Persistent organic pollutants in red-crowned cranes (Grus japonensis) from Hokkaido, Japan.

The red-crowned crane (Grus japonensis) from eastern Hokkaido is classified as a Special Natural Monument in Japan. In this study, we determined the concentrations of persistent organic pollutants (POPs) in red-crowned crane muscle tissues (n = 47). Polychlorinated biphenyls (PCBs) had the highest median concentration (240ng/g lipid weight), followed by dichlorodiphenyltrichloroethane and its metabolites (DDTs) (150ng/g lipid weight), chlordane-related compounds (CHLs) (36ng/g lipid weight), hexachlorobenzene (HCB) (16ng/g lipid weight), hexachlorocyclohexanes (HCHs) (4.4ng/g lipid weight), polybrominated diphenyl ethers (PBDEs) (1.8ng/g lipid weight), and finally, Mirex (1.5ng/g lipid weight). Additionally, a positive correlation was found among POP concentrations. No sex differences beyond body parameters were observed. Additionally, red-crowned cranes exhibited a high enantiomeric excess of (+)-alpha-HCH, with enantiomer fractions varying from 0.51 to 0.87 (average: 0.69).

Low Genetic Variation of Red-Crowned Cranes on Hokkaido Island, Japan, Over the Hundred Years.

The red-crowned crane (Grus japonensis) is recognized internationally as an endangered species. Migratory populations breed in eastern Russia and northeastern China, whereas the resident population inhabits the island of Hokkaido, Japan. Although the population inhabiting Hokkaido had experienced a severe bottleneck by the end of the 19th century, the population size has recovered to about 1500 and continues to increase now thanks to conservation efforts. A previous study reported that no marked genetic differences were seen in the island population, and that the genetic variation of the whole population on Hokkaido was lower than that of the continental population. However, the precise genetic structure of the island population in the past or near present remains unclear. To better understand the spatiotemporal changes in the genetic structure of the island population, we performed mitochondrial DNA (mtDNA) analyses using stuffed specimens (years 1878-2001) and tissue or blood samples (years 1970-2014). We found three haplotypes in the island population, one of which was a novel mtDNA haplotype in 1997 and 2007 samples. In addition, there was no clear difference in the haplotype frequency through the time span. These results suggest that the low genetic variation of the island population persisted for the last hundred years. It is thus nearly impossible for the island population to recover its genetic variation in isolation. Conservation plans for this species should therefore include the promotion of genetic exchanges between the continental and island populations, such as through artificial introduction to Hokkaido.

Changes of mercury contamination in red-crowned cranes, Grus japonensis, in East Hokkaido, Japan.

Red-crowned cranes (Grus japonensis) are native to eastern Hokkaido (island population), in contrast to the mainland, which migrates between the Amur River basin and eastern China-Korea peninsula. During the 1990s we found that Red-crowned cranes in Hokkaido were highly contaminated with mercury: however, the source was unknown. We investigated the time trend of mercury contamination in Red-crowned cranes. Total mercury levels in the livers and kidneys from cranes dead in the 2000s were lower than those dead in the 1990s. Feather is a major pathway of mercury excretion for many bird species and is used as an indicator of blood mercury level during feather growth. As internal organs from the specimens collected before 1988 were not available, we analyzed the flight feather shavings from stuffed Red-crowned cranes dead in 1959-1987 and found that the mercury level of feathers from cranes dead in the 1960s and 1970s was not more than those from the cranes dead in the 2000s. These results suggest that mercury contamination in Red-crowned cranes in Hokkaido decreased temporally during the 1990s-2000s. This indicates the possible occurrence of some mercury pollution in Red-crowned cranes' habitat in this region in the 1990s or before.

Accurate analysis of prevalence of coccidiosis in individually identified wild cranes in inhabiting and migrating populations in Japan.

Eimeria gruis and E. reichenowi cause coccidiosis, a major parasitic disease of cranes. By non-invasive molecular approaches, we investigated the prevalence and genetic characterization of pathogens in two Japanese crane habitats; one is Hokkaido inhabited by the endangered red-crowned crane, and the other is Izumi in Kyushu where populations that consist mainly of vulnerable hooded and white-naped cranes migrate in winter. The non-invasively collected faecal samples from each wintering population were first subjected to host genomic DNA-targeted analyses to determine the sample origin and avoid sample redundancy. Extremely high prevalence was observed in the Izumi populations (> 90%) compared with the Hokkaido population (18-30%) by examining 470 specimens by microscopy and PCR-based capillary electrophoresis (PCR-CE), using genetic markers in the second internal transcribed spacer (ITS2). Correspondence analysis of PCR-CE data revealed differences in community composition of coccidia between hooded and white-naped cranes. 18S rRNA and ITS2 sequences were determined from single oocysts excreted by red-crowned and hooded cranes. Phylogenetic analysis of 18S rRNA suggested that E. reichenowi was polyphyletic while E. gruis was monophyletic. Together with PCR-CE data, these results indicate different host specificity among the E. reichenowi type. Our data suggest that E. reichenowi comprises multiple species.

Hepatic transcriptional profile and tissue distribution of cytochrome P450 1-3 genes in the red-crowned crane Grus japonensis.

The endangered red-crowned crane (Grus japonensis) is a protected species in eastern Hokkaido and injured specimens are treated with medication. The present study aimed at understanding the expression profiles of cytochrome P450 (CYP) 1-3 genes in red-crowned crane tissues. We used 14 individuals found dead in the wild in eastern Hokkaido or in Kushiro City Zoo. Nine CYP1-3 genes expressed in the liver of the red-crowned crane were identified by high-throughput sequencing, and phylogenetically classified as CYP1A5, CYP2C23, CYP2C45, CYP2D49, CYP2G19, CYP2U1, CYP2AC1, CYP3A37, and CYP3A80. Based on the quantitative real-time PCR of 13 samples, the rank order of their median expression levels was as follows: CYP3A37 > CYP2AC1 > CYP2C45 > CYP2D49 > CYP2G19 > CYP1A5 > CYP3A80 > CYP2C23. The tissue distribution of the CYP transcripts indicated that many of the CYP1-3 genes examined were mainly expressed in the tissues where drug metabolism occurs, such as the liver, kidneys, and lungs. We found that CYP3A37 was dominant at the transcript level in the liver, indicating it might play a crucial role in liver physiology and xenobiotic metabolism. Similarly, an "orphan" CYP2AC1 was expressed at relatively high levels in the kidneys and liver, suggesting a possible role in renal and liver physiology and xenobiotic metabolism. Our results establish a foundation for future studies on red-crowned cranes aiming to further understand drug sensitivity and develop medication protocols, but also contribute to national and local projects for the conservation of red-crowned crane.

Laparoscopic resection of schwannoma of the ascending colon.

Schwannomas of the colon are rare and difficult to diagnose preoperatively. We report a case of schwannoma of the ascending colon that was resected laparoscopically. A 64-year-old woman was referred to our hospital by her local clinic for further evaluation and management of a submucosal tumor of the ascending colon. A definitive preoperative diagnosis could not be reached despite examinations. Gastrointestinal stromal tumor, leiomyoma and lymphoma were the differential diagnoses. We performed a laparoscopic right hemicolectomy with D2 lymph node dissection. Histological findings with hematoxylin-eosin staining revealed spindle-like tumor cells, and immunohistochemical analysis showed that the tumor was positive for S-100 but negative for c-kit, CD34, smooth muscle actin and desmin, with a Ki-67 index of <5%. Thus, the diagnosis in this case was benign schwannoma of the ascending colon.

Report of a case: Retroperitoneal mucinous cystadenocarcinoma with rapid progression.

INTRODUCTION: Retroperitoneal mucinous cystic neoplasms are uncommon, and little is known about the etiology of the disease. Malignant forms of these are extremely rare. Here, we report a case of primary retroperitoneal mucinous cystadenocarcinoma (PRMC), which demonstrated unexpectedly aggressive progression despite finding only a limited area of adenocarcinoma. PRESENTATION OF CASE: A 62-year-old woman with a complaint of abdominal discomfort was admitted to the hospital. Abdominal CT and MRI showed multiple large retroperitoneal cysts dislocating the right kidney nearly to the center of the abdomen. Transabdominal resection of the cysts was performed. Those cysts contained 1100ml of mucinous fluids in total. Cytological examination of those fluids revealed no malignant cells. The cyst wall was lined with mucinous epithelial cells, and contained some ovarian-type stroma. Also, there was a focal area of adenocarcinoma in the cyst wall, and the lesion was diagnosed as primary retroperitoneal mucinous cystadenocarcinoma. Eight months later, the patient developed lumbar bone metastasis. Chemotherapy with S-1, an oral fluoropyrimidine, and docetaxel had been begun immediately; however, the disease had rapidly spread in the retroperitoneum. Eventually, the patient died of the disease 15 months after surgery. DISCUSSION: Retroperitoneal mucinous cystic neoplasms are considered to be metaplasia of embryonal coelomic epithelium. Complete excision without rupture is essential. However, variance of biological aggressiveness might exist in PRMCs. CONCLUSION: Retroperitoneal mucinous cystadenocarcinoma is a rare tumor, and it is urgently necessary to elucidate the etiology of an effective therapy for the disease.

Incidence of Helicobacter Pylori Infections and Therapy for Anastomotic ulcers and Residual Gastritis in Patients with Residual Stomachs.

Seventy-seven patients with the vestibular portion of their stomachs resected (residual stomach) due to ulcers or stomach cancers were included in this study. Fifty-five (71.4%) of these patients had Helicobacter pylori infections. H. pylori was found in all patients with stomach cancer, esophageal cancer, or esophageal venous varicosity, and in 86.7% of patients with esophagitis, 78.6% with residual gastritis, 70.0% with anastomotic ulcers, and in 51.6% of patients with normal mucosa in their resected stomachs. The in vitro antibacterial activities of amoxicillin, erythromycin, clarithromycin, minocycline, nalidixic acid, ofloxacin, lansoprazole, and omeprazole were determined against 10 clinical isolates of H. pylori. All agents except ofloxacin, nalidixic acid and omeprazole showed satisfactory bactericidal activities. The clinical and bacteriological efficacies of a combined treatment regimen with lansoprazole and clarithromycin were evaluated in 10 patients with anastomotic ulcers and 6 patients with residual stomach inflammation after informed consent. The clinical efficacy of this treatment in these 16 patients was excellent for 3 patients, good for 12 patients, and fair for the other patient. Collectively, the treatment was effective in 15/16 (93.8%) of patients. The bacteriological efficacy of this treatment regimen was evaluated in 14 patients, with no evidence of H. pylori 13 (92.9%) patients.

Clinical trial of in-situ hybridization method for the rapid diagnosis of sepsis.

We evaluated the utility of in-situ hybridization (ISH) for the rapid diagnosis of sepsis. We applied this approach to polymorphonuclear neutrophil (PMN)-rich smears from patients with suspected bacterial infection. Positive results by ISH were obtained in the smears of 123 of 292 patients (42%), while only 32 of the 292 (11%) were positive by blood culture. These findings indicate that ISH is almost four times more sensitive than the culture method for the detection of sepsis. ISH results are obtained within 1 day, while 1 day to 2 weeks is required for the results of blood culture. Blood culture and ISH methods detected the same bacteria in two patients. ISH also successfully identified the same bacteria in blood and PMN-rich body fluid (bronchoalveolar lavage samples) in 6 patients. In 19 patients, ISH of blood detected the same bacteria as those found in subcultures from other sources (e.g., stool, sputum, nasal cavity). We discuss these results in comparison with blood culture results in terms of evaluating a rapid approach to the management of patients with sepsis.

Gastric cancer with multiple intramural metastases and metastasis to the small intestine which eventually developed Trousseau's syndrome: report of a case.

Here we report a rare case of Trousseau's syndrome in a patient with gastric cancer with multiple intramural metastases and metastasis to the small intestine. A 70 year-old male complaining of appetite loss and weight loss of 7 kg within 3 months was admitted to hospital. Esophagogastroduodenal endoscopy revealed an advanced gastric cancer at the pylorus almost occluding the outlet of the stomach, and multiple ulcerative lesions throughout the stomach. A biopsy showed poorly differentiated adenocarcinoma. The patient underwent total gastrectomy. During surgery, part of the distal ileum was found to be abnormally firm and approximately 1 m of the ileum with the cecum colon was resected. Pathologic examination confirmed poorly differentiated adenocarcinoma at the pylorus and multiple intramural metastases in most other areas of the stomach. Lymph node metastases were confirmed in 12 out of 40 harvested regional lymph nodes including one positive paraaortic lymph node. The resected ileum contained multiple tumors with ulceration. Massive lymphatic invasion in the stomach and the small intestine was observed, which strongly suggested lymphatic spread of the gastric cancer. The patient was discharged on post-operative day 21; however, 2 months after surgery, he developed multiple cerebral thromboembolisms and died 2 weeks later.

Origin of three Red-crowned Cranes Grus japonensis found in northeast Honshu and west Hokkaido, Japan, from 2008 to 2012.

The Red-crowned Crane Grus japonensis is an endangered species that has two separate breeding populations, one in the Amur River basin and the other in north and east Hokkaido, Japan. So far, only two (Gj1 and Gj2) and seven (Gj3-Gj9) haplotypes in D-loop of mtDNA were identified in Japan and in the continent, respectively. We obtained feathers from three cranes found in northeast Honshu. The crane in Akita in 2008, which also arrived at west Hokkaido, had a novel haplotype (Gj10). Another crane in Akita in 2009 showed a heteroplasmy (Gj7 and a novel type, Gj12). The third crane in Miyagi in 2010 also showed another type, Gj11. These results suggest that three Red-crowned Cranes appeared in Honshu and west Hokkaido were from the continent.

Metastatic Small Bowel Tumor from Descending Colon Cancer with Extensive Hematogenous or Lymphogenous Spread: Survey of the Japanese Literature.

We present the case of a 68-year-old female patient who was diagnosed with cancer of the descending colon in July 1994 and underwent partial resection of the colon (type 2, moderately to well differentiated adenocarcinoma, se, ly1, v1, n(-)). In April 1996, she was admitted to a nearby hospital for symptoms of ileus, which improved at the hospital. However, she was referred to our hospital for melena. In blood test, Hb was 8.7 g/dl, showing anemia, and carcinoembryonic antigen level was elevated to 50.7 ng/ml. Abdominal CT and small bowel series showed only mild expansion of the small bowel, suggesting no obvious occlusion. Abdominal surgery was performed in May 1995 for repeated development of ileus symptoms and suspicion of bleeding from the small bowel. Since the findings of the abdominal surgery showed a circular tumor in the lower ileum, partial resection of the small bowel was performed. Histopathological examination showed type 3, moderately to well differentiated adnocarcinoma, se, ly2, v0, n = 1/13. The principal tumor was located within the subserosa and grew up exclusively through the muscularis propria and the submucosa, into the mucous layer. The mucosa remained slightly on the surface layer. Based on these findings, the patient was diagnosed with metastasis of descending colon cancer to the small bowel. Her prognosis was good, and neither metastasis nor redevelopment of the cancer have been confirmed to date, 11 years and 7 months since the surgery.

Genetical identification of coccidia in red-crowned crane, Grus japonensis.

We genetically analyzed eimerian oocysts isolated from the red-crowned crane (Grus japonensis) in Hokkaido, a northern island of Japan. Two types of oocysts of which shapes were similar to Eimeria gruis and E. reichenowi were found. Nearly the total length of the 18S ribosomal RNA gene (about 1.7 kbp in length) was amplified from single oocyst of each type and was sequenced. The respective sequences showed high similarity to those of published partial sequences (349 bp) of E. gruis type oocyst and E. reichenowi type oocyst isolated from the hooded crane (G. monacha) and the white-naped crane (G. vipio) in Izumi, in the southern island of Japan. Phylogenetic analysis indicated that E. gruis type and E. reichenowi type are different species, and suggested that these crane coccidia have evolved independently from the intestinal parasitizing Eimeria species.

Study of nasopharyngeal bacterial flora. Variations in nasopharyngeal bacterial flora in schoolchildren and adults when administered antimicrobial agents.

Changes in nasopharyngeal bacterial flora in adults with acute upper respiratory tract infection on administration of antimicrobial agents were investigated, and how these changes contrasted with those in children. Many patients with acute sinusitis due to allergies, and patients with malignancy and diabetes mellitus were included in the investigation. The detection rates of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis, the major bacteria of acute otitis media (AOM), were 22%, 10%, and 7% respectively, which were significantly lower than those for children. Gram stain examination of nasopharyngeal swab samples showed a significant relation between leukocyte infiltration and the detection amount of S. pneumoniae (P = 0.0086). A significant relation (P = 0.0134) was also observed when H. influenzae was simultaneously detected. No significant change in the three major AOM bacteria present in nasopharyngeal bacterial flora after administration of antimicrobial agents was observed. However, all S. pneumoniae and H. influenzae detected after antimicrobial agent administration had the beta-lactam-resistance gene. It was observed that a significant improvement in leukocyte infiltration occurred 6 to 10 days after antimicrobial agent administration. In contrast, a significant improvement in children was observed at 2 to 5 days. In the adult subjects, this improvement was probably due to spontaneous remission rather than the effect of the antimicrobial agents. Although investigation of the long-term administration of antimicrobial agents was also conducted, its benefits for the patients were not elucidated.

Study of nasopharyngeal bacterial flora. Second report. Variations in nasopharyngeal bacterial flora in children aged 6 years or younger when administered antimicrobial agents. Part 1.

In our first report, we investigated nasopharyngeal bacterial flora related to penicillin-resistant Streptococcus pneumoniae (PRSP) and beta-lactamase-negative ampicillin-resistant Haemophilus influenzae (BLNAR) and their relation to acute upper respiratory tract infection (AURTI). This report analyzes the results of a study of nasopharyngeal bacterial flora before the administration of antimicrobial agents in 172 AURTI patients aged 6 years or younger. In addition to Gram staining, microscopic observation, and culturing, a polymerase chain reaction (PCR) method was used to identify PRSP (gPRSP) and BLNAR (gBLNAR) drug-resistant genes. Of the patients analyzed, 90% had acute otitis media (AOM) and were aged 2 years or younger. The antimicrobial agents administered were amoxicillin (34%), clavulanic acid/amoxicillin (11%), cefditren pivoxil (CDTR-PI) (43%), and others (12%). This was particularly true for patients administered CDTR-PI, among whom there were many who had already suffered one or more episodes of AOM by the age of 1 year or younger, and many in which gPRSP were detected (P < 0.01). There was a significant relation between the degree of nasopharyngeal inflammation indicated by leukocyte infiltration images and the amount of S. pneumoniae and H. influenzae detected, which are the main pathogenic bacteria causing AOM (P < 0.01). In addition to leukocyte infiltration images, there were cases in which shedding of ciliated cells was observed and/or giant monocytic cells. Both nasopharyngeal leukocyte infiltration images and/or shed cell findings observed in infant AURTI cases are important indices for the prompt detection of gPRSP and/or gBLNAR and appropriate doses of antimicrobial agents.

Study of nasopharyngeal bacterial flora. Second report. Variations in nasopharyngeal bacterial flora in children aged 6 years or younger when administered antimicrobial agents. Part 2.

This report focuses on changes in the nasopharyngeal bacterial flora before and after administration of antimicrobial agents in 172 cases of acute upper respiratory infection in patients aged 6 years or younger. The antimicrobial agents administered were amoxicillin (AMPC) (34%), clavulanic acid/amoxicillin compound (11%), cefditren pivoxil (CDTR-PI) (43%), and others (12%). Changes in nasopharyngeal bacterial flora were investigated with reexaminations conducted after 2-5 days (day 2-5 subgroup), 6-10 days (day 6-10 subgroup), and 11 days and thereafter. There was a significant reduction in the Streptococcus pneumoniae detected in the group administered AMPC (AMPC group) in the day 2-5 subgroup and the day 6-10 subgroup. There was also a significant decrease in H. influenzae in the group administered CDTR-PI (CDTR-PI group) in the day 2-5 subgroup. From this it was inferred that for the most part significant changes in infectious nasopharyngeal bacteria occurred in the day 2-5 subgroups. However, a significant improvement in the degree of inflammation, as indicated by leukocyte infiltration images for the AMPC group, was observed in the day 2-5 subgroup, and for the CDTR-PI group in the day 6-10 subgroup. On the other hand, in both the antimicrobial agent groups, S. pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis were newly detected at reexamination. Furthermore, a difference in the incidence of these bacteria was observed between the 2 antimicrobial agent groups. It was suggested that such phenomena related to the survival of resistant strains or a recurrence otitis media.

Study of upper respiratory tract bacterial flora: first report. Variations in upper respiratory tract bacterial flora in patients with acute upper respiratory tract infection and healthy subjects and variations by subject age.

With the appearance of penicillin-resistant Streptococcus pneumoniae, there has been increasing debate concerning antimicrobial treatments for acute upper respiratory tract infection (AURTI) and acute otitis media in children. This study compares the nasopharyngeal bacterial flora in patients with AURTI (AURTI group; 710 subjects) and healthy subjects (HS group; 380 subjects). The comparisons were made between subjects aged 6 years or younger (0-6 subgroup: 330 subjects), between 7 and 74 years (7-74 subgroup: 668 subjects), and 75 years and older (92 subjects), because the subjects were subgrouped as described above dependent on the maturity of the protective immunity. In the HS group 7-74 subgroup, viridans group streptococci, Staphylococcus aureus, coagulase-negative staphylococci, and Corynebacterium sp. with a detection rate of 10% or more were classified as normal nasal flora (NNF), and Streptococcus pyogenes, Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis were classified as drum cavity pathogens (DCP). In the 0-6 subgroup, although the detection rate for DCP bacteria in the AURTI group tended to be high, it did not reach a significant difference, whereas the detection rate for NNF bacteria was significantly lower. This trend was also observed to some degree in the other age subgroup. In the 0-6 subgroup, leukocyte infiltration observed with a microscope indicated the closest relationship between S. pneumoniae detection rate and detection quantity. These results suggest that in the 0-6 subgroup the tendency for patients with AURTI to have NNF bacteria as well as DCP bacteria should be taken into consideration.