Prevalence of intestinal parasites and genotyping of Giardia intestinalis in pet shop puppies in east Japan.

The current study examined the prevalence of intestinal parasites and genotypes of Giardia intestinalis in puppies from nine pet shops in east Japan. Fresh fecal samples from 1794 puppies (≦3 months old) were collected on one occasion. Giardia spp. was examined for specific coproantigen using ELISA kit (SNAP®Giardia, IDEXX Laboratories, Inc., USA). Other intestinal parasites were detected microscopically using the formalin-ethyl acetate sedimentation technique. Genotyping was determined for the random 29 stool samples identified as Giardia spp. positive using PCR and direct sequencing of the glutamate dehydrogenase (gdh) gene. Overall prevalence of protozoan Giardia spp. and Cystoisospora spp. revealed 23.4% and 11.3%, respectively. Prevalence of ascarids, Strongyloides spp. and hookworms were recorded 1.8%, 1.1% and 0.1%, respectively. Protozoan Giardia spp. and Cystoisospora spp., thus, represent important pathogens among pet shop puppies. All genotyped G. intestinalis isolates were belonged to assemblage C or D, identified as dog-specific genotypes. Zoonotic assemblage A and B were not demonstrated. The result suggests that the risk of zoonotic transmission of G. intestinalis from pet shops puppies to humans may be quite low in Japan.

[Carinal resection of bronchogenic carcinoma].

Carinal resection was performed in 10 cases of bronchogenic carcinoma during 12 year-period. The mean age of patients was 58 years, with a range of 42 to 68 years. There were 7 male and 3 female. The tumor was located on the right side in 7 cases, on the left side in 1 case on the carina in 2 cases. The histological examination showed squamous cell carcinoma in 5 cases, adenocarcinoma in 4 cases and adenoid cystic carcinoma in 1 case. The staging revealed T3N2 M0 Stage IIIA in 2 cases, T4N0M0 Stage IIIB in 1 case, T4N1M0 Stage IIIB in 1 case, T4N2M0 Stage IIIB in 5 cases and T4N3M0 Stage IIIB in 1 case. The surgical methods were as follows; sleeve pneumonectomy in 5 cases, wedge carinal resection with pulmonary resection in 3 cases (right sleeve upper lobectomy in 2 cases and right pneumonectomy in 1 case), carinal resection in 2 cases. The site of bronchial anastomosis was overlapped by thymus in 6 cases. The 30-day mortality rate in tracheo-carinal resection was 10% (one patient). Eight patients died and remaining 2 patients are still alive without any evidence of recurrence. 5-year survival rate was 36%. These outcomes were almost equal to those of surgical case in the same stage.

Properties of the bimodal fluorescent protein produced by Photobacterium phosphoreum.

A fluorescent protein isolated from the deep-sea luminous bacterium Photobacterium phosphoreum strain bmFP has been purified, cloned and sequenced. The protein is 96.5% identical in amino acid sequence to FP390, the weakly fluorescent flavoprotein encoded by the luxF gene characteristic of Photobacterium species. Similar to FP390, bmFP is a dimer of two homologous subunits binding four FMN-myristate chromophores but has the distinctive feature of emitting a bimodal fluorescence with maxima at about 488 and 517 nm, hence the name bmFP. For both bands of this fluorescence, the excitation spectrum exhibits a peak at 336 nm, not corresponding to its flavin-like absorption spectrum. Heating of bmFP in urea resulted in a decrease in the intensity of the 488 nm band along with the appearance of a new fluorescence peaking at 423 nm, partially reversible upon the removal of the urea. Upon complete denaturation, either by heat or guanidium chloride at 65 degrees C, fluorescence characteristic of both free flavin and this 423 nm species appears. It is speculated that chromophores in different states of protonation, associated with a single protein, are responsible for the unusual spectral properties of bmFP.

Activities of the bimodal fluorescent protein produced by Photobacterium phosphoreum strain bmFP in the luciferase reaction in vitro.

The activity of the bimodal fluorescent protein (bmFP) (lambda max, 488 and 517 nm) in the in vitro luciferase reaction has been studied. The bmFP that is produced by Photobacterium phosphoreum strain bmFP is a dimer of two homologous subunits binding four riboflavin 5'-phosphate (FMN)-myristate chromophores. The addition of bmFP to the luciferase reaction in the presence of the lumazine protein prevented the lumazine protein-induced blue shift in the emission band. The bmFP reduced electrochemically serves as a substrate in the luciferase reaction in the absence of added FMN, resulting in light emission with a single maximum at about 487 nm. The bmFP was also active in lieu of FMN in the NADH/FMN oxidoreductase (flavin reductase)-luciferase coupled bioluminescence reaction in the absence of added FMN. In the coupled reaction, bioluminescence with the isolated bmFP chromophore was weaker than that with the holo-bmFP. After bmFP was used in luciferase reactions initiated either chemically or electrochemically, it was still capable of emitting bimodal fluorescence.