Comparison between a traditional single still image and a multiframe video image along the z‐axis of the same microscopic field of interest in cytology: Which does contribute to telecytology?

The limitation of cytologic still images is one of the reasons why telecytology has not met with widespread acceptance by the cytology community. Cytologic still image only displays a single depth of field, and this is a particularly acute problem in cytology where the specimen is often much thicker than a single microscopic depth of focus. In this article, we examine the validity of a “z‐axis” video of a microscopic field of interest. After observing videos of fields of interest from 10 cases, five cytotechnologists reached suitable cytologic findings and diagnosed the fields correctly in great majority of cases. Five other cytotechnologists, who looked only at a single representative still image, could not always make a correct diagnosis. The difference between two observer groups was statistically significant by Wilcoxon's matched pairs signed‐rank test. The results indicate that “z‐axis” video of microscopic field of interest provides a similar experience to “focusing through” observation of the specimen under a microscope and may improve an accuracy of primary telecytodiagnosis. And we expect that video image telecytology will strongly influence cytology, especially in education and training. Diagn. Cytopathol. 2009. © 2009 Wiley‐Liss, Inc.     

In vitro and in vivo Effects of Cisplatin and Etoposide in Combination on Small Cell Lung Cancer Cell Lines

The effects of cisplatin (CDDP) and etoposide (ETP) in combination were evaluated in vitro and in vivo using small cell lung cancer cell lines. The combination effects in vitro were investigated using isohologram analysis. Used together, CDDF and ETP showed a synergistic effect against cell growth on only 1 cell line (SBC-3), additive effects on 6 (SBC-2, SBC-5, Lul30, Lul34AH, Lul35T and H69) and an antagonistic effect on 1 (SBC-1). In the in vivo experiment, nude mice were inoculated with SBC-1, SBC-3 and SBC-5 cells. Two or 5 mgAg CDDP and 10 or 30 mgAg ETP were administered intraperitoneally alone and simultaneously in combination to nude mice. The in vivo effects of the combination were determined by comparing the observed growth ratio in mice treated with the combination with the expected value of this ratio calculated based on the assumption that the effects of the drugs were simply additive. According to this definition, synergistic effects were observed against all 3 tumors. Thus, the in vivo and in vitro effects differed. The toxicity of the combination therapy, which was analyzed by estimating the body weight change of mice, was no higher than that of CDDP or ETP alone. These results suggest that the excellent clinical effects of CDDP and ETP combination therapy may he attributable not to drug interaction at the cellular level hut to the feasibility of combined use of them at full doses without overlapping side effects.     

Synergistic Antimetastatic Effects of Lentinan and Interleukin 2 with Pre- and Post-operative Treatments

The antimetastatic activity of a combination of lentinan and interleukin 2 (IL-2) was evaluated against spontaneously metastatic 3-methylcholanthrene-induced DBA/2.MC.CS.T fibrosarcoma. Although pre-operative treatment with either IL-2 or lentinan alone exerted little effect on the reduction of lung metastasis colony numbers (7.1% or 28.4% reduction, respectively), the combination exhibited a synergistic effect (85% reduction). Furthermore, 3 of 13 mice given the pre-operative combination treatment achieved complete cure, while no mice given saline did. Although the post-operative combination treatment also reduced the colony number (71% reduction), it caused little prolongation of survival and no mouse achieved complete cure. Synergistic effects were observed between pre- and post-operative treatments with lentinan and IL-2: 8 of 12 mice were completely cured. The anti-metastatic activity was abolished in mice treated simultaneously with antibodies to CD4 and CDS antigens, whereas either CD4, CDS, or NK1.1 antibody alone was ineffective. Analysis of the cellular mechanism involved in the antimetastatic activity revealed the involvement of a tumor-associated antigen-specific delayed-type hypersensitivity response. These data suggest that the life-prolonging effect of the combination of lentinan and IL-2 is mediated by antigen-specific T cells and that the combination of pre- and post-operative therapy with lentinan and IL-2 may be effective to prevent cancer recurrence and metastasis after surgical resection.     

Inhibitory effect of bucillamine on laser-induced choroidal neovascularization in rats

PURPOSE: We investigated the inhibitory effects of bucillamine on formation of laser-induced choroidal neovascularization (CNV) in a rat model. METHODS: Bucillamine administration (approximately 150 mg/kg/day) was started 1 week before photocoagulation and continued to the end of the study. Control groups received drinking water. Two weeks after photocoagulation, choroidal neovascularization development was evaluated using simultaneous fluorescein and indocyanine green angiography, and the maximal thickness of the lesions was measured histologically. RESULTS: The incidence of CNV formation was 99.5 +/- 0.2% [mean +/- standard deviation (SD)] in control rats and 64.3 +/- 15.1% with bucillamine (P < 0.01). Histological study showed that the thickness of the CNV lesions was 23.4 +/- 6.5 microm (mean +/- SD) in the bucillamine-treated rats, which was significantly decreased compared to that in controls (60.8 +/- 9.2 microm) (P < 0.01). CONCLUSIONS: Our results suggest that bucillamine may inhibit the development of laser-induced CNV in rats.     

The effect of nitric oxide on retinal blood flow during hypoxia in cats

PURPOSE: To investigate how nitric oxide (NO) contributes to the regulation of retinal circulation during rest and hypoxia in cats. METHODS: N(G)-nitro-L-arginine-methyl ester (L-NAME; n = 7), an NO synthase inhibitor; N(G)-nitro-D-arginine methyl ester (D-NAME; n = 6), the inactive isomer; or phosphate-buffered saline (PBS; n = 7) was injected intravitreously. Hypoxia was induced in the cats by the administration of 10% oxygen. Vessel diameter and blood velocity were measured simultaneously in the anesthetized cats with a laser Doppler velocimetry system, and the retinal blood flow (RBF) and wall shear rate (WSR) were calculated as an index of shear stress on the retinal vessel wall. RESULTS: After intravitreous injection of L-NAME, the vessel diameter (-8.1% +/- 2.0%, P < 0.01), velocity (-17.0% +/- 3.7%, P < 0.01), and RBF (-29.4% +/- 4.6%, P < 0.01) significantly decreased compared with the preinjection level. In the PBS group, maximum increases above the prehypoxia diameter (13.7% +/- 3.5%, P < 0.01), velocity (39.5% +/- 8.6%, P < 0.01), RBF (73.9% +/- 10.6%, P < 0.01), and WSR (28.3% +/- 7.1%, P < 0.01) were observed during hypoxia. In the L-NAME group, those changes substantially decreased in response to hypoxia. D-NAME was inactive with regard to RBF during rest and hypoxia. CONCLUSIONS: These results suggest that NO may be continuously produced in the retina during rest and contributes to increased RBF during hypoxia. In addition, the increased WSR during hypoxia indicates that NO plays an important role in retinal hypoxic hyperemia through a flow-induced mechanism.     

Nobiletin as a tyrosinase inhibitor from the peel of Citrus fruit

A tyrosinase inhibitor was isolated from the peel of Citrus fruit by activity-guided fractionation, and identified as 3',4',5,6,7,8-hexamethoxyflavone (nobiletin) by comparison with reported spectral data. Nobiletin (IC50 of; 46.2 microM) exhibited more potency than Kojic acid (IC50; 77.4 microM) used as a positive control, and it was found to be potentially an effective inhibitor of the production of melanin.     

In vitro antifungal activity of naphthoquinone derivatives

In vitro antifungal activities of naphtoquinone-derivatives, which are constituents of Shikon, roots of Lithospermum erythrorhizon, were investigated against several fungal pathogens. When the biological activity of these compounds was tested against fungi, a wide range of sensitivity was recorded. Shikonin was found to have a stronger than fluconazole against yeast-like fungi: four-fold against Candida krusei (minimal inhibitory concentration (MIC); 4 microg/ml) and two-fold (MIC; 4 microg/ml) against Saccharomyces cerevisiae, though it showed the same potency as fluconazole against C. glabrata. Deoxyshikonin also exhibited four-fold stronger activity against C. krusei (MIC; 4 microg/mi) and three-fold (MIC; 2 microg/ml) stronger against S. cerevisiae. Acetylshikonin and beta-hydroxyisovaleryl shikonin showed lower activities against all fungal pathogens except for C. krusei compared with the standard. Against the filamentous fungus, Trichosporon cutaneum, all naphthoquinones were found to have a range of activity with lower potency than standard. This result provides a rational basis for the clinical use of shikon and shows the possibility of its use in medicinal treatment as an anti-inflammatory agent with antifungal activity.     

Successful treatment of multicentric reticulohistiocytosis with alendronate: evidence for a direct effect of bisphosphonate on histiocytes

We describe the case of a 44-year-old Japanese woman with severe nodular erythematous skin lesions and arthritis mutilans who was admitted for further treatment of multicentric reticulohistiocytosis. Skin and synovial biopsies showed heavy infiltration with tartrate-resistant acid phosphatase-positive histiocytes and multinucleated giant cells. Immunohistochemical analysis showed that some of the mononuclear cells in the skin were positive for RANKL. After 1 month of Alendronate, an aminobisphosphonate, given at a dosage of 10 mg once a week intravenously for the first 6 weeks and then once a month thereafter, the arthritis and skin nodules improved, and the remission has continued for more than 2 years. The findings in this patient suggest that osteoclast-like multinucleated giant cells differentiate locally in the skin from infiltrating histiocytes with the help of RANKL-positive stromal cells and that alendronate acts directly on cells of monocyte/macrophage lineage in humans. Thus, alendronate should be added to the list of drugs for the treatment of multicentric reticulohistiocytosis.