[Reduction of decentration after LASIK using a modified eye tracker ring for the MEL-70 excimer laser]. / Reduktion der Dezentrierungsrate nach Lasik durch Weiterentwicklung und Modifikation des Eye-tracker-Rings für MEL-70-Excimer-Laser.

BACKGROUND: The aim of this study was to determine and compare the rate of eccentric laser ablation after LASIK depending on the eye tracker ring used. PATIENTS AND METHODS: All LASIK treatments were carried out using the MEL-70 flying spot excimer laser (Zeiss-Meditec, Jena). The flap was produced using a Corneal Shaper trade mark or Hansatome trade mark Microkeratome (B and L Surgical, Heidelberg). Initially we used an 11 mm eye tracker ring without hinge protector. At the end of February 2001 this ring was replaced by a 10 mm and a 9.5 mm ring with built-in hinge protector. An additional modification was introduced by us: at 1 mm separations little teeth-like spikes were engraved into the eyeward side of the ring, thus stabilising the position of the ring on the globe and allowing free liquid to flow through the spaces between each spike. The built-in calibration system of the corneal topography (TMS 3, Tomey, Erlangen) from patients with a follow-up of one month or longer was used to determine the distance between the centre of the ablation zone from the fixation point. RESULTS: In group I patients (old ring) 42 eyes were treated. In 4 eyes ablation was perfect, in 21 eyes the ablation centre was located 0.1 to 0.49 mm from the fixation point, in 11 eyes 0.51 to 0.99 mm and in 5 eyes 1.1 to 1.49 mm whereas one eye showed a decentred ablation of 1.53 mm. In group II (new ring) 42 eyes were investigated also. In 11 eyes ablation was perfect, in 20 eyes the ablation centre was located 0.1 to 0.49 mm from the fixation point, in 10 eyes 0.5 to 0.99 mm and one eye had an eccentric ablation of 1.28 mm from the fixation point. CONCLUSION: The further development of our eye tracker ring for the MEL-70 laser considerably reduced the rate of decentred ablations. An enhanced grip of the ring onto the globe reduces a slow slide during the laser procedure.

[Measurement of scotopic pupils: comparison of scotic pupil measurements using a colvard pupilometer and the slitlamp green light test]. / Vermessung der skotopischen PupilleEin Vergleich zwischen dem Pupillometer nach Colvard und der Spaltlampenmessung mit dem grünen Licht.

PURPOSE: The aim of the study was to compare the accuracy and the reproducibility of scotopic pupil measurements using two different methods. PATIENTS AND METHODS: We developed a simple test to measure scotopic pupil diameters using a narrow slit of light through a green filter of the Haag-Streit slitlamp. A total of 100 eyes from 50 refractive surgery candidates were prospectively examined by 2 independent investigators using both the Colvard pupilometer (Oasis/USA) and Sekundo's slitlamp green light test. Results were compared using all pairwise multiple comparison procedures (Student-Newman-Keuls method, SigmaStat/Jandel Scientific). The mean age of the patients was 36.3 years with a male:female ratio of 16:34. The colour of the iris was considered blue for 36 individuals and brown for the remainder. RESULTS: Reproducibility: the right eye mean pupil diameter using Sekundo's method was 6.4 mm (+/-0.9) measured by both the first (Slit 1R) and the second (Slit 2R) investigator. The left eye mean pupil diameter was 6.5 mm (+/-1) (Slit 1L) and 6.35 mm (+/-1) (Slit 2L), respectively. The following measurements were obtained with the Colvard pupillometer: (Colv1R)=6.25+/-0.85 mm, (Colv2R)=5.99+/-1 mm, (Colv1L)=6.15+/-0.91 mm, (Colv2L)=6.05+/-0.9 mm. There were no statistically significant differences between the two investigators within the same method of examination. Comparability:The data of both investigators were combined to form 4 groups: (Slit R)=6.4+/-0.96 mm, (Slit L)=6.4+/-1 mm, (Colv R)=6.05+/-0.9 mm, (Colv L)=6.1+/-0.93 mm. There was a significant difference ( p=0.024) between the two methods for both right and left eyes. Particularly for large pupils, the slitlamp green light measurements were significantly higher (up to 0.35 mm) than those with Colvard's device. CONCLUSIONS: The slitlamp green light test provides similar reproducibility for the measurement of the scotopic pupil diameter as the commercially available Colvard pupilometer.

[Therapy of radiation resistant malignant uveal melanoma with endoresection by pars plana vitrectomy in two patients]. / Therapie strahlenresistenter Aderhautmelanome durch Endoresektion im Rahmen einer Pars-plana-Vitrektomie bei zwei Patienten.

BACKGROUND: Since introduction of brachytherapy using (106)ruthenium plaque therapy for treating malignant choroidal melanomas a number of comparative studies have shown that survival rates are equal between patients who underwent radiation or enucleation. When radiation treatment fails, the patient might be determined to choose removal of the tumor by pars plana vitrectomy. PATIENTS AND METHODS: Between 1995 and 1998 a total number of 48 patients underwent (106)ruthenium plaque irradiation. In 9 patients tumor regression was not sufficient. Two of these nine patients decided explicitly against enucleation and choose to have the tumor removed from within the eye employing pars plana vitrectomy. RESULTS: Intraoperatively, the previously radiated melanoma was easily removed with the vitrectomy cutter without significant bleeding. Postoperative recovery was without complications. Silicone oil removal was carried out in one patient after one year. Vision 9 months and 2.5 years after surgery was 0.2 and 0.02, respectively. The cosmetic results with parallel axis of the globes were very good and one patient gained binocular vision. No recurrent tumor growth was seen so far. Histologically, no mitotic figures were found. CONCLUSION: Malignant choroidal melanomas can be safely removed by pars plana vitrectomy with stable intraocular pressure. Previous radiation therapy alters the tumor, inducing fibrous tissue growth and necrosis of tumor cells. These tissue types are not prone to bleed when they are resected by the vitrectomy cutter. Long time studies have to elucidate the difference in survival rates between patients undergoing enucleation of the eye and patients who chose endoresection of the melanoma by pars plana vitrectomy.

Postoperative altered plasma growth hormone levels--a predictor for postoperative complications? A case report.

We compared the onset of predictors for postoperative complications (lactate, total T2 (tT2), total T4 (tT4) and cortisone) retrospectively with the onset of altered growth hormone (GH) concentration in a patient who had had a lethal postoperative outcome and in 13 patients who were without postoperative complications for a period of 24 hours postoperatively. Compared with the values of the patients without postoperative complications, GH values were elevated (68-fold) 1 h after surgery to 103 ng/ml and lactate was increased (12-fold) to 12.7 mmol/l at 6 h postoperatively in the patient with the lethal outcome. The other parameters measured (tT3, tT4 and cortisone) showed no rapid alteration during the first hours postoperatively. This case report suggests that the rapid postoperative onset of raised GH concentration in plasma may be an earlier marker for postoperative complications than the 'established' predictors.

[Self-sealing, sutureless sclerotomy in pars plana vitrectomy]. / Selbstschliessende, nahtlose Sklerotomie zur Pars-plana-Vitrektomie.

BACKGROUND: If sutures are used in the regular closure of sclerotomies a trauma to the bulbus can be inflicted and intraocular bleeding might result. Phases of intraocular hypotony accompany the intraoperative exchange of instruments. Like in no-stitch cataract surgery we employed this type of self-sealing wound closure in pars-plana vitrectomies. A less traumatizing technique yielding a tight and effective closure is an alternative to the conventional approach and results of our experiences are presented. MATERIALS AND METHODS: A modified approach for sutureless sclerotomy closure was applied in 50 eyes (150 sclerotomies). A sclera-covered sclerotomy is performed after preparation of adequate scleral pouches in conventional positions. A minimal conjunctival peritomy allows at the end of surgery an electric cauterization, thus using no suture material in the whole course of vitrectomy. RESULTS: Sutureless vitrectomies can only be installed in the primary operation. However, re-operations on 3 eyes were possible in the same manner using the old sclerotomy sites up to 6 weeks after initial surgery. In 12 eyes the sclerotomy had to be covered with a single suture to obtain adequate wound closure. A repetitive change of instruments during the surgical procedure is possible with this technique and all types of intraocular instruments can be employed. CONCLUSIONS: Self-sealing sclerotomies are a simple and atraumatic approach for wound closure in pars-plana vitrectomies and allow a control of intraocular pressure during surgery.

Postoperative pattern of various hormonal and metabolic variables. A pilot study in patients without complications following cardiac surgery.

The use of single predictors for threatening postoperative complications are widely accepted. However, a typical pattern of multiple parameters could be more helpful than a single predictor. To study this hypothesis, various variables of normal postoperative changes in patients without postoperative complications were investigated. Secondly, this pattern needs to be compared in the future with those findings in patients with postoperative complications. Blood parameters of 13 patients undergoing cardiovascular surgery without postoperative complications for 24 hours were evaluated. Samples were obtained on the afternoon before the operation and 1, 3, 6, 12 and 24 hours after the end of surgery. At one hour postoperation increased levels of the following parameters were noted: growth hormone (p < 0.0001), glucose (p < 0.0001), insulin (p < 0.001), c-peptide (p < 0.001), lactate (p < 0.002), glutamate (p < 0.0001), aspartate (p < 0.001) and total amino acids (p < 0.05), although the concentration of some amino acids decreased. Three hours postoperatively free fatty acids (p < 0.05) were increased. Total-T3 concentrations were reduced postoperatively. Other parameters were not altered. Most of the parameters returned to normal values during the period of observation.

Intercellular signaling by lysophosphatidate.

Lysophosphatidate (LPA) is an intercellular phospholipid messenger with a wide range of biologic effects. The first discovered source of LPA in the human body were activated platelets, but several other sites of LPA generation are now known. The number of cellular interactions is also growing steadily and responses to the compound range wide, from induction of mitogenesis to neurite retraction. LPA acts via a specific G protein-coupled receptor, of which one or more subtypes may exist. Intracellularly, this receptor activates several heterotrimeric G proteins. LPA induces cell proliferation via the small GTP-binding proteins ras, and triggers actin-based cytoskeletal events through rho. This review describes the most relevant recent developments in our understanding of LPA signaling.

Influence of volatile anesthetics on thromboxane A2 signaling.

BACKGROUND: Thromboxane A2 (TXA2) is a member of the prostaglandin family; activation of its receptor induces several important effects, including platelet aggregation and smooth muscle contraction. Because volatile anesthetics interfere with aggregation and contraction, the authors investigated effects of halothane, isoflurane, and sevoflurane on TXA2 signaling in an isolated receptor model. METHODS: mRNA encoding TXA2 receptors was prepared in vitro and expressed in Xenopus oocytes. The effects of halothane, isoflurane, and sevoflurane on Ca2+-activated Cl- currents induced by the TXA2 agonist U-46619 and on those induced by intracellular injection of inositol 1-4-5 trisphosphate or guanosine 5'-O-(2-thiodiphosphate) were measured using the voltage-clamp technique. RESULTS: Expressed TXA2 receptors were functional (half maximal effect concentration [EC50], 3.2 x 10(-7) +/- 1.1 x 10(-7) M; Hill coefficient (h), 0.8 +/- 0.2). Halothane and isoflurane inhibition of TXA2 signaling was reversible and concentration dependent (halothane half maximal inhibitory concentration [IC50], 0.46 +/- 0.04 mM; h, 1.6 +/- 0.21; isoflurane IC50, 0.69 +/- 0.12 mM; h, 1.3 +/- 0.27). 0.56 mM halothane (1%) right-shifted the U-46619 concentration-response relationship by two orders of magnitude (EC50, 1 x 10[-5] M). That h and maximal effect (Emax) were unchanged indicates that halothane acts in a competitive manner. In contrast, isoflurane acted noncompetitively, decreasing Emax by 30% (h and EC50 were unchanged). Both halothane and isoflurane had no effect on intracellular signaling pathways. Sevoflurane (0-1.3 mM) did not affect TXA2 signaling. CONCLUSIONS: Both halothane and isoflurane inhibit TXA2 signaling at the membrane receptor, but by different mechanisms. This suggests that the effects of these anesthetics on TXA2 signaling are evoked at different locations of the receptor protein: halothane probably acts at the ligand binding site and isoflurane at an allosteric site.

Volatile anaesthetics have differential effects on recombinant m1 and m3 muscarinic acetylcholine receptor function.

Muscarinic acetylcholine signalling plays major roles in regulation of consciousness, cognitive functioning, pain perception and circulatory homeostasis. Halothane has been shown to inhibit m1 muscarinic signalling. However, no comparative data are available for desflurane, sevoflurane or isoflurane, nor have the anaesthetic effects on the m3 subtype (which is also prominent in the brain) been studied. Therefore, we have investigated the effects of these compounds on isolated m1 and m3 muscarinic receptor function. Defolliculated Xenopus oocytes expressing recombinant m1 or m3 muscarinic or (for comparison) AT1A angiotensin II receptors were voltage clamped, and Ca(2+)-activated Cl- currents (ICl(Ca)) induced by acetyl-beta-methylcholine (Mch) or angiotensin II were measured in the presence of clinically relevant concentrations of halothane, sevoflurane, desflurane or isoflurane. To determine the site of action of the volatile anaesthetics we compared anaesthetic effects on m1, m3 and AT1A receptor function and studied the effects of volatile anaesthetics on signalling induced by intracellular injection of the second messenger IP3. Desflurane had a biphasic effect on m1 signalling, enhancing at a concentration of 0.46 mmol litre-1 but depressing at 0.92 mmol litre-1. A similar, although not significant, trend was observed with m3 signalling. Isoflurane had no effect on m1 signalling, but profoundly inhibited m3 signalling. Sevoflurane depressed the function of m1 and m3 signalling in a dose-dependent manner. Halothane, similar to its known effect on m1 signalling, dose-dependently depressed m3 function. ICl(Ca) induced by intracellular injections of IP3 were unaffected by all four anaesthetics. Similarly, none of the anaesthetics tested affected AT1A signalling. Absence of interference with AT1A signalling and intracellular pathways suggest that the effects of anaesthetics on muscarinic signalling most likely result from interactions with the m1 or m3 receptor molecule. Multiple interaction sites with different affinities may explain the biphasic response to desflurane. Anaesthetic-specific effects on closely related receptor subtypes suggest defined sites of action for volatile anaesthetics on the receptor protein.

Influence of anesthetics on endogenous and recombinantly expressed G protein-coupled receptors in the Xenopus oocyte.

1. The oocyte of the African clawed toad (Xenopus laevis) offers a reliable, sensitive and disease resistant system to investigate recombinantly and endogenously expressed Ca2+ signaling G protein-coupled receptors and their intracellular signaling pathways. 2. To study receptor induced Ca2+ release, two-electrode voltage clamping can quantify a Ca2+-activated transmembrane Cl- current. Intracellular steps of the signaling pathway can be inhibited by injections of EDTA or heparin into the oocyte. Components of the intracellular pathway can be activated directly by GTPgammaS or IP3 injection. 3. We have investigated the effects of volatile, local and i.v. anesthetics on the signaling properties of the endogenous lysophosphatidate receptor and on mammalian receptors expressed recombinantly by intracellular injection of the encoding mRNA or cDNA. A number of receptors are sensitive to these anesthetics. Anesthetics interact with muscarinic, thromboxane A2 and lysophosphatidate signaling. 4. Investigations of the intracellular pathways revealed that the receptor or the receptor-G protein coupling is affected primarily and that mechanisms further downstream are not influenced by the various types of anesthetics.

Synergistic inhibition of muscarinic signaling by ketamine stereoisomers and the preservative benzethonium chloride.

BACKGROUND: Ketamine (Ketalar; Parke-Davis, Morris Plains, NJ) has been shown to inhibit muscarinic signaling with a median inhibitory concentration (IC50) of 5.7 microM. Whereas Ketalar is a racemic mixture, recent interest has focused on clinical use of the S(+) ketamine isomer, which is three times as potent an analgesic as the R(-) isomer yet seems to be associated with fewer psychoactive side effects. Therefore, the authors studied the effects of S(+) and R(-) ketamine and the preservative benzethonium chloride on muscarinic signaling. METHODS: Rat ml muscarinic acetylcholine receptors were expressed recombinantly in Xenopus laevis oocytes. Ca2(+)-activated Cl- currents in response to 10(-7) M acetyl-beta-methylcholine were determined by two-electrode voltage clamping in the presence of various concentrations of ketamine and benzethonium. Concentration-inhibition curves were constructed and used for algebraic and isobolographic analysis. RESULTS: The IC50. was 125 +/- 33 microM for S(+) ketamine, and 91 +/- 19 microM for R(-) ketamine. This difference was not statistically significant, indicating that muscarinic inhibition by ketamine is not stereoselective. The R(-)/S(+) mixture had an IC50 of 48 +/- 1 microM, and thus the stereoisomers interact synergistically. When appropriate concentrations of benzethonium were added, an IC50 of 15 +/- 2 microM resulted. CONCLUSIONS: The muscarinic inhibitory action of ketamine isomers is not stereoselective. Because S(+) ketamine is a significantly more potent analgesic, it should have less muscarinic inhibitory action than R(-) ketamine when used in clinically equivalent doses. A significant fraction of the muscarinic inhibitory action of Ketalar is due to the preservative benzethonium. If reconstituted with a different preservative, Ketalar might be a less potent muscarinic antagonist.

Inhibition of lysophosphatidate signaling by lidocaine and bupivacaine.

BACKGROUND: Lidocaine and bupivacaine impair wound healing, but the mechanism of this side effect has not been determined. The phospholipid messenger lysophosphatidate is released from activated platelets and induces fibroblast and smooth muscle proliferation. Because it may play a role in wound healing, the authors studied the effects of local anesthetics on lysophosphatidate signaling in Xenopus oocytes. METHODS: Defolliculated Xenopus oocytes expressing endogenous G protein-coupled lysophosphatidate receptors were voltage clamped and studied in the presence or absence of lidocaine or bupivacaine. Lysophosphatidate-induced Ca(2+)-activated Cl- currents (IC1(Ca)) were measured. To determine the site of action of the local anesthetics on the signaling pathway, the authors studied 1) the effects of local anesthetics on signaling induced by intracellular injection of the second messenger inositoltrisphosphate, and 2) the effects of local anesthetics on functioning of recombinantly expressed angiotensin II receptor signaling through the same pathways as the lysophosphatidate receptor. RESULTS: Lysophosphatidate signaling was inhibited in the presence of local anesthetics. The half maximal inhibitory concentration (IC50S) for lidocaine and bupivacaine were 29.6 mM and 4.7 mM, respectively. Neither responses induced by inositoltrisphosphate injection nor angiotensin signaling were influenced by local anesthetics. CONCLUSIONS: Lysophosphatidate signaling is inhibited by the extracellular application of lidocaine or bupivacaine. In contrast, inositoltrisphosphate or angiotensin signaling was not affected by local anesthetics. Therefore local anesthetics have a specific, extracellular effect on lysophosphatidate receptor functioning. As the local anesthetic concentrations used were similar to those observed after injection around surgical wounds, LP inhibition may play a role in the observed detrimental effects of local anesthetics on wound healing.

Detection of numerical karyotype changes in the giant cells of Hodgkin's lymphomas by a combination of FISH and immunohistochemistry applied to paraffin sections.

Conventional karyotyping of Hodgkin's disease (HD) has until now yielded only limited insight into karyotypic characteristics of this disease. For this reason, fluorescence in situ hybridization (FISH) using alpha-satellite chromosome-specific probes was applied to paraffin sections of HD tumors in order to verify numerical aberrations suggested to be specific for HD in the literature. The FISH technique was combined with immunohistochemical detection of the CD30 antigen to allow easier identification of the Reed-Sternberg and Hodgkin (RS&H) cells. The number of specific FISH signals per nucleus was determined both in CD30-positive RS&H cells as well as in non-malignant "bystander" cells in order to assess differences in the signal distribution. Contrasted with normal lymphoid cells, the tumor cells in HD were found to be polysomic for at least one of the chromosomes analyzed (1,2,4, and 8). The technique described is a reliable method for confirmation of results obtained from conventional cytogenetics, which is especially suited for archival material or samples not containing dividing cells.

[Neuromuscular blockade after atracurium and alcuronium with propofol and thiopental]. / Neuromuskuläre Blockade nach Atracurium und Alcuronium unter dem Einfluss von Propofol und Thiopental.

UNLABELLED: Does propofol or thiopentone enhance the effect of nondepolarizing muscle relaxants? We evaluated the effects of propofol and thiopentone on the pharmacodynamics of atracurium and alcuronium in 43 surgical patients (ASA I and II) under general anaesthesia. METHODS: The patients were randomized into five groups, A-E. Anaesthesia was induced in all patients with fentanyl 4 micrograms/kg i.v. Patients in groups A and C patients received thiopentone 7 mg/kg i.v., and relaxation was achieved with alcuronium 0.25 mg/kg (group A) and atracurium 0.5 mg/kg (group C). Electromyography (train of four, TOF) was used to determine the time of onset of relaxation (AZ) and the maximum degree of blockade (T%). The recovery times to 25%, 50% and 75% of baseline muscle strength were recorded. Additionally, the TOF ratio T4:T1 was calculated, indicating the probable end of relaxation at a ratio of 0.7. At the beginning of the recovery phase (T1 = 15%) propofol 1% 3 mg/kg was given, and the effect on the TOF was measured. Patients in groups B and D patients received total intravenous anaesthesia (TIVA) with propofol 1% 6-12 mg/kg per hour continuously after induction with 3 mg/kg. The action profile of alcuronium 0.25 mg/kg (group B) and atracurium 0.5 mg/kg (group D) were recorded. Group E patients received thiopentone (10 mg/kg per hour) under the use of atracurium 0.5 mg/kg. Ventilation was performed with 30%/70% oxygen and N2O. The results were analyzed for significance using the Mann-Whitney U-test (P = 0.019). RESULTS: A slight difference in AZ was noted for alcuronium under the use of TIVA between propofol and thiopentone: 13 min and 5 min, respectively. Otherwise, the pharmacodynamics (T% and recovery of neuromuscular function) of the two relaxants exhibited no major differences related to thiopentone, propofol or their combination. The TOF was not influenced under additional propofol application. Noteworthy were the wide distribution of the time course of action (up to 3 h) and the magnitude of T% depression under alcuronium. CONCLUSION: Propofol and thiopentone have no potentiating influence on the time course of action and the magnitude of relaxation with alcuronium and atracurium. Pharmacodynamics of nondepolarizing muscle relaxants do not seem to be influenced by these two hypnotics.

Intrahepatic bile duct loss in biliary atresia despite portoenterostomy: a consequence of ongoing obstruction?

The histological and immunohistochemical characteristics of the liver in 44 children (28 boys, 16 girls) with extrahepatic biliary atresia at different stages of the clinical course were studied. Thirty-four wedge liver biopsy specimens taken during Kasai operations (25 specimens) and relaparotomy (9 specimens) and 20 hepatectomy explants taken at the time of transplantation were examined. Routine histological stains and monoclonal antibodies against different molecular weight cytokeratins and HLA-DR were used. The histopathological changes and the pattern of cytokeratin expression observed during the course of the disease were suggestive of persistent or recurrent extrahepatic biliary obstruction that occurred despite the Kasai operation and eventually led to cirrhosis and liver failure. Quantitative studies showed a progressive loss of intrahepatic bile ducts over the time course of the disease. This destruction of bile ducts had a geographic anatomical distribution in hepatectomy specimens, and in two livers it occurred predominantly in only one lobe. This geographic distribution of the vanishing bile ducts probably indicates an unpredictable and uneven obliteration of bile ducts in the porta hepatis during portoenterostomy wound healing and scarring.