The aim of this study was to assess whether a particular value of noninvasive salivary ultra-weak chemiluminescence (UCL) could be used as a biomarker of psychological stress. Our study covered two groups. Group 1 comprised six healthy volunteers who stayed in a hospital for one night and group 2 comprised 15 patients with lung cancer and 24 patients with respiratory diseases other than lung cancer who were in hospital for an extended stay. First, we evaluated the UCL of saliva from six healthy volunteers before and after one night in hospital. Immunoglobulin A (IgA) concentrations were also measured. The integrated intensity value of UCL was correlated with the IgA concentration (correlation coefficient 0.90). Second, in the case of a long hospital stay, we found that the maximum salivary UCL intensities were higher in patients with lung cancer than in those with respiratory diseases other than lung cancer or in 28 healthy controls. Copyright © 2016 John Wiley & Sons, Ltd.

Nanonization, which involves the formation of the drug with nanometer particle size, is an effective method to improve the dissolution rate and bioavailability of poorly water-soluble drugs. A pulsewidth-tunable femtosecond laser was used to produce nanoparticles of clobetasone butyrate using poloxamer 188as stabilizing agent. The effects of temperature and pulsewidth on the particle size and concentration were studied for the first time. The particle size and drug concentration dependence on the laser intensity and irradiation time were also investigated. Permeability test releaved that laser nanonization improved the drug permeability across Caco-2 cell monolayer. This laser nanonization method has a great potential to be used for new drug development.

Background: For monitoring biomarkers, saliva has several distinct advantages over other biological fluids. Saliva sampling is relatively non-invasive and enables the collection from either adults or infants under many different circumstances. However, there is no collection device that can be speedily used for analysis in the field. The aim of the present study was to evaluate the suitability of a new device, termed Muddler, compared with commercially available collection devices.Methods: Saliva was collected from healthy volunteers. The collection devices such as Muddler, eye sponge, Salivette (R) Cotton (SC) and Salivette (R) Synthetic (SS) were evaluated in terms of the volume and/or composition of the collected saliva. The amounts of immunoglobulin A (IgA) and lactofferin in saliva were assessed by the enzyme-linked immunosorbent assays with the corresponding antibodies. Amylase activity was measured using a commercially available kit, and high molecular weight complexes including mucin were assessed by SDS-PAGE staining.Results: A newly developed Muddler, which was made of plastic plate, was the best device for collecting a constant volume of saliva among all the devices examined in the present study. Furthermore, Muddler can collect without change in composition of salivary proteins such as IgA, lactoferrin, amylase, and mucin complex, whereas the levels of the salivary proteins obtained with all the commercial devices used were clearly different from those in original saliva.Conclusions: The newly developed Muddler was the best collection device in terms of the accuracy of collection and the reliability of measurements among all the devices examined in the present study. (C) 2013 Elsevier B.V. All rights reserved.

Terahertz pulsed spectroscopy has recently been demonstrated to be a novel technique for the investigation of the solid-state properties of pharmaceutical materials. In this study, we directly measured the crystallinity of a drug suspended in water, using a terahertz pulsed attenuated total reflection (ATR) method. The dihydropyridine calcium channel blocker nifedipine is classified as a poorly soluble drug; its most stable crystalline form is known as form I. Transmission spectra, collected from 0.2 to 2.0 THz (6.6 to 66 cm(-1) ), of nifedipine crystals had a strong absorption peak at 1.2 THz (40 cm(-1) ) at room temperature. When the nifedipine crystals were mixed with poloxamer 188 and suspended in water, the resulting spectra measured using the ATR method had a peak at the same frequency as in the spectra obtained in transmission mode. Furthermore, the peak area was proportional to the amount of crystals. The upward sloping baseline in the spectra, corresponding to water absorption, decreased stepwise with increasing amounts of crystalline particles. We confirmed that the spectra gave excellent quantitative results, using partial least-squares regression analysis. The results suggest the possibility of using this method for qualitative and quantitative assessments of crystalline drugs in suspension.

We have developed a measuring system for simultaneous monitoring of chemiluminescence and fluorescence, which indicate respectively, (i) generation of superoxide anion radicals (O2(-•)) and (ii) change in the intracellular calcium ion concentration ([Ca(2+)]i) of neutrophils triggered by the mechanism of innate immune response. We applied this measuring system for establishing a method to distinguish between anti-inflammatory actions and antioxidant actions caused by bioactive compounds. We evaluated anti-inflammatory agents (zinc ion [Zn(2+)] and ibuprofen) and antioxidants (superoxide dismutase [SOD] and ascorbic acid). It was shown that ibuprofen and Zn(2+) were anti-inflammatory while SOD and ascorbic acid were anti-oxidative. We conclude that it is possible to determine the mechanism of action of bioactive compounds using this method.

Nanoparticle formation of poorly water-soluble drugs is a means of providing much benefit for improving solubility and bioavailability. We showed that laser irradiation of drugs can be a novel tool for dispersing drug nanoparticles in water. Using our method, we were able to produce nanoparticles containing immunosuppressant drug, cyclosporin A, which shows poor solubility toward water, with high levels of the drug using polyvinyl pyrrolidone and sodium dodecyl sulfate as stabilizing agents. The absence of degradation products was confirmed and the loss of pharmaceutical activity with an inhibitory effect on the interleukin-2 production of Jurkat T cells did not occur. Cyclosporin A nanoparticles showed a spherical shape and their particle size was distributed uniformly around 200 nm. Powder X-ray diffraction analysis suggested that cyclosporin A in the nanoparticles was in an amorphous state. In the measurement of solubility rate, the nanoparticle formulation showed a higher rate than that which had not been processed. At present, although this laser irradiation technology has low productivity, it is expected as a new technology for drug nanoparticle manufacturing together with the development of a new laser device.

Oligohydramnios is often caused by the premature rupturing of membranes and subsequent intrauterine infections, such as chorioamnionitis, in which event oxidative stress is hypothesized to be closely associated with the damage to the fetal organs. The clinical efficiency of amnioinfusion using warmed saline in cases of premature rupture of membranes is still controversial, especially concerning the prognosis for the fetus. In the present study, we found that human amniotic fluid per se suppresses the release of superoxide from cultured human neutrophils, suggesting an acute or chronic shortage of amniotic fluid in cases of premature rupture of membranes can affect the shielding of intrauterine organs from oxidative stress. The aim of this study was to propose a formula of zinc and magnesium ions in saline for amnioinfusion, by assessing antioxidative activities. A combination of 5μM zinc and 5mM magnesium in saline synergistically inhibited superoxide production by cultured human neutrophils, equivalent to human amniotic fluid. The intraperitoneal administration of this formula significantly improved the survival rate in a rat model of peritonitis compared to the saline control (46.7% vs. 10%). The combination of these metals with saline may thus be a promising formula for an amnioinfusion fluid with the capacity to protect fetal organs from oxidative stress.

Melamine is a raw ingredient for the manufacture of plastics, but it is frequently misused by adding it to food to raise the nitrogen content, thereby giving the false impression of a high protein content. Varied amounts of melamine were added to samples of dried milk obtained from five manufacturers in Japan. The samples were illuminated by a small cross section of a laser beam and the scattered light was examined. The presence of melamine in milk could be immediately detected without any chemical or physical alteration of the milk by interpreting its Raman scattering spectra. Among the many Raman bands, an intense band at 676 cm−1 was the most useful for detecting melamine; the detection limit was about 1% (w/w). Because this method does not extract melamine from the dried milk sample, it reduces the risk of error that may occur during extraction or from interaction with chemical reagents. The method provides a very rapid screening test for melamineadulterated dried milk in food chemistry and forensic toxicology.

BACKGROUND:Gastric cancer is one of the most common cancers in Japan. The use of endoscopy is increasing, along with the number of histological examinations of specimens obtained by endoscopy. However, it takes several days to reach a diagnosis, which increases the medical expense. Raman spectroscopy is one of the available optical techniques, and the Raman spectrum for each molecule and tissue is characteristic and specific. The present study investigated whether Raman spectroscopy can be used to diagnose gastric cancer.METHODS:A total of 251 fresh biopsy specimens of gastric carcinoma and non-neoplastic mucosa were obtained from 49 gastric cancer patients at endoscopy. Without any pretreatment, the fresh specimens were measured with a near-infrared multichannel Raman spectroscopic system with an excitation wavelength of 1064 nm, and Raman spectra specific for the specimens were obtained. A principal component analysis (PCA) was performed to distinguish gastric cancer and non-neoplastic tissue, and a discriminant analysis was used to evaluate the accuracy of the gastric cancer diagnosis.RESULTS:The Raman spectra for cancer specimens differed from those for non-neoplastic specimens, especially at around 1644 cm(-1). Sensitivity was 66%, specificity was 73%, and accuracy was 70%. The accuracy of diagnosis using the single Raman scattering intensity at 1644 cm(-1) was 70%, consistent with the PCA result.CONCLUSIONS:The present results indicate that near-infrared multichannel Raman spectroscopy with a 1064-nm excitation wavelength is useful for gastric cancer diagnosis. Establishment of a Raman diagnostic system for gastric cancer may improve the clinical diagnosis of gastric cancer and be beneficial for patients.

Saliva sampling has the advantage of being noninvasive and stress free. Based on a recent study, salivary ultraweak chemiluminescence (UCL) is a new biomarker of psychologic stress. However, it is not clear what causes changes in the UCL level and whether the change is biologically significant. We investigated the candidates for salivary UCL induced by psychological stressors and discuss the physiologic function of these candidates. Volunteers completed a questionnaire and then performed the Kraepelin test. Saliva was sampled just before, immediately after, and 30 min after the stress exposure. The UCL of saliva significantly increased just after stress exposure (1.56-fold) and returned to prestress levels after 30 min. The concentration of secretory immunoglobulin A also increased significantly and the change in both biomarkers was rapid. Similar significant changes were observed in salivary peroxidase activity and the concentration of thiocyanate (SCN-). On the other hand, the levels of amylase activity did not significantly increase and the concentration of cortisol increased slowly. Moreover, in the reconstitution experiment, UCL was generated at the same level by a mixture of peroxidase and SCN- at physiologic concentrations. In conclusion, we determined that the Kraepelin test as a mental arithmetic task elicited a significant response in the body and this response can be calculated using salivary UCL. Furthermore, SCN- and peroxidase in the saliva play a key role in salivary UCL.