Head and neck squamous cell carcinomas (HNSCC) are a highly heterogenous disease which can be induced by two main carcinogens - tobacco and/or alcohol, or by HR HPV infection. This work examined 60 paraffin-embedded biopsies of head and neck carcinomas after histological verification. HPV infection, including its specific types in various HNSCC areas, was studied using multiplex qPCR. Expression levels of p16INK4A and p53 were detected by subsequent IHC analysis as being potential diagnostic markers. Based on the assumption that patients with HNSCC could benefit from anti-EGFR therapy (cetuximab), but the predictors are not yet defined, analyses of point mutations of ras genes (Kras, Nras) were carried out using multiplex qPCR and sequence analysis of the Braf gene. All statistical data were processed by Chí-x2 test.HPV infection was detected in 23.34 % of cases with HNSCC, of which 100 % were HPV 16, which is the most frequently infection found in the oropharyngeal region. Using IHC analysis, a positive expression of P16INK4A was detected in 100 % of HPV-positive HNSCC while this expression was discovered to be highly correlated with HPV infection. Furthermore, a correlation between p53 and HPV-negative HNSCC was proved. The mutation incidence was the highest in the Kras gene (codon 12 and codon 146), Nras (codon 12) and Braf. A correlation between tumor location in the oropharyngeal region and Kras mutations was proved. The HPV infection correlated with Kras mutations in case of codon 146 but on the grounds of low amount of output data, these figures could be irrelevant. In one case, c.1808 G>A, protein 603 Arg>Gln mutation was found in the Braf gene but its correlation with head and neck carcinomas has not been described yet (Tab. 2, Fig. 2, Ref. 24). Keywords: head and neck carcinomas, biopsy, HPV types, PCR, p16INK4A, p53, molecular predictors, Kras, Nras, Braf.

GOAL:To find out a serology prevalence of Helicobacter pylori (Hp) infection in patients suffering from rheumatoid arthritis (RA) and to investigate its relationship to pharmacotherapy modes.METHOD:To determine Hp IgG class and IgA class antibodies by ELISA method in hospitalised patients with active RA according to ACR criteria (1987).RESULTS:137 patients with RA were examined--20 men, 117 women, an average age was 54.6 +/- 13.1, an average length of RA in these patients was 12.46 +/- 9.75, positive RF/LFT had 88% of patients. Hp ELISA IgG antibodies were confirmed in 57% (78/137) of patients with RA, Hp ELISA IgA antibodies were confirmed in 60% of examined patients (82/137), and IgG + IgA antibodies simultaneously were found in 51% (70/137) of examined patients. A comparison of basic demographic data, specific parameters of the disease, and clinical signs has not confirmed any differences between groups of Hp positive and Hp negative patients. Patients with both types of Hp antibodies (IgG + IgA) had more frequent signs of RA in other places than joints (p = 0.046). RA patients with anti-Hp IgG antibodies more frequently used anticoagulation drugs (p = 0.029) while patients with anti-Hp IgA antibodies more frequently used methotrexate (p = 0.01).CONCLUSIONS:Results point out more frequent incidence of Hp IgA antibodies in RA patients treated with methotrexate and more frequent incidence of both IgG and IgA antibodies in patients with RA signs in other places than joints.

PURPOSE:To report retrobulbar neuritis caused by Borrelia afzelii culturally proved from cerebrospinal fluid (CSF).METHODS:A 23 year old female underwent ophthalmologic, laboratory and other auxilliary examinations.RESULTS:CSF cultures grew spirochetal microorganisms, serotyped by monoclonal antibodies as Borrelia afzelii. Following the serological and cultural results, treatment with doxycycline 200 mg daily was started and kept for three weeks. Gradual improvement of the visual acuity of the right eye was observed with full recovery to 20/20.CONCLUSIONS:Borrelia infection should be considered in the differential diagnosis of retrobulbar neuritis. CSF should be examined also culturally. (Ref. 5.)

Over a 10-y period (1989-99) we prospectively evaluated all patients with fungaemia among 16,555 admissions (21,004 blood cultures) at a national cancer referral institution in the Slovak Republic. A prospective protocol was completed on 140 patients with fungaemia, which was then analysed in terms of aetiology, clinical characteristics, potential risk factors and outcome. The most frequently isolated organism was C. albicans, in 75 patients (52.9%), followed by non-albicans Candida spp. in 45 patients (32.1%). Non-Candida spp. yeasts represented 16 episodes in 16 patients (11.4%). Moulds caused 4 episodes in 4 patients (3.6% of all fungaemias) and all were caused by Fusarium spp. Mucositis (p = 0.025), > or = 3 positive blood cultures (p = 0.02), acute leukaemia (p = 0.00001), neutropenia (p = 0.0015), quinolone prophylaxis (p < 0.000005) and breakthrough fungaemia (p = 0.004) during prophylaxis with fluconazole (p = 0.03) and itraconazole (p = 0.005) were significantly more associated with non-Candida than C. albicans spp. Furthermore, attributable mortality was higher in the subgroup of non-Candida than C. albicans spp. (50.0 vs. 18.7%, p < 0.02). The only independent risk factor for inferior outcome was antifungal therapy of < 10 d duration (odds ratio 2.1, 95% confidence interval, p < 0.001). Aetiology, neutropenia and mucositis were not independent risk factors for higher mortality in multivariate analysis; however, they were risk factors for inferior outcome in univariate analysis (p < 0.05-0.005).

The aim of this multicenter survey was to assess risk factors and mortality in patients with persistent fungemia (PF). Cases of persistent fungemia, defined as positive blood culture for at least 3 causative days of antifungal therapy were selected. Forty cases of persistent fungemia (lasting more than 3 days) were compared with 270 non-persistent fungemias appearing within the same period, and analyzed by univariate and multivariate analysis for risk factors and outcome. The median number of days of positive culture was 4.4 (3 - 20): 22 episodes were due to Candida albicans, 1 due to non-albicans Candida spp., 6 episodes due to non-Candida spp. Yeasts: 15 were catheter related, 16 patients had yeast-infected surgical wounds, 12 were neutropenic, 4 cases were caused by species resistant in vitro, 2 to amphotericin B (Trichosporon spp.) and 2 to fluconazole (C. laurentii, C. glabrata). Fifteen patients (37.5%) died, 7 of whom due to fungemia. Nineteen cases had one known risk factor (10 had infected wound, 4 infected vascular catheter, 3 were neutropenic and 2 had inappropriate therapy). Fourteen cases had two known risk factors (4 had wound and infected catheter, 4 neutropenia and infected catheter, 2 neutropenia and resistant organism, 4 other combinations. Two cases had 3 known risk factors and one had 4 risk factors for persistent fungemia. Artificial ventilation, C. glabrata etiology, non-Candida spp. yeasts such as Trichosporon spp. and Cryptococcus spp. and prior surgery were significantly associated with persistent fungemia in univariate, whereas only C. glabrata etiology in multivariate analysis. Breakthrough fungemia during empiric therapy with fluconazole was also observed more frequently in patients with persistent fungemia. However, there was no difference in both attributable and overall mortality between both groups.

The aim of the present study was to determine the prevalence of Helicobacter pylori infection in a group of patients hospitalized at the clinic of rheumatology who presented peptic ulcers and erosions associated with nonsteroidal antiinflammatory drugs (NSAIDs). Of a group of 4,256 hospitalized patients receiving therapy with NSAIDs, 221 patients with persistent dyspepsia underwent endoscopic examination of the upper segment of the gastrointestinal tract. Among them, mucosal abnormalities in the stomach and duodenum were confirmed in 69 patients. H. pylori was found in 42% (29/69) of the examined patients. Peptic ulcers were confirmed in 19 patients. Localization was gastric in 12 patients and duodenal in seven. H. pylori was found in only 17% of the patients (2/12) with gastric ulcers, but in as many as 86% (6/7) of those with duodenal ulcers. Patients with H. pylori taking NSAIDs were at higher risk of developing duodenal mucosal abnormalities, both ulcers (OR: 10.17; 95% CI: 1.08-23.88, p = 0.013) and erosions (OR: 2.67; 95 CI: 1.94-3.66, p = 0.001). Concomitant administration of corticoids and NSAIDs did not increase the risk of gastrotoxicity in patients with positive finding of H. pylori (OR: 0.32; 95% CI: 0.1-0.96). In conclusion, a close association was found between H. pylori infection and duodenal ulcers and erosions, but not between gastric ulcers and gastric erosions in a group of patients hospitalized for rheumatic diseases and undergoing NSAID therapy.

The objective of this study was to assess risk factors and the outcome of breakthrough fungaemias (BFs) occurring during fluconazole (FLU) therapy in non-cancer and non-HIV individuals. Thirty-three fungaemias occurring during therapy with FLU among a total of 310 fungaemias observed within a 10-y national survey were analysed. The agar disk diffusion method was used for antifungal susceptibility testing and the Vitek system for species identification. Univariate and multivariate analysis was performed to determine risk factors for BF. All BFs were due to species known to be susceptible to FLU: Candida albicans (25/33), C. parapsilosis (6/33) and C. guillermondii (2/33). The mean number of positive blood cultures per episode was 2.4. The MIC of Candida spp. to FLU was 0.5-8 mg/ml (all strains were susceptible in vitro). Neonatal age (< 4 weeks), very low birth weight, prior surgery, central venous catheter placement, artificial ventilation, total parenteral nutrition and C. parapsilosis were significantly related to BF in univariate analysis, but only central venous catheter placement was significantly related in multivariate analysis. However, the outcome of BFs and non-BFs was similar. All BFs occurred in non-HIV patients who were not previously treated with azoles, and were caused by in vitro FLU-susceptible species (C. albicans and C. tropicalis). Thus factors other than in vitro susceptibility play a role in BFs.

Breakthrough fungaemias due to Candida albicans and Candida parapsilosis appearing during fluconazole therapy in neonates and infants were assessed for risk factors and outcome. Forty fungaemias occurred during therapy with fluconazole within a 12 year national survey and were compared with 161 cases of non-breakthrough paediatric fungaemias. The agar disc diffusion test method was used for antifungal susceptibility testing and the Vitek system for species identification. Univariate and multivariate analysis for risk factors for breakthrough fungaemia were carried out. All the fungaemias were a result of strains susceptible to fluconazole at 0.25-4 mg/L in vitro [C. albicans (85%) and C. parapsilosis (15%)]. The mean number of positive blood cultures per episode was 2.2. Sixteen children had 'early' breakthrough fungaemias (within 4-5 days) and 24 fungaemias appeared on day 6 and later. Mean fluconazole MICs in the 'early' group were 1.2, and 2.8 mg/L in the 'late' group (P < 0.03, t-test). However, no difference was observed in the average dose of fluconazole used in the two groups. Neonatal age, total parenteral nutrition, very low birth weight, before surgery, central or umbilical venous catheterization and artificial ventilation were all significantly related to breakthrough fungaemia in univariate analysis but only central or umbilical venous catheterization were significant in multivariate analysis. The outcome of breakthrough fungaemia was better overall and attributable mortalities in non-breakthrough fungaemia was significantly higher in comparison with breakthrough fungaemia.

There are few published articles on fungaemia in the elderly, although geriatric patients are known to have different clinical courses and worse prognoses with infections [1,2]. Here, we present the aetiology, risk factors and outcome of candidaemia in 20 patients, who were 70 or more years old. All fungaemias occurred in four university hospitals in Bratislava and Trnava within the last 10 years. Detailed characteristics of 20 cases (median age 72.3 years) are in Table 1. Eight of the 20 had cancer, 10 diabetes and 14 abdominal or thoracic surgery as an underlying disease and 18 of 20 were pre-treated with broad spectrum antibiotics. Sixteen had a central venous catheter inserted and 12 stayed on the ICU more than 5 days. Candida albicans caused fungaemia in 12 and non-albicans Candida spp. in eight patients (four C. parapsilosis and four C. glabrata), and five were infected with Candida spp. resistant to at least one of the antifungals tested. Fluconazole was used in 10 and amphotericin B in two cases. Eight patients were not treated of whom six died. These were mainly patients who had undergone abdominal surgery because many surgeons did not consider the isolation of Candida spp. as an indication for systemic antifungal therapy. The overall mortality was 55%; 30% of elderly patients died of infection (attributable mortality) and 25% with underlying disease accompanying fungaemia. Table 2 gives univariate analysis 2 test with Mantel– Haenzsel modification and Yates correction) for 20 fungaemias in elderly patients and 290 fungaemias collected in a national survey within the same period in non-elderly patients (adults in age 18–69). Of the risk factors, previous surgery (70.0% vs. 21.0%, P 0.001), C. tropicalis (20.0% vs. 3.4%, P 0.01), C. glabrata (20.0% vs. 2.0%, P 0.002) and absence of antifungal therapy (40.0% vs. 17.3%, P 0.02) were significantly associated with fungaemia in elderly patients compared with the other patient population. Only 45% of elderly patients survived (55% overall mortality) and 30% died from fungaemia (attributable mortality). It is difficult to compare our results with other studies as we are not aware of any report on a similar group of this age. There have been four large studies on fungaemia in last 5 years, three in US [3–5] and one in Europe [6]. The proportion of patients 65 years was only given in two studies [3,4] and were 5% and 6% respectively. However, no differences in specific risk factors, aetiology or mortality were given in the subgroup of elderly patients in those studies. As seen from our results, appropriate and early antifungal therapy is under-considered in the elderly. Only 60% of fungaemias in patients over 70 were appropriately treated and the absence of appropriate therapy was associated with a significantly inferior outcome (55.0% vs. 39.0% mortality). Special attention should be given to elderly patients, especially those with diabetes and after surgery, who have positive blood cultures for Candida spp. or who do not respond to broad spectrum antibiotic therapy and have central venous catheter inserted. Early therapy with flucytosine (which is less nephroand hepato-toxic than amphotericin B) should be administered and the vascular catheter (if suspected as source of fungaemia) immediately changed. A part of this study was presented at the IDSA Annual Meeting, Denver, 8–11 November 1998 (Abstract no. 99a). * Corresponding author. Tel.: +421-7-324308. E-mail address: krcmery@spamba.sk (V. Krcmery).

The consumption of antimicrobial agents in a Slovakian national cancer institute from 1989-1996 was compared with resistance rates in clinically significant blood culture isolates. We observed an increase in resistance of viridans streptococci to penicillin and of enterococci to ampicillin. Resistance to vancomycin and teicoplanin was stable over the entire period despite a 20-fold increase in vancomycin consumption. Nor did we observe increased resistance to ofloxacin despite a 10-fold increase in consumption. Consumption of aminoglycosides and resistance levels were both stable. A different situation was observed with third-generation cephalosporins, where resistance of Pseudomonas aeruginosa, Stenotrophomonas maltophilia and Acinetobacter spp. to ceftazidime and cefotaxime increased with increasing consumption. Resistance of Enterobacteriaceae to cefotaxime and ceftazidime was stable. Resistance to imipenem did not change significantly. However, the number of Stenotrophomonas maltophilia bacteremias increased significantly after imipenem was introduced in 1991. Because of improved outcome in bacteremia, an increased incidence of both gram-negative and gram-positive bacteremia led to only a slight increase in associated mortality.

At the beginning of new millennium, Lyme borreliosis is still the subject of intensive research, polemic discussions and open questions. The authors present minute analysis of the issues associated with Lyme borreliosis, concentrating on biological aspects and taxonomic classification of the agent, co-transmission and co-infection, diagnostic criteria and their validity, laboratory diagnostics an therapy of the disease including perspectives of active immunisation in the future. Special attention is paid to open questions in clinical and laboratory diagnostics of the disease and to the prospects for the near future. The authors also discuss the importance of international and interdisciplinary cooperation in the process of articulation of diagnostic criteria and recommended procedures for laboratory diagnostics. (Ref. 17.)

The aim of this prospective study on fungemia in children with cancer compared with adults with cancer appearing during the last 10 years in a pediatric hospital and in national cancer institutions was to investigate risk factors, etiology, therapy, complications and outcome. Univariate analysis showed significant differences in 35 children with cancer and fungemia in comparison with 130 cases of fungemias in adults with cancer. It was found that (1) therapy with corticosteroids (40 vs 18.5%, P <0.03), (2) breakthrough fungemia during ketoconazole prophylaxis (20 vs 7.7%, P <0.025), and (3) meningitis as a complication of fungemia (11.4 vs 0.8%, P <0.001) occurred more frequently in the pediatric subgroup with fungemia. Candida albicans was more common as the causative agent of fungemia among adults (58.5 vs 37.1, P <0.02) than in children. However, mortality was similar in children with cancer and in adults with cancer and fungemia (31.4 vs 23.1%, NS). Comparison of risk factors revealed no differences between adults and children with cancer and fungemia except in etiology, breakthrough fungemia during prophylaxis with ketoconazole, prior therapy with corticosteroids and meningitis as a complication. The outcome was also similar in pediatric and adult cancer patients with fungal bloodstream infection.

Nosocomial meningitis is an infrequent complication of craniocerebral trauma, ventriculoperitoneal shunt insertion or various neurosurgical procedures in intensive care of premature neonates 1. I ts incidence in Slovak University Hospitals is low, approximately l/2500 admissions/year. The commonest pathogens are staphylococci, followed by Enterobacteriaceae, Pseudomonas aeruginosa, Acinetobacter spp., Group B streptococci and Candida spp. However, the proportion of enterococci is increasing and represents the fifthsixth most common pathogen with 1-5% among those pathogens causing nosocomial meningitis (NM) 2. Reports of enterococcal meningitis are rare 3,4. Here we present 10 cases of enterococcal NM from a national survey of NM within the last 5 years from 4 major pediatric University hospitals in the Slovak Republic (5.5 mil population). Table 1 contains a description of l0 patients (27 days-16 years old). All meningitis cases were caused by Enterococcus faecalis and all were sensitive to vancomycin, gentamicin and all but one also to ampicillin and chloramphenicol. Six patients had prior neurosurgery, two hydrocephalus after previous meningitis, two craniocerebral trauma, 2 were premature neonates and 2 had perinatal pathology. In 4, a ventriculoperitoneal shunt (VP) was reinserted because of shunt infection. All 10 had central venous catheter (CVC), in 4 NM was accompaJournal of Chemotherapy Vol. 11 n. 4 (109-111) 1999