Comparison of Serum 25-Hydroxyvitamin D Levels in Patients With Malignant and Benign Gynaecological Disease.

BACKGROUND/AIM: Many studies have shown an antiproliferative, anti-inflammatory, anti-angiogenetic, and apoptosis-inducing effect of Vitamin D. A vitamin D deficiency has been associated with an increased risk for different types of cancer. This study examined vitamin D 25(OH)D levels in gynaecological cancers in comparison with benign gynaecological diseases. PATIENTS AND METHODS: Serum 25(OH)D levels in 688 gynaecological patients (488 with malignant, 200 with a benign gynaecological disease) were assayed between 2009 and 2015 using an electrochemiluminescence immunoassay. RESULTS: In total, the 25(OH)D levels in cancer patients were lower, but not significantly lower than those in cancer-free patients. Significant results were shown regarding seasonal effects for patients with breast-, endometrial and ovarian cancer. No significant effects occurred with regard to menopause status, nicotine, or grade in relation to 25(OH)D levels. CONCLUSION: 25(OH)D levels seem to influence gynaecological cancers.

Long-lasting cough in an adult German population: incidence, symptoms, and related pathogens.

Studies of the incidence of pertussis in adults have shown that it accounts for only 5-15% cases of prolonged coughing. We assessed the burden of suffering related to prolonged coughing and tried to identify further causative agents. Based on a sentinel study with 35 general practitioners in two German cities (Krefeld, Rostock), with 3,946 patients fulfilling the inclusion criteria, we estimated the incidence of prolonged coughing in adults. In 975 of these outpatients, PCR and/or serology for adenovirus, Bordetella pertussis and B. parapertussis, human metapneumovirus, influenza virus A and rhinovirus, parainfluenza virus, Mycoplasma pneumonia, and respiratory syncytial virus (RSV) were performed. Treatment data were extracted for a subgroup of 138 patients. Descriptive statistics, including Kaplan-Maier curves were generated. Yearly incidence ranged between 1.4 and 2.1% per population in the two cities. Adult patients sought medical attention only after a median of 3 weeks of coughing. Irrespective of smoking and unrelated to the identified pathogens, the median duration of coughing was 6 weeks, with an interquartile range of 4-11 weeks. In 48.3% of patients, possible pathogens were identified, among which adenovirus (15.1%), RSV (7.5%), B. pertussis (5.6%), and influenza viruses (4.0%) were most often found. Symptoms were not indicative of a specific agent and a total of 64% of patients received antibiotics. Prolonged adult coughing requiring medical attention prompts substantial healthcare use. Apart from B. pertussis, a broad range of pathogens was associated with the symptoms. However, patients sought medical attention too late to guide efficacious therapeutic interventions using the diagnostic tests.

The protective role of maternally derived antibodies against Bordetella pertussis in young infants.

In 20 infants with polymerase chain reaction-confirmed pertussis <6 months of age, median cord blood anti-pertussis toxin, anti-filamentous hemagglutinin and anti-pertactin IgG antibody values were lower than in 80 matched controls: 10.5 versus 13.5 anti-pertussis toxin IU/mL, 14.5/18.0 (anti-filamentous hemagglutinin) and 6.0/9.0 (anti-pertactin), respectively. Although differences of median (and mean) antibody values between groups were not significant, they are in line with the concept of infant protection by maternal antibodies and thus support the strategy of pertussis booster immunization in pregnant women.

Pulsed-field gel electrophoresis analysis of Bordetella pertussis isolates circulating in Europe from 1998 to 2009.

Between 1998 and 2009, Bordetella pertussis clinical isolates were collected during three periods, i.e., 1998 to 2001 (n = 102), 2004 to 2005 (n = 154), and 2007 to 2009 (n = 140), from nine countries with distinct vaccination programs, i.e., Denmark, Finland, France, Germany, The Netherlands, Norway, Poland, Sweden, and the United Kingdom. Pulsed-field gel electrophoresis (PFGE) analysis was performed according to standardized recommendations for epidemiological typing of B. pertussis. There were 81 different PFGE profiles, five of which (BpSR3, BpSR5, BpSR10, BpSR11, and BpSR12) were observed in 61% of the 396 isolates and shown to be predominant in almost all countries. The major profile, BpSR11, showed a decreasing trend from 25% to 30% in 1998 to 2005 to 13% in 2007 to 2009, and there were increases in BpSR3 and BpSR10 from 0% and 8% to 21% and 22%, respectively. One difference between these profiles is that BpSR11 contains isolates harboring the fim3-2 allele and BpSR3 and BpSR10 contain isolates harboring the fim3-1 allele. The total proportion of the five predominant profiles increased from 44% in 1998 to 2001 to 63% in 2004 to 2005 to 70% in 2007 to 2009. In conclusion, common PFGE profiles were identified in B. pertussis populations circulating in European countries with different vaccination programs and different vaccine coverages. These prevalent isolates contain the novel pertussis toxin promoter ptxP3 allele. However, there is evidence for diversifying selection between ptxP3 strains characterized by distinct PFGE profiles. This work shows that, even within a relatively short time span of 10 years, successful isolates which spread through Europe and cause large shifts in B. pertussis populations may emerge.

Maternally derived antibodies against Bordetella pertussis antigens pertussis toxin and filamentous hemagglutinin in preterm and full term newborns.

We measured IgG-anti-pertussis toxin and -IgG-anti-filamentous hemagglutinin antibody values in 43 full term and 34 preterm infants and 79 mothers. Antibody values were generally low and mean values were higher in full term than preterm infants. Transfer ratios increased with gestational age of the newborns. Based on these findings, maternal and neonatal immunization strategies should be considered to protect young infants from pertussis.

Characterization of reference materials for human antiserum to pertussis antigens by an international collaborative study.

Enzyme-linked immunosorbent assay (ELISA) has been widely used to evaluate antibody responses to pertussis vaccination and infection. A common reference serum is essential for the standardization of these assays. However, no internationally recognized reference serum is available. At the request of the Expert Committee on Biological Standardization (ECBS) of the World Health Organization (WHO), a set of four candidate international standards has been prepared. These candidate materials have been assessed for suitability and compared to the widely used U.S. reference pertussis antiserum (human) lot 3, lot 4, and lot 5 by 22 laboratories from 15 countries in an international collaborative study. Laboratories measured immunoglobulin G (IgG) and IgA antibodies to pertussis toxin (PT), filamentous hemagglutinin (FHA), pertactin (PRN), and fimbriae (Fim2&3) using their established immunoassays. The results of this study showed each of the four candidates to be suitable as an international standard. With the agreement of the participants, a recommendation has been made to the ECBS that the candidate material coded 06/140 be established as the First International Standard for pertussis antiserum (human), with the following assigned international units (IU): IgG anti-PT, 335 IU/ampoule; IgA anti-PT, 65 IU/ampoule; IgG anti-FHA, 130 IU/ampoule; IgA anti-FHA, 65 IU/ampoule; IgG anti-PRN, 65 IU/ampoule; and IgA anti-PRN, 42 IU/ampoule. No formal units have been proposed for anti-Fim2&3 because most assays used a mixture of fimbrial antigens. In addition, the candidate material coded 06/142 has been proposed as a WHO working preparation for characterization of assay systems.

Differences of humoral and cellular immune response to an acellular pertussis booster in adolescents with a whole cell or acellular primary vaccination.

To study the pertussis-specific immune response of adolescents with different prevaccination schedules, we measured the humoral and cell-mediated immunity (CMI) to pertussis antigens before and after a five-component Tdap booster vaccination in 78 adolescents, who had previously received either five doses of a two-component acellular pertussis vaccine (aP; last dose age 4-6 years), four doses of aP (last dose age 18-24 months), or four doses of whole cell pertussis vaccine (wcP; last dose age 18-24 months). The proportion of participants with a twofold rise in titre was 79% against pertussis toxin (PT), 94% against filamentous hemagglutinin (FHA), and 99% against pertactin (PRN) without significant differences between the three groups. However, participants with primary wcP vaccination showed higher postvaccination titres to pertussis toxin (geometric mean titre, GMT 50.3EU/ml) than those with either four (GMT 17.1EU/ml) or five (GMT 16.4EU/ml) previous aP doses. CMI indices to PT, FHA, PRN and fimbriae (FIM) increased after vaccination and were similar between groups. The current adolescent Tdap booster immunization induced good humoral and cellular immune response to pertussis. The higher antibody titres to pertussis toxin may indicate a more effective priming of B cell memory after primary whole-cell vaccination.

Cost-effectiveness of adult pertussis vaccination in Germany.

BACKGROUND: The incidence of pertussis in adults is high despite good childhood vaccination coverage. An adult formulation of an acellular pertussis vaccine is licensed and available for use in Germany. OBJECTIVE: To evaluate the potential health benefits, risks, costs and cost-effectiveness of routine pertussis vaccination programs for German adults. METHODS: A Markov model was used to simulate health states and immunity levels associated with pertussis disease and vaccination. The following strategies were evaluated: (1) no adult pertussis vaccination, (2) one-time adult vaccination at 20-64 years, and (3) adult vaccination with decennial boosters. Our main outcome measures were costs (2006 Euros), cases prevented, incremental cost per case prevented and incremental cost per quality-adjusted life year (QALY) saved. We performed sensitivity analyses for key assumptions in the model including disease incidence, vaccine cost, vaccine efficacy, disease costs and frequency of adverse events. Future costs and benefits were discounted at 3%. RESULTS: At a disease incidence of 165 per 100,000, the one-time adult vaccination strategy would prevent 498,000 cases, and the decennial adult vaccination strategy would prevent 1 million cases. Approximately 31 million adults ( approximately 62% of the cohort) would be vaccinated with a one-time adult vaccination strategy for a total program cost of 366 million Euros, while a decennial vaccination strategy would cost 687 million Euros. The one-time adult vaccination strategy resulted in CE ratios of 5800 Euros per QALY saved, or 160 Euros per pertussis case prevented. The decennial booster strategy cost 7200 Euros per QALY saved, or 200 Euros per case prevented. The results were most sensitive to assumptions about disease incidence and vaccine cost. CONCLUSIONS: Routine vaccination of German adults aged 20-64 years with Tdap is cost-effective.

Long-term immunogenicity of a single dose of acellular pertussis vaccine in paediatric health-care workers.

OBJECTIVE: Monitor the long-term immunogenicity of a single dose of acellular pertussis vaccine in health-care workers. DESIGN: German health-care workers and child-care workers received a single dose of a monovalent acellular pertussis vaccine (PAC-Mérieux) in an open-label study. Blood samples were taken before (n=261), 4 weeks after (n=246), 1 year (n=187), 2 years (n=53), 3 years (n=134) and 4 years (n=37) after vaccination. IgG- and IgA-anti-pertussis-toxin (PT), IgG- and IgA-anti-filamentous hemagglutinin (FHA), and IgG-anti-pertactin (PRN) were measured by ELISA. RESULTS: Of all subjects, 97.1%, 99.2% and 97.2% had an antibody response to PT, FHA and PRN, respectively. Four weeks after vaccination the median titres of IgG antibodies to PT, FHA and PRN were 314, 785 and 84 EU/l, respectively, and all vaccinees had an immune response to at least one pertussis antigen. IgA-anti-PT and IgA-anti-FHA responses were found in 63.4% and 96.3% of subjects with a median titre of 30 and 196 EU/ml, respectively. The titre of IgG-anti-PT decreased slowly with a median concentration of 76, 71, 71 and 63 EU/ml after 1, 2, 3 and 4 years, respectively. Secondary titre increases were observed in 0.5%, 3.3%, 5.6% and 12.5% of the vaccinees 1, 2, 3, and 4 years after vaccination. CONCLUSION: In German health paediatric care workers long-lasting immune responses with high antibody levels could be induced by a single dose of acellular pertussis vaccine. A renewed contact with B. pertussis antigens resulted in a measurable immune response to PT between 0.5% (1 year p.v.) and 12.5% (4 years p.v.).

Measuring immunoglobulin g antibodies to tetanus toxin, diphtheria toxin, and pertussis toxin with single-antigen enzyme-linked immunosorbent assays and a bead-based multiplex assay.

Bead-based assay systems offer the possibility of measuring several specific antibodies in one sample simultaneously. This study evaluated a vaccine panel of a multianalyte system that measures antibodies to tetanus toxin, diphtheria toxin, and pertussis toxin (PT) from Bordetella pertussis. The antibody concentrations of human immunoglobulin G (IgG) to PT, tetanus toxin, and diphtheria toxin were measured in 123 serum pairs (total of 246 sera) from a vaccine study. The multianalyte bead assay was compared to a standardized in-house IgG- anti-PT enzyme-linked immunosorbent assay (ELISA) of the German reference laboratory for bordetellae, as well as to various commercially available ELISAs for anti-PT IgG, anti-tetanus IgG, and anti-diphtheria IgG. The results of the multiplex assay regarding the antibodies against diphtheria toxin compared favorably with a regression coefficient of 0.938 for values obtained with an ELISA from the same manufacturer used as a reference. Similarly, antibodies to tetanus toxin showed a correlation of 0.910 between the reference ELISA and the multianalyte assay. A correlation coefficient of 0.905 was found when an "in-house" IgG anti-PT and the multiplex assay were compared. Compared to single ELISA systems from two other manufacturers, the multiplex assay performed similarly well or better. The multianalyte assay system was a robust system with fast and accurate results, analyzing three parameters simultaneously in one sample. The system was well suited to quantitatively determine relevant vaccine induced antibodies compared to in-house and commercially available single-antigen ELISA systems.

Pertussis: not only a disease of childhood.

INTRODUCTION: Pertussis is not just a childhood disease, but a respiratory infection that causes persistent cough in all age groups, from newborns to the elderly. METHODS: The authors performed a selective literature search and reviewed national and international recommendations for treatment and vaccination. RESULTS: Pertussis is found principally in young unvaccinated infants, but school-age children, adolescents, and adults are also affected. Up to 1% of infants contract pertussis, and their respiratory symptoms are often accompanied by apnea. School-age children occasionally display the coughing spasms typical of the disease. Annually, 0.2% to 0.5% of all adolescents and adults are infected and suffer from prolonged, frequently non-paroxysmal coughing. Severe and fatal cases of pertussis occur mainly in newborns and infants, and 25% of affected adults experience complications. Bordetella DNA may be detected by polymerase chain reaction (PCR) for four weeks after symptom onset; except in infants, the sensitivity of this diagnostic technique is low. Although the diagnosis can be confirmed by serological tests, the methods are not well standardized. Treatment with a macrolide prevents the spread of infection, but generally does not alleviate the symptoms. Combination vaccines are the most effective means of prophylaxis. DISCUSSION: Pertussis is usually not included in the differential diagnosis of persistent respiratory symptoms. The considerable burden of disease could be reduced in adults and young infants by vaccinating adults with acellular combination vaccines.

Bordetella pertussis strains circulating in Europe in 1999 to 2004 as determined by pulsed-field gel electrophoresis.

Clinical isolates of Bordetella pertussis collected during the year 2004 (n = 153) in eight European countries, Denmark, Finland, France, Germany, The Netherlands, Poland, Sweden, and United Kingdom, were analyzed by pulsed-field gel electrophoresis (PFGE), and their PFGE profiles were compared with those of isolates collected in 1999 (n = 102). The 255 isolates produced 59 distinct PFGE profiles. Among the 153 isolates from 2004, 36 profiles were found, while within the 102 isolates from 1999, 33 profiles were detected. One PFGE profile, BpSR11, was dominant (30% to 50%) in all countries except Denmark (10%) and Poland (0%). In comparison with 1999, there was an increase in BpSR11 prevalence in Finland in 2004 from 5% to 40%, coinciding with a major incidence peak. Some other PFGE profiles seemed to be associated with limited dissemination. Poland was the only country in which the most common actual European PFGE profiles were not found. In a dendrogram analysis, all common PFGE profiles were identified within PFGE group IV, and BpSR11 clustered together with PFGE subgroup IVbeta. Compared to the 1999 isolates, PFGE group V representative for pertactin variant prn3 strains had disappeared, and a new cluster was seen. In conclusion, some PFGE profiles, such as BpSR11, evidently have a higher capacity to spread, suggesting increased fitness to the present immunological environment. It is therefore of major interest to continue with surveillance programs of B. pertussis isolates, as both waning vaccine-derived immunity and strain variation may play a role in the persistence of pertussis.

Anticancer drugs induce mdr1 gene expression in recurrent ovarian cancer.

Ovarian cancer is currently the most lethal gynecologic malignancy in Europe and the US. Platin analogues and paclitaxel demonstrate high remission rates, but unfortunately the efficacy of cytostatic agents is limited by the development of multidrug resistance (mdr). Clinical paclitaxel resistance is often associated with mdr1 overexpression. In a recent study, we introduced a highly specific quantitative real-time reverse transcriptase polymerase chain reaction for the quantification of mdr1 transcripts. In the present study, we demonstrate that primary tumor cells from patients with recurrent ovarian cancer overexpress mdr1. To evaluate mdr1 expression, we collected tumor cells from 77 ovarian cancer patients (13 chemotherapy-naive ovarian cancer, 64 recurrent ovarian cancer). Cancer cells were aspirated from 49 solid specimens (63%) and 28 ascitic fluids (37%). Subsequently, cancer cells were exposed in 221 short-term cultures either to blank medium (control) or to a single anticancer drug, cisplatin, doxorubicin or paclitaxel. The drug concentrations applied referred to clinical relevant doses. mdr1 mRNA expression was significantly higher in specimens from recurrent ovarian cancer incubated in paclitaxel than in specimens from chemotherapy-naive ovarian cancer. No significant differences were detectable between the mean value of mdr1 mRNA expression in tumor specimens from recurrent ovarian cancer incubated in cisplatin or doxorubicin. Differences within the untreated patients group were also not statistically significant. The result of this study confirms clinical observations, as well as in-vitro studies based on tumor cell lines, that paclitaxel resistance is correlated with mdr1 overexpression.

Immunogenicity of a single dose of reduced-antigen acellular pertussis vaccine in a non-vaccinated adolescent population.

German adolescents (n=123) without previous pertussis vaccination, no history of pertussis and low IgG-anti-pertussis-toxin (PT) levels received one dose of the Tdap vaccine Boostrix. Blood samples were taken before, and 5-12 days and 29-49 days after vaccination. IgG- and IgA-anti-PT, IgG- and IgA-anti filamentous hemagglutinin, IgG-anti-pertactin, IgG-anti-tetanus-toxin, and IgG-anti-diphtheria-toxin were measured by ELISA. 88.6% of subjects had an immune response to PT, and all vaccinees had an immune response to at least one pertussis antigen 29-49 days after vaccination. IgA-anti-PT and IgA-anti-FHA responses were found in 43 and 81% of subjects, respectively. This study shows that in unvaccinated German adolescents pertussis immunity can be achieved by a single dose of Tdap.

Pulsed-field gel electrophoresis analysis of Bordetella pertussis populations in various European countries with different vaccine policies.

The increasing incidence of pertussis in a number of countries, despite good vaccination coverage, is a cause for concern. We used pulsed-field gel electrophoresis (PFGE) typing to examine the genetic diversity of 101 clinical isolates of Bordetella pertussis, recovered during 1999-2001, and circulating in five different European countries to evaluate temporal and geographical distribution. This DNA fingerprinting approach seems to be a more discriminative epidemiological tool than sequencing of individual genes. Despite differences in vaccination policies in the five countries, these European isolates were found to be very similar and fell into the same major PFGE profile groups, with a predominance of one profile group. There was no evidence of geographic clustering, except that one new profile subgroup was predominantly found in one country. This study provides a baseline for continued surveillance of the B. pertussis population in Europe.

Fatal influenza A virus infection in a child vaccinated against influenza.

We report a fatal infection with influenza A virus in a 13-year-old child who had been vaccinated against influenza. Influenza A virus RNA was detected by PCR in lungs, bronchi and myocardium. A penicillin-sensitive strain of Staphylococcus aureus was also isolated from her bronchi. This case indicates that a primary viral pneumonia with influenza A virus complicated by a bacterial superinfection with S. aureus can run a fatal course even in a vaccinated child.

Cisplatin, doxorubicin and paclitaxel induce mdr1 gene transcription in ovarian cancer cell lines.

The clinical observation of the multidrug resistance (MDR) phenotype is often associated with overexpression of the mdrl gene, in particular with respect to ovarian cancer. However, until now the mdrl-inducing potential of commonly used antineoplastics has been only incompletely explored. We performed short-term cultures of six ovarian cancer cell lines (MZOV4, EF027, SKOV3, OAW42, OTN14, MZOV20) exposed to either blank medium or cisplatin, doxorubicin or paclitaxel at concentrations related to the clinically achievable plasma peak concentration. A highly specific quantitative real-time RT-PCR was used to detect the Mdr1 transcripts. Mdrl mRNA contents were calibrated in relation to coamplified GAPDH mRNA. Mdrl mRNA was detectable in each cell line. In 13 out of 18 assays (72%) the specific anticancer drug being tested induced mdr1 transcription. No decrease in mdr1 mRNA concentration was observed. Our data suggest that mdr1 induction by antineoplastics is one of the reasons for failure of ovarian cancer therapy but may vary individually.

Antibodies to pertussis antigens in pediatric health care workers.

BACKGROUND: To compare the antibody concentrations against Bordetella antigens in health care workers in a pediatric hospital with those of two different populations without professional contact with children. METHODS: In a pediatric hospital 155 health care workers (135 female, 20 male), 292 male navy recruits after 3 months at sea and 146 regular blood donors (41 female, 105 male) were screened for antibodies of isotypes IgG and IgA to pertussis toxin (PT) and filamentous hemagglutinin (FHA) by enzyme-linked immunosorbent assay. RESULTS: Pediatric health care workers were positive for IgG anti-PT in 88%, for IgA anti-PT in 52%, for IgG anti-FHA in 99% and for IgA anti-FRA in 84%. Relative numbers for blood donors and recruits were 86 and 80% for IgG anti-PT, 56 and 55% for IgA anti-PT, 100 and 98% for IgG anti-FHA and 92 and 82% for IgA anti-FHA, respectively. Reverse cumulative distribution of all antibodies except for IgA anti-FHA showed no differences among the three groups; 2% of pediatric personnel, 3% of blood donors and 3% of navy recruits, respectively, had IgG anti-PT > or = 100 enzyme-linked immunosorbent assay units/ml, indicating a recent contact to Bordetella pertussis. CONCLUSION: Antibodies to B. pertussis antigens, such as IgG/IgA anti-PT and IgG/IgA anti-FHA, were similarly distributed in all three groups. Our results suggest that exposures leading to measurable immune responses to pertussis antigens in German pediatric health care workers are not significantly more frequent than in other populations without professional contacts with children.

Real-time LightCycler PCR for detection and discrimination of Bordetella pertussis and Bordetella parapertussis.

Real-time PCR assays based on the LightCycler technology were developed for individual (simplex PCR) and simultaneous (duplex PCR) detection and discrimination of Bordetella pertussis and Bordetella parapertussis in clinical samples. The assays were evaluated with 113 specimens from patients with and without symptoms of pertussis. Results were compared to those from conventional culture and TaqMan real-time PCR. The analytical sensitivity ranged from 0.1 to 10 CFU for B. pertussis and B. parapertussis, and intra- and interassay variations were less than 7%. Results were available within 2 h. With the simplex format, 21 of 100 samples from patients with clinical symptoms of pertussis were positive for B. pertussis and/or B. parapertussis. With the duplex format, 18 of 100 samples were positive. LightCycler PCR increased the diagnostic sensitivity over that of culture by 2.0-fold (duplex PCR) (P = 0.08) to 2.3-fold (simplex PCR) (P = 0.02). Our data suggest that duplex PCR in this format showed good analytical sensitivity but lost some sensitivity on clinical samples compared with the simplex format.

Evaluation of a real-time PCR assay for detection of Bordetella pertussis and B. parapertussis in clinical samples.

A real-time PCR assay based on the TaqMan technology was developed for the detection of Bordetella pertussis and B. parapertussis in clinical samples. The assay was evaluated with 182 specimens from 153 patients with and without symptoms of pertussis. The analytical sensitivity ranged from 0.1 to 10 cfu for B. pertussis and B. parapertussis, respectively, and diagnostic sensitivity was 94.1% when culture was used as a reference. No sample from a patient without symptoms of pertussis was positive in PCR. Twenty-four of 28 patients who were negative by culture and positive by PCR assay met the CDC clinical case definition for pertussis; the remaining four patients had paroxysms of shorter duration. Intra- and inter-assay variation were <5% and results were available within 4 h.