Assessment of CXC ligand 12-mediated calcium signalling and its regulators in basal-like breast cancer cells.

CXC ligand (L)12 is a chemokine implicated in the migration, invasion and metastasis of cancer cells via interaction with its receptors CXC chemokine receptor (CXCR)4 and CXCR7. In the present study, CXCL12-mediated Ca2+ signalling was compared with two basal-like breast cancer cell lines, MDA-MB-231 and MDA-MB-468, which demonstrate distinct metastatic potential. CXCL12 treatment induced Ca2+ responses in the more metastatic MDA-MB-231 cells but not in the less metastatic MDA-MB-468 cells. Assessment of mRNA levels of CXCL12 receptors and their potential modulators in both cell lines revealed that CXCR4 and CXCR7 levels were increased in MDA-MB-231 cells compared with MDA-MB-468 cells. Cluster of differentiation (CD)24, the negative regulator of CXCL12 responses, demonstrated increased expression in MDA-MB-468 cells compared with MDA-MB-231 cells, and the two cell lines expressed comparable levels of hypoxia-inducible factor (HIF)2α, a CXCR4 regulator. Induction of epithelial-mesenchymal transition (EMT) by epidermal growth factor exhibited opposite effects on CXCR4 mRNA levels compared with hypoxia-induced EMT. Neither EMT inducer exhibited an effect on CXCR7 expression, however hypoxia increased HIF2α expression levels in MDA-MB-468 cells. Analysis of the gene expression profiles of breast tumours revealed that the highest expression levels of CXCR4 and CXCR7 were in the Claudin-Low molecular subtype, which is markedly associated with EMT features.

Oncosis and apoptosis induction by activation of an overexpressed ion channel in breast cancer cells.

The critical role of calcium signalling in processes related to cancer cell proliferation and invasion has seen a focus on pharmacological inhibition of overexpressed ion channels in specific cancer subtypes as a potential therapeutic approach. However, despite the critical role of calcium in cell death pathways, pharmacological activation of overexpressed ion channels has not been extensively evaluated in breast cancer. Here we define the overexpression of transient receptor potential vanilloid 4 (TRPV4) in a subgroup of breast cancers of the basal molecular subtype. We also report that pharmacological activation of TRPV4 with GSK1016790A reduced viability of two basal breast cancer cell lines with pronounced endogenous overexpression of TRPV4, MDA-MB-468 and HCC1569. Pharmacological activation of TRPV4 produced pronounced cell death through two mechanisms: apoptosis and oncosis in MDA-MB-468 cells. Apoptosis was associated with PARP-1 cleavage and oncosis was associated with a rapid decline in intracellular ATP levels, which was a consequence of, rather than the cause of, the intracellular ion increase. TRPV4 activation also resulted in reduced tumour growth in vivo. These studies define a novel therapeutic strategy for breast cancers that overexpress specific calcium permeable plasmalemmal ion channels with available selective pharmacological activators.

Induction of epithelial-mesenchymal transition (EMT) in breast cancer cells is calcium signal dependent.

Signals from the tumor microenvironment trigger cancer cells to adopt an invasive phenotype through epithelial-mesenchymal transition (EMT). Relatively little is known regarding key signal transduction pathways that serve as cytosolic bridges between cell surface receptors and nuclear transcription factors to induce EMT. A better understanding of these early EMT events may identify potential targets for the control of metastasis. One rapid intracellular signaling pathway that has not yet been explored during EMT induction is calcium. Here we show that stimuli used to induce EMT produce a transient increase in cytosolic calcium levels in human breast cancer cells. Attenuation of the calcium signal by intracellular calcium chelation significantly reduced epidermal growth factor (EGF)- and hypoxia-induced EMT. Intracellular calcium chelation also inhibited EGF-induced activation of signal transducer and activator of transcription 3 (STAT3), while preserving other signal transduction pathways such as Akt and extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation. To identify calcium-permeable channels that may regulate EMT induction in breast cancer cells, we performed a targeted siRNA-based screen. We found that transient receptor potential-melastatin-like 7 (TRPM7) channel expression regulated EGF-induced STAT3 phosphorylation and expression of the EMT marker vimentin. Although intracellular calcium chelation almost completely blocked the induction of many EMT markers, including vimentin, Twist and N-cadherin, the effect of TRPM7 silencing was specific for vimentin protein expression and STAT3 phosphorylation. These results indicate that TRPM7 is a partial regulator of EMT in breast cancer cells, and that other calcium-permeable ion channels are also involved in calcium-dependent EMT induction. In summary, this work establishes an important role for the intracellular calcium signal in the induction of EMT in human breast cancer cells. Manipulation of calcium-signaling pathways controlling EMT induction in cancer cells may therefore be an important therapeutic strategy for preventing metastases.

Hyperbaric oxygen therapy in the treatment of post cardiac surgical strokes--a case series and review of the literature.

Strokes remain an uncommon but significant complication of cardiac surgery. Cerebral air embolism is the likely aetiology in the majority of cases. Hyperbaric oxygen therapy is the recognised treatment for cerebral air embolism associated with compressed air (SCUBA) diving accidents and is therefore also the standard of care for iatrogenic causes of air embolism. It follows that there is a logic in treating post-cardiac surgical stroke patients with hyperbaric oxygen. The aim of this retrospective review was to examine the outcomes of 12 such patients treated in the Christchurch Hospital hyperbaric unit and to appraise the evidence base for the use of hyperbaric oxygen therapy in this setting. Despite delays of up to 48 hours following surgery before the institution of hyperbaric oxygen therapy, 10 of the 12 patients made a full neurological recovery or were left with mild residual symptoms, with nine returning to their previous level of care. One patient remained hemiplegic and there was one early neurological death. There is a paucity of prospective data in this area, but based on sound pathophysiological principles and clinical experience, we believe that patients suffering a stroke following open cardiac surgery should be considered for hyperbaric oxygen therapy.

Impact of plant resistance on southwestern corn borer (Lepidoptera: Crambidae) biology and plant damage.

Southwestern corn borer, Diatraea grandiosella Dyar (Lepidoptera: Crambidae), is a major insect pest of corn, Zea mays L., in the southern United States. Germplasm lines with resistance to southwestern corn borer have been developed and released by the USDA-ARS. Two single-cross hybrids produced by crossing germplasm lines with resistance to southwestern corn borer and a susceptible single-cross hybrid were infested with southwestern corn borer larvae in a 2-yr field test conducted in Mississippi. The susceptible hybrid sustained significantly more leaf damage and stalk tunneling than either resistant hybrid. The number of tunnels and the length of tunneling were significantly lower on the resistant hybrids. In 2003, up to 15 times more tunneling was observed on the susceptible hybrid. Larvae feeding on the resistant hybrids were delayed in their movement from the whorl to the stalk and larval survival was 50% lower on the resistant hybrids than on the susceptible hybrid. Larvae recovered from the susceptible hybrid 7-14 d after infestation weighed twice as much as those recovered from the resistant hybrids. Similar differences in larval weight were observed in the laboratory when larvae were reared on diets prepared from lyophilized tissue from the three hybrids. These results provide a foundation for other investigations designed to identify and determine the roles of specific genes and gene families associated with southwestern corn borer resistance in corn.

Plant resistance and its effect on the peritrophic membrane of southwestern corn borer (Lepidoptera: Crambidae) larvae.

The southwestern corn borer, Diatraea grandiosella Dyar (Lepidoptera: Crambidae), is a serious pest of corn, Zea mays L., in the southern United States. Corn germplasm lines with conventional genetic leaf-feeding resistance to this pest, the fall armyworm, Spodoptera frugiperda (J.E. Smith), and other lepidopterans have been released to the public by USDA-ARS scientists located in Mississippi. Recent studies suggest the insect resistant lines disrupt the integrity of the peritrophic membrane of the fall armyworm. The objectives of this study were to investigate any morphological differences in the structure of the peritrophic membrane of southwestern corn borer larvae feeding on resistant and susceptible corn hybrids and to quantify the damage. Larvae were reared under field and laboratory conditions on three corn hybrids (two resistant and one susceptible). Scanning electron microscopy was used to examine the peritrophic membrane for abnormalities such as holes or tears and to count the holes or tears in the membrane. Differences in the degree of damage to peritrophic membrane of larvae fed on resistant and susceptible plants were not detected. Up to five distinct layers of the membrane were observed in each larva. Variation in the amounts of damage to the peritrophic membrane observed from larvae feeding on all plant material was high. Plant resistance adversely affects growth and development of southwestern corn borer larvae, and further investigations are needed to explain the role of plant resistance and its relation to peritrophic membrane in southwestern corn borer larvae.

Impact of a novel species of Nosema on the southwestern corn borer (Lepidoptera: Crambidae).

A study was undertaken to elucidate the impact of an undescribed Nosema sp. on the southwestern corn borer (SWCB; Diatraea grandiosella Dyar). The Nosema sp. (isolate 506) included in the study was isolated from an overwintering SWCB larva in Mississippi. It was highly infectious per os, with a median infective dose of 2.0 x 10(3) spores per larva. Even at the highest dosage tested (10(7) spores per larva), minimal mortality (< or = 3%) was observed in infected larvae, pupae, and adults reared in the laboratory on an artificial diet. However, infected pupae (0- and 7-d-old) were smaller, and the time to adult eclosion from pupation was slightly increased. Furthermore, the number of eggs produced by infected SWCB female moths substantially decreased (32%), and this effect was most pronounced on day 2, when the greatest number of eggs were oviposited by infected and noninfected moths. For eggs produced by infected females mated with infected males, hatch was slightly decreased by 16 and 15% for eggs laid on days 2 and 3, respectively. In addition, egg hatch was reduced in eggs oviposited by noninfected females mated with infected males on day 3. A low prevalence of infection (< 6%) was observed in the F1 generation originating from infected females mating with noninfected males, from noninfected females mating with infected males, and from infected females mating with infected males. Nosema 506 spores were observed in the proximity of reproductive tissues of infected female and male moths. Spores also were detected on the chorion surface and within eggs laid by infected females. Furthermore, 1-11% of larvae hatching from surface-sterilized eggs were infected by Nosema 506 indicating a transovarial mechanism of transmission.

Impact of Bt cottons expressing one or two insecticidal proteins of Bacillus thuringiensis Berliner on growth and survival of noctuid (Lepidoptera) larvae.

A series of laboratory assays were performed to compare the relative impact of commercial and experimental cultivars of cotton, Gossypium hirsutum (L.), expressing zero, one, or two insecticidal proteins of Bacillus thuringiensis Berliner, on several lepidopteran pests. Assays in which larvae were fed fresh plant tissue indicated that dual-toxin B. thuringiensis (Bt) cultivars, expressing both Cry1Ac and Cry2Ab endotoxins of B. thuringiensis, were more toxic to bollworms, Helicoverpa zea (Boddie), fall armyworms, Spodoptera frugiperda (J. E. Smith), and beet armyworms, Spodoptera exigua (Hubner), than single-toxin cultivars expressing Cry1Ac. Assays in which lyophilized plant tissue was incorporated into artificial diet also indicated improved activity of the dual-toxin Bt cultivar compared with single-toxin plants. Both bollworm and tobacco budworm, Heliothis virescens (F.), growth was reduced by Bt cotton, particularly the dual-toxin cultivar. Although assays with lyophilized tissues were done using largely sublethal doses, bollworm survival was reduced by the dual-toxin cultivar. It appears that this newly developed Bt cotton expressing two toxins will be more effective and have a wider range of activity on these lepidopteran pests.

Humanized anti-IgE mAb Hu-901 prevents the activation of allergen-specific T cells.

BACKGROUND: As a result of the very efficient capture of allergens by IgE that focuses to CD23 on B cells or FcepsilonRI on dendritic cells, allergen-specific T cells can be activated after exposure to very low levels of allergens. This IgE-mediated allergen presentation is 100- to 1,000-fold more efficient than fluid phase endocytosis. The aim of the present study was to determine whether humanized anti-IgE mAb Hu-901 can prevent the activation of allergen-specific T cells by inhibiting IgE-mediated allergen presentation. METHODS: A house dust mite major allergen Der p 1-specific T cell line was generated from an allergic asthma patient, and a model was set up to show IgE-facilitated allergen presentation via CD23 on EBV-transformed B cells. In addition, experiments were performed by FACS analysis, detecting the presence of IgE-allergen complexes bound to EBV-B cells by polyclonal FITC-labeled anti-IgE antisera. RESULTS: The anti-IgE mAb Hu-901 inhibited proliferation of allergen-specific T cells at low allergen concentrations. Inhibition was dose-dependent. This effect could be explained by Hu-901 inhibition of binding of allergen-IgE complexes to CD23 expressed on EBV-transformed B lymphocytes. CONCLUSIONS: These data clearly indicate that anti-IgE antibodies for the treatment of allergy exert their effect not only by inhibiting mast cell/basophil degranulation, but also by preventing T cell activation, which possibly explains the effect of anti-IgE treatment on late-phase reactions noted in clinical studies.

Conventional resistance of experimental maize lines to corn earworm (Lepidoptera: Noctuidae), fall armyworm (Lepidoptera: Noctuidae), southwestern corn borer (Lepidoptera: Crambidae), and sugarcane borer (Lepidoptera: Crambidae).

Plant resistance is a useful component of integrated pest management for several insects that are economically damaging to maize, Zea mays L. In this study, 15 experimental lines of maize derived from a backcross breeding program were evaluated for resistance to corn earworm, Helicoverpa zea (Boddie); fall armyworm, Spodoptera frugiperda (J. E. Smith); southwestern corn borer, Diatraea grandiosella Dyar; and sugarcane borer, Diatraea saccharalis (F.). Experimental line 100-R-3 was resistant in the field to leaf feeding by fall armyworm and line 116-B-10 was resistant in the field to leaf feeding by fall armyworm and leaf and stalk feeding by southwestern corn borer. When corn earworm larvae were fed field harvested silks from experimental line 81-9-B in the laboratory, their pupal weights were significantly lower than the pupal weights of larvae that were fed silks from the resistant control, Zapalote Chico. Maysin levels lower than those commonly associated with corn earworm resistance were present in the resistant experimental line, 107-8-7, indicating a new basis confers resistance to corn earworm in this line. These resistant experimental lines will provide plant breeders with new sources of resistance to lepidopterous insects for the development of improved maize breeding populations.

A unique 33-kD cysteine proteinase accumulates in response to larval feeding in maize genotypes resistant to fall armyworm and other Lepidoptera.

Plants respond to insect feeding with a number of defense mechanisms. Using maize genotypes derived from Antiquan germ plasm that are resistant to Lepidoptera, we have demonstrated that a unique 33-kD cysteine proteinase accumulates in the whorl in response to larval feeding. The abundance of the proteinase increased dramatically at the site of larval feeding after 1 hr of infestation and continued to accumulate for as long as 7 days. The 33-kD cysteine proteinase was most abundant in the yellow-green portion of the whorl-the normal site of larval feeding and the tissue that has the greatest inhibitory effect on larval growth in bioassays. The proteinase was expressed in response to wounding and was found in senescent leaves. It may be a marker of programmed cell death. The gene coding for the proteinase, mir1, has been transformed into Black Mexican Sweet callus. When larvae were reared on callus expressing the proteinase, their growth was inhibited approximately 60 to 80%. The expression of a cysteine proteinase, instead of a cysteine proteinase inhibitor, may be a novel insect defense mechanism in plants.

Influence of whorl region from resistant and susceptible corn genotypes on fall armyworm (Lepidoptera: Noctuidae) growth and development.

The effect of diets prepared from whorl tissue of resistant and susceptible corn genotypes, Zea mays L., on the larval growth, development, and physiology of fall armyworm, Spodoptera frugiperda (J. E. Smith), was analyzed. Larvae reared on an optimized artificial diet had a higher growth rate and developed faster than those reared on lyophilized whorl tissue from resistant and susceptible genotypes. Larvae reared on the resistant material were smaller and had a longer developmental period. Larvae reared on yellow-green and green whorl sections from resistant plants were significantly smaller than those reared on the same sections of susceptible plants. There was no significant difference in weight when larvae were reared on the yellow whorl regions from either resistant or susceptible lines. Physiological indices were determined for larvae fed resistant and susceptible lyophilized and fresh whorl material. Larvae fed resistant lyophilized material had significantly lower growth rate (GW) and efficiency of conversion of ingested food to body substance (ECI) than those reared on artificial diet or susceptible material. However, there were no significant differences in consumption index (CI), approximate digestibility (AD) and efficiency of conversion of digested food to body substance (ECD) between larvae reared on lyophilized tissue from resistant and susceptible genotypes. Larvae reared on fresh yellow-green whorl sections from resistant plants had significantly lower GW, ECI, and ECD than those reared on susceptible material. In contrast, no significant differences in any of the estimated food consumption and utilization indices were observed between larvae reared on fresh yellow whorl sections from resistant or susceptible plants. These results suggest that some components of whorls from resistant plants, especially the yellow-green region, inhibit food utilization in fall armyworm larvae.

Pathogens associated with southwestern corn borers and southern corn stalk borers (Lepidoptera: Crambidae).

A study was undertaken to isolate entomopathogens of southwestern corn borer, Diatraea grandiosella Dyer, and southern corn stalk borer, Diatraea crambidoides (Grote). Field-collected diapausing larvae of southwestern corn borer (three sites in Mississippi) and southern corn stalk borer (one site in North Carolina), and a laboratory strain of D. grandiosella in the diapause state were maintained in a simulated winter followed by a simulated spring environnent. Few larvae (< or = 6%) collected from any of the field sites died in the winter environment, and most insect mortality (11-25%) occurred after transfer of the larvae to the simulated spring environment. Mortality during the simulated spring period differed among the collection sites, and the highest mortality was recorded for southwestern corn borers from Washington County (25%), followed by Marshall (16%) and Oktibbeha (11%) Counties. A high level of mortality was also observed in southern corn stalk borers during the simulated spring period (27%). No viruses were observed, but a number of bacteria, microsporidia, and fungi were isolated from both southwestern corn borer and southern corn stalk borer larvae and pupae. In most instances, numerous bacterial taxa were isolated from cadavers, but on some occasions a single taxon predominated. The most prevalent bacterial taxon from larval and pupal cadavers was Enterococcus faecalis (Andrewes & Horder) Schleifer & Kilppel-Balz, but Bacillus spp., Pseudomonas aeruginosa (Schroeter) Migula, and Serratia marcescens Bizio were frequently isolated as well. Few fungi (1-7%) were recovered from southwestern corn borer and southern corn stalk borer larvae and pupae. The most common entomopathogenic taxon isolated was Beauveria bassiana (Balsamo) Vuillemin from southern corn stalk borer larvae. Microsporidia were not isolated from southern corn stalk borers. However, Nosema spp. were isolated from southwestern corn borer cadavers from Washington (15%), Marshall (1%), and Oktibbeha (3%) Counties in Mississippi. In addition, we observed parasitism of southern corn stalk borer larvae by Macrocentrus cingulum Reinhard (Hymenoptera: Braconidae). No parasitism of southwestern corn borers was observed. Isolates of Bacillus, Beauveria, Entercoccus, Nosema, Pseudomonas and Serratia were all pathogenic to southwestern corn borer larvae under controlled environmental conditions, and with the exception of B. bassiana, these are novel pathogens of Diatraea corn borers.

Subsequent general anaesthesia in patients with a history of previous anaphylactoid/anaphylactic reaction to muscle relaxant.

Of 151 patients with a possible anaphylactoid/anaphylactic reaction to a muscle relaxant investigated over a 20-year period, follow-up for any subsequent general anaesthesia was complete in 145 (96%). One hundred and twenty-two anaesthetics in 72 patients were documented. There were no anaesthetic-related deaths. No subsequent reactions were seen if muscle relaxants were not used in the subsequent anaesthetic, nor were they in patients with severe reactions if the original intradermal test had been equivocal or negative. In the patients with a severe reaction and a positive intradermal test to one or more muscle relaxants, six out of 40 later anaesthetics using muscle relaxants were associated with clinical problems, three being probable anaphylactic reactions, whilst three were minor. Intradermal testing should be performed prior to surgery in this group of patients for the muscle relaxant(s) planned, or an anaesthetic technique which avoids relaxants should be used. This review should encourage other centres to undertake similar follow-up.

Effects of the Southwestern Corn Borer on Aspergillus flavus Kernel Infection and Aflatoxin Accumulation in Maize Hybrids.

Field studies were conducted in 1995 to 1997 to determine the effect of the southwestern corn borer (SWCB) on Aspergillus flavus kernel infection and aflatoxin accumulation in maize hybrids. In 1995, when A. flavus conidia were applied to silks in a spray and SWCB neonate larvae in maize cob grits were placed in the leaf axil at the top ear of commercial hybrids, aflatoxin contamination and A. flavus kernel infection were highest in plants treated with both the fungus and the insect. In 1996, using the same inoculation and infestation techniques, aflatoxin levels and kernel infection were much lower than in 1995 and SWCB had no effect on aflatoxin contamination or kernel infection. In another study in 1996, the effect of SWCB on aflatoxin contamination and A. flavus kernel infection in hybrids resistant and susceptible to A. flavus was determined. The inoculation-infestation technique involved applying maize cob grits containing A. flavus conidia and SWCB larvae to silks. When SWCB was combined with A. flavus, aflatoxin levels and kernel infection were dramatically higher than in hybrids inoculated with A. flavus alone, regardless of whether the hybrids were resistant or susceptible to A. flavus. In 1997, the interaction of A. flavus and SWCB was determined on hybrids resistant and susceptible to A. flavus and on a commercial hybrid with and without the Bacillus thuringiensis (Bt) toxin. Maize cob grits were used to inoculate A. flavus and infest SWCB on the silks 7 or 21 days after midsilk (50% of the plants in a row had silks emerged). All four hybrids had the highest levels of Aspergillus spp. kernel infection and aflatoxin contamination when A. flavus and SWCB were applied at 21 days after midsilk. These studies indicate that SWCB can substantially increase aflatoxin levels when combined with A. flavus. However, inoculation and infestation techniques, placement of the fungus and the insect, and timing of inoculation and infestation are all critical in demonstrating a synergistic relationship between A. flavus and SWCB on aflatoxin contamination of maize.

In vitro regulation of FcepsilonRIalpha expression on human basophils by IgE antibody.

In vivo studies suggested the possibility of an IgE-dependent regulation of high-affinity (FcepsilonRI) IgE receptor expression on basophils. The current studies extend these observations to in vitro cultures of human basophils. Incubation of basophils for 3 to 4 weeks resulted in a slow dissociation of IgE antibody, during which time FcepsilonRI expression decreased, as measured by flow cytometry using the anti-FcepsilonRIalpha monoclonal antibody, 22E7, or by measuring FcRIalpha mass by Western blotting of whole-cell lysates. Culture of basophils with IgE resulted in upregulation of FcepsilonRIalpha expression by both flow cytometry and Western blotting of whole-cell lysates. Upregulation followed a linear time course during 2 weeks of culture. The relative increase in FcepsilonRIalpha density depended on the starting density; with starting densities of FcepsilonRIalpha of 10,000 to 170,000 per basophil, the upregulation varied 20- to 1.1-fold, respectively. Upregulation occurred in high-purity basophils, was not influenced by IgG at concentrations up to 1 mg/mL, and was inhibited by dimeric IgE. Heat-inactivated IgE was less effective and the monoclonal antibody CGP51901 that prevents IgE binding to FcepsilonRIalpha blocked the ability of IgE to induce upregulation. The dose-response curve for IgE-induced upregulation had an effective concentration50 of 230 ng/mL. Although the receptor through which IgE induces this upregulation is not yet known, several characteristics suggest that the upregulation is mediated by IgE interacting through FcepsilonRIalpha itself.

Dermal and airway responses to monoclonal antibodies specific for canine IgE.

In order to understand mechanisms underlying variability of IgE-mediated responses in vivo, we compared effects of different monoclonal antibodies of IgE on dermal and airway responses in a group of atopic dogs. Using intradermal testing, fourteen antibodies were screened in Basenji-Greyhound dogs. For further comparisons between dermal and airway responses, we selected the two antibodies that stimulated the greatest and least dermal responses, respectively. These antibodies bound to IgE with similar affinities (4.1 +/- 0.2 x 10(9) and 1.5 +/- 0.2 x 10(10) M-1). Dose-response curves to intradermal testing were constructed for these two antibodies. On a separate occasion, peripheral airway resistance (Rp) was determined before and after aerosol challenge with an antibody or saline in the same dogs. For one antibody (affinity 4.1 +/- 0.2 x 10(9) M-1), Rp reached a maximum (407 +/- 142% above baseline; mean +/- SE, n = 6) 10 to 15 min after challenge, while maximum responses to saline (62 +/- 16% above baseline, p < 0.01) occurred immediately after aerosol delivery. Responses to the other antibody were similar (p = 0.068) to responses to saline. The magnitude of skin responses did not predict the magnitude of airway responses. These findings suggest that differences in affinities, alone, do not predict magnitude of responsiveness to the anti-IgE antibody and that mechanisms underlying skin and airway responses may differ qualitatively and/or quantitatively.

Efficacy, pharmacodynamics, and pharmacokinetics of CGP 51901, an anti-immunoglobulin E chimeric monoclonal antibody, in patients with seasonal allergic rhinitis.

The efficacy, pharmacodynamics, and pharmacokinetics of CGP 51901, a recombinant monoclonal mouse-human chimeric anti-human immunoglobulin E (IgE) antibody were evaluated for 153 patients with seasonal allergic rhinitis treated with placebo or with 15, 30, or 60 mg CGP 51901 in six biweekly doses. Seasonal allergic rhinitis was chosen to validate the concept of anti-IgE therapy because the causal and temporal relation between allergen confrontation and IgE-mediated evocation of symptoms is firmly established. A sustained 85% or greater reduction of serum free IgE levels was shown to be effective in improving clinical symptoms. The concentration of CGP 51901 needed to maintain 85% or greater reduction of IgE was estimated to be about 5000 ng/ml. Baseline IgE levels and body weights of the patients greatly influenced the pharmacokinetic and pharmacodynamic profiles of CGP 51901. A population model was developed and refined to take into account patient baseline IgE level and body weight. The model was able to help predict multiple-dose pharmacokinetic and pharmacodynamic profiles on the basis of single-dose pharmacokinetic and pharmacodynamic measurements in the therapeutically effective dose range.

Physical aspects of total-body irradiation at the Middlesex Hospital (UCL group of hospitals), London 1988-1993: I. Phantom measurements and planning methods.

This paper, which is divided into parts I and II, describes the physical aspects of work on total-body irradiation (TBI) at the Middlesex Hospital, London, from 1988 to 1993. Irradiation is fractionated and bi-lateral with horizontal accelerator photon beams of 8 MV (1988-1992) at a source-surface distance (SSD) of 3.36 m and 10 MV (1992-1993) at an SSD of 4.62 m. The main aims were maximum patient comfort, a simple, accurate set-up with overall times per fraction of 30 min or less, dose homogeneity throughout the body within +/- 10 to +/- 15%, pre-irradiation treatment planning on nine CT slices using our commercial IGE RTplan (1988-1992) and Target 2 (1992-1993) treatment planning systems and, most important, verification of the plans by in vivo dosimetry to within +/- 5%. Verification of the planned lung doses, which are distributed over five CT slices, was given special attention. In part I of this paper we describe the preliminary work, most of which was done prior to patient treatment. This consisted of standard dosimetric measurements (central axis depth doses, beam profiles at several depths, build-up and build-down curves, beam output calibrations, effect of body compensators, etc), in evaluating silicon diode dosimeters for in vivo dosimetry and of adapting and verifying the methods of treatment planning for TBI conditions. The results obtained with phantoms, including a Rando body phantom, showed that, in principle, our aims could be achieved. The final proof depended, however, on an analysis of the results of the in vivo work and this forms the subject of part II of this paper.

Physical aspects of total-body irradiation at the Middlesex Hospital (UCL group of hospitals), London 1988-1993: II. In vivo planning and dosimetry.

Part II of this paper gives the results of applying the TBI methods described in part I, to in vivo patient planning and dosimetry. Patients are planned on nine CT based body slices, five of which pass through the lungs. Planned doses are verified with ten silicon diodes applied bi-laterally to five body sites, at each treatment. LiF TLDs are applied to seven other body sites at the first treatment only. For 84 patients and at least 1016 measurements per body site with the diodes, the mean measured total doses agreed with planned doses within at most 2% except at lung levels, where the mean measured dose was 3% too low. Standard deviations of the measurements about the mean were between 2.4 and 3.1%. For the LiF TLDs, the mean measured doses for all seven body sites were with in +/- 5% of planned doses. A separate assessment of measured entrance and transmitted doses showed that the former agreed well with planned doses, but that the latter tended to be low, especially over the lungs, and that they had a wider dispersion. Possible reasons for this are discussed. These results show measurement uncertainties similar to those for non-TBI treatments of Nilsson et al, Leunens et al and Essers et al. An analysis of the treatment plans showed a mean dose inhomogeneity in the body (75 patients, nine slices) of 19 +/- 6.0% (1 s.d.) and in the lungs (40 patients, five slices) of 9.2 +/- 2.85% (1 s.d.). The conclusions are that, overall, the methods are reasonably satisfactory but that, with an extra effort, even closer agreement between measured and planned doses and a further limited reduction in the body dose inhomogeneity could be obtained. However, if it were thought desirable to make a substantial reduction in the dose inhomogeneity in the body and lungs, this could only be achieved with the available equipment by changing from lateral to anterior-posterior irradiation and any potential advantages of this change would have to be balanced against a likely deterioration in patient comfort and an increase in treatment set-up times.