males and females with Down's syndrome were 153.2 +/- 5.6 and 141.9 +/- 4.2 cm, respectively. They were -3.0 SD and -2.8 SD for Japanese standards. Mean peak height velocities were 8.9 and 7.5 cm y(-1), and the ages at peak height velocity were 11.6 and 10.2 y for males and females, respectively. CONCLUSION: The mean height of patients with Down's syndrome was around -2 SD for normal children before puberty. Their pubertal growth spurt starts about 1 y earlier and their peak height velocity was about 1.3-1.4 cm shorter than for normal children.

Proteases augment the effects of lipopolysaccharide in rat gingiva.

OBJECTIVES: Lipopolysaccharide (LPS) and proteases have been implicated as important factors in the initiation and progression of human periodontal diseases. A single application of LPS or proteases is insufficient to induce periodontal pocket formation or periodontitis. The aim of the present study was to assess the combined effect of lipopolysaccharide and proteases on rat periodontal tissues, and create a periodontal disease model. MATERIALS AND METHODS: Forty male Wistar rats were divided into four groups: combination group (treated with both LPS and proteases solutions); LPS group; proteases group; and control. Each solution was introduced daily into the palatal gingival sulcus of maxillary molars for 8 weeks. The tissues were evaluated histometrically and immunohistochemically. RESULTS: In the LPS group, elongation of rete ridge, apical migration of junctional epithelium (JE), increased numbers of B cells in connective tissue, and resorption of alveolar bone were observed. In the proteases group, the increase in the number of infiltrating polymorphonuclear leukocytes and blood vessels in the connective tissue was greater than that of the LPS group. CONCLUSIONS: The effects of LPS on periodontal tissues differed from those of proteases. The addition of proteases augmented and increased the effects of LPS, which were apical migration, intraepithelial cleavage of JE, and increased B cell density. The lesions in the combination group resembled established lesions of human periodontitis, with the exception of the low density of plasma cells.

Final height and pubertal growth in Japanese patients with congenital hypothyroidism detected by neonatal screening.

AIM: To investigate the final adult heights and pubertal growth patterns in Japanese patients with congenital hypothyroidism (CH) detected by neonatal screening. METHODS: A retrospective chart review was conducted of female patients >15 y of age (n = 18) and male patients >18 y of age (n = 9), who were detected by neonatal screening and kept on continuous thyroid hormone replacement therapy. Final height standard deviation scores (FHSDS) and target height standard deviation scores (THSDS) were determined. Parameters characterizing the pubertal growth process (such as age at onset of pubertal growth spurt and age at peak pubertal growth) were obtained from each patient's growth rate chart. Menarchial age was determined in each female patient by reviewing the medical record. The impact on FHSDS of the etiology of CH, the severity of CH, the time of initiation of therapy and the adequacy of treatment during the first year of life was assessed. RESULTS: All patients had received initial thyroid hormone treatment no later than 50 d of age, and had reached their final height. The mean FHSDS for female and male patients were +0.17 +/- 0.99 and -0.03 +/- 0.99, respectively. The mean FHSDS-THSDS for female and male patients was +0.09 +/- 0.77 and -0.19 +/- 0.53, respectively. No difference was seen in pubertal growth parameters for either gender compared with that of the reference population, except for a greater peak height velocity and pubertal height gain in male patients. The mean menarchial age was identical to that of the reference population. No significant relationship was found between the FHSDS and any of the factors investigated. CONCLUSION: The adult height of patients with CH detected by neonatal screening was equivalent to that of the reference population and their target height. As long as early intervention and satisfactory management are ensured, severe CH does not appear to reduce final adult height.

Effects of epinephrine and phosphodiesterase III inhibitors on bupivacaine-induced myocardial depression in guinea-pig papillary muscle.

BACKGROUND: This study was designed to investigate the effects of epinephrine and the phosphodiesterase III inhibitors amrinone and milrinone on bupivacaine-induced myocardial depression in guinea-pig papillary muscles using an electrophysiological method. METHODS: Electrophysiological studies of the effects of bupivacaine, epinephrine, amrinone and milrinone in normal and high K(+) Tyrode's solution were measured with guinea-pig papillary muscles. Specifically, epinephrine, amrinone and milrinone reversal of bupivacaine-induced depression was measured. RESULTS: Bupivacaine reduced the action potential duration (APD), the maximum rate of rise of the AP (V(max)) and contractile force. Although epinephrine increased the contractile force similarly to amrinone and milrinone, it shortened the APD at 50% repolarization (APD(50)) and 90% repolarization (APD(90)). A high concentration of amrinone shortened APD, while milrinone did not affect APD except for a prolongation of APD(20). In high K(+) Tyrode's solution (25 mM), epinephrine, amrinone and milrinone increased the APD and the contractile force. Epinephrine reversed bupivacaine depression of APD and contractile force to control levels. Amrinone and milrinone restored not only the contractile force but also APD. There was an incomplete recovery of APD(50) for amrinone and the prolongation of APD(20) for milrinone. CONCLUSIONS: Our results suggest that bupivacaine decreases the Ca(+) current (I(Ca)) and Na(+) current (I(Na)). Epinephrine and amrinone may increase I(Ca) and the delayed outward current (I(k)), whereas milrinone may increase I(Ca). The profound cardiovascular depression caused by bupivacaine was effectively reversed by amrinone and milrinone in a manner similar to epinephrine.

Laser excitation of high-frequency capillary waves.

The laser-induced surface deformation (LISD) technique was applied to generate high-frequency capillary waves on liquid surfaces up to several tens of kHz in a noncontact manner. The dynamic response of the fluid near the surface was theoretically derived under the condition of periodical radiation pressure. The result of the numerical calculation predicts the propagation of induced capillary waves out from the excitation region. The efficiency of the wave generation was experimentally examined by changing the width of the excitation laser beam at the surface. The observed LISD spectra were well reproduced by the theory, showing that the effective frequency band can be extended up to over 100 kHz. The propagation of the optically generated wave was measured with a laser probe sweeping the position of the observation. The spatial profile gives the surface tension and the shear viscosity of the sample liquid. The frequency domain measurement was also carried out and the spectrum obtained at a fixed point agrees with the theory, demonstrating the rapid measurement of frequency-dependent phenomena.

Development of safe and efficient novel nonviral gene transfer using ultrasound: enhancement of transfection efficiency of naked plasmid DNA in skeletal muscle.

Although clinical trials of stimulation of angiogenesis by transfection of angiogenic growth factors using naked plasmid DNA or adenoviral vector have been successful, there are still unresolved problems for human gene therapy such as low transfection efficiency and safety. From this viewpoint, it is necessary to develop safe and efficient novel nonviral gene transfer methods. As therapeutic ultrasound induces cell membrane permeabilization, ultrasound irradiation might increase the transfection efficiency of naked plasmid DNA into skeletal muscle. Thus, we examined the transfection efficiency of naked plasmid DNA using ultrasound irradiation with echo contrast microbubble (Optison) in vitro and in vivo experiments. First, we examined the feasibility of ultrasound-mediated transfection of naked plasmid DNA into skeletal muscle cells. Luciferase plasmid mixed with or without Optison was transfected into cultured human skeletal muscle cells using ultrasound (1 MHz; 0.4 W(2)) for 30 s. Interestingly, luciferase activity was markedly increased in cells treated with Optison, while little luciferase activity could be detected without Optison (P < 0.01). Electron microscopy demonstrated the transient formation of holes (less than 5 microM) in the cell surface, which could possibly explain the rapid migration of the transgene into the cells. Next, we studied the in vivo transfection efficiency of naked plasmid DNA using ultrasound with Optison into skeletal muscle. Two days after transfection, luciferase activity in skeletal muscle transfected with Optison using ultrasound was significantly increased about 10-fold as compared with plasmid alone. Successful transfection was also confirmed by beta-galactosidase staining. Finally, we examined the feasibility of therapeutic angiogenesis using naked hepatocyte growth factor (HGF) plasmid in a rabbit ischemia model using the ultrasound-Optison method. Five weeks after transfection, the angiographic score and the number of capillary density in rabbits transfected with Optison using ultrasound was significantly increased as compared with HGF plasmid alone (P < 0.01), accompanied by a significant increase in blood flow and blood pressure ratio (P < 0.01). Overall, the ultrasound transfection method with Optison enhanced the transfection efficiency of naked plasmid DNA in vivo as well as in vitro. Transfection of HGF plasmid by the ultrasound-Optison method could be useful for safe clinical gene therapy to treat peripheral arterial disease without a viral vector system.