[An "atraumatic" universal needle for single-shot regional anesthesia: clinical results and a 6 year trial in over 30,000 regional anesthesias]. / Eine "atraumatische" Universalkanüle für einzeitige Regionalanaesthesien. Klinische Ergebnisse nach sechsjähriger Erprobung bei über 30,000 Regionalanaesthesien.

The so-called "atraumatic" needle was developed by modification of two essential features of the Whitacre Spinal needle. The new atraumatic needle tip is universally suitable for all single-shot techniques of regional anesthesia. This is the result of a 6-year test period with 34,950 applications of 24- and 22-Gauge needles in spinal anesthesia, diagnostic lumbar puncture, peridural anesthesia, plexus anesthesia, peripheral nerve blocks with a Teflon-coated version (unipolar electrostimulation), and lumbar sympathetic and celiac plexus blocks. Postspinal headache was observed following 0.02% of punctures for anesthetic or diagnostic purposes. Transient monosymptomatic nerve damage occurred in 1 case after axillary block (0.009%). No permanent neurological sequelae were observed due to vascular, neural, or dural lesions. In comparison, 10 cases of persistent traumatic nerve damage were reported to be caused by conventional needles during the last decade. An analysis of these cases reveals some reasons for underestimating the risk of neurological sequelae after regional anesthesia. The routine clinical use of this type of atraumatic needle revealed no disadvantages with regard to efficacy of nerve blocks or training of anesthetists. Due to the extremely low incidence of postspinal headache, this needle has been used for spinal therapy and diagnostic lumbar punctures in outpatient pain therapy for 2 years. As of this time, the overall risk of outpatient lumbar puncture cannot be estimated. Our experience should encourage further controlled studies to evaluate criteria for excluding those patients unsuited for outpatient spinal anesthesia and lumbar puncture.

A (G+C)-rich motif in the aldolase C promoter functions as a constitutive transcriptional enhancer element.

The enzyme fructose-1,6-bisphosphate aldolase consists of three isozymes that are expressed in a tissue-specific manner. Using antibodies against aldolase B and C, it is shown that aldolase C is expressed in virtually all neuronal cell lines derived from the central and peripheral nervous system. Recently, experiments with transgenic mice indicated that a (G+C)-rich region of the aldolase C promoter might function as a neuron-specific control element of the rat aldolase C gene [Thomas, M., Makeh, I., Briand, P., Kahn, A. & Skala, H. (1993) Eur. J. Biochem. 218, 143-151). To functionally analyse this element, a plasmid consisting of four copies of this (G+C)-rich sequence, a TATA box, and the rabbit beta-globin gene as reporter was constructed. This plasmid was transfected into neuronal and nonneuronal cell lines and transcription was monitored by RNase protection mapping of the beta-globin mRNA. It is shown that the (G+C)-rich element of the aldolase C promoter directs transcription in neuronal as well as in nonneuronal cells. In contrast, the synapsin I promoter, used as a control for neuron-specific gene expression, directed transcription only in neuronal cells. In gel-retardation assays, two major DNA-protein complexes were detected with the (G+C)-rich element of the aldolase C promoter used as a DNA probe and nuclear extracts from brain and liver as a source for DNA-binding proteins. These DNA-proteins interactions could be impaired by a DNA probe that contained an Sp1-binding site, indicating that Sp1 or an Sp1-related factor binds to the aldolase C promoter (G+C)-rich element. This was confirmed by supershift analysis with antibodies specific for Sp1. The zinc finger transcription factor zif268/egr-1, also known to recognize a (G+C)-rich consensus site, did not, however, bind to the (G+C)-rich motif of the aldolase C promoter, nor could it stimulate transcription in transactivation assays from this control region. From these data, we conclude that the (G+C)-rich element of the aldolase C promoter functions as a constitutive transcriptional response element mediated by Sp1 and Sp1-related transcription factors.

Regulatory interactions of transcription factor YY1 with control sequences of the E6 promoter of human papillomavirus type 8.

Human papillomavirus type 8 (HPV-8) is a strictly cutaneous oncogenic virus known to induce malignant skin lesions in epidermodysplasia verruciformis patients. Our study shows that sequences surrounding transcription start sites of the HPV-8 oncogene E6 (nt 175-179) and comprising the presumable CCAAC and TATA boxes of the E6 promoter P175 contain a cluster of four motifs similar to the DNA recognition site of the multifunctional cellular transcription factor yin-yang 1 (YY1). Using DNase I footprinting and gel retardation tests it could be demonstrated that three of these motifs indeed act as YY1 binding sites. To test their functional relevance for P175 activity, engineered YY1 binding site mutants were analysed in the context of a P175 test vector using transient expression assays with human keratinocytes. YY1 turned out to exert both positive and negative effects upon the activity of the HPV-8 E6 promoter; binding of YY1 to a site upstream of the promoter's cap-position (BS1) activated transcription, whereas the downstream site (BS2) mediated repression. The second downstream YY1 binding site (BS3) seemed to play an auxiliary role, enhancing the negative effect of YY1 BS2. These observations define YY1 as an important cellular protein involved in the control of E6 oncogene expression of the skin-specific 'high risk' HPV-8 and emphasize the potential regulatory role of sequences located downstream of the transcription start site.