A German Chlamydia trachomatis screening program employing semi-automated real-time PCR: results and perspectives.

BACKGROUND: Genital Chlamydia trachomatis infection is a worldwide public health burden. A screening program for C. trachomatis was therefore initiated by the public health insurers in Germany ("Gemeinsamer Bundesausschuss", GBA) in April 2008. OBJECTIVES: To estimate C. trachomatis prevalence from screening 115,766 asymptomatic females and 20,033 female patients with unspecific abdominal pain. STUDY DESIGN: Urine samples (pooled by five for the asymptomatic screening subjects) and cervical swabs were analyzed using semi-automated real-time PCR. Infection prevalence was determined separately in four categories of women, defined by health status (asymptomatic screening vs. non-screening with unspecified symptoms) and test material used. Comparative analyses were stratified by age and pregnancy status. RESULTS: Experimental evaluation of the assay used revealed a detection limit of 379 genome copies/ml urine. For pooled urine samples, the positive predictive value was 100% whereas the negative predictive value equaled 98.1%. The observed infection prevalence was higher for cervical swabs than for urine samples. Prevalence estimates also differed significantly between pregnant and non-pregnant adolescents (< or = 20 years), irrespective of the test material used (10.2% vs. 7.3% for cervical swabs, 10.9% vs. 6.1% for pooled urine samples). CONCLUSIONS: Our retrospective study, based upon a very large number of females from all parts of Germany, revealed a high infection prevalence in adolescents, particularly in pregnant adolescents, thereby justifying the screening directive of the German GBA.

Cell-dependent influence on the phagocytosis induced by non-ionic contrast medium injection.

Iodinated contrast media (CM) can inhibit phagocytosis. To better understand the importance of this effect upon the elementary defence mechanism, the aim of the study was to compare the in vivo effect of non-ionic CM on the engulfing ability of peripheral blood phagocytic cells from patients undergoing CM-enhanced CT. Neutrophil granulocytes and monocytes from patients' peripheral blood obtained before and 30 min after CM injection were incubated with fluorescent-labelled Escherichia coli bacteria. Both the percentage of cells that engulfed bacteria and the phagocytic activity per cell has been determined by flow cytometry. We found that phagocytosis was greater in neutrophils than in monocytes. CM decreased the percentage of monocytes phagocyting bacteria, both at 4 degrees C (20.3%+/-3.3% versus 16.1%+/-2.0%; p<0.03) and at 37 degrees C (51.6%+/-4.1% versus 47.5%+/-2.6%; p>0.05), and increased the percentage of neutrophils at 4 degrees C (11.8%+/-2.1% versus 14.3%+/-2.2%; p<0.002) and at 37 degrees C (83.1%+/-3.6% versus 85.1%+/-3.2%; p>0.05). The phagocytic activity decreased significantly at 37 degrees C in monocytes (p<0.02), and was not affected in neutrophils. CM injection has different effects on both the percentage of phagocytosing cells and the phagocytic activity in monocytes and neutrophils. The inhibitory effect on monocyte phagocytosis seems to be compensated by neutrophils.

[Combination of radiological and nuclear medical imaging in animals: an overview about today's possibilities]. / Kombinierte radiologische und nuklearmedizinische Bildgebung in Tierexperimenten: Ein Uberblick über die aktuellen Möglichkeiten.

Molecular imaging of small animals has made considerable progress in the last years. Various research fields are interested in imaging small animals due to the lower numbers of animals per experiment. This has advantages with respect to financial, ethical and research aspects. Non-invasive imaging allows examination of one animal several times during the same experiment. This makes it possible to follow a pathological process in the same animal over time. However, the radiological methods used such as magnetic resonance imaging or computed tomography as well as the nuclear medicine methods such as single photon emission computed tomography or positron emission tomography suffer from disadvantages. Molecular aspects are limited in the radiological methods while anatomical localization is difficult in nuclear medicine. The fusion of these methods leads to additional information. This review shows today's possibilities with their advantages as well as disadvantages.

[Molecular imaging of apoptosis in cardiovascular diseases]. / Molekulare Bildgebung der Apoptose bei kardiovaskulären Erkrankungen.

Molecular imaging of functional parameters such as apoptosis (programmed cell death) in vivo opens new possibilities in clinical diagnostic and scientific research. Especially in the case of cardiovascular diseases that are mainly responsible for both morbidity and mortality in Western industrial nations, innovative non-invasive examination strategies are necessary for early diagnosis of these diseases. Since apoptosis unlike necrosis is present even after minor alterations of the microenvironment of cells and has been shown to be involved in a large number of cardiovascular diseases, there are currently several experimental studies underway with the goal of imaging apoptosis in vivo. The review discusses the basics of apoptosis in myocardial infarction, myocarditis, atherosclerosis, restenosis after angioplasty and stent implantation, currently used imaging techniques, achieved results, and future possibilities for molecular imaging of apoptosis.

[The apoptosis marker enzyme poly-(ADP-ribose) polymerase (PARP) in systemic lupus erythematosus]. / Das Apoptose-Markerenzym Poly-(ADP-Ribose-)Polymerase (PARP) beim systemischen Lupus erythematodes.

The enzyme poly-(ADP-ribose) polymerase (PARP) is localized within the cell nucleus and catalyzes DNA-repair. During programmed cell death (apoptosis), PARP is enzymatically cleaved. Detection of the cleavage products is characteristic for apoptosis. In patients with systemic lupus erythematosus (SLE), the highly ordered signal transduction cascade of apoptosis is disturbed. SLE patients show reduced PARP activity . PARP cleavage products are mainly found in association with either antinuclear and/or anti-dsDNA antibodies. In addition, serum samples from SLE patients and other autoimmune diseases display anti-PAR and anti-PARP autoantibodies.

[Molecular imaging of apoptosis and necrosis -- basic principles of cell biology and use in oncology]. / Molekulare Bildgebung von Apoptose und Nekrose -- Zellbiologische Grundlagen und Einsatz in der Onkologie.

Soon molecular imaging techniques will play a prominent role in basic scientific research and clinical approaches. In particular, important aspects of medicine such as apoptosis and gene- and stem-cell therapy will play a pivotal role in radiology too. This review presents the basic principles of apoptosis, recent results and future perspectives of apoptosis imaging. Apoptosis or programmed cell death is a precisely regulated, complex cascade of molecular events to eliminate individual cells. Disturbances may lead to diseases like malignancies and neurodegenerative diseases that are of clinical relevance. Several therapeutic strategies in oncology are based on apoptosis induction; conversely, resistance to therapy is indicative of decreased apoptosis induction. Whereas up to now the clinician had to depend exclusively on biopsy specimens to detect apoptosis, the feasibility of non-invasive imaging of this cell-biological phenomenon in vivo opens up new horizons in future. This review focuses on different modifications of this imaging technique, with and without the use of molecular probes (e. g. annexin V, synaptotagmin I), in vitro and in vivo using the various detector systems (like MRI, flow cytometry) currently available. Future perspectives are also addressed.

[Non-ionic iodinated dimeric versus monomeric X-ray contrast media: effects on complement factors in vivo]. / Nichtionische iodierte dimere versus monomere Röntgenkontrastmittel: Effekte auf Komplementfaktoren in vivo.

PURPOSE: To survey contrast media (CM)-induced alterations of complement factors. MATERIAL AND METHODS: In 31 adult patients, who received either an iotrolan (n = 19) or iopromide (n = 12) i. v. injection for CT examination, complement factors C1 q, C3, C4, C5 a, and C1-esterase inhibitor in serum/plasma samples were analyzed. The samples were obtained prior to and 5 min., 30 min., 1 hr., 6 hrs. and 24 hrs. after CM injection. RESULTS: 5 patients (16.1 %) developed a CM reaction. 4 of these were patients who received iotrolan. Other than minimal data, we neither found a significant influence of the CM on complement activation nor a difference between the analyzed CM. In detail, 5 min. after CM administration, we found the tendency to be for the values to decrease and then to return to the basic value. The changes induced by iotrolan were more pronounced than those induced by iopromide; nevertheless the differences were not statistically significant. A more pronounced decrease of C3 and C4 after iotrolan injection indicates the activation of the classic way, while this could not been observed after iopromide injection. One patient who experienced an unwanted reaction towards iotrolan showed shifts of C1 q, C1 INH, C3 and C4. CONCLUSION: The presented data shows different influences of CM injection on the analyzed complement factors after 5 min. that were commonly no longer present 30 min. after CM injection. The dimeric iotrolan induced a significantly increased frequency of unwanted CM reactions than the monomeric iopromide. The question of whether iotrolan is possibly able to activate the classic way of the complement cascade should be analyzed in the future in a greater patient group.

Apoptosis: the link between autoantibodies and leuko-/lymphocytopenia in patients with lupus erythematosus.

OBJECTIVE: To analyse the relation between serum autoantibodies and leuko-/lymphocytopenia in patients with lupus erythematosus (LE). METHODS: Laboratory routine analyses (white blood cell counts, autoantibody detection), and flow cytometry (annexin V, CD3, CD4, CD8, CD95, F(ab)2 anti-human IgG) have been performed in LE-patients versus healthy controls. In vitro, the influence of pooled serum containing anti-dsDNA antibodies has been analysed on the CD95 expression. RESULTS: Leukocytes, lymphocytes, CD3+, CD3+ CD4+, and CD3+ CD8+ cells from LE-patients were significantly reduced compared with controls. Patients with autoantibodies had significantly lower absolute cell counts than those without. Apoptosis was increased in LE versus controls (p < 0.04). The percentage of CD95+ T-cells was increased, and the absolute number of CD95+ cells was reduced in LE. In vitro up-regulation of CD95 could be detected on T-cells of healthy donors. Induction of CD95 seems to be donor-dependent. CONCLUSION: The data suggest that autoantibodies may be associated with blood cytopenias. CD95 seems to play a central role in this signalling cascade. The underlying mechanism is unclear, but seems to be autoantibody-related apoptosis induction.

Engineering a polyketide with a longer chain by insertion of an extra module into the erythromycin-producing polyketide synthase.

BACKGROUND: Modular polyketide synthases catalyse the biosynthesis of medically useful natural products by stepwise chain assembly, with each module of enzyme activities catalysing a separate cycle of polyketide chain extension. Domain swapping between polyketide synthases leads to hybrid multienzymes that yield novel polyketides in a more or less predictable way. No experiments have so far been reported which attempt to enlarge a polyketide synthase by interpolating additional modules. RESULTS: We describe here the construction of tetraketide synthases in which an entire extension module from the rapamycin-producing polyketide synthase is covalently spliced between the first two extension modules of the erythromycin-producing polyketide synthase (DEBS). The extended polyketide synthases thus formed are found to catalyse the synthesis of specific tetraketide products containing an appropriate extra ketide unit. Co-expression in Saccharopolyspora erythraea of the extended DEBS multienzyme with multienzymes DEBS 2 and DEBS 3 leads to the formation, as expected, of novel octaketide macrolactones. In each case the predicted products are accompanied by significant amounts of unextended products, corresponding to those of the unaltered DEBS PKS. We refer to this newly observed phenomenon as 'skipping'. CONCLUSIONS: The strategy exemplified here shows far-reaching possibilities for combinatorial engineering of polyketide natural products, as well as revealing the ability of modular polyketide synthases to 'skip' extension modules. The results also provide additional insight into the three-dimensional arrangement of modules within these giant synthases.

Revision surgery after failed unicompartmental knee arthroplasty: a study of 35 cases.

This retrospective study evaluates the results of 35 revision procedures after failed unicompartmental knee arthroplasty (UKA) in 34 patients, which were done during the period 1986 to 1996. There were 28 women and 6 men with a mean age of 71 years (range, 54-85 years). In all cases, St. Georg and Endo (W. Link, Hamburg, Germany) unicompartmental prostheses were used except 1 PCA unicondylar implant (How medica, Rutherford, NJ) and 1 Böhler unicondylar implant (Allo Pro, Baar, Switzerland). Failures most frequently were due to aseptic loosening followed by polyethylene wear. Two deep infections occurred. Revisions were performed 1 week to 11 years after UKA; 23 were required within the first 5 years. In most cases, revision was to a total knee arthroplasty. Partial component exchange was done in 9 cases. All 34 patients were evaluated clinically after exchange arthroplasty. After a mean follow-up time of 4 years (range, 1-12.2 years), we found 11 excellent, 13 good, 4 fair, and 7 poor results according to the Hospital for Special Surgery score. The fair and poor results were due to aseptic loosening of the knee prosthesis in 6 knees. One of 2 patients with deep infection needed femoral amputation. With correct indication and considerable surgical experience, UKA is still a good alternative, especially in the elderly patient.

Novel octaketide macrolides related to 6-deoxyerythronolide B provide evidence for iterative operation of the erythromycin polyketide synthase.

BACKGROUND: The macrolide antibiotic erythromycin A, like other complex aliphatic polyketides, is synthesised by a bacterial modular polyketide synthase (PKS). Such PKSs, in contrast to other fatty acid and polyketide synthases which work iteratively, contain a separate set or module of enzyme activities for each successive cycle of polyketide chain extension, and the number and type of modules together determine the structure of the polyketide product. Thus, the six extension modules of the erythromycin PKS (DEBS) together catalyse the production of the specific heptaketide 6-deoxyerythronolide B. RESULTS: A mutant strain of the erythromycin producer Saccharopolyspora erythraea, which accumulates the aglycone intermediate erythronolide B, was found unexpectedly to produce two novel octaketides, both 16-membered macrolides. These compounds were detectable in fermentation broths of wild-type S. erythraea, but not in a strain from which the DEBS genes had been specifically deleted. From their structures, both of these octaketides appear to be aberrant products of DEBS in which module 4 has 'stuttered', that is, has catalysed two successive cycles of chain extension. CONCLUSIONS: The isolation of novel DEBS-derived octaketides provides the first evidence that an extension module in a modular PKS has the potential to catalyse iterative rounds of chain elongation like other type I FAS and PKS systems. The factors governing the extent of such 'stuttering' remain to be determined.

[Successful therapy of an ILVEN in a 7-year-old girl with calcipotriol]. / Erfolgreiche Therapie eines ILVEN bei einem 7jährigen Mädchen mit Calcipotriol.

A 7-year-old otherwise healthy girl presented with a 2-year history of an ILVEN (inflammatory linear verrucous epidermal nevus) located on the inner part of her right upper arm. The diagnosis was histologically confirmed. Different conservative therapeutic strategies with corticosteroids, antibiotics and antimycotics produced little or no improvement. Because of encouraging reports describing the successful use of 0.005% calcipotriol ointment in patients with ILVEN, we treated our patient with this regimen. After 4 weeks we could recognize a impressive improvement and after 8 weeks the ILVEN had nearly completely disappeared. 25 weeks after withdrawal of calcipotriol, no relapse had occurred. The dramatic response to calcipotriol suggests some pathological links between ILVEN and psoriasis.

[Intracytoplasmic injection (ICSI) of cryopreserved testicular spermatozoa (Cryo-TESE): a retrospective study of the first 250 treatment cycles]. / Intrazytoplasmatische Injektion (ICSI) von kryokonservierten testikulären Spermatozoen (Kryo-TESE): Eine retrospektive Analyse der ersten 250 Behandlungszyklen.

It is reported on the results of 250 treatment cycles in which we carried out intracytoplasmic injections (ICSI) with frozen and thawed testicular spermatozoa (cryo-TESE). Up to July 1997 we treated 127 patients, 225 embryo transfers were performed (90%), and an average of 2.3 preimplantation embryos were transferred. This resulted in 53 clinical pregnancies, six patients aborted (11.3%). The pregnancy rate was 21.2% per treatment cycle, 23.5% per embryo transfer, and 41.7% per patient. This so called cumulative pregnancy rate is still about to rise, because 49 out of the 72 non-pregnant patients are still in our ICSI-program. Twenty-two children are born, 2 twins and 1 triplet. All children are healthy and without any major malformations. We conclude from these results that using cryopreserved testicular sperm for ICSI is an effective and successful approach for the treatment of severe testicular insufficiency. In comparison to the use of native testicular sperm with the necessity of repetitive testicular biopsies, cryopreservation is advantageous in many concerns (e.g. logistic, organisatoric and financial) and is therefore recommended for clinical routine.

Engineering of a minimal modular polyketide synthase, and targeted alteration of the stereospecificity of polyketide chain extension.

BACKGROUND: Polyketides are a large and structurally diverse group of natural products that include antibiotics, antifungal agents and immunosuppressant compounds. Polyketides are biosynthesised in filamentous bacteria on modular polyketide synthases (PKSs) in which each cycle of chain extension requires a different 'module' of enzymatic activities. The recently proposed dimeric model for modular PKSs predicts that even a single-module PKS should be catalytically active in the absence of other PKS components. Researchers are also interested in manipulating the stereochemical outcome of polyketide chain extension using genetic engineering of domains within each module. RESULTS: We have constructed a minimal modular PKS from the erythromycin-producing PKS (DEBS) of Saccharopolyspora erythraea. The diketide synthase (DKS1-2) consists of a single chimaeric extension module, derived from the DEBS module 1 ketoacyl-ACP synthase (KS), sandwiched between a loading module and a chain-terminating thioesterase. When DKS1-2 was expressed in S. erythraea, the strain preferentially6 accumulated the diketide (2R, 3S)-2-methyl-3-hydroxy pentanoic acid. CONCLUSIONS: These results demonstrate that, as predicted, even a single-module PKS is catalytically active in the absence of other DEBS proteins. In its normal context, the ketosynthase domain KS1 is thought to generate a (2S)-2methyl-3-hydroxy intermediate by epimerising the initial product of carbon-carbon chain formation, the (2R)-2-methyl-3-ketoester. The observed formation of the alternative (2R)-methyl-3-hydroxy product catalysed by DKS1-2 provides strong support for this proposal, and indicates how targeted alteration of stereospecificity can be achieved on a modular PKS.

[ANCA-positive lupus erythematodes profundus. Successful therapy with low dosage dapsone]. / ANCA-positiver Lupus erythematodes profundus. Erfolgreiche Therapie mit niedrig dosiertem Dapson.

A 44-year-old male patient with lupus erythematosus profundus (lupus panniculitis) (LEP) of the right cheek/infraorbital region presented as a recurrent swelling. He had only anti-neutrophil-cytoplasmic antibodies (ANCA), which have not been previously described in association with LEP. There was no indication of systemic lupus erythematosus. While chloroquine produced no benefits, the skin lesions promptly cleared with dapsone. Step by step we reduced dapsone to a final dose of 50-75 mg per week. The ANCA titer decreased in parallel to the clinical improvement.