Use of complementary markers in assessing glycaemic control in people with diabetic kidney disease undergoing iron or erythropoietin treatment.

AIMS: HbA(1c) values are unreliable in patients with diabetes who have chronic kidney disease who receive iron and/or erythropoiesis stimulating agents. The study aimed to evaluate the utility of the complementary glycaemic markers glycated albumin, fructosamine and 1,5 anhydroglucitol in this group of patients. METHODS: A prospective study of patients with Type 2 diabetes and chronic kidney disease stage IIIB/IV undergoing intravenous iron or erythropoiesis-stimulating agent therapy. Glycaemic control was monitored using HbA(1c), seven-point daily glucose thrice weekly, continuous glucose monitoring, glycated albumin, fructosamine and 1,5 anhydroglucitol. RESULTS: Fifteen patients [9 men; median age 72 years (interquartile range 68-74), follow-up period (16.4 ± 3.7 weeks)] received parenteral iron; 15 patients [11 men; 70 years (interquartile range 62-75), (17.3 ± 3.3 weeks)] received erythropoiesis-stimulating agent. HbA(1c) fell following treatment with both iron [57 mmol/mol (7.4%) to 53 mmol/mol (7.0%), P < 0.001] and erythropoiesis-stimulating agent [56 mmol/mol (7.3%) to 49 mmol/mol (6.6%), P = 0.01] despite mean blood glucose remaining unchanged (iron: 9.55 to 9.71 mmol/l, P = 0.07; erythropoiesis-stimulating agent: 8.72 to 8.78 mmol/l, P = 0.89). Unlike HbA1c , the glycated albumin, fructosamine and 1,5 anhydroglucitol levels did not change following iron [glycated albumin (16.8 to 16.3%, P = 0.10); fructosamine (259.5 to 256 µmol/l, P = 0.89); 1,5 anhydroglucitol (54.2 to 50.9 µmol/l, P = 0.89)] or erythropoiesis-stimulating agent [glycated albumin (17.9 to 17.5%, P = 0.29), fructosamine (324.3 to 306.0 µmol/l, P = 0.52), 1,5 anhydroglucitol (58.2 to 46.7 µmol/l, P = 0.35)]. Despite this, HbA(1c) was consistently the marker most closely related to mean blood glucose before and after each treatment (R range 0.7-0.88). CONCLUSIONS: These data indicate that HbA(1c) was statistically most closely related to mean blood glucose, but clinical trends in glycaemia in patients undergoing iron or erythropoiesis-stimulating agent therapy are likely best assessed by including one of these additional glycaemic markers.

Phenotypic and genetic characterisation of methicillin-resistant Staphylococcus aureus strains isolated from the university hospitals of Debrecen.

The purpose of this study was to characterise methicillin-resistant Staphylococcus aureus (MRSA) strains isolated in 2005 at the university hospitals of Debrecen, Hungary. Three hundred and thirty-nine MRSA strains were isolated from 102 patients at 18 different clinics. Their sensitivity to oxacillin and ten other antibiotics was determined. For genotypic analysis, phage typing and pulsed-field gel electrophoresis (PFGE) was performed. The rate of MRSA strains increased to 7.2% in 2005, especially at the clinics of surgery, pulmonology and paediatrics. No vancomycin- or teicoplanin-resistant strains were found. The resistance to erythromycin, clindamycin and ciprofloxacin was nearly 100% and multi-resistance was very frequent. Fifty-eight percent of the isolates belonged to mixed phage types and 8% was non-typable. One PFGE clone contained 58.2% of all strains and two further major clones were found at a separately located clinical block, indicating intra-hospital spread. We can conclude that MRSA exhibits an increasing nosocomial problem also in Hungary.

Survivin promoter polymorphism and cervical carcinogenesis.

BACKGROUND: Survivin, a novel member of the inhibitor of apoptosis family, plays an important role in cell cycle regulation. A common polymorphism at the survivin gene promoter (G/C at position 31) was shown to be correlated with survivin gene expression in cancer cell lines. AIM: To investigate whether this polymorphism could be involved in the development of human papillomavirus (HPV)-associated cervical carcinoma. METHODS: Survivin promoter polymorphism was detected in patients with cervical cancer, in patients with equivocal cytological atypia and in a control population using polymerase chain reaction (PCR-restriction fragment length polymorphism (RFLP) and PCR-single strand conformation polymorphism analysis. HPV was typed in patients with cervical cancer and cytological atypia using PCR-RFLP. RESULTS: No statistically significant differences were found in the genotype distributions of the survivin promoter variants among our study groups. CONCLUSIONS: The survivin promoter polymorphism at position 31 may not represent an increased risk for the development of cervical cancer, at least in the population studied here.

Immunity to oncogenic human papillomaviruses.

UNLABELLED: The establishment of human papillomavirus (HPV) infection as a major cause of several human cancer forms, notably cervical cancer, has spurred development of prophylactic and/or therapeutic HPV vaccines for prevention of cervical neoplasia. Knowledge of the immunity to HPV forms the basis for such endeavors. METHOD: A literature review of humoral and cellular immunity to HPV. The overview on human leukocyte antigen (HLA) and cervical cancer was expanded to a formal metaanalysis, where relevant articles were located by Medline search and citation analysis and graded by preassigned quality criteria on study design. RESULTS: The antibody response to the HPV particle is dominated by a neutralizing antibody response to a typespecific, conformationally dependent immunodominant epitope. Vaccines based on viral particles lacking the viral genome (virus-like particles, VLPs) have been highly successful in preventing and treating HPV infection in several animal model systems. In humans, the serum antibody response to VLPs is stable over time, also after the HPV infection has been cleared, resulting in HPV serology being used as a marker of cumulative HPV exposure in spite of the fact that a significant proportion of HPV-exposed subjects fail to seroconvert. More than 90% of HPV infections will clear spontaneously. The factors that determine whether an HPV infection is cleared or persists and increases the risk for cancer are not known, but cellular immunity is implicated. Several HLA class II haplotypes are associated with cervical cancer: DQw3 increases and DR13 decreases the risk for cervical cancer in general (odds ratios (OR) and 95% confidence intervals (CI): 1.25(1.15-1.37) and 0.69 (0.56-0.85), respectively); DR15 increases the risk for HPV16-carrying cancer (OR: 1.47; CI: 1.20-1.81); and DR7 may be either protective or increase the risk. Most cervical cancers have downregulated the expression of at least one HLA class I antigen, whereas class II expression is increased in infected epithelium. A Th2 cytokine profile is associated with progression to cervical cancer. HPV-antigen-specific proliferative responses have been detected in many studies, although it is not entirely clear whether these responses are HPV type specific or may be cross-reactive between HPV types. Specific cytotoxic T lymphocyte (CTL) responses were originally reported in only a minority of infected subjects, typically cancer patients, but with advancing technology, specific CTLs can be stimulated from about half of the women with HPV-carrying disease. In animal model systems, CTL responses can mediate clearance. CONCLUSION: The antibody response to HPV is a mediator of type-specific protective immunity, which forms the basis for prophylactic vaccine candidates. The cellular immunity to HPV is implicated as an important factor in cervical carcinogenesis, but the main targets and types of responses that mediate HPV clearance are not established.

High co-prevalence of genogroup 1 TT virus and human papillomavirus is associated with poor clinical outcome of laryngeal carcinoma.

BACKGROUND: The aetiology and factors leading to the progression of laryngeal cancer are still unclear. Although human papillomavirus (HPV) has been suggested to play a role, reports concerning the effect of HPV infection on tumour development are controversial. Recently, transfusion transmitted virus (TTV) was suggested to play a role in certain infections as a causative or coinfecting agent. AIMS: To investigate whether the development and progression of laryngeal squamous cell carcinoma is associated with coinfection with TTV and HPV. METHODS: The prevalence of TTV and HPV was investigated using the polymerase chain reaction in tissue samples from 40 healthy individuals, 10 patients with recurrent papillomatosis, five patients with papillomatosis with malignant transformation, and 25 patients with laryngeal carcinoma. The obtained prevalence data were compared and analysed statistically. RESULTS: In the 11 patients with carcinoma who had metastasis or relapse there was a high rate of coinfection with genogroup 1 TTV and HPV (eight of 11), whereas in the 14 without tumour progression no coinfection was found. Coinfection was associated with significantly lower tumour free survival in patients with carcinoma (p < 0.001). Furthermore, four of five patients who had papillomatosis with malignant transformation were coinfected with genogroup 1 TTV and HPV. CONCLUSIONS: Although the nature of cooperation between HPV and TTV needs to be investigated further, coinfection with genogroup 1 TTV and HPV appears to be associated with poor clinical outcome in laryngeal cancer.

Induction of HIV-1 replication in latently infected syncytiotrophoblast cells by contact with placental macrophages: role of interleukin-6 and tumor necrosis factor-alpha.

The syncytiotrophoblast (ST) layer of the human placenta has an important role in limiting transplacental viral spread from mother to fetus. Although certain strains of human immunodeficiency virus type 1 (HIV-1) may enter ST cells, the trophoblast does not exhibit permissiveness for HIV-1. The present study tested the possibility that placental macrophages might induce replication of HIV-1 carried in ST cells and, further, that infected ST cells would be capable of transmitting virus into neighboring macrophages. For this purpose, we investigated HIV-1 replication in ST cells grown alone or cocultured with uninfected placental macrophages. The macrophage-tropic Ba-L strain of HIV-1, capable of entering ST cells, was used throughout our studies. We demonstrated that interactions between ST cells and macrophages activated HIV-1 from latency and induced its replication in ST cells. After having become permissive for viral replication, ST cells delivered HIV-1 to the cocultured macrophages, as evidenced by detection of virus-specific antigens in these cells. The stimulatory effect of coculture on HIV-1 gene expression in ST cells was mediated by marked tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) release from macrophages, an effect caused by contact between the different placental cells. Results of this study suggest an interactive role for the ST layer and placental macrophages in the dissemination of HIV-1 among placental tissue. Data reported here may also explain why macrophage-tropic HIV-1 strains are transmitted preferentially during pregnancy.

Transcriptional activity of human papillomavirus type 16 variants having deletions in the long control region.

Transcription of the E6 and E7 viral oncogenes of human papillomavirus (HPV) type 16 is regulated by the P97 major early promoter, and enhancer and silencer elements found in the long control region (LCR). In this study, we tested the transcriptional activity of natural HPV 16 variants having long deletions in the LCR. The HPV 16 LCR regions were amplified from invasive cervical cancer specimens, and cloned into the reporter vector pALuc. Transcriptional activity of the different clones was measured by luciferase test after transient transfection into HeLa cells. The deletions found in the LCR encompassed parts of the enhancer and either the YY1-specific silencer alone or together with the CDP-specific silencer. The transcriptional activity of these deletion variants were usually reduced compared with that of the corresponding full-length clones. However, a deletion variant lacking the whole enhancer and both silencer regions retained substantial enhancer activity on the P97 promoter. These results point to the existence of a novel context-dependent enhancer element in the 5' LCR of HPV 16.

Frequent methylation of p16INK4A and p14ARF genes implicated in the evolution of chronic myeloid leukaemia from its chronic to accelerated phase.

The frequency and mechanism of p16(INK4A) and p14(ARF) gene alterations were studied in cell samples from 30 patients with Philadelphia (Ph) chromosome-positive chronic myeloid leukaemia (CML), both at diagnosis and at the onset of the accelerated phase (AP) of the disease. No alterations in the p16(INK4A) or p14(ARF) genes were found in any of the chronic phase (CP) samples. DNA sequencing analyses detected p16(INK4A) or p14(ARF) mutations in 17 AP samples. All mutations were heterozygous without loss of the other allele. Aberrant methylation of the p16(INK4A) or p14(ARF) promoters was found in 14 of 30 AP samples. The most common situation was the simultaneous methylation of both promoters. Our data indicate that p16(INK4A) and p14(ARF) are primary targets for inactivation by promoter methylation in the acceleration of CML. Transcriptional silencing of the p16(INK4A) and p14(ARF) genes may be important in the conversion of CML from the CP to the AP.

Enzyme-linked immunosorbent assay for measurement of IgG antibody to Molluscum contagiosum virus and investigation of the serological relationship of the molecular types.

An ELISA was developed for measuring levels of IgG antibodies to Molluscum contagiosum virus (MCV) using DNA typed purified virus as antigen. The assay was found to be specific and sensitive for antibodies to MCV as well as being economical in its use of antigen. A close relationship was found between antibody levels to the MCV molecular types 1, 1v and 2 by cross-testing sera on plates coated with the different molecular types of the virus as antigen (P < 0.001).

HHV-8 ELISA based on a one-step affinity capture of biotinylated K8.1 antigen.

The immunogenic envelope antigen gp35-37 of human herpesvirus-8 (HHV-8) is encoded by orfK8.1. An ELISA is described using streptavidin capture of bacterially expressed and biotinylated recombinant K8.1 antigen. The antigen capture strategy provides a simple and reliable method, which does not require high yield production and purification of the recombinant antigen before the serological assay. The specificity and sensitivity of the K8.1 ELISA were validated by gp35-37 envelope antigen Western blot and anti-lytic membrane immunofluorescence assay using lytically induced HHV-8 infected BCBL-1 cells. Under the established ELISA conditions, eight of the 10 Kaposi's sarcoma patients and five of the 180 healthy blood donors had IgG antibodies to K8.1 envelope antigen.

Frequency of p53 codon 72 genotypes in human papillomavirus associated squamous intraepithelial lesions and cervical cancer.

BACKGROUND: A recent study suggested that the p53Arg (at residue 72) homozygous genotype could be a potential genetic risk factor for cervical cancer among white women. To confirm this result we examined the proportion of p53 genotypes in a larger number of patients with cervical cancer and in patients with squamous intraepithelial lesions (SIL) compared to a control population. MATERIALS AND METHODS: We used allele-specific primers to amplify the p53Arg and p53Pro sequences and we examined the proportion of genotypes in the study populations using chi 2-test. RESULTS: The distributions of p53Arg homozygous, heterozygous and p53Pro homozygous genotypes were 63%, 27% and 10% in cervical cancer patients, 53%, 36% and 8% in individuals with SIL, and 60%, 36% and 4% in control population. Using chi-square test, no significant difference was found between genotype frequencies in the study groups. CONCLUSION: Thus, the p53Arg homozygous genotype does not seem to increase the risk of cervical cancer in Hungarian women.

Integration status of virus DNA and p53 codon 72 polymorphism in human papillomavirus type 16 positive cervical cancers.

BACKGROUND: The aim of this study was to determine the physical state of HPV16 DNA and to reveal any association between the physical state of virus DNA and pathologic or prognostic factors in HPV16 positive cervical cancers. The other aim was to estimate the role of p53 codon 72 polymorphism in disease progression. MATERIALS AND METHODS: The presence and physical state of HPV16 DNA was analysed by Southern blot hybridisation and E1-E2 specific PCRs in the primary tumours and pelvic lymph nodes of 85 cervical carcinoma patients. Results Integrated HPV16 DNA was found in 32 out of 41 (78%) primary tumours and 2 out of 22 (95%) lymph nodes carrying HPV16 DNA. No significant association was found between integration of virus DNA and course of the disease. There was a trend towards an association between disease recurrence and the presence of the p53 codon 72 arginine homozygous genotype (OR = 3.41, p = 0.23). CONCLUSION: The physical state of HPV16 DNA does not seem to play a major role as a prognostic indicator in Hungarian cervical cancer patients, while the p53 codon 72 genotype may have an impact on the clinical outcome of the disease.

Poor clinical outcome in early stage cervical cancer with human papillomavirus-18 positive lymph nodes.

Epidemiologic and molecular studies have proven that human papillomavirus (HPV) plays an important role in the development of cervical cancer. However, the role of the virus in the progression of the disease, i.e. in the development of lymph node metastasis and in the adverse clinical outcome is poorly understood. We have been using the polymerase chain reaction (PCR) to study the presence and typing of human papillomavirus DNA since 1980 in cervical cancers and pelvic lymph nodes from the same patients. Out of the series of 47 cervical cancer patients we focused on four women (age: 41, 33, 35 and 56 years) in this article. The follow-up of these patients revealed early recurrences of the disease (7, 7, 17, 22 months) with very short survival (9, 10, 21, 24 months). Although we detected HPV-18 positivity both in the cervical tumors and in the regional lymph nodes too in all four cases, lymph nodes were negative by routine hystology in case of the three young patients (21, 33, 35 years of age). Our observations suggest that HPV type 18 positive cervical cancer patients, despite negative histological findings in the lymph nodes should be consider as a subpopulation for poor outcome especially in the young age group (p=0,022, Fisher's exact test).

Physical and chemical formations of lead contaminants in clay and sediment.

A possible sink for divalent lead in the environment is clays such as montmorillonite that have cation exchange capacities. To assess the reaction, a calcium-montmorillonite was mixed with lead perchlorate solutions of varying concentrations and at various pH's. The recovered solids were studied by a variety of techniques (X-ray photoelectron and infrared spectroscopy, scanning electron microscopy, and atomic force microscopy) to determine what, if any, alterations occurred. The ion exchange of lead for calcium reduced the hydrated water in the clay, and evidence for proton-lead ion exchange at the edges of the sheets was observed. Evidence for a second, unexpected, reaction was also observed. Small spots (0.2 to 1 microm) of lead enrichment were observed on the surface of clay particles. They were also observed on clays recovered from the sediment of a Hungarian lake. The results show that lead ions are adsorbed onto montmorillonite by two processes: cation exchange and nano- and microparticle production. Cation exchange leads to the even distribution of the ions, while the production of spots causes the enrichment of lead ions. The production of these particles is not expected from the thermodynamic properties of the solution and cannot be observed in the absence of clay.

[Follow-up of high risk, human papillomavirus (HPV)-positive patients with cancer of the uterine cervix]. / Magas kockázatú humán papillomavírus (HPV)-pozitív, méhnyakrákos betegek kórlefolyásának követése.

The study population consisted of 30 cervical cancer patients stage I.a-II.b. (FIGO) stages operated on according to the Wertheim technique. A parallel histological evaluation and HPV status determination were carried out on biopsies from the primary tumors and on the regional lymph nodes. A general primer mediated polymerase chain reaction (PCR) was performed at first and the samples not amplified were examined by type-specific primers. All except one primary tumors contained DNA-sequences characteristic for high risk HPV-types. The lymph nodes of these HPV-positive patients proved to be also HPV-positive with a frequency of 25/30 (83%). The frequency of the HPV-positivity was higher (100%) in the group of patients with HPV-18 positive status, than in the HPV-16 positive group. Two thirds of the evaluated regional lymph nodes were HPV-positive in the HPV-16 group of patients. The same HPV-types were harboured by the primary tumors and by the regional lymph nodes both in the HPV-16 positive and HPV-18 positive groups of patients. In the HPV-16-positive group of patients metastatic lymph nodes occurred with a frequency of 3/16, while the frequency of HPV-16 positivity in the same nodes was 11/15. In the group of patients with HPV-18 positivity the difference was even greater, 1/12 v. 12/12. Early recurrences were detected in a relation of 3 to 1 in the group of patients with histologically tumor-free and metastatic-positive lymph node status. At the same time all of the lymph nodes in this group with early recurrency (4/4) contained DNA-sequences characteristic for the HPV-18 type. These findings raise the hypothesis that the HPV-specific nucleic acids detected in the lymph nodes can be taken as sensitive indicators of metastases. The follow-up results support these hypothesis as patients with HPV-18 positive lymph node status showed early recurrencies and short survival that is poor prognosis not corresponding to the early stage of cervical cancer with histologically negative lymph node status.

[Prevalence of human papillomavirus infections in our five-year data]. / Humán papillomavírus-infekció prevalenciája közel ötéves anyagunkban.

Human papillomavirus infection proved to be the most important risk factor for the development of cervical cancer and its preblastomatosis. Human papillomavirus was detected from 1996. June to 2000. September at 1635 patients, who had been positive by colposcopy and/or cytology in an earlier examination. The place of the study were our outpatients' departments and consultations by specialists of Debrecen University, Department of Obstetrics and Gynecology. Hybrid capture system was used to demonstrate the presence of the virus and managed to prove it in the 28.9% of cases. 3.1% of the patients (51 persons) had acquired low-risk, and 23.6% (386 persons) high risk virus types, however 2.1% of the woman (35 patients) were infected with both low-risk and high-risk human papillomavirus types at the same time. Long time decrease of virus prevalence was observed after the age of 35 year, and the significant degree and timing decrease of after the age of 30 year at patients infected with combination of low-risk and high-risk virus types, respectively. This observation is indicative of the correlation between colposcopic-, cytologic abnormalities and the persisting high-risk human papillomavirus infections.

Granuloma gluteale infantum.

Granuloma gluteale infantum is an uncommon disorder, the pathogenesis of which remains unclear. Occlusion is a common factor and irritant napkin dermatitis is a frequent association. A six month old female infant with mild napkin dermatitis treated only with a dusting powder is presented.

Molluscum contagiosum virus: antibody responses in persons with clinical lesions and seroepidemiology in a representative Australian population.

An ELISA for molluscum contagiosum virus (MCV) was used to determine the antibody status of 35 adults with clinical infections and known human immunodeficiency virus (HIV) serology and of 357 persons (ages, 1 week-69 years) considered representative of the Australian population. MCV antibody was identified in 77% of persons with molluscum lesions: in 17 of 24 HIV-1-negative persons and in 10 of 11 who were HIV-1-positive. No relationship was evident between the serologic responses and the number of lesions or the duration of infection. The population survey revealed an overall seropositivity rate of 23%. The lowest antibody prevalence was in children aged 6 months to 2 years (3%), and seropositivity increased with age to reach 39% in persons >/=50 years old. These findings indicate that MCV infections, including very mild or subclinical cases, may be more common in the general community than previously suspected.