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At least 19 recordsLinked to original sources
PubMedDate not supplied

Effects of tumor cell-derived interleukin 1 alpha on invasiveness of metastatic clones of murine RCT sarcoma through endothelial cells.

Interleukin 1 alpha (IL-1alpha) production and invasiveness through mouse lung endothelial cells (MLE) were investigated in high-metastatic RCT+ and low-metastatic RCT- clones established from poorly differentiated murine sarcoma. Apparently, a higher level of IL-1alpha was derived from RCT+ cells than from RCT- cells. In an invasion assay, the number of cells which penetrated the MLE monolayer in RCT+ was significantly greater than that in RCT-. The invasiveness of RCT+ and RCT- cells was stimulated by additional recombinant mouse IL-1alpha (rIL-1alpha) in a dose-dependent manner. Anti-mouse IL-1alpha monoclonal antibody (anti-IL-1alpha mAb) significantly inhibited the invasiveness of RCT+ and RCT- cells through the MLE monolayer. However, in RCT+ cells these effects were higher than in RCT- cells. In an attachment assay, the ability of RCT+ cells to attach to the MLE monolayer was significantly higher than that of RCT- cells. The attachment ability of RCT+ and RCT- cells to the MLE monolayer was significantly increased by the pretreatment with rIL-1alpha in a dose-dependent manner. In a retraction assay, conditioned medium of RCT+ stimulated the retraction of the MLE monolayer more markedly in comparison with conditioned medium of RCT-. The retraction of the MLE monolayer was stimulated by additional rIL-1alpha in a dose-dependent manner. The increased retraction of the MLE monolayer was closely associated with the enhancement in tumor cell invasiveness. These findings suggest that IL-1alpha derived from RCT+ and RCT- cells might contribute to the enhancement of tumor cell invasion by stimulating the attachment to the MLE monolayer and retraction of the MLE monolayer.

Animals↗
PubMed1994

Effects of epidermal growth factor on invasiveness through the extracellular matrix in high- and low-metastatic clones of RCT sarcoma in vitro.

We investigated the invasiveness of tumor cells through the extracellular matrix and the influence of epidermal growth factor (EGF) on tumor cell invasion using in vitro systems in high-[RCT(+)] and low-metastatic [RCT(-)] clones established from poorly differentiated murine RCT sarcoma in C3H/He mice. In the invasion assay using a filter coated with reconstituted basement membrane (Matrigel) in a Boyden chamber, RCT(+) cells were more invasive than RCT(-) cells. The attachment of RCT(+) cells to extracellular matrix components and the degradation of type IV collagen by the cells were significantly greater than with RCT(-) cells. However, there was no significant difference in the migration of cells to the extracellular matrix components between cultured RCT(+) and RCT(-) cells. These findings suggested that the different invasiveness of these clone cells was associated with the difference in the ability of attachment to and degradation of the matrix. The level of laminin receptor expression in RCT(+) cells was about four-fold that in RCT(-) cells and laminin stimulated the type IV collagenolytic activity of RCT(+) cells, suggesting that RCT(+) cell attachment to laminin via laminin receptor on the cell surface induced the production of type IV collagenase by the tumor cells. EGF did not affect the invasiveness of RCT(-) cells. In RCT(+) cells, EGF stimulated the invasiveness through Matrigel, the attachment to extracellular matrix components and the degradation of type IV collagen through high-affinity EGF receptors (EGFR), with Kd of pM order, while the migration to the matrix was not influenced by EGF. These findings suggest that the stimulatory effect of EGF on invasion is related to the acceleration of cell adhesion, and the degradative cascade of the extracellular matrix and high-affinity EGFRs play an important role in the effect of EGF on in vitro invasiveness in this tumor.

Animals↗
PubMed1989

Isolation and characterization of low- and high-metastatic clones from murine RCT (Radiological, Chiba, and Toyama) sarcoma.

We have established low- and high-metastatic clones, named RCT(-) and RCT(+) cells, respectively, from the RCT (Radiological, Chiba, and Toyama) sarcoma spontaneously developed in a C3H/He male mouse by the limiting-dilution method in vitro or by the combination of the lung passages and limiting-dilution methods. After 20 serial passages in vitro, the metastatic potential of each clone did not alter. Morphologically, both cells were spindle-shaped, but RCT(+) cells were slightly thicker and larger than RCT(-) cells. The organization of actin-containing filaments was slightly poorer in RCT(+) cells than that in RCT(-) cells. Marked differences were observed in their growth characteristics and adhesiveness to plastic or collagen-coated surfaces, that is, RCT(+) cells grew more slowly but could adhere more rapidly and firmly to the surfaces than RCT(-) cells. RCT(+) cells were agglutinated by all lectins used but several lectins could not agglutinate RCT(-) cells. These results could be a reflection of the difference in oligosaccharide residues on the surface of each cell and, in part, might reflect the difference in organization of the actin-containing filaments that regulate the mobility of lectin receptors. No significant difference between these cell clones was noted in their sensitivity to natural-killer-cell-mediated cytotoxicity in vitro. RCT(-) and RCT(+) cells are considered to be the most useful experimental model for the study of the certain sarcomas.

Animals↗
PubMedDate not supplied

Effects of vascular endothelial growth factor and E-selectin on angiogenesis in the murine metastatic RCT sarcoma.

The relationship between vascular endothelial growth factor (VEGF) and induction of angiogenesis in high-metastatic RCT(+) or low-metastatic RCT(-) clones of the poorly differentiated murine RCT sarcoma was investigated. The association with E-selectin in VEGF-induced angiogenesis was also evaluated. RCT(+) cells produced significantly larger amounts of VEGF than RCT(-) cells. In a tube formation assay with murine lung microvascular endothelial (MLE) cells, conditioned medium from RCT(+) cells showed a significantly greater effect on tube formation than that from RCT(-) cells. Induction of tube formation was suppressed by anti-mouse VEGF monoclonal antibody. Furthermore, anti-mouse E-selectin monoclonal antibody suppressed the tube formation induced by recombinant mouse VEGF. In a flow-cytometric analysis, the expression of E-selectin on MLE cells was upregulated after pretreatment with conditioned medium from RCT(+) and RCT(-) cells. Conditioned medium from RCT(+) cells induced a higher expression of E-selectin compared to medium from RCT(-) cells. Anti-VEGF monoclonal antibody prevented the upregulation of E-selectin by the RCT cell-conditioned medium. These findings suggest that E-selectin plays an important role in the angiogenesis induced by VEGF. VEGF derived from tumor cells may enhance angiogenesis by upregulating the expression of E-selectin on vascular endothelial cells.

Animals↗
PubMed1997

Regulation of gelatinase production and invasiveness by organ-specific fibroblasts in high- and low-metastatic clones from murine RCT sarcoma.

Regulation of gelatinase production, invasiveness and migration activity by organ-specific fibroblasts from embryo, subcutaneous and lung tissues of mice were investigated in high-metastatic RCT+ and low-metastatic RCT- clones established from a poorly differentiated murine sarcoma. In the conditioned media of RCT+ cells, mouse skin fibroblasts (MSF) obtained from the tissue of tumor origin (orthotopic) stimulated the production of the 105-kD gelatinase more than C3H/ 10T1/2 clone 8 (C3H/10 T1/2 CL8) or mouse lung fibroblasts (MLF). In the conditioned media of RCT- cells, however, cocultivation with fibroblasts showed only slight stimulatory effects on the production of the 105-kD gelatinase. In the invasion assay, using a reconstituted basement membrane (matrigel), RCT+ cells cocultivated with MSF showed significantly higher invasiveness than those cocultivated with C3H/10T1/2 CL8 or MLF. However, no significant differences were shown in the invasiveness of RCT- cells in cocultivation with three types of fibroblasts and in cultivation without fibroblasts. There was no significant difference in migration activity between RCT+ and RCT- cells cultivated alone. But in the cocultivation of both clones with MSF, the migration activity of RCT+ cells was significantly higher than that of RCT- cells. These findings suggest that MSF might delineate the difference in characteristics related to the metastatic potential of RCT+ and RCT- cells through regulation by organ-specific factors.

Animals↗
PubMed1989

[Characteristics of high- and low-metastatic clones isolated from murine undifferentiated sarcoma (RCT sarcoma)].

We have established low- and high-metastatic clones from the undifferentiated RCT (Radiological Science-Chiba Cancer Center-Toyama Medical and Pharmaceutical University) sarcoma spontaneously developed in a C3H/He male mouse. Metastatic lesions of RCT sarcoma at the lung were inoculated into the subcutaneous tissue which was then cultured and cloned. Clones potential of low- and high-metastatic were named RCT(-) and RCT(+), respectively. RCT(-) cells were cloned by the limiting dilution method in vitro and RCT(+) cells were cloned by the combination of lung passages and limiting dilution methods. After 20 serial passages in vitro, the metastatic potential of each clone remained unchanged. Significant differences were noted in their growing property and adhesiveness to plastic or collagen-coated surfaces; RCT(+) cells grew more slowly, adhered more rapidly and firmly to the surfaces than those of RCT(-) cells. RCT(+) cells were agglutinated by all types of lectins used in this study, while several types of lectins could not agglutinate RCT(-) cells. These results would indicate that there was the difference in oligosaccharide residues on the surface of each cell. No significant difference between these cell clones in their sensitivity to natural killer cell-mediated cytotoxicity in vitro. We believe that RCT(-) and RCT(+) cells are very useful experimental model to study the mechanism of sarcoma metastasis.

Animals↗
PubMed1988

Relation of H-2 expression on murine RCT(+) sarcoma cells to lung colonization and sensitivity to NK cells.

Murine RCT(+) sarcoma cells were sorted using a fluorescence-activated cell sorter with regard to the expression of H-2 antigens and then an increased H-2-expressing subclone was established, and named RCT(+)H-2+. The experimental metastasis of RCT(+) cells was compared with that of RCT(+)H-2+ cells by counting pulmonary colonies on the 21st day after i.v. inoculation of tumor cells (5-10 x 10(4)/mouse). When mice were inoculated with RCT(+) cells, mean numbers of pulmonary colonies were 2.1(range 0-6), 2.8(range 0-7) using 5 x 10(4) and 1 x 10(5) cells, respectively. On the other hand, in the mice inoculated with RCT(+)H-2+ cells, figures obtained were 7.0(range 4-16), 31.9(range 13-79), using 5 x 10(4) and 1 x 10(5) cells, respectively. The survival rate of RCT(+)H-2+ cells was higher than that of RCT(+) cells, when this was assayed in the early stage after i.v. injection of 51Cr-labeled cells (1 x 10(5) cells/mouse). In addition, RCT(+)H-2+ cells were more resistant than RCT(+) cells to lysis mediated by natural killer cells. These data suggest that an increase in metastatic ability was paralleled by an increase in the H-2 antigen expression and a decrease in sensitivity to the natural killer cells.

Animals↗
PubMed1996

Significance of serum laminin and type IV collagen levels for metastasis in murine RCT sarcoma.

We investigated in vivo the correlation between pulmonary metastasis and serum concentrations of laminin and type IV collagen in high-metastatic RCT(+) and low-metastatic RCT(-) clones established from poorly differentiated RCT sarcoma in C3H/He mice. The in vitro invasiveness of these cell clones through the extracellular matrix was also studied. In the in vitro invasion assay, high-metastatic RCT(+) cells showed significantly more invasiveness than low-metastatic RCT(-) cells. The different invasiveness of these cloned cells was associated with their different ability to attach to and degrade the matrix. In mice inoculated with RCT(+) cells, serum levels of laminin and type IV collagen rose with the increase in the number of pulmonary metastatic nodules. Serum levels of these extracellular matrix components started to rise when pulmonary metastatic nodules were macroscopically observed in the RCT(+) group. In mice bearing RCT(-) cells with lower metastatic ability, there were no significant increases in serum extracellular matrix levels. These findings suggested that serum concentrations of laminin and type IV collagen might be useful markers for the early detection of metastasis.

Animals↗
PubMed2001

Two interactive genes responsible for a new inherited cataract (RCT) in the mouse.

We discovered a mutant mouse, RCT (Rinshoken cataract), with a new congenital cataract in strain SJL/J. The opacity of the lens associated with microphthalmia could be observed visually at 3 to 3.5 months of age. Marked degeneration of the lens, including loss of the fine structure of the lens fibers and swelling of epithelial cells with vacuoles of various sizes in the cortex, but no other defects except photoreceptor degeneration in the retina, was detected. Histological change in the lens was first observed at 2 days after birth. No sex-related differences were detected, and normal phenotypes in the F1 progeny of RCT and normal mice indicated that the cataract was recessive. The chromosomal location of the causative gene was determined by interval mapping by using intersubspecific backcross progeny of RCT and MSM/Ms, an inbred strain from the Japanese wild mouse Mus musculus molossinus. Backcross progeny were divided into three groups according to phenotype: mice (1) with an early-onset cataract, which can be detected visually as in RCT mice, (2) with a late-onset cataract, which can be detected histologically but not visually, and (3) with a normal lens. Three phenotypes were found to be expressed by allele combinations of two recessive genes, rct and mrct (a modifier of rct). The rct locus essential for the onset of the cataract was tightly linked to D4Mit278 on Chromosome (Chr) 4 with no recombination. The mrct locus was closely linked to D5Mit239 (chi2 = 66.3, P << 0.00001) on Chr 5.

Animals↗
PubMed1981

[Clinical significance of RCT in liver diseases (author's transl)].

Radionuclide Computed Tomography (RCT) was performed in 40 cases with various liver diseases. Indication of RCT was determined by the fact that usual liver scintigrams showed or suspected space-occupying lesion. In diffuse hepatocellular disease, RCT added diagnostic informations in 10 out of 17 cases. In these patients, RCT referred decreased uptake area to morphological change such as prominent porta hepatis, widening of both lobes or lateral seg. and medial seg, of the left lobe, and so on. In cases with liver tumor, RCT demonstrated the liver by axial plane, so one could easily or precisely diagnose the location of the mass. RCT performance requires only 30 min, additionally, and no additional injection. We conclude RCT should be performed to the patients whose liver scintigrams show or suspect space-occupying lesion.

Adult↗
PubMed1982

[RCT image study by angular interval of sampling].

A multi-purpose gamma camera system which had 2 detectors of opposed position and was capable of radionuclide computed tomography (RCT) imaging was installed in Division of Nuclear Medicine, Chiba Cancer Center Hospital. In this study, the variance of RCT image due to the angular interval of sampling was analyzed. The full width at half maximum (FWHM) and full width at tenth maximum (FWTM) of point source were almost same for all sampling intervals investigated. The distortion of RCT images with uniform activity source could not be detected even for larger intervals. When the RCT images were obtained using a phantom contained 3 line sources and a bar phantom for RCT which were located at right angle with the plane of reconstruction, the images did not change between 1 degrees and 6 degrees of sampling interval, but for 9 degrees or more the distortion of images occurred explicity. For our system, sampling interval of 2 degrees was predicted from the theorectical calculation considering both resolution and statistical noise. But, results from the phantom studies of present work showed that sampling interval of 6 degrees was sufficient to reconstruct RCT images without distortion, and could be applicable to dynamic RCT images.

Models, Structural↗
PubMed1995

Tumor cell attachment to laminin promotes degradation of the extracellular matrix and cell migration in high-metastatic clone cells of RCT sarcoma in vitro.

We investigated the roles of extracellular matrix proteins, laminin and fibronectin, in promoting invasiveness through the extracellular matrix in high-metastatic [RCT(+)] clone cells established from poorly differentiated murine RCT sarcoma in C3H/He mice. Laminin stimulated the type IV collagenolytic activity of RCT(+) cells. After more than 6 h of incubation, the type IV collagenolysis of the cell-conditioned medium was significantly higher in laminin-treated groups compared with the control. The migration activity of RCT(+) cells was stimulated by laminin. However, fibronectin did not influence the type IV collagenolysis or cell migration in this clone cell. The amino acid sequence YIGSR, which is derived from laminin, inhibited the laminin-mediated cell attachment and the laminin-promoted type IV collagenolysis, as well as cell migration of RCT(+) cells. RGD derived from fibronectin did not influence the cell attachment to laminin or Matrigel in this clone. In the invasion assay employing a Matrigel coated filter in a Boyden chamber, YIGSR showed greater inhibition of invasion through the Matrigel than did RGD with RCT(+) cells. YIGSR might inhibit the promoted-matrix degradation and cell migration in response to the cell attachment to laminin by competing with laminin for binding to cell surface laminin receptor. We suggest that laminin-mediated cell attachment to the extracellular matrix may play a role in promoting the matrix degradation and cell migration during metastatic cascades.

Amino Acid Sequence↗
PubMed2000

Significance of serum type IV collagenolytic activities and gelatinase levels for detection of metastasis in murine RCT sarcoma.

We investigated the usefulness of serum type IV collagenolytic activities and gelatinase levels as diagnostic markers of metastasis in the animal model of spontaneous lung metastasis by FITC-labeled type IV collagen degradation assay and zymographic analyses. High-metastatic RCT(+) and low-metastatic RCT(-) clones were used in the present study. The mean serum type IV collagenolytic activity in the RCT(+) group started to increase from two weeks after hind limb amputation, and was 0.45 and 1.29 unit/ml at three and four weeks. These values were significantly higher than those in the control group (p < 0.01 at three weeks; p < 0.001 at four weeks). A correlation between the number of lung nodules and serum type IV collagenolytic activities in the RCT(+) group was found (r = 0.89, p < 0.001). Zymographic analyses indicated that 105-kD gelatinolytic activities of the RCT(+) group were higher than those of the RCT(-) group at three and four weeks. Thus, type IV collagenolytic activities and serum gelatinase levels might be valuable markers for the detection of metastasis.

Animals↗
PubMedDate not supplied

Attachment and invasion of high- and low-metastatic clones of RCT sarcoma in a three-dimensional culture system.

High- and low-metastatic clones established from poorly differentiated murine sarcoma (RCT sarcoma) spontaneously developed in C3H/He mice were used in this study. Attachment and invasion, which are factors likely related to metastatic ability, of these clones were investigated in a three-dimensional culture system using embryonic chick heart fragments and tumor cell aggregates. The effects of Nocodazole, which interferes with the function of microtubules, and cytochalasin B, which affects the actin assembly in cytoplasmic extensions, on these factors were also examined. Metastatic ability was correlated with attachment and invasiveness of RCT sarcoma. Invasiveness of tumor cells was suppressed by the addition of Nocodazole to the culture medium. Since the disappearance of cell polarity, resulting from the inhibition of tubulin assembly, was found to coincide with the suppression of invasion, directional migration is considered to be involved in the invasion of tumor cells. Cytochalasin B caused a marked decrease in the ratio of attachment of tumor aggregates to embryonic chick heart fragments. The increased ability of invasion and attachment of RCT(+) cells compared with RCT(-) cells seems to contribute to their propensity to metastasize.

Animals↗
PubMed2005

Mothers' attitudes to the randomized controlled trial (RCT): the case of acute lymphoblastic leukaemia (ALL) in children.

OBJECTIVES: Survival rates for childhood cancer have improved substantially partly as a result of national and international randomized clinical trials (RCT). However, the decision for families is complex and emotional. Our aim was to describe the views of mothers of children newly diagnosed with ALL regarding consent to randomized controlled trials. DESIGN: Qualitative interview to explore mothers knowledge, and reasons for involving their child in RCTs. Interviews took place in mothers' homes. PARTICIPANTS: Fifty mothers of children with newly diagnosed ALL (age 4-16 years; mean = 7.4) recruited through research nurses at outpatient appointments. RESULTS: All but three families had consented for their child to be treated in the RCT, although there was wide variation in their understanding of the aims, costs and benefits. Most mothers reported the aim of the trial to compare 'old' and 'new' treatments. CONCLUSION: Despite detailed verbal and written information, mothers were poorly informed about the purpose of the trial, and possibility of side effects. Individual preferences for either standard or new treatment were routinely reported. The data raise questions about the extent to which families give truly informed consent to recruitment of their child to an RCT.

Adolescent↗
PubMed1992

Retinoic acid stimulates transcriptional activity from the alkaline phosphatase promoter in the immortalized rat calvarial cell line, RCT-1.

The immortalized rat calvarial bone cell line RCT-1 responds to treatment with retinoic acid (RA) by increased expression of osteoblast phenotype-related features, including the induction of liver/bone/kidney alkaline phosphatase (ALP) activity. ALP mRNA could not be demonstrated in unstimulated cells, but was first detected in cells treated for 6 h with 1 microM RA. Cycloheximide failed to block the RA induction of ALP mRNA, indicating that de novo protein synthesis was not a requirement for the RA effect and that the ALP gene may be a direct target for RA action. This was confirmed by nuclear run-on assays, which demonstrated a 2.5-fold increase in the abundance of ALP transcripts after 6 h of RA treatment. To determine whether the RA responsiveness was mediated by a specific segment of the ALP promoter, RCT-1 cells were transfected with a series of plasmids containing deletions of the 5'-flanking sequence of the human ALP gene fused to the chloramphenicol acetyl transferase (CAT) gene. CAT activity was measured in cells cultured in the presence of RA or vehicle. All but the smallest construct, which contained 44 basepairs up-stream of the initiation of transcription, were found to mediate a 2- to 3-fold increase in the expression of CAT activity in response to RA. Furthermore, when the region -108 to -45 of the human ALP gene was inserted into the expression vector pBLcat2, in a position immediately up-stream of the herpes simplex virus thymidine kinase promoter, the construct was found to mediate a 2-fold enhancement of CAT activity in response to RA. In gel retardation assays, a major band was present corresponding to the formation of a complex between the 32P-labeled probe containing the -108 to -45 sequence and proteins present in nuclear extracts of RCT-1 cells stimulated for 3 h with RA. These data suggest that the sequence of 64 basepairs (-108 to -45) 5' to the transcription start site is involved in the RA inducibility of the human ALP gene.

Alkaline Phosphatase↗
PubMed1998

Forced expression of hic-5, a senescence-related gene, potentiates a differentiation process of RCT-1 cells induced by retinoic acid.

The hic-5 gene encodes the novel LIM protein which has been implicated in cellular senescence and differentiation processes. Previous studies of rat calvarial cells stimulated to differentiate by addition of retinoic acid (R.A.) showed a four-fold increase in hic-5 expression which preceded an increase in the expression of the differentiation markers, alkaline phosphatase and alpha (I) pro-collagen mRNA. These data suggest involvement of hic-5 in osteoblast differentiation. This hypothesis was further examined using rat calvarial RCT-1 cells containing expression vectors of hic-5. The over-expressing clones showed a decrease in proliferation and displayed the differentiation-related phenotypes such as increased basal levels of alpha (I) collagen mRNA expression and high inducibility by R.A. of alkaline phosphatase activity. Conversely, introduction of hic-5 anti-sense vector leads to the inhibition of alpha (I) collagen mRNA following induction by R.A. These results suggest that hic-5 is one of the regulatory molecules involved in the R.A. induced differentiation process of RCT-1 cells.

Aging↗
PubMed2004

An RCT pilot study to test the effects of intravenous midazolam as a conscious sedation technique for anxious children requiring dental treatment--an alternative to general anaesthesia.

AIM: To add to the evidence base for acceptable and effective paediatric conscious sedation techniques in dental primary care. OBJECTIVES: To compare three conscious sedation techniques for primary care as an alternative to dental general anaesthesia (DGA) in children. To assess the feasibility and practicality of running the trial in general dental practice. To form the basis for sample size calculations and assess scales of measurement. DESIGN: Single centre, randomised control trial (RCT). SETTING: Queensway Anxiety Management Clinic (QAMC). A primary care based general and referral dental practice for the management of anxious patients. SUBJECTS, MATERIALS AND METHODS: Sixty five children too anxious for management with relative analgesia, requiring invasive dental procedure for which dental general anaesthesia (DGA) will be required if an alternative cannot be found. INTERVENTIONS: Group 1 (n = 20) - A combination of inhaled medical air and titrated intravenous midazolam. Group 2 (n = 22) - A combination of inhaled 40% nitrous oxide in oxygen and titrated intravenous midazolam. Group 3 (n = 23) - A combination of an inhaled mixture of 0.3% sevoflurane and 40% nitrous oxide in oxygen with titrated intravenous midazolam. MAIN OUTCOME MEASURES: Successful completion of the intended dental treatment with a child who is co-operative and responsive to verbal commands. RESULTS: Fifty per cent (ten children) successfully completed treatment in Group 1, 73% (16 children) in Group 2 and 83% (19 children) in Group 3. This difference was not significant at a 5% level (chi(2) = 5.53, df = 2, P = 0.07) Of the total of 20 failures, eight children in Group 1 and one child in Group 2 were successfully treated with the addition of sevoflurane and nitrous oxide in oxygen. Only two children required referral to a hospital setting for DGA and the remaining nine children were managed with an alternative conscious sedation technique. CONCLUSION: This pilot shows that intravenous midazolam especially in combination with the addition of inhaled nitrous oxide or sevoflurane and nitrous oxide were promising safe and effective techniques, sufficient to justify progression to a definitive RCT with appropriate methods.

Adolescent↗