High level of IL-10 expression in the blood of animal models possibly relates to resistance against leptospirosis.

Leptospirosis is a severe zoonosis which immunopathogenesis is poorly understood. We evaluated correlation between acute form of the disease and the ratio of the anti-inflammatory cytokine IL-10 to the pro-inflammatory TNF-α and IL-1ß expression during the early phase of infection comparing resistant mice and susceptible hamsters infected with two different species of virulent Leptospira. The IL-10/TNF-α and IL-10/IL-1ß expression ratios were higher in mouse compared to hamster independently of the Leptospira strain, suggesting a preponderant role of the host response and notably these cytokines in the clinical expression and survival to leptospirosis. Using an IL-10 neutralization strategy in Leptospira-infected mouse model, we also showed evidence of a possible role of this cytokine on host susceptibility, bacterial clearance and on regulation of cytokine gene expression.

Impact of human migrations on diversity of Helicobacter pylori in Cambodia and New Caledonia.

BACKGROUND: Helicobacter pylori is a major gastric bacterial pathogen, presumed to have established itself in the human stomach approximately 100,000 years ago. Helicobacter pylori co-evolved with its host, and human migrations shaped the expansion and the diversity of strains around the world. Here, we investigated the population structure and the genomic diversity of H. pylori in New Caledonia and Cambodia, where humans of different origins are living. METHODS: Both multilocus sequence typing (MLST) and macro-array experiments were performed to assess polymorphism of housekeeping genes and to compare differences in gene contents among strains of H. pylori. RESULTS: The macro-array analysis based on variations of the flexible gene pools was consistent with the contribution of ancestral H. pylori populations to modern strains. Most of the CDS variably present encode proteins of unknown function, selfish DNA, and transposases. In New Caledonia-where humans are of several ethnic origins-strains belonged to four different genetic populations, reflecting the diversity of human populations. Melanesians and Polynesians were infected mainly by strains assigned to hspMaori, whereas Caucasians were infected by hspWAfrica, hpEurope, and hpNEAfrica strains. In contrast, strains from Khmer patients belonged to only two subpopulations: hspEAsia and hpEurope. In the two countries, both ancient and recent human migrations may have influenced the diversity of H. pylori. CONCLUSION: Our present results are consistent with the possibility of admixture of strains in multiethnic communities. This increases the global polymorphism of H. pylori without evidence of functional change or impact on fitness and virulence.

CTX-M beta-lactamases in Escherichia coli from community-acquired urinary tract infections, Cambodia.

Despite the recent global spread of CTX-M beta-lactamases in Escherichia coli isolates from community-acquired urinary tract infections (CA-UTIs), their dissemination has been little studied in developing countries. In a 2-year prospective study, we documented the prevalence of extended-spectrum beta-lactamases (ESBLs) in E. coli that were responsible for CA-UTIs in Phnom-Penh, Cambodia. Ninety-three E. coli strains were included. We observed a high prevalence of resistance to amoxicillin (88.2% of strains), cotrimoxazole (75.3%), ciprofloxacin (67.7%), gentamicin (42.5%), and third-generation cephalosporins (37.7%). A total of 34 strains carried ESBLs, all of which were CTX-M type. CTX-M carriage was associated with resistance to fluoroquinolones and aminoglycosides. U using repetitive extragenic palindromic-PCR, we identified 4 clusters containing 9, 8, 3, and 2 strains. The prevalence of CTX-M beta-lactamases has reached a critical level in Cambodia, which highlights the need for study of their spread in developing countries.

Anti-tuberculosis drug resistance and HIV co-infection in Phnom Penh, Cambodia.

The objective of this study was to observe the prevalence of drug resistance in Mycobacterium tuberculosis isolates in HIV associated tuberculosis co-infected patients in Phnom Penh City. The isolates of M. tuberculosis were collected during active laboratory-based surveillance. Of the 98 isolates studied, M. tuberculosis resistance to isoniazid was seen in 23.5%, resistance to rifampicin was seen in 16.3% and multidrug-resistance (MDR-TB) was seen in 5.1%. Our findings reveal an alarmingly high level of resistance to isoniazid and rifampicin, and confirms the need for drug susceptibility testing to guide treatment in patients with culture positive tuberculosis.

[Penicilliosis in Laos]. / Pénicilliose au Laos.

INTRODUCTION: Penicilliosis is caused by Penicillium marneffei, a dimorphic fungus endemic in southeast Asia and southern China. Since 1988, this deep mycosis has been one of the most frequent opportunistic infections in subjects with HIV infection in this region. We report the first observation of penicilliosis in Laos in a subject infected by HIV. CASE: A Laotian women with HIV infection and under treatment for tuberculosis, was hospitalized with a cough, fever and cervical lymphadenopathy. A lymph node smear stained with RAL 555 confirmed the diagnosis of penicilliosis. The histologic examination detected some Penicillium marneffei, stained by PAS and Gomori-Grocot. The patient was treated successfully with ketoconazole. DISCUSSION: Penicilliosis is endemic in southeast Asia. Especially in Thailand, it is a common opportunistic infection in HIV-infected persons. This case confirms the existence of penicilliosis in Laos, where its prevalence as an opportunistic infection is probably underestimated. The diagnostic method (slide stained with RAL 555) used in this case is simple, rapid, and appropriate for countries with limited economic resources to devote to the penicilliosis diagnosis.

[Solitary fibrous tumor of the urinary bladder]. / Tumeur fibreuse solitaire de la vessie.

INTRODUCTION: A solitary fibrous tumor is a rare circumscribed neoplasm that was long known in the pleura and has now described in many organs and soft tissues. Prognosis is favorable after complete excision. CASE: We report a solitary tumor fibrous located in the bladder of a 38-year-old man with dysuria and a palpable abdominal mass. DISCUSSION: Solitary fibrous tumors are reported very rarely in the urinary bladder. They may mimic other nonepithelial tumors in this site. Diagnosis based only on pathological features is difficult. When complete excision is possible, prognosis is generally good. Outcome is more serious in approximately 10-20% of cases, and strict follow-up is required.

Cytokine and Chemokine Expression in Kidneys during Chronic Leptospirosis in Reservoir and Susceptible Animal Models.

Leptospirosis is caused by pathogenic spirochetes of the genus Leptospira. Humans can be infected after exposure to contaminated urine of reservoir animals, usually rodents, regarded as typical asymptomatic carriers of leptospires. In contrast, accidental hosts may present an acute form of leptospirosis with a range of clinical symptoms including the development of Acute Kidney Injury (AKI). Chronic Kidney Disease (CKD) is considered as a possible AKI-residual sequela but little is known about the renal pathophysiology consequent to leptospirosis infection. Herein, we studied the renal morphological alterations in relation with the regulation of inflammatory cytokines and chemokines, comparing two experimental models of chronic leptospirosis, the golden Syrian hamster that survived the infection, becoming carrier of virulent leptospires, and the OF1 mouse, a usual reservoir of the bacteria. Animals were monitored until 28 days after injection with a virulent L. borgpetersenii serogroup Ballum to assess chronic infection. Hamsters developed morphological alterations in the kidneys with tubulointerstitial nephritis and fibrosis. Grading of lesions revealed higher scores in hamsters compared to the slight alterations observed in the mouse kidneys, irrespective of the bacterial load. Interestingly, pro-fibrotic TGF-ß was downregulated in mouse kidneys. Moreover, cytokines IL-1ß and IL-10, and chemokines MIP-1α/CCL3 and IP-10/CXCL-10 were significantly upregulated in hamster kidneys compared to mice. These results suggest a possible maintenance of inflammatory processes in the hamster kidneys with the infiltration of inflammatory cells in response to bacterial carriage, resulting in alterations of renal tissues. In contrast, lower expression levels in mouse kidneys indicated a better regulation of the inflammatory response and possible resolution processes likely related to resistance mechanisms.

Death caused by strongyloides hyperinfection in a leprosy patient on treatment for a type II leprosy reaction.

Strongyloides stercoralis is present worldwide and can cause hyperinfection in patients on long-term immunosuppressive doses of steroids, as is sometimes the case for patients treated for leprosy reactions. Strongyloides hyperinfection can present with ileus, as is discussed in this case report. Physicians, including surgeons, should be aware of this entity in order to avoid an unnecessary laparotomy. Though patients may survive if diagnosed at an early stage, strongyloides hyperinfection syndrome has a mortality rate of 87% and prevention is therefore of utmost importance.

Emergence of quinolone resistance among extended-spectrum beta-lactamase-producing Enterobacteriaceae in the Central African Republic: genetic characterization.

BACKGROUND: Cross-resistance to quinolones and beta-lactams is frequent in Enterobacteriaceae, due to the wide use of these antibiotics clinically and in the food industry. Prescription of one of these categories of antibiotic may consequently select for bacteria resistant to both categories. Genetic mechanisms of resistance may be secondary to a chromosomal mutation located in quinolone resistance determining region of DNA gyrase or topoisomerase IV or to a plasmid acquisition. The insertion sequence ISCR1 is often associated with qnr and may favour its dissemination in Gram-negative bacteria. The aim of this study was to determine the genetic mechanism of quinolone resistance among extended-spectrum beta-lactamase-producing Enterobacteriaceae strains in the Central African Republic. FINDINGS: Among seventeen ESBL-producing Enterobacteriaceae isolated from urine, pus or stool between January 2003 and October 2005 in the Central African Republic, nine were resistant to ciprofloxacin (seven from community patients and two from hospitalized patients). The ESBL were previously characterized as CTX-M-15 and SHV-12. Susceptibility to nalidixic acid, norfloxacin and ciprofloxacin, and the minimal inhibitory concentrations of these drugs were determined by disc diffusion and agar dilution methods, respectively. The presence of plasmid-borne ISCR1-qnrA region was determined by PCR and amplicons, if any, were sent for sequencing. Quinolone resistance determining region of DNA gyrase gyrA gene was amplified by PCR and then sequenced for mutation characterization. We found that all CTX-M-producing strains were resistant to the tested quinolones. All the isolates had the same nucleotide mutation at codon 83 of gyrA. Two Escherichia coli strains with the highest MICs were shown to harbour an ISCR1-qnrA1 sequence. This genetic association might favour dissemination of resistance to quinolone and perhaps other antibiotics among Enterobacteriaceae. CONCLUSIONS: This study shows that at least two mechanisms might explain the emerging resistance of Enterobacteriaceae to quinolones in the CAR. Beside the classical topoisomerase mutation, the cause may be acquisition of a plasmid-borne qnrA1. Clinicians and bacteriologists should be made aware of possible dissemination of ISCR1-qnrA1 among Enterobacteriacae.

Preliminary data about female malignant breast tumours in Cambodia.

BACKGROUND: Breast cancer is the most frequent cancer among women and the most frequent cause of cancer death in women in both developing and developed countries. However, little is known about the situation in Cambodian women living in Cambodia. OBJECTIVES: To describe the different histological types of invasive breast cancer, their frequency, grade and stage and their surgical management in Cambodia. MATERIALS AND METHODS: A retrospective study of female breast primary invasive malignant lesions processed during 2003 and 2004 in the pathology laboratory of the 'Institut Pasteur du Cambodge', in Phnom Penh, was carried out with conventional histology techniques. RESULTS: A total of 102 patients were included, with an average age of 47. Most specimens were tumour excisions or mastectomies. Tumours were generally large (37.5 % at least T3), distributed into carcinomas (94.2 %), phyllodes tumours (3.9 %) and lymphomas (1.9 %). Most cases of carcinoma presented with histological grade III (76.6%) and were invasive ductal (82.3%) or medullary (11.4%) in type. Lymphadenectomy was not systematic but nodal metastases were observed in 26 of 31 (83.9%). CONCLUSIONS: Late diagnosis of the disease is due to absence of systematic breast cancer screening and awareness. Apparent rarity of lobular carcinoma and over frequency of medullary carcinoma and phyllodes tumours should be confirmed on a larger series of patients using immunohistochemistry. Its use for other markers to help oncologists to decide whether anti-hormone treatment is indicated is also recommended, together with more standardized surgery.

Evolutionary history of Helicobacter pylori sequences reflect past human migrations in Southeast Asia.

The human population history in Southeast Asia was shaped by numerous migrations and population expansions. Their reconstruction based on archaeological, linguistic or human genetic data is often hampered by the limited number of informative polymorphisms in classical human genetic markers, such as the hypervariable regions of the mitochondrial DNA. Here, we analyse housekeeping gene sequences of the human stomach bacterium Helicobacter pylori from various countries in Southeast Asia and we provide evidence that H. pylori accompanied at least three ancient human migrations into this area: i) a migration from India introducing hpEurope bacteria into Thailand, Cambodia and Malaysia; ii) a migration of the ancestors of Austro-Asiatic speaking people into Vietnam and Cambodia carrying hspEAsia bacteria; and iii) a migration of the ancestors of the Thai people from Southern China into Thailand carrying H. pylori of population hpAsia2. Moreover, the H. pylori sequences reflect iv) the migrations of Chinese to Thailand and Malaysia within the last 200 years spreading hspEasia strains, and v) migrations of Indians to Malaysia within the last 200 years distributing both hpAsia2 and hpEurope bacteria. The distribution of the bacterial populations seems to strongly influence the incidence of gastric cancer as countries with predominantly hspEAsia isolates exhibit a high incidence of gastric cancer while the incidence is low in countries with a high proportion of hpAsia2 or hpEurope strains. In the future, the host range expansion of hpEurope strains among Asian populations, combined with human motility, may have a significant impact on gastric cancer incidence in Asia.

Expansion of European vacA and cagA alleles to East-Asian Helicobacter pylori strains in Cambodia.

Helicobacter pylori infection is associated with gastric cancer (GC). The highest incidence rates have been described in Asia, but regional variations exist that do not match the distribution of infection prevalence rates. The aim of the study was to examine the possible contribution of H. pylori virulence factors to geographic differences in the incidence of GC across East and Southeast Asia. We studied 66 isolates from Cambodian patients that had previously been assigned to two genetic populations based on sequences of seven housekeeping genes, namely hpEurope (n = 34, 51.5%) and hpEastAsia, subpopulation hspEAsia (n = 32, 48.5%). These strains were characterized with respect to vacA polymorphism and cagA status by PCR, and the CagA C-terminal region was sequenced. We also sequenced the complete cagA gene from 10 hpEurope and 10 hspEAsia strains chosen at random. The cagA gene was present in 92.4% of the 66 isolates and was mainly of Western type (n = 36, 59.0%). hspEAsia strains carrying East-Asian CagA and the m1-type vacA allele (15.2%) were less frequent among the 66 Cambodian isolates than reported in East Asian countries, a finding that might partly explain the intermediate incidence of GC in Cambodia, and by extension, in Southeast Asia (except for Vietnam). The observed high prevalence of s1a alleles (34.4%) and Western CagA (28.1%) among hspEAsia strains indicates frequent introgression of European vacA and cagA alleles into East Asian H. pylori strains. This expansion might have severe consequences for individual disease outcome.

Co-existing micronutrient deficiencies among stunted Cambodian infants and toddlers.

The prevalence of malnutrition in Cambodia is among the highest in Southeast Asia, and diarrhea and pneumonia are the leading causes of death among children. Whether these adverse health outcomes are associated with co-existing micronutrient deficiencies is uncertain. We have determined the prevalence of anaemia, as well as iron, zinc, and vitamin A deficiency and their co-existence among stunted children (77 females; 110 males) aged 6-36 mos. Non-fasting morning venipuncture blood samples were taken and analyzed for haemoglobin (Hb), serum ferritin (via IMx system), retinol (via HPLC), and Zn (via AAS), C-reactive protein (CRP) (via turbidimetry) and Hb type (AA, AE, or EE) (via Hb gel electrophoresis). Children with CRP>or=5.0 mg/L (n=34) were excluded. Zinc deficiency defined as serum Zn<9.9 micromol/L had the highest prevalence (73.2%), followed by anaemia (71%) (Hb<110 g/L), and then vitamin A deficiency (28.4%) (serum retinol<0.70 micromol/L). Of the anaemic children, only 21% had iron deficiency anaemia, and 6% had depleted iron stores. Age, log serum ferritin, and Hb type were significant predictors of Hb in the AA and AE children. Serum retinol was unrelated to haemoglobin or serum zinc. The prevalence of two or more micronutrient deficiencies (low Hb, serum retinol, and/or serum zinc) was 44%. Nearly 10% had low values for all three indices, and 18% had just one low value. In conclusion, anaemia, and deficiencies of iron, zinc, and vitamin A are severe public health problems among these stunted Cambodian children. Intervention strategies addressing multiple micronutrient deficiencies are needed.

A comparative intervention trial on fish sauce fortified with NaFe-EDTA and FeSO4+citrate in iron deficiency anemic school children in Kampot, Cambodia.

BACKGROUND: Inhabitants of agrarian villages of rural Cambodia suffer from high prevalences of iron deficiency and anemia in the context of a monotonous diet. OBJECTIVE: To compare the efficacy and safety of placebo Khmer fish sauce to that of 10 mL of fish sauce containing 10 mg of iron, added to daily school meals either as NaFe-EDTA or as FeSO4+ citrate. METHODS: 140 students aged 6-21 years were enrolled in a double-blinded, placebo-controlled intervention trial. They were randomly allocated to one of three treatment groups, and followed for 21 weeks during which 114 school meals seasoned with 10 mL of fish sauce were consumed by each participant. Changes in the concentrations of hemoglobin (hb), serum ferritin (SF), and C-reactive protein (CRP) and in body weight and standing height were determined. Prevalences of vomiting, diarrhea, and acute respiratory infections were monitored weekly. RESULTS: Both iron-fortified fish sauces increased hb and SF concentrations significantly as compared to placebo. No significant differences were observed between FeSO4+citrate and NaFe-EDTA fortification, regarding mitigation of iron-deficiency anemia (IDA) or regarding CRP, growth, infections, or side-effects. CONCLUSIONS: Iron-fortified Khmer fish sauce added to Khmer food is a suitable vehicle for iron fortification in children and adolescents. FeSO4+citrate and NaFe-EDTA show equivalent efficacy and safety.

In vitro antifungal-drug susceptibilities of mycelial and yeast forms of Penicillium marneffei isolates in Cambodia.

Penicillium marneffei infection is an important disease among human immunodeficiency virus patients in Southeast Asia. The in vitro antifungal-drug susceptibilities of 29 clinical isolates and 5 isolates from bamboo rats collected from 2002 to 2004 were determined. The P. marneffei yeast form is more susceptible than the mycelial form to amphotericin B and ketoconazole, while the mycelial and yeast forms displayed similar susceptibilities to flucytosine and itraconazole. The MICs of fluconazole were higher for both mycelial and yeast forms.

Increasing in vitro resistance to fluconazole in Cryptococcus neoformans Cambodian isolates: April 2000 to March 2002.

OBJECTIVES: Cryptococcal meningitis is the third-most-common opportunistic infection in HIV patients in Cambodia. Hospitalized patients were given amphotericin B for initial therapy followed by fluconazole for maintenance therapy. The antifungal drug susceptibility of Cryptococcus neoformans isolated from cerebrospinal fluid (CSF) was determined. METHODS: Isolates of C. neoformans were collected during active laboratory-based surveillance, the first batch from April 2000 to March 2001 (134 new cases), the second batch from April 2001 to March 2002 (268 new cases). Etest strips were used to determine the MICs of amphotericin B and fluconazole. The antigenic agglutination slide test was used for serotyping. RESULTS: The MIC(50)s and MIC(90)s of fluconazole changed significantly from year 2000 to 2002; the MIC(50)s increased from 4 to 12 mg/L, and the MIC(90)s from 12 to 96 mg/L. For amphotericin B, the MIC(50)s and MIC(90)s remained stable. Moreover, in the second batch, fluconazole MICs were >/=256 mg/L for 20 isolates. By serotyping, it was found that 98.5% of the isolates were serotype A. CONCLUSIONS: C. neoformans strains isolated from CSF of AIDS patients in Cambodia remain susceptible in vitro to amphotericin B. These strains are less susceptible in vitro to fluconazole, 2.5% being resistant in the first year and 14% in the second year of study. Nevertheless, in vitro resistance of C. neoformans to fluconazole appeared to be linked to extended maintenance treatments.