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Sequencing of a new target genome: the Pediculus humanus humanus (Phthiraptera: Pediculidae) genome project.

The human body louse, Pediculus humanus humanus (L.), and the human head louse, Pediculus humanus capitis, belong to the hemimetabolous order Phthiraptera. The body louse is the primary vector that transmits the bacterial agents of louse-borne relapsing fever, trench fever, and epidemic typhus. The genomes of the bacterial causative agents of several of these aforementioned diseases have been sequenced. Thus, determining the body louse genome will enhance studies of host-vector-pathogen interactions. Although not important as a major disease vector, head lice are of major social concern. Resistance to traditional pesticides used to control head and body lice have developed. It is imperative that new molecular targets be discovered for the development of novel compounds to control these insects. No complete genome sequence exists for a hemimetabolous insect species primarily because hemimetabolous insects often have large (2000 Mb) to very large (up to 16,300 Mb) genomes. Fortuitously, we determined that the human body louse has one of the smallest genome sizes known in insects, suggesting it may be a suitable choice as a minimal hemimetabolous genome in which many genes have been eliminated during its adaptation to human parasitism. Because many louse species infest birds and mammals, the body louse genome-sequencing project will facilitate studies of their comparative genomics. A 6-8X coverage of the body louse genome, plus sequenced expressed sequence tags, should provide the entomological, evolutionary biology, medical, and public health communities with useful genetic information.

Animals↗

Host associations and seasonal occurrence of Haemaphysalis leporispalustris, Ixodes brunneus, I. cookei, I. dentatus, and I. texanus (Acari: Ixodidae) in Southeastern Missouri.

Hemaphysalis leporispalustris (Packard), Ixodes brunneus Koch, Ixodes cookei Packard, Ixodes dentatus Marx, and Ixodes texanus Banks were collected during a 3-yr study of pathogen-tick-host interactions in southeastern Missouri. H. leporispalustris was collected from the eastern cottontail rabbit, Northern bobwhite, and Carolina wren, and it was active all year. I. brunneus was collected by drag and from passerine birds during December, March, and April. I. cookei was collected from raccoons and mink during April, June, September, October, and November. I. dentatus was collected from the cottontail rabbit and Carolina wren throughout the year. I. texanus was collected from the eastern gray squirrel, Virginia opossum, and raccoon throughout the year.

Animals↗

Genome-wide transcriptional landscape of Mycobacterium tuberculosis during acute lung infection.

Tuberculosis (TB) remains a major global health burden, yet the mechanisms by which Mycobacterium tuberculosis (Mtb) adapts to host environments to drive disease pathology are incompletely defined. A key limitation has been reliance on axenic culture systems that fail to recapitulate the complex, host-imposed stresses encountered by Mtb in vivo. Here, we report the first microarray-based genome-wide transcriptomic profiling of Mtb in rabbit lungs with active TB, which closely mirrors human disease features, including granuloma heterogeneity, necrosis, and cavitation. Using Mtb RNA isolated from infected lung homogenates or broth-culture, we capture bacterial transcriptional states shaped by the host microenvironments. The transcriptional data analyses reveal extensive, context-dependent reprogramming of Mtb metabolic, respiratory, and stress-response networks that diverges markedly from in vitro expression profiles, including activation of stress adaptation, lipid catabolism, nucleic acid metabolism, and transcriptional regulation pathways. These data uncover pathways and networks that are selectively engaged in vivo and likely critical for Mtb survival within granulomatous lesions. Our findings demonstrate that transcriptional states most relevant to TB pathogenesis are underrepresented in standard lab-grown Mtb models and highlight the importance of in vivo bacterial profiling. By characterizing Mtb gene expression within diseased lungs, this study provides a systems-level framework for understanding TB pathogenesis and reveals in vivo-essential pathways, offering potential targets for translational drug discovery and the development of more effective anti-TB therapies.

Animals↗

[In vitro infection by different strains of Paracoccidioides brasiliensis].

We analyzed the in vitro infection process by P. brasiliensis and the effect of extracellular factor(s) produced on monolayers of mammalian Vero cell lines. The yeast phase of four strains was studied: B339 (avirulent or slightly virulent), U, (intermediate virulence), 93745 and 63265 (both highly virulent). Strains of intermediate and high virulence had higher adherence at first contact (about 16%). Strain B339 had a slower adherence at first contact (8%) than the others during the same period. The production of extracellular proteases, soluble extracellular factor(s) and extracellular antigen gP43 showed no correlation with the in vitro physiopathogenicity of the analyzed strains. We demonstrate that the Vero model presented in this paper is a suitable system to study infection and virulence in vitro. We are currently assessing its usefulness as a tool for the analysis of the interaction between pathogen, host and antifungal agents.

Animals↗

Predicting antibacterial response from pharmacodynamic and pharmacokinetic profiles.

The aim of antibacterial chemotherapy is to achieve sufficient drug concentrations at the site of infection for an adequate length of time to ensure bacterial eradication and optimize clinical success. Whether the desired outcome is achieved or not depends on a number of pathogen-, drug- and patient-related factors. Neither microbiologic activity nor antibacterial pharmacokinetic data alone can adequately describe the complex interaction between pathogen, host and antibacterial during the disease process. A relatively new discipline - pharmacodynamics - seeks to integrate both microbiologic and pharmacokinetic data. The particular model that best predicts clinical outcome depends on the pattern of microbial killing and the persistence of antibacterial effects after plasma concentrations have fallen below the minimum inhibitory concentration (MIC) for the target pathogen (post-antibiotic effect [PAE]). The beta-lactams, for example, exhibit time-dependent bacterial killing with minimal persistent effects. Time above MIC (T(MIC)) is therefore the parameter that best correlates with clinical efficacy for these agents and that, in turn, necessitates multiple daily dosing to optimize the duration of exposure. The macrolides erythromycinA and clarithromycin exhibit a similar pharmacokinetic/pharmacodynamic relationship to that of the beta-lactams, although for clarithromycin the area under the concentration-time curve (AUC) also correlates with clinical outcome (reflecting the more prolonged PAE of this agent). Azithromycin, ketolides, such as telithromycin (HMR 3647), streptogramins and fluoroquinolones exhibit concentration-dependent killing and have prolonged persistent effects, such that the AUC:MIC or Cmax:MIC ratio correlates most closely with clinical efficacy. For these agents the aim is to maximize drug concentrations to which the target pathogen is exposed and this may require higher doses and hence enable longer dosing intervals to be used. In summary, pharmacodynamic models provide a unique approach to determining likely in vivo activity of individual antibacterial agents and prediction of clinical outcomes.

Animals↗

[Characteristics of immune response in acute intestinal enterobacterial infections].

Update literature on the problem of microorganism--host interactions with pathogenic enterobacteria as a model is reviewed. The activation of lymphoid cells and their protective role is shown to be due to the stimulating action of bacterial antigens. The role of cytokines and stimulation of their expression are discussed. Some phenomena of immunosuppression, phases of enteric mucosal immune response are described.

Animals↗

Using a cDNA microarray to study cellular gene expression altered by Mycobacterium tuberculosis.

OBJECTIVE: To examine the global effects of Mycobacterium tuberculosis (M. tuberculosis) infection on macrophages. METHODS: The gene expression profiling of macrophage U937, in response to infection with M. tuberculosis H(37)R(a), was monitored using a high-density cDNA microarray. RESULTS: M. tuberculosis infection caused 463 differentially expressed genes, of which 366 genes are known genes registered in the Gene Bank. These genes function in various cellular processes including intracellular signalling, cytoskeletal rearrangement, apoptosis, transcriptional regulation, cell surface receptors, cell-mediated immunity as well as a variety of cellular metabolic pathways, and may play key roles in M. tuberculosis infection and intracellular survival. CONCLUSIONS: M. tuberculosis infection alters the expression of host-cell genes, and these genes will provide a foundation for understanding the infection process of M. tuberculosis. The cDNA microarray is a powerful tool for studying pathogen-host cell interaction.

Cells, Cultured↗

[Proliferation and apoptosis in primary gastrointestinal B-cell non-Hodgkin s lymphoma and its association with Helicobacter pylori].

OBJECTIVE: To investigate primary gastrointestinal B-cell non-Hodgkin s lymphoma for clinicopathological features, proliferation, apoptosis and its association with Helicobacter pylori (HP). METHODS: Classification of tumors, expression of HP, proliferation and apoptosis related gene products were studied by immunohistochemistry. Apoptosis was studied by TUNEL (TdT [terminal deoxynucleotidyl transferase] dUTP nick end-labeling). RESULTS: There were 15 cases of mucosa associated lymphoid tissue (MALT) lymphoma (9 in stomach, 6 in intestine) and 42 cases of diffuse large B cell lymphoma (DLBCL) (28 in stomach, 14 in intestine) in all the 57 cases of primary gastrointestinal BCL. The average apoptotic indexes (AI) were 0.16%, 2.54% in MALT lymphoma and DLBCL respectively. The average proliferation indexes (PI) were 2.22% and 8.71%,respectively. The p53 positive rates were 6.7% and 35.7%. PI (P=0.026) and p53 (P=0.044) had significant differences; But Bcl-2 positive rates were 60.0% and 35.7%,and HP positive rates were 66.7% and 23.8%, respectively. HP had significant differences (P=0.005). CONCLUSION: HP and gastrointestinal MALT lymphoma had significant relation. Apoptosis and proliferative activity were higher in DLBCL than in MALT. The higher Bcl-2 expression in MALT may be related to lower apoptosis. p53 promotes apoptosis and p53 gene mutation may play a role in the progression from MALT to DLBCL. HP antibody may be used in HP detection in gastrointestinal BCL.

Adolescent↗

[Cytomegalovirus infection of myeloma cell line KM3 and its effect on IL-6 mRNA expression in the cells].

OBJECTIVE: To investigate the infection of myeloma cell line KM3 by cytomegalovirus (CMV) as well as its effect on IL-6 mRNA in the cells. METHODS: RT-PCR assay was used to detect the mRNA expression of CMV immediate early gene (IE), glyceraldehyde phosphate dehydrogenase (GAPDH) and IL-6 of KM3 cells infected by 100-, 10-, 1-folds of median tissue culture infective dose (TCID(50)) of CMV. Flow cytometry was used to detect the expression of pp65 antigen. CMV particles were detected with electron microscope. RESULTS: KM3 cells infected by CMV could express IE mRNA as compared with the uninfected cells. CMV pp65 antigen positive cells were (5.58 +/- 1.55)% and (3.75 +/- 0.85)% respectively when infected by 100 and 10 TCID(50) of CMV, which were significantly higher than that of control group (P < 0.05). CMV particles could be seen in the infected cells. Expression of IL-6 mRNA was increased after the infection. CONCLUSION: CMV can infect myeloma cell line KM3 and replicate in the cells. CMV can increase the expression of IL-6 mRNA.

Cell Line, Tumor↗

[In vitro infection of human megakaryocyte precursors by human cytomegalovirus (HCMV) and the antiviral effect of HCMV antisense oligonucleotides].

OBJECTIVE: To explore the suppression effect of human cytomegalovirus (HCMV) on megakaryocytes and their precursors and study the antiviral effect of antisense phosphorothioate deoxyoligonucleotide (ASON) against HCMV. METHODS: CD34(+) cells were induced to proliferate and differentiate committedly to megakaryocytes in a semi-solid CFU-MK culture system. Cultured cells and ASON pretreated CD34(+) cells were infected by HCMV of AD169 strain. HCMV immediate early protein (IEP) DNA and mRNA and UL36 mRNA were detected by PCR and reverse transcription-polymerase chain reaction (RT-PCR). Cytotoxicity was evaluated by MTT assay. RESULTS: HCMV AD169 suppressed the proliferation of megakaryocytes significantly. Compared with the mock group, the CFU-MK yields were decreased by 21.6%, 33.8%, and 46.3%, respectively, in 3 different titers of virus infected groups (P < 0.05). The suppression was virus titer dependent. HCMV IEP DNA, HCMV IEP mRNA and UL36 mRNA were detected in the colony cells of viral infection group. Compared with the infected group by HCMV AD169, UL36Anti treatment at 0.08 micromol/L could recover the CFU-MK yields significantly (P < 0.05). In the infected MK, which was pretreated with UL36Anti at 0.08 micromol/L, HCMV UL36 mRNA was undetectable by RT-PCR. The oligonucleotide MM(1) containing a G-to-C substitution in UL36Anti was inactive at 0.08 micromol/L but active at 0.40 micromol/L. The concentration of UL36Anti necessary to significantly affect cell growth was 90.00 micromol/L. CONCLUSIONS: HCMV AD169 infection inhibits the proliferation and differentiation of megakaryocytes and their precursors. There are early transcriptions of HCMV IE and UL36 protein in infected CFU-MK. The specific ASON has a definite anti-HCMV activity.

Antigens, Viral↗

[The relationship between activities of chitinase and beta-1,3-glucanase and resistance to rhizomania in sugar beet].

The relationship between activities of chitinase and beta-1,3-glucanase and resistance to rhizomania of sugar beet were studied in diseased soil and disease-free soil by using a method of comparative physiology. Two resistant varieties and two susceptible ones of sugar beet were used in this experiment. Chitinase and beta-1,3-glucanase activities of the resistant varieties were higher than the susceptible ones in either diseased soil or disease-free soil. There is a positive correlation between chitinase, beta-1,3-glucanase activity and yield of beet in diseased soil, suggesting that pathogen infection can induce resistance to rhizomania through activating chitinase and beta-1,3-glucanase.

Beta vulgaris↗

[O2-* burst of tobacco leaves triggered by Erwinia carotovora subsp. carotovora inoculation].

O(-.)(2) production based on chemical method by Ecc-treated intact tobacco leaves was observed. It showed a long-lasting one-phase course beginning 1 h and ending 14 h after Erwinia carotovora subsp. carotovora (Ecc) was inoculated. In Ecc-treated leaves, O(-.)(2) production rate of 3 h is 2 times of 1 h, and of 10 h it remains 1.7 times. It decreases rapidly between 10 h and 14 h, at 14 h it falls to the level before inoculation. This is a completely different course from the widely accepted two-phase course obtained from suspension-cultured cells. Electron spin resonance (ESR) spectrum of O(-.)(2) produced by intact tobacco leaves after Ecc inoculation 2 h and 6 h was also observed, the amplitude of 6 h is larger than of 2 h in Ecc-treated group. In the control group, there is no difference between the amplitude of 2 h and 6 h. This indicated that O(-.)(2) amount of Ecc-treated group is higher than of the control group, which is the same result as obtained through chemical method. The ESR spectrum of O(-.)(2) produced by chloroplasts and mitochondria from tobacco leaves after 2 h and 6 h after Ecc inoculation were observed. In chloroplast experiment, the amplitude of 6 h was larger than of 2 h in Ecc-treated group while it was the same in the control group. ESR spectrum of O(-.)(2) produced by mitochondria was proved to be a same result after a careful comparison, in spite of a greater difference between 2 h and 6 h in control group than chloroplast experiment. The fact that spectra of these two organelles were both synchronous with that of intact leaves implied that these two organelles possibly participated in cellular O(-.)(2) burst. Chloroplasts in the dark showed no ESR spectrum 2 h and 6 h after Ecc inoculation in Ecc-treated group as well as in the control group, indicating that O(-.)(2) in the chloroplast probably originated from the photosynthetic electron transport process.

Chloroplasts↗

[Detection of autoimmune parameter of SARS patients].

OBJECTIVE: To explore whether autoimmune phenomena exist in SARS patients, and to seek for unusual autoimmune antibodies in SARS patients. METHODS: Autoantibodies against cell nuclei (ANA), autoantibodies against smooth muscles (SMA), autoantibodies against parietal cells (PCA), autoantibodies against heart cells (HRA) were detected by using immunofluorescence, and autoantibodies against live-kidney microsomes (LKM) and anti-M2 antibodies were detected by ELISA in sera taken from 27 SARS patients and 18 healthy controls. Immunofluorescence was used to localize the targets antigens in slides with biochips of lung (monkey) of SARS associated antibodies. RESULTS: ANA, AMA, LKM and SMA were found positive in 3, 1, 1, and 1 of the 27 SARS sera. In 18 healthy control sera, one ANA and one AMA were positive. Statistical analysis showed that there were no difference between two groups in every item detected. Twenty-six of 27 SARS patients and 5 of 18 healthy controls had strongly stained columnar epithelia of the bronchiole, especially the lumen border of the epithelia?the difference between two groups was significant. CONCLUSION: No antibodies against organs but lung were found in SARS patients. There are auto antibodies against lung tissues in sera of SARS patients.

Adolescent↗

[Effects of HCMV on phenotypes of parotid duct epithelial cells and its mechanisms].

OBJECTIVE: To investigate the effects of HCMV infection on phenotypes of parotid duct epithelial cells and relative mechanisms. METHODS: The expressions of immediate early antigen of HCMV, pan cytokeratin and cathepsin D etc. were detected by immunohistochemical staining in tissues of parotid cytomegalic inclusion disease. RESULTS: Cytokeratin which acts as an epithelial marker became negative while staining of Cathepsin D was intensified in parotid duct epithelial cells after infected by HCMV. CONCLUSION: It demonstrated that cytokeratin was lost through over-expression of Cathepsin D in parotid duct epithelial cells infected by HCMV.

Animals↗

[Relations between the expression of cyclin E, p16ink4, ki67 and HPV16/18 infection in cervical exfoliated cells].

OBJECTIVE: To confirm the relations between the expression of cyclin E, p16ink4, ki67 and HPV16/18 infection using cervical exfoliated cells, and evaluate the usefulness of cyclin E, p16ink4 and ki67 as biomarkers for screening of cervical carcinomas. METHODS: The expression of cyclin E, p16ink4 oncoproteins and ki67 proliferative activity was evaluated immunohistochemically in 78 cervical exfoliated epithelial specimens. Human papillomavirus type16 and 18 (HPV16/18) infection was assessed by polymerase chain reaction (PCR) using type specific primers. RESULTS: Cyclin E, p16ink4 and ki67 were all overexpressed in cervical preneoplasia and neoplasia cells, compared with little expressed in ASCUS (P less than 0.005). Overexpression of cyclin E was observed in CIN, (P less than 0.01), p16ink4 and ki67 overexpressed in invasive carcinoma(100 percent and 90.9 percent respectively). The degree of p16ink4 and ki67 expression correlated well with the degree of cervical neoplasia (P less than 0.005). HPV16 infection was assessed at all stages of cervical neoplasia samples, and a significant relationship with the degree of cervical epithelial lession was observed at the same time. The expression level of p16ink4 and ki67 seemed to be more closely associated with HPV16 infection than cyclin E did (rs=1.0 vs rs=0.4). HPV18 was found positive in only 1 case in CIN1 and in 4 cases in CIN2-3. Therefore no significance was found on statistical analysis (P less than 0.005). CONCLUSION: Cyclin E, p16ink4 and ki67 should be regarded as useful biomarkers of HPV-related cervical neoplasias, and be used for screening patients at high risk for developing cervical carcinomas. Moreover, cyclin E might be a significant cytologic marker for the primary screening of cervical carcinomas.

Adult↗

[Changes of serum soluble P-selectin and tumor necrotic factor-alpha in patients with CMV induced acute coronary syndrome].

OBJECTIVE: To explore the changes and relationship between serum soluble P-selectin, tumor necrosis factor-alpha (TNF-alpha) in coronary heart disease patients with acute coronary syndrome (ACS) and human cytomegalovirus (HCMV) infection. METHODS: The levels of circulating soluble P-selectin, TNF-alpha, HCMV-IgM and HCMV-IgG were determined by enzyme-linked immunosorbent assay (ELISA) in 79 cases for ACS group, 30 cases for stable angina (SA) group and 30 healthy control cases. RESULTS: (1) The serum positive rate of HCMV-IgM and HCMV-IgG in the ACS, SA and healthy control groups were 30.4% (24/79), 10.0% (3/30) and 6.7% (2/30); 86.1% (68/79), 80.0% (24/30) and 53.3% (16/30), respectively. Positive rate of HCMV-IgM in the ACS was higher than those in SA and healthy control groups (P < 0.01), positive rate of HCMV-IgG in the ACS and SA groups were higher than that of the healthy control group (P < 0.01). (2) Compared with the SA group and healthy control group, the levels of the serum soluble P-selectin and TNF-alpha were significantly higher in patients with ACS [(6437.3 +/- 666.9) pg/ml vs. (1520.0 +/- 112.7) pg/ml and (1481.0 +/- 109.1) pg/ml, (56.2 +/- 18.4) pg/ml vs. (27.3 +/- 13.7) pg/ml and (28.1 +/- 11.3) pg/ml], respectively, P < 0.01). The AMI group, compared with the UA group in the ACS group, had significantly higher levels of the serum soluble P-selectin and TNF-alpha (P < 0.01). Compared with the SA group, the levels of the serum soluble P-selectin and TNF-alpha were not significantly different in healthy control group. (3) The levels of the serum soluble P-selectin and TNF-alpha in HCMV-IgM positive patients were significantly higher than the HCMV-IgM negative patients in the ACS group (P < 0.01). CONCLUSION: The chronic infection with HCMV might injure endothelial cells that subsequently contribute to the formation and progression of atherosclerosis, the acute infection with HCMV may induce increased serum levels of soluble P-selectin and TNF-alpha that might participate in acute coronary events.

Acute Coronary Syndrome↗

[Immunohistochemical evidence for HIV-1 infection in the liver of HIV-infected patients].

OBJECTIVE: To detect p24 antigen of human immunodeficiency virus (HIV)-1 in the liver biopsy specimens of patients with HIV infection. METHODS: Liver biopsy samples from 14 patients with HIV/AIDS (11 man, 3 women; age range 27-52 years; infection time range 8-13 years) were examined by immunohistochemistry prospectively. RESULTS: Intracellular expression of HIV-1 p24 antigen was detected in Kupffer cells, endothelial cells and hepatocytes. There were more HIV-positive liver cells in the patients with severer liver damage than those with milder liver damage (t=2.5189, P=0.0270). CONCLUSION: These findings indicate that HIV-1 could replicate in the liver of HIV-infected patients and might be related to the liver cells apoptosis.

Adult↗

[Relationship between core promoter mutation, clinical features and virus replication in HBV carriers].

OBJECTIVE: To elucidate the relationship between HBV core promoter mutation and clinical features as well as its effects on serum e system and viral replication. METHODS: Semi-nested mutation specific PCR (msPCR) was employed for detecting core promoter mutation at nt 1 762-1 764 in 97 patients with HBV infection. RESULTS: The msPCR method was demonstrated to be specific and reliable for the mutation detection by sequencing the PCR products. The detection ratio of the mutation in patients with acute hepatitis, mild, moderate and severe chronic hepatitis and liver cirrhosis was 2/5, 7/43, 10/31, 1/3 and 7/15, respectively. The detection rate of the mutation in liver cirrhosis was significantly higher than that in light chronic hepatitis (P < 0.025). In 92 patients with chronic HBV infection, HBeAg positive rate in wild (25/92), mutant (42/92) and mixed (25/92) strain infection was 80.0%, 56.0% and 64.3%, HBV DNA level was (4.4 +/- 8.5) x 10(8), (1.1 +/- 1.6) x 10(9) and (1.4 +/- 1.8) x 10(9) copies/ml, the rate of abnormal ALT was 44.0%, 52.0% and 42.6%; ALT level was (58.6 +/- 79.0), (57.1 +/- 75.2) and (62.6 +/- 90.3) IU/L, respectively (P > 0.05). CONCLUSIONS: The msPCR method for detecting core promoter mutation at nt 1 762-1 764 is specific and reliable. Core promoter mutation is associated with the severity of liver disease, but neither related to the status of e system in serum nor to the virus replication.

Adolescent↗