[Intralesional Cryosurgery in the Treatment of Keloids - A Differentiation of Keloid Types for the Improvement of Patient Selection]. / Keloidbehandlung mit intraläsionaler Kryochirurgie.

The overproduction of altered collagen fibers and the overexpression of Tumor Growth Factor-ß must be blocked in order to interrupt the growth process within a keloid scar. This can barely be achieved with the classical therapeutic methods. The results of keloid treatment are difficult to predict and the recurrence rate is usually over 50 %. In addition, some of the procedures used (e. g. irradiation) may induce additional health risks. Intralesional cryosurgery offers a therapeutic alternative that has been evaluated since more than a decade. Our own experience in more than one thousand keloid treatments allows a critical evaluation of the classification in those keloid types, which are recommended to be treated with the technique and those, which may not respond.

Relation of genetic phylogeny and geographical distance of tick-borne encephalitis virus in central Europe.

Tick-borne encephalitis virus (TBEV) is the most important arboviral agent causing disease of the central nervous system in central Europe. In this study, 61 TBEV E gene sequences derived from 48 isolates from the Czech Republic, and four isolates and nine TBEV strains detected in ticks from Germany, covering more than half a century from 1954 to 2009, were sequenced and subjected to phylogenetic and Bayesian phylodynamic analysis to determine the phylogeography of TBEV in central Europe. The general Eurasian continental east-to-west pattern of the spread of TBEV was confirmed at the regional level but is interlaced with spreading that arises because of local geography and anthropogenic influence. This spread is reflected by the disease pattern in the Czech Republic that has been observed since 1991. The overall evolutionary rate was estimated to be approximately 8×10(-4) substitutions per nucleotide per year. The analysis of the TBEV E genes of 11 strains isolated at one natural focus in zdár Kaplice proved for the first time that TBEV is indeed subject to local evolution.

Toxins A and B from Clostridium difficile differ with respect to enzymatic potencies, cellular substrate specificities, and surface binding to cultured cells.

Clostridium difficile toxins A and B together are responsible for the symptoms of pseudomembranous colitis. Both toxins intoxicate cultured cells by the same mechanism but they differ in cytotoxic potency, toxin B being generally 1,000 times more potent than toxin A. Don and T84 cells were used to determine differences in the intoxication process exerted by both toxins. Three main differences were identified: (a) the specific binding of radiolabeled toxins to the cell surfaces correlated with the cytotoxic potency, (b) toxin B was found to have a 100-fold higher enzymatic activity than toxin A, and (c) toxin A was found to modify an additional substrate, Rap. The relative contribution of (a) and (b) to the difference in cytotoxic potency was determined by microinjection of the toxins. The differing enzymatic activities turned out to be the main determinant of the difference in cytotoxic potency, whereas the difference in binding contributes to a lesser degree. These findings are discussed in the context of the pathophysiological role of the toxins.

Delineation of the catalytic domain of Clostridium difficile toxin B-10463 to an enzymatically active N-terminal 467 amino acid fragment.

In an attempt to directly approach the postulated toxic domain of Clostridium difficile's TcdB-10463, eight subclones of different size and locations in the N-terminal third of the toxin were generated. Expression of these toxin fragments was checked in Western blots and the enzymatic activity of the expressed proteins was analyzed by glucosylating Ras related small GTP-binding proteins. Two polypeptides of 875 aa (TcdBc1-3) and 557 aa (TcdBc1-H) glucosylated their targets Rho, Rac and Cdc42 with the same activity and specificity as the holotoxin. In comparison 516 aa (TcdBc1-N) and 467 aa (TcdBc1-A) protein fragments exhibited highly reduced activity, while Tcdc1 and TcdB2-3 (aa 1-243 and 244-890, respectively) were enzymatically inactive. Our results indicate that all structures involved in the catalysis are located at several different sites within the 557 aa fully active fragment. The shortest enzymatically still active protein covers aa 1-467 and obviously fulfils all minimal requirements for glucosylation. The data support the postulated three domain model of 'large clostridial cytotoxins'.

Biomodel-guided stereotaxy.

OBJECTIVES: To simplify the practice of stereotactic surgery by using an original method, apparatus, and solid anatomic replica for trajectory planning and to validate the method and apparatus in a laboratory and clinical trial. METHODS: The patient is marked with fiducials and scanned by using computed tomography or magnetic resonance imaging. The three-dimensional data are converted to a format acceptable to stereolithography. Stereolithography uses a laser to polymerize photosensitive resin into a solid plastic model (biomodel). Stereolithography can replicate blood vessels, soft tissue, tumor, and bone accurately (<0.8 mm). A stereotactic apparatus is referenced to fiducials replicated in the biomodel. The trajectory for the intervention is determined and saved. The apparatus is attached to the patient fiducials, and the intervention is replicated. RESULTS: Three types of apparatus (template, Brown-Roberts-Wells frame, and D'Urso frame) were tested on phantoms and patients requiring the excision/biopsy of tumors. The localization errors determined from the phantom studies were template, 0.82 mm; Brown-Roberts-Wells frame, 1.17 mm; and D'Urso frame, 0.89 mm. The surgeons reported that clinical use of the template and D'Urso frame was accurate and ergonomic. The Brown-Roberts-Wells frame was more difficult to use and somewhat inaccurate. CONCLUSION: Biomodel-guided stereotaxy has significant advantages. It is performed quickly; it is based on simple, intuitive methodology; it enhances visualization of anatomy and trajectory planning; it enhances patient understanding; it uses inexpensive equipment; it does not require rigid head fixation; and it has greater versatility than known techniques. Disadvantages are biomodel cost and a manufacturing time of 12 to 24 hours.

Ascites from a ventriculoperitoneal shunt.

An unusual case of ascites secondary to a ventriculoperitoneal shunt is presented. The ascites did not redevelop following diversion of the distal catheter into the right atrium, and no adequate explantation was found as to why the cerebrospinal fluid was not absorbed by the peritoneum.

Cerebrovascular biomodelling: a technical note.

BACKGROUND: Recently computed tomographic angiography (CTA) and MR angiography (MRA) have been used to image cerebrovascular structures. Although CTA and MRA are accurate and sensitive imaging modalities, limitations have been identified in relation to image interpretation. Stereolithographic (SL) biomodelling is a new technology that allows three-dimensional (3D) CT and MR data to be used to accurately manufacture solid plastic replicas of anatomical structures. A prospective trial of SL biomodelling in cerebrovascular surgery has been performed to investigate the feasibility and clinical utility of this new display medium. METHODS: Fifteen patients with cerebral aneurysms and 1 patient with a cerebral arteriovenous malformation (AVM) were selected. 3D CT and/or MR angiograms were acquired and 19 solid anatomical biomodels manufactured using the rapid prototyping technology of stereolithography. The biomodels were used for patient education, diagnosis, operative planning and surgical navigation. RESULTS: The biomodels replicated the CTA and MRA source data. The accuracy of one biomodel was verified by comparison with a post mortem specimen, which corresponded exactly in the x and y planes but differed by 2 mm in the z plane. The ability to closely study an overview of complex cerebrovascular anatomy from any perspective on a solid biomodel was reported to enhance the surgeon's understanding, particularly when conventional images were equivocal. Cerebrovascular biomodels were found to be useful when positioning the patient's head for surgery, for selecting the best aneurysm clip and for the simulation of clipping. Patient informed consent was anecdotally improved. Disadvantages of the technology were the cost and manufacturing time. CONCLUSIONS: Cerebrovascular biomodelling may have utility in complex cases or when the standard imaging is felt to be equivocal.

Development of a Usutu virus specific real-time reverse transcription PCR assay based on sequenced strains from Africa and Europe.

Usutu virus (USUV) has been isolated in several African and European countries mainly from mosquitoes and birds. However, previous benign and two recent severe cases of human infections point out the need of a tool for the identification of USUV in human samples. A published real-time reverse transcription (RT) PCR assay for the detection of USUV in human blood or cerebrospinal fluid does not take into account the genetic variability of USUV in different geographic regions. Therefore, this article presents a quantitative real-time RT-PCR assay based on sequences from Europe and Africa. Primers and probe were designed in conserved regions among USUV strains that differed from closely related flaviviruses. The specificity of the assay was investigated by testing 16 other flaviviruses circulating in Africa. The sensitivity was determined by testing serial dilutions of virus and RNA standard. Intra- and inter-assay coefficients of variation were evaluated by 10 reactions in a same and in different assays, respectively. The assay provides high analytical specificity for USUV and detection limits of 1.2pfu/reaction for virus dilutions in L-15 medium or human serum and 60 copies/reaction for the RNA standard. The assay needs to be evaluated in a clinical context and integrated in standard diagnosis of flaviviral diseases.

A new approach for diagnosis of bovine coronavirus using a reverse transcription recombinase polymerase amplification assay.

Bovine coronavirus (BCoV) is an economically significant cause of calf scours and winter dysentery of adult cattle, and may induce respiratory tract infections in cattle of all ages. Early diagnosis of BCoV helps to diminish its burden on the dairy and beef industry. Real-time RT-PCR assay for the detection of BCoV has been described, but it is relatively expensive, requires well-equipped laboratories and is not suitable for on-site screening. A novel assay, using reverse transcription recombinase polymerase amplification (RT-RPA), for the detection of BCoV is developed. The BCoV RT-RPA was rapid (10-20 min) and has an analytical sensitivity of 19 molecules. No cross-reactivity with other viruses causing bovine gastrointestinal and/or respiratory infections was observed. The assay performance on clinical samples was validated by testing 16 fecal and 14 nasal swab specimens and compared to real-time RT-PCR. Both assays provided comparable results. The RT-RPA assay was significantly more rapid than the real-time RT-PCR assay. The BCoV RT-RPA constitutes a suitable accurate, sensitive and rapid alternative to the common measures used for BCoV diagnosis. In addition, the use of a portable fluorescence reading device extends its application potential to use in the field and point-of-care diagnosis.

Sandfly fever virus activity in central/northern Anatolia, Turkey: first report of Toscana virus infections.

Sandfly fever viruses (SFVs) cause febrile diseases as well as aseptic meningitis/encephalitis and include serotypes sandfly fever Sicilian virus (SFSV), sandfly fever Naples virus (SFNV) and Toscana virus (TOSV). Infections are endemic in the Mediterranean basin and data on SFV activity in Turkey are limited. In this study, sera from 1533 blood donors from the Ankara, Konya, Eskisehir and Zonguldak provinces of Turkey were evaluated for SFV exposure by indirect immunofluorescence test (IIFT) and confirmed by virus neutralization test (VNT). One hundred and two patients with central nervous system (CNS) infections of unknown aetiology were also tested via IIFT and real-time reverse-transcription PCR for SFV/TOSV. Rate of overall IgG reactivity in IIFT was 32.9% (505/1533) among blood donors. TOSV exposure was confirmed by VNT in all study regions. Exposure to the recently-identified serotype sandfly fever Turkish virus, as evaluated by VNT, was revealed in Konya and Ankara. SFNV exposure was identified in Konya and SFSV was observed to be present in all regions except Zonguldak. TOSV RNA was detected in 15.7% (16/102) and was accompanied by TOSV IgM in 25% (4/16) of the patients. Partial L and S sequences suggested that TOSV circulating in Turkey can be grouped into TOSV genotype A strains. Exposure to TOSV and other SFV serotypes was revealed in blood donors and CNS infections by TOSV were identified for the first time in Turkey. Infections are observed to be endemic in central Anatolia and should be considered as aetiologic agents in cases/outbreaks of fever and meningoencephalitis.

Trigeminal neuralgia. Surgical treatment by microvascular decompression of the trigeminal nerve root.

The surgical management of trigeminal neuralgia has always been less than ideal. Evidence accumulated over the last 10 years suggests that the condition is caused by vascular compression of the trigeminal nerve root at the brain stem. Removal of this compression by means of micro-neurosurgical techniques has so far been very effective in relieving the pain of trigeminal neuralgia while maintaining normal facial sensation. A series of 10 patients who underwent this procedure is presented. All patients lost their pain, and during the follow-up period of six to 18 months, there have been no recurrences. This experience further demonstrates that the microvascular decompression is a very useful procedure in selected patients.

Surgical relief of post-herpetic pain by excision of scarred skin.

This paper deals with the problems of post-herpetic pain. The various modes of treatment which have been tried for its relief are discussed as is our experience of excision of the scarred area of skin. The procedure has been found to offer worthwhile pain relief to a significant number of patients.

[Distribution of methadone in blood, cerebrospinal fluid, urine and organ tissues]. / Verteilung von Methadon in Blut, Liquor, Urin und Organgewebe.

This study investigated the deposition of methadone in blood, liquor, urine and organ tissues, obtained from sheep after daily administration of methadone during 23 days. One sheep was killed two hours after methadone application, one sheep after 24 hours without methadone, and one sheep after seven days without methadone. The methadone determination was performed by radioimmunoassay. The concentrations measured represent the sum of methadone and its metabolites expressed in methadone equivalent/ml or g, respectively. The highest methadone levels were found in blood and liquor in the first 30 minutes. After seven days without methadone in blood, liquor and urine was methadone not more found. Only in the synovial fluid and vitreous humor measurable methadone levels were present.

The intrinsic ability of ribosomes to bind to endoplasmic reticulum membranes is regulated by signal recognition particle and nascent-polypeptide-associated complex.

Signal peptides direct the cotranslational targeting of nascent polypeptides to the endoplasmic reticulum (ER). It is currently believed that the signal recognition particle (SRP) mediates this targeting by first binding to signal peptides and then by directing the ribosome/nascent chain/SRP complex to the SRP receptor at the ER. We show that ribosomes can mediate targeting by directly binding to translocation sites. When purified away from cytosolic factors, including SRP and nascent-polypeptide-associated complex (NAC), in vitro assembled translation intermediates representing ribosome/nascent-chain complexes efficiently bound to microsomal membranes, and their nascent polypeptides could subsequently be efficiently translocated. Because removal of cytosolic factors from the ribosome/nascent-chain complexes also resulted in mistargeting of signalless nascent polypeptides, we previously investigated whether readdition of cytosolic factors, such as NAC and SRP, could restore fidelity to targeting. Without SRP, NAC prevented all nascent-chain-containing ribosomes from binding to the ER membrane. Furthermore, SRP prevented NAC from blocking ribosome-membrane association only when the nascent polypeptide contained a signal. Thus, NAC is a global ribosome-binding prevention factor regulated in activity by signal-peptide-directed SRP binding. A model presents ribosomes as the targeting vectors for delivering nascent polypeptides to translocation sites. In conjunction with signal peptides, SRP and NAC contribute to this specificity of ribosomal function by regulating exposure of a ribosomal membrane attachment site that binds to receptors in the ER membrane.

The culture of youth and drug abuse in some European countries.

Old patterns of education are being replaced in some European countries by the different ideological norms of the various groups composing modern society. Youth has lost the sure feeling to belong to a certain culture. Drug abuse primarily represents an attempt to get over this insecurity. In an inquiry in Swiss military schools, recruits were questioned about drug and alcohol use and cigarette smoking, and this was related to the home situation. Juveniles who do not have enough affective support in the primary group seem to be more subject to the conflicts of norms going on in our society. Three general factors seem to be responsible for the drug use among juveniles: (1) the family situation of childhood, (2) the conflicts of norms of the society in which they are involved, and (3) the milieu provocation.

Transcription analysis of the genes tcdA-E of the pathogenicity locus of Clostridium difficile.

To analyse the transcription pattern of the five tcdA-E genes of the pathogenicity locus (PaLoc) of Clostridium difficile a protocol was established to purify RNA from strain VPI10463. Transcription analysis of the five tcdA-E genes showed that they were all transcribed. In the early exponential phase, a high level of tcdC and low levels of tcdA,B,D,E transcripts were detectable; this was inverted in the stationary phase, suggesting that TcdC might have a negative influence on transcription of the other genes. Three transcription initiation sites, one for tcdA and two for tcdB were determined by primer extension analysis. Readthrough transcripts from outside the locus were not obtainable, so that parts of the transcription of tcdD, tcdB, tcdA and tcdC must occur by monocistronic transcription. Within the locus all possible intergenic readthrough transcripts were detectable except that between tcdC and tcdA, a stretch of DNA interrupted by a functional transcription terminator. Thus we found mono- and polycistronic transcription of tcdA and tcdB to occur which should lead to production of a surplus of tcdA over tcdB transcripts. This would explain the surplus of TcdA over TcdB expression observed in vitro. Due to its basic nature and similarity to BcnA of Clostridium perfringens and to Orf-22 of Clostridium botulinum, TcdD is most probably a regulatory protein with DNA-binding properties. On the basis of the presented study we discuss a model for the growth-phase-related, coordinate regulation of toxin expression wherein tcdC has a negative and tcdD a positive regulatory function on transcription of the tcdD,B,E and tcdA genes.

Management of solitary melanoma metastasis of the brain.

A retrospective analysis of 31 patients operated upon for cerebral secondary melanoma was conducted. There was no operative mortality and no operative complications in 76% of cases. Significant and life-threatening complications occurred in five patients (17%). The major benefit from surgical excision is relief of symptoms: 64% had complete remission of symptoms while a further 20% were substantially improved. A few patients' lives were prolonged by surgery and there are a number of long-term survivors. Surgical excision should be performed when a patient has an accessible solitary cerebral secondary without evidence of melanoma elsewhere.