The impact of surgical wound bacterial colonization on the incidence of surgical site infection after lower limb vascular surgery: a prospective observational study.

OBJECTIVE: To study the relationship between surgical wound bacterial colonization and the development of surgical site infection (SSI) after lower limb vascular surgery. SSI is a major problem after lower limb vascular surgery. Most SSIs in vascular surgery are caused by Staphylococcal species that are part of normal skin flora. A prospective observational investigator blind study to examine quantitative and qualitative analysis of surgical wound bacterial colonization and the correlation with the development of SSI has been conducted. METHODS: The study cohort comprised 94 consecutive patients with 100 surgical procedures. Swabs for microbiological analyses were taken from surgical wounds at four different time intervals: before surgery, just before the surgical area had been scrubbed, at the end of surgery, and on the first and second postoperative days. Postoperative complications were recorded. RESULTS: Three hundred and eighty-seven skin bacterial samples from 100 surgical wounds were analyzed. The most common bacteria isolated were coagulase-negative staphylococci (80%), Corynebacterium species (25%), and Propionibacterium species (15%). In 13 (62%) cases, the same bacterial isolates were found in the perioperative study samples as in the infected wounds. The incidence of SSI was 21%. Multivariate analysis revealed that high bacterial load on the second postoperative day and diabetes independently increased the risk of SSI. Elective redo surgery was protective against the development of SSI. CONCLUSIONS: A high bacterial load in the postoperative surgical wound independently increases the risk of the development of SSI after lower limb vascular surgery.

IL-1 RN 2/2 genotype and simultaneous carriage of genotypes IL-1 RN 2/2 and IL-1beta-511 T/T associated with oesophagitis in Helicobacter pylori-negative patients.

BACKGROUND: Interleukin-1 receptor antagonist genotype 2/2 is associated with a prolonged and enhanced inflammatory response. It is suspected of being a risk factor for atrophic gastritis and gastric cancer and for some autoimmune diseases. No specific genetic risk factors for oesophagitis have been identified so far and there are no reports of IL-1 polymorphism in relation to oesophageal disease. METHODS: We studied the IL-1RN, IL-1beta-511 and IL-1beta + 3953 polymorphisms in an unselected series of 142 adult patients scheduled for gastrointestinal endoscopy because of dyspepsia. The control group consisted of university staff and students (n = 179). Helicobacter pylori status was determined by antibody testing and bacterial detection. RESULTS: Endoscopic oesophagitis was noted in 40 patients. The IL-1RN 2/2 genotype was significantly more prevalent in the patients with H. pylori-negative oesophagitis than in the control subjects (27% versus 9%; OR 3.574, CI 1.23-10.35, P = 0.034) or in the dyspeptic patients (27% versus 7%; OR 5.089. CI 1.51-17.11, P = 0.009). IL-1beta-511 T/T genotype tended to be more frequent in the H. pylori-negative patients with oesophagitis than in the control subjects (P = 0.071). The strongest association was between the simultaneous carriage of genotypes IL-1RN 2/2 and IL-1beta -511 T/T and H. pylori-negative oesophagitis. where the combined genotype was more prevalent than in the control subjects (23% versus 6%; OR 4.492, CI 1.40-14.46, P = 0.012) or the dyspeptic patients without oesophagitis (23% versus 3%: OR 9.706. CI 2.12-44.42, P = 0.003). CONCLUSIONS: The results indicate that the IL-1RN 2/2 genotype and the carriage of combined genotypes IL-1RN 2/2 + IL-1beta-511 T/T are associated with H. pylori-negative oesophagitis. This is the first report on the association between IL-1 gene polymorphism and oesophagitis.

HLA DQ alleles and interleukin-10 polymorphism associated with Chlamydia trachomatis-related tubal factor infertility: a case-control study.

BACKGROUND: The relationship between Chlamydia trachomatis tubal factor infertility (TFI) and the host's immunoregulatory genes was studied. METHODS: Cell-mediated immune responses to C. trachomatis and chlamydial heat shock protein (CHSP60) were determined by lymphocyte proliferation assay. HLA-DQ alleles and interleukin-10 (IL-10) promoter polymorphism (-1082 A/G) were analysed in 52 TFI cases and in 61 controls by PCR. RESULTS: HLA-DQB1 or DQA1 alleles did not significantly differ between the TFI group and the control group. However, DQA1*0102 and DQB1*0602 alleles together with IL-10 -1082AA genotype were found significantly more frequently in the TFI patients than in the controls (0.18 and 0.02 respectively; P = 0.005). Five (22%) of the 23 patients who had a positive lymphocyte proliferative response to CHSP60 were positive also for IL-10 -1082AA and for the HLA-DQA1*0102 and HLA-DQB1*0602 alleles. CONCLUSIONS: Our results reveal an association of a cellular immune response to CHSP60, HLA class II alleles and IL-10 promoter genotypes in patients with chlamydial TFI.

Effect of CD14 promoter polymorphism and H. pylori infection and its clinical outcomes on circulating CD14.

CD14 is a pattern recognition receptor on the membranes of monocytes and macrophages for several microbial products, of which lipopolysaccharide (LPS) is the best known. A shed form of CD14 is present in serum. As the CD14 gene promoter polymorphism -159C/T and some bacterial infections may affect the sCD14 levels, we compared the impact of both the CD14 promoter polymorphism and Helicobacter pylori infection on serum sCD14 levels in 201 dyspeptic patients (group 1) who had undergone gastroscopy, and 127 staff members (group 2) with no endoscopy. sCD14 was measured from the sera by a commercial enzyme immunoassay (EIA), and CD14 genotyping was carried out with PCR. Helicobacter pylori infection was detected by serology and/or culture or PCR. sCD14 levels were elevated in the subjects carrying the T allele (CT or TT genotype) in both groups when compared with subjects with the CC genotype. Overall, H. pylori-positive subjects tended to have higher sCD14 levels compared with H. pylori-negative subjects. In group 1 consisting of dyspeptic patients, those with gastric ulcer, gastric erosion or duodenal ulcer had significantly elevated levels of sCD14 compared with the patients with normal endoscopic findings or macroscopic gastritis. The recent use of NSAIDs was also associated with enhanced sCD14. Thus, we were able to show several factors, one genetic and the other environmental (H. pylori infection and mucosal lesion), to have an impact on sCD14.

Promoter region polymorphism of the CD14 gene (C-159T) is not associated with psoriasis vulgaris.

Keratinocytes of psoriatic skin show aberrant expression of membrane-bound CD14 (mCD14). In addition, soluble CD14 (sCD14) is elevated in the sera of psoriatic patients. The mechanisms leading to increased CD14 expression and secretion in psoriasis are poorly understood. A bi-allelic polymorphism in the promoter region of the CD14 gene controls CD14 expression on monocytes and sCD14 levels in the sera of healthy subjects. In this context, we explored the CD14 promoter region genotypes of 63 Finnish patients with psoriasis and 126 non-psoriatic controls using a new ARMS-PCR method. No differences in the CD14 genotype frequencies were found between the groups. Thus, our results suggest that the enhanced CD14 expression in psoriasis is not attributable to functional variants of CD14 (-159C/T).

Rapid laboratory diagnosis of ulceroglandular tularemia with polymerase chain reaction.

Tularemia is a zoonotic disease which, in Scandinavia, is usually acquired through a mosquito bite. As the infecting organism, Francisella tularensis, is highly virulent the culturing of F. tularensis has generally been avoided. PCR offers a safe way to rapidly confirm diagnosis of tularemia. The case of a 9-y-old boy with ulceroglandular tularemia is presented. The diagnosis was made rapidly with DNA amplification from a pus specimen. The efficacy of ciprofloxacin treatment of tularemia in children is also discussed.

Genotyping interleukin-10 high and low producers with single-tube bidirectional allele-specific amplification.

A simple bidirectional allele-specific PCR method is described for determining the -1082 A and G alleles in the interleukin-10 (IL-10) promoter region. This polymorphism is associated with IL-10 production capacity, and it is thus interesting to see whether different infectious and autoimmune conditions are associated with it. With our method, the A and G alleles may be studied simultaneously in a single PCR reaction, as amplification of the different alleles is performed by using 3'-mismatched and partly overlapping allele-specific upstream and downstream primers around the -1082 site. The fast and simple method described here is especially suitable for large-scale association studies.

The functionally important IL-10 promoter polymorphism (-1082G-->A) is not a major genetic regulator in recurrent spontaneous abortions.

Enhanced secretion of anti-inflammatory Th2 cytokines is a characteristic feature in normal physiological pregnancy. In recurrent spontaneous abortions (RSA), however, defective production of interleukin-10 (IL-10) and other Th2 cytokines has been shown in humans. Association studies have shown that a base exchange polymorphism (guanine-->adenine) at position -1082 of the IL-10 promoter is associated with differential IL-10 production. Since factors contributing to IL-10 production appear to be important in RSA, we studied the IL-10 genotypes of 38 Finnish women with a history of three or more consecutive abortions and 131 ethnically matched healthy controls. No significant differences in the -1082 allele or genotype frequencies were found between the controls and the RSA women. The present study suggests that the IL-10 -1082 (G-->A) polymorphism is not a major genetic regulator in RSA.

cagA, vacA and iceA virulence genes of Helicobacter pylori isolates of children in Finland.

cagA, vacA s and m genotypes and iceA alleles were analyzed from Helicobacter pylori strains isolated from 17 Finnish children and 32 children of non-Finnish origin living in Finland. Twelve children in the latter group were eastern European and 15 were of African origin. Only three children of non-Finnish origin were born in Finland. The vacA sla subtype was more prevalent in the isolates from Finnish children than African children (76% vs. 7%, P<0.001); vacA s1b frequencies were 5% and 67%, respectively (P<0.001). The iceA1 allele was significantly more prevalent in African than Finnish isolates (93% vs. 35%, P< 0.01). Considerable variation was noted in the frequency of vacA s1 subtypes and iceA alleles in children originating from different geographic regions, but the geographic variation of s1 subtypes resembled that described in other reports.

In vitro antibiotic susceptibility of Francisella tularensis isolated from humans and animals.

The in vitro susceptibility of 38 strains of Francisella tularensis (biovar F. tularensis palaearctica) was determined using Etests on cysteine heart agar plates with 2% haemoglobin. All strains were susceptible to the antibiotics traditionally used to treat tularaemia, such as streptomycin (MIC(90) 4.0 mg/L), tetracycline (MIC(90) 0.38 mg/L) and chloramphenicol (MIC(90) 0.38 mg/L), and to aminoglycosides, such as tobramycin (MIC(90) 1.5 mg/L) and gentamicin (MIC(90) 1.0 mg/L). The quinolones examined had low MIC(90)s: ciprofloxacin, 0.016 mg/L; levofloxacin, 0.016 mg/L; grepafloxacin, 0.047 mg/L; and trovafloxacin, 0.032 mg/L. In contrast, all the strains were resistant to beta-lactams and azithromycin. Quinolones thus seem to be promising drugs for the treatment of tularaemia.

HLA-DQA1 alleles and the presence of Helicobacter pylori antibodies.

Recent reports suggest that the HLA-DQA1 gene may be important in determining susceptibility to and outcome of Helicobacter pylori infection. To determine if there is an association between HLA-DQA1 alleles and H. pylori antibodies, DQA1 alleles and H. pylori-specific antibodies were determined in 199 random subjects of Finnish origin (mean age 43 years, range 22-69 years). H. pylori-specific class IgG antibodies were measured using the EIA method (Pyloriset-EIA-G, Orion Diagnostica, Espoo, Finland). HLA-DQA1 typing was carried out using PCR-SSP (PCR with sequence-specific primers). There were 64 subjects with H. pylori-specific class IgG antibodies (ab+) and 135 subjects without H. pylori-specific class IgG antibodies (ab-). Gene and phenotype frequencies of HLA-DQA1 alleles were similar in the ab+ and ab- subjects (P = NS). The data suggest that no single HLA-DQA1 allele is associated with the presence of serum antibodies against H. pylori.

HLA-G polymorphism in Finnish couples with recurrent spontaneous miscarriage.

HLA-G is a class I major histocompatibility complex gene that is expressed in cytotrophoblasts at the materno-fetal interface. It has been suggested that HLA-G could play a key role in materno-fetal immunological interactions during pregnancy. To investigate whether there is an association between HLA-G locus and recurrent spontaneous miscarriage, HLA-G alleles were determined by a PCR-RFLP method in 38 couples with recurrent spontaneous miscarriage and in 26 random control couples. In this series parental HLA-G sharing, extended HLA-G/A haplotypes and the frequencies of the HLA-G alleles were similar in the two groups. Thus, our data suggest that there is no detectable relation between susceptibility to recurrent spontaneous miscarriage and HLA-G locus.

HLA-G polymorphism and allelic association with HLA-A in a Finnish population.

The polymorphism of the HLA-G gene can be identified by PCR-RFLP analysis. This short communication describes the PCR-RFLP analysis of HLA-G polymorphisms in exons 2 and 3 and the association of different HLA-G and HLA-A alleles in 26 healthy Finnish families.