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The name Paragonimus westermani (Kerbert, 1878) is commonly applied to members of a species complex that includes the well-known Asian lung fluke of medical and veterinary importance. Unambiguous molecular and morphological evidence showing the presence of a member of the complex in India has recently been published. In the present study we report the occurrence of 2 more members of the P. westermani complex in northeastern (NE) India. Surveys of the freshwater crabs Maydelliatelphusa lugubris in NE India revealed 2 morphologically distinct types of lung fluke metacercariae. Phylogenetic analyses, using DNA sequences from ITS2, 28S and cox1 gene regions indicate that these lung metacercariae belong to P. westermani complex. Type 1 metacercariae have a more basal position within the complex whereas type 2 metacercariae are closely related to the relatively derived forms of P. westermani from NE Asia (Japan, Korea, China) and Vietnam. A third type of metacercaria (type 3), detected in another crab host, Sartoriana spinigera in Assam, was phylogenetically close to P. siamensis, also a member of the P. westermani group. Molecular evidence has demonstrated the existence of 3 genotypes of lung flukes within the Paragonimus westermani complex in NE India. Two of these were previously unknown.

Purpose: This phase I/II study was designed to evaluate a combination of irinotecan and S-1 a new regimen for salvage chemotherapy in patients with advanced or metastatic non-small cell lung cancer (NSCLC)., Methods: The study group comprised patients with advanced or metastatic NSCLC who had previously received at least one platinum-containing chemotherapy. Patients received irinotecan on days 1, 15 and oral S-1 (40 mg/m(2) twice daily as a fixed dose) on day 1 to 14 of a 28-day cycle., Results: In the phase I part, irinotecan was given in escalating doses of 70 (Level 1), 80 (Level 2), and 90 mg/m(2) (Level 3). Three of the 5 patients given Level 3 had dose-limiting toxicity, and Level 2 (80 mg/m(2) of irinotecan) was designated as the recommended dose. In phase II, 38 patients received a median of 7.4 cycles of irinotecan at the recommended dose. The overall response rate was 15.8 % (90 % confidence interval (CI): 6.1-25.5 %), and the median progression-free and overall survival times were 4.5 months (95 % CI: 3.5-5.0) and 15.0 months (95 % CI: 9.5-20.6) months, respectively. Toxicity was generally mild. Grade 3 or higher toxicity included neutropenia in 17.9 % of the patients, thrombocytopenia in 5.1 % and nausea in 7.7 %., Conclusion: Combination chemotherapy with S-1 and irinotecan was considered an effective salvage regimen in patients with advanced or metastatic NSCLC.

Arginine kinase (AK) is a member of the phosphagen kinase family. AK plays a major role in cellular energy metabolism in invertebrates including nematodes. In the present study, we performed the direct immunofluorescence test to determine the immunolocalization of AK in different stages of the life cycle (eggs, larvae, and adult worms) of Toxocara canis, Toxocara vitulorum, and Ascaris lumbricoides. Our results indicated variable levels of expression of AK in different stages. Moreover, strong fluorescence was observed in cleaving eggs than in dormant eggs. The highest activity of the enzyme was observed in the fully developed eggs. This may be due to high expression of AK in embryonic development, which is associated with increased energy demand due to cleavage and cellular differentiation. Surprisingly, expression of AK is significantly higher in the middle part and posterior end compared to anterior end of the larvae. In addition, AK is highly concentrated in cellular and metabolically active parts of the body such as hypodermis, muscle, intestine, ovaries, oviducts, and uterus, while it is absent in noncellular areas like cuticle. The present study revealed the presence of AK in T. canis, A. lumbricoides, and T. vitulorum and that it plays a major role in energy metabolism of these nematodes. Interestingly, antiserum was prepared against the recombinant T. canis AK and reacts with the native AKs of T. canis, A. lumbricoides, and T. vitulorum. AK levels could vary in relation to maximum potential rates of ATP turnover, oxidative capacity, and energy output. Further studies on subcellular localization of AK in these important helminths provide new information for researchers to develop effective anthelmintics against the parasites of veterinary and of public health importance.

Mas-related G-protein coupled receptor member D (MRGPRD) is a G protein-coupled receptor (GPCR) which belongs to the Mas-related GPCRs expressed in the dorsal root ganglia (DRG). In this study, we investigated two novel ligands in addition to beta-alanine: (1) beta-aminoisobutyric acid, a physiologically active substance, with which possible relation to tumors has been seen together with beta-alanine; (2) diethylstilbestrol, a synthetic estrogen hormone. In addition to the novel ligands, we found that transfection of MRGPRD leads fibroblast cells to form spheroids, which would be related to oncogenicity. To understand the MRGPRD novel character, oncogenicity, a large chemical library was screened in order to obtain MRGPRD antagonists to utilize in exploring the character. The antagonist in turn inhibited the spheroid proliferation that is dependent on MRGPRD signaling as well as MRGPRD signals activated by beta-alanine. The antagonist, a small-molecule compound we found in this study, is a potential anticancer agent.

To elucidate the function of MAS-related GPCR, member D (MRGD) in cancers, we investigated the in vitro and in vivo oncogenic function of MRGD using murine fibroblast cell line NIH3T3 in which MRGD is stably expressed. The expression pattern of MRGD in clinical samples was also analyzed. We found that overexpression of MRGD in NIH3T3 induced focus formation and multi-cellular spheroid formation, and promoted tumors in nude mice. In other words, overexpression of MRGD in NIH3T3 induced the loss of contact inhibition, anchorage-independent growth and in vivo tumorigenesis. Furthermore, it was found that the ligand of MRGD, beta-alanine, enhanced spheroid formation in MRGD-expressing NIH3T3 cells. From investigation of clinical cancer tissues, we found high expression of MRGD in several lung cancers by immunohistochemistry as well as real time PCR. Based on these results, MRGD could be involved in tumorigenesis and could also be a novel anticancer drug target.

Background: Chronic inflammation, imbalance of proteolytic and antiproteolytic activities, oxidative stress, and apoptosis of lung structural cells contribute to the pathogenesis of COPD. There is increasing evidence that carboxymethylcysteine (also known as carbocisteine) (CMC), which is commonly used for its mucoactive property, has diverse pharmacologic actions, including significant antioxidant activity. We hypothesize that CMC protects against cigarette smoke extract (CSE)-induced emphysema in rats via its antioxidant action., Methods: Sprague-Dawley rats were divided into four groups (n = 6 in each group): control group, CSE group, CSE + 125 mg/kg/d of CMC group, and CSE + 250 mg/kg/d of CMC group. The CSE was injected intraperitoneally once a week for 3 weeks, and CMC was administered daily via a gastric gavage for the same duration. Antioxidant activity in the pulmonary and serum levels, apoptotic index, caspase-3 activity, and matrix metalloproteinase (MMP)-2 and MMP-9 activities in lung tissues were measured., Results: CMC significantly protected against alveolar enlargement and parenchymal destruction in rats injected with CSE, resulting in prevention of the development of CSE-induced emphysema in the rats. CMC significantly protected the antioxidant activity in both the pulmonary and systemic levels, reduced pathologic apoptosis, and inhibited MMP-2 and MMP-9 activities in the lungs of rats with CSE-induced emphysema., Conclusions: CMC protected against the development of CSE-induced emphysema in rats. The molecular mechanisms that were involved with stabilizing the biologic antioxidant activity resulted from the administration of CMC, which was connected to the inhibition of apoptosis and the reversal of the imbalance of proteolytic and antiproteolytic enzyme activities, eventually achieving the protection of the alveolar architecture of rats with emphysema.

Avian influenza virus vaccines produced in oil-emulsified inactivated form with antigen content of at least 160 hemagglutinin units (HAU) induced immunity in birds. However, in addition to enhancing the effect of the adjuvant(s), other additional supplemented biological compounds included in inactivated vaccines could produce higher levels of antibody. We examined in chickens, Vietnamese ducks, and muscovy ducks the adjuvant effect of Sophy β-glucan (SBG), a β-1,3-1,6 glucan produced by the black yeast Aureobasidium pollulans strain AF0-202, when administered with an avian influenza H5 subtype vaccine. In Experiment 1, 40 chickens (ISA Brown hybrid), allocated to four groups of ten each, were immunized with Oil-H5N1(VN), Oil-H5N1(CN), Oil-H5N2(CN), and saline (control group), respectively. In Experiment 2, chickens (ISA Brown hybrid), muscovy ducks (French hybrid), and Vietnamese ducks (indigenous Vietnamese) were used to further assess the effect of SBG on immunogenicity of the Oil-H5N1(VN) Vietnamese vaccine. ELISA and hemagglutination inhibition (HI) assays were used to assess the antibody response. The H5 subtype vaccines initiated significantly higher immune responses in the animals dosed with SBG, with 1.0-1.5 log2 higher HI titers and 10-20% ELISA seroconversion, compared with those not dosed with β-glucan. Notably, some of the animals dosed with SBG induced HI titers higher than 9.0 log2 following boosting immunization. Taken together, our serial studies indicated that SBG is a potential effector, such as enhancing the immune response to the H5 vaccines tested.

The optimal chemotherapeutic regimen for inoperable thymic carcinoma remains uncertain and little information is available regarding the usefulness of salvage chemotherapy. S-1, a newly developed oral fluorouracil antitumor drug, has been reported to be effective in the treatment of gastrointestinal tumors and non-small cell lung cancer. This case study reports a case of chemorefractory thymic cancer with a good response to S-1 monotherapy. S-1 was used as sixth-line chemotherapy and the response was the first remarkable tumor regression in the patient's clinical course. S-1 appears to have significant activity against thymic carcinoma.

A 66-year-old man was diagnosed with autoimmune pancreatitis in February 2009 and started 40 mg of oral prednisolone followed by a maintenance dose of 5 mg daily. The patient developed a cough in October 2010 and visited our division. He had a high serum concentration of immunoglobulin (Ig) G4 and his chest computed tomography showed airway stenosis without bilateral hilar lymphadenopathy (BHL). The bronchial biopsy specimens revealed lymphoplasmacytic infiltrations with IgG4-positive/IgG-positive plasma cells of more than 50%. Thus, we diagnosed the airway lesion with IgG4-related airway involvement. This is the first report of a patient with IgG4-related airway involvement without BHL.

A 43-year-old-man was diagnosed as having primary mediastinal nonseminomatous germ cell tumor based on percutaneous biopsy and an elevated serum concentration of alpha-fetoprotein (AFP). Although AFP decreased after chemotherapy, the mass size grew and developed tracheal compression by the tumor. The patient was treated with mechanical ventilation and subsequent stent implantation for keeping airway patency. After three cycles of chemotherapy and normalization of AFP, the increased mass was successfully resected and the pathological examination demonstrated a mature teratoma. This case showed a rare clinical manifestation of mediastinal growing teratoma syndrome and successful outcome by multimodal therapies.

Most mantle cell lymphoma patients show remarkable disseminated disease at the initial diagnosis. We describe two cases of mantle cell lymphoma mainly involving thoracic lesions at the initial presentation of the disease. The clinical presentations were right hilar lymphadenopathy in one case and right pleural thickness in the other. The diagnosis of mantle cell lymphoma was confirmed by immunohistochemistry, including CD5, CD20, and cyclin D1, and the presence of t(11 ; 14)(q13 ; q32) by fluorescence in situ hybridization. These thoracic manifestations at the initial diagnosis should be taken into consideration for the clinical spectrum of mantle cell lymphoma.

This study was carried out to determine the effect of Sophy β-glucan on immunity and growth performance in broilers. One group was treated with 1% β-glucan ad libitum with water and the other group was kept as the control. Vaccination for Infectious Bursal Disease was carried out on days 16 and 21. Blood samples were collected from birds, and antibody titres against IBD were measured. The mean body weight of the β-glucan treated group was significantly (P<0.05) higher than that of the control group. The mean antibody titres measured on days 25, 36 and 42 were significantly (P<0.05) higher in 1% β-glucan treated group than that of the control group, suggesting the presence of immune stimulating effect of β-glucan.

The role of second-line and salvage chemotherapy in malignant pleural mesothelioma treatment is not yet established. We report a case of relapsed malignant pleural mesothelioma in which the patient failed to respond to pemetrexed-based chemotherapy but was successfully treated with gemcitabine and vinorelbine. The patient underwent a left extrapleural pneumonectomy. Three years later she developed anterior chest wall and retroperitoneal masses. Histological findings revealed metastases from the malignant pleural mesothelioma. Although two cycles of carboplatin plus pemetrexed chemotherapy were administered, she had progressive disease. Then, 1000 mg/m(2) gemcitabine and 25 mg/m(2) vinorelbine were administered every 2 weeks. The chemotherapy regimen was tolerated well, and the tumors were remarkably reduced. She was treated with 12 cycles of gemcitabine plus vinorelbine, and 8.5 months of progression-free survival was observed. Gemcitabine plus vinorelbine chemotherapy may be a candidate regimen for salvage chemotherapy against malignant pleural mesotheliomas.

Due to the possible emergence of resistance and safety concerns on certain treatments, development of new drugs against parasites is essential for the effective control and subsequent eradication of parasitic infections. Several drug targets have been identified which are either genes or proteins essential for the parasite survival and distinct from the hosts. These include the phosphagen kinases (PKs) which are enzymes that play a key role in maintenance of homeostasis in cells exhibiting high or variable rates of energy turnover by catalizing the reversible transfer of a phosphate between ATP and naturally occurring guanidine compounds. PKs have been identified in a number of important human and animal parasites and were also shown to be significant in survival and adaptation to stress conditions. The potential of parasite PKs as novel chemotherapeutic targets remains to be explored.

Evidence for the presence of lung flukes of the Paragonimus westermani in India remains scant. In particular, evidence based on morphology of adult worms is lacking. Metacercariae of the genus Paragonimus, recovered from crabs in two regions of northeastern India, were raised to adulthood in laboratory rats. Morphologically, these worms appear to be P. westermani. DNA sequences from the second internal transcribed spacer (ITS2) and a portion of the ribosomal large subunit gene (28S) of the nuclear ribosomal RNA gene repeat, as well as fragments of the mitochondrial cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) genes, all supported this identification. Molecular phylogenetic methods were used for studying the relatedness of these Indian flukes with counterparts from southeast and far-east Asia. Molecular data showed that Indian representatives of the P. westermani complex represent a distinct lineage. It is unclear whether the Indian form can cause disease in humans as some members of the complex do elsewhere., (Copyright 2010 Elsevier B.V. All rights reserved.)

In the present study, we determined the complete mitochondrial (mt) genome sequence (13,839bp) of parasitic nematode Setaria digitata and its structure and organization compared with Onchocerca volvulus, Dirofilaria immitis and Brugia malayi. The mt genome of S. digitata is slightly larger than the mt genomes of other filarial nematodes. S. digitata mt genome contains 36 genes (12 protein-coding genes, 22 transfer RNAs and 2 ribosomal RNAs) that are typically found in metazoans. This genome contains a high A+T (75.1%) content and low G+C content (24.9%). The mt gene order for S. digitata is the same as those for O. volvulus, D. immitis and B. malayi but it is distinctly different from other nematodes compared. The start codons inferred in the mt genome of S. digitata are TTT, ATT, TTG, ATG, GTT and ATA. Interestingly, the initiation codon TTT is unique to S. digitata mt genome and four protein-coding genes use this codon as a translation initiation codon. Five protein-coding genes use TAG as a stop codon whereas three genes use TAA and four genes use T as a termination codon. Out of 64 possible codons, only 57 are used for mitochondrial protein-coding genes of S. digitata. T-rich codons such as TTT (18.9%), GTT (7.9%), TTG (7.8%), TAT (7%), ATT (5.7%), TCT (4.8%) and TTA (4.1%) are used more frequently. This pattern of codon usage reflects the strong bias for T in the mt genome of S. digitata. In conclusion, the present investigation provides new molecular data for future studies of the comparative mitochondrial genomics and systematic of parasitic nematodes of socio-economic importance., (2010 Elsevier B.V. All rights reserved.)

Hepatozoon species infections were detected in 4 of 5 heart samples from leopard cats (Prionailurus bengalensis) in Korea examined by light microscopy and PCR. Microscopically, the schizonts of the Hepatozoon species were observed within the parasitophorous vacuoles of unidentified host cells that were located between myocytes. The schizonts were approximately 31 +/- 4 x 19 +/- 3 microm in size; 3 mature schizonts contained approximately 24, 25, and 35 merozoites. In analyses of the partial 18S rRNA gene sequence, the Hepatozoon species infecting the Korean leopard cats was closely related to Brazilian (99.2-99.8% identical) and Spanish (97.0-98.2% identical) isolates of Hepatozoon felis.

Background: Pulmonary pleomorphic carcinoma is a very rare pulmonary malignant tumor which has various clinical manifestations and a poor prognosis., Case: A 74-year-old man presented with fever and weight loss of more than 10kg during 2 months from April 200X. Chest CT showed a nodule in the right upper lobe, and supraclavicular and mediastinal lymphadenopathy. Positron emission tomography (PET) comfirmed 18Ffluorodeoxyglucose accumulation in the sites. A left supraclavicular node biopsy demonstrated pulmonary pleomorphic carcinoma. The tumors responded well to gemcitabine plus docetaxel combination chemotherapy. This symptoms disappeared and the response continued for 1 year after 6 cycles of chemotherapy., Conclusion: We report a case of pulmonary pleomorphic carcinoma presenting with fever, which showed a marked response to chemotherapy.

In 2009, a 35-year-old female with Down syndrome was admitted to our hospital because of severe pneumonia caused by an infection with the novel swine-origin influenza (A/H1N1pdm) virus (S-OIV). A chest X-ray on admission revealed bilateral infiltration shadows. Although mechanical ventilation was administered because of the development of ARDS, the hypoxemia continued to progressed. We observed evidence of alveolar hemorrhage on evaluation of the patient using bronchofiberscopy. The bacterial examination was negative. Despite intensive care, including respiratory management with high-frequency oscillatory ventilation (HFOV), the patient's hypoxemia and hypotension progressed. We concluded that a cytokine storm due to the influenza infection with SIRS caused shock status, resulting in septic shock. We subsequently treated the patient with direct hemoperfusion with polymyxin B-immobilized fiber (PMX-DHP). The hypoxemia improved immediately. She was free from mechanical ventilation and discharged from the hospital by the 17th day of her hospitalization. PMX-DHP seems to improve hypoxemia in patients with severe ARDS who cannot maintain sufficient respiratory control under mechanical ventilation. This case is the first report about severe and life-threatening ARDS due to the novel influenza, in which PMX-DHP showed beneficial effects., (Copyright © 2010 Wiley-Liss, Inc.)

The effects of direct hemoperfusion with polymyxin B-immobilized fiber (PMX-DHP) treatment for acute exacerbation of interstitial pneumonia have been reported. Here, we report 3 severe cases of drug-induced interstitial lung disease (DI-ILD) that were treated with PMX-DHP. Two DI-ILD cases were caused by methotrexate, and the third case was caused by the herbal medicine sanmotsu-ogon-to. The arterial oxygen tension/inspiratory oxygen fraction ratio improved during PMX-DHP treatment in all 3 patients. Finally, 2 patients survived and 1 died. The study findings indicate that PMX-DHP treatment is a viable option for the management of acute respiratory failure in patients with DI-ILD.

The Hepatozoon species in the myocardium of Japanese martens (Martes melampus melampus) was characterized by histological and molecular methods. Histologically, granulomatous nodules with Hepatozoon sp. merozoites/gametocytes, or schizonts, or both were observed in the hearts of Japanese martens. The most frequently observed forms were merozoites/gametocytes within phagocytes; each host cell included a zoite, which was not microscopically identifiable as a merozoite or gametocyte. Schizonts were oval in shape and 36.9 ± 5.7 x 28.9 ± 3.4 μm in size; each schizont had approximately 20-60 nuclei. The size of the merozoites could not be measured because no mature schizonts were observed. In the analyses of the partial 18S rRNA gene sequence, it was strongly suggested that the Hepatozoon sp. in Japanese marten and the Hepatozoon sp. in pine marten (Martes martes) in Scotland were the same species.

The beta-glucans derived from yeast cell walls have been reported for having many immunomodulatory activities in vivo and in vitro. In this study, Aureobasidium-derived soluble branched (1,3-1,6) beta-glucan (Sophy beta-glucan) was checked for natural killer (NK) activity and for the production of IFN-gamma and IL-4 in Leishmania amazonensis infection. The main experiment was performed with a group of female C57BL/6 and BALB/c mice, orally supplemented with 5% of Sophy beta-glucan and infected with promastogotes of L. amazonensis (1 x 10(7)) into the footpad. Increase in the footpad thickness with time was observed in BALB/c mice in spite of the oral Sophy beta-glucan supplement, but it was less in C57BL/6 mice. The difference in overall mean footpad thickness between 'infection only' versus 'infection + glucan' groups was statistically significant (P < 0.001). High NK activity in C57BL/6 than BALB/c mice was observed in 'glucan only' group compared to the control group and also in 'infection + glucan' group compared to 'infection only' group. The difference in the NK activity among these groups was significant (P < 0.05). The IFN-gamma level increased at weeks 7 and 8 post-infection in C57BL/6 mice and was significantly high in 'infection + glucan' group compared to the 'infection only' group (P < 0.05). IL-4 levels did not increase up to detectable levels throughout the study. The results led a conclusion that Sophy beta-glucan enhances NK activity and cellular immunity in L. amazonensis-infected mice.

HIV-1 Nef accelerates the progression to AIDS by binding with and activating a Src kinase Hck, but underlying molecular basis is not understood. We revealed that Nef disturbed N-glycosylation/trafficking of a cytokine receptor Fms in an Hck-dependent manner, a possible trigger to worsen uncontrolled immune system. Here, we provide direct evidence that dys-regulated activation of Hck pre-localized to the Golgi apparatus causes this Fms maturation arrest. A striking change in Hck induced by Nef other than activation was its skewed localization to the Golgi due to predominant Golgi-localization of Nef. Studies with different Nef alleles and their mutants showed a clear correlation among higher Nef-Hck affinity, stronger Hck activation, severe Golgi-localization of Hck and severe Fms maturation arrest. Studies with a newly discovered Nef-Hck binding blocker 2c more clearly showed that skewed Golgi-localization of active Hck was indeed the cause of Fms maturation arrest. 2c blocked Nef-induced skewed Golgi-localization of an active form of Hck (Hck-P2A) and Fms maturation arrest by Nef/Hck-P2A, but showed no inhibition on Hck-P2A kinase activity. Our finding establishes an intriguing link between the pathogenesis of Nef and a newly emerging concept that the Golgi-localized Src kinases regulate the Golgi function.

Background and Objective: The SD-101 is a non-restrictive, sheet-like medical device with an array of pressure sensors, to detect sleep-disordered breathing by sensing gravitational alterations in the body corresponding to respiratory movements. This study evaluated the accuracy of the SD-101 for screening sleep apnoea-hypopnoea syndrome (SAHS) by comparison with polysomnography., Methods: Nocturnal polysomnography and SD-101 monitoring were conducted simultaneously and compared in 201 patients with suspected SAHS (suspected SAHS group) and 165 male employees of a transport company (screening group)., Results: Polysomnography revealed an AHI of <5, 5 < or = AHI < 15, 15 < or = AHI < 30, 30 < or = AHI < 60 and AHI > or = 60 events/h in 39, 35, 38, 68 and 21 subjects in the suspected SAHS group and 103, 34, 12, 12 and four subjects in the screening group, respectively. Central SAHS and obstructive SAHS were subsequently diagnosed in 11 (5.5%) and 135 (67.2%) of subjects in the suspected SAHS group and five (3.0%) and 39 (23.6%) of subjects in the screening group, respectively. Significant correlations were apparent between AHI and the respiratory disturbance index (RDI) measured with the SD-101 in both the suspected SAHS group (r = 0.88) and screening group (r = 0.92). Receiver operating characteristic curve analysis revealed 89.5% sensitivity and 85.8% specificity in identifying SAHS, using an RDI of 14.0 events/h., Conclusions: These findings suggest that the SD-101 is a useful device for screening SAHS.

A 64-year-old man was admitted to our hospital because of fever and dyspnea with marked hypoxemia and diffuse ground-glass opacities in bilateral lung fields revealed by a chest CT scan. He had used etanercept therapy for his rheumatoid arthritis. His PaO2/FiO2 had decreased to 130.4 Torr. On bronchoalveolar lavage, lymphocytes were elevated to 54.4% and bacteria culture was negative. We diagnosed drug-induced pneumonitis caused by etanercept, clinically and started high dose corticosteroid therapy. Despite his severe hypoxemia, the corticosteroid therapy and use of non-invasive positive pressure ventilation improved his condition. Interstitial lung disease induced by etanercept is rare, and a severe case requiring mechanical ventilation has never been reported. Because of the critical condition it can cause, it is suggested that evaluation of interstitial pneumonia is crucial.

Taurocyamine kinase (TK) was previously reported to be restricted to certain marine annelids; however, the present study has proven otherwise. The lung fluke Paragonimus westermani has a contiguous two-domain TK with a mass of 80216 Da consisting of 713 amino acid residues sharing higher sequence identity with molluscan arginine kinase (AK). Both domains of P. westermani TK have significant activity for the substrate taurocyamine and exhibited synergism during substrate binding. Since TK plays a key role in energy metabolism and is not present in mammals, inhibitors against P. westermani TK could be effective novel chemotherapeutic agents and could be utilized for the development of specific diagnostic tools for the detection of paragonimiasis.

Partial mitochondrial (mt) genome sequence (10,486bp) from the parasitic nematode Toxocara vitulorum was determined and its organization and structure compared with those of T. cati, T. canis and T. malaysiensis. The obtained mt genome sequence of T. vitulorum contains 10 protein-coding genes (cytochrome c oxidase subunits 1-3, Nicotinamide adenine dinucleotide dehydrogenase subunits 1-5, ATP synthase subunit 6 and cytochrome b), 14 transfer RNA genes and the large ribosomal RNA gene (rrnL), non-coding regions. ORF encoding for ATPase subunit 8 is not found in this partial mtDNA sequence. Five translation initiation codons were inferred, ATT, ATG, GTG, GTT and TTG. Most of the genes used TAG or TAA as a stop codon and two genes ended with a T. The gene arrangement and composition of the T. vitulorum mt genome is very similar to that of other Toxocara species mitochondrial genomes sequenced thus far. All genes are transcribed in the same direction, as other Toxocara species. This genome has a high A+T content (67.5%) and low G+C content (32.5%). Phylogenetic reconstruction based on aligned nucleotide sequences of seven taxa provided strong support that Toxocara vitulorum is more closely related to T. malaysiensis than to T. canis and T. cati.

A 76-year-old man with squamous cell lung cancer underwent right lower lobectomy in November, 2005. He was diagnosed with pT2N0M0, stage I B, and tegafur/uracil (UFT) was administered. On July, 2007, right hilar lymphadenopathy was detected and considered to be a recurrence. UFT was discontinued, and gemcitabine (GEM) and docetaxel (DOC) combination chemotherapy was initiated on August 21. He began to complain of fatigue, palpitation and dizziness since the fifth day of the administration, and anemia (hemoglobin: Hb 8.7 g/dL) was detected on the fifteenth day. On the twenty-second day of the administration, he was admitted to our hospital because of aggravation of anemia (Hb 6.5 g/dL). His anemia was diagnosed as immune hemolytic anemia based on the laboratory findings including a positive Coombs' test. He showed improvement with prednisolone therapy. The anemia was considered to be drug-induced. This case was extremely rare, and there are no reports on immune hemolytic anemia related to GEM and/or DOC.

Toxocara canis and Toxocara vitulorum are two important parasites of dogs and buffaloes with public health concern. The objectives of the present study are to identify molecular markers to discriminate these closely related parasites and to determine their phylogenetic position and genetic diversity within the genus Toxocara. Thus, two mitochondrial genes (complete ATPase 6 and partial small subunit ribosomal RNA (12S rDNA)), two nuclear ribosomal genes (second internal transcribed spacer region (ITS-2)), and part of the large subunit 28S region were analyzed. Nucleotide sequence (597 bp) and predicted amino acid sequences of the complete ATPase 6 gene (199 amino acids) of both species (T. canis and T. vitulorum) are similar in size with the Toxocara cati and Toxocara malaysiensis. There was 88% nucleotide similarity between T. canis and T. vitulorum and many transversions present in the 12S gene. Analyses of the ITS-2 and 28S regions revealed that the 28S region was more conserved (95% nucleotide similarity between T. canis and T. vitulorum) than the ITS-2 region (85%). This study has provided useful molecular markers for the molecular epidemiological investigation of Toxocara species. Further, phylogenetic analyses of the ITS-2 and 28S genes have indicated that the members of the genus Toxocara form a distinct group with reference to their definitive hosts.

Paragonimiasis is a common parasitic zoonosis in Cameroon and neighbouring countries in Western Africa. Serum, sputum and faecal samples were collected in an endemic area of South West Province, Cameroon, after administration of a questionnaire to identify individuals with appropriate symptoms and histories. Microscopic examination revealed eggs in sputum from 16 people, but none in any faecal sample. These 16 were among the 25 and 26 people, respectively, positive by ELISA and by immunoblot using Paragonimus africanus crude antigens. Copro-DNA detection was attempted using 23 faecal samples (18 from sputum egg-negative and five from sputum egg-positive individuals). Copro-DNA was detected in four of the five sputum egg-positive individuals. These results strongly suggest that: (1) serology is much more sensitive than sputum examination for diagnosis of paragonimiasis; and (2) a copro-DNA test may be more sensitive than a microscopic search for eggs in faeces. Molecular sequence data from ITS2 and cox1 genes confirmed that adult worms experimentally raised in cats were P. africanus and that eggs from sputum or other worm products from human faeces also belonged to this species. Based on these results, 26 of 168 persons (15.5%) were diagnosed as suffering from paragonimiasis.

Freshwater snails (family Paludomidae, genus Paludomus) were collected from streams in Hedeniya and Peradeniya (the campus of Peradeniya University), Kandy district, Central Province, Sri Lanka, and found to harbor rediae and cercariae of a Paragonimus sp. These larvae were identified as Paragonimus westermani by using ITS2 DNA sequences. The infection rates of P. westermani in Paludomus sp. in Hedeniya and Peradeniya were 0.1% (one of 1014) and 0.2% (two of 1006), respectively. The snail has not been identified to species in the present study. This is the first report of the snail host of Paragonimus in Sri Lanka.

A 1.9 kb nucleotide sequence of part of the mitochondrial (mt) genome covering the cox1-trnT-rrnL-trnC-rrnS region, and the order of the remaining mitochondrial protein-coding genes for S. nasale of Sri Lankan origin, has been determined for analysis of the possible placement of this species in the genus Schistosoma. The gene order of this species is similar to that of the African and Indian Schistosoma species, but strikingly different from the East Asian species. Analysis of an alignment of the 1.9 kb sequence with available sequences from other schistosomes indicated affinities with S. spindale (found in Sri Lanka) and African species (in particular S. intercalatum and S. haematobium). Phylogenetic trees inferred from the alignment including 1 kb of RNA (transfer RNA and ribosomal RNA) sequence for 8 other Schistosoma spp. and Fasciola hepatica as an out-group revealed that S. nasale is placed proximally to S. spindale, S. intercalatum, S. haematobium and S. mansoni in the African sub-group while the East Asian species are more distant. S. incognitum lies basal to the combined African/Indian clade. The mtDNA analysis strongly supports the hypothesis that S. nasale is closely affiliated with the African/Indian schistosome group rather than the East Asian Schistosoma species.

We report 8420 bp of DNA sequence data from the maxicircle (mitochondrial) genome of Leishmania major (MHOM/SU/73/5ASKH), a much larger portion of this genome than has been reported previously from any Leishmania species infecting humans. This region contains 10 partial and complete genes: 5 protein-encoding genes (COII, COIII, ND1, ND7 and Cyt b); two ribosomal RNA subunits (12S and 9S) and three unidentified open reading frames (MURF1, MURF4 (ATPase6) and MURF5), as in the lizard-infecting species L. tarentolae. The genes from L. major exhibit 85-87% identity with those of L. tarentolae at the nucleotide level and 71-94% identity at the amino acid level. Most differences between sequences from the two species are transversions. The gene order and arrangement within the maxicircle of L. major are similar to those in L. tarentolae, but base composition and codon usage differ between the species. Codons assigned for initiation for protein-coding genes available for comparison are similar in five genes in the two species. Pre-editing was identified in some of the protein-coding genes. Short intergenic non-coding regions are also present in L. major as they are in L. tarentolae. Intergenic regions between 9S rRNA and MURF5, MURF1 and ND1 genes are G+C rich and considered to be extensive RNA editing regions. The RNA editing process is likely to be conserved in similar pattern in L. major as in L. tarentolae.

Toxocariasis is a worldwide zoonotic disease caused by the ascarid nematode Toxocara canis. The most common method available for serodiagnosis of toxocariasis is an enzyme-linked immunosorbent assay (ELISA) test using Toxocara excretory-secretory antigen (TES). The present study describes the development of IgG-ELISA based on antiserum prepared against the recombinant arginine kinase of Toxocara canis. Antiserum was prepared against the purified recombinant arginine kinase (AK) using 6-week-old female Japanese white rabbits. Serum samples were collected from experimentally infected BALB/c and C57BL/6 mice at different time periods. The IgG-ELISA was performed using serum samples from mice (infected/uninfected) and TES antigen with antiserum prepared against the recombinant-AK. The optical density (OD450) was measured at 450 nm using a micro-plate ELISA reader. There were significant differences (P<0.01) in the absorbance between infected and control serum samples. Further, we obtained 100% sensitivity for the serum samples from T. canis-infected mice. Therefore, it is suggested that the recombinant-AK based IgG-ELISA could be applied for immunodiagnosis of human toxocariasis. However, it is necessary to evaluate the specificity of this recombinant antigen with similar geohelminth infections.

Morphological and genetic features of a new Hepatozoon species, Hepatozoon ursi n. sp., in Japanese black bear (Ursus thibetanus japonicus) were studied. Schizogonic developmental stages were observed in the lungs of Japanese black bears. The schizonts were sub-spherical in shape and 45.7+/-4.6 x 42.7+/-4.5 microm in size. Each mature schizont contained approximately 80-130 merozoites and 0-5 residual bodies. The merozoites were 7.0+/-0.7 x 1.8+/-0.3 microm in size. Intraleukocytic gametocytes were slightly curved, cigar-like in shape and had a beak-like protrusion at one end. The size of the gametocytes was 10.9+/-0.3 x 3.3+/-0.2 microm. The analyses of the18S rRNA gene sequences supported the hypothesis that H. ursi n. sp. is different from other Hepatozoon species. Mature Hepatozoon oocysts were detected in two species of ticks (Haemaphysalis japonica and Haemaphysalis flava) collected on the bears infected with H. ursi n. sp. Two measured oocysts were 263.2 x 234.0 microm and 331.8 x 231.7 microm, respectively. The oocysts contained approximately 40 and 50 sporocysts, respectively. The sporocysts were sub-spherical in shape and 31.2+/-2.5 x 27.0+/-2.9 microm in size. Each sporocyst contained at least 8-16 sporozoites, with the sporozoites being 12.2+/-1.4 x 3.5+/-0.5 microm in size. H. ursi n. sp. is the first Hepatozoon species recorded from the family Ursidae.

The two species common of liver fluke, Fasciola hepatica and Fasciola gigantica, cause human fascioliasis. Hybrids between these species, and introgressed forms of Fasciola, are known from temperate and subtropical regions of eastern Asia. Here, we report the presence of hybrid and/or introgressed liver flukes in Vietnam where it has recently been recognised that human fascioliasis is an important zoonotic disease. Specimens examined came from domestic stock (cattle and buffalo) at slaughter and also from human patients. DNA sequences were obtained from the nuclear ribosomal second internal transcribed spacer (ITS-2) and from portions of two mitochondrial protein-coding genes. Mitochondrial sequences in every case were similar to those of Fasciola gigantica. Nuclear ITS-2 sequences belonged to one or other of the Fasciola species, or, sequences from both were found in the same individual worm. This study extends the known range of hybrids or introgressed forms of Fasciola into tropical regions of Asia.

Adult flukes of Paragonimus species were obtained from lungs of experimental animals infected with metacercariae found in field-collected freshwater crabs in Sri Lanka. Morphological studies of adult worms under a scanning electron microscope as well as a ordinary microscope were performed in the present study. All of morphological features observed clearly indicated that this species is P. westermani. On the other hand, the shapes of metacercariae were found to be mainly oval, but semioval and spherical ones also coexisted. In spite of the variety of their morphology of the metacercariae, there is no correlation between their shapes of metacercariae and the deoxyribonucleic acid (DNA) sequences. Molecular phylogenetic analyses using two DNA regions (partial mitochondrial cytochrome c oxidase subunit 1 and second internal transcribed spacer of the nuclear ribosomal gene repeat) placed the adult flukes of P. westermani from Sri Lanka basal in a tree including all specimens of P. westermani from various areas in Asia and P. siamensis from Thailand. The present study showed that P. westermani from Sri Lanka is an ancestral form.

Arginine kinase (AK) is a member of a highly conserved family of phosphagen kinases. We determined the cDNA sequence of Toxocara canis AK, cloned it in pMAL plasmid and expressed it in Escherichia coli as a fusion protein with maltose-binding protein. The protein has a theoretical molecular mass of 45,376 Da and an estimated isoelectric point (pI) of 8.38. Alignment of the cDNA-derived amino acid sequence of T. canis AK with other phosphagen kinase sequences showed high amino acid identity with other nematode AKs, and phylogenetic analysis placed it as a distinct branch within a nematode AK cluster. Analysis of the N-terminus sequence of T. canis AK revealed the presence of a signal targeting peptide presumably targeting this protein to cytosol or endoplasmic reticulum (ER). T. canis AK showed high activity for l-arginine. The kinetic constants (K(m) = 0.12 mM, K(cat) = 29.18, and K(d) = 0.23 mM) and V(max) (43.76 micromolPi/min/mg protein) of T. canis recombinant-AK were determined for the forward reaction. It also exhibited a synergism for substrate binding (K(d)(Arg)/K(m)(Arg)=1.96). Comparison of K(cat)/K(m)(Arg) values in various arginine kinases indicates that T. canis AK has a high catalytic efficiency (248.19s(-1)mM(-1)). The present study contains the first description of arginine kinase in a zoonotic nematode. The determination of T. canis AK and its phosphagen biosynthetic pathway, which is completely different from those in mammalian host tissues, suggests this enzyme as a possible novel chemotherapy target for VLM syndrome in humans.

The aim of the present research is to determine the phylogenetic position of Setaria digitata of Sri Lanka in the evolutionary tree of filarial worms. DNA sequences of portions of the mitochondrial genes cytochrome c oxidase subunit 1 (CO1) and small subunit ribosomal RNA (12S rDNA) were analysed. Intra-specific variation was observed in CO1 but not in 12S rDNA. Phylogenetic trees inferred from these two genes resembled one another in recognizing monophyly of Setaria. S. digitata and Setaria labiatopapillosa appear to be sister species.

In the northeastern region of India, paragonimiasis is emerging as an important public health problem. However, until now the identity of the species causing human infection has been uncertain and there has been little information on the prevalence and clinicoradiological features of infection in the community. Parasitological and immunological surveys revealed that paragonimiasis was hyperendemic in parts of Arunachal Pradesh. Egg positivity in the sputum was 20.9% and 4.1% in children (age 15 years), respectively. Antibody positivity against excretory-secretory antigen of the adult worm in children and adults was 51.7% and 18.7%, respectively. Chronic cough (97.2%) and haemoptysis (83.3%) were common respiratory symptoms among egg-positive cases. Chest radiography (n=68) images from egg-positive cases showed that air space consolidation (75%), cavitary lesions (14.7%) and mediastinal adenopathy (11.8%) were very frequent. Less frequent findings were nodular lesions, bronchiectasis, mediastinal adenopathy, pleural thickening and pleural effusion. DNA extracted from eggs from the sputum of patients from Arunachal Pradesh was sequenced. Analyses of the second internal transcribed spacer (ITS2) of nuclear rDNA revealed that the species responsible is Paragonimus heterotremus.

Fifty-eight stocks of Trypanosoma cruzi from Latin America were genetically characterized using the methods of the polymerase chain reaction (PCR) and the single-stranded conformation DNA polymorphism (SSCP) with four genes, mini-exon, 24Salpha rRNA, 18Sr RNA, cruzipain, and a RAPD fragment DNA region, P7-P8. All the isolates examined were assigned to T. cruzi I or subgroups of T. cruzi II by these methods. From these results, the SSCP analysis, which was simple to perform and highly sensitive to sequence variation, seemed to be a good modality for characterizing T. cruzi, particularly for subgroups of T. cruzi II. However, in several isolates of T. cruzi II, the subgroups determined with the SSCP of 24Salpha rRNA were not consistent with those determined with other genes, the SSCP of 18S rRNA and cruzipain, and the PCR of P7-P8, possibly because of the occurrence of rare genetic exchanges or mutations or both in natural populations of this parasite. The SSCP patterns of 24Salpha rRNA and 18S rRNA were highly variable in the T. cruzi I isolates; therefore, analyses using both genes are considered to be one possible method for the characterization of isolates within T. cruzi I.

Eggs of trematode parasites are comprised of numerous vitelline cells and one fertilized ovum, and are encapsulated within a protein shell provided by the vitellocytes. In this study, we isolated two full-length cDNA clones that showed substantial levels of sequence identity with trematode-specific eggshell precursor proteins from the human lung fluke, Paragonimus westermani. These cDNAs, designated Pw-Vit20 (868-bp-long) and Pw-Vit36 (883-bp-long), shared a 76% identity with one another at the nucleotide level, and each encoded a 261-amino acid (aa) polypeptide. The deduced aa sequences contained a N-terminal hydrophobic segment, as well as a sequence motif of Gly-Gly-Gly-Tyr-Asp-Asn/Thr-Tyr-Gly-Lys/Gln, which is highly homologous with the eggshell proteins of Fasciola hepatica. With the high frequencies of tyrosine, glycine and lysine, the positions occupied by tyrosine, which has been proved to be converted into dihydroxyphenylalanine, were well preserved. Pw-Vit20 and Pw-Vit36 were found to be monoexonic genes with variably diverged variants scattered into multiple genomic loci. Their protein products were localized in the vitelline follicles and eggshells. Expression of Pw-Vit20 was restricted to the egg and adult stages, thus suggesting a critical involvement of Pw-Vit20 in the parasite's fecundity activity. Conversely, Pw-Vit36 was constitutively expressed in the metacercariae and juvenile stages in the vitelline follicles and ducts, which suggested that the prepositioning of stem or primordial vitelline cells within the juveniles prior to sexual maturation. Pw-Vit36 might acquire a unique or additional function relevant to the maturation and/or development of the vitelline cells/follicles during the evolutionary period of P. westermani. Differential biological implications of multiple eggshell precursor proteins may provide insight into the molecular mechanism of eggshell formation and the developmental process of the vitelline follicles in the parasitic trematode.

This study describes the isolation of a Toxocara canis species-specific excretory-secretory (ES) antigen and the development of an enzyme-linked immunosorbent assay (ELISA) based on this antigen. Analysis of the ES antigens of T. canis, Toxocara vitulorum, Ascaris lumbricoides and Necator americanus larval antigen was performed by SDS-PAGE followed by western blotting. A 57 kDa T. canis-specific antibody fraction (TcES-57) was identified by western blotting and labelling with anti-Toxocara antibodies (from experimental rabbits and human patients) and tracing with anti-human or anti-rabbit peroxidase conjugate. No protein fraction of 57 kDa was detected in ES or larval antigens collected from T. canis, T. vitulorum, A. lumbricoides and N. americanus. Using TcES-57, a specific antiserum was produced in rabbits and a double sandwich ELISA was developed. This test was validated using known seropositive sera from toxocariasis patients, sera from A. lumbricoides or N. americanus patients, and 50 serum samples from cats. These tests revealed that TcES-57 antigen is specific to T. canis infection and does not cross react with sera of other related infections. Thus, ELISA based on TcES-57 antigen was proven to be an effective tool in the diagnosis of toxocariasis and studies on the role of T. canis in the epidemiology of human toxocariasis.

Two cases of aberrant migration by the liver fluke Fasciola gigantica in humans are reported. In both cases, subadult worms emerged through the skin. The identity of the worms was confirmed from their DNA sequences. This uncommon human pathogen might be more likely than F. hepatica to undertake aberrant migrations in humans.

DJ-1 has recently been shown to be responsible for onset of familial Parkinson's disease (PD), PARK7. DJ-1 has been shown to play roles in transcriptional regulation and anti-oxidative stress, and loss of its function is thought to trigger onset of PD. In this study, a recombinant DJ-1 protein was administrated into the brain of PD model rats that had been injected to 6-hydroxydopamine (6-OHDA) in the left substantia nigra. PD phenotypes, including dopaminergic neuron death in the substantia nigra, decrease in dopamine, and dopamine transporter levels in the striatum, and motor abnormality, were dramatically improved by wild-type DJ-1 but not L166P DJ-1, a mutant form of DJ-1 found in PD patients. Furthermore, production of reactive oxygen species and cell death induced by 6-OHDA in SH-SY5Y cells and mesencephalic neurons were inhibited by addition of the recombinant DJ-1. These findings suggest that DJ-1 is a therapeutic target for PD.

Invasive potentials of carcinomas greatly contribute to their metastasis, which is a major threat in most cancers. We have recently shown that Arf6 plays a pivotal role in breast cancer invasive activities and identified AMAP1 as an effector of GTP-Arf6 in invasion. Expression of AMAP1 correlates well with invasive phenotypes of primary tumors of the human breast. We also have shown that AMAP1 functions by forming a trimeric protein complex with cortactin and paxillin. In this complex, AMAP1 binds to the src homology 3 (SH3) domain of cortactin via its proline-rich peptide, SKKRPPPPPPGHKRT. SH3 domains are known to bind generally to the proline-rich ligands with a one-to-one stoichiometry. We found that AMAP1/cortactin binding is very atypical in its stoichiometry and interface structure, in which one AMAP1 proline-rich peptide binds to two cortactin SH3 domains simultaneously. We made a cell-permeable peptide derived from the AMAP1 peptide, and we show that this peptide specifically blocks AMAP1/cortactin binding, but not other canonical SH3/proline bindings, and effectively inhibits breast cancer invasion and metastasis. Moreover, this peptide was found to block invasion of other types of cancers, such as glioblastomas and lung carcinomas. We also found that a small-molecule compound, UCS15A, which was previously judged as a weak inhibitor against canonical SH3/proline bindings, effectively inhibits AMAP1/cortactin binding and breast cancer invasion and metastasis. Together with fine structural analysis, we propose that the AMAP1/cortactin complex, which is not detected in normal mammary epithelial cells, is an excellent drug target for cancer therapeutics.

Paragonimus heterotremus Chen and Hsia (1964), and paragonimiasis caused by this species is a newly detected disease in Vietnam. Twelve samples of Paragonimus (Platyhelminthes: Trematoda: Digenea: Paragonimidae) from different life-stages (eggs, miracidia, metacercariae, adults from natural and experimental hosts) and host species (crab, dog, cat and human) were collected in different geographical locations in Vietnam. DNA sequences were obtained from each for partial mitochondrial cytochrome c oxidase subunit 1 (cox1) (387 bp) and the entire second ribosomal internal transcribed spacer (ITS-2) (361 bp). The ITS-2 sequences were identical among all specimens, including those previously reported in GenBank. For cox1, there were sequence differences between specimens from Vietnam (four provinces, different locations) and those from Guangxi (China) and Saraburi (Thailand). Phylogenetic trees inferred from cox1 and ITS-2 sequences using sequence data for 15 P. heterotremus and for other Paragonimus spp. revealed that all P. heterotremus originating from Vietnam, Thailand and China form a distinct group. This information also confirms the identity of the Vietnamese specimens as P. heterotremus.

A 53-year-old male patient began treatment for systemic exanthema with diaphenylsulfone (DHS) on 21 November 2002. On 18 December 2002, the patient developed a fever and additional systemic erythematous exanthema. Systemic lymphadenopathy, hepatosplenomegaly, leukocytosis (in particular, an increase in the number of atypical lymphocytes) and liver dysfunction followed. After cessation of the DHS treatment on 25 December 2002, acute renal failure occurred and the patient was transferred to Shinshu University Hospital on 4 January 2003. The patient was diagnosed with drug-induced hypersensitivity syndrome (DIHS). Steroid pulse therapy (methylprednisolone 1000 mg/day for 3 days) was given, followed by 60 mg/day of prednisolone. The patient's renal functions recovered and he was taken off hemodialysis therapy. However, the patient relapsed twice despite two sessions of steroid pulse therapy and an increase in the dose of prednisolone to 100 mg/day. Plasma exchange (PE) was carried out to reduce the activity of the disease. With a total of four plasma exchanges, we were able to reduce the dose of prednisolone from 100 mg/day to 60 mg/day without relapse. There were no adverse effects from the plasma exchanges. Plasma exchange should be considered in the treatment with corticosteroid-resistant DIHS with multiple organ lesions.

A 62-year-old woman was struck by lightning while on a mountain and fortunately did not suffer burns or unconsciousness. She stayed at a mountain lodge overnight and was taken to our hospital by helicopter the next day. Upon admission, electrocardiography showed ST segment elevation indicating acute lateral myocardial infarction, and echocardiography showed takotsubo-shaped hypokinesis of the left ventricle indicating an apical aneurysm. Her serum escaped enzyme levels were increased, as is typical in cases of myocardial infarction, however, she did not complain of cardiac symptoms. Coronary arteriography performed 4 days after admission showed a normal coronary artery while left ventriculography showed apical akinesia. An echocardiogram obtained 2 days later showed resolution of the LV wall motion abnormality. This is the first reported case of takotsubo cardiomyopathy caused by lightning. Takotsubo-shaped hypokinesis is not described as a complication of lightning-induced cardiac injury and its pathogenesis remains controversial.

DNA sequences from a portion of the mitochondrial COI gene were used to clarify phylogenetic relationships among Japanese species in the genus Cercopithifilaria. Sequences were determined from seven Japanese species, five (C. shohoi, C. multicauda, C. minuta, C. tumidicervicata and C. bulboidea) from the serow (Capricornis crispus F. Bovidae) and two (C. longa and C. crassa) from the sika deer (Cervus nippon nippon F. Cervidae). No base substitutions were observed between C. bulboidea and C. longa, suggesting that recent host switching of a lineage of C. bulboidea between bovid and cervid hosts gave rise to C. longa. In phylogenetic trees inferred using a variety of methods, the morphologically ancestral type, C. bulboidea, appeared as a derived species. C. multicauda was found to be basal in the analyses. It seems therefore that C. multicauda is the most primitive out of the seven species.