In gene therapy trials, adeno-associated virus (AAV) vectors are injected directly into target tissues such as muscle and liver. Direct injection can lead to the introduction of a low level of the vector into blood circulation. To determine the systemic effects of the vector released in the blood, we extensively examined the biodistribution of intravenously administered AAV serotype 2 (AAV2) vector in cynomolgus monkeys. Although the vector distribution pattern varied from monkey to monkey, the vector DNA was maintained in the various tissues beyond 7 months post-inoculation (pi). The vector DNA was detected in the lymphoid tissues, particularly in the spleen, more frequently and at a much higher level than in the other tissues tested (i.e., brain, lung, liver, heart, gallbladder, pancreas, colon, kidney, ovary, uterus, etc.). The expression of a transgene was detected in the lymph nodes at 3 months pi. The distribution of two pseudotyped vectors, AAV2/10 and AAV2/11, was similar to that of the AAV2 vector. The present results suggest that when introduced intravenously, the AAV vector DNA persists and may induce transgene expression in various monkey tissues. Thus, the possibility of inadvertent gene transfer to various non-target tissues should be considered in a gene therapy strategy with an AAV vector.

In March 2004, we experienced an outbreak of Chlamydia pneumoniae infection on an islet of Korea. In order to assess the significance of the epidemic, we performed a mass examination of 137 students (7 - 16 years old; male, 69; female, 58) at a school. The examination consisted of a questionnaire inquiring about respiratory symptoms, a serum antibody test for C. pneumoniae using a microimmunofluorescence (MIF) method and enzyme-linked immunosorbent assay (ELISA), and nasopharyngeal swab tests to detect of the organism by specific PCR and cell culture. The results demonstrated that 72 (58.3%) of the students had respiratory symptoms such as rhinorrhea, a sore throat, and/or cough or fever. The PCR positivity of acute-phase patients was 63% (12/19) and PCR positivity using the culture sample was 94% (18/19). However, the existence of the organism was not confirmed fluorescein isothiocyanate (FITC). ELISA, one of the serological methods utilized, demonstrated, in the same patients, 48% (13/27) positive IgM antibodies at the acute phase of the outbreak, and 16% (3/19) positive IgM antibodies during the convalescent phase. The index value (ID) 3.0 for single-sera IgG was 19% (5/27) and that for IgA was 4% (1/27) at the acute phase; the corresponding percentages in the convalescent phase were 11% (2/19) and 5% (1/19), respectively. However, as regards paired sera, no patient demonstrated a 1.35 ELISA ID value at 2 weeks, or an increased value of 1.0 at 8 weeks after the onset of the outbreak. In the MIF experiment, the percent positivity of unpaired IgM from the acute phase was 58% (11/19). At the convalescent phase, this percentage was 47% (9/19); however, the positivity of paired serum IgG was 26% (5/19). In the same sample, the percentage of positive cases demonstrated by both ELISA and MIF approaches for single IgM was 37% (7/19) at the acute phase and 11% (2/19) at the convalescent phase. We were unable to isolate C. pneumoniae by cell culture, but we did obtain sufficient serological and PCR data to consider C. pneumoniae as the causative agent of the outbreak. Meaningful results were acquired in terms of serology, and were compared to the healthy population in Korea. Although it remains necessary to investigate the possibility of co-infection and to determine whether or not this outbreak coincides with the prevalence of influenza, it was unequivocally concluded that this outbreak of C. pneumoniae infection has occurred on an islet of Korea.

Streptococcus pneumoniae susceptibility to 12 antimicrobial agents was assessed using isolates collected from patients with invasive and non-invasive infections in a Spanish medical center. Two hundred and thirty-six invasive and 478 non-invasive pneumococcal isolates obtained between 1998 and 2004 were tested. Penicillin non-susceptible isolates were more likely to exhibit resistance to cephalosporins, macrolides, chloramphenicol, and tetracycline when compared to penicillin-susceptible isolates. Penicillin resistance was present in 8.1% of the invasive and 18.6% of the non-invasive isolates. Overall, antimicrobial resistance was greater in non-invasive versus invasive isolates in adults. Serogroups included in the 7-valent and 23-valent formulation accounted for approximately 92.8 and 88.3% of the invasive isolates in children 2 years old or younger and the elderly, respectively. The proportion of isolates not susceptible to penicillin, erythromycin, and/or tetracycline decreased significantly over the surveillance period. Local epidemiological data assisted in the clinical determination of treatment protocols and effective prevention strategies.

Intestinal microsporidiosis is most commonly associated with persistent diarrhea in advanced AIDS cases. To determine the prevalence and clinical manifestations of this infection in HIV/AIDS patients, a single fresh stool sample and blood were collected from 243 (214 HIV-positive and 29 HIV-negative) diarrheal patients. The presence of intestinal microsporidiosis in the stool was determined by Uvitex-2B staining and a PCR-based detection method. HIV screening was done by using ELISA, and reactive samples were confirmed by Western blotting. The CD4+ cell count was analyzed using FACScan. Out of 243 diarrheal patients, 39 (16.0%) cases were positive for intestinal microsporidial infection by either of the methods used. Of the 39, only 18 cases positive by microscopy were also positive by PCR. Based on PCR and microscopic analyses the microsporidial parasites were identified as Enterocytozoon bieneusi (30), Ecephalitozoon intestinalis (6), and double infections (3). All microsporidia-positive cases were HIV-positive, and 92.3% had diarrhea for over 4 weeks. The diarrhea was watery in 79.5% of the patients. Weight loss >10% was recorded in 37 (94.9%) cases. The CD4+ cell count was <100 cells/mm³ in 84.4% of subjects, and 59.4% of the patients had a CD4+ cell count of ≤50 cells/mm³, with a mean of 22.8 cells/mm³. This study revealed that intestinal microsporidiosis is a common cause of chronic diarrhea and severe weight loss in advanced AIDS patients in Ethiopia. This condition is attributable mainly to E. bieneusi. Thus, early diagnosis of intestinal microsporidiosis in HIV/AIDS patients would certainly be helpful in the understanding and management of diarrheal illness.

We compared changes in the relative frequencies of opportunistic illnesses (OIs) among 1,044 Taiwanese persons with HIV infection who were enrolled in three study periods from June 1994 to June 2004: before the introduction of highly active antiretroviral therapy (HAART); early HAART; and late HAART. Nearly two-thirds of those newly diagnosed with HIV infection had CD4 counts below 200/mm³ . Among a total of 1,182 OIs, candidiasis, Pneumocystis carinii pneumonia, and tuberculosis were the most common in all three study periods within 3 months after enrollment. Within 3 to 12 months after enrollment, the number of OIs declined significantly. Only 81 cases of OIs were diagnosed, the most common of which were cytomegalovirus disease, tuberculosis, candidiasis, and Mycobacterium avium complex infection. Except for the decreasing frequency of Kaposi’s sarcoma, there was no significant change in the relative frequency of each individual OI within 3 months and within 3 - 12 months after enrollment across the three study periods. We concluded that despite the significant decrease in the number of AIDS-related OIs, the relative frequencies of common OIs did not change significantly after the introduction of HAART.

Subacute sclerosing panencephalitis (SSPE) can show variations in its clinical course. Typical initial symptoms consist of intellectual decline, deterioration in school performance, and myoclonic jerks. Atypical forms of presentation such as generalized seizures and acute or subacute coma can be observed rarely. This report describes a 5-year-old boy with an atypical feature of SSPE, generalized seizures. For 1 month his only symptom was seizures, followed by personality and behavioral changes and myoclonic jerks. A diagnosis of SSPE was made based on the boy’s history of measles, characteristic electroencephalogram changes and compatible magnetic resonance, and elevated anti-measles antibody titers in the cerebrospinal fluid and serum. The case presented in this article is a good example of SSPE in which, at early stages, some of the signs and symptoms can lead to an erroneous diagnosis.

We experienced a case of a 76-year-old man who developed a splenic abscess while undergoing treatment for interstitial pneumonia. Splenic abscess-like abnormal intensities were accidentally found by the chest computed-tomography (CT) examinations 3 weeks after the initiation of corticosteroids and immunosuppressive treatment for interstitial pneumonia. An ultrasonography-guided percutaneous aspiration test resulted in the isolation of methicillin-resistant Staphylococcus aureus (MRSA). Since colonized MRSA had been detected intermittently from sputum after admission and the patient risked bloodstream infection from an indwelling central venous catheter and intubation, we suspected that the organism colonized in the airway had spread into the bloodstream via these devices. Although CT-guided percutaneous drainage followed by postoperative antibiotic therapy are normally required for the treatment of splenic abscess, the patient was successfully treated by the administration of vancomycin without drainage.

There have recently been large outbreaks of Marburg hemorrhagic fever (MHF) caused by Marburgvirus (MARV) in the Democratic Republic of Congo and Angola. The development of reliable diagnostic systems for MHF is urgently needed. An antigen-capture enzyme-linked immunosorbent assay (Ag-capture ELISA) using either of the two monoclonal antibodies (2A7 and 2H6) produced by immunizing mice with recombinant nucleoprotein of MARV was described (Journal of Medical Virology, 76, 111-118, 2005). In the present study, it was revealed that the Ag-capture ELISA specifically detected authentic MARV antigen and that the sensitivity of the Ag-capture ELISA was at a level similar to that of reverse-transcription polymerase chain reaction. These results suggest that the Ag-capture ELISA using the monoclonal antibodies, 2A7 and 2H6, is applicable to the diagnosis of MHF.

During the years 2000 to 2004, of 248 serologically confirmed cases of Crimean-Congo hemorrhagic fever (CCHF) that occurred in several parts of Iran, 169 were reported from Sistan-va-Baluchestan province. To assess the seroprevalence of CCHF virus infection within the Zahedan and Zabol districts of the Sistan-va-Baluchestan province in Iran, 300 subjects were sampled from the general population. In addition to blood sampling, a questionnaire was completed for every subject. All but just 3 of our 300 sampled subjects participated in blood sampling, and just 7 out of the 297 serum samples were found to be IgG ELISA positive. The point estimate of the seroprevalence was 0.024 (95% confidence interval: 0.003 - 0.044). A history of keeping livestock in houses (even for short periods) showed an association with seropositivity (P = 0.018). It seems that even occasional contact with livestock could be effective in transmission of the virus.

Fluoxetine (FLX), a P-glycoprotein inhibitor with antimalarial activity, is a promising candidate for reversing chloroquine/mefloquine (CQ/MQ) resistance. The Dd2 strain of CQ- and MQ-resistant Plasmodium falciparum was synchronized and assayed with various concentrations of CQ/MQ individually and in combination with FLX or verapamil (VPL). Our results indicated the 50% inhibitory concentration values of CQ and MQ were greatly lowered when FLX was used simultaneously. Isobolograms indicated that CQ-FLX combinations are more synergistic (mean fractional inhibitory concentration [FIC] index 0.55) than MQ-FLX (mean FIC index 0.64), and their synergistic effect was better than or at par with CQ-VPL (mean FIC index 0.88) and MQ-VPL (mean FIC index 0.60) combinations. We conclude that the FLX potentiates the CQ and MQ response on multidrug-resistant P. falciparum at clinically achievable concentrations.

To clarify the location of pulmonary granulomas in vivo, we prepared a Mycobacterium tuberculosis H37Rv mutant in which the gene for a green fluorescent protein (GFP) (GFP-H37Rv) was introduced. Five weeks after aerosol infection with GFP-H37Rv, the infected lungs from guinea pigs and mice were subjected to imaging using a photon imager. Pulmonary granulomas more than 1 mm in diameter were localized clearly by the photon imager. Therefore, if a method for binding a dye (GFP, fluorescein isothiocyanate [FITC], etc.) specifically to M. tuberculosis can be developed, it will be possible to visualize granulomas using a photon imager.

There has been worldwide resurgence in the incidence of Streptococcus pyogenes infection and its sequelae. S. pyogenes remains uniformly susceptible to penicillin, and it is speculated that its minimum inhibitory concentration (MIC) has not changed during the past 70 years. The purpose of the present study was to determine the occurrence and pattern of resistance to penicillin and erythromycin amongst clinical isolates of S. pyogenes. A total of 34 clinical strains of S. pyogenes were identified by standard procedures. Antimicrobial susceptibility was analyzed by the Kirby-Bauer method of disk diffusion, and the E-test method was used to determine the MIC to penicillin and erythromycin. All the strains were sensitive to penicillin, clindamycin and vancomycin on disk diffusion. Ten (29.4%) strains were resistant to erythromycin. The pattern of macrolide resistance observed was M type. By the E-test method, 7 (20.6%) strains were penicillin nonsusceptible and 6 (17.6%) were erythromycin resistant. We concluded that surveillance of its susceptibility pattern is crucial to monitoring the development of antibiotic resistance in S. pyogenes.

Mycobacterium marinum is one of the nontuberculosis mycobacteria responsible for skin infections. There have been very few case series of M. marinum infections reported in the English literature. Herein, we describe three patients with M. marinum tenosynovitis. All patients had positive cultures and were exposed to pricking by a fishbone. The incubation period ranged from 7 to 60 days. Key elements in the diagnosis of this infection were a high index of suspicion raised by negative bacterial tissue cultures, poor response to conventional antibiotics treatment, a history of exposure to tropical fish and tissue biopsy for culture and histology. The treatment is essentially antimicrobial therapy supplemented by an appropriate surgical debridement, especially when deep structures are involved.

The purpose of this study is to investigate the distribution and structure of Salmonella genomic island 1 (SGI1) among Salmonella enterica serovar Derby isolates from swine and their rearing environment. Three variants of SGI1s, specifically SGI1-A, C, and I, were identified by PCR mapping. The results of macro-restriction analysis and DNA sequencing of SGI1 flanking regions revealed that there are at least two genomic lineages of Derby strains bearing SGI1s.