A natural infection by the red sea bream iridovirus-type Megalocytivirus in the golden mandarin fish Siniperca scherzeri.

An outbreak of a Megalocytivirus infection was found in the golden mandarin fish Siniperca scherzeri during September and October 2016, in Korea. Phylogeny and genetic diversity based on the major capsid protein (MCP) and adenosine triphosphatase (ATPase) genes showed a new strain. Designated as GMIV, this strain derived from the golden mandarin fish was suggested to belong to the red sea bream iridovirus (RSIV)-subgroup I. Additionally, this train clustered with the ehime-1 strain from red sea bream Pagrus major in Japan and was distinguished from circulating isolates (RSIV-type subgroup II and turbot reddish body iridovirus [TRBIV] type) in Korea. The infection level, evaluated by qPCR, ranged from 8.18 × 102 to 7.95 × 106  copies/mg of tissue individually, suggesting that the infected fish were in the disease-transmitting stage. The diseased fish showed degenerative changes associated with cytomegaly in the spleen as general sign of Megalocytivirus infection. The results confirm that the RSIV-type Megalocytivirus might have crossed the environmental and species barriers to cause widespread infection in freshwater fish.

Topical calcitriol restores the impairment of epidermal permeability and antimicrobial barriers induced by corticosteroids.

BACKGROUND: The active form of vitamin D(3) , calcitriol, is widely used for the treatment of psoriasis, with or without topical corticosteroids. Topical corticosteroids are known to disrupt permeability and antimicrobial barriers, even with short-term use. Yet, the effect of topical calcitriol on epidermal permeability and antimicrobial barriers disrupted by topical corticosteroids has not been determined. OBJECTIVES: To examine the effect of calcitriol on epidermal permeability and antimicrobial barrier function that has been impaired by corticosteroids, as well as to elucidate the mechanism of improvement. MATERIAL AND METHODS: Topical calcitriol or the control vehicle was applied to each flank of hairless mice 20 min after treatment with topical clobetasol propionate and repeated every 12 h for 3·5 days. Barrier function assessment, Nile red staining, electron microscopy, immunohistochemistry, Western blotting, and real-time reverse transcriptase-polymerase chain reaction studies were performed 24 h after the last application. RESULTS: Epidermis co-treated with topical calcitriol showed an improvement of stratum corneum integrity and barrier recovery, more intense fluorescence staining with Nile red, and an increase in lamellar body (LB) maturation and density, as well as upregulation of major epidermal lipid synthesis-related enzymes (3-hydroxy-3-methylglutaryl-CoA, serine-palmitoyl transferase and fatty acid synthase), mouse beta-defensin 3, cathelin-related antimicrobial peptide and vitamin D receptor. CONCLUSIONS: We found that topical calcitriol restored both the epidermal permeability and antimicrobial barrier that had been impaired by corticosteroids. This restoration was mediated by both an activation of the cutaneous vitamin D pathway and an increase of epidermal lipids and antimicrobial peptides, promoted by the formation of the LB and the activity of epidermal lipid synthesis-related enzymes.

Novel strategy for a bispecific antibody: induction of dual target internalization and degradation.

Activation of the extensive cross-talk among the receptor tyrosine kinases (RTKs), particularly ErbB family-Met cross-talk, has emerged as a likely source of drug resistance. Notwithstanding brilliant successes were attained while using small-molecule inhibitors or antibody therapeutics against specific RTKs in multiple cancers over recent decades, a high recurrence rate remains unsolved in patients treated with these targeted inhibitors. It is well aligned with multifaceted properties of cancer and cross-talk and convergence of signaling pathways of RTKs. Thereby many therapeutic interventions have been actively developed to overcome inherent or acquired resistance. To date, no bispecific antibody (BsAb) showed complete depletion of dual RTKs from the plasma membrane and efficient dual degradation. In this manuscript, we report the first findings of a target-specific dual internalization and degradation of membrane RTKs induced by designed BsAbs based on the internalizing monoclonal antibodies and the therapeutic values of these BsAbs. Leveraging the anti-Met mAb able to internalize and degrade by a unique mechanism, we generated the BsAbs for Met/epidermal growth factor receptor (EGFR) and Met/HER2 to induce an efficient EGFR or HER2 internalization and degradation in the presence of Met that is frequently overexpressed in the invasive tumors and involved in the resistance against EGFR- or HER2-targeted therapies. We found that Met/EGFR BsAb ME22S induces dissociation of the Met-EGFR complex from Hsp90, followed by significant degradation of Met and EGFR. By employing patient-derived tumor models we demonstrate therapeutic potential of the BsAb-mediated dual degradation in various cancers.

An 86 kDa diapause protein 1-like protein is a component of early-staged encapsulation-relating proteins in coleopteran insect, Tenebrio molitor larvae.

Recently, we reported two novel early-staged encapsulation-relating proteins (56 kDa and 48 kDa ERPs) isolated from the hemolymph of coleopteran insect, Tenebrio molitor larvae [Cho et al. (1999) Eur. J. Biochem. (in press)]. Here, a cDNA clone for another early-staged encapsulation-relating protein (86 kDa) was isolated. We found that the 86 kDa protein shows high homology with insect diapause protein 1. The 86 kDa protein was localized in the fat body and hemolymph, but not hemocyte lysate. A significant level of 86 kDa protein was detected in pre-pupae stage, but it decreased rapidly at late larvae and pupae, and no protein was found in embryo, early larvae and adult stages. This diapause protein 1-like protein is likely to be a component of early-staged encapsulation-relating proteins in the insect cellular defense reaction.

The pro-phenoloxidase of coleopteran insect, Tenebrio molitor, larvae was activated during cell clump/cell adhesion of insect cellular defense reactions.

To characterize the proteins involved in cell clump/cell adhesion of insect cellular defense reactions, we induced the cell clump/cell adhesion reaction in vitro with the hemolymph of larvae of the coleopteran insect, Tenebrio molitor. The 72 kDa protein was specifically enriched in the residues of cell clump/cell adhesion and was purified to homogeneity. A cDNA clone for the 72 kDa protein was isolated. We found that the 72 kDa protein was an activated phenoloxidase from Tenebrio pro-phenoloxidase. We suggest that activated phenoloxidase is involved in the cell clump/cell adhesion reaction as well as in the synthesis of melanin.

Calreticulin enriched as an early-stage encapsulation protein in wax moth Galleria mellonella larvae.

To investigate the molecular mechanism of the early-stage encapsulation reaction in insects, we purified a 47kDa protein from injected beads into Galleria mellonella larvae. When a cDNA clone was isolated, the 47kDa protein showed high homology with Drosophila and human calreticulin. Western blotting analysis showed that the 47kDa protein was present in the hemocytes, but not in the plasma. When the early-stage encapsulated beads were coated with 47kDa protein antibody and reinjected into G. mellonella larvae, any further encapsulation reaction was inhibited. These results suggest that calreticulin is involved in non-self recognition in invertebrate cellular defense reactions.

Chemically induced rat mammary tumor treated with tamoxifen showed decreased expression of cyclin D1, cyclin E, and p21(Cip1).

We studied the effects of tamoxifen (TAM) on the growth of 7,12-dimethylbenz(a)anthracene (DMBA)-induced rat mammary tumor and the expression of cyclin D1, cyclin E, p21(Cip1), and estrogen receptors (ER) by performing immunohistochemistry and Western blot analysis. When tumor size reached between 10 and 15mm in the largest dimension, the rats were divided into a DMBA-control group and a DMBA-TAM group. The administration of TAM markedly decreased the tumor development and showed decreased expression of bromodeoxyuridine, cyclin D1, cyclin E, and p21(Cip1) when compared with those of the DMBA-control group; however, a few tumors showed progressive growth in spite of TAM treatment. These tumors had decreased expression of ER. This study suggests that TAM suppresses tumor development through the down-expression of cyclin D1 and cyclin E.

Purification and cloning of glyoxalase II from rat liver.

Glyoxalase (GLO) II, which is a component of GLO system and catalyze the conversion of S-lactoyl-glutathione to D-lactate, was purified 1488 fold from rat liver by two steps of Affigel blue and carbobenzoxyglutathione-Sepharose 4B affinity chromatography. The molecular weight of the enzyme was estimated to be 29 kDa which is similar to those from other species. The sequence of N-terminal 9 amino acid residues was determined to be MGIRLLPAT. This was then used to synthesize degenerative primers. cDNA clone was isolated by first synthesizing cDNA from RNA and then PCR amplification. The sequence of cDNA clone was determined by serial sequencing analysis.

Regulation of leptin gene expression by insulin and growth hormone in mouse adipocytes.

The role of leptin in the control of obesity, insulin resistance and type II diabetes has been reported, however, the regulatory mechanism of leptin in animals affected by hormones is not clearly understood. In this study, the effects of insulin, epinephrine, growth hormone or dexamethasone on the expression of leptin was examined in mouse primary adipocytes. The leptin expression was also studied in the adipose tissue of the mouse treated with insulin or growth hormone (0.3 or 0.6 units/animal). Insulin (100 nM) or dexamethasone (100 nM) stimulated leptin mRNA transcription while epinephrine (100 nM) alleviated its transcription in mouse primary adipocytes. The level of leptin protein in cultured media of adipocytes treated with insulin or dexamethasone was higher than that of the control group but growth hormone or epinephrine treatment had no effect on them. Insulin administration (0.6 units/mouse) enhanced leptin mRNA as well as leptin protein in mouse adipose tissue but growth hormone administration (0.3 or 0.6 units/mouse) had no effect on them. Leptin protein level in sera of mice injected with insulin or growth hormone was not significantly different from that of control group. These results indicate that both insulin and dexamethasone stimulate leptin gene expression and secretion of its product, whereas, growth hormone has no effect on the expression of leptin gene in mouse adipocytes.

The effect of UVA irradiation on the epidermal melanocyte after 8-MOP and TMP administration.

Morphological and numerical changes in the epidermal melanocytes of black C57BL mice after phototoxic drug administration followed by ultraviolet A irradiation were studied to compare the effects of photochemotherapy on the epidermal melanocytes using 8-methoxypsoralen and trimethylpsoralen. One hour after intraperitoneal injection of the phototoxic drugs, 1.5 mg/kg in the small dose group and 6.0 mg/kg in the large dose group, the mice were exposed to UVA irradiation. This procedure was performed twice a week for 8 weeks at the small dose group and for 5 weeks in the large dose group. Skin biopsies were taken before irradiation in both groups, and follow up biopsies were done at each week. The number and size of the melanocytes were observed in a split-DOPA preparation. In the drug treated groups, there was an increase in the size of the perikaryon, and the number, length, width, and arborization of dendrites. Such changes were more clearly seen in the group treated with trimethylpsoralen compared with the 8-methoxypsoralen treated group. Therefore, trimethylpsoralen is more effective than 8-methoxypsoralen in the increase of the perikaryon size, and the number, length, width, and arborization of dendrites of melanocytes in the intraperitoneal injection.

The fate of small-diameter homografts in the pulmonary position.

BACKGROUND: Limited durability is expected for small homograft valves that are used to correct congenital cardiac disease. METHODS: All 76 homograft valves with an internal annulus diameter ranging from 8 to 13 mm that were implanted from 1987 through 2000 in the pulmonary position were retrospectively analyzed. In each case, homograft size was normalized to the patient's body surface area: z-value. For 93% (14 of 15) of the 8 to 9 mm grafts, z was less than 2. For 56% (5 of 9) of the 10 mm grafts and 98% (51 of 52) of the 11 to 13 mm allografts, z was greater than 2. Survival and freedom from complications were estimated by the Kaplan-Meier method. Homograft failure was defined as homograft replacement or late death; significant dysfunction, as homograft obstruction with an echo-Doppler gradient greater than 50 mm Hg or grade III or IV valvular insufficiency. The log-rank test was used to compare outcomes. RESULTS: Seven patients died early after operation; three, late. Survival was 86.5% +/- 3.8% at 1 year and remained stable during the succeeding years. Freedom from failure for all homografts was 90.6% +/- 3.7%, 71.8% +/- 6.9%, and 61.8% +/- 9.0% at 1, 5, and 10 years, respectively. Corresponding freedom from significant dysfunction was 87.6% +/- 4.1%, 51.2% +/- 7.4%, and 10.1% +/- 8.3%. The smaller homografts (z less than 2) failed and deteriorated faster (p < 0.0001): only 32.1% +/- 13.0% were still functioning at 24 months. The larger grafts (z at least 2) retained function for the first 4 years, and 73.7% +/- 10.4% had not yet failed at 10 years. CONCLUSIONS: Smaller (z less than 2) homografts (the great majority of 8 to 9 mm grafts) have to be replaced early, usually within 2 years of implantation. Larger (z at least 2) grafts (nearly all 11 to 13 mm grafts) show remarkable durability and are suitable valved conduits for establishing right ventricle to pulmonary artery continuity in neonates and young infants.

Comparison of the efficacy and safety of different formulations of omeprazole-based triple therapies in the treatment of Helicobacter pylori-positive peptic ulcer.

Little is known about the efficacy and safety of different formulations of omeprazole-based triple therapy regimens for the treatment of Helicobacter pylori-positive peptic ulcer. We compared the efficacy and safety of two formulations of omeprazole used in triple therapies in patients with H. pylori-positive active peptic ulcer. Seventy-four patients with endoscopically proven H. pylori-positive active peptic ulcer were randomized to two groups, each with 37 patients, to receive either OAC-I (6 weeks of "A" formulation of omeprazole [20 mg twice daily] plus 2 weeks of amoxicillin [1.0 g twice daily] and clarithromycin [500 mg twice daily] or OAC-II (6 weeks of "B" formulation of omeprazole [20 mg twice daily] plus 2 weeks of the same antibiotics. The H. pylori and ulcer healing status were assessed at the baseline and at the 6-week endpoint of therapy. Gastrointestinal symptoms, documentation of adverse events, and standard laboratory examinations were assessed at each visit. Eradication of H. pylori (intention to treat [n = 74]/per protocol [n = 66]) and healing of the ulcer were successful in 83.8%/96.9% and 93.8%, respectively, of the OAC-I group patients, and in 91.9%/100% and 97.1%, respectively, of the OAC-II group patients (P = 0.477; P = 0.608). The OAC-I group experienced rapid resolution of symptoms, but no significant differences were found between the two groups for number of days taken for resolution of gastrointestinal symptoms, adverse events, and laboratory findings. The two different formulations of omeprazole used in triple therapy regimens produced similar efficacy and safety results after 6 weeks of treatment in patients with H. pylori-positive active peptic ulcer.

T-lymphocyte subsets in patients with AJCC stage III gastric cancer during postoperative adjuvant chemotherapy. American Joint Committee on Cancer.

BACKGROUND AND AIMS: Advanced neoplastic diseases alter the immune response in cancer patients. The aim of this study was to evaluate the changes of T-lymphocyte subsets during postoperative adjuvant chemotherapy, and the relationship between T-lymphocyte subsets and tumor recurrence in AJCC stage III gastric cancers. MATERIAL AND METHODS: Analysis of T-lymphocyte subsets was performed in 39 patients with stage III gastric adenocarcinoma who had undergone a curative gastric resection and postoperative chemotherapy. CirculatingT-lymphocyte subsets were measured on venous blood by using flow cytometry and monoclonal antibodies on preoperative day 1, and postoperative months 1, 3, and 6. RESULTS: The 5-year disease-free survival rates of patients with stage 3a and 3b gastric cancer were 57.1% and 33.3%, respectively (p = 0.06). Values of CD3+ and CD4+ T-cells, and CD4+/CD8+ ratios were consistently lower in the recurrence group throughout the observation period. CD4+ T-cell counts were significantly lower in the recurrence group on preoperative day 1, and postoperative months 1 and 6. However, most values of the T-lymphocyte subsets showed no statistically significant difference when comparing the stage 3a and 3b disease patient groups. CONCLUSIONS: The results of this study suggest that immunosuppression associated with CD3+ and CD4+ T-cell depression is a risk factor for postoperative recurrence in patients with stage III gastric cancer.

Expression of eosinophil chemotactic factors in stomach cancer.

We have occasionally experienced eosinophilic abscess of the liver in patients with gastric carcinoma, suggesting that some eosinophil mobilizing (chemotactic and proliferative) factors might be produced by carcinoma cells. The aim of this study was to determine whether or not gastric carcinoma expresses the well-known eosinophil chemotactic factors (ECFs) and whether or not the expression is related to the histologic subtypes. Seventeen consecutive surgically removed tumor-bearing stomachs were collected: 7 signet ring cell type, 7 poorly differentiated tubular adenocarcinoma, and 3 moderately differentiated tubular adenocarcinoma. Hematoxylin-eosin stained sections were re-evaluated for eosinophil and mast cell infiltration. The expression of IL-2, IL-5 and granulocyte-macrophage colony stimulating factor (GM-CSF) were examined by immunocytochemical stain. There was no available frozen tissue for IL-2 and IL-5 in one case. Gastric carcinoma expressed IL-2 in all 16 cases, IL-5 in 12 of 16 cases and GM-CSF in 10 of 17 cases. Of particular interest, 7 of 10 GM-CSF-expressing carcinomas were signet ring cell type. Even in the remaining 3 cases, most GM-CSF-positive cells were signet ring cells scattered within tubular adenocarcinoma. No correlation of ECF expression between either eosinophil/mast cell infiltration or peripheral blood eosinophilia was identified. In conclusion, most gastric carcinomas express the well-known ECFs and the expression of GM-CSF is specific for signet ring carcinoma cells.

p53 protein overexpression in astrocytic neoplasms.

Abnormalities of the p53 gene are the most common molecular change in human cancer. In the central nervous system, mutant p53 gene is frequently identified in the tumors with astrocytic differentiation. To investigate the relation between histologic subtypes and p53 protein overexpression, we examined 81 cases of astrocytic neoplasms (24 benign astrocytoma, 28 anaplastic astrocytoma and 29 glioblastoma multiforme) using the standard immunohistochemical method. All were formalin-fixed and paraffin-embedded tissue. The p53 immunoreactivity was found in 4/24 benign astrocytoma, 18/28 anaplastic astrocytoma, 22/29 glioblastoma multiforme. The degree of immunoreactivity closely correlated with histologic subtypes (p < 0.001). Overall p53 protein expression was most frequently detected in glioblastoma multiforme, but strong immunoreactivity (3+) was more frequently found in the anaplastic astrocytoma than in glioblastoma multiforme. Although the frequency of p53 protein expression is low, 4 benign astrocytoma showed distinct nuclear staining. In conclusion the malignant progression of astrocytic neoplasms may be associated with increasing expression of p53 protein.

Malignant granular cell tumor at the retrotracheal space.

We report a case of an extremely rare neoplasm, malignant granular cell tumor (MGCT). The patient was a 21-year-old woman, who was 5 months pregnant. The tumor occurred in the retrotracheal space, extending from the level of the larynx to the thoracic inlet. In addition, there were multiple, variable-sized tumor nodules within both lung fields on chest CT scan. Histologically, tissue biopsied from the periphery of the tumor consisted of solid sheets of large ovoid cells with ample, eosinophilic cytoplasm, eccentric nuclei, and prominent nucleoli. Each cell showed slight atypism of the nuclei. There was a focal necrosis at the periphery of the lesion. These cells stained strongly for S-100 protein, neuron-specific enolase (NSE) and CD68. On electron microscopy, the tumor cells contained autophagic vacuoles. The patient refused further treatment and died 7 months later. The exact cause of death was not known. Until now, the diagnosis of MGCTs has been made only when metastasis and an aggressive clinical course are identified, although some observers advocate that some histologic features such as nuclear pleomorphism, necrosis, and the presence of any mitotic activity are indicative of malignancy. These histologic findings are not easily detectable in every case of MGCT, as in our case. So the diagnosis of a MGCT should be considered in cases with aggressive clinical findings and some histologic features, such as necrosis, nuclear atypism, and mitotic activities, which could suggest the malignant behavior of this neoplasm.

Study of the skin concentrations after administration of the various phototoxic drugs.

The skin concentrations of 8-methoxypsoralen (8-MOP), 5-methoxypsoralen (5-MOP), and 4, 5', 8-trimethylpsoralen (TMP) were studied in the guinea pig following oral administration and bathing. The skin concentration of phototoxic drugs after oral administration peaked at 1.5 hours, and the concentration of 8-MOP was 3.5 times greater than that of 5-MOP. The skin concentration of TMP was not detected in our study (limit of sensitivity 5ng/ml). The skin concentrations of phototoxic drug after bathing decreased in the order of 5-MOP, TMP, and 8-MOP.

Endoscopic mucosal resection with a ligation device for early gastric cancer and precancerous lesions: comparison of its therapeutic efficacy with surgical resection.

Endoscopic mucosal resection with a ligation device (EMR-L) has become important in the curative treatment of precancerous lesions and early gastric cancers (EGCs), but little is known of the long-term efficacy and survival rates of EMR-L compared with surgical resection. We analyzed the therapeutic efficacy and safety of EMR-L in cases of EGC and precancerous lesions and compared the results of EMR-L with those of gastrectomy in patients with EGC over the same periods. EMR-L was performed on 20 EGCs and 54 precancerous lesions including tubular adenomas with or without severe dysplasias in 74 patients. Macroscopic type, size and location of the lesion were determined by endoscope, and the depth of invasion in EGCs was determined by endoscopic ultrasonography and confirmed by pathologic examination of the resected specimens. All the EGC cases were endoscopically followed up for at least 18 months (range, 18-66 months). Patients were selected that underwent subtotal gastrectomy and the survival rates were compared with those that underwent EMR-L. Complete resection was made in a single EMR-L treatment session in 61 cases (82.4%; 91.5%, were precancerous lesions and 65% were EGCs). After a repeat trial of EMR-L, the total rate of complete resection of precancerous lesions and EGCs was 92.6% and 85.0%, respectively. The survival rate of EGCs showed that complete resection by EMR-L resulted in 2 and 5 year survival rates of 100% and 95%, which are comparable to those of surgery (100% and 100%). This study suggests that EMR-L is a technically simple, minimally invasive and highly safe and effective treatment modality for selective EGCs, and offers an alternative to surgical treatment.

Treatment of vitiligo with oral 5-methoxypsoralen.

Thirty-six patients with vitiligo were treated with oral 5-methoxypsoralen (5-MOP) and subsequently exposed to UVA irradiation. The patients were treated once or twice weekly over a period of 2-10 months, taking 40-60 mg of 5-MOP 2 hours before exposure to UVA light. The amount of exposure to UVA light was slowly increased according to the patient's tolerance. Eleven (31%) patients showed remarkable repigmentation of the areas of vitiligo within six months. Overall, 78% of the patients showed effective repigmentation. Areas of vitiligo on the face and trunk were more responsive to treatment than those on the distal part of limbs. Adverse effects due to the drug included 2 patients with nausea and 1 patient with headache. It is suggested that treatment with systemic 5-MOP is effective, safe, and useful in selected cases of vitiligo.

Dosimetric advantages of generalised equivalent uniform dose-based optimisation on dose-volume objectives in intensity-modulated radiotherapy planning for bilateral breast cancer.

OBJECTIVE: We compared and evaluated the differences between two models for treating bilateral breast cancer (BBC): (i) dose-volume-based intensity-modulated radiation treatment (DV plan), and (ii) dose-volume-based intensity-modulated radiotherapy with generalised equivalent uniform dose-based optimisation (DV-gEUD plan). METHODS: The quality and performance of the DV plan and DV-gEUD plan using the Pinnacle(3) system (Philips, Fitchburg, WI) were evaluated and compared in 10 patients with stage T2-T4 BBC. The plans were delivered on a Varian 21EX linear accelerator (Varian Medical Systems, Milpitas, CA) equipped with a Millennium 120 leaf multileaf collimator (Varian Medical Systems). The parameters analysed included the conformity index, homogeneity index, tumour control probability of the planning target volume (PTV), the volumes V(20 Gy) and V(30 Gy) of the organs at risk (OAR, including the heart and lungs), mean dose and the normal tissue complication probability. RESULTS: Both plans met the requirements for the coverage of PTV with similar conformity and homogeneity indices. However, the DV-gEUD plan had the advantage of dose sparing for OAR: the mean doses of the heart and lungs, lung V(20) (Gy), and heart V(30) (Gy) in the DV-gEUD plan were lower than those in the DV plan (p<0.05). CONCLUSIONS: A better result can be obtained by starting with a DV-generated plan and then improving it by adding gEUD-based improvements to reduce the number of iterations and to improve the optimum dose distribution. Advances to knowledge The DV-gEUD plan provided superior dosimetric results for treating BBC in terms of PTV coverage and OAR sparing than the DV plan, without sacrificing the homogeneity of dose distribution in the PTV.