[New awareness for zoonoses using the example of rat bite fever : Case report and literature review]. / Neue Achtsamkeit für Zoonosen am Beispiel Rattenbissfieber : Fallbericht und Literaturübersicht.

BACKGROUND: Rat bite fever is a rare but potentially fatal bacterial zoonosis. The symptoms can be unspecific, but severe sepsis can be associated with involvement of different organs. CASE REPORT: A 27-year-old homeless man presented with fever, suspected meningitis, acute renal failure, unclear skin lesions as well as joint problems and muscular pain. Bite wounds were not detected. Meningitis could be excluded after lumbar puncture, and there was no evidence of endocarditis as the cause of the skin lesions. After 72 h, growth of Streptobacillus moniliformis in blood cultures was detected. Clinical symptoms were compatible with the diagnosis of rat bite fever. Calculated antibiosis with ampicillin sulbactam and doxycycline led to regression of the symptoms. CONCLUSION: Rat bite fever poses a diagnostic challenge due unspecific symptoms, diverse differential diagnostic options, and challenging microbiological detection. Patient history is of the utmost importance. Due to the rarity of the disease, this case report is intended to raise awareness.

[In macrohematuria, medication history also needs to be considered : Penicillin-induced immune hemolysis: a case report and review of literature]. / Bei Makrohämaturie auch an Medikamentenanamnese denken : Penicillininduzierte Immunhämolyse: ein Fallbericht mit Literaturreview.

BACKGROUND: Drug-mediated immune hemolysis is a rare but potentially life-threatening condition. Based on a case of penicillin-induced immune hemolysis, a structured literature review of case reports and studies on penicillin-mediated Drug-Induced Immune Hemolytic Anemia (DIIHA) was carried out. CASE REPORT: A 28-year-old male patient presented to the emergency department with gross hematuria and non-specific abdominal complaints. The patient had a 10-day history of respiratory infection with bacterial tonsillitis, treated orally with penicillin V on an outpatient basis. Laboratory diagnostics detected pathologically altered direct and indirect hemolysis parameters. After stopping the medication, the patient's condition could be stabilized. CONCLUSION: Diagnosis of penicillin-mediated immune hemolysis requires structured cooperation between clinic and laboratory, as clinical and serological findings may be highly variable with the risk of misdiagnosis. Due to the rarity of the disease, this case report is intended to raise awareness with respect to the triad of abrupt drop in hemoglobin levels in connection with drug therapy and in combination with a strongly positive direct Coombs test.

[Listeria endocarditis]. / Die Listerienendokarditis.

BACKGROUND: Bacterial infections caused by Listeria monocytogenes are rarely observed in routine clinical practice. Symptoms can range from comparatively unspecific to severe, septic courses of disease with cardiac, pulmonary, abdominal or bone involvement. CASE REPORT: A 74-year-old male patient with a history of mitral valve replacement and pacemaker implantation presented to the emergency department with high fever and disturbed coagulation. Transesophageal echocardiography revealed mitral valve endocarditis with early abscess formation. In blood cultures, growth of Listeria monocytogenes was detected after 20 h. Despite immediately initiated calculated antibiotic therapy and following adaption to the resistogram, the mitral valve finding was progressive and cardiosurgical intervention became necessary. CONCLUSION: Diagnosis of endocarditis caused by Listeria requires structured collaboration between cardiologists, imaging and laboratory. This case report aims to increase awareness of potentially Listeria-induced organ manifestations.

Evaluation of carcinogenic potential of 17alpha-ethinylestradiol in a host-mediated in vivo/in vitro assay system.

A host-mediated assay system for detection of the transforming activity of different chemical carcinogens on peritoneal macrophages has been previously established. Directly, as well as indirectly acting carcinogenic substances administered intraperitoneally to NMRI mice could be examined in this way. Resident macrophages were recovered by peritoneal lavage from treated and untreated mice and cultured in soft agar. After 5-6 days normal and transformed cells could be distinguished. By the use of this system an immortalized macrophage-like cell line was derived from the peritoneal cells of NMRI mice treated with 17alpha-ethinylestradiol. This cell line enabled us to perform additional investigations on the underlying molecular effects of 17alpha-ethinylestradiol, such as detection of the transformation specific polypeptides as surrogate markers for transformation. The investigation reported here describes the cell-transforming and oncogenic potential of 17alpha-ethinylestradiol.

Purification of prostaglandin D synthase by ceramic- and size exclusion chromatography.

Prostaglandin D synthase (L-PGDS) is a major glycosylated polypeptide in cerebrospinal fluid (CSF). The overexpression of L-PGDS in inflamed bovine mammary glands indicates its role as biomarker. No diagnostic tool for the quantitative detection of L-PGDS in cows has been reported. Immunometric ELISA tests might help to identify inflamed bovine tissue. The isolation of pure bovine L-PGDS, which is required for the generation of monoclonal antibodies, is an important prerequisite for a diagnostic ELISA test. Our goal was to identify a suitable technique to generate pure L-PGDS from bovine substrates. In the present study a two-step method for the purification of bovine CSF using ceramic hydroxyapatite chromatography followed by size exclusion chromatography is described. Subsequently, the identification of bovine L-PGDS was demonstrated by Western blot analysis and the high grade of the pure product was shown by 2-D PAGE. The yield of purified L-PGDS was 6.8 mg/l bovine CSF. L-PGDS from bovine CSF is shown to consist of multiple isoforms identical in molecular mass and pI values to those in previously described secretions of inflamed bovine mammary glands. In addition, the method was successfully applied to the purification of L-PGDS from human CSF.

Tumorigenic potential and the molecular mechanism of the carcinogenic effect exerted by 2-nitroanisole.

The host-mediated in vitro/in vivo assay system was used to evaluate the tumorigenic potential of the aromatic nitro compound 2-nitroanisole (2-NA). After intraperitoneal administration of the compound, resident macrophages were recovered by peritoneal lavage from treated and untreated mice and cultured in soft agar. 2-NA was shown to be carcinogenic, and the tumorigenic potential was evaluated. Additionally, by establishment of a transformed peritoneal macrophage cell line, the underlying molecular mechanism of 2-NA's carcinogenic effect was studied.

12-O-retinoylphorbol-13-acetate (RPA) behaves like a retinoid-derivative in binding to retinol-binding-protein: a correlation with its specific action as an incomplete tumor promoter?

12-O-retinoylphorbol-13-acetate (RPA), an incomplete tumor promoter of the phorbol ester type and protein kinase C (PKC) activator, consists of two characteristic structural elements: the phorbol body and the retinoyl ester chain. Therefore, possible binding of the incomplete tumor promoter RPA to the human transport protein retinol-binding-protein (RBP) has been examined by molecular modeling methods and experimental binding studies. The calculated prediction of binding properties was primarily based on a comparative geometrical approach. It was shown that the beta-ionone-ring of RPA was not altered within the binding pocket of RBP (molecular modeling) compared to retinoic acid (X-ray crystallographic data). The torsion angle C5'-C6'-C7'-C8', determining the conformation of the RBP-beta-ionone-ring relative to the isoprene tail, is rotated by 42 degrees for RPA compared to retinol and to retinoic acid, respectively. Combining all the results from force field calculations, MD simulations and geometrical comparisons, the conclusion could be drawn that RPA should be able to bind to RBP. This interaction should be less strong than that with its natural ligand retinol or with retinoic acid. This prediction was proven experimentally. RPA was able to compete with retinoic acid for binding at RBP in human plasma. The binding properties were investigated using 3H-labeled retinoic acid in homologous and heterologous competition studies in a one-dimensional native polyacrylamide gel electrophoresis system. An approximately 2000-fold weaker binding of RPA to RBP as compared to retinoic acid was determined experimentally, confirming the prediction of the molecular modeling approach. The characteristic behaviour of RPA as an incomplete promoter, due to possible binding to PKC and RBP, is discussed.

Improved purification of beta-lactoglobulin from acid whey by means of ceramic hydroxyapatite chromatography with sodium fluoride as a displacer.

The successful separation of beta-lactoglobulin from other bovine whey proteins was performed by ceramic hydroxyapatite chromatography with a fluoride ion gradient in phosphate buffer as displacement agent. The method was applied to acid whey originating from milk of healthy as well as of mastitic cows. beta-Lactoglobulin was completely eluted in one peak at a fluoride concentration of about 0.6 mol/l. The purity of beta-lactoglobulin in this fraction was at least 96% if whey from healthy milk was processed. Co-eluted contaminants are traces of immunoglobulin G, serum albumin and lactoferrin. In case of mastitic whey the proportion of beta-lactoglobulin is diminished as the amounts of immunglobulin G, serum albumin and lactoferrin are increased within this fraction. Size exclusion chromatography on Superdex 75 pg effectively removed contaminants resulting in a purity for beta-lactoglobulin from normal whey of approximately 99%. The yield of beta-lactoglobulin from physiological whey was 50-55% referring to the fraction highly enriched with beta-lactoglobulin by hydroxyapatite chromatography. In case of mastitic milk the higher amounts of contaminants were also removed successfully by size exclusion chromatography.

Identification of immunorelevant genes from greater wax moth (Galleria mellonella) by a subtractive hybridization approach.

In this study we have analyzed bacterial lipopolysaccharide (LPS) induced genes in hemocytes of the Lepidopteran species Galleria mellonella using subtractive hybridization, followed by suppressive PCR. We have found genes that show homologies to molecules, such as gloverin, peptidoglycan recognition proteins and transferrin known to be involved in immunomodulation after bacterial infection in other species. In addition, a few molecules previously not described in the innate immune reactions were detected, such as a RNA binding molecule and tyrosine hydroxylase. Furthermore, the full-length cDNA of a LPS-induced molecule with six toxin-2-like domains is described to be a promising candidate to further elucidate the relationship between toxin- and defensin-like domains in arthropod host defense.

Lipocalin-type prostaglandin D synthase in milk: a new biomarker for bovine mastitis.

The whey protein pattern of milk from animals affected by mastitic inflammation was resolved by two-dimensional gel electrophoresis (2D-PAGE) and compared to milk from unaffected cows. Inflammation caused the appearance of four spots aligned at a molecular weight level of 26 kDa and over a pH-region of 5.0 to 6.4. The spots excised from 2D gels were treated with chymotrypsin and the resulting peptides analyzed by MALDI-TOF mass spectrometry and RP-HPLC. All four spots yielded highly similar chymotryptic peptide mass fingerprints as well as chromatographic peak patterns. A database search could identify the four spots as isoforms of the bovine prostaglandin D synthase (PGD-S). In one of the isoforms a defined cysteine residue was shown to be oxidized to a sulfonic acid.

Peritoneal macrophages from 17 alpha-ethinylestradiol-treated NMRI mice secrete transformation-specific low molecular weight proteins.

Analysis of protein secretion was performed for a macrophage-like cell line, which was established from the peritoneal cells of NMRI mice treated with 17 alpha-ethinylestradiol. The protein secretion pattern was investigated by computerized analysis of high resolution two-dimensional gel electrophoresis (2-D PAGE) and compared to that of control macrophages, intraperitoneally activated by bacterial lipopolysaccharide. The transformed cells encode a number of low molecular weight proteins (10-20 kDa), which were not observed in control cells under identical experimental conditions. In conclusion the transformation of peritoneal macrophages by 17 alpha-ethinylestradiol leads to an upregulation of polypeptides, in particular of low molecular weight proteins. A high similarity between the induced low molecular weight protein secretion by macrophages of 17 alpha-ethinylestradiol-treated and that of 2,3,7,8-tetrabromodibenzo-p-dioxin-treated mice was found.

A novel polypeptide p10 expressed in tumor-promoter-treated murine epidermis and in untreated neonatal murine epidermis.

12-O-Tetradecanoyl-phorbol-13-acetate (TPA) has been shown to induce a broad variety of morphological and metabolic changes in mouse skin. These include gross changes in cell growth as well as subtle changes regarding gene expression. Changes in the induction of specific proteins have been described, but it is not yet clear, if there exists a strict causal correlation between the induction of expression of specific proteins and the phenomenon of tumor promotion. At least some of these proteins might also be correlated with an accompanying hyperproliferative effect. This has been shown for protein p10, a 10 kDa protein not found in untreated adult mouse epidermis. The expression of this protein can be induced de novo after the topical administration of TPA. It is also shown, that p10 is expressed in untreated neonatal murine epidermis. Due to the induction of this protein by irritation, hyperproliferation-causing tumor promoters and due to the observable abundance in neonatal epidermis, this protein seems not strictly related to tumor promotion, but rather to proliferation and differentiation.

[Transsexuality--a multidisciplinary problem. Which therapeutic methods proved to be successful?]. / Transsexualität--eine multidisziplinäre Problematik. Welche Therapieansätze haben sich als tauglich erwiesen?

Transsexualism as a phenomenon clearly contrasts the common understandings of sexuality. This can lead to conflicts for the transsexual himself as well as for those being confronted. Medical care and legislative offer some assistance for patients, which has been proofed as suitable and helpful. These strategies have not been developed by a theoretical approach but pragmatically and can still be further improved. To be able to receive a higher acceptance of this phenomenon in society and to achieve concrete improvement of the patient's personal situation the challenge still exists to understand mechanisms underlying the transsexual experience.

Mechanistically different acting tumor promoters induce consistent qualitative and quantitative changes in protein expression patterns of murine epidermis.

Systematic screening of different organs has shown that induction of particular proteins by 12-0-tetradecanoyl-phorbol-13-acetate is specific for mouse skin. Different compartments of skin showed all the same expression. For experimental reasons we focused the analysis performed thereafter on the epidermis. As an end- point of mouse skin tumorigenesis papilloma and carcinoma were also screened for changes in protein expression. We focused our efforts on the characterization and identification of selected, de novo expressed proteins induced by various tumor promoters in murine epidermis. The analysis presented here consists of functional characterization. Having found differences in protein expression in TPA-treated murine epidermis compared with acetone-treated control mice, we then tried to correlate the expression to the signal transduction pathway which the tumor promoter exerts its activity through. For this reason we screened protein expression patterns after topical administration of mechanistically differently acting tumor promoters, which were inflammatives not bearing tumor promoting activity anti-promoters or a combination of tumor promoters with modulators of tumor promotion. The epidermis of untreated newborn mice served as a model for high cellular proliferation and differentiation. Alterations were compared with changes observed after tumor promoter application.

A novel protein (p10) induced by 12-O-tetradecanoyl-phorbol-13-acetate (TPA) and other hyperplasiogenic tumor-promoting and non-promoting agents in murine epidermis.

Treatment of murine epidermis with the tumor promotor 12-O-tetradecanoylphorbol-13-acetate (TPA) shows characteristic and significant changes in protein expression analyzed by 2D PAGE, compared to that of acetone-treated mouse epidermis. Of the seven de novo expressed proteins in TPA treated murine epidermis, one is a 44 kDa protein (p44) located nearby actin, and six proteins are in the low-molecular range between 10-20 kDa (p10, pY, pCa, p1, p2 and p3). Interestingly, the incomplete promoting and inflammative hyperplasiogen 12-O-retinoylphorbol-13-acetate (RPA) and the non-promoting but inflammative and hyperplasiogenic calcium-ionophore A23187 induced the same pattern of proteins observed in the pidermis of mice treated with TPA, with minor quantitative differences. In all cases, p10 expression was quantitative the most abundant. Partial sequencing of this protein has led to the conclusion that it is a novel protein with no such sequences in the database comparisons using FASTA and TFASTA computer programs of Genetics Computer Group. The data presented here do not strictly support the functional role of de novo induced proteins to tumor promotion, but show a causal relationship to hyperplasiogenic potency of TPA, RPA and A23187.

A follow-up study for estimating the effectiveness of a cross-gender hormone substitution therapy on transsexual patients.

This follow-up study was carried out to validate the effectiveness of cross-gender hormone therapy embedded in a multistep treatment concept for transsexual patients. This therapy described in detail by the authors elsewhere and presented briefly below provides cross-gender hormone substitution to obtain an assimilation of secondary sex characteristics to the desired sex as quickly as possible. Personal and social background data of 46 male-to-female (M-to-F) and 42 female-to-male (F-to-M) patients passing through different stages of the treatment concept were included. In the Endocrinological Outpatient Clinic of the Max-Planck-Institute/Munich the effectiveness of cross-gender hormone replacement therapy as well as frequency and distribution of side effects were examined by follow-up examination of endocrinological parameters. Cross-gender hormones were administered either parenterally or orally. Blood samples were collected routinely after 2 to 6 months depending on the duration of hormone substitution and complication rate. The incidence of hyperprolactinemia in estrogen-treated M-to-F transsexuals lies in the range of studies published before, whereas the number of patients developing galactorrhea is significantly lower in our patients. The incidence of thromboembolic events during the time of cross-gender hormone treatment in our patients is negligible. Changes in hematological parameters are observed under cross-gender hormone therapy. With the cross-gender hormone regimen performed by us it is possible to generate less side effects in the treatment of transsexual patients than described before.

Transsexualism and osteoporosis.

The aim of this study was to investigate whether and to what extent our regime of cross-gender hormone replacement therapy might influence osteoporosis development in transsexual patients. We found that after long-term therapy the bone densities of our cross-gender hormone-treated transsexual groups (10 male-to-female and 10 female-to-male) did not show significant differences compared to those of the corresponding biological sex. Moreover, the bone-density during therapy pointed out very little variability and that independent of the gender-alteration (transsexuality-direction) and the age of the transsexuals. Our results indicate that for transsexual patients treated with cross-gender hormone replacement therapy the risk of developing osteoporosis is low.

Low molecular weight proteins secreted by peritoneal macrophages obtained from 2,3,7,8-tetrabromodibenzo-p-dioxin-treated NMRI mice.

The protein secretion patterns in a macrophage-like cell line (CBrD), established from the peritoneal cells of NMRI mice treated with the dioxin analog 2,3,7,8-tetrabromodibenzo-p-dioxin (TBrDD), were analyzed by high resolution two-dimensional gel electrophoresis (2-D PAGE), and compared to the pattern of proteins secreted by control macrophages which were intraperitoneally activated by bacterial lipopolysaccharide. The most striking alterations were observed in the low molecular range. The transformed cells encode a number of low molecular mass proteins (10-20 kDa) which were not detected in control cells under identical experimental conditions. The protein pattern with respect to isoelectric point, molecular weight, optical density (OD) and area of the spot (in mm2) has been depicted by computer analysis in relation to a standardized spot outline and the spot's background (in OD). It is concluded that the transformation of murine peritoneal macrophages by TBrDD leads to an upregulation of proteins, in particular of low-molecular-weight proteins.

Multistep treatment concept of transsexual patients.

Here we present a pragmatic multistep approach for the treatment of transsexual patients. The importance of an individually designed cross-gender hormone replacement therapy embedded in a multidisciplinary treatment concept, provided by psychiatrists, endocrinologists and surgeons, is demonstrated. Following this concept outcome of therapy has been improved in the last years. Over the last 5 years we have gained substantial experience in the cross-gender hormone treatment of transsexual patients. By continuous follow-up examinations and therapy adjustment the risk of side effects accompanying this therapy has been significantly minimized. This report is designed as a guideline to the clinical endocrinologist for the handling and treatment of transsexual patients.