Redox-labelled electrochemical aptasensors with nanosupported cancer cells.

The transfer of redox-labelled bioelectrochemical sensors from proteins to cells is not straightforward because of the cell downward force issue on the surface of the sensors. In this paper, 20-nm-thick nanopillars are introduced to overcome this issue, in a well-controlled manner. We show on both molecular dynamics simulations and experiments that suspending cells a few nanometers above an electrode surface enables redox-labelled tethered DNA aptamer probes to move freely, while remaining at an interaction distance from a target membrane protein, i. e. epithelial cell adhesion molecule (EpCAM), which is typically overexpressed in cancer cells. By this nanopillar configuration, the interaction of aptamer with cancer cells is clearly observable, with 13 cells as the lower limit of detection. Nanoconfinement induced by the gap between the electrode surface and the cell membrane appears to improve the limit of detection and to lower the melting temperature of DNA aptamer hairpins, offering an additional degree of freedom to optimize molecular recognition mechanisms. This novel nanosupported electrochemical DNA cell sensor scheme including Brownian-fluctuating redox species opens new opportunities for the design of all-electrical sensors using redox-labelled probes.

Selective layer-free blood serum ionogram based on ion-specific interactions with a nanotransistor.

Despite being ubiquitous in the fields of chemistry and biology, the ion-specific effects of electrolytes pose major challenges for researchers. A lack of understanding about ion-specific surface interactions has hampered the development and application of materials for (bio-)chemical sensor applications. Here, we show that scaling a silicon nanotransistor sensor down to ~25 nm provides a unique opportunity to understand and exploit ion-specific surface interactions, yielding a surface that is highly sensitive to cations and inert to pH. The unprecedented sensitivity of these devices to Na+ and divalent ions can be attributed to an overscreening effect via molecular dynamics. The surface potential of multi-ion solutions is well described by the sum of the electrochemical potentials of each cation, enabling selective measurements of a target ion concentration without requiring a selective organic layer. We use these features to construct a blood serum ionogram for Na+, K+, Ca2+ and Mg2+, in an important step towards the development of a versatile, durable and mobile chemical or blood diagnostic tool.

In situ regeneration of retinal pigment epithelium by gene transfer of E2F2: a potential strategy for treatment of macular degenerations.

The retinal pigment epithelium (RPE) interacts closely with photoreceptors to maintain visual function. In degenerative diseases such as Stargardt disease and age-related macular degeneration, the leading cause of blindness in the developed world, RPE cell loss is followed by photoreceptor cell death. RPE cells can proliferate under certain conditions, suggesting an intrinsic regenerative potential, but so far this has not been utilised therapeutically. Here, we used E2F2 to induce RPE cell replication and thereby regeneration. In both young and old (2 and 18 month) wildtype mice, subretinal injection of non-integrating lentiviral vector expressing E2F2 resulted in 47% of examined RPE cells becoming BrdU positive. E2F2 induced an increase in RPE cell density of 17% compared with control vector-treated and 14% compared with untreated eyes. We also tested this approach in an inducible transgenic mouse model of RPE loss, generated through activation of diphtheria toxin-A gene. E2F2 expression resulted in a 10-fold increase in BrdU uptake and a 34% increase in central RPE cell density. Although in mice this localised rescue is insufficiently large to be demonstrable by electroretinography, a measure of massed retinal function, these results provide proof-of-concept for a strategy to induce in situ regeneration of RPE for the treatment of RPE degeneration.

Effects of bisphosphonates on human esophageal squamous cell carcinoma cell survival.

Esophageal squamous cell carcinoma (ESCC) is one of the most malignant cancers in Japan. Anticancer chemotherapy has been useful for ESCC treatment. However, therapeutic options are limited. Recently, bisphosphonates (BPs), which are osteoporosis drugs, have shown anticancer effects in several cancer cell lines, but the effects against ESCC cell lines are unknown. In this study, we examined the cytotoxic effects of BPs and their mechanisms of cytotoxicity in human ESCC cell lines. A first-generation BP (etidronate), two second-generation BPs (alendronate and pamidronate), and two third-generation BPs (risedronate and zoledronate) were used in this study. All BPs, except etidronate, were cytotoxic, as indicated by increased caspase-3/7 activity and numbers of Annexin-fluorescein isothiocyanate positive cells in ESCC cell lines. From cell cycle analysis, G0/G1-phase arrest was observed upon treatment with second- and third-generation BPs. In addition, Cyclin D1 protein expression levels were decreased by second- and third-generation BP treatment. Although squalene and trans, trans-farnesol minimally affected BP cytotoxicity, treatment with geranylgeraniol inhibited BP cytotoxicity almost completely. We concluded that second- and third-generation BPs are cytotoxic to ESCC cell lines as they induce apoptosis and inhibit the cell cycle through mevalonate pathway inhibition. Therefore, BP treatment may be a beneficial therapy in ESCC patients.

Dispersive and dissipative coupling in a micromechanical resonator embedded with a nanomechanical resonator.

A micromechanical resonator embedded with a nanomechanical resonator is developed whose dynamics can be captured by the coupled-Van der Pol-Duffing equations. Activating the nanomechanical resonator can dispersively shift the micromechanical resonance by more than 100 times its bandwidth and concurrently increase its energy dissipation rate to the point where it can even be deactivated. The coupled-Van der Pol-Duffing equations also suggest the possibility of self-oscillations. In the limit of strong excitation for the nanomechanical resonator, the dissipation in the micromechanical resonator can not only be reduced, resulting in a quality factor of >3× 10(6), it can even be eliminated entirely resulting in the micromechanical resonator spontaneously vibrating.

Electric tuning of direct-indirect optical transitions in silicon.

Electronic band structures in semiconductors are uniquely determined by the constituent elements of the lattice. For example, bulk silicon has an indirect bandgap and it prohibits efficient light emission. Here we report the electrical tuning of the direct/indirect band optical transition in an ultrathin silicon-on-insulator (SOI) gated metal-oxide-semiconductor (MOS) light-emitting diode. A special Si/SiO2 interface formed by high-temperature annealing that shows stronger valley coupling enables us to observe phononless direct optical transition. Furthermore, by controlling the gate field, its strength can be electrically tuned to 16 times that of the indirect transition, which is nearly 800 times larger than the weak direct transition in bulk silicon. These results will therefore assist the development of both complementary MOS (CMOS)-compatible silicon photonics and the emerging "valleytronics" based on the control of the valley degree of freedom.

A multimode electromechanical parametric resonator array.

Electromechanical resonators have emerged as a versatile platform in which detectors with unprecedented sensitivities and quantum mechanics in a macroscopic context can be developed. These schemes invariably utilise a single resonator but increasingly the concept of an array of electromechanical resonators is promising a wealth of new possibilities. In spite of this, experimental realisations of such arrays have remained scarce due to the formidable challenges involved in their fabrication. In a variation to this approach, we identify 75 harmonic vibration modes in a single electromechanical resonator of which 7 can also be parametrically excited. The parametrically resonating modes exhibit vibrations with only 2 oscillation phases which are used to build a binary information array. We exploit this array to execute a mechanical byte memory, a shift-register and a controlled-NOT gate thus vividly illustrating the availability and functionality of an electromechanical resonator array by simply utilising higher order vibration modes.

Water electrolysis and energy harvesting with zero-dimensional ion-sensitive field-effect transistors.

The relationship of the gas bubble size to the size distribution critically influences the effectiveness of electrochemical processes. Several optical and acoustical techniques have been used to characterize the size and emission frequency of bubbles. Here, we used zero-dimensional (0D) ion-sensitive field-effect transistors (ISFETs) buried under a microbath to detect the emission of individual bubbles electrically and to generate statistics on the bubble emission time. The bubble size was evaluated via a simple model of the electrolytic current. We suggest that energy lost during water electrolysis could be used to generate electric pulses at an optimal efficiency with an array of 0D ISFETs.

Bilateral central retinal artery occlusion and vein occlusion complicated by severe choroidopathy in systemic lupus erythematosus.

Severe retinal vascular occlusions resulting in blindness is a rare occurrence in patients with systemic lupus erythematosus (SLE). Herein, we report a case of a 33-year-old female who developed combined central retinal artery occlusion, retinal vein occlusion, and choroidopathy and rapidly became completely blind in both eyes within a week. The electroretinogram revealed a severely attenuated a-wave and b-wave, indicating a profound dysfunction of both choroidal and retinal circulation, respectively. The current case demonstrates objectively the functional impact of severe choroidopathy in SLE for the first time. Patients with unilateral blindness due to combined retinal/choroidal vascular obstructions should be monitored carefully to ensure adequate anticoagulant therapy in an attempt to guard the vision in the fellow eye.

Phonon lasing in an electromechanical resonator.

An electromechanical resonator harboring an atomlike spectrum of discrete mechanical vibrations, namely, phonon modes, has been developed. A purely mechanical three-mode system becomes available in the electromechanical atom in which the energy difference of the two higher modes is resonant with a long-lived lower mode. Our measurements reveal that even an incoherent input into the higher mode results in coherent emission in the lower mode that exhibits all the hallmarks of phonon lasing in a process that is reminiscent of Brillouin lasing.

Interconnect-free parallel logic circuits in a single mechanical resonator.

In conventional computers, wiring between transistors is required to enable the execution of Boolean logic functions. This has resulted in processors in which billions of transistors are physically interconnected, which limits integration densities, gives rise to huge power consumption and restricts processing speeds. A method to eliminate wiring amongst transistors by condensing Boolean logic into a single active element is thus highly desirable. Here, we demonstrate a novel logic architecture using only a single electromechanical parametric resonator into which multiple channels of binary information are encoded as mechanical oscillations at different frequencies. The parametric resonator can mix these channels, resulting in new mechanical oscillation states that enable the construction of AND, OR and XOR logic gates as well as multibit logic circuits. Moreover, the mechanical logic gates and circuits can be executed simultaneously, giving rise to the prospect of a parallel logic processor in just a single mechanical resonator.

One-by-one trap activation in silicon nanowire transistors.

Flicker or 1/f noise in metal-oxide-semiconductor field-effect transistors (MOSFETs) has been identified as the main source of noise at low frequency. It often originates from an ensemble of a huge number of charges becoming trapped and de-trapped. However, as a deviation from the well-known model of 1/f noise is observed for nanoscale MOSFETs, a new model is required. Here, we report the observation of one-by-one trap activation controlled by the gate voltage in a nanowire MOSFET and propose a new low-frequency-noise theory for nanoscale FETs. We show that the Coulomb repulsion between electronically charged trap sites prevents the activation of several traps simultaneously. This effect induces a noise reduction of more than one order of magnitude. It decreases when the electron density in the channel is increased due to the electrical screening of traps. These findings are technologically useful for any FET with a short and narrow channel.

Single-electron counting statistics and its circuit application in nanoscale field-effect transistors at room temperature.

A circuit utilizing single electrons is demonstrated at room temperature. Individual electrons randomly passing through the nanoscale silicon-on-insulator metal-oxide-semiconductor field-effect transistor (MOSFET) are monitored by an electrometer in real time. Such a random behavior of single electrons is used for high-quality random-number generation suitable for data processing which stochastically extracts the most preferable pattern among various ones. MOSFET-based random-number generation allows fast operation as well as high controllability, which leads to flexible extraction of the preferable pattern.

Effects of ABCB1 3435C>T genotype on serum levels of cortisol and aldosterone in women with normal menstrual cycles.

ABCB1, also known as MDR1/P-glycoprotein, can transport cortisol and aldosterone. We examined the effects of ABCB1 polymorphisms on serum levels of cortisol and aldosterone among different phases of the normal menstrual cycle in 51 non-pregnant healthy Japanese female volunteers (22 +/- 1 years old). The menstrual cycle was divided into three phases: premenstrual phase (14 days preceding the onset of menstruation, N = 22; menstrual phase, N = 11, and postmenstrual phase, N = 18). ABCB1 -129T>C, 1236C>T, 2677G>A/T, and 3435C>T genotypes were determined. Serum levels of cortisol, aldosterone, estradiol, progesterone, and testosterone were measured. The serum levels of estradiol in the pre- and post-menstrual phases and of progesterone in the premenstrual phase were significantly increased when compared to their serum levels in the menstrual phase (P < 0.005). In the postmenstrual phase, the mean serum cortisol level in subjects with the 3435CT and 3435TT genotype was 7.6 +/- 3.4 microg/dL (mean +/- SD, N = 7), which was significantly lower than in women with the 3435CC genotype (9.9 +/- 1.8 microg/dL, N = 11) (P = 0.037). The opposite effect was observed in the serum aldosterone level during the postmenstrual phase (97.2 +/- 23.4 and 141.2 +/- 48.5 pg/mL for 3435CC and 3435CT + 3435TT, respectively; P = 0.041). These findings suggest that ABCB1 3435C>T genotype can influence serum levels of cortisol and aldosterone during the postmenstrual phase of the normal menstrual cycle.

Recessive mutations in the CYP4V2 gene in East Asian and Middle Eastern patients with Bietti crystalline corneoretinal dystrophy.

BACKGROUND: Bietti crystalline corneoretinal dystrophy (BCD) is an autosomal recessively inherited disorder characterised by tiny yellowish glittering retinal crystals, choroidal sclerosis, and crystals in the peripheral cornea, associated with progressive night blindness. CYP4V2, encoding a member of cytochrome p450 (CYP450) protein family, was recently identified as the causative gene. METHODS: We recruited 11 unrelated patients with BCD and characteristic clinical features; eight of Japanese, two of Middle Eastern, and one of Chinese ancestry. Genomic DNA was extracted from peripheral blood leucocytes, and all 11 exons and the flanking intron splice sites of the CYP4V2 gene were amplified and sequenced. A complete ophthalmological examination was performed. RESULTS: We found recessive mutations in the CYP4V2 gene in all of the 11 patients. Two novel mutations, L173W and Q450X, were identified in a Japanese patient and two unrelated patients from Middle Eastern countries, respectively. Each patient was a homozygote. A previously reported mutation IVS6-8_810delinsGC was identified in seven unrelated Japanese patients and the Chinese patient with BCD. All patients with BCD shared a characteristic fundus appearance with numerous intraretinal crystal deposits and atrophy of the retinal pigment epithelium. However, the clinical findings, including elecroretinograph recordings, indicated that there was considerable variation in the degree of visual dysfunction even among patients of similar ages carrying the same mutation. CONCLUSIONS: Defects in CYP4V2 are the main cause of BCD. The IVS6-8_810delinsGC mutant allele may be especially prevalent among patients with BCD in East Asian countries, resulting from a single founder. Variation of disease severity suggests that environmental or additional genetic factors influence the course of the retinal disease.

Laparoscopic approach for early gastric cancer within a paraesophageal hiatal hernia.

We present a case of early gastric cancer located in gastric volvulus associated with paraesophageal hiatal hernia. Two lesions of EGC were diagnosed in the distal third of the stomach, most of which had herniated into the left chest through a large hiatal defect in an organoaxial fashion. Routinely, laparoscopic-assisted distal gastrectomy (LADG) is our preferred approach for EGC, and the presence of hiatal hernia in this case did not alter our approach. Laparoscopic repair of hiatal hernia was performed successfully followed by LADG. A review of the literature supports a minimally invasive approach for both procedures and shows it to be safe, effective, and technically feasible. Further, LADG is shown to be oncologically adequate in terms of tumor margins and lymph node dissection, but its relevance to long-term disease-free survival still needs to be studied in well-designed prospective trials.

Efficacy of intraperitoneal and intravenous chemotherapy and left upper abdominal evisceration for advanced gastric cancer.

BACKGROUND: The study was carried out to evaluate the efficacy of intraperitoneal (i.p.) and intravenous (i.v.) chemotherapy, as well as left upper abdominal evisceration (LUAE), for patients with advanced gastric cancer. METHODS: We carried out a retrospective study of 348 patients who underwent gastrectomy for advanced gastric carcinoma between 1978 and 1998 at our institution and who had macroscopic type 3 or 4 cancer (Japanese classification) with depth of invasion to the serosal surface, but no liver metastasis or lymph node metastasis around the abdominal aorta. Cumulative survival rates were compared in patients who underwent gastrectomy together with: (1) intraoperative i.p. chemotherapy alone, (2) postoperative i.v. chemotherapy alone, (3) both i.p. and i.v., or (4) no chemotherapy. Then patients were stratified according to the presence of peritoneal dissemination (P+) and its absence (P-). In P+ patients, survival was compared between those who received i.v. chemotherapy and those who did not, and between those who received i.p. chemotherapy and those who did not. Then, survival was compared between patients with high and low immunosuppressive acidic protein (IAP) levels. Finally, we compared cumulative survival rates in patients (stratified as P+ and P-) who underwent LUAE with cumulative survival rates in those who underwent total gastrectomy combined with resection of the pancreatic body, tail, and spleen (PS). RESULTS: For P- patients, there was no survival advantage with adjuvant i.p. or i.v. therapy when compared with surgery alone. For P+ patients, however, there was an improvement in survival when patients received both i.p. and i.v., compared with survival with surgery alone (P < 0.05). In P+ patients aged less than 60 years, there was improvement in survival for those who underwent i.p. therapy together with surgery (P < 0.05), but not for those who had i.v. chemotherapy after surgery. When LUAE was examined, there was a survival advantage for this procedure when there was no peritoneal dissemination. Four long-term survivors (surviving for more than 5 years) were identified in our study. Three of the 4 patients were aged less than 60 years, and all 4 had macroscopic type 4 gastric cancers. CONCLUSION: Although the prognosis for patients with invasive type gastric cancer remains poor, there have been a few long-term survivors, in whom this survival was associated with aggressive combination therapy, including surgery, i.p., and i.v. therapy. P+ patients aged less than 60 years and patients with type 4 gastric cancer may stand to benefit most from such therapy. For P- patients, the role of adjuvant i.p. or i.v. therapy continues to be ambiguous, although LUAE in this population may be superior to PS.

CYP2C19 genotype and pharmacokinetics of three proton pump inhibitors in healthy subjects.

PURPOSE: To predict the CYP2C19 genotype-dependence in anti-Helicobacter pylori (H. pylori) therapy when lansoprazole or rabeprazole was used instead of omeprazole as a proton pump inhibitor (PPI). METHODS: A comparative pharmacokinetic study with each PPI was designed as an open, randomized, and crossover study of 18 Japanese healthy volunteers who were classified into the homozygous, heterozygous extensive metabolizer and the poor metabolizer based on the CYP2C19 genotype determined by PCR-RFLP method. Each subject received a single oral dose of 20 mg omeprazole, 30 mg lansoprazole, or 20 mg sodium rabeprazole, with at least 1 week washout period between treatments. Plasma concentrations of PPIs and their metabolites were monitored until 12 h after medication. RESULTS: Pharmacokinetic profiles of omeprazole and lansoprazole were well correlated with the CYP2C19 genotype. The heterozygous extensive metabolizer was slightly different from the homozygote, but there was no statistically significant difference. The CYP2C19 genotype dependence found for lansoprazole was not obvious compared with omeprazole. As for rabeprazole, the pharmacokinetic profile was independent of the CYP2C19 genotype. CONCLUSIONS: CYP2C19 genotype dependence will be found in the anti-H. pylori therapy even when lansoprazole is used as the PPI.

Effect of serum triglyceride concentration on the fluctuation of whole blood concentration of cyclosporin A in patients.

The methodology to distinguish the patients showing considerable fluctuation of the whole blood concentration of cyclosporin A (CYA) was investigated from a viewpoint of laboratory test values. First, we retrospectively examined the CYA trough blood concentrations monitored continuously. The patients were classified into three groups by the fluctuation of CYA trough blood concentrations during the examination period (Cmax/Cmin): Group 1 (Cmax/Cmin=100-200%; n=21), Group 2 (Cmax/Cmin=200-300%; n=25), and Group 3 (Cmax/Cmin=more than 300%; n=32). In the laboratory tests examined, the serum triglyceride concentrations were considerably different among the groups, and it was the highest in Group 3. Next, to elucidate the effect of serum triglyceride concentration on the CYA blood concentration, in vivo pharmacokinetic studies after single intravenous or repetitive oral administration of CYA were conducted in the model rats with pseudo-hypertriglyceridemia, hypocythemia, and acute renal failure. Only in pseudo-hypertriglyceridemia rats, the CYA blood concentration after a single intravenous injection was significantly higher than that in normal rats because of the restriction of CYA distribution to the extravascular tissues. On the other hand, the increase in the serum triglyceride concentration did not affect the fluctuation of CYA trough blood concentration after repetitive oral administration. Taken together, the fluctuation of CYA trough blood concentrations observed in the clinical situation could be due to the fluctuation of serum triglyceride concentration, and the patients with such fluctuation of serum triglyceride concentrations might also be distinguishable by the higher concentration of serum triglyceride in laboratory tests.

Comparative evaluation of surgical stress of laparoscopic and open surgeries for colorectal carcinoma.

PURPOSE: To objectively evaluate the benefits of laparoscopic procedures for colorectal carcinoma, a prospective study to measure the stress response to laparoscopic surgery (n = 15) compared with open surgery (n = 12) was undertaken. In addition, to compare the various parameters relevant to surgical stress, the major surgery group (transthoracic esophagectomy for esophageal carcinoma, n = 4; and left upper abdominal evisceration for gastric carcinoma, n = 3) was assigned. METHODS: Peripheral blood samples were obtained to measure serum interleukin-6, C-reactive protein, peripheral leukocytes, and lymphocyte counts. Additionally, the level of lymphocyte apoptosis was quantified using flow cytometry. RESULTS: The interleukin-6 and C-reactive protein levels were significantly greater in the open group than in the laparoscopy group one day (P < 0.05) and two days (P < 0.05) after surgery, respectively. In the laparoscopy group, lymphocyte counts were significantly higher than in the open group two days after surgery. The laparoscopy and open groups did not differ significantly in their lymphocyte apoptotic index. In the major surgery group, the apoptotic index was significantly higher than in either the laparoscopy group or the open group in the early postoperative period. CONCLUSION: Changes in the various parameters pertinent to surgical stress evaluated in this study suggest that laparoscopic surgery for colorectal carcinoma leads to less postoperative stress than conventional open surgery.