Optimization and validation of a micro-QuEChERS method for phthalates detection in small samples of cetacean blubber.

In this study, an innovative method was developed to detect and quantify phthalates in fresh cetacean blubber. An adaptation of the ammonium formate QuEChERS method was used and adapted as a micro-extraction for small quantities of samples. Significantly, this technique utilized minimal quantities of reagents and salts, with the additional implementation of rigorous Quality Assurance/Quality Control protocols to further reduce background contamination. To ensure the reliability of this method, comprehensive validation procedures were conducted, with a specific focus on two widely studied cetacean species: the common bottlenose dolphin (Tursiops truncatus) and the short-finned pilot whale (Globicephala macrorhynchus). Determination coefficients (R2) for matrix-matched calibration were >0.93 with limits of quantifications (LOQ) of the method in the range of 5-10 ng/g. Mean recovery values were between 40 and 100 %. This novel methodology holds particular relevance for environmental research studies, offering the capability to detect emerging contaminants with minimal sample requirements. This aspect is particularly valuable in investigations that involve free-ranging animals and rely on biopsy sampling. It allows for the assessment of contaminant levels in healthy individuals within wild populations, enhancing our understanding of ecological impacts and potential conservation measures.•A micro-extraction adaptation of the ammonium formate QuEChERS method was developed and applied to a small quantity of fresh cetacean blubber to detect phthalates.•Small quantities of reagents and salts were used, and additional Quality Assurance/ Quality Control procedures were taken to further minimize background contamination.•Method validation was carried out for two cosmopolitan and extensively studied cetacean species: the common bottlenose dolphin (Tursiops truncatus) and the short-finned pilot whale (Globicephala macrorhynchus).

Microplastic ingestion and plastic additive detection in pelagic squid and fish: Implications for bioindicators and plastic tracers in open oceanic food webs.

The ubiquitous presence of microplastics (MPs) in the ocean represents a potential threat to marine organisms, with poorly understood long-term adverse effects, including exposure to plastic additives. The present study investigated the ingestion of MPs in two epipelagic fish species (Trachurus picturatus and Scomber colias) and three pelagic squid species (Loligo vulgaris, Ommastrephes caroli and Sthenoteuthis pteropus) from an open oceanic region of the Northeast Atlantic. Seven phthalate esters (PAEs) were also analysed in the organisms' tissue, and the potential correlation between PAEs concentrations and ingested MPs was investigated. Seventy-two fish and 20 squid specimens were collected and analysed. MPs were found in the digestive tract of all species and in the squid species' gills and ink sacs. The highest occurrence of MPs was in the stomach of S. colias (85 %) and the lowest in the stomach and ink sac of O. caroli and L. vulgaris (12 %). Most of the particles identified (>90 %) were fibres. Among all the ecological and biological factors considered (dietary preferences, season, body size, total weight, liver weight, hepatosomatic index and gastrosomatic index), only gastrosomatic index (GSI) and season were significant predictors of MPs ingestion in fish species, with a greater likelihood of ingestion in the cold season and in specimens with higher GSI values (i.e. higher feeding intensity). Four PAEs (DEP, DIBP, BBP, DEHP) were detected in all the species analysed, with average ∑PAEs concentrations ranging between 10.31 and 30.86 ng/g (wet weight). DIBP was positively correlated with ingested MPs, suggesting this compound might represent a "plastic tracer". This study looks into the problem of MPs ingestion for pelagic species in an open oceanic region, highlighting the most suitable bioindicators and providing essential insights into the factors that may influence ingestion rates. Additionally, the detection of PAEs in all species indicates the need for further research on the contamination sources, the effects of these chemicals on marine organisms, and the potential risks to human health through seafood consumption.

Microplastics reduce microalgal biomass by decreasing single-cell weight: The barrier towards implementation at scale.

Microplastics (MPs) are a widespread environmental threat, especially to aquatic and urban systems. Water quality is vital for biomass production in microalgal-based industries. Here, industrially relevant microalgae Tetraselmis suecica, Scenedesmus armatus, and Nannochloropsis gaditana were exposed to PS- and PE-MPs (polystyrene and polyethylene, respectively - 10-20 µm) contaminated waters (5 and 10 mg/L). Following industrial empirical and ecotoxicological procedures, the production period was established as four days (exponential growth phase). 27-long day experiments were conducted to determine the chronic effects of MPs contamination in microalgal biomass yields. MPs induced different responses in cell density: T. suecica decreased (up to 11 %); S. armatus showed no changes; and N. gaditana increased (up to 6 %). However, all three microalgae exhibited significant decreases in biomass production (up to 24, 48, and 52 %, respectively). S. armatus exposed to PS-MPs and N. gaditana exposed to PE-MPs were the most impacted regarding biomass production. The decrease in biomass yield was due to the reduction in single-cell weight (up to 14, 47, and 43 %), and/or the production of smaller-sized cells (T. suecica). In response to chronic exposure, microalgae showed signs of cell density adaptation. Despite cell density normalizing, biomass production was still reduced compared to biomass production in clean water. Computational modelling highlighted that MPs exposure had a concentration-dependent negative impact on microalgae biomass. The models allow the evaluation of the systematic risks that MPs impose in microalgal-based industries and stimulate actions towards implementing systems to contain/eliminate MPs contamination in the waters used in microalgae production.

Bacterial cellulose biopolymers: The sustainable solution to water-polluting microplastics.

Microplastics (MPs) pollution has become one of our time's most consequential issue. These micropolymeric particles are ubiquitously distributed across all natural and urban ecosystems. Current filtration systems in wastewater treatment plants (WWTPs) rely on non-biodegradable fossil-based polymeric filters whose maintenance procedures are environmentally damaging and unsustainable. Following the need to develop sustainable filtration frameworks for MPs water removal, years of R&D lead to the conception of bacterial cellulose (BC) biopolymers. These bacterial-based naturally secreted polymers display unique features for biotechnological applications, such as straightforward production, large surface areas, nanoporous structures, biodegradability, and utilitarian circularity. Diligently, techniques such as flow cytometry, scanning electron microscopy and fluorescence microscopy were used to evaluate the feasibility and characterise the removal dynamics of highly concentrated MPs-polluted water by BC biopolymers. Results show that BC biopolymers display removal efficiencies of MPs of up to 99%, maintaining high performance for several continuous cycles. The polymer's characterisation showed that MPs were both adsorbed and incorporated in the 3D nanofibrillar network. The use of more economically- and logistics-favourable dried BC biopolymers preserves their physicochemical properties while maintaining high efficiency (93-96%). These polymers exhibited exceptional structural preservation, conserving a high water uptake capacity which drives microparticle retention. In sum, this study provides clear evidence that BC biopolymers are high performing, multifaceted and genuinely sustainable/circular alternatives to synthetic water treatment MPs-removal technologies.

Seasonal variation in microplastics and zooplankton abundances and characteristics: The ecological vulnerability of an oceanic island system.

The ingestion of microplastics (MPs - plastic particles <5 mm) by planktivorous organisms represents a significant threat to marine food webs. To investigate how seasonality might affect plastic intake in oceanic islands' ecosystems, relative abundances and composition of MPs and mesozooplankton samples collected off Madeira Island (NE Atlantic) between February 2019 and January 2020 were analysed. MPs were found in all samples, with fibres accounting for 89 % of the particles. MPs and zooplankton mean abundance was 0.262 items/m3 and 18.137 individuals/m3, respectively. Their monthly variations follow the seasonal fluctuation of environmental parameters, such as currents, chlorophyll-a concentration, sea surface temperature and precipitation intensity. A higher MPs/zooplankton ratio was recorded in the warm season (May-Oct), reaching 0.068 items/individual when considering large-sized particles (1000-5000 µm). This is the first study to assess the seasonal variability of MPs in an oceanic island system providing essential information respecting its ecological impact in pelagic environments.

Lipophilic toxins occurrence in non-traditional invertebrate vectors from North Atlantic Waters (Azores, Madeira, and Morocco): Update on geographical tendencies and new challenges for monitoring routines.

In the last decades, due to monitoring programs and strict legislation poisoning incidents occurrence provoked by ingestion of naturally contaminated marine organisms has decreased. However, climate change and anthropogenic interference contributed to the expansion and establishment of toxic alien species to more temperate ecosystems. In this work, the coasts of Madeira, São Miguel islands and the northwestern Moroccan coast were surveyed for four groups of lipophilic toxins (yessotoxins, azaspiracids, pectenotoxins, and spirolides), searching for new vectors and geographical tendencies. Twenty-four species benthic organisms were screened using UHPLC-MS/MS technique. We report 19 new vectors for these toxins, six of them with commercial interest (P. aspera, P. ordinaria, C. lampas, P. pollicipes, H. tuberculata and P. lividus). Regarding toxin uptake a south-north gradient was detected. This study contributes to the update of monitoring routines and legislation policies, comprising a wider range of vectors, to better serve consumers and ecosystems preservation.

The effect of microplastics pollution in microalgal biomass production: A biochemical study.

Microplastics (MPs) are widely spread throughout aquatic systems and water bodies. Given that water quality is one of the most important parameters in the microalgal-based industry, it is critical to assess the biochemical impact of short- and long-term exposure to MPs pollution. Here, the microalga Phaeodactylum tricornutum was exposed to water contaminated with 0.5 and 50 mg L-1 of polystyrene (PS) and/or polymethyl methacrylate (PMMA). Results show that the microalgal cultures exposed to lower concentrations of PS displayed a growth enhancement of up to 73% in the first stage (days 3-9) of the exponential growth phase. Surprisingly, and despite the fact that long-term exposure to MPs contamination did not impair microalgal growth, a steep decrease in biomass production (of up to 82%) was observed. The production of photosynthetic pigments was shown to be pH-correlated during the full growth cycle, but cell density-independent in later stages of culturing. The extracellular carbohydrates production exhibited a major decrease during long-term exposure. Still, the production of extracellular proteins was not affected by the presence of MPs. This pilot laboratory-scale study shows that the microalgal exposure to water contaminated with MPs disturbs its biochemical equilibrium in a time-dependent manner, decreasing biomass production. Thus, microalgal industry-related consequences derived from the use of MPs-contaminated water are a plausible possibility.

Lobophora (Dictyotales) Species Richness, Ecology and Biogeography Across the North-Eastern Atlantic Archipelagos and Description of Two New Species1.

The brown alga Lobophora (Dictyotales, Phaeophyceae) is an important macroalga in the North-eastern Atlantic archipelagos (i.e., Macaronesia). Notably in the Canaries it can dominate benthic assemblages. While the genus has been the subject of several ecological studies in the Canaries, no study has yet been conducted to assess species-level diversity of Lobophora in Macaronesia. We reassessed the diversity of Lobophora in Macaronesia, reporting the presence of seven species (L. caboverdeana sp. nov., L. canariensis, L. dagamae sp. nov., L. delicata, L. dispersa, L. littlerorum, and L. schneideri). Lobophora spp. from Macaronesia are morphologically and ecologically distinguishable. In the Canaries, L. schneideri dominates the photophilic assemblages from the intertidal to 20-30 m depth. Lobophora dagamae sp. nov. grows in less illuminated shallow habitats, and replaces L. schneideri from 30 to ~80 m. Lobophora canariensis also has a wide vertical distribution, from the intertidal to deep waters, while L. delicata, L. dispersa and L. littlerorum grow in shallow waters. The dominance of species with an upright habit versus prostrate or crustose species may be mediated by the pressure of herbivores. Four species have an amphi-Atlantic distribution: L. littlerorum, L. canariensis, L. delicata, and L. schneideri. Lobophora schneideri and L. delicata are furthermore distributed in the Mediterranean Sea. By sampling a pivotal region in the Atlantic, this study significantly improves our knowledge of Lobophora biogeography in the Atlantic Ocean. Macaronesia constitutes a species-poor region for Lobophora where no diversification events occurred, and a region of overlap between the Greater Caribbean and the Indo-Pacific.

Ecotoxicological and biochemical effects of environmental concentrations of the plastic-bond pollutant dibutyl phthalate on Scenedesmus sp.

Phthalate esters are highly present in aquatic plastic litter, which can interfere with the biological processes in the wildlife. In this work, the commonly found freshwater microalga Scenedesmus sp. was exposed to environmental concentrations (0.02, 1 and 100 µg L-1) and to a higher concentration (500 µg L-1) of dibutyl phthalate (DBP), which is an environmental pollutant. The growth, pH variation, production of photosynthetic pigments, proteins and carbohydrates were evaluated. The main inhibition effect of DBP on the microalgal growth was observed in the first 48 h of the exposure (EC50: 41.88 µg L-1). A reduction in the photosynthetic pigment concentration was observed for the 0.02, 1 and 100 µg L-1 conditions indicating that the DBP downregulated the growth rate and affected the photosynthetic process. A significant increase in protein production was only observed under 500 µg L-1 DBP exposure. The extracellular carbohydrates production slightly decreased with the presence of DBP, with a stronger decrease occurring in the 500 µg L-1 condition. These results highlight the environmental risk evaluation and ecotoxicological effects of DBP on the production of biovaluable compounds by microalgae. The results also emphasize the importance of assessing the consequences of the environmental concentrations exposure as a result of the DBP dose-dependent correlation effects.

From a Basic Microalga and an Acetic Acid Bacterium Cellulose Producer to a Living Symbiotic Biofilm.

Bacterial cellulose (BC) has recently been the subject of a considerable amount of research, not only for its environmentally friendly biosynthesis, but also for its high potential in areas such as biomedicine or biomaterials. A symbiotic relationship between a photosynthetic microalga, Chlamydomonas debaryana, and a cellulose producer bacterium, Komagataeibacter saccharivorans, was established in order to obtain a viable and active biofilm. The effect of the growth media composition ratio on the produced living material was investigated, as well as the microalgae biomass quantity, temperature, and incubation time. The optimal temperature for higher symbiotic biofilm production was 30 °C with an incubation period of 14 days. The high microalgae presence, 0.75% w/v, and 60:40 HS:BG-11 medium (v/v) induced a biofilm microalgae incorporation rate of 85%. The obtained results report, for the first time, a successful symbiotic interaction developed in situ between an alkaline photosynthetic microalga and an acetic acid bacterium. These results are promising and open a new window to BC living biofilm applications in medical fields that have not yet been explored.

Tetrodotoxins Occurrence in Non-Traditional Vectors of the North Atlantic Waters (Portuguese Maritime Territory, and Morocco Coast).

Tetrodotoxin (TTX) is a potent alkaloid typically from tropical ecosystems, but in the last decade its presence has been more pronounced in the temperate waters of the Atlantic. In its last scientific opinion, the European Food Safety Authority (EFSA) stressed the need for data regarding TTX prevalence in European waters. To address EFSA's concerns, benthic organisms such as mollusks, crustaceans, echinoderms and fish with different feeding habits were collected along the Portuguese continental coast, islands (São Miguel, Azores, and Madeira) and the northwestern Moroccan coast. A total of 165 samples were analyzed by ultra high performance liquid chromatography high resolution mass spectrometry (UHPLC-HRMS) and ultra high performance chromatography mass spectrometry (UHPLC-MS/MS). Geographical tendencies were detected as follows, by descending order: S. Miguel Island (Azores), Moroccan coast, Madeira Island and Portuguese continental coast. The toxin amounts detected were significant, above the Dutch limit value established in 2017, showing the importance and the need for continuity of these studies to gain more knowledge about the prevalence of these toxins, unraveling new vectors, in order to better assess human health risk. This work represents a general overview of new TTX bearers (7) most of them in gastropods (Patella depressa, Nucella lapillus, Onchidella celtica and Aplysia depilans), followed by echinoderms (Echinus esculentus and Ophidiaster ophidianus) and puffer fish Sphoeroides marmoratus.

Paralytic Shellfish Toxins Occurrence in Non-Traditional Invertebrate Vectors from North Atlantic Waters (Azores, Madeira, and Morocco).

Paralytic shellfish toxins (PSTs) are potent alkaloids of microalgal and cyanobacterial origin, with worldwide distribution. Over the last 20 years, the number of poisoning incidents has declined as a result of the implementation of legislation and monitoring programs based on bivalves. In the summer of 2012 and 2013, we collected a total of 98 samples from 23 different species belonging to benthic and subtidal organisms, such as echinoderms, crustaceans, bivalves, and gastropods. The sampling locations were Madeira, São Miguel Island (Azores archipelago), and the northwestern coast of Morocco. The samples were analyzed using post-column oxidation liquid chromatography with a fluorescence detection method. Our main goal was to detect new vectors for these biotoxins. After reporting a total of 59 positive results for PSTs with 14 new vectors identified, we verified that some of the amounts exceeded the limit value established in the EU. These results suggest that routine monitoring of saxitoxin and its analogs should be extended to more potential vectors other than bivalves, including other edible organisms, for a better protection of public health.

Ocean currents and acoustic backscatter data from shipboard ADCP measurements at three North Atlantic seamounts between 2004 and 2015.

Seamounts are amongst the most common physiographic structures of the deep-ocean landscape, but remoteness and geographic complexity have limited the systematic collection of integrated and multidisciplinary data in the past. Consequently, important aspects of seamount ecology and dynamics remain poorly studied. We present a data collection of ocean currents and raw acoustic backscatter from shipboard Acoustic Doppler Current Profiler (ADCP) measurements during six cruises between 2004 and 2015 in the tropical and subtropical Northeast Atlantic to narrow this gap. Measurements were conducted at seamount locations between the island of Madeira and the Portuguese mainland (Ampère, Seine Seamount), as well as east of the Cape Verde archipelago (Senghor Seamount). The dataset includes two-minute ensemble averaged continuous velocity and backscatter profiles, supplemented by spatially gridded maps for each velocity component, error velocity and local bathymetry. The dataset is freely available from the digital data library PANGAEA at https://doi.pangaea.de/10.1594/PANGAEA.883193.

New Invertebrate Vectors of Okadaic Acid from the North Atlantic Waters--Portugal (Azores and Madeira) and Morocco.

Okadaic acid and its analogues are potent phosphatase inhibitors that cause Diarrheic Shellfish Poisoning (DSP) through the ingestion of contaminated shellfish by humans. This group of toxins is transmitted worldwide but the number of poisoning incidents has declined over the last 20 years due to legislation and monitoring programs that were implemented for bivalves. In the summer of 2012 and 2013, we collected a total of 101 samples of 22 different species that were made up of benthic and subtidal organisms such echinoderms, crustaceans, bivalves and gastropods from Madeira, São Miguel Island (Azores archipelago) and the northwestern coast of Morocco. The samples were analyzed by UPLC-MS/MS. Our main objective was to detect new vectors for these biotoxins. We can report nine new vectors for these toxins in the North Atlantic: Astropecten aranciacus, Arbacia lixula, Echinaster sepositus, Holothuria sanctori, Ophidiaster ophidianus, Onchidella celtica, Aplysia depilans, Patella spp., and Stramonita haemostoma. Differences in toxin contents among the species were found. Even though low concentrations were detected, the levels of toxins that were present, especially in edible species, indicate the importance of these types of studies. Routine monitoring should be extended to comprise a wider number of vectors other than for bivalves of okadaic acid and its analogues.

First Report of Ciguatoxins in Two Starfish Species: Ophidiaster ophidianus and Marthasterias glacialis.

Ciguatera fish poisoning (CFP) is a syndrome caused by the ingestion of fish contaminated with Ciguatoxins (CTXs). These phycotoxins are produced mainly by dinoflagellates that belong to the genus Gambierdiscus that are transformed in more toxic forms in predatory fish guts, and are more present in the Indo-Pacific and Caribbean areas. It is estimated that CFP causes per year more than 10,000 intoxications worldwide. With the rise of water temperature and anthropogenic intervention, it is important to study the prevalence of CFP in more temperate waters. Through inter- and subtidal sampling, 22 species of organisms were collected, in Madeira and Azores archipelagos and in the northwestern Moroccan coast, during September of 2012 and June and July of 2013. A total of 94 samples of 22 different species of bivalves, gastropods, echinoderms and crustaceans where analyzed by Ultra Performance Liquid Chromatography-Mass Spectometry-Ion Trap-Time of Flight (UPLC-MS-IT-TOF) and Ultra Performance Chromatography- Mass Spectrometry (UPLC-MS). Our main aim was to detect new vectors and ascertain if there were some geographical differences. We detected for the first time putative CTXs in echinoderms, in two starfish species-M. glacialis and O. ophidianus. We detected differences regarding uptake values by organisms and geographical location. Toxin amounts were significant, showing the importance and the need for continuity of these studies to gain more knowledge about the prevalence of these toxins, in order to better access human health risk. In addition, we suggest monitoring of these toxins should be extended to other vectors, starfish being a good alternative for protecting and accessing human health risk.

New species and new records of bryozoans from shallow waters of Madeira Island.

Two new species of bryozoans encrusting subtidal rocks are described from the shallow waters of Madeira Island. We describe one cyclostome, Favosipora purpurea sp. nov., which represents the first record of this genus in the Atlantic Ocean, and one cheilostome, Rhynchozoon papuliferum sp. nov. In addition, one species, Beania maxilladentata, is recorded for the first time outside of Rio de Janeiro, Brazil. Six other species previously recorded in Madeira are redescribed to provide new data and SEM images.

Long-term tumor remission under trastuzumab treatment for HER2 positive metastatic breast cancer - results from the HER-OS patient registry.

BACKGROUND: In this study, we examined patients who had non-progressive disease for at least 2 years after diagnosis of inoperable locoregional recurrent or metastatic breast cancer under continuous trastuzumab treatment. Our primary goal was to assess the long-term outcome of patients with durable response to trastuzumab. METHODS: 268 patients with HER2-positive inoperable locally recurrent or metastatic breast cancer and non-progressive disease for at least 2 years under trastuzumab treatment were documented retrospectively or prospectively in the HER-OS registry, an online documentation tool, between December 2006 and September 2010 by 71 German oncology centers. The study end point was time to tumor progression. RESULTS: Overall, 47.1% of patients (95% confidence interval (CI): 39.9-54.1%) remained in remission for more than 5 years, while the median time to progression was 4.5 years (95% CI: 4.0-6.6 years). Lower age (<50 years) and good performance status (ECOG 0) at time of trastuzumab treatment initiation as well as complete remission after initial trastuzumab treatment were associated with longer time to progression. Interruption of trastuzumab therapy correlated with shorter time to progression. CONCLUSIONS: HER2-positive patients, who initially respond to palliative treatment with trastuzumab, can achieve a long-term tumor remission of several years.

Breakthroughs in research and treatment of early breast cancer: an overview of the last three decades.

Breast cancer has become curable for the majority of women in Western Europe and North America. Advances have been made in imaging diagnostics as well as the implementation of nationwide screening programmes. Nowadays, we talk about prevention as well as treatment. Pathology has moved from pure morphology (tumour type, grade and stage) to biological characterisation of the tumour. Treatment has changed considerably through a better understanding of the disease; from a local disease predominated by extensive and mutilating surgical techniques to a point where breast cancer has come into its own as a systemic disease with equal "rights" to local as well as systemic treatment. This paradigm shift has led to a multidisciplinary approach of the understanding and treatment of breast cancer. Molecular classification has changed the understanding of breast cancer and will be the basis for an even more individualised treatment. New (biological) agents will help to further tailor treatment to response or resistance. While systemic treatment has been increased in number and duration surgical/local strategies have been reduced to minimum. Evidence-based medicine has helped to improve and standardise treatment of breast cancer. This review summarises the 10th Biedenkopf meeting that was held to review the advances in breast cancer understanding and treatment.

Response-guided neoadjuvant chemotherapy for breast cancer.

PURPOSE: We investigated disease-free survival (DFS) and overall survival (OS) after response-guided neoadjuvant chemotherapy in patients with early breast cancer. PATIENTS AND METHODS: We treated 2,072 patients with two cycles of docetaxel, doxorubicin, and cyclophosphamide (TAC) and randomly assigned early responders to four (n = 704) or six (n = 686) additional TAC cycles, and early nonresponders to four cycles of TAC (n = 321) or vinorelbine and capecitabine (NX; n = 301) before surgery. RESULTS: DFS was longer in early responders receiving TAC × 8 than in those receiving TAC × 6 (hazard ratio [HR], 0.78; 95% CI, 0.62 to 0.97; P = .026), and in early nonresponders receiving TAC-NX than in those receiving TAC × 6 (HR, 0.59; 95% CI, 0.49 to 0.82; P = .001). Exploratory analysis showed that DFS after response-guided chemotherapy (TAC × 8 or TAC-NX) was significantly longer (HR, 0.71; 95% CI, 0.60 to 0.85; P < .003), as was OS (HR, 0.79; 95% CI, 0.63 to 0.99; P = .048), than on conventional chemotherapy (TAC × 6). DFS was longer after response-guided chemotherapy in all hormone receptor-positive tumors (luminal A HR = 0.55, luminal B [human epidermal growth factor receptor 2 (HER2) negative] HR = 0.40, and luminal B [HER2 positive] HR = 0.56), but not in hormone receptor-negative tumors (HER2 positive [nonluminal] HR = 1.01 and triple negative HR = 0.87). Pathologic complete response did not predict these survival effects. pCR predicted an improved DFS in triple-negative (HR = 6.67), HER2-positive (nonluminal; HR 5.24), or luminal B (HER2-negative) tumors (HR = 3.74). CONCLUSION: This exploratory analysis suggests that response-guided neoadjuvant chemotherapy might improve survival and is most effective in hormone receptor-positive tumors. If confirmed, the response-guided approach could provide a clinically meaningful advantage for the neoadjuvant over the adjuvant approach in early breast cancer.

Adjuvant lapatinib for women with early-stage HER2-positive breast cancer: a randomised, controlled, phase 3 trial.

BACKGROUND: Worldwide, many patients with HER2-positive early stage breast cancer do not receive trastuzumab-the standard adjuvant treatment. We investigated the efficacy and safety of adjuvant lapatinib for patients with trastuzumab-naive HER2-positive early-stage breast cancer, started at any time after diagnosis. METHODS: This study was a placebo-controlled, multicentre, randomised phase 3 trial. Women outpatients from 405 [corrected] centres in 33 countries [corrected] with HER2-positive early-breast cancer who had previously received adjuvant chemotherapy but not trastuzumab were randomly assigned (1:1) to receive daily lapatinib (1500 mg) or daily placebo for 12 months. Randomisation was done with a computer-generated sequence, stratified by time since diagnosis, lymph node involvement at diagnosis, and tumour hormone-receptor status. Investigators, site staff, and patients were masked to treatment assignment. The primary endpoint was disease-free survival in the intention-to-treat population. This study is registered with ClinicalTrials.gov, number NCT00374322. FINDINGS: Between August, 2006, and May, 2008, 3161 women were enrolled and 3147 were assigned to lapatinib (n=1571) or placebo (n=1576). After a median follow-up of 47·4 months (range 0·4-60·0) in the lapatinib group and 48·3 (0·7-61·3) in the placebo group, 210 (13%) disease-free survival events had occurred in the lapatinib group versus 264 (17%) in the placebo group (hazard ratio [HR] 0·83, 95% CI 0·70-1·00; p=0·053). Central review of HER2 status showed that only 2490 (79%) of the randomised women were HER2-positive. 157 (13%) of 1230 confirmed HER2-positive patients in the lapatinib group and in 208 (17%) of 1260 in the placebo group had a disease-free survival event (HR 0·82, 95% 0·67-1·00; p=0·04). Serious adverse events occurred in 99 (6%) of 1573 patients taking lapatinib and 77 (5%) of 1574 patients taking placebo, with higher incidences of grade 3-4 diarrhoea (97 [6%] vs nine [<1%]), rash (72 [5%] vs three [<1%]), and hepatobiliary disorders (36 [2%] vs one [<1%]). INTERPRETATION: Our data show that there was no significant difference in disease-free survival between groups when analysed in the intention-to-treat population. However, exploratory analyses restricted to patients who had HER2-positive disease confirmed by central fluorescence in-situ hybridisation review suggested marginal benefit with lapatinib in terms of disease-free survival. Thus lapatinib might be an option for women with HER2-positive breast cancer who do not or cannot receive adjuvant trastuzumab. FUNDING: GlaxoSmithKline.