[Attempt to Calculate the Efficiency of Methylation Using 1H-NMR for Analyzing Brominated Vegetable Oil].

We studied the efficiency of methylation for analyzing brominated vegetable oil (BVO). In this report, we investigated whether 1H-NMR is an applicable method for assessing the efficiency of methylation to analyze BVO. 1H-NMR sufficiently calculated the efficiency of methylation using each integral and the numbers of protons derived from the methyl group, which is characteristic in products, and the methine group, which is characteristic in unreacted substances. Additionally, the efficiency of methylation calculated via 1H-NMR was in good agreement with changes in the peak area of BVO fatty acid methyl esters (BVOFAMEs) after various heating times obtained from GC-FID analysis. Therefore, 1H-NMR is applicable for calculating the efficiency of methylation to analyze BVO.

Arginine vasopressin altered the expression of monocarboxylate transporters in cultured astrocytes isolated from stroke-prone spontaneously hypertensive rats and congenic SHRpch1_18 rats.

BACKGROUND: Astrocytes support a range of brain functions as well as neuronal survival, but their detailed relationship with stroke-related edema is not well understood. We previously demonstrated that the release of lactate from astrocytes isolated from stroke-prone spontaneously hypertensive rats (SHRSP/Izm) was attenuated under stroke conditions. The supply of lactate to neurons is regulated by astrocytic monocarboxylate transporters (MCTs). The purpose of this study was to examine the contributions of arginine vasopressin (AVP) and/or hypoxia and reoxygenation (H/R) to the regulation of MCTs and neurotrophic factor in astrocytes obtained from SHRSP/Izm and congenic SHRpch1_18 rats. METHODS: We compared AVP-induced lactate levels, MCTs, and brain-derived neurotrophic factor (BDNF) in astrocytes isolated from SHRSP/Izm, SHRpch1_18, and Wistar Kyoto rats (WKY/Izm). The expression levels of genes and proteins were determined by PCR and Western blotting (WB). RESULTS: The production of lactate induced by AVP was increased in astrocytes from all three strains. However, the levels of lactate were lower in SHRSP/Izm and SHRpch1_18 animals compared with the WKY/Izm strain. Gene expression levels of Slc16a1, Slc16a4, and Bdnf were lowered by AVP in SHRSP/Izm and SHRpch1_18 rats compared with WKY/Izm. The increase of MCT4 that was induced by AVP was blocked by the addition of a specific nitric oxide (NO) chelator, 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (CPTIO). Furthermore, AVP increased the expression of iNOS and eNOS proteins in WKY/Izm and SHRSP/Izm rat astrocytes. However, the iNOS expression levels in SHRSP astrocytes differed from those of WKY/Izm astrocytes. The increase of MCT4 protein expression during AVP treatment was blocked by the addition of a specific NF-kB inhibitor, pyrrolidine dithiocarbamate (PDTC). The induction of MCT4 by AVP may be regulated by NO through NF-kB. CONCLUSIONS: These results suggest that the expression of MCTs mediated by AVP may be regulated by NO. The data suggest that AVP attenuated the expression of MCTs in SHRSP/Izm and SHRpch1_18 astrocytes. Reduced expression of MCTs may be associated with decreased lactate production in SHRSP.

Morphology of P2X3-immunoreactive nerve endings in the rat laryngeal mucosa.

The morphological characteristics of P2X3-immunoreactive nerve endings in the laryngeal mucosa were herein examined using immunohistochemistry with confocal laser microscopy. Ramified intraepithelial nerve endings immunoreactive to P2X3 were distributed in the epiglottis and arytenoid region. The axon terminals of P2X3-immunoreactive ramified endings were beaded or flat in shape. These endings were also immunoreactive to P2X2 and not identical to the nerve endings immunoreactive to Na(+)-K(+)-ATPase α3-subunit, substance P (SP), and calcitonin gene-related peptide (CGRP). P2X3-immunoreactive axon terminals were also immunoreactive to vGLUT1, vGLUT2, and vGLUT3. In addition to ramified endings, P2X3-immunoreactive nerve endings were associated with α-gustducin-immunoreactive solitary chemosensory cells and/or SNAP25-immunoreactive neuroendocrine cells. Furthermore, P2X3-immunoreactive nerve endings were also observed in the taste bud-like chemosensory cell clusters of the stratified squamous epithelium covering epiglottic and arytenoid cartilage. The P2X3-immunoreactive nerve endings that associated with sensory and/or endocrine cells and chemosensory cell clusters were also immunoreactive to P2X2, vGLUT1, vGLUT2, and vGLUT3, but not to SP or CGRP. In conclusion, P2X3-immunoreactive nerve endings may be classified into two types, i.e., intraepithelial ramified nerve endings and nerve endings associated with chemosensory cells and neuroendocrine cells.

White adipose tissue undergoes browning during preweaning period in association with microbiota formation in mice.

Beige adipocytes are transiently induced during early postnatal period in mice. Previous studies have suggested that, unlike in adults, the induction is independent of the sympathetic nerve activity; however, the mechanism is yet unknown. Here, we showed that beige adipocytes are induced during the preweaning period in association with the formation of microbiota in mice. Alteration of gut microbiota composition in preweaning mice by maternal treatment with antibiotics or high-fat diet feeding substantially suppressed WAT browning. The suppression was also found in pups transplanted cecal microbiota from pups of high-fat diet-fed dams. These treatments reduced the hepatic expression of genes involved in bile acid synthesis and the serum bile acids level. The abundance of Porphyromonadaceae and Ruminococcaceae in microbiota showed a positive and negative correlation with the induction of beige adipocytes, respectively. This finding may provide comprehensive understanding of the association between gut microbiota and adipose tissue development in the neonatal period.

Persistent hyperplastic primary vitreous in two piglets.

Two Large Yorkshire piglets were diagnosed as persistent hyperplastic primary vitreous (PHPV). In case 1, the white cord-like structure extending from optic disc to lens was observed in the normal-sized right eye. Case 2 showed buphthalmos of the right eye. The internal structure of the right eye was unclear due to bleeding, but a white cord-like structure was slightly observed. In both cases, histological examinations revealed the fibrovascular cord-like structure in hyaloid vitreous. The retina was detached, and dysplastic nervous tissue was observed in anterior vitreous. Immunohistochemistry using various neural markers suggested that dysplastic nervous tissue was derived from the detached neural retina. By the characteristic macroscopic and histopathological features, both cases were diagnosed as PHPV. To our knowledge, this is the first report of swine PHPV.

Monovision by Implantation of Posterior Chamber Phakic Intraocular Lens with a Central Hole (Hole ICL) for Early Presbyopia.

This study was aimed to evaluate visual performance at near to far distances in early presbyopic patients undergoing monovision by implantation of an ICL with a central hole (hole ICL). This pilot study comprised thirty-four eyes of 17 early presbyopic patients (age, 40 to 53 years) who underwent hole ICL implantation, and whose targeted refraction was set at emmetropia for the dominant eye, and at slight myopia (-0.5 to -1.0 diopters (D)) for the non-dominant eye. Corrected distance visual acuity was significantly improved, from -0.11 ± 0.07 preoperatively to -0.19 ± 0.09 logMAR postoperatively (p < 0.001, Wilcoxon signed-rank test). Uncorrected distance visual acuity was also significantly improved from 1.43 ± 0.35 preoperatively to -0.04 ± 0.18 logMAR postoperatively (p < 0.001). The mean binocular visual acuity was 0.01 logMAR or better at all distances (5.0, 3.0, 2.0, 1.0, 0.7, 0.5, and 0.3 m). All eyes were within ± 0.5 D of the targeted correction. All patients had within the normal range of near stereopsis. Neither cataract formation, significant intraocular pressure rise, nor vision-threatening complications occurred. Monovision by hole ICL implantation provided good binocular vision at near to far distances, without developing cataract, suggesting its feasibility as a new surgical presbyopic approach for early presbyopia.

Comparison of Size Modulation Standard Automated Perimetry and Conventional Standard Automated Perimetry with a 10-2 Test Program in Glaucoma Patients.

PURPOSE: This prospective observational study compared the performance of size modulation standard automated perimetry with the Octopus 600 10-2 test program, with stimulus size modulation during testing, based on stimulus intensity and conventional standard automated perimetry, with that of the Humphrey 10-2 test program in glaucoma patients. METHODS: Eighty-seven eyes of 87 glaucoma patients underwent size modulation standard automated perimetry with Dynamic strategy and conventional standard automated perimetry using the SITA standard strategy. The main outcome measures were global indices, point-wise threshold, visual defect size and depth, reliability indices, and test duration; these were compared between size modulation standard automated perimetry and conventional standard automated perimetry. RESULTS: Global indices and point-wise threshold values between size modulation standard automated perimetry and conventional standard automated perimetry were moderately to strongly correlated (p < 0.01). However, the correlation coefficient of point-wise threshold value for the central zone was significantly lower than that for the peripheral zone (χ2 > 33.40, p < 0.01). Better mean defect and point-wise threshold values were obtained with size modulation standard automated perimetry than with conventional standard automated perimetry, but the visual-field defect size was smaller (p < 0.01) and depth shallower (p < 0.01) on size modulation-standard automated perimetry than on conventional standard automated perimetry. The reliability indices, particularly the false-negative response, of size modulation standard automated perimetry were worse than those of conventional standard automated perimetry (p < 0.01). The test duration was 6.5% shorter with size modulation standard automated perimetry than with conventional standard automated perimetry (p = 0.02). CONCLUSIONS: Global indices and the point-wise threshold value of the two testing modalities correlated well. However, the potential of a large stimulus presented at an area with a decreased sensitivity with size modulation standard automated perimetry could underestimate the actual threshold in the 10-2 test protocol, as compared with conventional standard automated perimetry.

Diagnostic Ability and Repeatability of a New Supra-Threshold Glaucoma Screening Program in Standard Automated Perimetry.

PURPOSE: We assess the diagnostic ability and repeatability of a new suprathreshold glaucoma screening test (GST) comprising 28 test points and a 1-of-3 sampling strategy at 95% of the normal limit for standard automated perimetry (SAP) in early to advanced glaucoma. METHODS: This prospective cross-sectional study included 96 eyes of patients with early, moderate, or advanced glaucoma and 37 eyes of normal controls. Participants were evaluated by the G-Dynamic threshold test once and the GST twice, in random order, using the Octopus 600 perimeter. The diagnostic ability of GST was assessed by comparison with the G-Dynamic threshold obtained by receiver operating characteristic analysis. Repeatability was assessed by κ statistics for agreement on glaucoma diagnosis and each test point. RESULTS: Although the G-Dynamic test exhibited significantly higher areas under the curve (AUC) than the GST1st (P = 0.009) in early glaucoma, there were no significant differences in any other AUCs between the two methods. The κ values for repeatability of glaucoma diagnosis and each test point were 0.747 to 1.0 and 0.537 to 1.0, respectively. The duration of the GST in the control and early glaucoma groups was less than a minute, while that in the moderate and advanced glaucoma groups was within 1.5 minutes. CONCLUSION: The diagnostic ability of the new suprathreshold GST for early to advanced glaucoma was high, with moderate to strong repeatability and short test duration. TRANSLATIONAL RELEVANCE: There currently are no prominent suprathreshold screening strategies using SAP. The GST would be an effective clinical method for glaucoma screening.

Diagnostic capability of Pulsar perimetry in pre-perimetric and early glaucoma.

This study aimed to compare the diagnostic capability of Pulsar perimetry (Pulsar) in pre-perimetric glaucoma (PPG) and early glaucoma (EG) with that of Flicker perimetry (Flicker) and spectral-domain optical conference tomography (SD-OCT). This prospective cross-sectional study included 25 eyes of 25 PPG patients, 35 eyes of 35 EG patients, and 42 eyes of 42 healthy participants. The diagnostic capability using the area under the curve (AUC) of the best parameter and agreement of detectability between structural and functional measurements were compared. For PPG patients, the AUC of Pulsar, Flicker, OCT-disc, and OCT-macular was 0.733, 0.663, 0.842, and 0.780, respectively. The AUC of Flicker was significantly lower than that of OCT-disc (p = 0.016). For EG patients, the AUC of Pulsar, Flicker, OCT-disc, and OCT-macular were 0.851, 0.869, 0.907, and 0.861, respectively. There was no significant difference in AUC among these methods. The agreement between structural and functional measurements expressed by kappa value ranged from -0.16 to 0.07 for PPG and from 0.01 to 0.25 for EG. Although the diagnostic capability of Pulsar in the PPG and EG groups was equal to that of Flicker and SD-OCT, the agreements between structural and functional measurements for both PPG and EG were poor.

Hypoxia Inducible Factor 1 Alpha Is Expressed in Germ Cells throughout the Murine Life Cycle.

Pluripotent stem cells of the early embryo, and germ line cells, are essential to ensure uncompromised development to adulthood as well as species propagation, respectively. Recently, the transcription factor hypoxia inducible factor 1 alpha (Hif1α) has been shown to have important roles in embryonic stem cells; in particular, regulation of conversion to glycolytic metabolism and, as we have shown, maintenance of functional levels of telomerase. In the present study, we sought to assess whether Hif1α was also expressed in the primitive cells of the murine embryo. We observed expression of Hif1α in pre-implantation embryos, specifically the 2-cell stage, morula, and blastocyst. Robust Hif1α expression was also observed in male and female primordial germ cells. We subsequently assessed whether Hif1α was expressed in adult male and female germ cells. In the testis, Hif1α was robustly expressed in spermatogonial cells, in both juvenile (6-week old) and adult (3-month old) males. In the ovaries, Hif1α was expressed in mature oocytes from adult females, as assessed both in situ and in individual oocytes flushed from super-ovulated females. Analysis of Hif1α transcript levels indicates a mechanism of regulation during early development that involves stockpiling of Hif1α protein in mature oocytes, presumably to provide protection from hypoxic stress until the gene is re-activated at the blastocyst stage. Together, these observations show that Hif1α is expressed throughout the life-cycle, including both the male and female germ line, and point to an important role for Hif1α in early progenitor cells.

Investigation Into the Antinfluenza Agent Oseltamivir Distributed via the Internet in Japan.

This study evaluated the quality and authenticity of the anti-influenza agent oseltamivir acquired via the Internet in Japan. The brand name drugs Tamiflu and Antiflu were obtained via the Internet using a search engine. The authenticity and legitimacy of the medicines were verified with the samples' manufacturers and the regulatory authorities of the country of origin, respectively. In addition, quantitative and dissolution tests were carried out using high-performance liquid chromatography. No counterfeits were detected among the samples. Additionally, all samples passed qualitative and quantitative tests according to the US Pharmacopeia monograph. However, different sorts of mismanagement were observed in the distribution channels. Through the Internet, oseltamivir can be purchased in quantities larger than the permissible amount in Japan. Furthermore, maintenance of medicine quality cannot be guaranteed. In addition, insufficient information and lack of quality control in some samples may cause unwanted health problems in patients.