Enhancing Corneal Drug Penetration Using Penetratin for Ophthalmic Suspensions.

Eye drops, including solutions and suspensions, are essential dosage forms to treat ophthalmic diseases, with poorly water-soluble drugs typically formulated as ophthalmic suspensions. In addition to low bioavailability, suspensions exhibit limited efficacy, safety, and usability due to the presence of drug particles. Improving bioavailability can reduce the drug concentrations and the risk of problems associated with suspended drug particles. However, practical penetration enhancers capable of improving bioavailability remain elusive. Herein, we focused on penetratin (PNT), a cell-penetrating peptide (CPP) that promotes active cellular transport related to macromolecule uptake, such as micropinocytosis. According to the in vitro corneal uptake study using a reconstructed human corneal epithelial tissue model, LabCyte CORNEA-MODEL24, PNT enhanced the uptake of Fluoresbrite® YG carboxylate polystyrene microspheres without covalent binding. In an ex vivo porcine eye model, the addition of 10 µM PNT to rebamipide ophthalmic suspension markedly improved the corneal uptake of rebamipide; however, the addition of 100 µM PNT was ineffective due to potentially increased particle size by aggregation. This article provides basic information on the application of PNT as a penetration enhancer in ophthalmic suspensions, including the in vitro and ex vivo studies mentioned above, as well as the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cytotoxicity assay and storage stability at different pH values.

Exploring Cell-Penetrating Peptides as Penetration Enhancers in Eye Drop Formulations Using a Reconstructed Human Corneal Epithelial Model.

Ocular tissues function as biological barriers that hinder drug delivery, depending on the target tissue and route of administration, and must be overcome to achieve the desired therapeutic effect. Penetration enhancers have long been investigated to improve corneal drug penetration via eye drop instillation; however, further development is warranted owing to potential safety concerns. In the present study, we focused on cell-penetrating peptides (CPPs) as a penetration enhancer to address the requirements and explored CPP candidates suitable for corneal drug delivery. Using a reconstructed human corneal epithelial tissue model, LabCyte CORNEA-MODEL24 as an alternative to animal testing that is expected to have higher reproducibility than extracted eyeballs and octa-arginine (R8) as a representative model CPP with simple structure, we investigated the enhancement of 6-carboxyfluorescein (6-FAM) uptake by fluorescence imaging and the potential of eye irritation by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Also, surface plasmon resonance (SPR) evaluated the interaction between R8 and model compounds, suggesting that the stronger interaction could facilitate the corneal uptake of compounds. A comparative screening study of corneal uptake using various CPPs showed that the CPPs other than R8 also have the potential to enhance the corneal uptake of 6-FAM. In particular, penetratin (PNT) showed stronger fluorescence intensity. Through these findings, this manuscript provides beneficial information for the development of a novel corneal penetration enhancer with CPPs. In the future, it is expected that the basic findings with R8 will be verified to be applicable to other CPPs for development as penetration enhancers for eye drop formulation.

Beneficial effects of combination therapy of canagliflozin and teneligliptin on diabetic polyneuropathy and ß-cell volume density in spontaneously type 2 diabetic Goto-Kakizaki rats.

AIMS: Parasympathetic nerve (PN) signaling plays a crucial role in the maintenance of pancreatic ß-cell volume density (Vß). PN may be pathologically affected in diabetic polyneuropathy (DPN). However, the association between the reduction of PNs in islets and Vß and the therapeutic effects of a DPP4 inhibitor (DPP4i) and an SGLT2 inhibitor (SGLT2i) in nonobese type 2 diabetes mellitus (T2DM) Goto-Kakizaki rats (GK) have not been investigated. MATERIALS AND METHODS: We divided 5-week old male GK and Wistar rats (W) into a DPP4i-treated group (GKTe), SGLT2i-treated group (GKCa), and combination-treated group (GKCaTe). After 25 weeks, the pancreata was pathologically evaluated. RESULTS: Vß in GK was significantly decreased (p < 0.01 vs. W), whereas Vß was the most well preserved in GKCaTe (p < 0.05 vs. GKTe), followed by GKTe (p < 0.05 vs. GK). The decreased amount of PNs in the islets and intraepidermal nerve fiber density (IENFD) in GK was significantly improved in the treated groups compared with GK (p < 0.05 vs. GKCa and GKTe and p < 0.01 vs. GKCaTe). PN density and IENFD were significantly correlated with Vß (r = 0.55, p < 0.01 and r = 0.54, p < 0.01, respectively). IENFD was identified as a surrogate marker for the prediction of Vß (cutoff value, 16.39). CONCLUSIONS: The combination therapy of DPP4i and SGLT2i improved Vß accompanied by PNs density and IENFD. IENFD was proportionally correlated with Vß. Therefore, the prevention of DPN development may be concurrently beneficial for the preservation of Vß in nonobese T2DM.

Corrigendum: Normal High HbA1c a Risk Factor for Abnormal Pain Threshold in the Japanese Population.

[This corrects the article DOI: 10.3389/fendo.2019.00651.].

Diabetes, an independent poor prognostic factor of non-B non-C hepatocellular carcinoma, correlates with dihydropyrimidinase-like 3 promoter methylation.

A concurrent increase in the prevalence of hepatocellular carcinoma (HCC) with that of type 2 diabetes (T2D) and obesity has been reported in the absence of hepatitis B virus surface antigen-negative/hepatitis C virus antibody-negative HCC (NBNC-HCC). However, the prognostic relevance of this association remains unclear. Promoter methylation (PM) of the dihydropyrimidinase-like 3 gene (DPYSL3) has been implicated in virus-related HCC. However, it remains unclear whether T2D influences PM in NBNC-HCC. We determined the influence of T2D on clinicopathological profile and PM of DPYSL3 and CDK2NA in patients with NBNC-HCC who were divided into two groups: non-diabetes (non-DM; n = 46) and diabetes (DM; n = 47). DM was associated with a higher Union for International Cancer Control grade, marginal vascular invasion and tumour cell proliferation irrespective of the duration of T2D as well as higher rates of PM of DPYSL3 than non-DM; however, PM of CDK2NA was similar between both groups. PM of DPYSL3 reduced its expression which inversely correlated with reduced patient survival. In conclusion, T2D is associated with poor prognosis of NBNC-HCC in which a high frequency of PM of DPYSL3 may play a pivotal role in its pathogenesis.

Normal High HbA1c a Risk Factor for Abnormal Pain Threshold in the Japanese Population.

Purpose: Small fiber dysfunction is common in subjects with diabetic polyneuropathy (DPN). It is unsettled, however, whether marginal glucose intolerance is implicated in the onset and progression of small fiber dysfunction. Herein, we explored the relationship between glycated hemoglobin levels (HbA1c) and pain sensation in the Japanese population. Methods: A population-based study of 894 individuals (352 men, 542 women; average age 53.8 ± 0.5 years) and 55 subjects with impaired fasting glucose (IFG) in the 2017 Iwaki project were enrolled in this study. Individuals with diabetes were excluded. Relationships between pain threshold for intraepidermal electrical stimulation (P-IES) and parameters associated with metabolic syndrome were examined. Results: P-IES was elevated with increasing of age in women but not in men. Average P-IES (mA) was increased in IFG subjects (n = 55, 0.20 ± 0.03) compared with normoglycemic/non-IFG individuals (n = 894, 0.15 ± 0.11) (p < 0.01). It was comparable between IFG and a group of normal high HbA1c (5.9-6.4%). Univariate linear regression analyses showed no influence of sex, triglyceride, or cholesterol on the value of P-IES. In contrast, there were significant correlations between P-IES and serum HbA1c level (ß = 0.120, p < 0.001) Adjustments for the multiple clinical measurements confirmed positive correlation of P-IES with HbA1c (ß = 0.077, p = 0.046). Conclusion: Individuals with normal high HbA1c exhibited an elevated P-IES in a healthy Japanese population which may be useful for the screening of subclinical DPN.

Worsened outcome in patients with pancreatic ductal carcinoma on long-term diabetes: association with E-cadherin1 (CDH1) promoter methylation.

Prevalence of pancreatic ductal carcinoma (PDC) is nearly twice in patients with diabetes mellitus, but the reason for this close association remains obscure. Recently promoter methylation of E-cadherin1 (CDH1) and CDKN2A genes, encoding E-cadherin and P16 respectively, are invoked in development of PDC. It is still unclear whether diabetes affects such epigenetic changes and malignant behavior in PDC. In this study, we studied whether diabetes influences the clinico-pathological profile and methylation status of CDH1 and CDKN2A genes in patients with PDC. PDC subjects were divided into 3 groups; 59 cases without diabetes (non-DM), 17 cases with short-term diabetes (short-DM)(diabetes duration 3 yrs>) and 33 cases with long-term diabetes (long-DM)(≧3 yrs). Compared to non-DM or short-DM, long-DM was associated with a higher histological grade of malignancy and a higher tumor stage. Promoter methylation of both CDH1 and CDKN2A was encountered more frequently in PDC patients with long-DM than non-DM or short DM. Cases with CDH1 promoter methylation showed reduced E-cadherin expression and worsened survival. We consider that the presence of long-DM has a negative impact on the prognosis of PDC patients which may be relevant to a high frequency of promoter methylation of CDH1.

Thermal Molding of Organic Thin-Film Transistor Arrays on Curved Surfaces.

In this work, a thermal molding technique is proposed for the fabrication of plastic electronics on curved surfaces, enabling the preparation of plastic films with freely designed shapes. The induced strain distribution observed in poly(ethylene naphthalate) films when planar sheets were deformed into hemispherical surfaces clearly indicated that natural thermal contraction played an important role in the formation of the curved surface. A fingertip-shaped organic thin-film transistor array molded from a real human finger was fabricated, and slight deformation induced by touching an object was detected from the drain current response. This type of device will lead to the development of robot fingers equipped with a sensitive tactile sense for precision work such as palpation or surgery.

Physical Property Evaluation of ZnO Thin Film Fabricated by Low-Temperature Process for Flexible Transparent TFT.

The usual silicon-based display back planes require fairly high process temperature and thus the development of a low temperature process is needed on flexible plastic substrates. A new type of flexible organic light emitting transistor (OLET) had been proposed and investigated in the previous work. By using ultraviolet/ozone (UV/O3) assisted thermal treatments on wet processed zinc oxide field effect transistor (ZnO-FET), through low-process temperature, ZnO-FETs were fabricated which succeeded to achieve target drain current value and mobility. In this study, physical property evaluation of ZnO was conducted in term of their crystallinity, the increase composition of ZnO formed inside the thin film and the decrease of the carbon impurities originated from aqueous solution of the ZnO itself. The X-ray diffraction (XRD) evaluation showed UV/03 assisted thermal treatment has no obvious effect towards crystallinity of ZnO in the range of low process temperature. Moreover, through X-ray photoelectron spectroscopy (XPS) evaluation and Fourier transform infrared (FT-IR) spectroscopy evaluation, more carbon impurities disappeared from the ZnO thin film and the increase of composition amount of ZnO, when the thin film was subjected to UV/O3 assisted thermal treatment. Therefore, UV/O3 assisted thermal treatment contributed in carbon impurities elimination and accelerate ZnO formation in ZnO thin film, which led to the improvement in the electrical property of ZnO-FET in the low-process temperature.

Partial Dissolution of Charge Order Phase Observed in ß-(BEDT-TTF)2PF6 Single Crystal Field Effect Transistor.

The effect of an applied gate electric field on the charge-order phase in ß-(BEDT-TTF)2PF6 single-crystal field-effect transistor structure was observed at around room temperature by technical improvement with respect to sample preparation and electrical measurements. A relatively slight but systematic increase of the electrical conductance induced by the applied gate electric field and its temperature dependence was observed at around the metal-insulator transition temperature (TMI). The temperature dependence of the modulated electrical conductance demonstrated that TMI was shifted toward the lower side by application of a gate electric field, which corresponds to partial dissolution of the charge-order phase. The thickness of the partially dissolved charge order region was estimated to be several score times larger than the charge accumulation region.

Anticancer effects of fucoxanthin and fucoxanthinol on colorectal cancer cell lines and colorectal cancer tissues.

Colorectal cancer is one of the most malignant neoplasms worldwide. Fucoxanthin is a carotenoid present in the chloroplasts of brown seaweeds. In the present study, the anticancer effects of fucoxanthin and its metabolite, fucoxanthinol, on 6 colorectal cancer cell lines and 20 tissue samples from surgically resected clinical colorectal cancer specimens were examined using a collagen-gel droplet embedded culture drug sensitivity test (CD-DST). The in vitro sensitivity to fucoxanthin, fucoxanthinol and the anticancer drugs is expressed as T/C (%), where T is the absorbance of cells which stained by neutral red treated with carotenoids and C is the absorbance of non-staining cells. Fucoxanthin and fucoxanthinol decreased the T/C (%) of Caco-2, WiDr, HCT116, and DLD-1 cell lines at doses of 20 µM. Fucoxanthinol also decreased the T/C (%) of SW620 cells, while the T/C (%) of Colo205 cells was not reduced by treatment with either carotenoid. Specifically, the T/C (%) of Caco-2 and WiDr cells, which were incubated in carotenoid-free medium for 6 days following treatment with 20 µM fucoxanthinol for 24 h, was markedly decreased to 1.4±0.2 and 12.0±0.3%, respectively. Furthermore, fucoxanthin and fucoxanthinol decreased the T/C (%) in colorectal cancer tissue samples. Notably, 20 µM fucoxanthinol treatment resulted in a higher proportion of colorectal cancer samples with a T/C (%) of <50% (13/20, 65%) compared with samples treated with 20 µM fucoxanthin (2/20, 10%). The median T/C (%) value of 35.1% for the 20 cancers specimens treated with 20 µM fucoxanthinol was lower than the median T/C (%) values of 86.3% and 75.8% for those treated with fluorouracil and paclitaxel, respectively. These results suggested that fucoxanthin and fucoxanthinol may be of use as chemotherapeutic agents in colorectal cancer.

Gap states in pentacene thin film induced by inert gas exposure.

We studied gas-exposure effects on pentacene (Pn) films on SiO2 and Au(111) substrates by ultrahigh sensitivity photoelectron spectroscopy, which can detect the density of states of ∼10(16) states eV-1 cm-3 comparable to electrical measurements. The results show the striking effects for Pn/SiO2: exposure to inert gas (N2 and Ar) produces a sharp rise in gap states from ∼10(16) to ∼10(18) states eV-1 cm-3 and pushes the Fermi level closer to the valence band (0.15-0.17 eV), as does exposure to O2 (0.20 eV), while no such gas-exposure effect is observed for Pn/Au(111). The results demonstrate that these gap states originate from small imperfections in the Pn packing structure, which are induced by gas penetration into the film through the crystal grain boundaries.

HCV virus and lymphoid neoplasms.

Hepatitis C virus (HCV) is one of the viruses known to cause hepatic cancer. HCV is also believed to be involved in malignant lymphoma. In this paper, we investigated characteristics of malignant lymphoma cases that were anti-HCV antibody (HCV-Ab) positive. We were able to perform pathological examinations on 13 out of 14 HCV-positive cases. Of these, lymphoid tissues of 10 stained positive for HCV-Ab. There was no significant correlation between the degree of HCV staining and the rate of recurrence or resistance to treatment. However, there did appear to be a consistent decrease in the amount of HCV-RNA between pre- and posttreatment among HCV-Ab-positive cases; that is, treatment-resistant cases that exhibited resistance from the first treatment and recurrent cases more frequently had a higher HCV level at treatment termination compared to the pretreatment level. This suggests that the HCV virus either accelerates oncogenesis by direct interaction with B cells or indirectly affects lymphoma prognosis.

Changes in the fatty acid composition of the liver with the administration of N-3 polyunsaturated fatty acids and the effects on warm ischemia/reperfusion injury in the rat liver.

Prostanoids play a pivotal role among the inflammatory mediators associated with I/R injury. The aim of this study was to determine the effects of oral supplementation of n-3 polyunsaturated fatty acids (PUFA)-rich oil on inflammatory reactions and microcirculatory disorders caused by a hepatic warm I/R in rats. The rats were orally supplemented with n-3 PUFA-rich oil, n-6 PUFA-rich oil, or the same volume of water for 7 days. The PUFA concentration in the blood and liver tissues were evaluated, and the effects on I/R injury of the liver were assessed. The n-3 PUFA supplementation elevated the n-3/n-6 ratio in the blood and liver tissues. After reperfusion, thromboxane B(2) in the blood and prostaglandin E(2) in the liver were significantly suppressed in the n-3 PUFA-treated rats. Hepatic microcirculation was well maintained from the early phase (30 min) of reperfusion, and the serum concentrations of TNF-alpha and IL-6 were significantly lower in this group. The transaminase blood levels were also suppressed in the n-3 PUFA-treated rats. Expression of COX-2 mRNA was increased in all groups at 2 h after reperfusion but there were no differences among three groups. In conclusion, preoperative n-3/n-6 ratio augmentation in the blood and in the liver can result in a successful alleviation of hepatic I/R injury.

Domino double Michael-Claisen cyclizations: a powerful general tool for introducing quaternary stereocenters at C4 of cyclohexane-1,3-diones and total synthesis of diverse families of sterically congested alkaloids.

Reactions of substituted acetone derivatives with acrylic acid esters (>200 mol %) in the presence of t-BuOK (200 mol %) in t-BuOH-THF (1:1 by volume) turned out to proceed as a cascade process consisting of the first Michael addition, the second Michael addition, and the last Claisen reaction to afford 4,4-disubstituted cyclohexane-1,3-diones. Only more substituted enolates play the role of a Michael donor in this cascade process, and therefore the ketone took up two alkoxycarbonylethyl groups on the same carbon bearing more substituents. Such intermediates were followed by intramolecular Claisen reactions leading to cyclohexane-1,3-diones bearing quaternary stereogenic centers at C(4), which bears an alkoxycarbonylethyl group and the substituent of the starting acetone derivatives. Thus-obtained 4,4-disubstituted cyclohexane-1,3-diones were successfully employed for total syntheses of intricate alkaloids of biological interest such as (+)-aspidospermidine, (+/-)-galanthamine, (+/-)-lycoramine, and (+/-)-mesembrine, all featuring quaternary stereogenic centers. DFT calculations provided us with clear-cut explanations for the observed chemoselectivity of the cascade process involving ketone-based enolates under thermodynamically controlled conditions.

HCV non-structural protein 3 and HCV RNA genome in non-Hodgkin lymphoma and transition of the serum HCV RNA level: a retrospective analysis in one institution.

There have been various reports on the association of hepatitis C virus (HCV) infection with B lymphocyte proliferative disorders, such as non-Hodgkin lymphoma (NHL). We experienced a case (Case 1) of anti-HCV antibody (HCV-Ab)-positive NHL in which HCV nonstructural protein 3 (NS3) expression was observed in lymphoma tissue at the time of recurrence and in which the serum HCV RNA level exhibited a transient increase prior to recurrence. We investigated the HCV RNA genome in lymphoma tissue in Case 1, and it could be detected at recurrence. We also investigated HCV NS3 protein expression in lymphoma tissue and changes in serum HCV RNA level during the clinical course in four other cases of HCV-Ab-positive NHL treated in our hospital. We examined lymphoma tissues for HCV NS3 protein expression in four of the five cases, but it was not identified except for in Case 1 at recurrence. In three cases with no recurrence, serum HCV RNA levels showed a tendency to decrease after completion of chemotherapy and became stable thereafter. Further studies are necessary to clarify the association between serum HCV RNA and the onset and exacerbation of NHL.

Small cell undifferentiated carcinoma of the ascending colon with rapid enlargement after resection: report of a case and review of the literature.

Although adenocarcinoma represents the vast majority of neoplasms of the large intestine, small cell undifferentiated carcinoma (SCUC) also arises from the colorectum. SCUC of the colorectum is highly malignant and shares the similarities in histologic characteristics, behavior, and histochemistry with SCUC of the lung. We report herein a case of SCUC in the ascending colon with rapid enlargement after resection. A 70-year-old male, who presented to a nearby physician with chief complaints of pain in the right lower quadrant, was referred to our hospital. We found a tumor mass that was approximately 15 cm in size in the right lower quadrant. Computed tomography (CT) images showed an irregularly shaped tumor, located inferior to the lower border of the right kidney and in the area of the ascending colon. By colonoscopy, we found a circumferential tumor of the ascending colon. A biopsy indicated it to be SCUC. The patient underwent right hemicolectomy. Two weeks after the resection, we palpated a tumor mass in the same area. The abdominal CT images showed a tumor mass that was approximately 10 cm in size. The tumor rapidly enlarged, and the patient died of multiple organ failure. SCUC is a tumor with a high malignant potential. Radical treatment cannot be achieved by surgical therapy alone and hence further studies of effective adjuvant therapy would be required.

Efficacy of training program for ambulatory competence in elderly women.

The optimal prevention of osteoporotic fractures in the elderly consists of increasing the bone density and preventing falls. We report on the efficacy of training program to promote ambulatory competence in elderly women. Twenty-five elderly women were enrolled in our training program, which is a three-month program consisting of dynamic balance training with Galileo 900 (Novotec, Pforzheim, Germany) once a week, combined with daily static balance (standing on one leg like a flamingo) and resistance (half-squat) training. The mean age of the participants was 72.8 years (range, 61-86 years). After 3 months of training, the step length, knee extensor muscle strength, and maximum standing time on one leg were significantly increased, while the walking speed and hip flexor muscle strength were not significantly altered. During the study period, no serious adverse events such as new vertebral fractures or adverse cardiovascular symptoms were observed in any participant. The present preliminary study shows that our training program may have the potential to promote ambulatory competence in elderly women.