Exopolysaccharides from Lactobacillus delbrueckii ssp. bulgaricus OLL1073R-1 prevent influenza virus infection and attenuate secondary bacterial infection risk.

The present study assessed the inhibitory action of exopolysaccharides (EPS) produced by Lactobacillus delbrueckii ssp. bulgaricus OLL1073R-1 against influenza virus infection followed by secondary bacterial infection. We found that the presence of 200 or 400 µg ml-1 of EPS significantly protected against influenza virus infection in a dose-dependent manner when A549 cells were treated with EPS before infection but not after it. The expression of carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM-1), an adhesion molecule for bacteria adherence, on A549 cells was significantly enhanced during influenza virus infection compared with viral-non-infected A549 cells. However, this upregulated CEACAM-1 expression was significantly decreased by EPS treatment before viral infection in association with the reduction in the virus titre in A549 cells. In a bacterial adhesion assay using Staphylococcus aureus, the bacterial adherence to viral-infected A549 cells was significantly greater than that to viral-non-infected A549 cells, and the increased bacterial adherence induced by influenza virus infection tended to be decreased by EPS treatment before the infection. Our findings show that EPS treatment before viral infection can inhibit influenza virus infection and alleviate secondary bacterial infection through decreased CEACAM-1 expression.

Immunization with outer membrane protein A from Salmonella enterica serovar Enteritidis induces humoral immune response but no protection against homologous challenge in chickens.

Vaccination of poultry is one promising strategy to mitigate Salmonella infection in poultry and, in turn, humans as well. We evaluated the efficacy of outer membrane protein A (OmpA) as a novel vaccine candidate against Salmonella in poultry. Native OmpA purified from Salmonella enterica serovar Enteritidis was mixed with adjuvant and administered intramuscularly to 41-d-old chicks. The vaccinated birds showed no decrease in cecal excretion and tissue colonization compared with the unvaccinated birds after oral challenge with 10(9) cfu of the homologous strain at 28 d postimmunization. However, this vaccination induced an increased level of serum anti-OmpA IgG. Similar results were obtained in the replication experiments using a recombinant OmpA with single and double doses. For the development of more effective component vaccines for avian salmonellosis, the vaccine efficacy of outer membrane proteins other than OmpA and route of immunization other than parenteral administration should be evaluated with regard to protection and immune responses, including mucosal IgA.

TEL-Syk fusion constitutively activates PI3-K/Akt, MAPK and JAK2-independent STAT5 signal pathways.

We previously reported the fusion of the TEL gene to the Syk gene in myelodysplastic syndrome with t(9;12)(q22;p12). TEL-Syk fusion transformed interleukin-3 (IL-3)-dependent murine hematopoietic cell line BaF3 to growth factor independence. Here, we investigate the intracellular signal transduction of the stable transfectants. TEL-Syk fusion protein was associated with the p85 subunit of phosphatidyl inositol 3 kinase (PI3-K) followed by the activation of Akt in the absence of IL-3. Vav, phospholipase C-gamma2 and mitogen-activated protein kinase (MAPK) were also constitutively activated. TEL-Syk also activated the signal transducer and activator of transcription 5 (STAT5) in the absence of Janus kinase 2 activation. None of these kinases were phosphorylated in the BaF3 cells transfected with TELDeltaPNT-Syk in which the oligomerization domain of TEL was deleted. Inhibitor analysis showed that the MAPK pathway was important in TEL-Syk-mediated cell proliferation. The immunofluorescence technique revealed that the TEL-Syk fusion protein was located in the cytoplasm. These data suggest that TEL-Syk fusion protein in the cytoplasm leads to the constitutive activation of PI3-K/Akt, MAPK and STAT5 signal pathways, which are closely involved in IL-3-independent cell proliferation of BaF3 cells.

New procedure for purse-string suture in thoracoscopic esophagectomy with intrathoracic anastomosis.

BACKGROUND: In endoscopic surgery, one of the greatest problems is the difficulty with the reconstructive procedure. This problem frequently makes operating times longer. The authors have performed thoracoscopic esophagectomy and intrathoracic esophagogastric anastomosis for reconstruction using a circular stapler for the esophageal cancer. Although the circular stapler is a useful device for gastrointestinal anastomosis, it was difficult to place a purse-string suture and to fixate the anvil into the proximal esophagus endoscopically. METHODS: The authors devised a new procedure for the placement of the purse-string suture by using an Endo-Stitch device along with a new method to incise the esophageal wall and thereby facilitate fixation of the anvil. RESULTS: The authors attempted this procedure for five patients. The anastomoses were performed successfully. CONCLUSIONS: The new procedure can make endoscopic intrathoracic anastomosis feasible and safe. In addition, this procedure can be applied widely to other endoscopic reconstructions.

Audit of the use of psoralen photochemotherapy (PUVA) and narrowband UVB phototherapy in the treatment of psoriasis.

BACKGROUND: Psoralen photochemotherapy (PUVA), the combined use of psoralen and long wave ultraviolet (UVA) irradiation, was introduced around 1974 and its beneficial effects were rapidly confirmed worldwide. In an attempt to minimize its recognized long-term photocarcinogenic risk after some 150-200 exposures while also maintaining efficacy, however, the narrowband (311-312 nm) ultraviolet B (UVB) lamp (TL-01) was introduced in 1984, and has moved towards replacing PUVA except for severe or resistant disease. AIMS: To discover whether our use of these therapies complied with established British Photodermatology Group guidelines for PUVA and guidelines formulated within our unit for narrowband UVB. METHODS: The study was retrospective over 6 months from November 2001 to April 2002, all relevant information being obtained from the patients' hospital notes. RESULTS: Thirty-one patients received PUVA (18 oral, 11 bath and two uncertain because of missing notes) and 20 narrowband UVB during this period. CONCLUSIONS: Our PUVA and narrowband UVB phototherapy guidelines were shown to have been followed relatively closely with the following exceptions: one PUVA patient received a high cumulative exposure by mutual agreement because there was no other suitable therapy; a failure to measure minimal phototoxic doses (MPDs) in some PUVA patients; and slightly prolonged referral delays, but generally by patient choice.

Different mechanisms between premitotic apoptosis and postmitotic apoptosis in X-irradiated U937 cells.

PURPOSE: Apoptosis is currently being evaluated for its importance as a pathway of radiation-induced cell death. However, the difference in the mechanisms between premitotic and postmitotic apoptosis following X-irradiation remains not well understood. We show here that the human monoblastoid cell line U937 can be induced to undergo these two different types of apoptosis. METHODS AND MATERIALS: U937 cells were irradiated at a dose of 5 or 20 Gy, and the DNA fragmentation rate was measured by both flow cytometric analysis and gel electrophoresis. Activation of caspase-3 was detected by Western blot analysis and fluorogenic assay using acetyl-Asp-Glu-Val-Asp-7-amino-4-methyl-coumarin (Ac-DEVD-AMC). Detection of mitochondrial transmembrane potential (DeltaPsi) was performed by using Rho123. Chasing of S-phase fraction following X-irradiation was performed after labeling with 5-bromo-2'-deoxyuridine (BrdU). Thymidine was used for synchronization of the cells. Inhibition of caspase-3 activity was achieved by Acetyl-Asp-Glu-Val-Asp-aldehyde (Ac-DEVD-CHO). RESULTS: Time courses of the apoptotic rates, caspase activation, and DeltaPsi indicated that two different types of cell death were induced by the different X-ray doses. High-dose X-ray (20 Gy) induced a rapid and strong apoptosis, whereas low-dose X-ray (5 Gy) induced a slow and mild apoptosis. Cell-cycle analyses revealed that there was cell death before cell division in the former apoptosis but the cells must be dying after cell division in the latter apoptosis. By means of cell-cycle synchronization, the S-phase cells proved to be the most sensitive fraction to premitotic apoptosis, but an obvious difference in the susceptibility to cell death among the cell-cycle phases was not observed in postmitotic apoptosis. Ac-DEVD-CHO treatment effectively blocked caspase activity and premitotic apoptosis, but it failed to block postmitotic apoptosis. CONCLUSIONS: Irradiation of U937 cells at different X-ray doses induced two different types of apoptotic cell death, premitotic apoptosis and postmitotic apoptosis, which are characterized by the time course and cell-cycle specificity. Decision concerning these two types of apoptotic cell death may be made by the difference in the magnitude of cell damage following X-irradiation.

Measurement of direct photon emission in K+-->pi(+)pi(0)gamma decay

We have performed a measurement of the K+-->pi(+)pi(0)gamma decay and have observed 2x10(4) events. The best fit to the decay spectrum gives a branching ratio for direct photon emission of (4.7+/-0.8+/-0. 3)x10(-6) in the pi(+) kinetic energy region of 55 to 90 MeV and requires no component due to interference with inner bremsstrahlung.

Further search for the decay K+-->pi(+)nunu;

A search for additional evidence for the rare kaon decay K+-->pi(+)nunu; has been made with a new data set comparable in sensitivity to the previous exposure that produced a single event. No new events were found in the pion momentum region examined, 211pi(+)nunu;) = 1.5(+3.4)(-1.2)x10(-10).

Measurement of structure-dependent K+ --> &mgr;(+)nu(&mgr;)gamma decay

We report the first measurement of a structure-dependent component in the decay K+-->&mgr;(+)nu(&mgr;)gamma. Using the kinematic region where the muon kinetic energy is greater than 137 MeV and the photon energy is greater than 90 MeV, we find that the absolute value of the sum of the vector and axial-vector form factors is |F(V)+F(A)| = 0.165+/-0.007+/-0.011. This corresponds to a branching ratio of B(SD+) = (1.33+/-0.12+/-0.18)x10(-5). We also set the limit -0. 04

Determination of mercury by extraction and isotope-dilution mass spectrometry.

The use of halides for determination of metals by stable-isotope mass spectrometry is suggested, and is illustrated by the use of mercury iodide for mercury determination.

Photocontact allergy due to suprofen.

A 79-year-old Japanese woman developed edematous erythema on sun-exposed areas 3 months after applying Sulprotin ointment, which contains 1% suprofen (SP), a phenylacetic acid derivative. A patch test with Sulprotin ointment as it was negative, but a photopatch test with Sulprotein ointment as is was positive. A photopatch test with 10-3% SP was positive, as was that obtained with an SP analogue, 3% ketoprofen, but that with the ointment base of Sulprotin was negative. Photopatch tests with other SP analogues such as flurbiprofen, ibuprofen, and pranoprofen were negative. She developed abnormal erythema with monochromatic irradiation in the range from 320 to 380 nm on the uninvolved abdominal skin where 1% SP had been applied 24 hours (h) before irradiation. The action spectrum for this erythema agreed with the absorption spectrum of SP in the range of UVA. The MED after applying SP was below that after applying w. pet. in the range from 250 to 290 nm and from 300 to 320 nm. Positive intracutaneous test results were obtained with an irradiated mixture of human serum albumin (HSA) and SP at 48 and 72 h after injection and negative ones with a mixture of HSA and irradiated SP.

Actinic cheilitis granulomatosa.

A granulomatous lesion of the lips caused by UVB-photosensitivity is described. A 31-year-old Japanese man developed depigmented, swollen, erythematous lips with some erosions and bloody crusts which were present for 11 months. Histologic examination revealed a dense dermal infiltration composed mainly of lymphoid-histiocytic cells. Suprabasal clefts were associated with the invasion of lymphoid cells around them. Results of patch and photopatch testing were negative. The minimal erythema dose 24 h after irradiation was lower than normal, ranging from 295 to 330 nm. The lesion was reproduced by repeated irradiation with monochromatic light of 310 and 320 nm, but not by exposure to 250, 260, 270, 280, 290, 300, or 330 nm. Topical injection of glucocorticoids and application of sunscreen led to improvement. We propose the name "actinic cheilitis granulomatosa" for this case.

Adenocarcinoma with signet ring cells of the axilla: two case reports and review of the literature.

Adenocarcinoma with signet ring cells (ASRC) is a rare skin neoplasm whose histology shows a solid tumor intermingled with signet ring cells in variable numbers. There have been only ten reported cases. All were elderly males affected on the eyelids except for a single case in the axilla. Two new patients with ASRC of the axilla are described. In both of them, immunohistochemical studies revealed neoplastic cells that had differentiated toward apocrine glands. These are the second and third reported cases of ASRC in the axilla, one of them is the first ASRC case in a female. It seemed that the apocrine sweat gland or aberrant breast tissue in the axilla were possible origins of these tumors.

Livedo vasculitis associated with regional cytophagic histiocytoses.

We report a 70-year-old woman who developed recurrent multiple ulcers and residual atrophic scars on her legs and feet. The histology showed fibrinoid material in the dermal vessel walls causing partial to complete occlusion of the lumina. Capillary tufting and dilatation were also noted in the dermis. There was no vasculitis or panniculitis. Extravasated red blood cells were seen throughout the dermis; these were engulfed by benign-appearing histiocytes around the dermal-fat junction. As she was generally well and there was no evidence of the presence of cytophagic cells in her extracutaneous organs, she was diagnosed with regional phagocytic histiocytoses supposedly triggered by extravasated red blood cells.

The use of epoxy patch grafts for the repair of experimentally-created diaphragmatic defects in dogs.

Canine pericardium which had been treated with polyepoxy compounds (Denacol EX-313) was used as a patch graft for the correction of experimentally-created diaphragmatic defects in five dogs belonging to the same litter. Clinical, macroscopic and histological examinations were conducted every month up to five months after suturing of the patch graft. Clinical examination of the patch graft showed no apparent abnormalities. Macroscopic examination conducted during autopsy showed that the patch graft maintained adequate elasticity for five months after suturing, the surface of the patch graft was covered with a thin membrane and neovascularization was observed. Histological examination showed that the surface of the patch graft was covered with a thin membrane. Inflammatory tissue reactions were observed at one month, but gradually decreased from the second month onwards. In addition, the patch graft had excellent tissue affinity.

[Prevention of postoperative infection following cardiac catheterization in pediatric field. Study of air-borne bacteria in X-ray room and evaluation of an antibiotic used for prevention of postoperative infection].

As part of preventive measures against postoperative infection following cardiac catheterization in infants with cardiac diseases, especially falling bacteria in X-ray room was studied. Moreover, a synthetic penicillin, ticarcillin (TIPC), was used as preventive antibiotic against postoperative infections due to falling bacteria which probably contaminate the air in the X-ray examination room, and the efficacy and side effects of the drug were observed. As result, coagulase-negative Staphylococcus was detected the most, followed by Micrococcus and then by fungus. The number of these 3 organisms corresponded to 90.3% of the total number of falling bacteria detected during operation. The number of falling bacteria during operation was 5.1 times larger than that before operation. Taking into account normal flora of skin, falling bacteria present in the X-ray room and causative organisms of bacterial endocarditis, TIPC was administered to 30 cases intravenously 5 times at a dose of 30 mg/kg every 8 hours for the purpose of preventing possible postoperative infections following cardiac catheterization. The drug was effective to prevent such infections in all cases. No side effects were noted in any case, in peripheral blood and hepatic function tests and other observations.

[CD7+, CD34+, electronmicroscopically peroxidase-negative acute leukemia transformed from polycythemia vera after 12 years follow-up].

We reported a 72-year-old female patient who developed acute leukemia following a long course of polycythemia vera (PV). For 12 years she had been treated with phlebotomy, nimustine, busulfan, hydroxyurea and irradiation on splenomegaly. In November 1995, her peripheral blood smear showed blast of 30%. Bone marrow blasts were microscopically as well as electromicroscopically peroxidase-negative and CD7 and HLA-DR positive. Six months later, the blasts were positive for CD7, CD34 and HLA-DR. On the basis of morphologic, biochemical and immunophenotypic features, the patient was diagnosed acute leukemia, probably arising at a primitive multipotential stem cell level. She failed to respond to the various combination therapy including prednisolone, vincristine, cytarabine, daunorubicin and etoposide. The stem-cell-leukemia transformation in PV occurs rarely and may be refractory to chemotherapy.

[Hyperalgesia induced by intrathecal administration of nitroglycerin involves NMDA receptor activation in the spinal cord].

Spinal NMDA receptors are involved in hyperalgesia and chronic pain. The activation of spinal NMDA receptor results in the production of nitric oxide in the second order neurons in the spinal cord dorsal horn. We investigated the effects of intrathecally administered nitroglycerin (NTG) which releases nitric oxide in the cell. Formalin test which reflects phasic and tonic nociception was used as a nociceptive measure in rats with chronically implanted intrathecal catheters. Intrathecal injection of NTG resulted in the increase of flinching behavior induced by formalin injection to one paw in phase 1 (phasic) and phase 2 (tonic) responses in a dose-dependent manner. Intrathecally administered NMDA antagonist, MK-801 (MK) dose-dependently inhibited the effect of NTG but the effect was significant only in the phase 2 of the formalin test. MK given after formalin injection had significantly less effect on the phase 2 response. L-NAME (NOS inhibitor), MB (guanylate cyclase inhibitor) and HB (nitric oxide scavenger) significantly antagonized the hyperalgesic effect of NTG in the phase 2 of the formalin test. These results show that nitric oxide plays an important role in producing hyperalgesia in the spinal cord acting postsynaptically as well as pre-synaptically.

[The enhancement of formalin induced agitation behavior by intrathecal administration of prostaglandin E1].

The spinal action of pro-inflammatory agent, prostaglandin E1 (PGE1) was investigated in rats, using formalin test. Sprague-Dawley rats were implanted with chronic intrathecal catheters. PGE1 0.25 microgram or 2.5 micrograms injected intrathecally. Ten minutes later, 5% formalin 50 microliters was injected to the dorsum of one hindpaw. For the time-response analysis, the total number of flinches were counted and expressed as responses per minute for each rat. For the dose response analysis, the cumulative responses per minute over the first 5 min (phase 1) and the interval 10-60 min (phase 2) following the formalin injection were calculated for each rat. PGE1 2.5 micrograms produced a significant increase of flinching in both phase 1 and phase 2. Dose dependent increase of flinching was seen only in the phase 2. Touch evoked allodynia was observed after the injection of PGE1 2.5 micrograms, which lasted about 50 minutes. The enhanced flinches induced by intrathecal PGE1 were antagonized by the coadministration of MK-801, hemoglobin or methylene blue. Allodynia was antagonized only by coadministration of methylene blue. These results indicate that hyperalgesia induced by spinal PGE1 in both phase 1 and phase 2 of the formalin test involves an increased release of glutamate and the activation of NMDA receptor in the spinal cord. The mechanism of allodynia observed in the rats given high dose of PGE1 differs from that of hyperalgesia.

[Antitumor effect of S-1 and cisplatin treatment against human gastric cancer xenografted in nude mice].

The enhanced effects due to the combined use of oral administration of S-1 and intraperitoneal administration of Cisplatin (CDDP) were examined with gastric cancer xenografts (NUGC 4). S-1, a new anticancer drug, was daily administered at 10 mg/kg (qld x 5 x 3 weeks). 5-FU level in blood was 1 microgram/ml at two hours after the treatment. Antitumor activity was not found in mice with only the CDDP treatment. But antitumor activity by S-1 and daily low-dose (1 mg/kg) or intermittent treatment (5 mg/kg) of CDDP showed better results than daily S-1 treatment. The daily low-dose CDDP treatment showed similar efficacy to the intermittent administration at the same total dose, but the daily low-dose CDDP treatment was better in the light of toxicities. These results suggest that treatment with S-1 and daily low-dose CDDP was effective for gastric cancer.