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1.
Rev Bras Parasitol Vet ; 32(4): e011923, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-38055438

RESUMO

In vitro excystation of cysts of microscopically identified Chilomastix mesnili and Retortamonas sp. isolated from Japanese macaques and Retortamonas sp. isolated from small Indian mongooses could be induced using an established protocol for Giardia intestinalis and subsequently by culturing with H2S-rich Robinson's medium supplemented with Desulfovibrio desulfuricans. Excystation usually began 2 h after incubation in Robinson's medium. DNA was isolated from excysted flagellates after 4 h of incubation or from cultured excysted flagellates. Phylogenetic analysis based on their 18S rRNA genes revealed that two isolates of C. mesnili from Japanese macaques belonged to the same cluster as a C. mesnili isolate from humans, whereas a mammalian Retortamonas sp. isolate from a small Indian mongoose belonged to the same cluster as that of an amphibian Retortamonas spp. isolate from a 'poison arrow frog' [sequence identity to AF439347 (94.9%)]. These results suggest that the sequence homology of the 18S rRNA gene of the two C. mesnili isolates from Japanese macaques was similar to that of humans, in addition to the morphological similarity, and Retortamonas sp. infection of the amphibian type in the small Indian mongoose highlighted the possibility of the effect of host feeding habitats.


Assuntos
Herpestidae , Parasitos , Retortamonadídeos , Humanos , Animais , Filogenia , Retortamonadídeos/genética , Herpestidae/genética , Macaca fuscata/genética , RNA Ribossômico 18S/genética
2.
IJID Reg ; 7: 130-135, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-37025347

RESUMO

Objectives: Trichomonas vaginalis is the most prevalent sexually transmitted parasite worldwide. However, no surveillance system exists to monitor T. vaginalis cases and drug resistance in Japan. Methods: Cervical cytology vaginal swabs were collected from women with and without suspected symptoms of T. vaginalis infection; these swabs were used for the detection of T. vaginalis, human papillomavirus (HPV), and Candida albicans using specific polymerase chain reaction. Clinical isolates of T. vaginalis were subjected to metronidazole susceptibility tests using the previously reported minimal lethal concentration (MLC) and newly established half-maximal inhibitory concentration (IC50) values. Results: The prevalence of T. vaginalis in the study population was 4.2% (5/119; 95% confidence interval [Cl], 1.5-9.7). Additionally, asymptomatic infection constituted 60% (3/5) of all cases of T. vaginalis infection. All T. vaginalis-positive patients were coinfected with HPV but not C. albicans. Five clinical T. vaginalis isolates showed metronidazole susceptibility, which was evaluated using MLC values. The quantitative IC50 values revealed that two of these clinical isolates exhibited a decreased metronidazole susceptibility. Conclusion: This is the first study to demonstrate the prevalence of T. vaginalis in Japanese women. The IC50 values of metronidazole against T. vaginalis enabled the precise and quantitative evaluation of metronidazole-susceptible T. vaginalis.

3.
Ann Vasc Surg ; 49: 313.e17-313.e19, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29455012

RESUMO

A congenital abnormality of the inferior vena cava is said to be an anatomical risk factor for venous thromboembolism. In this report, we present a case of a patient with a left duplicated common iliac vein who developed a venous thromboembolism following total abdominal hysterectomy and bilateral salpingo-oophorectomy. Only 2 items were risk factors for thromboembolism: age of ≥40 years and open surgery duration of ≥30 min; no congenital abnormalities of the inferior vena cava or thrombotic factors were observed. Thus, it was suspected that the duplicated common iliac vein could have caused the venous thromboembolism.


Assuntos
Veia Ilíaca/anormalidades , Malformações Vasculares/complicações , Trombose Venosa/etiologia , Fatores Etários , Anticoagulantes/uso terapêutico , Angiografia por Tomografia Computadorizada , Feminino , Humanos , Histerectomia/efeitos adversos , Veia Ilíaca/diagnóstico por imagem , Pessoa de Meia-Idade , Duração da Cirurgia , Flebografia/métodos , Fatores de Risco , Salpingo-Ooforectomia/efeitos adversos , Terapia Trombolítica , Resultado do Tratamento , Malformações Vasculares/diagnóstico por imagem , Trombose Venosa/diagnóstico por imagem , Trombose Venosa/tratamento farmacológico
4.
Surg Endosc ; 32(7): 3076-3086, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29313127

RESUMO

BACKGROUND: Although there have been several reports of treating large post-endoscopic submucosal dissection (ESD) ulcers by covering them with a polyglycolic acid sheet (PGAs), this approach presents problems regarding PGAs delivery. This study assessed the usefulness of a device delivery station system (DDSS) to evaluate the appropriate and rapid PGAs coating method with DDSS. METHODS: Thirty-nine of 41 patients who were diagnosed with early gastric cancer over 20 mm in diameter and pathologically diagnosed with well-differentiated adenocarcinoma were randomly allocated to the following two groups according to delivery method: the conventional PGAs delivery group (C group) (n = 19) and the new DDSS group (DDSS group) (n = 20). The primary outcome was the coating area per minute in the C group and DDSS group (cm2/min). RESULTS: There were significant differences in the coating time (min), with values of 34.1 (15.0-60.7) vs. 16.85 (11.5-27.2) min for the C group and DDSS group, respectively (p = 0.001). There was also a significant difference in coating area per minute, with values of 0.261 (0.02-1.00) and 0.96 (0.173-2.06) cm2/min for the C group and DDSS group, respectively (p = 0.001). There were four cases of post-ESD bleeding (1-7 days after ESD) in the C group compared with 0 in the DDSS group, which represented a significant difference (p = 0.030). CONCLUSIONS: The DDSS was very useful for rapidly delivering and tightly attaching a PGAs to control post-ESD bleeding. TRIAL REGISTRATION: University Hospital Medical Network (UMIN) 000026377.


Assuntos
Sistemas de Liberação de Medicamentos/instrumentação , Ressecção Endoscópica de Mucosa/efeitos adversos , Ácido Poliglicólico , Complicações Pós-Operatórias/terapia , Úlcera Cutânea/terapia , Cicatrização/efeitos dos fármacos , Adenocarcinoma/diagnóstico , Adenocarcinoma/cirurgia , Idoso , Idoso de 80 Anos ou mais , Ressecção Endoscópica de Mucosa/métodos , Desenho de Equipamento , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/etiologia , Estudos Prospectivos , Úlcera Cutânea/etiologia , Neoplasias Gástricas/diagnóstico , Neoplasias Gástricas/cirurgia , Fatores de Tempo , Resultado do Tratamento
5.
J Immunol ; 200(3): 1101-1109, 2018 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-29255076

RESUMO

Intestinal amebiasis is a major cause of diarrhea. However, research on host-amebae interactions has been hampered owing to a lack of appropriate animal models. Recently, a mouse model of intestinal amebiasis was established, and using it, we reported that Entamoeba moshkovskii colonized the intestine in a manner similar to that of the pathogenic Entamoeba histolytica In this study, we evaluated the protective mechanisms present against amebae using this model. CBA/J mice infected with E. histolytica had a persistent infection without apparent symptoms. In contrast, E. moshkovskii-infected mice rapidly expelled the ameba, which was associated with weight loss, diarrhea, and intestinal damage characterized by apoptosis of intestinal epithelial cells (IECs). Expression of NKG2D on intestinal intraepithelial lymphocytes (IELs) and IFN-γ-producing cells in Peyer's patches were significantly induced after infection with E. moshkovskii but not with E. histolytica IFN-γ-deficient mice infected with E. moshkovskii showed no obvious symptoms. Notably, none of these mice expelled E. moshkovskii, indicating that IFN-γ is responsible not only for intestinal symptoms but also for the expulsion of amebae. Furthermore, apoptosis of IECs and expression of NKG2D on IELs observed in E. moshkovskii-infected mice did not occur in the absence of IFN-γ. In vivo blocking of NKG2D in mice infected with E. moshkovskii enabled ameba to survive longer and remarkably reduced apoptotic IECs. Our results clearly demonstrate a novel protective mechanism exerted by IFN-γ against intestinal amebae, including induction of cytotoxicity of IELs toward IECs.


Assuntos
Entamoeba histolytica/imunologia , Interferon gama/imunologia , Intestinos/imunologia , Intestinos/patologia , Subfamília K de Receptores Semelhantes a Lectina de Células NK/metabolismo , Animais , Apoptose/imunologia , Modelos Animais de Doenças , Entamebíase/imunologia , Entamebíase/parasitologia , Células Epiteliais/imunologia , Interações Hospedeiro-Parasita/imunologia , Inflamação/imunologia , Interferon gama/genética , Intestinos/parasitologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , Camundongos Knockout , Nódulos Linfáticos Agregados/imunologia , Linfócitos T/imunologia
6.
Digestion ; 96(4): 239-247, 2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-29136630

RESUMO

BACKGROUND/AIMS: Multiple colorectal polyps with a diameter in the range of 10-19 mm are unable to be retrieved through a 3-mm endoscopic channel by the aspiration method. This study aims to assess the usefulness of Catcher Tag retrieval, which not only allows the accurate identification of the resected location but also enables the easiest retrieval in a short time without any special device. METHODS: One hundred thirty five patients (483 polyps) were diagnosed with colorectal neoplasm, and 64 patients (225 polyps) were enrolled and randomly allocated into the Net forceps group (NET) and the Catcher Tagged group (TAG). In TAG, 3 types of colored ring-threads were used to retrieve resected polyps. After local injection of natural saline, ring-threads were placed close to polyps. The primary outcome was the number of one-to-one correspondence locations (UMIN000020826). RESULTS: There was significant difference in one-to-one correspondence (p = 0.004). The average retrieval procedure time was 13.56 ± 3.47 (min) in NET and 3.55 ± 1.68 in TAG (p = 0.006). In NET, 1 polyp in each of 4 cases was lost during endoscopic mucosal resection and 2 polyps were lost in 1 case. In TAG, no polyp was lost (p = 0.016). CONCLUSION: The Catcher Tagged method is very useful for accurate one-to-one correspondence locations and pathological evaluation, and easy-to-retrieve multiple resected specimens.


Assuntos
Pólipos do Colo/cirurgia , Colonoscopia/métodos , Neoplasias Colorretais/cirurgia , Ressecção Endoscópica de Mucosa/métodos , Adulto , Pólipos do Colo/patologia , Colonoscopia/instrumentação , Neoplasias Colorretais/patologia , Ressecção Endoscópica de Mucosa/instrumentação , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Estudos Prospectivos , Instrumentos Cirúrgicos
7.
PLoS One ; 12(9): e0185233, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28934335

RESUMO

Our current taxonomic perspective on Entamoeba is largely based on small-subunit ribosomal RNA genes (SSU rDNA) from Entamoeba species identified in vertebrate hosts with minor exceptions such as E. moshkovskii from sewage water and E. marina from marine sediment. Other Entamoeba species have also been morphologically identified and described from non-vertebrate species such as insects; however, their genetic diversity remains unknown. In order to further disclose the diversity of the genus, we investigated Entamoeba spp. in the intestines of three cockroach species: Periplaneta americana, Blaptica dubia, and Gromphadorhina oblongonota. We obtained 134 Entamoeba SSU rDNA sequences from 186 cockroaches by direct nested PCR using the DNA extracts of intestines from cockroaches, followed by scrutinized BLASTn screening and phylogenetic analyses. All the sequences identified in this study were distinct from those reported from known Entamoeba species, and considered as novel Entamoeba ribosomal lineages. Furthermore, they were positioned at the base of the clade of known Entamoeba species and displayed remarkable degree of genetic diversity comprising nine major groups in the three cockroach species. This is the first report of the diversity of SSU rDNA sequences from Entamoeba in non-vertebrate host species, and should help to understand the genetic diversity of the genus Entamoeba.


Assuntos
Baratas/citologia , Baratas/genética , Variação Genética , Ribossomos/genética , Animais , Filogenia , Polimorfismo Genético , Análise de Sequência de DNA , Especificidade da Espécie
8.
Kansenshogaku Zasshi ; 90(1): 73-6, 2016 Jan.
Artigo em Japonês | MEDLINE | ID: mdl-27032177

RESUMO

A 66-year-old Japanese male with a history of a rectal ulcer and rectovesical fistula following brachytherapy and radiotherapy for prostate cancer, who had undergone colostomy and vesicotomy presented with a painful peristomal ulcer of approximately 5 x 2.5cm adjacent to the direction of 6 o'clock of the stoma in his left lower abdomen. Although he was admitted to be treated with intravenous antibiotics and topical debridement, the ulcer was rapidly increasing. In the laboratory findings, WBC was 12,400/µL, CRP was 16.9 mg/dL, ESR was 105mm in the first hour. Contrast enhanced CT images showed a wide high density area of skin and subcutaneous tissue around the stoma and dillitation of the transverse and descending colon. Colonoscopy showed furred profound ulcers in the rectum. A biopsy from the ulcer floor submitted to histopathology showed necrotic tissue with a mixed inflammatory infiltrates mainly composed of neutrophils and lymphocytes in the dermis. We suspected pyoderma gangrenosum with an inflammatory bowel disease in the beginning. Although he was started on oral prednisolone 60 mg daily, the ulcer did not respond to treatment. Additional methylprednisolone pulse therapy, intravenous cyclosporine and granulocytapheresis were also ineffective. A biopsy specimen from the skin ulcer margin showed erythrophagocytosis by trophozoites of amebae which were identified on PAS stained slides. The PCR method and stool examination showed positive for Entamoeba histolytica (E. histolytica), but serum antibodies were negative. Within two weeks of treatment with oral metronidazole 2,250 mg/day and topical metronidazole ointment, resolution of the ulcer was observed, then the prednisolone dosage was tapered. A split-thickness skin graft was used to cover the ulcer with a successful result. Even though we originally misdiagnosed this case, we finally reached a diagnosis of amebiasis. It is important to take account of amebiasis in the differential diagnosis of intractable ulcers which can be contaminated by feces.


Assuntos
Antibacterianos/uso terapêutico , Disenteria Amebiana/tratamento farmacológico , Entamoeba histolytica/isolamento & purificação , Úlcera/tratamento farmacológico , Idoso , Desbridamento/métodos , Combinação de Medicamentos , Disenteria Amebiana/complicações , Disenteria Amebiana/diagnóstico , Fezes/microbiologia , Humanos , Masculino , Úlcera/diagnóstico , Úlcera/etiologia
9.
Parasitol Int ; 65(5 Pt B): 520-525, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27080249

RESUMO

Entamoeba histolytica is the third leading parasitic cause of man mortality in the world. Infection occurs via ingestion of food or water contaminated with cysts of E. histolytica. Amoebae primarily colonize the intestine. The majority of amoebic infections are asymptomatic, but under some conditions, approximately 4-10% of infections progress to the invasive form of the disease. To better understand the pathogenesis of amoebiasis and the interaction between amoebae and their hosts, the development of suitable animal models is crucial. Pigs, gerbils, cats and mice are used as animal models for the study of amoebiasis in the laboratory. Among these, the most commonly used model is the mouse. In addition to intestinal amoebiasis, we developed a mouse model of liver abscess by inoculating amoeba through portal vein. However, the frequency of successful infection remains low, which is dependent on the conditions of amoebae in the laboratory. As the maintenance of virulent amoebae in the laboratory is unstable, it needs further refinement. This review summarizes mouse models of amoebiasis and the current state of laboratory culture method of amoebae.

11.
J Vet Med Sci ; 78(4): 633-40, 2016 May 03.
Artigo em Inglês | MEDLINE | ID: mdl-26685985

RESUMO

Tritrichomonas species flagellates (IMC strain) were isolated from the biliary tract of an individual who had developed cholecystitis as a complication of acquired agammaglobulinemia. Sequence analysis of Tritrichomonas sp. (IMC clone 2 (cl2)) was performed for several genetic regions including the ITS1-5.8S rDNA-ITS2 region, the cysteine protease (CP)-1, CP-2 and CP-4 to CP-9 genes, and the cytosolic malate dehydrogenase 1 gene. In addition to comparison of the variable-length DNA repeats in the isolate clone with those in T. foetus (Inui cl2) and the T. mobilensis (U.S.A.: M776 cl2) reference strains, this analysis showed that the Tritrichomonas sp. (IMC cl2) was T. foetus (cattle/swine genotype). Injection of T. foetus (IMC cl2) directly into the livers of CBA mice resulted in liver abscess formation on Day 7. Moreover, inoculation via orogastric intubation caused infection in the cecum on Day 5 in CBA mice co-infected with Entamoeba histolytica (HM-1: IMSS cl6). T. foetus (IMC cl2) was able to grow in YI-S medium for over 20 days, even at 5°C. These results indicate that the T. foetus isolate is able to survive in the feces and edible organ meat of the definitive host for a prolonged period of time, and it is possible that the parasite could infect humans.


Assuntos
Infecções por Protozoários/parasitologia , Tritrichomonas foetus/isolamento & purificação , Tritrichomonas foetus/fisiologia , Zoonoses/parasitologia , Adulto , Agamaglobulinemia/complicações , Animais , Colecistite/etiologia , Colite/parasitologia , Humanos , Masculino , Camundongos , Camundongos Endogâmicos CBA , Tipagem Molecular , Infecções por Protozoários/transmissão , Tritrichomonas foetus/classificação , Tritrichomonas foetus/genética
12.
Lung Cancer ; 75(1): 95-101, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21719142

RESUMO

Expression of the transcription factor FOXP3 characterizes regulatory T cells (Tregs) that engage in the maintenance of immunological self-tolerance and immune homeostasis. Intra-tumoral accumulation of Tregs is associated with unfavorable prognosis in several kinds of cancers. Recently, expression of FOXP3 and its association with prognosis have also been shown in some cancer cells in clinical studies. For non-small cell lung cancer (NSCLC), however, prognostic significance of tumor FOXP3 expression and its relationship with Tregs remain unknown. FOXP3 expression in cancer cells and tumor-infiltrating lymphocytes was examined by immunohistochemical staining of surgical specimens from 87 patients with NSCLC. Prognostic values of the tumor-infiltrating Treg count and tumor FOXP3 expression status were evaluated retrospectively. FOXP3-positive cancer cells were observed in 27 of 87 (31.0%) patients. There was no significant relationship between Treg count and tumor FOXP3 status. Increased Treg counts were associated with worse overall and relapse-free survival whereas the influence of tumor FOXP3 status on survival was not significant. However, when FOXP3-positive cancer cells were present, the relationship between Treg accumulation and worse prognosis was attenuated. In contrast, patients without tumor FOXP3 expression and high Treg count had significantly worse overall and relapse-free survival (hazard ratio: 3.118 and 3.325, p=0.028 and 0.024, respectively) than other groups. These results suggest that tumor FOXP3 expression has a better prognostic potential in NSCLC and that in combination with tumor-infiltrating Treg count the absence of tumor FOXP3 allows the selection of high-risk patients.


Assuntos
Carcinoma Pulmonar de Células não Pequenas/imunologia , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Fatores de Transcrição Forkhead/biossíntese , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/metabolismo , Linfócitos do Interstício Tumoral/imunologia , Linfócitos T Reguladores/imunologia , Idoso , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Intervalo Livre de Doença , Feminino , Proteína Forkhead Box O3 , Fatores de Transcrição Forkhead/genética , Fatores de Transcrição Forkhead/imunologia , Humanos , Imuno-Histoquímica/métodos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Linfócitos do Interstício Tumoral/metabolismo , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Prognóstico , Recidiva , Linfócitos T Reguladores/metabolismo
13.
Parasitol Res ; 110(6): 2095-104, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22179263

RESUMO

Cell motility by actin cytoskeleton is essential for differentiation processes of excystation and encystation of Entamoeba. We recently studied an actin depolymerizing factor (ADF)/cofilin (Cfl) of Entamoeba invadens (Ei), and demonstrated its contribution to the encystation and excystation of E. invadens through actin cytoskeletal reorganization. Profilin is also an actin-binding protein but its function is different from that of Cfl in actin assembly. This study investigated E. invadens profilins in relation to encystation and excystation which were induced in axenic culture systems. A homology search of the E. invadens genome database and molecular cloning identified four profilins of the parasite named EiPFN1, EiPFN2, EiPFN3, and EiPFN4. There were also multiple genes of profilin in Entamoeba histolytica (Eh) and Entamoeba dispar (Ed), each of which had three profilins. A search for conserved domains revealed that these profilins of Entamoeba had actin, phosphoinositide, and poly-proline binding sites. Phylogenetic analysis revealed that EiPFN3 and EiPFN4 formed the same clades including EhPFN3 and EdPFN3, and EhPFN2 and EdPFN2, respectively, while EiPFN1 and EiPFN2 were separated from EhPFN1 and EdPFN1. Rabbit anti-EiPFN1 serum reacted with recombinant EiPFN3 and EiPFN4 but not EiPFN2, and also reacted with EiPFN in lysates of cysts and trophozoites. Immunofluorescence staining with this antiserum showed co-localization of EiPFN with actin beneath the cell membrane through the life stages and also showed cytoplasmic localization. Both proteins proved to be rich in pseudopodia of trophozoites. Real-time RT-PCR showed that the mRNA level of EiPFN1 and EiPFN4 in trophozoites was comparable but that of EiPFN2 and EiPFN3 was very low. During encystation, the mRNA expression of EiPFN1 and EiPFN4 increased remarkably in the early phase much higher than that of EiPFN2 and EiPFN3. Then, the expression of all four PFNs sharply decreased in the later phase. This was in contrast to the sharp decrease in the mRNA level of EiCfl-2 during encystation in our previous study. In cysts, EiPFN1 was most abundantly expressed and EiPFN4 was at a lower level, while the expressions of EiPFN2 and EiPFN3 were virtually absent. Following the induction of excystation, mRNA levels of EiPFN1, EiPFN2, and EiPFN4 in cysts 5 h after induction were significantly higher than those in cysts before induction, while that of EiPFN3 was slightly higher than before induction. The mRNAs of EiPFN1 increased most extensively when the excystation was induced in the presence of cytochalasin D. Small interfering RNA (siRNA) to EiPFN1 inhibited both encystation and excystation but not growth. These findings demonstrate different expression of EiPFNs and the contribution of EiPFN to the encystation and excystation.


Assuntos
Entamoeba histolytica/crescimento & desenvolvimento , Entamoeba histolytica/patogenicidade , Perfilação da Expressão Gênica , Profilinas/genética , Animais , Anticorpos Antiprotozoários/imunologia , Sítios de Ligação , Membrana Celular/química , Citoplasma/química , Entamoeba histolytica/genética , Imunofluorescência , Microscopia Confocal , Dados de Sequência Molecular , Filogenia , Coelhos , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos
14.
Kyobu Geka ; 64(6): 445-9, 2011 Jun.
Artigo em Japonês | MEDLINE | ID: mdl-21682039

RESUMO

OBJECTIVE: The aim of this study was to reveal the clinicopathological feature of granulocyte colony-stimulating factor (G-CSF) producing lung cancer. METHOD: Nine cases of G-CSF producing lung cancer from July 2003 to July 2008 were retrospectively evaluated. RESULTS: All cases were male, 8 cases were poorly differentiated carcinoma. Average of leucocyte and serum G-CSF were 23,378/microl and 128.6 pg/ml respectively. Five cases had febrile symptom, average of serum C-reactive protein (CRP) was 13.37 mg/dl. Immunohistological examination showed positive staining for G-CSF in 6 cases. Serum interleukin-6 (IL-6) level was elevated in 3 cases. Clinical stages were IB in 2, IIB in 2, IIIA in 3 and IIIB in 2 patients. Chemotherapy was performed for patients with stage IIIB. Operation was performed for the other cases. Five cases were died within 12 months, whereas 4 cases are surviving for 6 to 16 months. CONCLUSION: Generally, the prognosis of G-CSF producing lung cancer seems to be poor, but in our institute there were 2 cases who lived over 1 year without disease. It is important to establish more effective adjuvant therapy for G-CSF producing tumor.


Assuntos
Fator Estimulador de Colônias de Granulócitos/biossíntese , Neoplasias Pulmonares/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Humanos , Masculino , Pessoa de Meia-Idade , Prognóstico
15.
Parasitol Res ; 109(2): 417-23, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21286750

RESUMO

Entamoeba histolytica forms chitin-walled cysts during encystation process, where formation of the cyst wall needs not only chitin synthase but also chitinase. During excystation, quadruplet amoebae emerge from the chitin-walled cysts by dissolving the wall, so that chitinase may be necessary for excystation process as well. There is, however, no report on chitinase expression during excystation. In this study, we used Entamoeba invadens, a reptilian amoeba, as a model for encystation and excystation of E. histolytica, and studied chitinase mRNA expression in those processes. Although expression of three E. invadens chitinases designated EiChit1, EiChit2, and EiChit3 during encystation has been reported, we identified another enzyme named as EiChit4 in the E. invadens genome database. Therefore, we investigated the primary structure and mRNA expression of these four chitinases of Ei in the excystation as well as the encystation by real-time reverse transcription polymerase chain reaction (RT-PCR). Like EiChit1, EiChit4 had an 8 × Cys chitin-binding domain (CBD) and a hydrophilic spacer between the CBD and catalytic domain, and was also closer to EiChit1 than EiChit2 and EiChit3 in the phylogenetic tree. During encystation, the expression of all four chitinases increased in the early phase; the increase in EiChit1 and EiChit4 was much higher than in EiChit2 and EiChit3. Then, the expression of all four chitinases sharply decreased in the later phase. In cysts, EiChit1 was most abundantly expressed and EiChit4 was at a lower level, while the expressions of EiChit2 and EiChit3 were virtually absent. Following the induction of excystation, mRNA levels of EiChit1 and EiChit4 in cysts 5 h after induction were significantly lower than those in cysts before induction, while those of EiChit2 and EiChit3 were remarkably higher than before induction. The mRNAs of only EiChit2 and EiChit3 remarkably increased when the excystation was induced in the presence of cytochalasin D. These data demonstrate different structures and expressions of four chitinases in the differentiation of E. invadens.


Assuntos
Quitinases/biossíntese , Quitinases/genética , Entamoeba/enzimologia , Entamoeba/crescimento & desenvolvimento , RNA Mensageiro/biossíntese , Esporos de Protozoários/enzimologia , Esporos de Protozoários/crescimento & desenvolvimento , Sequência de Aminoácidos , Análise por Conglomerados , DNA de Protozoário/química , DNA de Protozoário/genética , Entamoeba/genética , Entamoeba/metabolismo , Regulação da Expressão Gênica , Dados de Sequência Molecular , Filogenia , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Esporos de Protozoários/genética , Esporos de Protozoários/metabolismo
16.
Exp Parasitol ; 127(1): 195-201, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-20691182

RESUMO

The differentiation processes of excystation and encystation of Entamoeba are essential for infection and completion of their life-cycle, and the processes need cell motility and its control by actin cytoskeletal reorganization. This study investigated actin depolymerizing factor (ADF)/cofilin (Cfl) family proteins, which are important molecules in actin cytoskeletal reorganization, in Entamoeba invadens in relation to the encystation and excystation. Axenic culture systems were used to induce encystation and excystation. A homology search of the E. invadens genome database and molecular cloning identified three ADF/Cfl family proteins of the parasite (named for short as EiCfl-1, EiCfl-2, and EiCfl-3). This is different from other Entamoeba species, i.e. Entamoeba histolytica and Entamoeba dispar, each of which has only one ADF/Cfl family protein. These ADF/Cfl of E. invadens do not have Ser3 (serine locates third from first methionine), similar to E. histolytica, E. dispar, Saccharomyces cerevisiae and Schizosaccharomyces pombe, although the activity of ADF/Cfl is negatively regulated by phosphorylation of the Ser3 in metazoans. Phylogenetic analysis revealed that Entamoeba Cfl formed a distinctive clade that is separate from other organisms, and the branches of the tree were separated in two consistent with the presence and absence of Ser3. Rabbit anti-EiCfl-2 serum reacted with all recombinant EiCfls and EiCfl in lysates of cysts, trophozoites and metacystic amoebae. Immunofluorescence staining with this antiserum showed co-localization of EiCfl with actin beneath the cell membrane through the life stages. Both proteins proved to be rich in pseudopodia of trophozoites and metacystic amoebae. Real-time RT-PCR showed that mRNAs of EiCfl-2 and actins were highly expressed, but there were few mRNA of EiCfl-1 and EiCfl-3. Remarkably decreased mRNA levels were observed in EiCfl-2 and actins during encystation. All three EiCfls and actins became transcribed after the induction of excystation. The mRNAs of only EiCfl-1 and EiCfl-3 increased remarkably when the excystation was induced in the presence of cytochalasin D. These findings demonstrate that EiCfl-2 and actins co-localize beneath the cell membrane in trophozoites and cysts as well as metacystic amoebae being rich in pseudopodia, that EiCfl-1 and EiCfl-3 are expressed only after the induction of excystation, and that enhanced excystation by cytochalasin D is associated with high expression of EiCfl-1 and EiCfl-3.


Assuntos
Fatores de Despolimerização de Actina/metabolismo , Entamoeba/fisiologia , Fatores de Despolimerização de Actina/química , Fatores de Despolimerização de Actina/genética , Fatores de Despolimerização de Actina/imunologia , Sequência de Aminoácidos , Animais , Clonagem Molecular , Citocalasina D/farmacologia , DNA de Protozoário/química , Entamoeba/química , Entamoeba/classificação , Entamoeba/genética , Expressão Gênica , Soros Imunes/imunologia , Inibidores da Síntese de Ácido Nucleico/farmacologia , Filogenia , Reação em Cadeia da Polimerase , RNA Mensageiro/metabolismo , RNA de Protozoário/genética , RNA de Protozoário/isolamento & purificação , Coelhos , Alinhamento de Sequência
18.
Kyobu Geka ; 63(10): 875-8, 2010 Sep.
Artigo em Japonês | MEDLINE | ID: mdl-20845697

RESUMO

Pulmonary hamartoma is most common benign tumor of the lung and is not recognised as having a character of malignant transformation. So, longtime radiological observation is not uncommon for patients with diagnosis of pulmonary hamartoma from computed tomography (CT) finding. Although pulmonary hamartoma does not transform to malignancy, high frequency of coexistence hamartoma and lung cancer has been reported. We experienced 14 cases of resected pulmonary hamartoma, and 3 of them had lung cancer, showing that 21.4% of pulmonary hamartoma coexisted with lung cancer. Patients with pulmonary hamartoma should undergo sufficient evaluations for malignancy.


Assuntos
Adenocarcinoma/patologia , Hamartoma/patologia , Neoplasias Pulmonares/patologia , Neoplasias Primárias Múltiplas/patologia , Humanos , Masculino , Pessoa de Meia-Idade
19.
Microbiol Immunol ; 54(7): 425-33, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20618689

RESUMO

The clinical features of PCP differ according to the factors responsible for the predisposing immunosuppression. Although the diagnosis of PCP often requires BAL, the profiles of the inflammatory mediators in the BAL fluid are not thoroughly documented. The aim of the current study was to characterize the profiles of inflammatory mediators in BAL fluid during PCP in patients with underlying autoimmune diseases, malignancies, or AIDS. The medical records of 14 patients with autoimmune diseases, 10 with malignancies, and 8 with AIDS, all of whom had been diagnosed with PCP by microscopic examination of BAL fluid, were reviewed. The concentrations of TNF-alpha, MCP-1, HMGB1, IL-8, IL-6, IL-10, and IFN-gamma in the BAL fluid that had been obtained for the diagnosis of PCP were measured. The concentrations of MCP-1, IL-8, and IL-6 differed according to the underlying disease, tending to be higher in patients with autoimmune diseases and lower in those with AIDS. The concentrations of HMGB1, IL-8, and IL-6 were positively correlated with the proportion of neutrophils in BAL fluid and inversely with the oxygenation index. Although the serum concentrations of CRP and LDH were positively correlated with those of IL-8 and MCP-1, none of the mediators in BAL fluid was correlated with the serum beta-D-glucan concentration. The production of inflammatory mediators in the lung differed between the patient groups with different underlying disorders. The modest upregulation of IL-8 and IL-6 might be associated with the milder clinical manifestations of PCP in AIDS patients.


Assuntos
Líquido da Lavagem Broncoalveolar/química , Citocinas/análise , Mediadores da Inflamação/análise , Pneumonia por Pneumocystis/imunologia , Síndrome da Imunodeficiência Adquirida/complicações , Síndrome da Imunodeficiência Adquirida/imunologia , Adulto , Idoso , Doenças Autoimunes/complicações , Doenças Autoimunes/imunologia , Líquido da Lavagem Broncoalveolar/imunologia , Citocinas/imunologia , Feminino , Humanos , Mediadores da Inflamação/imunologia , Pulmão/química , Pulmão/imunologia , Masculino , Pessoa de Meia-Idade , Pneumonia por Pneumocystis/diagnóstico , Estudos Retrospectivos
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