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1.
Indian J Med Microbiol ; 26(1): 13-20, 2008.
Article in English | MEDLINE | ID: mdl-18227591

ABSTRACT

PURPOSE: Molecular genotyping relies on the identification of specific microbial DNA sequences. Accurate genotyping not only requires discrimination between low- and high-risk pathogens for effective diagnosis or disease management but also requires the identity of the specific strain or type of the microbe involved in pathogenesis. The majority of these assays require DNA amplification followed by genome identification either through sequencing or hybridization to specific oligonucleotide probes. We evaluated the use of a DNA microchip assay as a simple and easy-to-use procedure for genotyping. METHODS: Various methodological parameters were optimized for single-base mismatch discrimination on a DNA microarray. The fabrication procedures involved substrate chemistry for immobilization. The effect of various buffers and features associated with oligonucleotide sequences were standardized. The assay was evaluated on a low-density genotyping chip containing the sequences of various (Human Papilloma Virus) HPV subtypes. RESULTS: The specific subtype was identified with high specificity by hybridization in miniaturized condition. CONCLUSIONS: The DNA microchip provides a rapid and cost-effective genotyping procedure for microbial organisms and can be implemented easily in any laboratory.


Subject(s)
Molecular Diagnostic Techniques/methods , Oligonucleotide Array Sequence Analysis/methods , Papillomaviridae/classification , Papillomaviridae/isolation & purification , Papillomavirus Infections/diagnosis , Papillomavirus Infections/virology , Genotype , Humans , Molecular Diagnostic Techniques/standards , Oligonucleotide Array Sequence Analysis/standards , Sensitivity and Specificity
2.
J Neurosurg Anesthesiol ; 13(3): 222-6, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11426096

ABSTRACT

Previous studies have shown that laparoscopic interventions are associated with increases in intracranial pressure. However, the consequences on cerebral blood flow (CBF) are unknown. This study investigates the effects of carbon dioxide (CO2) pneumoperitoneum on CBF in pigs. Ten pigs (weight, 20-26 kg) were anesthetized with 1.4% isoflurane and fentanyl (1 microg/kg per minute). Mechanical ventilation (fraction of inspired oxygen = 0.4) was set to maintain normocapnia (end-tidal CO2 tension = 35 mm Hg). Arterial and central venous catheters were placed for measurement of mean arterial blood pressure and central venous pressure. Bilateral internal carotid artery blood flow was measured using two transient time flow probes placed around both carotid arteries (with ligated external carotid arteries). Cortical and subcortical cerebral blood flow was measured using laser Doppler flowmetry. Sagittal sinus pressure was measured via a superior sagittal sinus catheter. After baseline measurements, the peritoneal cavity of the animals was insufflated with CO2 to achieve an intraabdominal pressure of 12-mm Hg. After 10 minutes of stable CO2, pneumoperitoneum measurements were repeated. Increases in central venous pressure (6.3 +/- 2.1 to 11.1 +/- 3.0 mm Hg) and sagittal sinus pressure (8.0 +/- 2.8 to 11.9 +/- 3.0 mm Hg) were noted during CO2 pneumoperitoneum (P < .05). Bilateral internal carotid artery blood flow (46.0 +/- 7.4 vs 47.7 +/- 7.1 mL/100g per minute), cortical CBF (263 +/- 115 vs 259 +/- 158 tissue perfusion units), and subcortical CBF (131 +/- 145 vs 133 +/- 149 tissue perfusion units) did not change during CO2 pneumoperitoneum. The current data show that CO2 pneumoperitoneum increases sagittal sinus pressure without changing CBF. Increases in sagittal sinus pressure are likely related to decreases in cerebral venous drainage caused by increases in intraabdominal pressure.


Subject(s)
Carbon Dioxide/blood , Cerebrovascular Circulation/physiology , Hemodynamics/physiology , Pneumoperitoneum/physiopathology , Abdomen , Acid-Base Equilibrium/physiology , Animals , Blood Pressure/physiology , Brain/blood supply , Carbon Dioxide/administration & dosage , Central Venous Pressure/physiology , Heart Rate/physiology , Intracranial Pressure/physiology , Pressure , Regional Blood Flow , Swine , Time Factors
3.
Nat Cell Biol ; 3(4): 392-9, 2001 Apr.
Article in English | MEDLINE | ID: mdl-11283613

ABSTRACT

Deregulated expression of c-myc can induce cell proliferation in established cell lines and in primary mouse embryonic fibroblasts (MEFs), through a combination of both transcriptional activation and repression by Myc. Here we show that a Myc-associated transcription factor, Miz-1, arrests cells in G1 phase and inhibits cyclin D-associated kinase activity. Miz-1 upregulates expression of the cyclin-dependent kinases (CDK) inhibitor p15INK4b by binding to the initiator element of the p15INK4b promoter. Myc and Max form a complex with Miz-1 at the p15 initiator and inhibit transcriptional activation by Miz-1. Expression of Myc in primary cells inhibits the accumulation of p15INK4b that is associated with cellular senescence; conversely, deletion of c-myc in an established cell line activates p15INK4b expression. Alleles of c-myc that are unable to bind to Miz-1 fail to inhibit accumulation of p15INK4b messenger RNA in primary cells and are, as a consequence, deficient in immortalization.


Subject(s)
Carrier Proteins/biosynthesis , Cell Cycle Proteins , Cyclin-Dependent Kinase Inhibitor p16 , DNA-Binding Proteins/metabolism , Proto-Oncogene Proteins c-myc/metabolism , Transcription Factors/metabolism , Tumor Suppressor Proteins , Zinc Fingers , 3T3 Cells , Animals , Cyclin-Dependent Kinase Inhibitor p15 , DNA-Binding Proteins/genetics , Gene Expression Regulation , HeLa Cells , Humans , Kruppel-Like Transcription Factors , Mice , Nuclear Proteins/metabolism , Promoter Regions, Genetic , Rats , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Trans-Activators/metabolism , Transcription Factors/genetics
4.
Anesthesiology ; 93(3): 793-804, 2000 Sep.
Article in English | MEDLINE | ID: mdl-10969313

ABSTRACT

BACKGROUND: The aim of this study was to histologically investigate brain damage after prolonged periods of bacteremia in pigs. METHODS: Twenty-one pathogen-free Göttingen minipigs were anesthetized and instrumented with a femoral arterial, a pulmonary arterial, and through midline abdominal incision with a portal venous catheter. After craniotomy the superior sagittal sinus was cannulated. A lumbosacral spinal catheter was inserted for sampling of cerebrospinal fluid. Twelve hours after instrumentation, the animals were randomized in two groups: septic and control animals. The septic group received an infusion of 107 colony-forming units per kilogram of living Escherichia coli over 0.5 h through portal venous catheter each day. The control group received saline. Postoperative intensive care treatment included 4 days of controlled mechanical ventilation, sedation, and intravenous nutrition. The brains then were removed, fixed, and processed for histology. Each pathologic alteration found in the samples was assessed and given a severity code (0-3). RESULTS: Sham-operated animals showed no alterations caused by the instrumentation and the intensive care treatment. The septic group showed typical clinical signs of sepsis. Vasopressor support and mechanical ventilation prevented systemic hypotension and hypoxemia. High serum and cerebrospinal fluid levels of interleukin-6 and tumor necrosis factor-alpha were detected. The septic group showed severe histologic abnormalities of the brain including perivascular edema, spongiform degeneration, hyperemia, and purpura. Damage of neurons was seen including eosinophilic cytoplasm, shrunken nuclei, and disintegration of the nuclear membrane. CONCLUSIONS: Abdominal sepsis induced severe brain damage that was not related to systemic hypoxia or ischemia. High cerebrospinal fluid levels of tumor necrosis factor-alpha and interleukin-6 were related to an inflammatory process in the brain resulting in cerebral edema and death of neurons.


Subject(s)
Bacteremia/pathology , Brain/pathology , Animals , Bacteremia/metabolism , Body Temperature , Brain/metabolism , Brain Edema/etiology , Chronic Disease , Female , Interleukin-6/biosynthesis , Portal Vein , Swine , Tumor Necrosis Factor-alpha/biosynthesis
5.
Br J Anaesth ; 84(2): 221-5, 2000 Feb.
Article in English | MEDLINE | ID: mdl-10743456

ABSTRACT

We have investigated the effects of xenon on regional cerebral blood flow (rCBF) and autoregulation in pigs sedated with propofol 4 mg kg-1 h-1. Balloon-tipped catheters were placed into the descending aorta and inferior vena cava of 15 Göttingen Minipigs for manipulation of arterial pressure and blood sampling. rCBF was measured using the sagittal sinus outflow technique. Xenon was adjusted randomly to end-tidal fractions (FE'Xe) of 0, 0.30, 0.50 and 0.70. After baseline measurements of heart rate (HR), mean arterial pressure (MAP), rCBF, sagittal sinus pressure (SSP) and calculation of regional cerebrovascular resistance (rCVR) at each respective FE'Xe, autoregulation was tested in the MAP range 60-120 mm Hg. Increasing FE'Xe had no effect on HR, MAP, rCBF or SSP. rCVR increased with increases in MAP, regardless of FE'Xe. Autoregulation was not impaired. We conclude that xenon inhalation had no effect on rCBF and autoregulation in our model, which could suggest that xenon is an adequate adjunct for neurosurgical anaesthesia.


Subject(s)
Anesthetics, Inhalation/pharmacology , Cerebrovascular Circulation/drug effects , Homeostasis/drug effects , Xenon/pharmacology , Anesthetics, Intravenous , Animals , Female , Hemodynamics/drug effects , Propofol , Swine , Swine, Miniature , Vascular Resistance/drug effects
6.
Biotechniques ; 27(2): 368-75, 1999 Aug.
Article in English | MEDLINE | ID: mdl-10457846

ABSTRACT

The c-Myc protein, the product of the c-myc protooncogene, is a nuclear phosphoprotein with DNA-binding properties when heterodimerized with the Max protein. It contains an amino-terminal transcriptional activation domain and a carboxy-terminal basic helix-loop-helix leucine zipper (bHLHzip) domain that directs heterodimerization and promotes DNA binding. Here, we describe the isolation of the bHLHzip domain of human c-Myc with a technique for efficient single-step purification. Using a C-terminal Strep-tag II affinity peptide and a novel Streptactin-Sepharose matrix, elution is performed under mild conditions by competition with the biotin analog desthiobiotin. No significant influence of the affinity tag on the activity of the bHLHzip domain was observed when the fusion protein was subjected to glutathione S-transferase (GST) pull-down assays for investigating its in vitro-binding properties with GST-Max. The use of the C-terminal Strep-tag II was shown to be more suitable for obtaining pure product fractions than use of the N-terminal GST affinity tag.


Subject(s)
Chromatography, Affinity/methods , Helix-Loop-Helix Motifs , Proto-Oncogene Proteins c-myc/isolation & purification , Transcription Factors , Bacterial Outer Membrane Proteins/chemistry , Basic Helix-Loop-Helix Leucine Zipper Transcription Factors , Basic-Leucine Zipper Transcription Factors , DNA-Binding Proteins/chemistry , Glutathione Transferase/isolation & purification , Humans , Leucine Zippers , Protein Conformation , Proto-Oncogene Proteins c-myc/chemistry , Recombinant Fusion Proteins/chemistry , Recombinant Fusion Proteins/isolation & purification , Streptavidin/chemistry
7.
Article in German | MEDLINE | ID: mdl-10234406

ABSTRACT

Impaired mental function, from clouding of consciousness to deep coma is often seen in patients with systemic inflammation. Diagnosis of this syndrome which is called "septic encephalopathy" is dependent on exclusion of other causes. The underlying mechanisms have only been defined in parts. The appearance of cerebral symptoms during an infection increases mortality. Primary symptoms of septic encephalopathy appear early, before other septic organ manifestations become apparent. The most sensible parameter for diagnosis of septic encephalopathy in comatose patients or under sedation is the EEG. It shows general alterations which increase parallelly to the severity of septic encephalopathy. Septic encephalopathy has to be considered a multifactorial event. In an early stage of the development of septic encephalopathy, bacteremia induces overproduction of cytokines and other mediators. This causes metabolic dysregulation with effects on the cerebral protein-, glucose and neurotransmitter metabolism. In addition, cytokines damage the blood-brain-barrier and exert direct cytotoxic effects. This results in histologic detectable neuronal damage. Further effects of the cytokine expression are perivascular edema and hemorrhage. The loss of metabolic regulation of the brain perfusion and local cerebral ischemia additionally contribute to the etiology of septic encephalopathy. A specific therapy is not yet known.


Subject(s)
Brain Diseases/microbiology , Sepsis/microbiology , Brain Diseases/epidemiology , Brain Diseases/psychology , Humans , Sepsis/psychology
8.
Proc Natl Acad Sci U S A ; 96(5): 1941-6, 1999 Mar 02.
Article in English | MEDLINE | ID: mdl-10051574

ABSTRACT

The Mg-chelation is found to be a prerequisite to direct protoporphyrin IX into the chlorophyll (Chl)-synthesizing branch of the tetrapyrrol pathway. The ATP-dependent insertion of magnesium into protoporphyrin IX is catalyzed by the enzyme Mg-chelatase, which consists of three protein subunits (CHL D, CHL I, and CHL H). We have chosen the Mg-chelatase from tobacco to obtain more information about the mode of molecular action of this complex enzyme by elucidating the interactions in vitro and in vivo between the central subunit CHL D and subunits CHL I and CHL H. We dissected CHL D in defined peptide fragments and assayed for the essential part of CHL D for protein-protein interaction and enzyme activity. Surprisingly, only a small part of CHL D, i.e., 110 aa, was required for interaction with the partner subunits and maintenance of the enzyme activity. In addition, it could be demonstrated that CHL D is capable of forming homodimers. Moreover, it interacted with both CHL I and CHL H. Our data led to the outline of a two-step model based on the cooperation of the subunits for the chelation process.


Subject(s)
Lyases/chemistry , Lyases/metabolism , Nicotiana/enzymology , Plants, Toxic , Protoporphyrins/metabolism , Chlorophyll/biosynthesis , Cloning, Molecular , Escherichia coli , Kinetics , Macromolecular Substances , Models, Chemical , Models, Molecular , Protein Conformation , Recombinant Fusion Proteins/biosynthesis , Recombinant Proteins/chemistry , Recombinant Proteins/metabolism , Restriction Mapping
9.
Anesthesiology ; 89(1): 19-23, 1998 Jul.
Article in English | MEDLINE | ID: mdl-9667289

ABSTRACT

UNLABELLED: BACKGROUND. The rewarming period of hypothermic cardiopulmonary bypass (CPB) is associated with reduced jugular bulb venous oxygen saturation (SjO2). This study investigates the effects of normocapnia vs. hypercapnia on changes in SjO2 during rewarming from hypothermic CPB for coronary artery bypass graft in patients classified as American Society of Anesthesiologists physical status 111. METHODS: Anesthesia was induced and maintained with fentanyl, midazolam, and continuous infusion of etomidate. Hypothermic CPB (27 degrees C) was managed according to alpha-stat conditions. The SjO2 percentage was measured using a fiberoptic catheter placed in the right jugular bulb via the right internal jugular vein. Data were recorded before and during the rewarming period. Patients were assigned to a normocapnic (PaCO2: 36-40 mmHg, n = 10) or hypercapnic (PaCO2: 45-50 mmHg, n = 10) PaCO2 regimen during rewarming. RESULTS: The maximum reduction of SjO2 occurred during rewarming with the jugular bulb temperature at 35-36 degrees C. In contrast, SjO2 did not change during rewarming from hypothermia in hypercapnic patients. CONCLUSIONS: These results show that mild hypercapnia prevents the desaturation of SjO2 seen with the normocapnic group during the rewarming period from hypothermic CPB. These data suggest that mild hypercapnia during rewarming from CPB is associated with a better balance between cerebral oxygen supply and demand.


Subject(s)
Cardiopulmonary Bypass , Hemoglobins/metabolism , Hypercapnia , Hypothermia, Induced/adverse effects , Intraoperative Complications/prevention & control , Jugular Veins/physiopathology , Aged , Female , Humans , Jugular Veins/metabolism , Male , Middle Aged
10.
J Cardiothorac Vasc Anesth ; 11(6): 689-93, 1997 Oct.
Article in English | MEDLINE | ID: mdl-9327306

ABSTRACT

OBJECTIVE: This study investigates the effects of rapid versus graded rewarming on decreases in jugular bulb oxygen saturation (SjO2) during cardiopulmonary bypass (CPB) in a prospective nonrandomized and nonblinded design. SETTING AND PARTICIPANTS: At the Department of Anesthesiology (University Hospital Eppendorf, Germany), 28 patients (ASA III) undergoing coronary artery bypass graft were investigated. INTERVENTION: CPB was managed according to alpha-stat conditions during moderate hypothermia (27 degrees C). In group 1 (n = 17), rewarming was performed by increasing the perfusate temperature to 36 degrees C within 7 minutes, in group 2 (n = 11) within 15 minutes. MEASUREMENTS AND MAIN RESULTS: SjO2 was measured by a fiberoptic catheter placed in the right jugular bulb. Data were recorded before and 40 minutes after the start of rewarming every 5 minutes. During rewarming of CPB, SjO2 was decreased to 43 +/- 7% in group 1 and to 44 +/- 4% in group 2. In groups 1 and 2, the maximum reduction of SjO2 occurred 17 minutes and 30 minutes after start of rewarming, respectively. The delayed reduction of SjO2 in group 2 correlated strongly with the prolonged increase in jugular bulb temperature. CONCLUSION: The current data show that slow rewarming does not attenuate reductions of SjO2. This suggests that the reduction of SjO2 during rewarming of CPB is not a function of the rewarming speed but is strongly correlated with the increase in jugular bulb temperature, with a maximum effect just before reaching normothermia of the brain.


Subject(s)
Cardiopulmonary Bypass , Hot Temperature , Hypothermia, Induced , Oxygen/blood , Adult , Aged , Body Temperature , Female , Humans , Jugular Veins , Male , Middle Aged , Prospective Studies
11.
EMBO J ; 16(18): 5672-86, 1997 Sep 15.
Article in English | MEDLINE | ID: mdl-9312026

ABSTRACT

The c-Myc protein activates transcription as part of a heteromeric complex with Max. However, Myc-transformed cells are characterized by loss of expression of several genes, suggesting that Myc may also repress gene expression. Two-hybrid cloning identifies a novel POZ domain Zn finger protein (Miz-1; Myc-interacting Zn finger protein-1) that specifically interacts with Myc, but not with Max or USF. Miz-1 binds to start sites of the adenovirus major late and cyclin D1 promoters and activates transcription from both promoters. Miz-1 has a potent growth arrest function. Binding of Myc to Miz-1 requires the helix-loop-helix domain of Myc and a short amphipathic helix located in the carboxy-terminus of Miz-1. Expression of Myc inhibits transactivation, overcomes Miz-1-induced growth arrest and renders Miz-1 insoluble in vivo. These processes depend on Myc and Miz-1 association and on the integrity of the POZ domain of Miz-1, suggesting that Myc binding activates a latent inhibitory function of this domain. Fusion of a nuclear localization signal induces efficient nuclear transport of Miz-1 and impairs the ability of Myc to overcome transcriptional activation and growth arrest by Miz-1. Our data suggest a model for how gene repression by Myc may occur in vivo.


Subject(s)
DNA-Binding Proteins/metabolism , Gene Expression Regulation , Proto-Oncogene Proteins c-myc/metabolism , 3T3 Cells , Adenoviridae/genetics , Amino Acid Sequence , Animals , Base Sequence , Cyclin D1/biosynthesis , Cyclin D1/genetics , DNA-Binding Proteins/biosynthesis , DNA-Binding Proteins/chemistry , DNA-Binding Proteins/genetics , Gene Library , Genes, Reporter , HeLa Cells , Helix-Loop-Helix Motifs , Humans , Kruppel-Like Transcription Factors , Mice , Molecular Sequence Data , Polymerase Chain Reaction , Promoter Regions, Genetic , Protein Biosynthesis , Recombinant Fusion Proteins/biosynthesis , Recombinant Fusion Proteins/metabolism , Transcription Factors/biosynthesis , Transcription Factors/genetics , Transcription, Genetic , Transfection , Viral Proteins , Zinc Fingers
12.
Anesthesiology ; 87(1): 58-62, 1997 Jul.
Article in English | MEDLINE | ID: mdl-9232134

ABSTRACT

BACKGROUND: Transmyocardial laser revascularization may vaporize fluid in the left heart, allowing bubbles to form. This study aimed to determine whether the laser pulse resulted in cerebral emboli and to examine changes in middle cerebral artery flow velocity and jugular bulb oxygen saturation (SjO2) during transmyocardial laser revascularization. METHODS: Twelve patients (American Society of Anesthesiologists physical status III) were studied after the authors received institutional review board approval and the patients' informed consent. Monitored variables included mean arterial blood pressure (measured in millimeters of mercury), heart rate (measured as beats/min), and partial pressure of carbon dioxide (measured in millimeters of mercury). A 5-MHz transesophageal-sonography system was used to record intraventricular events after laser injection. Mean blood flow velocity (Vmean; measured in centimeters per second) was monitored in the middle cerebral artery using transcranial Doppler sonography, and SjO2 (expressed as a percentage) was measured using a fiberoptic thermodilution catheter placed in the right jugular bulb. Data were recorded before, during, and for 4 min after laser injection. RESULTS: After laser injection, intraventricular echogenic contrast was seen in transesophageal-sonography, and 2-4 s later high-intensity signals (microemboli) appeared in the transcranial Doppler sonography spectra. As long as mean arterial pressure remained stable during the observation period, Vmean and SjO2 did not change. CONCLUSIONS: These data show that microemboli can be detected after laser injection in the middle cerebral artery, although they do not effect Vmean and SjO2. The results suggest that these microemboli do not induce a global oxygen imbalance.


Subject(s)
Coronary Disease/surgery , Intracranial Embolism and Thrombosis/etiology , Laser Therapy/adverse effects , Myocardial Revascularization/adverse effects , Aged , Cerebrovascular Circulation , Female , Humans , Male , Middle Aged
13.
J Neurosurg Anesthesiol ; 9(3): 223-7, 1997 Jul.
Article in English | MEDLINE | ID: mdl-9239583

ABSTRACT

Our study investigated the effects of moderate doses of fentanyl and sufentanil versus high-dose sufentanil on cerebral hemodynamics by using transcranial Doppler ultrasonography (TCD). Thirty American Society of Anesthesiologists (ASA) II and III patients scheduled for elective coronary artery bypass graft (CABG) were studied after Institutional Review Board (IRB) approval and informed consent. The evening before surgery, all patients received oral flurazepam (1 mg/kg), Atropine (0.4 mg/70 kg s.c.) and a combination of droperidol (70 micrograms/kg s.c.) plus fentanyl (1.5 micrograms/kg s.c.) were given as preanesthetic medication 1 h before induction of anesthesia. Anesthesia was induced with either 25 micrograms/kg fentanyl i.v. (group 1, n = 10), 3 micrograms/kg sufentanil i.v. (group 2, n = 10) or 6 micrograms/kg sufentanil i.v. (group 3, n = 10). All patients received 100 micrograms/kg pancuronium i.v. With the induction of respiratory depression, assisted ventilation was performed followed by controlled ventilation to maintain normoxia and normocapnia (FiO2, 1.0). Cerebral blood flow velocity (CBFV, cm/s) was measured continuously in the middle cerebral artery by using a bidirectional 2-MHz TCD system. Monitoring included heart rate (HR, beats/min), direct mean arterial blood pressure (MAP, mm Hg), and PaCO2. Physiologic variables including arterial blood gases were measured at baseline, 5 min, and 10 min after infusion of fentanyl or sufentanil. In all patients, HR, MAP, end-tidal carbon dioxide tension (PetCO2), and PaCO2 were constant over time and did not differ between groups. CBFV did not change with moderate doses of fentanyl (group 1) or sufentanil (group 2). In contrast, infusion of high-dose sufentanil (group 3) was associated with 27 to 30% decreases in CBFV (p < 0.05). Our results suggest that sufentanil decreases CBFV in a dose-related fashion with a threshold effect. Increases in CBFV and CBF seen in previous studies may be related to an increasing PaCO2 when maintenance of normocarbia is based on only real-time capnography with a constant PetCo2 rather than additional arterial blood gas monitoring.


Subject(s)
Analgesics, Opioid/pharmacology , Cerebrovascular Circulation/drug effects , Fentanyl/pharmacology , Sufentanil/pharmacology , Carbon Dioxide/blood , Dose-Response Relationship, Drug , Hemodynamics/drug effects , Humans , Ultrasonography, Doppler, Transcranial
14.
J Neurosurg Anesthesiol ; 9(2): 128-33, 1997 Apr.
Article in English | MEDLINE | ID: mdl-9100181

ABSTRACT

This study investigates changes of jugular bulb oxygen saturation (SjO2) measured by fiberoptic jugular bulb oximetry and changes of intracranial hemodynamics using transcranial Doppler sonography (TCD) during cardiopulmonary bypass (CPB) for coronary artery bypass graft (CABG) in 17 ASA III patients. Anesthesia was maintained with fentanyl, midazolam, and continuous infusion of etomidate. Hypothermic CPB (27 degrees C) was managed according to alpha-stat conditions. SjO2 (%) was measured by a fiberoptic catheter (Opticath F 5.5; Abbott Critical Care Systems) placed in the right jugular bulb via the right internal jugular vein. Mean blood flow velocity (Vmean, cm/s) was measured in the middle cerebral artery using a bidirectional 2-MHz TCD system (Transpect, Medasonics). Data were recorded continuously from the beginning to the end of the CPB. During cooling and hypothermia (27 degrees C); SjO2 and Vmean did not change compared with values at the start of CPB. However, with the beginning of rewarming, Vmean was increased 65% compared with stable hypothermia (27 degrees C). This increase in Vmean was associated with a 25% decrease in SjO2. Maximum desaturation occurred at a 36 degrees C jugular bulb temperature. During cooling and stable hypothermia, global oxygen balance and intracerebral perfusion seemed to be maintained. However, a major alteration in the balance of the cerebral oxygen supply and demand may occur in response to rewarming despite increases in Vmean. Findings suggest inadequate increases in CBF to meet cerebral metabolic demand. Further investigations need to validate these findings with biochemical techniques and neuropsychological tests.


Subject(s)
Brain/blood supply , Cardiopulmonary Bypass , Cerebral Arteries/physiopathology , Coronary Artery Bypass , Oxygen/blood , Anesthesia, General , Blood Flow Velocity , Body Temperature , Cerebral Arteries/diagnostic imaging , Cerebrovascular Circulation , Etomidate , Female , Fentanyl , Humans , Hypothermia, Induced , Intraoperative Period , Jugular Veins , Male , Midazolam , Middle Aged , Oximetry , Ultrasonography, Doppler, Transcranial
15.
Curr Top Microbiol Immunol ; 224: 137-46, 1997.
Article in English | MEDLINE | ID: mdl-9308237

ABSTRACT

The Myc protein activates transcription as part of a complex with its partner protein, Max. Myc-transformed cells are also characterised by the loss of expression of a number of genes and this repressive effect of Myc on gene expression may not be mediated by the Myc/Max complex. We recently isolated by two-hybrid cloning a novel zinc-finger protein that associates with the carboxy-terminus of Myc. We have termed this protein Miz-1 (Myc-interacting zinc finger protein). Some of the properties of Miz-1 suggest that it may be involved in gene repression by Myc in vivo.


Subject(s)
DNA-Binding Proteins/metabolism , Proto-Oncogene Proteins c-myc/metabolism , Transcription Factors/metabolism , Animals , Cell Division , Cell Transformation, Neoplastic , Gene Expression Regulation , HeLa Cells , Humans , Kruppel-Like Transcription Factors , Models, Biological , Zinc Fingers
16.
J Basic Microbiol ; 37(1): 23-8, 1997.
Article in English | MEDLINE | ID: mdl-9090124

ABSTRACT

Transcriptional activation studies involving the human oncoprotein and transcription factor Myc and its helix-loop-helix partner protein Max in mammalian cells are critical due to the presence of endogenous Myc and Max proteins. Here we show that co-expression of the human c-myc and max genes from 2micro circle derived high copy number vectors in yeast cells stimulate the transcriptional activation of a LacZ reporter gene fused to the yeast cytochrome-c1 oxidase minimal promoter containing the adenovirus major late promoter element (AMLPE). The exchange of the single Myc binding site in the AMLPE by the two E-box DNA motifs (CACGTG) present in the Myc responsive element of a human Myc target gene (ornithine decarboxylase) in front of a promoter-reporter gene cassette results in a two-fold enhanced beta-galactosidase expression. Low expression of max and high level expression of c-myc at the same time led to a further enhancement of transcriptional activation from this promoter-reporter gene cassette.


Subject(s)
DNA-Binding Proteins/genetics , Gene Expression Regulation, Fungal , Genes, myc , Saccharomyces cerevisiae/genetics , Transcription Factors , Adenoviridae/genetics , Basic Helix-Loop-Helix Leucine Zipper Transcription Factors , Basic-Leucine Zipper Transcription Factors , Blotting, Western , Cloning, Molecular , Electron Transport Complex IV/genetics , Genes, Reporter , Genetic Vectors , Humans , Lac Operon , Ornithine Decarboxylase/genetics , Plasmids , Promoter Regions, Genetic , Repetitive Sequences, Nucleic Acid , Saccharomyces cerevisiae/metabolism , Transcriptional Activation , Transformation, Genetic , beta-Galactosidase/metabolism
17.
Plant J ; 12(5): 981-90, 1997 Nov.
Article in English | MEDLINE | ID: mdl-9418040

ABSTRACT

Mg-protoporphyrin IX chelatase catalyzes insertion of the magnesium ion into protoporphyrin IX, the last common intermediate precursor in chlorophyll and heme biosynthesis, to form Mg-protoporphyrin IX. In Rhodobacter sphaeroides, and Synechocystis, the three open reading frames bchD/chID, bchH/chIH and bchI/chII encode proteins which are required for in vitro Mg-chelatase activity. In higher plants also, three proteins are necessary for the Mg chelation, and genes homologous to bchH and bchI have been isolated previously. In this study, a novel tobacco cDNA sequence homologous to bchD is isolated and initially characterized. Together with the tobacco clones encoding the other two subunits, full-length cDNAs are now available for the first time for all three subunits of one plant species. The CHL D polypeptide deduced from the open reading frame encodes a protein of 758 aa (82.9 kDa) with an amino terminal extension that resembles a plastid transit peptide. Sequence comparison of tobacco CHL D revealed similarities to the D subunit of Rhodobacter and Synechocystis of 44% and 75%. The amino terminal half of CHL D shows significant similarity (46%) to the entire CHL I peptide sequence, indicating a gene duplication from an ancestral gene. The carboxy terminal half seemed to be unique. Both parts of CHL D are linked with a glutamine/asparagine/proline-rich region flanked by a highly acid-rich segment. Protein-protein interaction among the three subunits CHL D, H and I was studied using the yeast two-hybrid system. Physical interaction was demonstrated between CHL D and CHL I indicating that CHL D is part of the Mg-chelatase. Heterodimer formation of CHL H with CHL I or CHL D could not be demonstrated by transactivation of the lacZ reporter gene. Homodimerization of the CHL D subunit was indicated in the more sensitive assay on X-Gal-containing agar plates. In vitro Mg2+ insertion into protoporphyrin IX was demonstrated in protein extracts of yeast strains expressing the three subunits of tobacco Mg-chelatase. The reconstitution of the recombinant enzyme activity required additional ATP.


Subject(s)
Bacterial Proteins/genetics , Lyases/genetics , Nicotiana/enzymology , Plants, Toxic , Amino Acid Sequence , Bacterial Proteins/biosynthesis , Bacterial Proteins/chemistry , Base Sequence , Cloning, Molecular , Cyanobacteria/genetics , Lyases/biosynthesis , Lyases/chemistry , Macromolecular Substances , Molecular Sequence Data , Open Reading Frames , Recombinant Proteins/biosynthesis , Recombinant Proteins/chemistry , Recombinant Proteins/metabolism , Rhodobacter sphaeroides/genetics , Saccharomyces cerevisiae , Sequence Alignment , Sequence Homology, Amino Acid , Nicotiana/genetics
19.
Article in German | MEDLINE | ID: mdl-9063921

ABSTRACT

INTRODUCTION: This study investigates the accuracy of continuous jugular bulb venous oximetry during different conditions of hypothermic cardiopulmonary bypass (CPB) (27 degrees C) for coronary artery bypass graft. METHODS: 38 ASA III patients were studied following Ethical Care Committee approval and informed consent. Patients were anaesthetized with fentanyl, midazolam, continuous infusion of etomidate and pancuronium. Ventilation was performed with oxygen in air. CPB was managed according to alpha-stat conditions under moderate hypothermia (27 degrees C). SjO2 (%) and jugular bulb temperature (degree C) were measured by a fiberoptic thermodilution catheter (Opticath F 5.5, Abbott Critical Care Systems) placed in the jugular bulb via the internal jugular vein. Appropriate catheter position was x-ray controlled prior to the measurements. The fiberoptic data were compared to co-oximetric data of blood samples after induction of anaesthesia, 2 min following start of CPB, during cooling, stable hypothermia and rewarming of CPB. STATISTICS: Assessing of agreement (Bland/Altman and Bartko). RESULTS: Jugular venous oximetry correlated closely with the co-oximeter determinations after induction of anaesthesia. However, following start of CPB accuracy was decreased. During cooling, stable hypothermia and rewarming oximetric data correlated well with co-oximetry, however, over-estimating the SjO2 by 1.4 to 2%. CONCLUSION: The present data show that continuous jugular bulb venous oximetry is accurate and reliable for continuous SjO2 monitoring during hypothermic CPB for cardiac surgery. Induction of CPB and hemodilution affect accuracy slightly, but changes are well detected. Before clinical intervention SjO2 should be confirmed by laboratory co-oximetry.


Subject(s)
Brain/blood supply , Coronary Artery Bypass , Extracorporeal Circulation/instrumentation , Monitoring, Intraoperative/instrumentation , Oximetry/instrumentation , Adult , Aged , Female , Humans , Jugular Veins , Male , Middle Aged , Sensitivity and Specificity
20.
Article in German | MEDLINE | ID: mdl-8767243

ABSTRACT

UNLABELLED: This study concerns the effects of elevated mean arterial blood pressure (MAP) on decreases in jugular bulb oxygen saturation (SjO2) using fiberoptic jugular bulb oximetry and on cerebral blood flow velocity measured by transcranial Doppler sonography (TCD) during cardiopulmonary bypass (CPB) for coronary artery bypass graft (CABG). METHODS: 27 ASA III patients undergoing CABG were studied. Anaesthesia was maintained with fentanyl, midazolam and continuous infusion of etomidate. CPB was managed according to alpha-stat conditions under moderate hypothermia (27 degrees C). SjO2 (%) and jugular bulb temperature were measured using a fiberoptic catheter placed in the right jugular bulb via the right internal jugular vein. TCD recordings of middle cerebral artery mean blood flow velocity (Vmean, cm/s) were taken during the investigation period. Data were recorded continuously before and for 40 min following start of rewarming. In group 1 (n = 17) MAP was kept between 55 and 65 mmHg, in group 2 (n = 10) MAP was maintained above 70 mmHg using norepinephrine infusion during rewarming of CPB. RESULTS: Following rewarming MAP was statistically significant elevated in group 2 compared to group 1. In groups 1 and 2, Vmean was increased and SjO2 was decreased to a similar extent during rewarming of CPB. Decreases in SjO2 below 50% were seen in both groups. CONCLUSION: The present data show decreases in Sjo2 during rewarming regardless to the level of arterial blood pressure (range 55-80 mmHg). This suggests that desaturation during rewarming of CPB is not a function of decreases in MAP since CBF autoregulation appears to be maintained within this pressure range.


Subject(s)
Blood Pressure/physiology , Brain/blood supply , Coronary Artery Bypass , Coronary Disease/surgery , Extracorporeal Circulation , Oxygen/blood , Rewarming , Adult , Aged , Anesthesia Recovery Period , Blood Flow Velocity/physiology , Coronary Disease/physiopathology , Female , Humans , Hypothermia, Induced , Jugular Veins , Male , Middle Aged , Oxygenators, Membrane
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