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Induction and regulation of xenobiotic-metabolizing cytochrome P450s in the human A549 lung adenocarcinoma cell line.

Several cytochrome P450 (CYP) enzymes are expressed in the human lung, where they participate in metabolic inactivation and activation of numerous exogenous and endogenous compounds. In this study, the expression pattern of all known xenobiotic-metabolizing CYP genes was characterized in the human alveolar type II cell-derived A549 adenocarcinoma cell line using qualitative reverse transcriptase/polymerase chain reaction (RT-PCR). In addition, the mechanisms of induction by chemicals of members in the CYP1 and CYP3A subfamilies were assessed by quantitative RT-PCR. The expression of messenger RNAs (mRNAs) of CYPs 1A1, 1B1, 2B6, 2C, 2E1, 3A5, and 3A7 was detected in the A549 cells. The amounts of mRNAs of CYPs 1A2, 2A6, 2A7, 2A13, 2F1, 3A4, and 4B1 were below the limit of detection. 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induced CYP1A1 and CYP1B1 mRNAs 56-fold and 2.5-fold, respectively. CYP3A5 was induced 8-fold by dexamethasone and 11-fold by phenobarbital. CYP3A4 was not induced by any of the typical CYP3A4 inducers used. The tyrosine kinase inhibitor genistein and the protein kinase C inhibitor staurosporine blocked TCDD-elicited induction of CYP1A1, but they did not affect CYP1B1 induction. Protein phosphatase inhibitors okadaic acid and calyculin A enhanced TCDD-induction of CYP1B1 slightly, but had negligible effects on CYP1A1 induction. These results suggest that CYP1A1 and CYP1B1 are differentially regulated in human pulmonary epithelial cells and give the first indication of the induction of CYP3A5 by glucocorticoids in human lung cells. These results establish that having retained several characteristics of human lung epithelial cell CYP expression, the A549 lung cell line is a valuable model for mechanistic studies on induction of the pulmonary CYP system.

Adenocarcinoma↗

Retinoic acid inhibits induction of c-Jun protein by ultraviolet radiation that occurs subsequent to activation of mitogen-activated protein kinase pathways in human skin in vivo.

Human skin is exposed daily to solar ultraviolet (UV) radiation. UV induces the matrix metalloproteinases collagenase, 92-kD gelatinase, and stromelysin, which degrade skin connective tissue and may contribute to premature skin aging (photoaging). Pretreatment of skin with all-trans retinoic acid (tRA) inhibits UV induction of matrix metalloproteinases. We investigated upstream signal transduction pathways and the mechanism of tRA inhibition of UV induction of matrix metalloproteinases in human skin in vivo. Exposure of human skin in vivo to low doses of UV activated EGF receptors, the GTP-binding regulatory protein p21Ras, and stimulated mitogen-activated protein (MAP) kinases, extracellular signal-regulated kinase (ERK), c-Jun amino-terminal kinase (JNK), and p38. Both JNK and p38 phosphorylated, and thereby activated transcription factors c-Jun and activating transcription factor 2 (ATF-2), which bound to the c-Jun promoter and upregulated c-Jun gene expression. Elevated c-Jun, in association with constitutively expressed c-Fos, formed increased levels of transcription factor activator protein (AP) 1, which is required for transcription of matrix metalloproteinases. Pretreatment of human skin with tRA inhibited UV induction of c-Jun protein and, consequently, AP-1. c-Jun protein inhibition occurred via a posttranscriptional mechanism, since tRA did not inhibit UV induction of c-Jun mRNA. These data demonstrate, for the first time, activation of MAP kinase pathways in humans in vivo, and reveal a novel posttranscriptional mechanism by which tRA antagonizes UV activation of AP-1 by inhibiting c-Jun protein induction. Inhibition of c-Jun induction likely contributes to the previously reported prevention by tRA of UV induction of AP-1-regulated matrix-degrading metalloproteinases in human skin.

Activating Transcription Factor 2↗

Correlation and linear regression between blood pressure decreases after a test dose injection of propofol and that following anaesthesia induction.

Propofol reduces systemic vascular resistance and suppresses cardiac function when injected rapidly. In this study we investigated whether blood pressure decrease after a minimal dose (test-dose) injection of propofol correlates with that after an induction-dose injection. Patients were randomly divided into two groups; anaesthesia was induced in group A (n = 60) using 1.5 mg/kg propofol and in group B (n = 61) using 2.0 mg/kg. Blood pressure reduction after a minimal dose injection (0.4 mg/kg) was examined non-invasively prior to anaesthetic induction. Bispectral Index monitoring was measured and sedation level scored to evaluate anaesthetic depth. After the minimal dose injection, 18 of 121 patients showed behaviour suggesting minor disinhibition, five patients were sedated and seven were drowsy. Oxygen saturation was not significantly changed after test-dose injection. Reduction in systolic blood pressure (mean +/- SD) was 17 +/- 11 mmHg after the minimal dose injection, 42 +/- 20 mmHg after a 1.5 mg/kg induction dose injection, and 42 +/- 22 mmHg after a 2.0 mg/kg induction-dose injection. In both groups, blood pressure after induction was significantly lower than the control value (P < 0.05). In both groups, a positive correlation was observed between blood pressure reduction after the minimal dose injection and that after the induction-dose injection [P < 0.01, R value for systolic blood pressure correlation in group A 0.712 (P < 0.01) and in group B 0.758 (P < 0.01)]. We concluded there was a positive correlation between blood pressure reduction after a minimal (test-dose) injection and that after an induction-dose injection.

Adult↗

The same photoperiodic clock may control induction and maintenance of diapause in the spider mite Tetranchus urticae.

In the spider mite Tetranychus urticae, both diapause induction (which takes place during the larval and nymphal stages) and diapause maintenance (in the adult female) are under photoperiodic control. The question of whether or not the same photoperiodic clock is involved in both photoperiodic reactions was investigated in eight strains of the spider mite, originating from different localities in Europe. The methods employed consisted of (1) determination of the relative importance of the photophase and scotophase in the two photoperiodic reactions; (2) comparison of photoperiodic response curves for diapause induction and diapause maintenance; and (3) determination of the effect of light breaks on the capacity of long nights to maintain diapause, and comparison with the effect of light breaks in diapause induction experiments. The scotophase appeared to be much more important than the photophase for both diapause induction and diapause maintenance. In all strains the critical daylength for diapause maintenance, measured at the moment of saturation of the response to long daylengths, was identical to the critical daylength for diapause induction. However, the critical daylength for diapause maintenance appeared to be labile; it shifted gradually to shorter values as the mites were kept in the cold for a longer period of time, or were kept at a higher temperature for a progressively longer period of time after their stay in the cold room. This seems to reflect a gradual loss of photoperiodic control of diapause maintenance as diapause development proceeds. Photoperiods close to the critical daylength appeared to be less strong with regard to diapause maintenance than shorter daylengths. Quantitative differences in the "strength" of different daylengths were found in all strains investigated. Interruption of the night by short pulses of light revealed either one or two peaks of sensitivity in the night, or one broad "trough" where the two peaks had merged. However, in each case maximal sensitivity to the light breaks occurred at the same position in the night for diapause induction and diapause maintenance. The many similarities found lead to the conclusion that most probably the same photoperiodic clock mechanism is involved in both diapause induction and diapause maintenance in T. urticae.

Animals↗

The impact of TNF-alpha induction on therapeutic efficacy following high dose spatially fractionated (GRID) radiation.

A variety of cytokines especially TNF-alpha and TGF-beta are known to be released in response to high dose radiation of tumors. However, these are not normally measurable in systemic circulation unless high levels of these cytokines are produced by tumor cells. This study was undertaken to see if circulating levels of these cytokines could be measured in the serum of patients treated with high dose spatially fractionated (GRID) radiation and to correlate the finding of these cytokines with clinical response to treatment. Thirty-four patients (31 patients had single treatment site and 3 patients had 2 treatment sites) treated with spatially fractionated (GRID) radiation were entered in this study. Serum samples were collected before treatment and at 24, 48 and 72 hours after GRID radiation. Sandwich enzyme linked immunosorbent assay (ELISA) was performed to estimate the levels of TNF-alpha and activated TGF-beta1 proteins. Seven of 37 patients studied had no TNF-alpha protein before treatment but showed induction of TNF-alpha after GRID radiation. Three patients showed faint basal level of TNF-alpha protein before treatment and these levels were induced after treatment. Three patients showed a basal level of TNF-alpha protein before treatment and these levels decreased after treatment. In 21 cases no TNF-alpha protein was detected before or after treatment at the time points measured. In the case of TGF-beta1 protein, 2 patients showed no TGF-beta1 protein before GRID radiation and an induction of TGF-beta1 protein was observed after treatment. Seven patients showed basal level of TGF-beta1 protein prior to treatment and these levels were induced after treatment. Seventeen patients showed a basal level of TGF-beta1 protein before treatment and these levels decreased after treatment. In 8 cases no TGF-beta1 protein was detected before or after treatment. Complete clinical response (CR) to GRID therapy was seen in 12/37 (32%) treatment sites and partial response (PR) in 18/37 (49%) treatment sites. A strong correlation was observed between clinical CR rate and TNF-alpha induction. The rate of CR was 6/10 (60%) in patients where TNF-alpha was induced as compared to 6/27 (23%) treatment sites in patients where TNF-alpha induction was not seen (p = 0.029). No significant correlation with CR rate and TGF-beta1 induction (44% vs. 28%, p = 0.36) was observed. High dose spatially fractionated (GRID) radiation results in significant induction of TNF-alpha that can be measured in serum of some patients 24 72 hours after radiation. Complete tumor response strongly correlated with the induction of TNF-alpha levels in the serum.

Abdominal Neoplasms↗

A phase 3 study of three induction regimens and of priming with GM-CSF in older adults with acute myeloid leukemia: a trial by the Eastern Cooperative Oncology Group.

The optimal induction for older adults with acute myeloid leukemia (AML) is unknown. Several anthracyclines have been proposed, but the data remain equivocal. Additionally, few prospective trials of priming with hematopoietic growth factors to cycle leukemia cells prior to induction chemotherapy have been conducted. Three hundred and sixty-two older adults with previously untreated AML were randomized to either daunorubicin, idarubicin or mitoxantrone with a standard dose of cytarabine as induction therapy. In addition, 245 patients were also randomized to receive granulocyte-macrophage colony-stimulating factor (GM-CSF) or placebo beginning 2 days prior to induction chemotherapy and continuing until marrow aplasia. No difference was observed in the disease-free overall survival or in toxicity among patients receiving any of the 3 induction regimens or among those receiving growth factor or placebo for priming. However, the complete remission rate for the first 113 analyzable patients, who did not participate in the priming study and started induction therapy 3 to 5 days earlier than those who did, was significantly higher (50% versus 38%; P =.03). None of the anthracyclines is associated with improved outcome in older adults. Priming with hematopoietic growth factor did not improve response when compared with placebo. Furthermore, delaying induction therapy in older adults may lead to a lower complete remission rate.

Aged↗

Effect of airway clearance techniques on the efficacy of the sputum induction procedure.

Sputum induction is used in the early identification of tuberculosis (TB) and pneumocystis infections of the lung. Although manual physiotherapy techniques to clear the airways are often incorporated in the sputum induction procedure, their efficacy in this setting is unknown. This randomised, crossover trial enrolled adults referred for sputum induction for suspected TB and pneumocystis infections of the lung. All participants underwent two sputum induction procedures, inhaling 3% saline via ultrasonic nebuliser. During one randomly allocated procedure, airway clearance techniques (chest wall percussion, vibration, huffing) were incorporated. In total, 59 participants completed the trial. The airway clearance techniques had no significant effect on how the test was tolerated, the volume expectorated or the quality of the sample obtained (assessed by the presence of alveolar macrophages). The techniques did not significantly affect how often the test identified a suspected organism, nor the sensitivity or specificity of sputum induction. In conclusion, the study was unable to demonstrate any effect of airway clearance techniques on the sputum induction procedure. The results provide some justification for not including airway clearance techniques as part of the sputum induction procedure.

Adult↗

An analysis of induction and adjuvant chemotherapy in the multidisciplinary treatment of squamous-cell carcinoma of the head and neck.

This study examines the role of combination chemotherapy with surgery and/or radiotherapy in the initial treatment of patients with advanced stage III and IV squamous-cell carcinoma of the head and neck (SCCHN). Two courses of initial (induction) cisplatin, bleomycin, and methotrexate with oral calcium leucovorin (PBM) were used with the principal intent of increasing the effectiveness of subsequent surgery and/or radiotherapy. Following induction chemotherapy and local treatment, disease-free patients who had responded to initial chemotherapy were entered into a randomized trial of adjuvant PBM. The response rates to induction PBM chemotherapy were a complete response (CR) rate of 26% and a partial response (PR) rate of 52%, for an overall response rate of 78%. A response to induction PBM was highly correlated with failure-free survival (P less than .0001). A Cox multistep regression analysis of potential prognostic factors was performed. After adjusting for the significant prognostic factors of performance status, initial tumor size, and primary tumor site, a response to induction chemotherapy remained independently associated with improved survival (P = .0002). The randomized trial of adjuvant chemotherapy demonstrated that such treatment significantly improved failure-free survival by decreasing local-regional failures. The benefit of adjuvant chemotherapy was particularly evident in patients who had a PR to induction chemotherapy (P = .01). The toxicity of this multidisciplinary approach was predictable and acceptable. Surgery and radiotherapy were not compromised by induction or adjuvant chemotherapy. Definitive evidence that chemotherapy can favorably influence survival awaits confirmation of these results by a randomized trial using a control arm of patients treated with conventional surgery and/or radiotherapy alone.

Aged↗

Whole body 18FDG-PET and the response of esophageal cancer to induction therapy: results of a prospective trial.

PURPOSE: Whole-body 18F-fluorodeoxyglucose ([18F]FDG) positron emission tomography (PET) imaging before and after induction therapy was prospectively evaluated in patients with esophageal cancer to determine whether changes in PET images could measure response to therapy. PATIENTS AND METHODS: Between April 1997 and April 1999, 39 patients (34 men and five women; median age, 59 years; range, 36 to 76 years) with esophageal cancer were prospectively enrolled in a single-institution clinical trial of staging, including PET, induction therapy, restaging including PET, and esophagectomy. All patients undergoing esophagectomy after induction therapy (n = 17) were followed either to recurrence, to death, or through a disease-free interval of at least 24 months. RESULTS: PET after standard staging studies and before therapy imaged undetected sites of metastatic disease in six patients (15%). Restaging (including PET) after induction therapy did not identify any patients with disease progression or any patients with loco-regionally unresectable disease at exploration. The median decrease in the standardized uptake value (SUV) during induction therapy was 59%. After R0 esophagectomy, the 2-year disease-free and overall survival was 38% and 63%, respectively, among patients who had a less than 60% decrease in SUV, and 67% and 89%, respectively, among patients who had a greater than 60% decrease in SUV (P =.055 and P =.088, respectively). CONCLUSION: Compared with conventional imaging, PET detects additional sites of metastatic disease at initial evaluation. After induction therapy, PET did not add to the estimation of loco-regional resectability and did not detect new distant metastases. However, changes in [18F]FDG PET may predict disease-free and overall survival after induction therapy and resection in patients with esophageal cancer. Further evaluation in larger trials is warranted.

Adult↗

Synthetic antiestrogens modulate induction of pS2 and cathepsin-D messenger ribonucleic acid by growth factors and adenosine 3',5'-monophosphate in MCF7 cells.

In MCF7 human breast cancer cells, the antiestrogens 4-hydroxy-tamoxifen and ICI 164,384 inhibit the mitogenic activity of epidermal growth factor (EGF) and insulin-like growth factor-I (IGF-I). These growth factors also stimulate the expression of cathepsin-D and pS2 genes. Therefore, we studied the effects of antiestrogens on growth factor induction of pS2 and cathepsin-D mRNA. The two antiestrogens strongly inhibited the transcriptional induction of pS2 by growth factors. On the contrary, estradiol and IGF-I or EGF had an additive effect on pS2 mRNA accumulation. Growth factor induction of cathepsin-D was also inhibited by ICI 164,384. By contrast, 4-hydroxytamoxifen had an agonist effect on cathepsin-D and an additive effect on IGF-I-induced mRNA. When 12-O-tetradecanoylphorbol-13-acetate or 8-bromo-cAMP (8-Br-cAMP) was used instead of growth factors, similar effects of 4-hydroxytamoxifen and ICI 164,384 were obtained on pS2 (12-O-tetradecanoylphorbol-13-acetate and 8-Br-cAMP) and cathepsin-D (8-Br-cAMP) induction. A mechanism based on the classical competitive inhibition by antiestrogens of estrogen binding and action on the estrogen receptor was very unlikely, as 1) no antigrowth factor activity was obtained with R5020, which was a potent inhibitor of estrogen induction of pS2 and cathepsin-D mRNA; 2) in the Ishikawa endometrial cancer cell line, the cathepsin-D gene is unresponsive to estrogen, but was inhibited by antiestrogen after its induction by EGF or 8-Br-cAMP; and 3) the residual estrogen concentration in cells was too low to induce the expression of estrogen-specific genes. However, antiestrogens did not inhibit the expression of all genes induced by growth factors, as they were without effect on IGF-I induction of glyceraldehyde-3-phosphate dehydrogenase mRNA. These results demonstrate that antiestrogens can modulate the transcription of some growth factor-induced genes and strongly suggest that this effect is not due to interference with residual estrogens.

8-Bromo Cyclic Adenosine Monophosphate↗

Comparison of induction, maintenance, and recovery characteristics of sevoflurane-N2O and propofol-sevoflurane-N2O with propofol-isoflurane-N2O anesthesia.

Induction of, maintenance of, and recovery from sevoflurane anesthesia were compared with propofol and isoflurane anesthesia when administered with nitrous oxide to patients undergoing gynecologic surgery. Seventy-five healthy (ASA I or II), consenting patients were randomly assigned to receive either (I) propofol for induction of anesthesia and isoflurane-nitrous oxide for maintenance (control), (II) propofol for induction and sevoflurane-nitrous oxide for maintenance, or (III) sevoflurane-nitrous oxide for induction and maintenance of anesthesia. Inhaled induction of anesthesia with sevoflurane-nitrous oxide was rapid (109 +/- 25 s to loss of consciousness) and without any untoward hemodynamic changes or episodes of coughing and laryngospasm. Mean arterial blood pressure after induction of anesthesia with propofol (71 +/- 11, 73 +/- 12 mm Hg for groups I and II, respectively) was lower than when sevoflurane (80 +/- 14 mm Hg) was used. The emergence time after discontinuation of isoflurane-nitrous oxide (6.7 +/- 2.2 min) was significantly longer than after propofol-sevoflurane-nitrous oxide or sevoflurane-nitrous oxide alone (4.1 +/- 2.2 and 4.0 +/- 2.0 min for groups II and III, respectively). However, later recovery events did not differ between groups. Serum fluoride levels increased after administration of sevoflurane but not isoflurane. The levels of fluoride ions correlated with the degree of exposure to sevoflurane in MAC-hours. In conclusion, induction of anesthesia with either propofol or sevoflurane-nitrous oxide was rapid and without significant side effects. Emergence and early recovery after maintenance of anesthesia with sevoflurane-nitrous oxide was significantly faster than that after an isoflurane-nitrous oxide combination.

Adult↗

Prevention of atelectasis formation during induction of general anesthesia.

UNLABELLED: General anesthesia promotes atelectasis formation, which is augmented by administration of large oxygen concentrations. We studied the efficacy of positive end-expiratory pressure (PEEP) application during the induction of general anesthesia (fraction of inspired oxygen [FIO(2)] 1.0) to prevent atelectasis. Sixteen adult patients were randomly assigned to one of two groups. Both groups breathed 100% O(2) for 5 min and, after a general anesthesia induction, mechanical ventilation via a face mask with a FIO(2) of 1.0 for another 5 min before endotracheal intubation. Patients in the first group (PEEP group) had continuous positive airway pressure (CPAP) (6 cm H(2)O) and mechanical ventilation via a face mask with a PEEP of 6 cm H(2)O. No CPAP or PEEP was applied in the control group. Atelectasis, determined by computed radiograph tomography, and analysis of blood gases were measured twice: before the beginning of anesthesia and directly after the intubation. There was no difference between groups before the anesthesia induction. After endotracheal intubation, patients in the control group showed an increase of the mean area of atelectasis from 0.8% +/- 0.9% to 4.1% +/- 2.0% (P = 0.0002), whereas the patients of the PEEP group showed no change (0.5% +/- 0.6% versus 0.4% +/- 0.7%). After the intubation with a FIO(2) of 1.0, PaO(2) was significantly higher in the PEEP group than in the control (591 +/- 54 mm Hg versus 457 +/- 99 mm Hg; P = 0.005). Atelectasis formation is prevented by application of PEEP during the anesthesia induction despite the use of large oxygen concentrations, resulting in improved oxygenation. IMPLICATIONS: Application of positive end-expiratory pressure during the induction of general anesthesia prevents atelectasis formation. Furthermore, it improves oxygenation and probably increases the margin of safety before intubation. Therefore, this technique should be considered for all anesthesia induction, at least in patients at risk of difficult airway management during the anesthesia induction.

Adolescent↗

Prevention of atelectasis formation during the induction of general anesthesia in morbidly obese patients.

UNLABELLED: Atelectasis caused by general anesthesia is increased in morbidly obese patients. We have shown that application of positive end-expiratory pressure (PEEP) during the induction of anesthesia prevents atelectasis formation in nonobese patients. We therefore studied the efficacy of PEEP in morbidly obese patients to prevent atelectasis. Twenty-three adult morbidly obese patients (body mass index >35 kg/m(2)) were randomly assigned to one of two groups. In the PEEP group, patients breathed 100% oxygen (5 min) with a continuous positive airway pressure of 10 cm H(2)O and, after the induction, mechanical ventilation via a face mask with a PEEP of 10 cm H(2)O. In the control group, the same induction was applied but without continuous positive airway pressure or PEEP. Atelectasis, determined by computed tomography, and blood gas analysis were measured twice: before the induction and directly after intubation. After endotracheal intubation, patients of the control group showed an increase in the amount of atelectasis, which was much larger than in the PEEP group (10.4% +/- 4.8% in control group versus 1.7% +/- 1.3% in PEEP group; P < 0.001). After intubation with a fraction of inspired oxygen of 1.0, PaO(2) was significantly higher in the PEEP group compared with the control group (457 +/- 130 mm Hg versus 315 +/- 100 mm Hg, respectively; P = 0.035) We conclude that in morbidly obese patients, atelectasis formation is largely prevented by PEEP applied during the anesthetic induction and is associated with a better oxygenation. IMPLICATIONS: Application of positive end-expiratory pressure during induction of general anesthesia in morbidly obese patients prevents atelectasis formation and improves oxygenation. Therefore, this technique should be considered for anesthesia induction in morbidly obese patients.

Adult↗

Acute hemodynamic collapse after induction of general anesthesia for emergent pulmonary embolectomy.

Patients undergoing pulmonary embolectomy often experience hemodynamic deterioration during induction of general anesthesia (GA). Therefore, we studied the incidence and possible risk factors for hemodynamic deterioration during GA induction. Fifty-two consecutive patients undergoing emergent pulmonary embolectomy at our institution were included. Hemodynamic collapse after GA induction was defined as hypotension refractory to vasopressor, inotrope, or fluid administration, requiring cardiopulmonary resuscitation followed by urgent institution of cardiopulmonary bypass (CPB). Demographic variables, comorbidities, specific location of thromboemboli, preoperative inotropic support, and anesthetic drugs used for GA induction were evaluated as possible risk factors. After GA induction, hemodynamic collapse occurred in 19% of patients (n = 10). However, the occurrence of hemodynamic instability was not predicted by any of the evaluated risk factors. In addition, the incidence of in-hospital mortality did not differ between hemodynamically stable or unstable patients (10%; 4 of 42 versus 10%; 1 of 10 patients, respectively). In conclusion, hemodynamic deterioration after GA induction develops frequently during emergent pulmonary embolectomy. On the basis of our experience from this study and the unpredictable nature of hemodynamic deterioration, we suggest that patients undergoing pulmonary embolectomy should be prepared and draped before GA induction, and a cardiac surgical team should immediately be available for emergent institution of cardiopulmonary bypass.

Acute Disease↗

The induction and maintenance of flowering in Impatiens.

The mechanisms that establish the floral meristem are now becoming clearer, but the way in which flowering is maintained is less well understood. Impatiens balsamina provides a unique opportunity to address this question because reversion to vegetative growth can be obtained in a predictable way by transferring plants from inductive to non-inductive conditions. Following increasing amounts of induction, reversion takes place at progressively later stages of flower development. Partial flower induction and defoliation experiments show that a floral signal is produced in the cotyledon in response to inductive conditions and that this signal progressively diminishes after transfer to non-inductive conditions, during reversion. Therefore reversion in Impatiens is most likely due to the failure of leaves to become permanent sources of inductive signal in addition to the lack of meristem commitment to flowering. Analysis of the expression of the Impatiens homologues of the meristem identity genes floricaula and squamosa indicates that a change in floricaula transcription is not associated with the establishment or maintenance of the floral meristem in this species. Squamosa transcription is associated with floral development and petal initiation, and is maintained in existing petal or petaloid primordia even after the meristem has reverted. However, it is not expressed in the reverted meristem, in which leaves are initiated in whorled phyllotaxis and without axillary meristems, both characteristics usually associated with the floral meristem. These observations show that squamosa expression is not needed for the maintenance of these floral characters. The requirement for the production of the floral signal in the leaf during the process of flower development may reflect an additional function separate to that of squamosa activation; alternatively the signal may be required to ensure continued transcriptional activation in the meristem.

Amino Acid Sequence↗

Head induction in the chick by primitive endoderm of mammalian, but not avian origin.

Different types of endoderm, including primitive, definitive and mesendoderm, play a role in the induction and patterning of the vertebrate head. We have studied the formation of the anterior neural plate in chick embryos using the homeobox gene GANF as a marker. GANF is first expressed after mesendoderm ingression from Hensen's node. We found that, after transplantation, neither the avian hypoblast nor the anterior definitive endoderm is capable of GANF induction, whereas the mesendoderm (young head process, prechordal plate) exhibits a strong inductive potential. GANF induction cannot be separated from the formation of a proper neural plate, which requires an intact lower layer and the presence of the prechordal mesendoderm. It is inhibited by BMP4 and promoted by the presence of the BMP antagonist Noggin. In order to investigate the inductive potential of the mammalian visceral endoderm, we used rabbit embryos which, in contrast to mouse embryos, allow the morphological recognition of the prospective anterior pole in the living, pre-primitive-streak embryo. The anterior visceral endoderm from such rabbit embryos induced neuralization and independent, ectopic GANF expression domains in the area pellucida or the area opaca of chick hosts. Thus, the signals for head induction reside in the anterior visceral endoderm of mammals whereas, in birds and amphibia, they reside in the prechordal mesendoderm, indicating a heterochronic shift of the head inductive capacity during the evolution of mammalia.

Animals↗

Adjusting the thermostat: the threshold induction temperature for the heat-shock response in intertidal mussels (genus Mytilus) changes as a function of thermal history.

Spatio-temporal variation in heat-shock gene expression gives organisms the ability to respond to changing thermal environments. The temperature at which heat-shock genes are induced, the threshold induction temperature, varies as a function of the recent thermal history of an organism. To elucidate the mechanism by which this plasticity in gene expression is achieved, we determined heat-shock protein (Hsp) induction threshold temperatures in the intertidal mussel Mytilus trossulus collected from the field in February and again in August. In a separate experiment, threshold induction temperatures, endogenous levels of both the constitutive and inducible isoforms of Hsps from the 70 kDa family and the quantity of ubiquitinated proteins (a measure of cellular protein denaturation) were measured in M. trossulus after either 6 weeks of cold acclimation in the laboratory or acclimatization to warm, summer temperatures in the field over the same period. In addition, we quantified levels of activated heat-shock transcription factor 1 (HSF1) in both groups of mussels (HSF1 inducibly transactivates all classes of Hsp genes). Lastly, we compared the temperature of HSF1 activation with the induction threshold temperature in the congeneric M. californianus. It was found that the threshold induction temperature in M. trossulus was 23 degrees C in February and 28 degrees C in August. This agreed with the acclimation/acclimatization experiment, in which mussels acclimated in seawater tables to a constant temperature of 10-11 degrees C for 6 weeks displayed a threshold induction temperature of 20-23 degrees C compared with 26-29 degrees C for individuals that were experiencing considerably warmer body temperatures in the intertidal zone over the same period. This coincided with a significant increase in the inducible isoform of Hsp70 in warm-acclimatized individuals but no increase in the constitutive isoform or in HSF1. Levels of ubiquitin-conjugated protein were significantly higher in the field mussels than in the laboratory-acclimated individuals. Finally, the temperature of HSF1 activation in M. californianus was found to be approximately 9 degrees C lower than the induction threshold for this species.

Acclimatization↗

Induction of hepatic metallothionein by trivalent cerium: role of interleukin 6.

Metallothionein (MT) is a small sulfydryl-rich protein that binds to and is inducible by heavy metals such as mercury, cadmium, zinc, and copper. However, little is known about the induction of MT by trivalent metals except for bismuth. In this study, we examined the induction of MT synthesis by cerium, a trivalent lanthanoid metal. Administration of cerium chloride (CeCl3) to mice resulted in accumulation of cerium and induction of MT in the liver in a dose-dependent manner. Distribution profiles of metals in the soluble fraction of the liver of CeCl3-treated mice analyzed by high performance liquid chromatography/inductively coupled argon plasma-mass spectrometry (HPLC/ICP-MS) demonstrated that the metal bound to MT-I and MT-II was zinc, but not cerium. Administration of CeCl3 caused increases in the activities of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and the levels of serum amyloid A (SAA), an acute phase protein. Among inflammatory cytokines examined, interleukin 6 (IL-6) exhibited a marked increase in the serum at 3 h after the CeCl3 administration. In order to evaluate the involvement of IL-6 in the induction of MT by cerium, we examined MT induction by CeCl3 in IL-6 null mice. Both the induction of hepatic MT and the increases in SAA levels were markedly suppressed in IL-6 null mice. These results suggest that IL-6 plays an important role in the induction of hepatic MT by cerium.

Animals↗