Genome-wide Analysis of Chlamydophila pneumoniae Gene Expression at the Late Stage of Infection

Chlamydophila pneumoniae, an obligate intracellular eubacterium, changes its form from a vegetative reticulate body into an infectious elementary body during the late stage of its infection cycle. Comprehension of the molecular events in the morphological change is important to understand the switching mechanism between acute and chronic infection, which is deemed to relate to the pathogenesis of atherosclerosis. Herein, we have attempted to screen genes expressed in the late stage with a genome-wide DNA microarray, resulting in nomination of 17 genes as the late-stage genes. Fourteen of the 17 genes and six other genes predicted as late-stage genes were confirmed to be up-regulated in the late stage with a quantitative reverse transcriptase–polymerase chain reaction. These 20 late-stage genes were classified into two groups by clustering analysis: ‘drastically induced’ and ‘moderately induced’ genes. Out of eight drastically induced genes, four contain σ28 promoter-like sequences and the other four contain an upstream common sequence. It suggests that besides σ28, there are certain up-regulatory mechanisms at the late stage, which may be involved in the chlamydial morphological change and thus pathogenesis

シンニュウセイ オ タイショウ トシタ キツエン ボウシ キョウイク シコウ ガ タバコ ニ タイスル イシキ ニ アタエル エイキョウ ノ ケントウ

大学入学早期からの喫煙防止教育の大切さがこれまでに指摘されていたが,喫煙防止教育の効果に関してほとんど検討されていない.本研究は,大学新入学時に喫煙防止教育を施行し,その前後の喫煙に対する態度や喫煙防止教育を行う意欲や自信の変化等を検討することを目的とした.2008 年4 月に,新入生74 名(男子54 名,女子20 名)を対象に喫煙防止教育を施行し,教育の直前とその一週間後に同一アンケート調査を行い,PASW Statistics 18.0 for Windows を使用し,c 2 検定及びt 検定を行った.喫煙防止教育後の喫煙者の禁煙に対する関心度や非喫煙者の喫煙に対する関心度には,有意差は見られなかった.さらに,喫煙防止教育を受けた経験が十分あるかという質問に関しては,「タバコの害」と「タバコの依存性」では教育前と有意差は無く,「禁煙指導の方法」に関してのみ,有意に上昇した.しかし,喫煙行動に対する態度に関しては,「医療施設の全面禁煙化」以外の全項目で有意に上昇した.さらに,タバコに関する社会の動きに対する関心度が有意に高まり,喫煙防止教育に対する意欲や自信も有意に上昇した.本研究は長期的な教育効果を検討するには不十分であるが,喫煙防止教育として新入生の意識向上に良い影響を与えた.Several studies have discussed the importance of educatingstudents on smoking prevention during the initial yearsof university education. In this study we examined thechanging perceptions of smoking behavior among freshmenin university just before and after 1 week of education onthe health hazards due to smoking.In April, 2008, 74 freshmen (54 boys and 20 girls) of aparamedical university were provided health education onthe hazards caused by smoking, and the changes in the perceptionsregarding the smoking behavior were examinedby the basis of a questionnaire that was answered by thestudents before and after the education;the same questionnairewas filled both the times. For statistical examination,c 2 test and t test were performed using IBM SPSS(PASW Statistics 18.0) for Windows.Even after educating the students on smoking prevention,there was no significant difference in the attitude ofsmokers toward smoking cessation and that of non-smokerstoward smoking. Although the awareness among thestudents with regard to the health hazards of smoking wasnot significantly different before and after this education,their awareness of the methods to quit smoking had increasedsignificantly. Moreover, the mean values of the differentopinions on the smoking behavior increased significantly.However, one exception was the opinions in themedical institutions that had tobacco-free environments.The mean values that increased the highest after this education,were "Implementation of non-smoking in the entireuniversity" and "The need for effective education that assistsin quitting smoking." Further, after this education, thescore of interest in social activities against smoking increased.Education on anti-smoking is vital for freshmenafter enrollment in a university

Liver Transcriptome Profiles Associated with Strain-Specific Ehrlichia chaffeensis-Induced Hepatitis in SCID Mice ▿ †

Infection of humans with Ehrlichia chaffeensis, the etiologic agent of human monocytic ehrlichiosis, can cause hepatitis of various levels of severity. When the three human isolates of E. chaffeensis, each belonging to a different genogroup, are inoculated into severe combined immunodeficiency mice, the order of severity of clinical signs and bacterial burden detected in the liver is as follows (from greatest to least severity and highest to lowest burden): strain Wakulla, followed by strain Liberty, followed by strain Arkansas. In this article, we used microarray analysis to define transcriptional profiles characteristic of the histopathological features in the mouse liver. Cytokine and chemokine profiles and their receptor profiles were strikingly different among the three strains of E. chaffeensis: gamma interferon, CCL5, CXCL1, CXCL2, CXCL7, CXCL9, interleukin 2 receptor gamma (IL2Rγ), IL21R, CCR2, and CXCR6 were highly upregulated with strain Arkansas; and tumor necrosis factor (TNF), CCL2, CCL3, CCL5, CCL6, CCL12, CCL20, CXCL2, CXCL7, CXCL9, CXCL13, TNF receptor superfamily 9 (TNFRSF9), TNFRSF13β, IL1R2, IL2Rγ, IL20Rβ, IL21R, CCR1, CCR2, and CXCR4 were highly upregulated with strain Wakulla. With strain Liberty, only CXCL13 was highly upregulated, and IL13Rα2 was downregulated. In livers infected with the Arkansas strain, monocytes/macrophages and NK cells were enriched in the granulomas and an increase in NK cell marker mRNAs was detected. Livers infected with the Wakulla strain displayed infiltration of significantly more neutrophils and an increase in neutrophil marker mRNAs. Genes commonly upregulated in liver tissue infected with the three strains are other host innate immune and inflammatory response genes, including those encoding several acute-phase proteins. Genes downregulated commonly are related to host physiologic functions. The results suggest that marked modulation of host cytokine and chemokine profiles by E. chaffeensis strains underlies the distinct host liver disease

Virulence Potential of Ehrlichia chaffeensis Strains of Distinct Genome Sequences▿ †

Human monocytic ehrlichiosis, one of the most frequent life-threatening tick-borne zoonoses, is caused by Ehrlichia chaffeensis that lacks endotoxin and peptidoglycan. While sequence polymorphisms in several genes in E. chaffeensis strains have been reported, global genomic divergence and biological differences among strains are unknown. The objectives of the present study were to compare the genome sequences of strains of E. chaffeensis and to examine the virulence potentials of the strains with defined genome sequences. Genomic DNA was extracted from purified E. chaffeensis strains Wakulla and Liberty, and comparative genome hybridization was performed using a densely tiled microarray of 147,027 chromosome positions of the E. chaffeensis strain Arkansas genome. The results revealed that 4,663 and 5,325 positions in the chromosomes of strains Wakulla and Liberty, respectively, were different from those in the chromosome of strain Arkansas, including three common major polymorphic chromosomal regions. Of various functional categories, the differences were most concentrated in genes predicted to encode cell envelope proteins. Of all the open reading frames (ORFs), 21 omp-1 (p28 gene) paralogs, nine genes encoding hypothetical proteins, two genes encoding ankyrin repeat proteins, and hemE contained the most differences. Several highly polymorphic ORFs were confirmed by sequencing. When the E. chaffeensis strains were inoculated into severe combined immunodeficiency mice, the order of the severity of clinical signs and the bacterial burden detected in mice was Wakulla > Liberty > Arkansas. Severe diffuse inflammation and granulomatous inflammation were evident in the livers of mice infected with strains Wakulla and Arkansas, respectively, but not in the livers of mice infected with strain Liberty. These results revealed distinct virulence phenotypes of E. chaffeensis strains with defined genome sequences

Four VirB6 Paralogs and VirB9 Are Expressed and Interact in Ehrlichia chaffeensis-Containing Vacuoles▿ †

The type IV secretion system is an important virulence factor in several host cell-associated pathogens, as it delivers various bacterial macromolecules to target eukaryotic cells. Genes homologous to several virB genes and virD4 of Agrobacterium tumefaciens are found in an intravacuolar pathogen Ehrlichia chaffeensis, the tick-borne causative agent of human monocytic ehrlichiosis. In particular, despite its small genome size, E. chaffeensis has four tandem virB6 paralogs (virB6-1, -2, -3, and -4) that are 3- to 10-fold larger than A. tumefaciens virB6. The present study for the first time illustrates the relevance of the larger quadruple VirB6 paralogs by demonstrating the protein expression and interaction in E. chaffeensis. All four virB6 paralogs were cotranscribed in THP-1 human leukemia and ISE6 tick cell cultures. The four VirB6 proteins and VirB9 were expressed by E. chaffeensis in THP-1 cells, and amounts of these five proteins were similar in isolated E. chaffeensis-containing vacuoles and vacuole-free E. chaffeensis. In addition, an 80-kDa fragment of VirB6-2 was detected, which was strikingly more prevalent in E. chaffeensis-containing vacuoles than in vacuole-free E. chaffeensis. Coimmunoprecipitation analysis revealed VirB9 interaction with VirB6-1 and VirB6-2; VirB6-4 interaction with VirB6-1, VirB6-2, and VirB6-3; and VirB6-2 80-kDa fragment interaction with VirB6-3 and VirB6-4. The interaction of VirB9 and VirB6-2 was confirmed by far-Western blotting. The results suggest that E. chaffeensis VirB9, the quadruple VirB6 proteins, and the VirB6-2 80-kDa fragment form a unique molecular subassembly to cooperate in type IV secretion

FIGURE 7 in Genetic and morphological variation analyses of Glandirana rugosa with description of a new species (Anura, Ranidae)

FIGURE 7. Dorsal (A), lateral (B), and ventral (C) views and the oral disc (D) of a tadpole of Glandirana reliquia sp. nov. in stage 35 of Gosner (1960), collected on 9 May 2013 at Sendai City, Miyagi Pref., Japan. Scale bar = 10 mm.Published as part of Shimada, Tomohiko, Matsui, Masafumi, Ogata, Mitsuaki, Miura, Ikuo, Tange, Mai, Min, Mi-Sook & Eto, Koshiro, 2022, Genetic and morphological variation analyses of Glandirana rugosa with description of a new species (Anura, Ranidae), pp. 25-45 in Zootaxa 5174 (1) on page 38, DOI: 10.11646/zootaxa.5174.1.2, http://zenodo.org/record/697293

FIGURE 8 in Genetic and morphological variation analyses of Glandirana rugosa with description of a new species (Anura, Ranidae)

FIGURE 8. Advertisement call of Glandirana reliquia sp. nov. from Tokyo, Japan, recorded at an air temperature of 20.1°C, showing sonogram (top) and wave form (bottom).Published as part of <i>Shimada, Tomohiko, Matsui, Masafumi, Ogata, Mitsuaki, Miura, Ikuo, Tange, Mai, Min, Mi-Sook & Eto, Koshiro, 2022, Genetic and morphological variation analyses of Glandirana rugosa with description of a new species (Anura, Ranidae), pp. 25-45 in Zootaxa 5174 (1)</i> on page 38, DOI: 10.11646/zootaxa.5174.1.2, <a href="http://zenodo.org/record/6972938">http://zenodo.org/record/6972938</a&gt

Genetic and morphological variation analyses of Glandirana rugosa with description of a new species (Anura, Ranidae)

Shimada, Tomohiko, Matsui, Masafumi, Ogata, Mitsuaki, Miura, Ikuo, Tange, Mai, Min, Mi-Sook, Eto, Koshiro (2022): Genetic and morphological variation analyses of Glandirana rugosa with description of a new species (Anura, Ranidae). Zootaxa 5174 (1): 25-45, DOI: https://doi.org/10.11646/zootaxa.5174.1.