[Evaluation of tuberculosis treatment including levofloxacin (LVFX) in cases who could not continue standard regimen].

OBJECTIVE: The purpose of this study was to evaluate tuberculosis treatment including levofloxacin (LVFX) and to investigate the effectiveness of changing drug regimens at our hospital. SUBJECTS AND METHODS: A retrospective study was conducted on 331 patients with tuberculosis admitted to Tokyo National Hospital in 2005. Out of these 331 patients, LVFX was used in 48 (14.5%), 41 of which were initial-treatment cases. We studied why and how LVFX was used and compared bacteriological negative conversion rates between the initial-treatment cases in which the initial standard regimen was changed to regimens including LVFX, and those in which the initial standard regimen was either maintained throughout or modified with drugs other than LVFX. Sputum cultures were examined with Mycobacteria Growth Indicator Tube System (BACTEC MGIT 960). RESULTS: LVFX was used in 41 (13.6%) of 302 initial-treatment cases and in 7 (24.1%) of 29 retreatment cases. Out of the 269 initial-treatment cases starting with the standard regimen, LVFX was later used in 26 cases (9.7%). The reasons for using LVFX were adverse reaction to antituberculosis drugs in 23 cases (88.5%) and resistance to antituberculosis drugs in 3 cases (11.5%). We investigated the bacteriological conversion rate in 228 patients who could be followed up for more than five months. The conversion rates in 105 cases under the standard regimen including PZA (PZA+) were 92.4% in three months, 98.1% in four months, and 100% in five months. The rates in 56 cases under the standard regimen without PZA (PZA-) were 92.9 %, 98.2% and 100%,respectively. The rates of 22 cases under the initial regimen modified with LVFX (LVFX +) were 68.2 %, 95.5% and 100%, respectively. In 45 cases under the initial regimen modified with drugs other than LVFX (LVFX-), the rates were 80.0%, 97.8% and 100%, respectively. CONCLUSION: This study showed that LVFX was an effective drug in terms of the bacteriological conversion rate, without adverse reaction. LVFX is not approved as an antituberculosis drug in Japan, but it is often used in cases of MDR-TB or in situations in which the patients cannot continue treatment with the standard regimen. We hope that LVFX will be approved as an antituberculosis drug as soon as possible in Japan.

Histochemical localization of neutral proteases released during development of rat periradicular lesion.

OBJECTIVE: The purpose of the present study was to examine the role of various neutral proteases released during the development of periradicular lesion. DESIGN: This lesion produced by pulpal exposure of mandibular first molar in rat. The histological and histometrical changes in periapical tissue examined. The presence of neutrophil elastase, cathepsin G, collagenase 2, gelatinase B, and secretory leukocyte protease inhibitor (SLPI) was immunohistochemically evaluated in the periapical tissue. RESULTS: After pulpal exposure, some inflammatory cells were present in the periapical tissue at 7 days, and periapical inflammation gradually increased. Alveolar bone resorption observed after 14 days and apical abscess found after 21 days. After 14 days, the area of periradicular lesion significantly increased compared from normal one (p<0.05). Neutrophil elastase, cathepsin G, collagenase 2, and gelatinase B were detected around the root apex at 14 days, then these proteases increased gradually and demonstrated in and around the abscess at 21 and 28 days. Morphologically, these protease-expressing cells are almost polymorphic and polynuclear shaped after 14 days. These cells significantly increased after 14 days compared from normal one (p<0.05). However, SLPI detected after 21 days around apical root. It significantly increased after 21 days (p<0.05). CONCLUSIONS: These results suggested that neutrophil elastase, cathepsin G, collagenase 2, and gelatinase B induce the destruction of periapical tissue. We demonstrated that these neutral proteases released play an important role in development of periradicular lesion.

Morphological observations of rat corneal endothelial cells after exposure to ozonated solution.

PURPOSE: To determine whether exposure to ozonated solution alters the morphology of corneal endothelial cells in rats and to examine the protective effect of ascorbic acid. METHODS: The anterior chambers of rat eyes were filled with 4 ppm of ozonated solution. Some were left in that state, while others were flushed out either 10, 30, or 60 s after exposure to a balanced salt solution (BSS), or to BSS containing 0.001 M ascorbic acid. Corneal endothelial cells were assessed by scanning and electron microscopy either 1 h or 1 week after treatment, and the expressions of aquaporin (AQ)-1 and zonula occludens (ZO)-1 were determined by immunohistochemistry. RESULTS: When exposure time was longer than 10 s, damaged cell membranes and abnormal organelles were observed 1 h after treatment. The longer the exposure time, the more severe the observed alterations; however, the eyes regained almost their normal state at 1 week. When the BSS contained ascorbic acid, no severe damage was observed under any condition. Normal AQ-1 and ZO-1 expressions were observed even with 60 s of exposure when ascorbic acid was used. CONCLUSIONS: A short period of irrigation of the anterior chamber with ozonated solution does not harm the corneal endothelium even when used in combination with ascorbic acid.

Increases in central retinal artery blood flow in humans following carotid artery and stellate ganglion irradiation with 0.6 to 1.6 microm irradiation.

The authors applied near-infrared low-level laser irradiation (LLLI) directed to the stellate ganglion (SG) and to the common carotid artery (CCA), and compared the effects on central retinal artery blood flow using color pulse Doppler sonography. In 10 healthy volunteers, LLLI (0.92 W, 1 : 1 duty cycle, 10 min) to both the SG and CCA significantly increased peak systolic blood velocity in the ophthalmic artery (p<0.001, each) and central retinal artery (p<0.001, each) without changes in vessel resistance. Irradiation to the CCA produced a stronger effect than that to the SG in the ophthalmic artery (p=0.007) and central retinal artery (p=0.031). These data suggest that LLLI to the SG or to the CCA is a useful therapy for increasing the retrobulbar blood flow, with irradiation directed to the CCA being more effective than that directed to the SG in clinical settings.

Quantitative analysis of posterior capsule opacification of hydrophobic acrylic intraocular lenses.

Posterior capsule opacification (PCO) remains a common complication of modern cataract surgery, although both modification of materials used and changes in the intraocular lens (IOL) optic edge design have helped to decrease its incidence slightly. Recently, various kinds of quantitative methods have been developed for measuring PCO. The purpose of this study was to compare the quantitative analysis of PCO between different types of IOL designs. Patients enrolled in the study had age-related cataract and underwent uneventful cataract surgery and implantation of either the AcrySof MA30BA (Alcon) or the Sensor AR40e (AMO), which are differently designed hydrophobic acrylic IOLs with a sharp-edged optic design. Postoperative examination was performed at 6 months. Retroillumination photographs of each eye were obtained, and the degree of PCO was assessed using the Evaluation of Posterior Capsule Opacification (EPCO) system. Grade 1 PCO was noted in both the MA30BA and the AR40e groups. There was no significant difference in the mean PCO score between the MA30BA and AR40e groups. Although the sharp-edged optic designs of both IOLs might similarly inhibit PCO at 6 months, a long-term follow-up period is needed to determine if any PCO differences occur between these 2 hydrophobic acrylic IOLs.

Two cases of true exfoliation of the lens capsule after cataract surgery.

True exfoliation of the lens capsule is known to be associated with glassblower's cataract, which is caused by extended exposure to excessive heat. Furthermore, inflammation and trauma are also considered to be predisposing factors. We report two cases of true exfoliation that were confirmed after cataract surgery. Neither patient exhibited true exfoliation before cataract surgery. In addition, neither patient had a history of occupation with exposure to excessive heat, inflammation or trauma. We observed the anterior lens capsules of these two patients with slit-lamp microscopy before and after cataract surgery. True exfoliation disappeared by adhering to the anterior capsule in both cases, and there were no complications during the observation period.

Anterior chamber irrigation with an ozonated solution as prophylaxis against infectious endophthalmitis.

PURPOSE: To assess the validity of anterior chamber irrigation with an ozonated solution as prophylaxis against endophthalmitis. SETTING: Department of Ophthalmology, Nippon Medical School, Tokyo, Japan. METHODS: Viability of human corneal endothelium in culture was assessed by the WST-8 assay, lactate dehydrogenase (LDH) release assay, and trypan blue exclusion assay after exposure to a 4 to 40 parts per million (ppm) solution for 10 to 60 seconds. The in vivo effect was observed 1 week after irrigation of a 4 ppm solution in the rabbit anterior chamber by trypan blue exclusion assay. Bactericidal efficacy of the anterior chamber irrigation with the 4 ppm solution was examined by bacterial colony count of the aqueous humor following methicillin-resistant Staphylococcus aureus (MRSA) contaminated intraocular lens implantation in the porcine eye. RESULTS: The WST-8 assay revealed no significant reduction of viability with 10-second exposure to a 4 ppm solution. Lactate dehydrogenase release and trypan blue exclusion assays similarly demonstrated little damage after 60-second exposure to a 4 ppm solution. In the rabbit cornea 1 week after treatment, damage caused by 30-second exposure to a 4 ppm solution was not significant. The MRSA colony count documented almost complete bactericidal action with 5-second exposure to the 4 ppm solution when no ophthalmic viscosurgical device existed in the anterior chamber. CONCLUSION: Limited damage to the corneal endothelium after 10-second exposure and potent bactericidal action with 5-second exposure suggests the validity of anterior chamber irrigation with a 4 ppm ozonated solution as prophylaxis against endophthalmitis.

The effect of up- and downregulation of MnSOD enzyme on oxidative stress in human lens epithelial cells.

PURPOSE: Gene knockouts serve as useful experimental models to investigate the role of antioxidant enzymes in protection against oxidative stress in the lens. In the absence of gene knockout animals for Mn-containing superoxide dismutase (MnSOD), the effect of this enzyme on oxidative stress was investigated in a human lens epithelial cell line (SRA 01/04) in which the enzyme was up- or downregulated by transfection with sense and antisense expression vectors for MnSOD. METHODS: Human lens epithelial cells (SRA 01/04) were transfected with plasmids containing sense and antisense human cDNA for MnSOD. The enzyme levels were determined by Western blot analysis and immunocytochemistry. The protective effect of the enzyme against the cytotoxicity of H(2)O(2) was evaluated by cell viability, DNA strand breaks, and morphologic changes observed by transmission electron microscopy. In addition, the protective effect of this enzyme against photochemically induced stress and UVB irradiation in cells was assessed by measuring the damage of cellular DNA. RESULTS: The MnSOD level in upregulated cells was three times higher than in downregulated cells, and the enzyme surrounded the nucleus. Cells with elevated enzyme levels were more resistant to the cytotoxic effect of H(2)O(2) with greater cell viability. MnSOD-deficient cells showed dramatic mitochondrial damage when exposed to 50 micro M H(2)O(2) for 1 hour. Similarly, oxidative challenge by H(2)O(2), photochemically generated reactive oxygen species, or UVB irradiation produced greater DNA strand breaks in MnSOD-deficient cells than in those in which the enzyme was upregulated. CONCLUSIONS: These findings demonstrate the protective effect of MnSOD in antioxidant defense of cultured lens epithelial cells. This approach to modulating the enzyme level in cultured cells provides a new experimental model for study of the role of antioxidant enzymes in the lens.

Involvement of macrophage chemotactic protein-1 and interleukin-1beta during inflammatory but not basic fibroblast growth factor-dependent neovascularization in the mouse cornea.

Corneal neovascularization develops in several pathologic conditions, but its underlying mechanisms remain elusive. We used a mouse inflammatory corneal model (corneas cauterized with silver nitrate) and assessed the role of monocyte/macrophage-attracting factors, macrophage chemotactic protein-1 (MCP-1), and a proinflammatory cytokine, IL-1beta, on macrophage recruitment and neovascularization. Both MCP-1, IL-1beta protein, and mRNA levels increased markedly 12 hours after the chemical cauterization. In situ hybridization showed that MCP-1 was located in corneal epithelial cells, and IL-1beta was located in corneal epithelial cells and infiltrating inflammatory cells. In addition, double staining of corneas with antibodies specific for monocytes/macrophages and IL-1beta revealed that IL-1beta was found in infiltrating monocytes/macrophages at Day 2 after cauterization. Both IL-1beta and MCP-1 induced neovascularization in a rat cornea model, and the cauterization-induced corneal neovascularization was partially inhibited by subconjunctival injection of anti-IL-1beta or anti-MCP-1. Coadministration of two antibodies inhibited corneal neovascularization slightly more than that by the administration of each. In contrast, administration of the anti-MCP-1 or anti-IL-1beta showed minimal inhibition of basic fibroblast growth factor-driven corneal neovascularization by mouse cornea assay. Cauterized corneas treated with anti-MCP-1 antibody had significantly fewer monocytes/macrophages than control. These results indicate the existence of distinct monocyte/macrophage-involved angiogenic pathways in mouse cornea, in which MCP-1 released from corneal epithelial cells attracts monocytes/macrophages into the cornea, where they release IL-1beta leading to inflammatory neovascularization. In addition, the IL-1beta and MCP-1 released from the corneal epithelial cells may directly induce corneal neovascularization.

[Epidemiological study of outbreaks and sporadic cases due to Vibrio parahaemolyticus--serotype O3:K6 in Aichi Prefecture, Japan, during 1988 and 2001].

Percentage of the outbreaks by O3:K6 Vibrio parahaemolyticus (V. p) in Aichi Prefecture Japan increased from 3% (3/86) for 1988-95 to 75% (33/44) for 1996-2001. The percentage of the sporadic diarrhea cases caused by O3:K6 V. p in a general hospital in Aichi Prefecture also increased from 0% (0/253) to 61% (135/221) during the same periods. Thermostable direct hemolysin (TDH)-positive O3:K6 were isolated from 95% (19/20) of the outbreak incidents and 100% (135/135) of the sporadic cases. Only one TRH (TDH-related hemolysin)-positive O3:K6 was isolated from one outbreak incident. Percentage of the outbreaks by O3:K6 V. p associated with the consumption of boiled shellfishes increased from 5% (6/117) for 1988-95 to 25% (15/59) for 1996-2001, in particular, boiled crabs and squillas associated outbreaks increased from 2% (2/117) to 17% (10/59) and from 2% (2/117) to 10% (6/59), respectively. From 1,548 raw sea foods sampled in the Nagoya Central Wholesale Market in Aichi Prefecture in 1995-99, one TDH-positive O3:K6 was isolated from one live squilla (1/30). Increase in the percentage of outbreaks associated with TDH-positive O3:K6 V. p after 1996 in Aichi Prefecture was revealed to correlate with the increase in the outbreaks associated with consumption of boiled sea foods, especially boiled crabs as well as squillas. Accordingly, it becomes clear that sanitary handling of these boiled foods is important to prevent outbreaks and sporadic cases of diarrhea caused by O3:K6 V. p infection.