Galectin-3, a damage-associated molecular pattern, in tears of patients with vernal keratoconjunctivitis.

PURPOSE: Galectin-3 is a damage-associated molecular pattern (DAMPs), released from damaged or dying cells. In this study, we investigated the concentration and source of galectin-3 in the tears of patients with vernal keratoconjunctivitis (VKC) and evaluated whether the concentration of galectin-3 in tears represents a biomarker of corneal epithelial damage. STUDY DESIGN: Clinical and experimental. METHODS: We measured the concentration of galectin-3 in tear samples from 26 patients with VKC and 6 healthy controls by enzyme-linked immunosorbent assay (ELISA). The expression of galectin-3 in cultured human corneal epithelial cells (HCEs) stimulated with or without tryptase or chymase was investigated by polymerase chain reaction (PCR), ELISA, and Western blotting. We also estimated the concentration of galectin-3 in the supernatants of cultured HCEs induced to necrosis. Finally, we investigated whether recombinant galectin-3 induced the expression of various genes related to cell migration or the cell cycle in HCEs by using microarray analysis. RESULTS: High concentrations of galectin-3 were detected in the tears of patients with VKC. The concentration showed significant correlation with the severity of corneal epithelial damage. Stimulation of cultured HCEs with various concentrations of tryptase or chymase had no effect on the expression of galectin-3. However, high concentrations of galectin-3 were detected in the supernatants of necrotic HCEs. Recombinant human galectin-3 induced various cell migration- and cell cycle-related genes. CONCLUSION: The concentrations of galectin-3 in the tears of patients with VKC may represent a biomarker of the severity of corneal epithelial damage.

Role of oncostatin M in the pathogenesis of vernal keratoconjunctivitis: focus on tissue remodeling.

PURPOSE: Vernal keratoconjunctivitis (VKC) is a severe and recurrent allergic conjunctivitis, the mechanism of which is not well understood. In this study, we investigated the role of oncostatin M (OSM) in the pathogenesis of VKC, with a focus on tissue remodeling. STUDY DESIGN: Clinical and experimental. PATIENTS AND METHODS: The OSM concentrations in tear fluid samples obtained from VKC patients and healthy controls were measured using ELISA, and the expression of OSM mRNA and protein in giant papillae resected from VKC patients was investigated using RT-PCR and immunohistochemistry, respectively. In cultured human conjunctival epithelial cells (HconEpiCs), expression of OSM receptor ß (OSMRß) was detected using immunocytochemical and FACS analyses. Finally, we investigated whether recombinant OSM activated STAT1 and STAT3 to induce the expression of various genes related to tissue remodeling in HconEpiCs, by using Western blot analysis, microarray analysis, and RT-PCR. RESULTS: The OSM concentration was higher in the tear fluid of VKC patients than in that of the healthy controls, and strong expression of OSM mRNA was found in the giant papillae. We also detected T cells expressing OSM in the giant papillae. In addition, HconEpiCs showed surface expression of OSMRß. Recombinant human OSM strongly activated both STAT1 and STAT3 in HconEpiCs and induced various tissue remodeling-related genes, including MMP-1, MMP-3, IL-24, IL-20, serpinB3, S100A7, tenascin C, and SOCS3. CONCLUSION: Our results suggest that OSM is one of the key molecules involved in remodeling of giant papillae in VKC.

Pulmonary liposomal formulations encapsulated procaterol hydrochloride by a remote loading method achieve sustained release and extended pharmacological effects.

Drug inhalation provides localized drug therapy for respiratory diseases. However, the therapeutic efficacy of inhaled drugs is limited by rapid clearance from the lungs. Small hydrophilic compounds have short half-lives to systemic absorption. We developed a liposomal formulation as a sustained-release strategy for pulmonary delivery of procaterol hydrochloride (PRO), a short-acting pulmonary ß2-agonist for asthma treatment. After PRO-loaded liposomes were prepared using a pH gradient (remote loading) method, 100-nm liposomes improved residence times of PRO in the lungs. PRO encapsulation efficiency and release profiles were examined by screening several liposomal formulations of lipid, cholesterol, and inner phase. Although PRO loading was not achieved using the conventional hydration method, PRO encapsulation efficiency was >60% using the pH gradient method. PRO release from liposomes was sustained for several hours depending on liposomal composition. The liposomal formulation effects on the PRO behavior in rat lungs were evaluated following pulmonary administration in vivo. Sustained PRO release was achieved using simplified egg phosphatidylcholine (EPC)/cholesterol (8/1) liposome in vitro, and greater PRO remnants were observed in rat lungs following pulmonary administration. Extended pharmacological PRO effects were observed for 120min in a histamine-induced bronchoconstriction guinea pig model. We indicated the simplified EPC/cholesterol liposome potential as a controlled-release PRO carrier for pulmonary administration.

Characterization of a doxorubicin liposome formulation by a novel in vitro release test methodology using column-switching high-performance liquid chromatography.

A novel in vitro release test methodology for a liposome formulation was developed using a column-switching high-performance liquid chromatography (HPLC) system. Doxorubicin (DXR) liposome formulations were used as a model. A DXR liposome formulation was dispersed into a release medium, and the dispersion fluid was directly injected at predetermined time points into the column-switching HPLC system. To evaluate the release profile, this system can be used for determining the released and encapsulated DXR in the liposome formulation separately. Comparison with a conventional in vitro release test methodology by dialysis revealed that the methodology developed by column-switching HPLC had no rate-limiting process of membrane permeation of the drug (which is occasionally observed in the dialysis method). The in vitro release profiles of DXR liposome formulations were well characterized using the method developed by column-switching HPLC, and different in vitro release characteristics were revealed. The developed method did not require a large amount of sample or a complicated pretreatment. In addition, the developed column-switching HPLC system was applicable for characterization of the encapsulation profile of liposome formulations.

Identification of five phytosterols from Aloe vera gel as anti-diabetic compounds.

The genus Aloe in the family Liliaceae is a group of plants including Aloe vera (Aloe barbadensis MILLER) and Aloe arborescens (Aloe arborescens MILLER var. natalensis BERGER) that are empirically known to have various medical efficacies. In the present study, we evaluated the anti-hyperglycemic effect of Aloe vera gel and isolated a number of compounds from the gel. On the basis of spectroscopic data, these compounds were identified as lophenol, 24-methyl-lophenol, 24-ethyl-lophenol, cycloartanol, and 24-methylene-cycloartanol. These five phytosterols were evaluated for their anti-hyperglycemic effects in type 2 diabetic BKS.Cg-m(+/+)Lepr(db/J) (db/db) mice. In comparison with the hemoglobin A1c (HbA1c) levels of vehicle-treated mice, statistically significant decreases of 15 to 18% in HbA1c levels were observed in mice treated with 1 mug of the five phytosterols. Considering the ability to reduce blood glucose in vivo, there were no differences between the five phytosterols. Administration of beta-sitosterol did not reduce the blood glucose levels in db/db mice. After administration of the five phytosterols for 28 d, fasting blood glucose levels decreased to approximately 64%, 28%, 47%, 51%, and 55% of control levels, respectively. Severe diabetic mice treated with phytosterols derived from Aloe vera gel did not suffer weight reduction due to glucose loss in the urine. These findings suggest that Aloe vera gel and phytosterols derived from Aloe vera gel have a long-term blood glucose level control effect and would be useful for the treatment of type 2 diabetes mellitus.

[Perioperative changes in the circulatory blood volumes in patients undergoing the abdominal aortic aneurysm repair].

BACKGROUND: The effect of the intraoperative volume load on the circulating blood volume is unclear. Therefore, we investigated the changes of the perioperative blood volume in patients undergoing major surgery. METHODS: In 16 patients undergoing abdominal aortic aneurysm repair, we measured the circulating blood volume (BV x kg(-1)) with the pulse dyedensitometry method (DDG analyzer) before operation, just after operation, on the first postoperative day (1POD) and 2POD. Anesthesia was maintained with general anesthesia and epidural analgesia. The volume of infusion depended on empiric judgment by an anesthesiologist. RESULTS: Mean anesthesia time was 390 minutes, the mean volume of infusion 4225 ml, transfusion 1329 g, blood loss 1567 g, urine output 729 ml, and mean water balance +3287 ml. The mean body weight (BW) was 59.2 +/- 13.7 (before), 60.2 +/- 14.4 (just after), 60.5 +/- 14.3 (1POD), and 60.3 +/- 14.3 (2POD) kg, respectively. The mean BV x kg(-1) was 78.0 +/- 19.0 (before), 65.6 +/- 13.4 (just after), 70.4 +/- 15.7 (1POD), and 76.5 +/- 18.2 (2POD) ml x kg(-1), respectively. The mean BW after operation increased significantly from the value before surgery. However, the mean BV x kg(-1) decreased significantly. There was a significant correlation between the intraoperative increase in BW and intraanesthetic water balance (Y = -2.91 + 0.071X; R = 0.66, P = 0.0412), but there was no significant correlation between the change in BV x kg(-1) and intraanesthetic water balance. CONCLUSIONS: Intraoperative water balance influenced a change of BW significantly, but did not affect BV x kg(-1).

[Anesthetic management of caesarean section in a patient with asymptomatic idiopathic prolonged QT interval syndrome].

A 27-year-old patient with asymptomatic idiopathic prolonged QT interval syndrome underwent elective caesarean section. We selected combined spinal-epidural anesthesia to avoid excessive sympathetic stimulation. Bupivacaine 9 mg and lidocaine 100 mg were used for spinal anesthesia and epidural anesthesia, respectively. The anesthetic course was uneventful, and the infant showed no complication during the perioperative period. In conclusion, we consider combined spinal-epidural anesthesia as a safe anesthetic strategy for caesarean section in a patient with asymptomatic idiopathic prolonged QT interval syndrome.

[A case of Churg-Strauss syndrome undergoing cesarean section].

A 37-year-old woman with Churg-Strauss syndrome underwent cesarean section under combined spinal-epidural anesthesia. Churg-Strauss syndrome is a rare diffuse vasculitis accompanied by severe asthma. Anesthesia was performed uneventfully, but there were several issues of concern regarding the perioperative management of this syndrome.