Acute social defeat stress activated neurons project to the claustrum and basolateral amygdala.

We recently reported that a neuronal population in the claustrum (CLA) identified under exposure to psychological stressors plays a key role in stress response processing. Upon stress exposure, the main inputs to the CLA come from the basolateral amygdala (BLA); however, the upstream brain regions that potentially regulate both the CLA and BLA during stressful experiences remain unclear. Here by combining activity-dependent viral retrograde labeling with whole brain imaging, we analyzed neurons projecting to the CLA and BLA activated by exposure to social defeat stress. The labeled CLA projecting neurons were mostly ipsilateral, excluding the prefrontal cortices, which had a distinctly labeled population in the contralateral hemisphere. Similarly, the labeled BLA projecting neurons were predominantly ipsilateral, aside from the BLA in the opposite hemisphere, which also had a notably labeled population. Moreover, we found co-labeled double-projecting single neurons in multiple brain regions such as the ipsilateral ectorhinal/perirhinal cortex, entorhinal cortex, and the contralateral BLA. These results suggest that CLA and BLA receive inputs from neuron collaterals in various brain regions during stress, which may regulate the CLA and BLA forming in a stress response circuitry.

Determination of O:4 antigen-antibody affinity level in O:5 antigen positive and negative variants of Salmonella enterica serovar Typhimurium.

Salmonella enterica serovar Typhimurium (S. Typhimurium) has two serological variants: one that expresses the O:5 antigen (1,4,5,12:i:1,2) and one that lacks O:5 antigen (1,4,12:i:1,2). For serotyping, S. Typhimurium is agglutinated by diagnostic O:4 antigen serum. This study was carried out to compare the antigen-antibody affinity of O:4 antigen in S. Typhimurium χ3306 O:5-positive and S. Typhimurium χ3306 O:5-negative strains. The affinity of O:4 antigen with O:4 antigen serum was found to be stronger in the O:5-negative strains compared to O:5-positive strains. Next, we investigated the antigen-antibody affinity of O:4 antigen with O:4 antigen serum in field strains of S. Typhimurium, which showed the same tendency in affinity as seen with S. Typhimurium χ3306 O:5-positive and negative strains. This study suggests that the presence or absence of O:5 antigen causes differences in O:4 agglutination reactions with different field strains of S. Typhimurium.

The Role of an Impurity in Ceftriaxone Sodium Preparation for Injection in Determining Compatibility with Calcium-Containing Solutions.

Ceftriaxone sodium preparation for injection is known to form insoluble microparticles with calcium. The purpose of this study was to evaluate the role of an impurity in the ceftriaxone sodium preparation on this incompatibility. Firstly, using HPLC, two impurities were identified in the ceftriaxone sodium solution. The major impurity (impurity 1) was identified as tetrahydro-2-methyl-3-thioxo-1,2,4-triazine-5,6-dione by LC/MS. Secondly, the role played by this impurity in the incompatibility with calcium was examined. Using seven different ceftriaxone preparations for injection, the effect of adding impurity 1 to mixed solutions of ceftriaxone sodium and calcium chloride on the appearance of insoluble microparticles, was examined using a light obscuration particle counter. Although incompatibility was not completely suppressed by the addition of impurity 1, the number of insoluble microparticles formed with calcium chloride solution was decreased in proportion to the concentration of impurity 1, and the concentration of calcium ion decreased as the concentration of added impurity 1 increased. These results show that impurity 1 plays a concentration-dependent role in incompatibility between ceftriaxone sodium preparation for injection and calcium-containing solutions.

[A Case of a Suspected Photosensitive Reaction Induced by Weekly Paclitaxel as Neoadjuvant Chemotherapy for Breast Cancer].

We report a suspected photosensitive reaction induced by weekly paclitaxel (PTX) in a patient with breast cancer. The patient was a 72-year-old woman with right breast cancer (T2N1M0, Stage II B). She received 80 mg/m² PTX weekly as neoadjuvant chemotherapy. After the 5 courses of weekly PTX, she began to develop skin lesions with itchy sensations on the areas of her body that had been exposed to sunlight. The symptoms persisted after the 6 courses of weekly PTX, and topical steroid and antihistamine treatments were initiated. The skin lesions resolved once the patient avoided sunlight and used sunscreen. Ultimately, she received 12 courses of PTX. We need to recognize the appearance of a photosensitive reaction following weekly PTX for breast cancer.

Prediction of compatibility between ozagrel sodium preparation for injection and calcium on the basis of the solubility product.

The purpose of the study was to evaluate the compatibility of ozagrel sodium solution and calcium-containing transfusions using solubility product constants. We calculated the solubility product constant of mixtures of ozagrel sodium and calcium chloride and evaluated the compatibility of ozagrel sodium solution (both the original and generic products) with calcium chloride solution using a light obscuration particle counter. Various volumes of ozagrel solution were added to the calcium solutions to make final ozagrel concentrations of 0, 0.8, 1.6, 2.0, 2.4, 3.2 and 4.0 mmol/L. The solutions were gently agitated and stored at 25 and 40°C. The ozagrel concentration, calcium ion concentration and number of microparticles were measured. The solubility product constants obtained were 11.89×10(-9) mol(3)/L(3) (at 25°C) and 7.82×10(-9) mol(3)/L(3) (40°C). The number of insoluble microparticles was significantly increased when the ionic product was larger than the solubility product constant. In all ozagrel sodium products, the number of insoluble microparticles was within the allowable range according to the Japanese Pharmacopoeia. These results suggest that mixing ozagrel sodium with calcium-containing products is safe and without appreciable risk of incompatibility under clinical conditions.

Comparison between original and generic versions of ceftriaxone sodium preparation for injection: compatibility with calcium-containing product.

The purpose of this study was to compare the compatibility of ROCEPHINE® Intravenous, the original manufacturer's ceftriaxone sodium preparation for injection, and seven generic versions thereof, with various calcium chloride injection 2%. The influence of calcium ion concentration, storage time and shaking strength on the appearance and quantity of insoluble microparticles in mixed solutions was examined using a light obscuration particle counter. In all products, the observed number of insoluble microparticles was proportional to the calcium ion concentration, storage time and shaking strength after the addition of calcium chloride solution. In several of the generic products, the number of insoluble microparticles was significantly higher than those of the original product, while in others it was lower. We evaluated the quality of the original and 7 generic preparations, measured the content of impurity and pH of the various ceftriaxone solutions, as impurity content and pH of solution are possible factor affecting compatibility. Three impurities were found in all products. The impurity content of several generic products, as estimated from their peak area on high performance liquid chromatography (HPLC), was significantly lower than that of the original product. pH of solution was difference between products. Although it was difficult that impurity and pH of solution verify critical factor affecting compatibility. The results show that there are differences in the appearance of insoluble microparticles between the original product and seven generic products, when calcium chloride injection 2% solution is added to the product.

Mechanisms of resistance to cephalosporin and emergence of O25b-ST131 clone harboring CTX-M-27 ß-lactamase in extraintestinal pathogenic Escherichia coli from dogs and cats in Japan.

Thirty-three cefazolin-resistant extraintestinal pathogenic Escherichia coli strains from companion animals were screened for bla(CMY-1) , bla(CMY-2) , bla(SHV) , bla(TEM) , and bla(CTX-M) genes. Extended-spectrum ß-lactamase-producing strains were further characterized by O serotyping and multilocus sequence typing. It was found that 20 and 17 isolates harbored TEM-1 and CMY-2 ß-lactamases, respectively, and 13 isolates harbored both ß-lactamases. One isolate harbored DHA-1 ß-lactamase. Eleven isolates were found to possess CTX-M ß-lactamases (CTX-M-27 [n= 6], CTX-M-14 [n= 3], CTX-M-15 [n= 1], and CTX-M-55 [n= 1]). Of 11 CTX-M-positive strains, four strains were O25b-ST131 clones harboring CTX-M-27, and the remaining seven strains belonged to O6-ST127, ONT-ST354, O159-ST539, O1-ST648, O8-ST1642, O25b-ST2042, and ONT-ST2178.

Prevalence of antimicrobial resistance in relation to virulence genes and phylogenetic origins among urogenital Escherichia coli isolates from dogs and cats in Japan.

OBJECTIVE: To assess the status of antimicrobial resistance (AMR), identify extraintestinal virulence factors (VFs) and phylogenetic origins, and analyze relationships among these traits in extraintestinal pathogenic Escherichia coli (ExPEC) isolates from companion animals. SAMPLE: 104 E coli isolates obtained from urine or genital swab samples collected between 2003 and 2010 from 85 dogs and 19 cats with urogenital infections in Japan. PROCEDURES: Antimicrobial susceptibility of isolates was determined by use of the agar dilution method; a multiplex PCR assay was used for VF gene detection and phylogenetic group assessment. Genetic diversity was evaluated via randomly amplified polymorphic DNA analysis. RESULTS: Of the 104 isolates, 45 (43.3%) were resistant to > 2 antimicrobials. Phylogenetically, 64 (61.5%), 22 (21.2%), 13 (12.5%), and 5 (4.8%) isolates belonged to groups B2, D, B1, and A, respectively. Compared with other groups, group B2 isolates were less resistant to all tested antimicrobials and carried the pap, hly, and cnf genes with higher frequency and the aer gene with lower frequency. The aer gene was directly associated and the pap, sfa, hly, and cnf genes were inversely associated with AMR. Randomly amplified polymorphic DNA analysis revealed 3 major clusters, comprised mainly of group B1, B2, and D isolates; 2 subclusters of group B2 isolates had different VF and AMR status. CONCLUSIONS AND CLINICAL RELEVANCE; Prevalences of multidrug resistance and human-like phylogenetic origins among ExPEC isolates from companion animals in Japan were high. It is suggested that VFs, phylogenetic origins, and genetic diversity are significantly associated with AMR in ExPEC.

[Efforts of pharmacists in regimen management for safe cancer chemotherapy].

As a safe way to manage cancer chemotherapy, a regimen registration system of chemotherapies was started in our hospital in April of 2007. Only chemotherapies registered in the regimen system can be conducted. Beginning in July of 2007, to assure safer chemotherapy, a standard clinical value of laboratory data was established as the criteria to discontinue chemotherapies. The individual laboratory data have been checked according to the discontinuance criteria by pharmacists in our hospital. With the adoption of this regimen registration system for standardized procedures of various chemotherapies, laboratory data can be checked according to the criteria, and certain chemotherapies can now be discontinued. However, in certain cases some doctors proceeded chemotherapies although pharmacists suggested that laboratory data did not meet the criteria. It is important to keep checking the registered regimen and criteria. Pharmacists have to confirm the checking system based on the patient's condition and improve the regimen system of chemotherapy more clearly together with doctors and nurses.

Prediction of incompatibility of ceftriaxone sodium with calcium ions using the ionic product.

The purpose of this study was to evaluate the incompatibility of ceftriaxone sodium with calcium-containing products using the ionic product of precipitation, and the measurement of insoluble microparticles using a light obscuration particle counter. Appropriate volumes of 2% (w/v) calcium chloride solution were added to 0.4-2 mg/ml ceftriaxone isotonic sodium chloride solution, to make solutions with a final calcium ion concentration of 1.25 mmol/l. The solutions were gently agitated and stored at 37 degrees C for 24 h. The number of insoluble microparticles with a diameter less than 10 microm in the mixed sample solution, determined using a light obscuration particle counter, was increased when the ceftriaxone concentration was > or =0.8 mg/ml. The Saturation Index (defined as the ratio of the ionic product to the solubility product constant) of the prepared mixed solution was 1.1. A white precipitate could be observed visually when the ceftriaxone concentration of the sample solution was 7 mg/ml; the Saturation Index of the solution was 9.7. The effect of the calcium source on incompatibility with ceftriaxone sodium was also evaluated. The numbers of insoluble microparticles in sample solutions made by adding calcium chloride to the sample were significantly higher than those made by adding calcium gluconate. These results suggest that ceftriaxone should not be co-administered with calcium-containing products even if no precipitation is observed visually. There will still be insoluble microparticles caused by incompatibility in the sample solution when the Saturation Index of the solution is over 1.0.

Incompatibility of ceftriaxone sodium with calcium-containing products.

The purpose of this study was to evaluate the incompatibility of ceftriaxone with calcium-containing products, which had been the subject of an ALERT issued by the FDA. The influence of calcium ion concentration, storage temperature and shaking on the appearance and quantity of insoluble microparticles in mixtures of the two was examined using a light obscuration particle counter and a stereomicroscope. Appropriate volumes of 2% (w/v) calcium chloride solution were added to ceftriaxone sodium for injection (10 mg/ml) to make solutions with final calcium ion concentrations of 0.5-2.5 mmol/l, and stored at 20 degrees C, 25 degrees C, or 30 degrees C. The number of insoluble microparticles increased on storage when the calcium ion concentration of the sample was > or =2 mmol/l; it exceeded the permissible range at all temperatures by 1 h. The microparticles had a greater diameter at higher temperatures, although fewer microparticles were observed. The weight of precipitate increased as a function of both the calcium ion concentration and the temperature. The number of microparticles was also significantly increased by shaking. The number of microparticles in mixtures containing 1000 microg/ml ceftriaxone was significantly increased, even though concentrations of calcium ion was 1.25 mmol/l. Overall, not only calcium ion concentration, but storage temperature and shaking affected the extent of precipitation of ceftriaxone with calcium.

Enhancement of gene transfection into human dendritic cells using cationic PLGA nanospheres with a synthesized nuclear localization signal.

Effective delivery of DNA encoding antigen into the dendritic cells (DCs), which are non-dividing cells, is very important for the development of DNA vaccines. In a previous study, we developed the PLGA nanospheres that contained a cationic nanomaterial and showed high transfection efficiency in COS7 cells, which divide. In the present study, to produce an effective vector for the DNA vaccines, the gene expression and intracellular trafficking of pDNA complexed with PLGA/PEI nanospheres, in combination with an NF-kappaB analog as a nuclear localization signal (NLS) and electroporation were evaluated in human monocyte-derived DCs (hMoDCs). Cellular uptake of pDNA both in COS7 cells and hMoDCs was enhanced using the PLGA/PEI nanospheres. On the other hand, the PLGA/PEI nanospheres significantly promoted the transfection in COS7 cells, but had almost no effect on transfection in hMoDCs. The intranuclear transport of pDNA by PLGA/PEI nanospheres in COS7 cells was significantly higher than that in hMoDCs. These results indicate that pDNA complexed with PLGA/PEI nanospheres cannot enter into the nuclei of non-dividing cells. However, PLGA/PEI nanospheres combinated with NLS and electroporation (experimental permeation enhancer) greatly elevated the transfection efficiency by improvement of not only intracellular uptake but also intranuclear transport of pDNA in the hMoDCs. Thus, this delivery system using nanospheres combined with synthesized NLS might be applicable to DC-based gene vaccines when much non-invasive application such as needle-free injector should be required.

Quantitative taste evaluation of total enteral nutrients.

The purpose of this study was to evaluate quantitatively the taste of the various total enteral nutrients marketed in Japan using human gustatory sensation tests and an artificial taste sensor. In the human gustatory sensation test, four basic taste intensities (sweetness, saltiness, sourness, and bitterness), as well as 15 kinds of palatability scales, were evaluated according to the semantic differential (SD) method. Among 15 palatability items, the item; difficult to drink/easy to drink, was adopted as an overall palatability since it shows the highest factor loading by factor analysis. The overall palatability was found to be highly positively correlated with sweetness and sourness, but negatively correlated with bitterness and saltiness. Addition of a flavour to the amino acid-based enteral nutrient AminolebanEN significantly improved its palatability. This effect is presumably due to sour components of the flavour, such as citric acid, which reduce the bitterness intensity of branched-chain amino acids in the product. The sweetness and sourness intensities predicted by the taste sensor showed a high correlation with the results obtained in the human gustatory sensation tests. The taste sensor was able to predict the overall palatability of the total enteral nutrients with high accuracy. The products could be classified into three groups (peptide-based, amino-acid-based, and protein-based) by principal component analysis using sensor output of 8 channels. The products could be also classified into four groups; peptide-based, amino-acid-based, and protein-based and flavor addition group by principal component analysis using sensor output of channels 1, 3, 4 and 7, which are specific to basic tastes. The taste sensor could therefore be useful in predicting the taste or palatability of total enteral nutrients, and could contribute to attempts to improve compliance for such products and for enteral nutrients.

Bitterness evaluation of medicines for pediatric use by a taste sensor.

The purpose of this study was to evaluate the bitterness of 18 different antibiotic and antiviral drug formulations, widely used to treat infectious diseases in children and infants, in human gustatory sensation tests and using an artificial taste sensor. Seven of the formulations were found to have a bitterness intensity exceeding 1.0 in gustatory sensation tests (evaluated against quinine as a standard) and were therefore assumed to have an unpleasant taste to children. The bitterness intensity scores of the medicines were examined using suspensions in water or an acidic sports drink. In the case of three macrolide antibiotic formulations containing erythromycin (ERYTHROCIN dry syrup), clarithromycin (CLARITH dry syrup for pediatric), and azithromycin (ZITHROMAC fine granules for pediatric use), the bitterness intensities of suspensions in acidic sports drinks were dramatically enhanced compared with the corresponding scores of suspensions in water. This enhancement could be predicted using the taste sensor. On the other hand, a reduction of bitterness intensity was observed for an acidic sports drink suspension of an amantadine product (SYMMETREL fine granules) compared with an aqueous suspension. This reduction in bitterness could also be predicted using the taste sensor output value. Thus, the taste sensor could predict whether or not suspension in an acidic sports drink would enhance or reduce the bitterness intensity of pediatric drug formulations, compared with suspensions in water.

Case of adrenal lymphangioma with atypical lymphocytes in aspirate cytology.

Cytological diagnosis of adrenal cysts is becoming more important, with the increase in cases identified due to the widespread use of imaging modalities. Adrenal cysts have been classified into four main groups: endothelial, epithelial, parasitic, and pseudocyst, with endothelial cysts further classified as lymphangiomatous or angiomatous. We report on a case of adrenal lymphangioma with atypical lymphocytes found in the intraoperative aspirate, suggesting an inconclusive diagnosis, although immunohistochemical study led us to suppose their B-cell origin. This case provided important cytological findings that may assist in the prevention of cytological overdiagnosis.