First-line Palliative Chemotherapy for Colorectal Cancer: a Population-based Analysis of Delivery and Outcomes in a Single-payer Health System.

AIMS: Clinical practice guidelines recommend palliative chemotherapy for most patients with metastatic colorectal cancer. However, outcomes observed in the real world compared with patients enrolled in clinical trials have not been sufficiently described. The objective of this study was to evaluate the delivery and outcomes of first-line palliative chemotherapy administered to patients with colorectal cancer in routine clinical practice compared with clinical trials. MATERIALS AND METHODS: Using linked health administrative data, we carried out a retrospective population-level cohort study on patients diagnosed with colorectal cancer in Ontario, Canada from 2010 to 2019. Patient, disease and treatment characteristics were summarised. The primary outcome was median overall survival, stratified by treatment prescribed and age. Demographics and outcomes in this real-world population were compared with those from pivotal clinical trials. A multivariable Cox regression model reporting hazard ratios and 95% confidence intervals was used to determine factors associated with survival in patients receiving systemic treatment. RESULTS: We identified 70 987 patients with a new diagnosis of colorectal cancer, of which 4613 received first-line chemotherapy for unresectable locally advanced or metastatic disease and formed the study cohort. Fifty-eight per cent were male and the mean age was 63 years. Most had colon cancer (69%), at least one comorbidity (73%) and lived in an urban location (79%). Less than half (47%) had surgery after diagnosis. The most common regimen prescribed was folinic acid, 5-fluorouracil and irinotecan (FOLFIRI) with bevacizumab or epidermal growth factor receptor inhibitors (EGFRi; n = 2784, 60%). Among all treated patients, the median overall survival was 17.1 months, with survival difference by regimen [median overall survival 18.3 for FOLFIRI with bevacizumab or EGFRi, 19.6 for folinic acid, 5-fluorouracil and oxaliplatin (FOLFOX)/capecitabine, oxaliplatin (XELOX) with bevacizumab or EGFRi, 13.6 for FOLFIRI alone and 7.8 for 5-fluorouracil or capecitabine]. Patients aged >80 years were most likely to have received single-agent 5-fluorouracil or capecitabine, and had inferior overall survival compared with their younger counterparts. Compared with pivotal clinical trials, patients in the real world had inferior overall survival outcomes despite similar demographic characteristics (including age and sex). CONCLUSIONS: In this real-world population-based analysis of patients receiving first-line chemotherapy for unresectable locally advanced or metastatic colorectal cancer, survival outcomes were inferior to those reported in randomised trials despite similarities in age and sex. This information can be used when counselling patients in routine practice about expected outcomes.

Associations with response to Poly(ADP-ribose) Polymerase (PARP) inhibitors in patients with metastatic breast cancer.

PARP inhibitors (PARPi) have modest antitumor activity in patients with advanced breast cancer and mutation in BRCA. It is unclear whether some subgroups derive greater benefit from treatment. MEDLINE and EMBASE were searched from inception to March 2021 to identify trials of PARPi in patients with metastatic breast cancer. Objective response rate (ORR) and clinical benefit rate (CBR) to PARPi were extracted and pooled in a meta-analysis using the Mantel Haenszel random effects model. Meta-regression explored the influence of patient and tumor characteristics on ORR and CBR. For randomized trials, hazard ratio comparing PARPi to control therapy were pooled using inverse variance and random effects. Analysis included 43 studies comprising 2409 patients. Among these, 1798 (75%) patients had BRCA mutations and 1146 (48%) were triple negative. In 10 studies (28%; n = 680 patients), the PARPi was given in combination with platinum-based chemotherapy. Weighted mean ORR was 45%; 64% when combined with platinum vs 37% with PARPi monotherapy (p < 0.001). Previous platinum-based chemotherapy was associated with lower ORR (p = 0.02). Compared to standard chemotherapy, progression-free survival was improved (HR 0.64, p < 0.001), but there was no difference in overall survival (HR 0.87, p = 0.06). There were no differences in ORR or CBR between BRCA1 and BRCA2 mutations. PARPi are more active in combination with platinum than as monotherapy, with lower response if given as monotherapy after platinum exposure. Significant improvements in ORR translated to modest improvement in progression-free, but not overall survival. There was no association between ORR and BRCA mutations.

Relative Yield Loss From Stem Borers (Lepidoptera: Crambidae) and Rice Water Weevil (Coleoptera: Curculionidae) in Rice.

The rice water weevil, Lissorhoptrus oryzophilus Kuschel, is the most important economic pest of rice in the United States with the ability to substantially reduce rice yields. Stem borers, including the Mexican rice borer, Eoreuma loftini (Dyar), are emerging as more consistent pests in Louisiana and Texas, but the relationship between blanked panicles (whiteheads) caused by stem borer feeding and yield loss is not well understood. A series of field trials was conducted in Louisiana from 2019 to 2020 to determine the relative yield loss by manipulating infestations of both pests with selective insecticides. Results indicate losses from L. oryzophilus infestations are greater than those for E. loftini in early planted rice. In contrast, relative yield losses were approximately equal in later planted rice in 2019, in which E. loftini infestations were high. Additive impacts of the root-feeding L. oryzophilus and the stem-feeding E. loftini were not observed in our study. Linear regression revealed that each increase of one whitehead per square meter is associated with a 1.7% loss in yield. Results from ratoon rice suggest that protection from L. oryzophilus in the main crop also improves yield in the second crop. Collectively, results demonstrate the continued need for effective L. oryzophilus management programs to prevent economic losses. Conversely, findings suggest that benefits of E. loftini control may be minimal in many instances. Control of E. loftini is most likely to be economical under conditions where high populations are anticipated such as in late-planted fields or regions that previously had high infestations.

Population Trends of the Sugarcane Borer (Lepidoptera: Crambidae) in Louisiana Sugarcane.

The sugarcane borer, Diatraea saccharalis (F.) (Lepidoptera: Crambidae), is the primary pest of sugarcane, Saccharum spp., in Louisiana. Spring populations are not considered economically damaging, but quantifying infestations can provide an indication of the spatial and temporal character of the damaging summer populations. Statewide surveys quantified the density of sugarcane tillers killed by D. saccharalis (deadhearts) from sugarcane fields across the state in spring from 2003 to 2020. Deadheart density varied greatly among years with a high of 1,318/ha in 2003 to a low of 0/ha in 2018. Linear regressions of the 3-yr rolling average showed declines in spring D. saccharalis populations and the percentage of acreage treated with insecticides over 17 yr. Weather factors including minimum winter temperatures and average spring temperatures were poor predictors of D. saccharalis populations. Only total precipitation in the month of April was positively correlated with numbers of deadhearts per hectare. Results suggest overwintering mortality is not a key factor influencing populations of the first generation of D. saccharalis in Louisiana. Total precipitation in the month of July was positively associated with percentage of treated acreage. Spring deadheart density was directly related to percentage of acreage treated with insecticides during the summer. Quantifying first-generation D. saccharalis populations by recording deadheart density can aid in predicting pest pressure later in the growing season.

Influence of Sorghum Cultivar, Nitrogen Fertilization, and Insecticides on Infestations of the Sugarcane Aphid (Hemiptera: Aphididae) in the Southern United States.

The sugarcane aphid, Melanaphis sacchari Zehntner, is an economically damaging pest of sorghum, Sorghum bicolor (L.), across the southern United States. Field experiments investigated impacts of sorghum cultivar, nitrogen fertilization, and insecticides on M. sacchari infestations and sorghum yields in Louisiana and South Carolina in 2017 and 2018. In South Carolina, M. sacchari densities in unprotected plots peaked on 30-31 July of both years before declining by early- to mid-August. In Louisiana, infestations peaked on 26 and 12 July for 2017 and 2018, respectively, and declined by mid-August. Nitrogen fertilization influenced M. sacchari densities in Louisiana in 2018 with the highest-level infestations recorded from plots that received high N rates. Densities of M. sacchari on susceptible sorghum cultivar, DKS 38-88, were 1.5- to 2.3-fold greater than on DKS 37-07 in both years in Louisiana and in 2018 in South Carolina. Nitrogen fertilization was associated with improved sorghum yields in Louisiana experiments. Sorghum yields across experiments were 2- to 4-fold greater in plots protected with multiple insecticide applications than in unprotected plots. Yield from plots with insecticides sprayed once at currently used action thresholds differed from unprotected plots only in the 2018 Louisiana experiment. Results from these experiments indicate insecticidal protection of susceptible sorghum cultivars remains critical throughout much of the southern United States. Further research is needed to develop integrated management programs that incorporate fertilization manipulation, cultivar resistance, and insecticidal control.

West Indian Canefly (Hemiptera: Delphacidae): An Emerging Pest of Louisiana Sugarcane.

The West Indian canefly, Saccharosydne saccharivora (Westwood) (Hemiptera: Delphacidae), is a sporadic pest of sugarcane in Louisiana which has recently emerged as a more consistent threat with outbreaks occurring in 2012, 2016, 2017, and 2019. Surveys of commercial fields in 2016 revealed that S. saccharivora infestations were present throughout Louisiana sugarcane and populations peaked in mid-June before declining. High minimum winter temperatures are generally associated with S. saccharivora outbreaks. Six insecticide evaluations demonstrated effective control with several insecticides including λ-cyhalothrin, flupyradifurone, acetamiprid, and imidacloprid. In five of the six insecticide trials, S. saccharivora infestations had substantially declined by 21 d after treatment. Effects of insecticidal control of S. saccharivora on sugar yields were detected in one of four small plot trials in which yield data were collected. Linear regression revealed S. saccharivora cumulative insect days in a grid sampling study were inversely associated with sugar yields. Results from these collective experiments suggest impacts on sugar yields are influenced by pest density and infestation duration. Differences were detected in numbers of S. saccharivora nymphs and adults as well as sooty mold coverage among commercial sugarcane cultivars with more than twofold increases in the most susceptible compared to resistant cultivars. The research presented herein documents the impact of S. saccharivora to Louisiana sugarcane and provides important ground work for developing effective pest management strategies. Future research efforts should aim to identify ecological factors influencing population dynamics, varietal preferences, and economic thresholds.

Oviposition Preference and Survival of the Mexican Rice Borer (Lepidoptera: Crambidae) in Bioenergy and Conventional Sugarcane and Sorghum.

Oviposition preference and host suitability of the Mexican rice borer, Eoreuma loftini (Dyar) (Lepidoptera: Crambidae), on bioenergy and conventional cultivars of sugarcane, Saccharum spp., and sorghum, Sorghum spp., were examined in a series of greenhouse experiments. Two energycane cultivars, two sugarcane cultivars, two high-biomass sorghum cultivars, and one sweet sorghum cultivar were assessed at two phenological stages (immature and mature). Mature plants possessed greater availability of dry leaf material compared with immature plants, and all E. loftini eggs were observed exclusively on dry leaves. Oviposition did not vary among host combinations (cultivar by phenological stage); however, eggs per plant and eggs per oviposition event were numerically greater on mature plants than immature plants. In a no-choice experiment, survival from egg to adult did not vary among host combinations, with <2.0% of E. loftini larvae surviving to adulthood. Failed establishment by neonates on plants was 13.4- to 53.9-fold greater than successful establishment across all host combinations. Results from this study suggest that plant physical characteristics continue to play an important role in host selection, but further evaluations will be needed to quantify other characteristics which influence host suitability.

Efficacy of Insect Growth Regulators and Diamide Insecticides for Control of Stem Borers (Lepidoptera: Crambidae) in Sugarcane.

Field experiments assessed control of the stem borers, Diatraea saccharalis (F.) and Eoreuma loftini (Dyar), in sugarcane (Saccharum spp.) with selected insect growth regulators and diamide insecticides. Tebufenozide, novaluron, chlorantraniliprole, and flubendiamide were evaluated for D. saccharalis control in four field experiments in Louisiana between 2009 and 2014. Aerial application of the same insecticides was evaluated for control of E. loftini in commercial sugarcane fields in southern Texas in 2012 and 2015. Stalk injury from D. saccharalis in nontreated plots ranged from 7.4-28.1% bored internodes across the four experiments. All insecticides reduced D. saccharalis injury compared with nontreated plots (39.1-99.4% reduction). Better control was achieved with novaluron, flubendiamide, and chlorantraniliprole than with tebufenozide. Pheromone trap-assisted scouting for E. loftini in Texas sugarcane helped to time insecticide applications. Reduced E. loftini injury was observed in diamide-treated plots (3.6-4.7% bored internodes) compared with nontreated controls (13.1%), but not in novaluron- (6.0%) or tebufenozide-treated (8.3%) plots in the 2012 experiment. Significant E. loftini injury was present prior to the aerial insecticide application in 2015, and differences in overall injury were not detected among treatments. However, chlorantraniliprole reduced injury to the top portion of sugarcane stalks. None of the insecticides improved sugar yields in 2012 or 2015. Results suggest insect growth regulators and diamide insecticides fit well in D. saccharalis management programs in Louisiana sugarcane. These chemistries also have potential to improve control of E. loftini, but more research into application strategies is needed to achieve consistent efficacy.

Monitoring Mexican Rice Borer (Lepidoptera: Crambidae) Populations in Sugarcane and Rice With Conventional and Electronic Pheromone Traps.

The Mexican rice borer, Eoreuma loftini (Dyar), is a major pest of sugarcane, Saccharum spp., rice, Oryza sativa L., and other graminaceous crops in Texas and Louisiana. The ability of conventional and electronic pheromone traps to monitor E. loftini in sugarcane and rice habitats was evaluated in two separate 2-yr field studies. Bucket traps baited with a synthetic female sex pheromone monitored E. loftini populations in commercial sugarcane fields in Calcasieu and Jefferson Davis Parishes throughout the 2014 and 2015 growing seasons. The number of E. loftini captured differed among months, but not between years or parishes. The percentage of E. loftini-injured stalks was greater in 2015 than in 2014, peaking in September of both years. Daily trap capture was correlated with the percentage of injured stalks. Injury from E. loftini in Louisiana sugarcane remained relatively low (<3% bored internodes) in both 2014 and 2015. In a second experiment, electronic traps were compared with conventional pheromone traps for monitoring E. loftini populations in sugarcane and rice habitats in Texas in 2013 and 2015. Performance of earlier electronic trap prototypes in 2013 was inconsistent and less effective than conventional traps. Improved trap design in 2015 resulted in more than threefold greater moth capture in electronic traps than in conventional pheromone traps. Electronic traps demonstrated potential to improve monitoring strategies for this pest and should be evaluated for lepidopterous pests in a variety of cropping systems.

Biology and Management of the Mexican Rice Borer (Lepidoptera: Crambidae) in Rice in the United States.

The Mexican rice borer, Eoreuma loftini (Dyar), is an invasive pest of rice, Oryza sativa L., in the Gulf Coast region of the United States. This pest also damages sugarcane, Saccharum spp. hybrids; corn, Zea mays L.; and sorghum, Sorghum bicolor (L.) Moench, and feeds on weedy noncrop grasses. Multiple aspects of integrated pest management including use of pheromone traps, manipulation of planting dates, harvest cutting height, stubble management, noncrop host management, soil fertility management, host plant resistance, use of insecticides, and biological control have been studied for Mexican rice borer management. However, the current management strategy in rice primarily relies on the use of chlorantraniliprole insecticide seed treatments. This profile addresses Mexican rice borer biology and management in rice in the United States.

El barrenador mexicano del arroz [Eoreuma loftini (Dyar)] es una plaga invasora de arroz (Oryza sativa L.) en la región de la Costa del Golfo de Estados Unidos. Esta plaga también afecta híbridos de caña de azúcar (Saccharum spp.), maíz (Zea mays L.), sorgo [Sorghum bicolor (L.) Moench), y se alimenta de malezas gramíneas. Múltiples aspectos del manejo integrado de plagas incluyendo el uso de trampas con feromonas, la manipulación de las fechas de siembra, la altura del corte durante la cosecha, el manejo de rastrojos, el manejo de hospederos alternos, el manejo de la fertilidad del suelo, la resistencia de la plantas, el uso de insecticidas y el control biológico han sido estudiados para el manejo del barrenador mexicano del arroz. Sin embargo, la estrategia de actual de manejo en arroz se basa principalmente en el tratamiento químico de la semilla con el uso del insecticida clorantraniliprol. Esta revisión se enfoca en la biología y manejo del barrenador mexicano del arroz en arroz en los Estados Unidos.

Yield Response to Mexican Rice Borer (Lepidoptera: Crambidae) Injury in Bioenergy and Conventional Sugarcane and Sorghum.

The Mexican rice borer, Eoreuma loftini (Dyar) (Lepidoptera: Crambidae) is an invasive stem borer of sugarcane, Saccharum spp., and sorghum, Sorghum bicolor (L.), and poses a threat against the production of dedicated bioenergy feedstocks in the U.S. Gulf Coast region. A 2-yr field study was conducted in Jefferson County, TX, to evaluate yield losses associated with E. loftini feeding on bioenergy and conventional cultivars of sugarcane and sorghum under natural and artificially established E. loftini infestations. Bioenergy sugarcane (energycane) 'L 79-1002' and 'Ho 02-113' and sweet sorghum 'M81E' exhibited reduced E. loftini injury; however, these cultivars, along with high-biomass sorghum cultivar 'ES 5140', sustained greater losses in fresh stalk weight. Negative impacts to sucrose concentration from E. loftini injury were greatest in energycane, high-biomass sorghum, and sweet sorghum cultivars. Even under heavy E. loftini infestations, L 79-1002, Ho 02-113, and 'ES 5200' were estimated to produce more ethanol than all other cultivars under suppressed infestations. ES 5200, Ho 02-113, and L 79-1002 hold the greatest potential as dedicated bioenergy crops for production of ethanol in the Gulf Coast region; however, E. loftini management practices will need to be continued to mitigate yield losses.

Expansion of the Mexican Rice Borer (Lepidoptera: Crambidae) into Rice and Sugarcane in Louisiana.

The Mexican rice borer, Eoreuma loftini (Dyar) (Lepidoptera: Crambidae), is an invasive pest of sugarcane, Saccharum spp., rice, Oryza sativa L., and other graminaceous crops in the Gulf Coast region of the United States. Traps baited with E. loftini female sex pheromones were used to document establishment and distribution of E. loftini near sugarcane, rice, and noncrop hosts in seven southwest Louisiana parishes from 2009 to 2013. Additional field surveys documented larval infestations in commercial sugarcane and rice. After its initial detection in 2008, no E. loftini were detected in Louisiana in 2009 and only two adults were captured in 2010. Trapping documented range expansion into Cameron, Beauregard, and Jefferson Davis parishes in 2011 and Allen, Acadia, and Vermilion parishes in 2013. During the course of this study, E. loftini expanded its range eastward into Louisiana 120 km from the Texas border (≈22 km/yr). Surveys of larval infestations provided the first record of E. loftini attacking rice and sugarcane in Louisiana. Infestations of E. loftini in rice planted without insecticidal seed treatments in Calcasieu Parish reached damaging levels.

Improved chemical control for the Mexican rice borer (Lepidoptera Crambidae) in sugarcane: larval exposure, a novel scouting method, and efficacy of a single aerial insecticide application.

A three-treatment aerial application insecticide experiment was conducted in five commercial sugarcane, Saccharum spp., fields in south Texas to evaluate the use of pheromone traps for improving chemical control of the Mexican rice borer, Eoreuma loftini (Dyar), in 2009 and 2010. A threshold of 20 moths/trap/wk was used to initiate monitoring for larval infestations. The percentage of stalks with larvae on plant surfaces was directly related to the number of moths trapped. Reductions in borer injury and adult emergence were detected when a threshold of >5% of stalks with larvae present on plant surfaces was used to trigger insecticide applications. Novaluron provided superior control compared with beta-cyfluthrin; novaluron treated plots were associated with a 14% increase in sugar production. A greenhouse experiment investigating establishment and behavior of E. loftini larvae on two phenological stages of stalkborer resistant, HoCP 85-845, and susceptible, HoCP 00-950, sugarcane cultivars determined that more than half of larvae on HoCP 00-950 and > 25% on HoCP 85-845 tunneled inside leaf mid-ribs within 1 d of eclosion, protected therein from biological and chemical control tactics. Exposure time of larvae averaged < 1 wk for all treatments and was shortest on immature HoCP 00-950 and longest on mature HoCP 85-845. This study shows a short window of vulnerability of E. loftini larvae to insecticide applications, and demonstrates the potential utility of pheromone traps for improving insecticide intervention timing such that a single properly timed application may be all that is required.

High throughput workflow for coacervate formation and characterization in shampoo systems.

Cationic cellulosic polymers find wide utility as benefit agents in shampoo. Deposition of these polymers onto hair has been shown to mend split-ends, improve appearance and wet combing, as well as provide controlled delivery of insoluble actives. The deposition is thought to be enhanced by the formation of a polymer/surfactant complex that phase-separates from the bulk solution upon dilution. A standard characterization method has been developed to characterize the coacervate formation upon dilution, but the test is time and material prohibitive. We have developed a semi-automated high throughput workflow to characterize the coacervate-forming behavior of different shampoo formulations. A procedure that allows testing of real use shampoo dilutions without first formulating a complete shampoo was identified. This procedure was adapted to a Tecan liquid handler by optimizing the parameters for liquid dispensing as well as for mixing. The high throughput workflow enabled preparation and testing of hundreds of formulations with different types and levels of cationic cellulosic polymers and surfactants, and for each formulation a haze diagram was constructed. Optimal formulations and their dilutions that give substantial coacervate formation (determined by haze measurements) were identified. Results from this high throughput workflow were shown to reproduce standard haze and bench-top turbidity measurements, and this workflow has the advantages of using less material and allowing more variables to be tested with significant time savings.

Equilibrium dialysis study and mechanistic implications of coenzyme A binding to acetyl-CoA synthase/carbon monoxide dehydrogenase from Clostridium thermoaceticum.

Parameters for the binding of coenzyme A to acetyl-CoA synthase/carbon monoxide dehydrogenase from Clostridium thermoaceticum were determined by equilibrium dialysis. CoA bound to as-isolated native alpha 2 beta 2 enzyme with KD = 10 +/- 8 microM and n = 0.2 +/- 0.1 moles per alpha beta dimer, where KD is the thermodynamic dissociation constant and n is the number of CoAs bound per alpha beta dimer of the enzyme. The enzyme is heterogeneous; for example, only approximately 30% of alpha subunits contain A-clusters with labile Ni ions (the remainder have nonlabile Ni ions and are nonfunctional). The observed n value suggests that CoA binds only to alpha beta units with Ni-labile A-clusters. The CoA binding properties of enzyme lacking labile Ni was essentially the same, indicating that CoA does not bind directly to the Ni of the A-cluster. This was further evidenced by the observation that bound CoA did not inhibit removal of the labile Ni by 1,10-phenanthroline. CoA did not bind CO-reduced enzyme, and the EPR signal exhibited by the one-electron reduced and CO-bound form of the A-cluster was unaffected by the presence of up to 200 microM CoA. In contrast, CoA did bind Ti(III)-citrate-reduced enzyme (KD = 36 +/- 16 microM, n = 0.16 +/- 0.08). Implications of these results for the mechanism of catalysis are discussed.

Stroke risk factor knowledge in Hispanic and non-Hispanic white women in New Mexico: implications for targeted prevention strategies.

Hispanic women in New Mexico have recently experienced an increase in age-adjusted mortality compared with non-Hispanic white women. Since patients' knowledge of stroke risk factors may affect risk factor control, the present study was undertaken to characterize stroke risk factor understanding in Hispanic and non-Hispanic white women in New Mexico. We administered a stroke risk factor knowledge survey to 215 women hospitalized in Albuquerque, New Mexico. Patients were classified by each of three dichotomous groupings: stroke or nonstroke diagnosis; Hispanic or non-Hispanic white ethnicity; history of cardiovascular risk factors. The frequency of specific item responses was determined for each patient grouping. Two-way analysis of variance was used to determine whether composite knowledge score differed among patient groups. Stress was the attribute most commonly thought to be a risk factor for stroke. Although no ethnic differences were found on composite knowledge score, Hispanic women were significantly less likely to report hypertension as a stroke risk factor than non-Hispanic white women. We suggest that stroke risk factor understanding in Hispanic and non-Hispanic white women in New Mexico is inadequate. Insufficient understanding of the consequences of hypertension, including stroke, may diminish the degree of hypertension control that patients achieve. Further study of the relationship between stroke risk factor understanding and health behavior could enhance prevention efforts.

Induction of bcl-2 expression by phosphorylated CREB proteins during B-cell activation and rescue from apoptosis.

Engagement of surface immunoglobulin on mature B cells leads to rescue from apoptosis and to proliferation. Levels of bcl-2 mRNA and protein increase with cross-linking of surface immunoglobulin. We have located the major positive regulatory region for control of bcl-2 expression in B cells in the 5'-flanking region. The positive region can be divided into an upstream and a downstream regulatory region. The downstream regulatory region contains a cyclic AMP-responsive element (CRE). We show by antibody supershift experiments and UV cross-linking followed by denaturing polyacrylamide gel electrophoresis that both CREB and ATF family members bind to this region in vitro. Mutations of the CRE site that result in loss of CREB binding also lead to loss of functional activity of the bcl-2 promoter in transient-transfection assays. The presence of an active CRE site in the bcl-2 promoter implies that the regulation of bcl-2 expression is linked to a signal transduction pathway in B cells. Treatment of the mature B-cell line BAL-17 with either anti-immunoglobulin M or phorbol 12-myristate 13-acetate leads to an increase in bcl-2 expression that is mediated by the CRE site. Treatment of the more immature B-cell line, Ramos, with phorbol esters rescues the cells from calcium-dependent apoptosis. bcl-2 expression is increased following phorbol ester treatment, and the increased expression is dependent on the CRE site. These stimuli result in phosphorylation of CREB at serine 133. The phosphorylation of CREB that results in activation is mediated by protein kinase C rather than by protein kinase A. Although the CRE site is necessary, optimal induction of bcl-2 expression requires participation of the upstream regulatory element, suggesting that phosphorylation of CREB alters its interaction with the upstream regulatory element. The CRE site in the bcl-2 promoter appears to play a major role in the induction of bcl-2 expression during the activation of mature B cells and during the rescue of immature B cells from apoptosis. It is possible that the CRE site is responsible for induction of bcl-2 expression in other cell types, particularly those in which protein kinase C is involved.

Significant reversal of thyrotoxicosis-associated dilated cardiomyopathy with induction of the euthyroid state.

Thyrotoxicosis rarely precipitates heart failure. Older patients and those with underlying cardiac disease are at greater risk of experiencing this complication of thyrotoxicosis. A 43 year old male who presented with subclinical thyrotoxicosis, atrial fibrillation, and a dilated cardiomyopathy is discussed. There was no evidence of clinically significant underlying cardiac disease. At admission, the patient had an ejection fraction of 25%. Thyrotoxicosis was treated with propylthiouracil. At 14 weeks after hospitalization, the patient had an ejection fraction of 50% with significant reduction in cardiac chamber sizes and left ventricular mass index. He was biochemically euthyroid at that time. After ten months of propylthiouracil therapy, he had progressive improvement in cardiac function and decrease in left ventricular mass index. Thyrotoxicosis-associated cardiomyopathy may reverse significantly with treatment of thyrotoxicosis alone; this complication of thyrotoxicosis should be considered in any young individual with a dilated cardiomyopathy of unknown etiology.

Specific targeting of human hepatocellular carcinoma cells by immunoliposomes in vitro.

The monoclonal antibody AF-20 was raised against the human hepatocellular carcinoma (HCC) cell line FOCUS and binds with high affinity to a rapidly internalized 180-kd homodimeric glycoprotein that is abundantly expressed on the surface of human HCC and other human cancer cell lines. Immunoliposomes were produced by covalently coupling AF-20 to liposomes containing carboxyfluorescein. Interaction of immunoliposomes with various HCC cell lines in vitro was quantitatively assessed by flow cytometry and qualitatively analyzed by fluorescence microscopy. Liposomes bearing an isotype-matched nonrelevant monoclonal antibody (MAb) and cell lines not expressing AF-20 antigen served as controls. AF-20-immunoliposomes specifically bound to HCC and other human cancer cell lines expressing the AF-20 antigen and were rapidly internalized at 37 degrees C. Interaction of AF-20-conjugated liposomes with these cell lines was between 5 and 200 times greater than that of unconjugated liposomes, whereas no difference was observed between control liposomes bearing a nonrelevant antibody and unconjugated liposomes. Specificity of liposome-target cell interaction was confirmed by competitive inhibition assays. Kinetic analysis showed rapid association of AF-20 immunoliposomes with target cells, with saturation conditions being reached after 60 minutes. We conclude that the MAb AF-20 directs highly efficient, specific, and rapid targeting of immunoliposomes to human HCC and other human cancer cell lines in vitro. This targeted liposomal delivery system represents a promising approach for the development of immunotargeted diagnosis and therapy strategies against HCC.