New options of antibiotic combination therapy for multidrug-resistant Pseudomonas aeruginosa.

Several antibiotic combinations have demonstrated increased activity against multidrug-resistant Pseudomonas aeruginosa (MDRP) in vitro compared with a single antibiotic. The aim of this study was to investigate the activity against MDRP of some aminoglycosides in combination with monobactam, piperacillin (PIPC), and carbapenem. Clinical isolates of MDRP were collected between November 2010 and October 2012 from patients in Tokyo Medical University Hospital, Tokyo (1,015 beds). Our new method was designed to evaluate three concentrations around the breakpoint of each drug using the Checkerboard method. The aminoglycosides tested were amikacin (AMK), tobramycin (TOB), and arbekacin (ABK). Ciprofloxacin, PIPC, and biapenem (BIPM), which have been reported to demonstrate combination effects, were also tested. Sixty-six MDRP strains were identified from the 2,417 P. aeruginosa strains. Of the 66, 27 tested positive for metallo-ß-lactamase (MBL). Aztreonam (AZT) with AMK or ABK was the most effective against MDRP. PIPC with AMK or ABK were somewhat effective. AZT with AMK or ABK were more effective against MBL-positive strains than MBL-negative strains. However, PIPC with AMK or ABK were more effective against MBL-negative strains than MBL-positive strains. Combination activities showed differences between MBL-positive and MBL-negative strains.

Preclinical anticancer effects and toxicologic assessment of hepatic artery infusion of fine-powder cisplatin with lipiodol in vivo.

We conducted an in vivo study to evaluate the anticancer effect and toxicity of fine-powder cisplatin suspended in lipiodol (fCDDP/LPD suspension) after a single administration of three different doses to rats via the intrahepatic artery after transplantation of rat ascites hepatoma cells. The toxicity of the fCDDP/LPD suspension was also assessed in the same protocol in noncancer-bearing rats and the observed toxicologic changes were compared among groups administered saline (Sal), an aqueous solution of fCDDP (fCDDP/Sal solution), and LPD alone. In parallel with the toxicity test, plasma CDDP concentrations were compared between the fCDDP/LPD suspension and fCDDP/Sal solution. The mean weight of the tumors in the fCDDP/LPD suspension groups was significantly less than in the LPD-alone group. The pathologic changes in the liver observed in the fCDDP/LPD suspension group increased with dose, were more marked compared with those in the fCDDP/Sal solution and LPD-alone groups, and were reversible. No other toxicologic effects were observed. The concentration of CDDP in the plasma in the fCDDP/LPD suspension group was slightly lower than that in the fCDDP/Sal solution group. In conclusion, the results indicate that the fCDDP/LPD suspension has sufficient anticancer efficacy and tolerability for use in the clinical treatment of hepatocellular carcinoma.

Fatal Aspergillus pancarditis after incompatible blood transfusion intended to be an autologous blood transfusion.

Aspergillus pancarditis is a rare infection, and it has rarely been reported after blood transfusion. In this report, we describe a fatal case of Aspergillus pancarditis in a patient who received antibiotics and corticoids after an incompatible blood transfusion intended to be an autologous blood transfusion. A 64-year-old man suffering from herniation of intervertebral disk between C4 and C5 received an anterior cervical spinal fusion. After the operation, he received incompatible blood transfusion and fell into disseminated intravascular coagulation (DIC) and preshock state. Anticoagulants and corticoids were given and he recovered from DIC and the preshock state. However, he remained quadriplegic because of enlargement of cervical epidural hematoma that was initially brought out by the operation. He developed bacterial bronchopneumonia 2 weeks after the transfusion and received antibiotic therapy. The pneumonia was cured a week later. Five weeks after the transfusion, he developed Aspergillus pneumonia and received antimycotic therapy. However, his condition grew worse and died 2 months after the transfusion. Autopsy revealed Aspergillus pancarditis. In this case, the relationship between the erroneous transfusion and the patient's death was obvious and it was considered that the erroneous transfusion should be blamed for the patient's death.

Discrimination among three species of medicinal Scutellaria plants using RAPD markers.

An analysis of random amplified polymorphic DNA (RAPD) was performed using nine accessions of three species of medicinal plants in the genus Scutellaria (S. galericulata, S. lateriflora and S. baicalensis; known collectively as skullcap) in an effort to distinguish between members of these three species. Dried aerial parts of the two species S. galericulata and S. lateriflora are difficult to distinguish morphologically. Ten arbitrary primers produced 92 fragments, and eight of the primers yielded 23 species-specific fragments among the three species. Six fragments were specific for S. galericulata, seven for S. lateriflora and ten for S. baicalensis. When primers A02 and A06 were used in the polymerase chain reaction, RAPD fragments that were specific for each of the three species were generated simultaneously. Primer A02 produced five species-specific fragments: one was specific for S. galericulata; two for S. lateriflora; and two for S. baicalensis. Primer A06 produced three species-specific fragments: one for S. galericulata; one for S. lateriflora; and one for S. baicalensis. The RAPD markers that were generated with these two primers should rapidly identify members of the three species of Scutellaria. The consistency of the identifications made with these species-specific RAPD markers was demonstrated by the observation that each respective marker was generated from three accessions of each species, all with different origins. Furthermore, cluster analysis using the 92 RAPD fragments produced a dendrogram of genetic relatedness that was in good agreement with the taxonomic designations of the three species. Thus, the RAPD markers should be useful for the future identification of members of the three species of medicinal Scutellaria plants.

Malignant rhabdoid tumor of the colon: report of a case.

A malignant rhabdoid tumor of the colon is very rare and only three cases have been previously described. A 76-year-old man was admitted to the hospital complaining of epigastralgia. An elastic mass was palpable in the right upper abdomen. A barium enema and endoscopic examination showed a giant gyrate tumor arising from the cecum. Abdominal ultrasonography and a computed tomography scan revealed the tumor to be located in the colon associated with multiple liver metastases and gallbladder stones. A right colectomy and cholecystectomy were thus performed. The tumor was histologically composed of sheets of large round and polygonal nuclei with vesicular chromatin, and abundant acidophilic cytoplasm, often containing hyalin-like inclusion. The cytoplasm was positive for vimentin and neuron-specific enolase, and hyaline globules of the rhabdoid tumor cells stained positive for cytokeratin in some cells. Transmission electron microscopy showed characteristic rhabdoid cells with an aggregation of intermediate filaments. A histologic diagnosis of malignant rhabdoid tumor of the colon was made. The tumor demonstrated several unusual findings for malignant rhabdoid tumors including diploidy by a flow cytometric analysis, and positive nuclear immunohistochemical staining for p53 protein and Ki-67 antigen. We report herein the third known case of a pure colonic rhabdoid tumor.

Sensitive high-performance liquid chromatographic determination of nifedipine in cat plasma following improved sample treatment.

A simple, easy and accurate reversed-phase high-performance liquid chromatographic method is described for the determination of nifedipine in cat plasma. The procedure involves extraction of nifedipine from plasma using a Sep-Pak C18 cartridge and ultraviolet detection at 350 nm. The present method provides the required reproducibility and sensitivity for the determination of low concentrations of nifedipine without interference from plasma components or photodegradation products. The method was validated over the range 1-50 ng/ml nifedipine. Accuracy and precision were, respectively, 97% or more and 5% or less over the concentration range examined. The minimum quantifiable concentration of nifedipine was found to be 1 ng/ml.

Mutation in Npps in a mouse model of ossification of the posterior longitudinal ligament of the spine.

Ossification of the posterior longitudinal ligament of the spine (OPLL) is a common form of human myelopathy caused by a compression of the spinal cord by ectopic ossification of spinal ligaments. To elucidate the genetic basis for OPLL, we have been studying the ttw (tiptoe walking; previously designated twy) mouse, a naturally occurring mutant which exhibits ossification of the spinal ligaments very similar to human OPLL (refs 3,4). Using a positional candidate-gene approach, we determined the ttw phenotype is caused by a nonsense mutation (glycine 568 to stop) in the Npps gene which encodes nucleotide pyrophosphatase. This enzyme regulates soft-tissue calcification and bone mineralization by producing inorganic pyrophosphate, a major inhibitor of calcification. The accelerated bone formation characteristic of ttw mice is likely to result from dysfunction of NPPS caused by predicted truncation of the gene product, resulting in the loss of more than one-third of the native protein. Our results may lead to novel insights into the mechanism of ectopic ossification and the aetiology of human OPLL.

Sensitization of nociceptive C-fibers in zinc-deficient rats.

A marked decrease in zinc concentration was observed in plasma (P < 0.001), hindpaw skin (P < 0.01), and dorsal skin (P < 0.01) in zinc-deficient rats (rats fed a zinc-deficient diet for 3 wk), compared with the control rats fed the same zinc-deficient diet supplemented with ZnCO3 (50 mg/kg diet). The threshold intensity needed to elicit vasodilatation in the hindpaw skin of the zinc-deficient rats on electrical stimulation of the saphenous nerve in a peripheral direction was markedly lower (P < 0.01) than that in the control rats. No difference was observed between control (n = 5) and zinc-deficient rats (n = 5) in the magnitude of the plasma extravasation evoked by either histamine or substance P. There was no difference between control and zinc-deficient rats in terms of the dose-response curve for release of histamine by substance P. Prostaglandin E2 (PGE2) concentration in the hindpaw skin of the zinc-deficient rats was nearly fourfold higher (P < 0.01) than that of the control rats, whereas no difference in the leukotriene B4 level in the hindpaw skin was observed between control and zinc-deficient rats. From the present study, it seems likely that an increased level of PGE2 in the vicinity of the nociceptive C-fiber terminals in the hindpaw skin of zinc-deficient rats may sensitize the terminals of the nociceptive C-fibers of the saphenous afferent nerve in the hindpaw and thus facilitate the production of antidromic vasodilatation.

Expression of novel DNA-binding protein with zinc finger structure in various tumor cells.

We have isolated from B16 mouse melanoma cells a complementary DNA (Mel-18), whose deduced amino acid sequence possesses a characteristic zinc finger structure. Immunostaining with antibodies raised against partial Mel-18 peptide sequences demonstrated nuclear localization of the gene product. We have also demonstrated that this protein has DNA-binding capacity, and the zinc finger is responsible for the DNA binding. At the transcriptional level the Mel-18 mRNA was detected in all tumor cells examined as well as melanoma cells (ontogenically of neural origin) but was scarcely present in normal tissues except neural organs. The transcript is developmentally regulated. These data suggest that Mel-18 may play a role in transcriptional regulation and also in control of cell proliferation and/or neural cell development.

[Morphometric study of the magnocellular basal forebrain system in patients with Alzheimer's disease].

Morphometric study of neurons within the magnocellular basal forebrain system (MBFS) in the three normal controls and three cases with Alzheimer's disease (AD) was studied. Immunocytochemical staining using anti-acetylcholinesterase was performed to identify the measuring areas of the MBFS, and cross sectional areas of all neurons within the MBFS in one preparation was measured making use of cresyl violet staining. About 50% of the entire neuronal cells within the MBFS were decreased and case 2 had neurofibrillary tangles in the substantia innominata. In the septal nucleus neuronal cell depopulation was observed through the all range of the neuronal cell size, and in the diagonal band of Broca neuronal cells of which cross sectional areas were more than 200 microns2 were preferentially decreased and case 3 had inverse increase of the neurons of which cross sectional areas were less than 200 microns2. In the basal nucleus of Meynert in the substantia innominata neurons of which cross sectional areas were more than 250 microns2 were markedly decreased and neurons of which cross sectional areas less than 250 microns2 were well preserved. The large neurons within the basal nucleus and diagonal band of Broca were more affected in AD. In the septal nucleus and diagonal band it was suspected that non-cholinergic neurons were also decreased and these findings suggested that other series of monoamines also bore a relationship to the dementia and neuropsychological symptoms in AD.

Intratumor chemotherapy in combination with a systemic antimetastatic drug in the treatment of Lewis-lung carcinoma.

The effect of an antimetastatic agent plus intratumor chemotherapy was evaluated in mice bearing Lewis-lung carcinoma by measuring survival time and by histological examination. Polymeric flavan-3,4-diol (APF) from avocado seeds, Persea gratissima, administered alone directly into the tumor did not change survival time, although it partially destroyed the primary tumor. However, the drug administered in combination with an antimetastatic, 1,2-bis(3,5-dioxopiperazin-1-yl)ethane (ICRF-154), resulted in an increase in survival time. When 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) was used in place of polymeric flavanadiol as an intralesional drug, a significant increase in survival was also achieved. The effect of each drug alone and of their combination was evaluated by "responder analyses". Animals "cured" by the combination and rechallenged with 2 X 10(6) tumor cells showed that immunization could occur.