Distribution of fluconazole-resistant Candida bloodstream isolates among hospitals and inpatient services in Israel.

The emergence of fluconazole-resistant Candida (FRC) is worrisome, but little is known about susceptibility patterns in different nosocomial settings. We prospectively analysed Candida bloodstream isolates in 18 medical centres in Israel (six tertiary-care and 12 community hospitals). The study included 444 episodes of candidaemia (450 patient-specific isolates, 8.5% fluconazole-resistant). Institutional FRC bloodstream infection rates correlated with annual inpatient days, and were strongly associated with the presence and activity of haematology/oncology services. Infection with Candida krusei and fluconazole-resistant Candida glabrata occurred exclusively in hospitals with >600 beds. These findings suggest that empirical antifungal strategies should be tailored to the nosocomial setting.

Isolation of mesenchymal stem cells from G-CSF-mobilized human peripheral blood using fibrin microbeads.

Adult mesenchymal stem cells (MSC) that are able to differentiate into various mesenchymal cell types are typically isolated from bone marrow, but their significant presence in human peripheral blood (PB) is controversial. Fibrin microbeads (FMB) that bind matrix-dependent cells were used to isolate MSC from the mononuclear fraction of mobilized PB of adult healthy human donors treated with a granulocyte colony-stimulating factor. Isolation by plastic adherence resulted in a negligible number of MSC in all samples tested, whereas FMB-based isolation yielded spindle-shaped cell samples that could further expand on plastic or on FMB in eight out of the 11 samples. The yield of these cells at days 17-18 after the harvest was approximately 0.5% of the initial cell number. The isolated cells were grown on plastic and characterized by FACS analysis and immunohistochemistry for specific markers. Following culturing and first passage, the FMB-isolated cells stained positive for mesenchymal stromal cell markers CD90 and CD105, expressed vimentin and fibronectin and were negative for hematopoietic markers CD45 and CD34. These cells could differentiate into osteoblasts, adipocytes and chondrocytes. This study indicates that FMB may have special advantage in isolating MSC from sources such as mobilized PB, where the number of such cells is scarce.

Effectiveness of a nationwide infant immunization program against Haemophilus influenzae b. The Israeli Pediatric Bacteremia and Meningitis Group.

An ongoing nationwide prospective surveillance program for invasive H. influenzae b (Hib) disease in Israel enabled us to study the effectiveness of a national infant Hib immunization program, which included all infants born since January 1994. The vaccine used was Hib polysaccharide conjugated to outer membrane protein complex of Neisseria meningitidis b (PRP-OMPC). For the cohort born during the 3 years since January 1994, the vaccine effectiveness was 94.9% for all invasive Hib diseases and 96.6% for meningitis. The efficacy in fully immunized subjects was 98.7 and 99.5%, respectively. A herd immunity effect could be observed, since a reduction in cases also occurred among infants too young to be immunized. No increase in invasive cases caused by S. pneumoniae and N. meningitidis was observed during the study period. This is the first report outside North America and Western Europe that demonstrates a nationwide extensive reduction of invasive Hib disease within a short time of the introduction of Hib conjugate vaccines to the infant immunization program.

Early emergence of ganciclovir-resistant human cytomegalovirus strains in children with primary combined immunodeficiency.

Children with primary combined immunodeficiency (CID) and human cytomegalovirus (HCMV) infection often deteriorate despite antiviral therapy. In this study, the emergence of ganciclovir-resistant strains was examined in 6 children with CID and HCMV infection, using sequence analysis of the HCMV UL97 gene and virus susceptibility assays. Mutations in the proposed ATP binding site associated with ganciclovir resistance were found in 4 of the 6 children. In 1 patient with B severe CID, an unusual multiplicity of mutations was found in the UL97 substrate binding domain between aa 590-606. All mutations were detected within 10 days to 3 weeks from initiation of therapy. The emergence of resistant strains in children with CID appears earlier than in other groups of HCMV-infected patients. These findings may have relevance to the cellular pathways involved in viral DNA repair and mutagenesis, and they indicate the need for early and frequent genotypic monitoring and prompt therapeutic modification in this patient population.

Safety and immunogenicity of a novel mammalian cell-derived recombinant hepatitis B vaccine containing Pre-S1 and Pre-S2 antigens in children.

We tested the safety and immunogenicity of the new recombinant hepatitis B virus (HBV) vaccine produced via expression of the Pre-S1, Pre-S2 and S protein components of the hepatitis B surface antigen (HBsAg). A reduced dose (2.5 micrograms) of the vaccine (Bio-Hep-B) was tested in children aged 4-9 years and was compared to a 10 microgram dose of a licensed vaccine (Engerix-B) in a randomized manner. Both vaccines were administered in 3 doses (0, 1 and 6 month intervals). Adverse events were collected 5 days following each vaccination by a diary card provided to the parents. Immunogenicity was tested by measuring anti-hepatitis B surface antibody (anti-HBs). A total of 217 children were enrolled (162 in the Bio-Hep-B group and 55 in the Engerix-B group). Total adverse events were observed in 35% of the Bio-Hep-B group and 33% of the Engerix-B group, with no differences when each of the potential adverse events was considered. Titers were within the expected range (geometric mean titers post dose 1, 2 and 3 were 45.8, 8360.2 and 1445.7, respectively, for Bio-Hep-B and 36.3, 10316.1 and 1898.7, respectively, for Engerix-B). A trend toward better immunogenicity with Bio-Hep-B was observed at early visits when measured by both seroconversion (anti-HBs > or = 2.1 mIU ml-1) rate and seroprotection (anti-HBs > or = 10 mIU ml-1) rate; however, statistical significance was not reached. We conclude that in children, Bio-Hep-B vaccine at a reduced dose was as safe and as immunogenic as Engerix-B given at the regular pediatric dosage.

Evaluation of an enzyme immunoassay for the detection of Cryptosporidium spp. in stool specimens from infants and young children in field studies.

Diagnosis of Cryptosporidium is made by the identification of oocysts in stool specimens. Screening in field studies relies mainly on acid-fast staining followed by microscopic examination. The more sensitive immunofluorescent antibody (IFA) staining method is time-consuming, may involve technical difficulties, and is extremely costly as a screening procedure in field studies. We evaluated the diagnostic utility of a commercially available enzyme immunoassay (EIA), which detects Cryptosporidium-specific antigen, in 204 unprocessed stool specimens obtained from patients less than three years of age from a field study in southern Israel. When compared with the routine screening procedure applied in this field study (screening by acid-fast staining and microscopy after concentration, and confirmation of positive results by IFA), both the sensitivity and specificity were 98%. Of 139 specimens negative by microscopy, 13 (9.3%) were positive by the EIA. Eleven of these were confirmed by inhibition with antibody to Cryptosporidia-specific antigen. The EIA is an important tool for identifying Cryptosporidium in fecal specimens in field studies since it is sensitive, specific, simple to use, and unaffected by the presence of a preservative.

Clinical features and epidemiology of invasive Kingella kingae infections in southern Israel.

OBJECTIVE: To characterize the clinical spectrum and epidemiology of invasive Kingella kingae infections in children living in southern Israel. DESIGN: Five-year observational, descriptive study. POPULATION: Children in whom K. kingae was isolated from blood or other normally sterile body fluid. RESULTS: Twenty-five patients with invasive K. kingae infection (13 male and 12 female) were identified. Twenty-four of these children were younger than 2 years. The annual incidence was 14.3, 27.4, and 31.9 cases per 100,000 children < or = 4 years, < or = 24 months, and < or = 12 months, respectively. Seventeen (68%) of 25 patients sought treatment between July and December. Concomitant upper respiratory tract infection or stomatitis was observed in 14 (56%) of the patients, suggesting a respiratory or buccal source for the infection. Four children were bacteremic: 2 of them suffered from a lower respiratory tract infection, and the remaining 2 had bacteremia with no evident focal infection. Twenty-one children had skeletal infections and none of them was bacteremic; 16 had septic arthritis, 3 had osteomyelitis, 1 had both osteomyelitis and septic arthritis of the adjacent joint, and 1 had dactylitis of the hand. Involvement of the ankle was unusually frequent among children with septic arthritis, whereas the calcaneus was involved in 3 of the 4 children with osteomyelitis. Antibiotic treatment resulted in full recovery in all cases, and only 2 patients with septic arthritis required surgical drainage. CONCLUSION: Kingella kingae is a much more common cause of invasive infection in young children than has been previously recognized. The disease has a clear seasonal pattern, usually affects the skeletal system, frequently involves unusual bones and joints, and follows a benign course.

Human and tick spotted fever group Rickettsia isolates from Israel: a genotypic analysis.

The genomes of spotted fever group rickettsiae isolated in different geographical areas of Israel (two from ticks and four from humans, obtained over a span of 20 years) were studied by polymerase chain reaction (PCR) and restriction endonuclease fragment length polymorphism (RFLP) analysis. The human isolates were obtained from patients suffering from rickettsial disease of different degrees of severity. The PCR products obtained with five pairs of oligonucleotide primers (two primer sets derived from the 190-kDa polypeptide gene and three from the 120-kDa polypeptide gene) and cleaved with restriction endonucleases were used to study the Israeli isolates and reference Rickettsia conorii isolates. Subtle differences between the PCR-RFLP patterns of Israeli isolates and the two R. conorii reference strains (Moroccan and no. 7) were seen when the PCR products derived from the 190-kDa gene-derived primer sets were digested. All of the Israeli isolates were identical by RFLP analysis using all of the primer sets. This study showed that the Israeli spotted fever group isolates (from both ticks and humans) were genetically homogeneous by the criteria used in this study, despite the time and location differences in their original isolation, and different as a group from R. conorii.

Cryptosporidium in Bedouin and Jewish infants and children in southern Israel.

We conducted a 1-year prospective study in two clinics and a hospital in the Negev region of southern Israel to determine the epidemiological and clinical patterns of Cryptosporidium diarrhea among Jewish and Bedouin infants and children living in the same geographical area. A total of 612 episodes were studied: 398 in Bedouins and 214 in Jews, of which 449 occurred in patients with diarrhea and 164 in controls. Cryptosporidium was detected in 13 of 382 patients (3.4%) with diarrhea and in 1 of 138 controls (0.7%) (P = 0.078). In 5 of 13 Cryptosporidium-positive patients (38%) another pathogen was detected. No significant difference in Cryptosporidium detection rates was observed between Jews and Bedouins or between hospitalized or nonhospitalized patients. The frequency of Cryptosporidium detection did not differ significantly when three age-groups were compared (less than 6 months old, 7-12 and 13-36 months old). The rate of Cryptosporidium detection was similar among malnourished and well-nourished patients, as determined by weight-for-height percentiles. Cryptosporidium was detected more frequently during the summer months (8.3%) than during the rest of the year (1.2%) (P less than 0.001). Patients with Cryptosporidium diarrhea did not differ clinically from patients with other causes of diarrhea. However, they were characterized by the absence of fecal leukocytes. Cryptosporidium is not a rare cause of diarrhea in southern Israel. It is more prevalent during the hot and dry season and can be detected in a relatively high prevalence among very young infants. Its clinical features are indistinguishable from those of patients with non-cryptosporidial diarrhea.

[Articular involvement as a presenting symptom of malignancy in childhood].

The clinical and laboratory features of 6 children with arthritis or arthralgia as presenting manifestations of malignancy are described. 4 of the 6 had leukemia, 1 had a neuroblastoma and 1 had histiocytosis-X. The diagnosis of malignancy was made 2 days to 2.5 months after onset of symptoms. Although all patients presented with limp, the clinical courses suggested the need to consider leukemia and the other malignancies in the differential diagnosis.

An analysis of the characteristics of the electromyogram recorded from the mesometrium in the ovariectomized nonpregnant ewe and its similarities with and differences from the electromyogram obtained from the myometrium: the effect of estrogens.

Mesometrial and myometrial electromyograms have been recorded from ovariectomized nonpregnant ewes. After infusion with saline solution, the frequency of electromyographic events lasting less than 180 seconds in the myometrium was not different from the frequency of their occurrence in the mesometrium. During infusion of estradiol at 100 micrograms/24 hours, the frequency of events less than 180 seconds increased significantly in the myometrium but was unchanged in the mesometrium. Contracture activity (electromyographic events lasting more than 180 seconds) was significantly greater in the mesometrium than in the myometrium before estradiol administration. During estradiol infusion, contracture activity remained unchanged. These findings suggest that the mesometrial muscle may be a useful model to study the effects of various stimulators and inhibitors of uterine muscle function in the absence of stretch and local paracrine effects that occur in the uterus itself.