Morphology, ecology, and molecular biology of a new species of giant larvacean in the eastern North Pacific: Bathochordaeus mcnutti sp. nov.

Bathochordaeus mcnutti sp. nov. is described from the mesopelagic northeast Pacific Ocean (Monterey Bay, California, USA). Larvaceans in the genus Bathochordaeus are large, often abundant zooplankters found throughout much of the world ocean, but until recently it was unclear whether more than a single species of Bathochordaeus existed. Using remotely operated vehicles, we have made hundreds of in situ observations, compiled two decades of time-series data, and carefully collected enough specimens to determine that three species of Bathochordaeus occur in Monterey Bay: B. charon (Chun), B. stygius (Garstang), and B. mcnutti sp. nov. Bathochordaeus mcnutti is readily distinguished from its two congeners by the distinct blue outline visible around the periphery of its tail, and by other aspects of its morphology, ecology, and genetics. The abundance of larvaceans means they are ecologically important as particle processors. Species within the genus, Bathochordaeus, comprise the largest of described larvaceans.

Mouse models of genomic syndromes as tools for understanding the basis of complex traits: an example with the smith-magenis and the potocki-lupski syndromes.

Each human's genome is distinguished by extra and missing DNA that can be "benign" or powerfully impact everything from development to disease. In the case of genomic disorders DNA rearrangements, such as deletions or duplications, correlate with a clinical specific phenotype. The clinical presentations of genomic disorders were thought to result from altered gene copy number of physically linked dosage sensitive genes. Genomic disorders are frequent diseases (~1 per 1,000 births). Smith-Magenis syndrome (SMS) and Potocki-Lupski syndrome (PTLS) are genomic disorders, associated with a deletion and a duplication, of 3.7 Mb respectively, within chromosome 17 band p11.2. This region includes 23 genes. Both syndromes have complex and distinctive phenotypes including multiple congenital and neurobehavioral abnormalities. Human chromosome 17p11.2 is syntenic to the 32-34 cM region of murine chromosome 11. The number and order of the genes are highly conserved. In this review, we will exemplify how genomic disorders can be modeled in mice and the advantages that such models can give in the study of genomic disorders in particular and gene copy number variation (CNV) in general. The contributions of the SMS and PTLS animal models in several aspects ranging from more specific ones, as the definition of the clinical aspects of the human clinical spectrum, the identification of dosage sensitive genes related to the human syndromes, to the more general contributions as the definition of genetic locus impacting obesity and behavior and the elucidation of general mechanisms related to the pathogenesis of gene CNV are discussed.

[Liposuction for "body contouring" in gynecology]. / Der Einsatz der Liposuktion zum "body contouring" in der Gynäkologie.

OBJECTIVES: The aim of this study was to describe the possible use of the ultrasound-assisted liposuction and liposuction with the tumescent technique for the contouring and remodelling of superficial fat areas of women in the field of gynaecology. PATIENTS AND METHODS: Between 1997 and 1999 85 healthy female patients underwent a liposuction in the department of gynaecology of the university of Essen. The patients were divided into two groups. Thirty patients (group 1) underwent an ultrasound-assisted liposuction whereas the remaining 55 patients (group 2) were operated using only the tumescent technique. RESULTS: From the operated 582 body areas a large volume liposuction with the aspiration of more than 1,000 cc fat was performed in 48.2% of the cases. In the remaining 51.8% of the cases aspiration volumes between 300 and 1,000 cc fat were obtained. No statistically significant differences could be observed when comparing the aspirat volumes between both treatment groups (p > 0.05). Serious complications were not observed. DISCUSSION: Our data could show, that liposuction is an extremely safe method for eliminating surperficial fat depots in the sense of body contouring in gynaecology, but that it should not be used for the reduction of obese body volumes. If ultrasound-assisted liposuction is really superior to liposuction with the tumescent technique remaining uncertain, no time gain could be observed due to this technique.

Long-term follow-up of female patients with congenital adrenal hyperplasia from 21-hydroxylase deficiency, with special emphasis on the results of vaginoplasty.

OBJECTIVE: To assess, in a long-term follow-up, female patients with congenital adrenal hyperplasia (CAH), with special emphasis on vaginal functional outcome and sexual activity after vaginoplasty. PATIENTS AND METHODS: Twenty-seven patients with CAH (aged 14-33 years; six Prader grade II, 14 grade III, six grade IV and one grade V) underwent surgery between 1972 and 1988. Three of the patients underwent clitoridectomy, 24 clitoroplasty and 25 vaginoplasty (24 with a Fortunoff flap and one a 'pull-through' procedure). In 20 patients the vaginoplasty was a one-stage procedure, undertaken at a mean (range) age of 3.6 (1-9) years, and in five patients a two-stage operation. The analysis was based on the patients' history and examination; the patients also completed a questionnaire, including a psychological profile. RESULTS: Nine of the 25 patients (36%) who underwent vaginoplasty developed intravaginal stenosis; of these nine, six were Prader grade III and three grade IV. All had undergone a single-stage procedure at a mean (range) age of 4.7 (2-9) years. Of the 16 patients who answered the questionnaire, 14 had problems with their overall body image; patients in whom vaginal stenosis was corrected were particularly anxious about sexual intercourse and had problems with orgasm. CONCLUSION: The main problem during the long-term follow-up was intravaginal stenosis; all the affected patients had undergone a single-stage procedure early in life to correct ambiguous genitalia. This high rate of vaginal stenosis suggests that vaginoplasty should be undertaken at the beginning of puberty, because higher oestrogen levels may prevent stenosis and, if necessary, dilatation can be performed by the patient. These data also underscore the importance of psychological support in the treatment of children with CAH.

Isolation of a derivative of Escherichia coli-Enterococcus faecalis shuttle vector pAM401 temperature sensitive for maintenance in E. faecalis and its use in evaluating the mechanism of pAD1 par-dependent plasmid stabilization.

A derivative of the Escherichia coli-Enterococcus faecalis shuttle vector pAM401 was isolated by mutagenesis in an E. coli mutator strain. This plasmid, designated pAM401ts, was more than an order of magnitude less stable at 38 degreesC than at 30 degreesC in the E. faecalis host strain JH2-2. The E. faecalis plasmid pAD1-encoded par stability locus was cloned onto pAM401ts, and its effects on plasmid stability and host cell viability were assessed. It was found that par stabilized pAM401ts at 38 degreesC but also caused a substantial drop in cell viability three to four generations after a temperature shift from 30 to 38 degreesC. After a maximum viability drop of 94%, culture growth recovered as plasmid-free cells began to accumulate. Provision of excess RNAII, the putative par antidote, in trans attenuated cell killing. These characteristics support a postsegregational killing mechanism for par-mediated plasmid stabilization.

Clonidine prolongs fentanyl-induced ventilatory depression.

The aim of the present study was to investigate the effect of intravenously (i.v.) administered fentanyl and clonidine on ventilation in 12 healthy male volunteers (age 30.8 +/- 4.9 years) who either received fentanyl alone (1.5 > or = micrograms kg-1) or fentanyl (1.5 > or = micrograms kg-1) in combination with clonidine (3 > or = micrograms kg-1). The effect on ventilation was measured with a CO2 rebreathing system. The respiratory depression caused by fentanyl disappeared 120 min after injection. The corresponding slopes were 7430 +/- 2075 mL kPa-1 prior to (t0) and 6263 +/- 1864 mL kPa-1 120 min post-application (base-line vs. t120; P = 0.106). An impaired ventilatory response was observed during CO2 rebreathing at t120 after the injection of fentanyl and clonidine. Before drug administration, the slope of the response curves was 7700 +/- 2800 mL kPa-1, which was reduced to 5480 +/- 2135 mL kPa-1 (P < 0.035) at t120. These data suggest a prolongation of a fentanyl-induced ventilatory depression when used in combination with clonidine.

In vivo and in vitro phosphorylation of the alpha 7/PRS1 subunit of Saccharomyces cerevisiae 20 S proteasome: in vitro phosphorylation by protein kinase CK2 is absolutely dependent on polylysine.

In this paper, we show that the Saccharomyces cerevisiae 20 S proteasome subunit 1 (PRS1), recently renamed as alpha 7, is the main in vivo phosphorylated and in vitro CK2-phosphorylatable proteasome component. In vitro phosphorylation occurs only in the presence of polylysine, a characteristic that distinguishes the yeast proteasome from mammalian ones which are phosphorylated by CK2 in the absence of polylysine. A peptide reproducing the long acidic C-terminal tail of alpha 7/PRS1, where consensus CK2 phosphorylation sites are located, was also phosphorylated by the CK2 holoenzyme in a polylysine-dependent manner, suggesting that this region contains structural features responsible for this particular behavior.

Purification and characterization of protein kinase CK2 from Candida albicans: evidence for the presence of two distinct regulatory subunits beta and beta'.

Protein kinase CK2 of Candida albicans has been purified to near homogeneity by a procedure which involves chromatography on DEAE-cellulose, phosphocellulose, Q-Sepharose, and heparin-agarose. The purified enzyme has the characteristic properties of animal and yeast CK2, i.e., it utilizes ATP as well as GTP as phosphate donor, phosphorylates serine and threonine residues on casein, is inhibited by low concentrations of heparin, and is stimulated by NaCl and polycationic compounds such as polylysine, spermine, and spermidine. The native form of the enzyme exhibits a molecular mass of 159 kDa, and SDS-PAGE analysis indicates that it is composed of four polypeptides with relative molecular masses of 44, 39, 37 and 36 kDa. The 39- and 37-kDa polypeptides were identified as distinct catalytic subunits alpha and alpha' on the basis of in situ phosphorylation assays and immunological recognition with heterologous antibodies. The purified kinase undergoes autophosphorylation on the 44- and 36-kDa polypeptides, a characteristic of the beta subunits from other species. Antibodies raised against the beta subunit of Drosophila melanogaster and human CK2 crossreact only with the 36-kDa polypeptide. The 44-kDa polypeptide was identified as an unusually large beta' subunit by Western blotting with an antibody raised against the beta' subunit of Saccharomyces cerevisiae. All these data suggest that C. albicans CK2 has an alpha alpha' beta beta' heterotetrameric composition similar to that found in S. cerevisiae.

Cytokines and impaired CD8+ CTL activity among elderly persons and the enhancing effect of IL-12.

We have previously demonstrated that about 70% of elderly persons exhibit deficient cytotoxic T lymphocyte (CD8+ CTL) responses against influenza viruses when compared to young persons. Since IFN-gamma, a Th1 cytokine and IL-4, a Th2 cytokine, stimulate and inhibit CD8+ CTL responses respectively, their role(s) in the age-related CTL deficiency was investigated. Lymphocytes from young adults (34 +/- 5 years old) and elderly subjects (71 +/- 1 years old) were stimulated in vitro with influenza A/H3N2, A/H1N1 or influenza B virus for 6-7 days. The CD8+ CTL activity against virus-infected autologous target cells was significantly lower among the elderly than the young subjects (P < 0.01). Following stimulation with influenza virus, IL-4 production in both age groups was similar on day 3 but significantly higher among elderly persons on day 6 (P < 0.05). In contrast, T cells from the elderly produced significantly lower IFN-gamma than did those from young persons on both days (P < 0.05). Treatment of T cells from young and elderly adults with recombinant human IL-12, a pivotal cytokine that stimulates Th1 cytokines, resulted in enhancement of CD8+ CTL activity and IFN-gamma production in a dose dependent manner (P < 0.01). IL-12-dependent enhancement of CTL activity was not always abrogated by anti-IFN-gamma antibody treatment. These results suggest that deficient influenza virus-specific CTL activity among the elderly is attributable to a Th1 to Th2 cytokine production switch. Immunotherapy with IL-12 could represent a useful approach to correct the CD8+ CTL deficiency and cytokine imbalance among elderly humans.

[Prolongation of fentanyl-induced decreased arousal by clonidine]. / Untersuchungen zur Verlängerung einer fentanylinduzierten Vigilanzminderung durch Clonidin.

Both fentanyl and clonidine lead to a reduction of vigilance. We tested the influence of clonidine on a fentanyl induced change of vigilance using a cross-out concentration test (b-cross-out test). With local ethics committee approval, we conducted a randomised double-blinded cross-over study with 10 healthy male subjects (25-35 years old). Each volunteer received intravenously at two separate sessions 1.5 micrograms/kg fentanyl and 1.5 micrograms/kg fentanyl plus 3 micrograms/kg clonidine. For testing the level of vigilance a concentration test (b-cross-out) was used. The number of correctly revised marks was evaluated prior to drug administration (TI), directly after application, one hour later and two hours later. Our data underwent a multiple analysis of variance. With fentanyl two hours after application the mean increase of correctly revised marks was 12% when compared to those prior to application (p < 0.024). This means that with fentanyl a learning effect was achieved as described in the literature. With fentanyl and clonidine two hours after application, a decrease in vigilance of 7 1/4 in comparison with TI was seen (n.s.). This reduction of vigilance should be considered during the postoperative course and especially with regard to ambulant anaesthesia when using clonidine.

Discrimination between acid and alkali-labile phosphorylated residues on Immobilon: phosphorylation studies of nucleoside diphosphate kinase.

We have critically analyzed current methodologies for distinguishing histidine and serine phosphorylated residues in proteins and report a simple technique that assures a reliable discrimination. Electro-transfer of a phosphorylated enzyme to Immobilon membranes and its treatment at pH 1 and 14 in buffers containing 5% methanol allows unambiguous distinction between serine/threonine and histidine phosphorylation (O-phosphomonoesters and phosphoramide, respectively) since under these conditions only one type of residue is dephosphorylated. The addition of 5% methanol to all buffers was indispensable to deplete phosphate from membranes incubated successively under acid and basic conditions. The technique was applied to the study of nucleoside diphosphate kinase (NDP kinase) phosphorylation. In this enzyme, autophosphorylation of active site histidine is an accepted intermediate step in the catalytic phosphate transfer activity of nucleoside diphosphate kinase (NDP kinase). Nonetheless, a significant degree of autophosphorylation on other residues has been reported by several laboratories, and the hypothesis has been advanced that this nonhistidine phosphorylation may play an important role in NDP kinase cellular function, signaling the suppression of metastasis in the case of human NDP kinase A. Using this improved method, we show that human, Escherichia coli and Candida albicans NDP kinases are only autophosphorylated on histidine residues. In addition, we present evidence that the presence of phosphoserine after strong acid hydrolysis of the histidine autophosphorylated enzyme is in fact a nonenzymatic transphosphorylation from phosphohistidine due to the harsh acid treatment. This methodology was also applied to in vivo phosphorylation studies of C. albicans NDP kinase. We believe that the technique will be generally useful in histidine phosphorylation screenings.

[Muscle relaxation with atracurium in myasthenia gravis]. / Muskelrelaxation mit Atracurium bei Myasthenia gravis.

This report concerns a narcosis in a 42-year-old female patient suffering from myasthenia gravis in a state of clinical remission after thymectomia. Six months after thymectomia relaxometric control (Datex AMG) was performed while the patient underwent major colorectal surgery. We found a persisting sensitivity to relaxants during isoflurane anaesthesia supplemented with fentanyl. During 270 minutes of anaesthesia we needed only 25 mg atracurium (0.4 mg/kg bodyweight) only 57% of a theoretical normal dose. This is the result of a continuing muscular weakness in asymptomatic patients suffering from clinically reliable myasthenia gravis. Therefore, a reduced demand for muscle relaxants can be expected in these patients.

[Perioperative antibiotic prophylaxis in breast surgery: cefotiam versus clindamycin]. / Perioperative Antibiotikaprophylaxe in der Mammachirurgie: Cefotiam versus Clindamycin.

97 patients undergoing breast surgery received either 2 g cefotiam or 1200 mg clindamycin as i.v. singleshot application in the course of an open randomised comparison of parallel groups to evaluate the efficacy and tolerability of both antibiotic regimens. Both regimens proved to be well to-lerated and equally suitable for the prophylaxis of postoperative infections in breast surgery.

Candida albicans nucleoside-diphosphate kinase: purification and characterization.

Soluble nucleoside-diphosphate kinase (NDP kinase) from Candida albicans was purified to electrophoretic homogeneity and a partial sequence was determined. The enzyme was kinetically and physically characterized. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified enzyme revealed a single polypeptide of 17 kDa upon staining, by immunodetection with heterologous and homologous antibodies, and by autoradiography of the phosphorylated enzyme. Furthermore, isoelectric focusing of the purified native enzyme showed a single acidic band (pI 4.5). These results together with a native molecular mass of 98 kDa suggest a hexameric native enzyme composed of identical subunits. Like NDP kinases from other sources, the catalysis involves a phosphoenzyme intermediate that is rapidly formed upon incubation of the enzyme with ATP. The transfer of phosphate from phosphoprotein intermediate to nucleoside diphosphates is equally fast. Kinetic experiments indicated that GTP and ATP had the lowest Km compared to UTP, dTTP, and CTP. GDP acted as a preferred acceptor as assessed by kinetic measurements as well as by competition experiments. Experimental data are presented indicating the existence of a membrane-associated NDP kinase. Preliminary characterization of this enzyme suggests that cytosolic and membrane-associated NDP kinases are similar proteins.

[Perioperative preventive use of antibiotics in breast surgery: clindamycin vs. cefuroxime]. / Perioperative Antibiotikaprophylaxe in der Mammachirurgie: Clindamycin vs. Cefuroxim.

149 patients scheduled for breast surgery with an identified risk for infection received either 1.2 g Clindamycin (n = 79) or 1.5 g Cefuroxim given intravenously in a prospectively randomized fashion at induction of anaesthesia. Both drugs were well tolerated by all study patients. Postoperative fever was seen in one woman in the Clindamycin group vs. two women in the Cefuroxim group (n.s.). Serum and tissue levels measured 30 minutes after administration were within the therapeutic range for both antimicrobial substances.