Optimized testing for C. trachomatis DNA in synovial fluid samples in clinical practice.

AIM: No standardized polymerase chain reaction (PCR) assay is available for detection of Chlamydia trachomatis (C. tr.) in synovial fluid (SF) for diagnostic use in clinical practice. This study tested the performance of two optimized molecular biology methods, to determine which is best suited for detecting C. tr. in SF clinical samples from patients with various rheumatologic diseases. METHODS: Two DNA extraction methods, i.e., (1) alkaline lysis and (2) QIAEX II Gel Extraction Kit® + cetyltrimethylammonium bromide (CTAB; Qiagen, Hilden, Germany), and C. tr.-omp1-152 bp PCR were tested in SF samples from a total of 329 patients with the following diagnoses: reactive arthritis (ReA; n = 10, 4 patients had posturethritic ReA), undifferentiated arthritis (UA; n = 66), rheumatoid arthritis (RA; n = 169), psoriatic arthritis (PSA; n = 12), and osteoarthritis (OA) n = 72. RESULTS: In SF samples, C. tr.-omp1-152 bp PCR in combination with alkaline lysis DNA extraction allowed detection of more C. tr.-positive samples: 3/10 (30%) ReA patients (all with posturethritic ReA) and 20/66 (38%) UA patients were positive, compared to the 0/10 (0%) patients with ReA and 1/66 (2%) with UA detected using the QIAEX II Gel Extraction Kit® + CTAB. Moreover, 2/12 (17%) SF samples from PSA patients tested positive with alkaline lysis. All samples from patients with OA and RA tested negative. CONCLUSION: Alkaline lysis in combination with C. tr.-omp1-152 bp PCR emerged as the most sensitive method for identification of C. tr. in clinical SF samples.

[Public health pests. Arthropods and rodents as causative disease agents as well as reservoirs and vectors of pathogens]. / Gesundheitsschädlinge. Arthropoden und Nagetiere als Krankheitsverursacher sowie Überträger und Reservoire von Krankheitserregern.

Globally, infectious diseases pose the most important cause of death. Among known human pathogenic diseases, approximately 50 % are zoonoses. When considering emerging infectious diseases separately 73 % currently belong to the group of zoonoses. In Central Europe, hard ticks show by far the biggest potential as vectors of agents of human disease. Lyme borreliosis, showing an estimated annual incidence between 60,000 and 214,000 cases is by far the most frequent tick-borne disease in Germany. Continually, formerly unknown disease agents could be discovered in endemic vector species. Additionally, introduction of new arthropod vectors and/or agents of disease occur constantly. Recently, five mosquito species of the genus Aedes have been newly introduced to Europe where they are currently spreading in different regions. Uncommon autochthonous transmission of dengue and chikungunya fever viruses in Southern Europe could be directly linked to these vector species and of these Ae. albopictus and Ae. japonicus are currently reported to occur in Germany. The German Protection against Infection Act only covers the control of public health pests which are either active hematophagous vectors or mechanical transmitters of agents of diseases. Use of officially recommended biocidal products aiming to interrupt transmission cycles of vector-borne diseases, is confined to infested buildings only, including sewage systems in the case of Norway rat control. Outdoor vectors, such as hard ticks and mosquitoes, are currently not taken into consideration. Additionally, adjustments of national public health regulations, detailed arthropod vector and rodent reservoir mapping, including surveillance of vector-borne disease agents, are necessary in order to mitigate future disease risks.

[Monitoring populations of rodent reservoirs of zoonotic diseases. Projects, aims and results]. / Monitoring von gesundheitsgefährdenden Nagetieren. Projekte, Ziele und Ergebnisse.

Rodents can harbor and transmit pathogens that can cause severe disease in humans, companion animals and livestock. Such zoonotic pathogens comprise more than two thirds of the currently known human pathogens. The epidemiology of some zoonotic pathogens, such as hantaviruses, can be linked to the population dynamics of the rodent host. In this case, during an outbreak of the rodent host population many human infections may occur. In other rodent-borne zoonotic diseases such phenomena are not known and in many cases the rodent host specificity of a given pathogen is unclear. The monitoring of relevant rodent populations and of the rodent-borne zoonotic pathogens is essential to (1) understand the distribution and epidemiology of pathogens and (2) develop forecasting tools to predict outbreaks of zoonoses. Presently, there are no systematic long-term monitoring programs in place for zoonoses in Germany. Rodent monitoring activities are largely restricted to the plant protection sector, such as for the common vole (Microtus arvalis) and forest-damaging rodents. However, during the last 10-15 years a number of specific research projects have been initiated and run for a few years and Norway rat (Rattus norvegicus) monitoring has been implemented in Hamburg and Lower Saxony. Based on close cooperation of federal and state authorities and research institutions these efforts could be utilized to gain information about the distribution and importance of rodent-borne zoonoses. Nevertheless, for the integration of rodent population dynamics and zoonotic disease patterns and especially for developing predictive models, long-term monitoring is urgently required. To establish a systematic long-term monitoring program, existing networks and cooperation need to be used, additional collaborators (e.g., pest control operators) should be included and synergetic effects of different scientific fields should be utilized.

[Rodenticide resistance and consequences]. / Rodentizidresistenz und Konsequenzen.

Resistance to anticoagulant rodenticides, such as warfarin was first described in 1958. Polymorphisms in the vitamin K epoxide reductase complex subunit 1 (VKORC1) gene and respective substitutions of amino acids in the VKOR enzyme are the major cause for rodenticide resistance. Resistant Norway rats in Germany are characterized by the Tyr139Cys genotype, which is spread throughout the northwest of the country. Resistant house mice with the VKOR variants Tyr139Cys, Leu128Ser and Arg12Trp/Ala26Ser/Ala48Thr/Arg61Leu (spretus type) are distributed over a number of locations in Germany. Resistance can reduce management attempts with consequences for stored product protection, hygiene and animal health. Anticoagulants of the first generation (warfarin, chlorophacinone, coumatetralyl) as well as bromadiolone and difenacoum are not an option for the control of resistant Norway rats. The same applies for house mice whereby the tolerance to compounds can be different between local incidences. Due to the higher toxicity and tendency to persist, the most potent anticoagulant rodenticides brodifacoum, flocoumafen and difethialone should be applied but only where resistance is known. In other cases less toxic anticoagulants should be preferred for rodent management in order to mitigate environmental risks. Resistance effects of further VKOR polymorphisms and their combinations, the spread of resistant rats and conditions supporting and reducing resistance should be investigated in order to improve resistance management strategies.

Low-flux hemodialysis suppresses interferon-gamma production: the possible role of beta2-microglobulin.

UNLABELLED: Interferon-gamma (IFN-gamma) is required for the acquired immune response involving maturation and activation of antigen-presenting cells (APC); both IFN-gamma production and activation of APC are impaired in ESRD patients on low-flux hemodialysis (HD). High levels of uremic toxins including beta2-microglobulin (beta2M) are thought to play a role in immunosuppression. PATIENTS AND METHODS: To test this hypothesis, we conducted an A-B-A crossover study to examine the influence of high-flux HD (A) versus low-flux HD (B) in ESRD patients (n = 14) using biocompatible synthetic dialyzer membranes (Polyamix(R), Gambro, Hechingen, Germany) and ultrafiltered bicarbonate-buffered dialysis fluid. Each study period lasted 6 months and at the end of each period, Kt/V urea, plasma levels of albumin, C-reactive protein (CRP) and beta2M were determined. In addition, production of IFN-gamma induced by heat-killed Staphylococcus epidermidis (Staph epi) was performed in whole blood cultures. RESULTS: Kt/V urea (mean +/- SEM) was 1.45 +/- 0.06 in Period A, 1.36 +/- 0.07* in Period B, and 1.51 +/- 0.07 in Period A' (*p < 0.01). beta2M resting levels increased from 26.6 +/- 1.42 mg/l in Period A to 34.7 +/- 2.30* mg/l in Period B, and decreased to 21.6 +/- 1.07 mg/l at the end of Period A' (*p < 0.0001). Albumin and CRP levels did not differ significantly. Staph epi-induced IFN-gamma production was 1.2 +/- 0.39 ng/1,000 PBMC at the end of Period A, 0.34 +/- 0.10 ng/1,000 PBMC* in Period B, and 2.71 +/- 0.67 ng/1,000 PBMC at the end of Period A' (*p < 0.05). There was an inverse correlation between b2M levels and whole blood IFN-gamma production (R2 = 0.26). CONCLUSION: High levels of uremic toxins such as beta2M suppress IFN-gamma production in ESRD patients under low-flux HD. High-flux HD reduces beta2M levels by 30% and improves IFN-gamma production suggesting an improved cellular immune response in ESRD patients.

Contact sensitization in patients with chronic wounds: results of a prospective investigation.

BACKGROUND: It is well known that patients with chronic wounds frequently acquire clinically relevant contact sensitizations to skin care products. OBJECTIVES: The aim of our study was to find out the actual frequency of contact sensitivities in patients with chronic wounds in Germany with particular attention to components of products used in modern wound therapy. METHODS: We examined the results of a prospective clinical investigation on skin patch tests of patients with chronic wounds. RESULTS: Altogether, 45 patients with chronic wounds were tested. In 25 (55.5%) of the examined patients, contact sensitization to at least one substance was detected. The most frequent contact sensitizations were to PVP-iodine (20%), balsam of Peru (15.6%) patients, fragrance mix (11.1%), colophony (8.8%) and potassium dichromate (6.7%). We also found sensitization to the wound dressings Varihesive (11.1%), Iruxol N (6.7%) and Comfeel (2.2%). CONCLUSIONS: We would like to propagate that therapists who are involved in wound treatment should also pay attention on the ingredients of applied modern wound dressings.

Effect of liposome-entrapped methotrexate on Ehrlich ascites tumor cells and uptake in primary liver cell tumor.

A therapeutically useful concentration (0,5 mg/ml) of Methotrexate was prepared in negatively charged artificial liposomes (lecithine: cholesterol: dicetylphosphate=5:5:1 molar ratios). Entrapment yield after separation on Sepharose 6B is nearly 100%. In contrast to free Methotrexate the liposome entrapped folic acid antagonist is eliminated only slowly by the kidneys and after intravenous application it is unevenly distributed in the body of mice, the highest concentrations being found in liver and spleen. Daily injections (7.5 mg/kg/day) of entrapped Methotrexate for 5 days into the tail vein of Ehrlich ascites tumor bearing mice reduced both tumor cell count and the production of ascites fluid about fourfold as compared to mice receiving the same dose of Methotrexate in the free form. Six hours after intravenous application of liposome entrapped Methotrexate the tissue concentration in normal liver is ca. 20-fold higher than when the same amount is applied in the free state. On the other hand, only a little uptake of liposome entrapped Methotrexate was detected in the tissue of nitrosamine-induced primary liver tumor when compared to normal liver tissue of rats.

Gabexate mesilate and camostate: new inhibitors of phospholipase A2 and their influence on the alpha-amylase activity in serum of patients with acute pancreatitis.

The new synthetic polyvalent protease inhibitors gabexate mesilate (ethyl-p[6-guanidinohexanoyloxy]-benzoate methansulfonate) and camostate (N,N-dimethylcarbamoylmethyl-4-[4-guanidinobenzoyloxy]-phenylacetate methansulfonate) were tested for possible inhibition of phospholipase A2 activity. In a pilot study, we treated 17 patients suffering from acute pancreatitis with continuous intravenous administration of gabexate mesilate, 450 mg/d. The results were compared with a placebo group (same standard therapy) of 21 patients suffering from acute pancreatitis. In vitro experiments showed that, at concentrations between 10(-4) and 5 X 10(-4) mol/L (depending on the enzyme assay employed) for gabexate mesilate and between 10(-3) and 5 X 10(-4) mol/L for camostate, a 50% reduction in phospholipase A2 activity was effected. Comparing the two groups of acute pancreatitis patients after 6 days of treatment with gabexate mesilate, we observed a statistically significantly lower alpha-amylase activity in the serum of treated patients compared with the placebo group.

Studies on the multiple forms of gamma-glutamyltransferase.

In sera of 53 patients with highly raised gamma-glutamyltransferase (gamma-GT) (EC 2.3.2.2) activity, the gamma-GT was separated by column chromatography on Sephadex DEAE A-50 into two fractions. In 35 sera only one gamma-GT fraction was found at a conductivity of 2.0--2.5 mS and in 18 sera two gamma-GT fractions were found at a conductivity of 0.2 mS and again at 2.0--2.5 mS. There was no opvious correlation to the underlying disease. In 20 sera the additional parameters, protein, protein electrophoresis, free and esterified cholesterol, bilirubin, triglycerides, lipoprotein pattern and lipoprotein X were examined. We recognized, that the two forms of the gamma-GT are not true isoenzymes but that the separation of the gamma-GT into two fractions is caused by a different distribution of the cholesterol containing lipoprotei,s.

[Diagnostic criteria for pancreatic sonography during the course of uncomplicated acute pancreatitis (author's transl)]. / Diagnostische Kriterien der Pankreassonographie im Verlauf einer unkomplizierten akuten Pankreatitis.

The course of uncomplicated pancreatitis was followed by sonography in 45 patients. The changes during the disease and their temporal relationship to the clinical stage are illustrated by an example. During the initial phase of the disease the sonographic findings may be so slight that they are easily missed, although clinical symptoms are present. It is only during the main phase of the disease that the typical sonographic findings of acute pancreatitis develop. At this time, maximal increase in amylase activity has usually passed. During recovery, with uninterrupted clinical and biochemical improvement, changes in the sonogram can still be seen for between three weeks and four months.

[Ultrasound in the diagnosis of ileitis terminalis Crohn (author's transl)]. / Sonographie in der Diagnostik der Ileitis terminalis crohn.

In order to check the value of sonography in the diagnosis of Crohn's disease, 81 patients who suffered from Crohn's disease were examined by means of real-time B-mode ultrasound. Bowel wall infiltration along the terminal ileum that could be shown to involve the coecum as well, appeared to be a reliable parameter in the diagnosis of Crohn's disease. This finding was most pronounced in acute disease whereas in chronic or mild disease, characteristic signs of Crohn's disease were absent. Among the complications of Crohn's disease formation of intraabdominal abscesses can be demonstrated by sonography. From our results we propose to employ diagnostic ultrasound in the acute stage of disease. After remission the diagnosis should be confirmed by endoscopy and X-ray examinations.

[Ulcer detection using 99m Tc-sucralfate]. / Ulkussuche mit 99mTc-Sucralfat.

Sucralfate is an agent commonly used in the therapy of gastrointestinal ulcers. It binds itself to proteins in the ulcer crater. A selective labeling with pure 99mTc-pertechnetate is possible. For an unequivocal ulcer localization or exclusion, images have to be taken in the upright and supine positions. Encouragement of gastric emptying is useful. Images should be taken at least twice, the last one 4 h after ingestion. The method might be used when endoscopy is difficult, incomplete or contraindicated, and for follow-up during therapy. We recommend it also in the search for the cause of intermittent bleeding.

Influence of a portacaval shunt on the distribution of 14C-chol-PC-DCP-liposomes and liposome entrapped 3H-methotrexate in the organs of rats.

The individual roles of hepatic parenchymal cells and non-parenchymal cells in the uptake of intravenously injected liposomes by the liver have not yet been clearly determined. Experimentally, it is suggested that a portacaval shunt reduces the phagocytic capacity of the liver RES. In our experiments a portacaval shunt does not influence the hepatic uptake of 14C-Chol-PC-DCP-liposomes and the liposome entrapped 3H-Methotrexate in rats. In normal rats and in rats with a portacaval shunt, most of the 14C- and 3H-radioactivity is found in the liver and the ratio between the amounts of free and liposome entrapped radioactivity 3 to 15 hours after i.v. injection is approximately 1:10. Therefore, it is concluded that the liver RES does not play an integral role in the enrichment of liposomes in the liver.

Epidemiology and treatment of gastric Campylobacter pylori infection: more questions than answers.

Two-hundred and ten consecutive patients undergoing routine gastroscopy were additionally investigated for evidence of Campylobacter pylori (C.p.). 106 patients were positive in one or more tests: 99.1% using a rapid urease detecting test (CLO-test), 80.2% histology, 78.3% cytology and 60% culture. We found no difference between the CLO-test results from biopsies taken from different parts of the stomach in individual patients. C.p. was found in 100% of patients with significant chronic antral gastritis, 67.7% with gastric ulcers, 65% with duodenal ulcers and in 12.1% of normal individuals. The C.p. infection was apparently eliminated in 50% of cases treated with bismuth subsalicylate (BSS) for four weeks. The combination of BSS with amoxicillin, tinidazole or an H2-receptor antagonist offered no advantage over BSS alone. Treatment with BSS led to improvement in symptoms and histological findings including healing of ulcers in patients with or without persistent C.p. infection. The recurrence of C.p. infection after apparently successful treatment was, however, 75% in 4 weeks. In conclusion, C.p. infection correlates strongly with the presence of chronic gastritis, and significantly with gastric and duodenal ulceration. The best diagnostic approach is the combination of a rapid urease detecting test and histology. C.p. infection is of long duration and difficult to eliminate. The most effective treatment for C.p. infection remains BSS as single agent.

Radio-opaque liposomes for the improved visualisation of focal liver disease by computerized tomography.

Liposomes laden with radio-opaque agents (ROA) were prepared by the reverse phase evaporation method. The liposome solutions--containing 1.7-2.5 mg iodine per mg lipid--were injected into Wistar rats. 60 min post injection the rats were scanned in a Siemens CT scanner. Following the injection of 260 mg encapsulated iodine per kg bodyweight--less than half the iodine of the normal ROA bolus injection--there was a long lasting arise in liver enhancement of 40 Hounsfield units (HU). In animals with liver tumours induced by nitrosamine lesions of 2-3 mm in diameter could be detected.

[Peppermint oil-caraway oil fixed combination in non-ulcer dyspepsia--comparison of the effects of enteric preparations]. / Pfefferminzöl/Kümmelöl-Fixkombination bei nicht-säurebedingter Dyspepsie-Vergleich der Wirksamkeit und Verträglichkeit zweier galenischer Zubereitungen.

223 patients with non-ulcer dyspepsia (dysmotility type dyspepsia or essential/idiopathic dyspepsia, also in combination with irritable bowel syndrome) were included in a prospective, randomised, reference- and double-blind controlled multicentre trial to compare two different preparations of a fixed combination of peppermint oil and caraway oil. The aim of the trial was to evaluate the equivalence of the efficacy and tolerability of these two preparations. The test formulation consisted of the drug combination in an enteric coated capsule containing 90 mg peppermint oil and 50 mg caraway oil, while an enteric soluble formulation containing 36 mg peppermint oil and 20 mg caraway oil was used as the reference. The main target item defined was the "difference in pain intensity between the beginning and the end of therapy", measured by the patient on a visual analogue scale (0 = no pain, 10 = extremely strong pain). In 213 patients (n = 108 on the test preparation, n = 105 on the reference preparation) with mean pain intensity baseline measurements of 6.1 points in the test preparation group and 5.9 points in the reference group a statistically significant decline in pain intensity was observed in the two groups (-3.6 resP. -3.3 points; p < 0.001; two-sided one-sample t-test). Equivalent efficacy of both preparations was demonstrated (p < 0.001; one-sided t-test for equivalence). With respect to concomitant variables, the results in both groups were also similar. Regarding "pain frequency", the efficacy of the test preparation was significantly better (p = 0.04; two-sided t-test for difference). Both preparations were well tolerated. Despite the higher dose, the adverse event "eructation with peppermint taste" was less frequent in the group treated with the test formulation, due to the enteric coated capsule preparation.

[Drug-induced pulmonary interstitial disease]. / Arzneimittelbedingte interstitielle Lungenerkrankungen.

HISTORY AND CLINICAL FINDINGS: Patient 1, a 72-year-old man, was admitted with a history of flu-like symptoms and dry cough for three weeks and exertional dyspnea. Prophylactic antibiotic treatment with nitrofurantoin (100 mg/day) had been given for one year for renal calculi. Patient 2, a 64-year-old man, was admitted with increasing dyspnea at rest. He was known to have several cardiological disorders, including episodes of ventricular tachycardia, for which he had been on amiodarone (200 mg/d) for 8 years. Both patients were in reduced general state of health. INVESTIGATIONS: In both patients the pO(2) was 68 mm Hg, in Patient 1 while breathing ambient air, in Patient 2 while breathing oxygen (15 l/min). Chest X-ray revealed bronchiectatic infiltrative changes bilaterally in the middle and lower lung fields in Patient 1. High-resolution computed tomography showed massive generalized interstitial infiltrates in the lung of Patient 2. TREATMENT AND COURSE: Nitrofurantoin and amiodarone, as potential causes of the pulmonary changes, were discontinued while steroid treatment was begun. This quickly resulted in improvement of symptoms and clinical findings. CONCLUSION: Drug induced pathogenesis has to be taken into account in the differential diagnosis of interstitial lung diseases. A causal association with the administered drugs is often difficult to establish. If suspicion is strong, treatment should be discontinued (replacement by a drug from another class). Depending on severity of clinical symptoms corticosteroids should also be given.