[Persistent nasal obstruction following septoplasty: deviated nasal pyramid and perpendicular plate]. / Nasenatmungsbehinderung nach Septumplastik: Deviation der Lamina perpendicularis bei Schiefnase.

BACKGROUND: Functional septorhinoplasty (SRP) is a surgical procedure for improvement of symptomatic deformity of the inner and outer nose. Due to inadequate preoperative analysis and consequently insufficient surgical indication, septoplasty (SPL) alone may be performed. A frequent cause for revision septorhinoplasty (rSRP) in our institution was found to be due to a deviated bony pyramid in compound with a contralateral high deviation of the perpendicular plate, ipsilateral reluxation of the cartilaginous septum and a resulting nasal valve stenosis. MATERIALS AND METHODS: The medical records of all patients undergoing revision septorhinoplasty in our institution from 2005-2011 were evaluated retrospectively.398 patients underwent rSRP, in 57 patients (14%) a deviated bony nose after septoplasty was diagnosed. In all cases a deviated bony nose and contralateral deviation of the perpendicular plate was found and corrected by rSRP. The nasal breathing was improved in all cases. This was also shown in an increase of endonasal volume and nasal air flow in acoustic rhinometry and active anterior rhinomanometry. CONCLUSION: A deviated nasal pyramid and perpendicular plate was found to be a common cause for persistent nasal obstruction after septoplasty. A careful preoperative analysis of the nose, the pathology of the bony and cartilaginous septum and the nasal valves is mandatory in order to recognize anatomical details and set up the right indication for rhinosurgical interventions.

[Neovascularisation and free microsurgical transfer of cartilage-engineered constructs]. / Neovaskularisation und freier mikrochirurgischer Transfer von in vitro gezüchteten Knorpelkonstrukten.

BACKGROUND: Clinical imperatives for new cartilage to replace or restore the function of traumatized or missing tissue as a consequence of trauma, inherent malformations or disease has led to the need for therapies or procedures to generate cartilage for clinical applications. To ensure shape, function, and survival, in vitro cartilage-engineered constructs need to be revascularized. This study presents a viable method for neovascularization and free microsurgical transfer of these in vitro constructs. MATERIAL AND METHODS: Twelve female Chinchilla Bastard rabbits were operated. Cartilage-engineered constructs were created by isolating chondrocytes from auricular biopsies, amplifying in monolayer culture, and then seeding them onto polycaprolactone scaffolds. In each prefabricated skin flap, three in vitro cartilage-engineered constructs measuring 2×2×0.5 cm and one construct without cells, which served as the control, were implanted beneath an 8×15-cm random-pattern skin flap, neovascularized by implantation of an arteriovenous vascular pedicle with maximal blood flow. After 6 weeks, the neovascularized flaps with embedded cartilage-engineered constructs were completely removed based on the newly implanted vascular pedicle, and then freely retransferred into position using microsurgery. Macroscopic observation, histology, selective microangiography, and immunohistochemistry were performed to determine the construct vitality, neovascularization, and new cartilage formation. RESULTS: All neovascularized skin flaps with embedded tissue-engineered cartilage constructs were effectively free-transferred as free flaps. The implanted constructs were protected and well integrated within the flap. All constructs were well neovascularized and showed histologically stability in both form and size. Immunohistology showed the existence of cartilage-like tissue with extracellular matrix neosynthesis. CONCLUSION: Our experimental study revealed the reliable ability of neovascularization and free microsurgical transplantation of cartilage-engineered constructs using prefabricated flaps. With respect to effective clinical application, engineered cartilage composed of a patient's own cells can become a feasible option for the reconstruction of large cartilage defects or auricular reconstruction using this method. The procedure also represents a promising alternative for clinical practice due to minimal donor site morbidity and favorable aesthetic outcomes.

[Microsurgical thumb replantation following traumatic amputation: preliminary results in Hanoi, Vietnam]. / Daumenreplantation nach traumatischer Amputation : Erste Ergebnisse in Hanoi, Vietnam.

INTRODUCTION: Replantation using microsurgical techniques is a fairly new procedure in Vietnam. We reviewed and evaluated our 7-year results of thumb replantation in Vietnamese patients following traumatic thumb amputation. MATERIAL AND METHODS: Traumatic thumb amputations between September 1999 and July 2006 were reexamined and evaluated. Postoperative functional results were evaluated based on four main criteria: 1) the patient's subjective attitude regarding the replanted thumb, 2) the degree of mobility of the replanted thumb compared with its counterpart, 3) the level of sensitivity of the replanted thumb, and 4) the objective ability to perform daily tasks. RESULTS: A total of 26 patients were documented. The duration of follow-up averaged 22 months (range 6-72 months). The success rate of replantation was 81%. A rating of either "good" or "very good" was obtained for 81% of the replanted thumbs. DISCUSSION: Vascular thrombosis was the cause of all failures. Proper debridement, standardized microvascular techniques, timely detection of thrombosis formation, and reoperation using vein grafts play a decisive role in the final success.

Diagnosis and monitoring of murine histoplasmosis by a nested PCR assay.

A newly developed nested PCR assay was applied to murine models of histoplasmosis. ICR and BALB/c mice were intravenously infected with Histoplasma capsulatum and sacrificed up to 29 days later. Samples of blood, spleen, and lung homogenates were cultured and examined by the PCR assay. In the ICR mouse model, 265 of 319 organ samples showed concordant results. With 7 samples, the culture was positive and the PCR assay was negative whereas a positive PCR but a negative culture were obtained with 47 samples (P < 0.0001 according to McNemar's test). Organ homogenates and blood samples of either spontaneously cured or treated BALB/c mice were PCR negative. The nested PCR assay performs excellently in the monitoring of spontaneously and treatment-cured murine histoplasmosis. It limits the infection risks of the laboratory staff and might be of diagnostic value for humans.

Cytological and biochemical characterization of the FtsA cell division protein of Bacillus subtilis.

The actin-like protein FtsA is present in many eubacteria, and genetic experiments have shown that it plays an important, sometimes essential, role in cell division. Here, we show that Bacillus subtilis FtsA is targeted to division sites in both vegetative and sporulating cells. As in other organisms FtsA is probably recruited immediately after FtsZ. In sporulating cells of B. subtilis FtsZ is recruited to potential division sites at both poles of the cell, but asymmetric division occurs at only one pole. We have now found that FtsA is recruited to only one cell pole, suggesting that it may play an important role in the generation of asymmetry in this system. FtsA is present in much higher quantities in B. subtilis than in Escherichia coli, with approximately one molecule of FtsA for five of FtsZ. This means that there is sufficient FtsA to form a complete circumferential ring at the division site. Therefore, FtsA may have a direct structural role in cell division. We have purified FtsA and shown that it behaves as a dimer and that it has both ATP-binding and ATP-hydrolysis activities. This suggests that ATP hydrolysis by FtsA is required, together with GTP hydrolysis by FtsZ, for cell division in B. subtilis (and possibly in most eubacteria).

Improved plasmid vectors for the production of multiple fluorescent protein fusions in Bacillus subtilis.

The intrinsically fluorescent green fluorescent protein has been used in many laboratories as a cytological marker to monitor protein localisation in live cells. Multiple spectrally modified mutant versions and novel fluorescent proteins from other species have subsequently been reported and used for labelling cells with multiple fluorescent protein fusions. In this work we report the design and use of vectors containing some of these spectral variants of GFP for use in the Gram positive bacterium Bacillus subtilis. These vectors complement those previously described (Lewis and Marston, 1999. Gene 227, 101-109) to provide a large suite of plasmid vectors for use in this and other related Gram positive organisms. Using these vectors we have been able to directly demonstrate the sequential assembly/disassembly of proteins involved in the generation of cellular asymmetry during development.

Direct interaction between the cell division protein FtsZ and the cell differentiation protein SpoIIE.

SpoIIE is a bifunctional protein with two critical roles in the establishment of cell fate in Bacillus subtilis. First, SpoIIE is needed for the normal formation of the asymmetrically positioned septum that forms early in sporulation and separates the mother cell from the prespore compartment. Secondly, SpoIIE is essential for the activation of the first compartment-specific transcription factor sigma(F) in the prespore. After initiation of sporulation, SpoIIE localizes to the potential asymmetric cell division sites near one or both cell poles. Localization of SpoIIE was shown to be dependent on the essential cell division protein FtsZ. To understand how SpoIIE is targeted to the asymmetric septum we have now analysed its interaction with FtsZ in vitro. Using the yeast two-hybrid system and purified FtsZ, and full-length and truncated SpoIIE proteins, we demonstrate that the two proteins interact directly and that domain II and possibly domain I of SpoIIE are required for the interaction. Moreover, we show that SpoIIE interacts with itself and suggest that this self-interaction plays a role in assembly of SpoIIE into the division machinery.

Characterization of a morphological checkpoint coupling cell-specific transcription to septation in Bacillus subtilis.

Early in the process of spore formation in Bacillus subtilis, asymmetric cell division produces a large mother cell and a much smaller prespore. Differentiation of the prespore is initiated by activation of an RNA polymerase sigma factor, sigmaF, specifically in that cell. sigmaF is controlled by a regulatory cascade involving an anti-sigma factor, SpoIIAB, an anti-anti-sigma factor, SpoIIAA, and a membrane-bound phosphatase, SpoIIE, which converts the inactive, phosphorylated form of SpoIIAA back to the active form. SpoIIE is required for proper asymmetric division and much of the protein is sequestered into the prespore during septation. Importantly, activation of sigmaF is dependent on formation of the asymmetric septum. We have now characterized this morphological checkpoint in detail, using strains affected in cell division and/or spoIIE function. Surprisingly, we found that significant dephosphorylation of SpoIIAA occurred even in the absence of septation. This shows that the SpoIIE phosphatase is at least partially active independent of the morphological event and also that cells can tolerate significant levels of unphosphorylated SpoIIAA without activating sigmaF. We also describe a spoIIE mutant in which the checkpoint is bypassed, probably by an increase in the dephosphorylation of SpoIIAA. Taken together, the results support the idea that sequestration of SpoIIE protein into the prespore plays an important role in the control of sigmaF activation and in coupling this activation to septation.

Partial respecification of nasotemporal polarity in double-temporal chick and chimeric chick-quail eyes.

In chick embryos, naso-temporal polarity of the retina becomes established before Hamburger-Hamilton stage 10. To examine the plasticity of the early eye anlage, double-temporal eyes were made using stage 10-11 (E1.5) chick embryos and stage 8-9 quail embryos. In vivo and in vitro experiments revealed that these double-temporal compound eyes were not completely temporal but nasal in a large peripheral part of the graft. Four hours after transplantation, the nasal-specific fork head transcription factor CBF1 was not expressed in double-temporal eyes but was clearly detectable 24 h later. This suggests that in the peripheral part of the graft, temporal positional values were changed into nasal positional values by a respecification process.

Prespore-specific gene expression in Bacillus subtilis is driven by sequestration of SpoIIE phosphatase to the prespore side of the asymmetric septum.

The spoIIE gene is essential for the compartment-specific activation of transcription factor sigmaF during sporulation in Bacillus subtilis. SpoIIE is a membrane protein that is targeted to the potential sites of asymmetric septation near each pole of the sporulating cell. The cytoplasmic carboxy-terminal domain of SpoIIE contains a serine phosphatase that triggers the release of sigmaF in the prespore compartment after septation. To understand how septum-located SpoIIE is activated selectively in the prespore, we examined the distribution of a SpoIIE-GFP fusion protein. We show that the polar bands of SpoIIE protein actually form sequentially and that the most prominent band develops at the pole where the prespore forms. We also show that the protein is sequestered to the prespore side of the asymmetric septum. Sequestration of SpoIIE into the prespore compartment provides a mechanism that could explain the cell specificity of sigmaF activation.

Bifunctional protein required for asymmetric cell division and cell-specific transcription in Bacillus subtilis.

During sporulation in Bacillus subtilis an asymmetric cell division gives rise to unequal progeny called the prepore and the mother cell. Gene expression in the prespore is initiated by cell-specific activation of the transcription factor sigma(F). Three proteins participate in the regulation of sigma(F) activity. The first, SpoIIAB, is an inhibitor of sigma(F), that is, an anti-sigma factor. SpoIIAB is also a protein kinase that catalyzes phosphorylation of the second regulatory protein SpoIIAA (the anti-anti-sigma factor), and thus inactivates it. A third protein, SpoIIE, was shown recently to be able to dephosphorylate SpoIIAA-P in vitro. Here we show that SpoIIE is a bifunctional protein with two critical roles in the establishment of cell fate. First, we confirm by the use of in vivo experiments that it regulates the release of sigma(F) activity by dephosphorylating SpoIIAA-P. Second, we show that SpoIIE is needed for normal formation of the asymmetric septum that separates the prespore from the mother cell. Combination of these two functions in a single polypeptide may serve to couple the release of the cell-specific transcription factors with the formation of the differentiating cells.

Control of the cell-specificity of sigma F activity in Bacillus subtilis.

Sporulation in Bacillus subtilis is a simple developmental system involving the differentiation of two cell types that are formed by an asymmetric cell division. Major changes in the pattern of transcription during sporulation are brought about by the synthesis of new sigma factors (sigma), which are subunits of RNA polymerase that determine promoter specificity. Transcription in the smaller prespore cell type is initiated by a sigma factor called sigma F, the activity of which is subject to tight spatial and temporal control. It is negatively regulated by an anti-sigma factor, SpoIIAB, which is in turn controlled by an anti-anti-sigma factor, SpoIIAA. SpoIIAA and SpoIIAB participate in two contrasting reactions in vitro. In the presence of ATP, the proteins interact transiently and SpoIIAA is inactivated by phosphorylation on a specific serine residue; SpoIIAA then remains free to inhibit sigma F. In the presence of ADP, SpoIIAA binds tightly to SpoIIAB, and sigma F is set free. Release of sigma F activity in vivo might thus be effected by a prespore-specific reduction in the ATP/ADP ratio. Genetic experiments have implicated a fourth protein, called SpoIIE, in this system. It now appears that SpoIIE has two important and independent functions in the establishment of the prespore-specific transcription by sigma F. First it regulates sigma F activity, probably acting as a phosphatase to regenerate the active, non-phosphorylated form of SpoIIAA. Second it controls the formation of the septum that generates the prespore compartment. Combination of these two functions in a single polypeptide may provide a means of coupling gene expression with morphogenesis.

[A comparative study of the efficacy and tolerance of dipotassium clorazepate and flunitrazepam for oral premedication]. / Vergleichende Untersuchung der Wirksamkeit und Verträglichkeit von Dikaliumchlorazepat und Flunitrazepam zur oralen Prämedikation.

The literature shows that benzodiazepines, in view of their anxiolytic, sedative, amnesic, muscle relaxant and anticonvulsive action, are the most important substances for premedication. Eminent workers regard anxiolysis as the most important aim of premedication. In the present clinical study, oral administration of the two different benzodiazepine derivatives, flunitrazepam (F) and chlorazepate dipotassium (CD) have been explored with a view to side effects, tolerance, quality of sleep during the night, anxiolytic effect and sedation. The study involved 108 women patients aged from 20 to 60 years (ASA class I or II), all scheduled to undergo gynecological surgery in general anesthesia. There were also 20 women who received no premedication. The three groups of patients were further divided into early (operation started before 10:30 a.m.) and late-operation (operation started after 10:30 a.m.) groups. The test drugs were administered as follows: 43 women received 50 mg CD p.o. on the evening before the operation, followed by 25 mg p.o. in the morning; 45 women received 2 mg F p.o. on the evening before the operation, followed by 1 mg p.o. in the morning. All patients took the preoperative premedication at 7 o'clock in the morning. Following this medication, the anxiolytic, sedative, and amnesic effects, side effects, vigilance and O2 saturation (SaO2) were determined at defined points during the day of the operation and the 1st postoperative day. Blood pressure and heart rate were recorded and interpreted as physiological stress parameters. Anxiolysis was determined using the Erlangen Anxiety Scale (EAS) of Galster and Spörl; the degree of sedation was assessed by the anesthesiologist; amnesia was determined by the patients' recognition of picture cards; vigilance and side-effects were assessed by standardized questionnaires. Both active drugs clearly improved the quality of sleep in the night before the operation over that experienced with no premedication. There were no significant differences among the three groups in the physiological stress parameters. The preoperative SaO2 saturation was decreased significantly by oral F, but it was always more than 95%. CD had little influence on the SaO2. Unwanted somatic symptoms were found a little more frequently in the group without any premedication. There were no signs of restricted tolerance for either of the test drugs. In the premedicated groups, pre- and postoperative anxiety decreased significantly.(ABSTRACT TRUNCATED AT 400 WORDS)

Pore formation associated with the tail-tip protein pb2 of bacteriophage T5.

Upon binding of bacteriophage T5 tails to purified FhuA receptor protein the tail-tip protein pb2 became extremely sensitive to trypsin and other proteases. However, when T5 tails were bound to FhuA integrated into liposomes, pb2 was found to retain some resistance to trypsin. Electron microscopic examination of tail-liposome complexes supported the idea that trypsin resistance of pb2 in such complexes was caused by insertion of the tail-tip into the liposomes. pb2 was isolated from tails by treatment with sodium dodecyl sulfate and was further purified by gel filtration using a fast protein liquid chromatography system. pb2 obtained with this procedure was most likely monomeric. It was extremely sensitive to trypsin. When reconstituted into black lipid bilayer membranes, it formed pores with an average single-channel conductance of 4.6 nanosiemens in 1 M KCl. Zero-current potential measurements showed only a very slight preference, if any, for cations over anions. The data are compatible with pb2 forming a large water-filled transmembrane channel. The functioning during infection of pb2 in cytoplasmic membrane depolarization and phage DNA uptake into the cell is discussed.

Irreversible binding of bacteriophage T5 to its FhuA receptor protein is associated with covalent cross-linking of 3 copies of tail protein pb4.

Irreversible binding of bacteriophage T5 to its FhuA receptor protein is characterized by a high activation energy, typical for reactions where covalent bonds are formed [Zarnitz, M.L. and Weidel, W. (1963) Z. Naturforsch. 18b, 276-280]. Upon binding of radiolabeled T5 phages to FhuA formation of a new protein of 250 kDa was observed. Using electrophoretical and Western blotting techniques this protein was shown to be formed by cross-linking of 3 copies of tail protein pb4, rather than by cross-linking of FhuA and the receptor-binding protein.