ESMO Expert Consensus Statements on Cancer Survivorship: promoting high-quality survivorship care and research in Europe.

BACKGROUND: The increased number of cancer survivors and the recognition of physical and psychosocial challenges, present from cancer diagnosis through active treatment and beyond, led to the discipline of cancer survivorship. DESIGN AND METHODS: Herein, we reflected on the different components of survivorship care, existing models and priorities, in order to facilitate the promotion of high-quality European survivorship care and research. RESULTS: We identified five main components of survivorship care: (i) physical effects of cancer and chronic medical conditions; (ii) psychological effects of cancer; (iii) social, work and financial effects of cancer; (iv) surveillance for recurrences and second cancers; and (v) cancer prevention and overall health and well-being promotion. Survivorship care can be delivered by structured care models including but not limited to shared models integrating primary care and oncology services. The choice of the care model to be implemented has to be adapted to local realities. High-quality care should be expedited by the generation of: (i) focused and shared European recommendations, (ii) creation of tools to facilitate implementation of coordinated care and (iii) survivorship educational programs for health care teams and patients. The research agenda should be defined with the participation of health care providers, researchers, policy makers, patients and caregivers. The following patient-centered survivorship research areas were highlighted: (i) generation of a big data platform to collect long-term real-world data in survivors and healthy controls to (a) understand the resources, needs and preferences of patients with cancer, and (b) understand biological determinants of survivorship issues, and (ii) develop innovative effective interventions focused on the main components of survivorship care. CONCLUSIONS: The European Society for Medical Oncology (ESMO) can actively contribute in the efforts of the oncology community toward (a) promoting the development of high-quality survivorship care programs, (b) providing educational material and (c) aiding groundbreaking research by reflecting on priorities and by supporting research networking.

Supporting the gastrointestinal microenvironment during high-dose chemotherapy and stem cell transplantation by inhibiting IL-1 signaling with anakinra.

High-dose chemotherapy causes intestinal inflammation and subsequent breakdown of the mucosal barrier, permitting translocation of enteric pathogens, clinically manifesting as fever. Antibiotics are mainstay for controlling these complications, however, they are increasingly recognized for their detrimental effects, including antimicrobial resistance and dysbiosis. Here, we show that mucosal barrier injury induced by the mucotoxic chemotherapeutic agent, high-dose melphalan (HDM), is characterized by hyper-active IL-1b/CXCL1/neutrophil signaling. Inhibition of this pathway with IL-1RA, anakinra, minimized the duration and intensity of mucosal barrier injury and accompanying clinical symptoms, including diarrhea, weight loss and fever in rats. 16S analysis of fecal microbiome demonstrated a more stable composition in rats receiving anakinra, with reduced pathogen expansion. In parallel, we report through Phase IIA investigation that anakinra is safe in stem cell transplant patients with multiple myeloma after HDM. Ramping-up anakinra (100-300 mg administered intravenously for 15 days) did not cause any adverse events or dose limiting toxicities, nor did it change time to neutrophil recovery. Our results reinforce that strengthening the mucosal barrier may be an effective supportive care strategy to mitigate local and systemic clinical consequences of HDM. We are now conducting a Phase IIB multicenter, placebo-controlled, double-blinded trial to assess clinical efficacy of anakinra (AFFECT-2).Trial registration: ClinicalTrials.gov identifier: NCT03233776.

Development of a self-limiting model of methotrexate-induced mucositis reinforces butyrate as a potential therapy.

Gastrointestinal mucositis is a complication of anticancer treatment, with few validated in vitro systems suitable to study the complex mechanisms of mucosal injury. Therefore, we aimed to develop and characterize a chemotherapeutic-induced model of mucositis using 3D intestinal organoids. Organoids derived from mouse ileum were grown for 7 days and incubated with different concentrations of the chemotherapeutic agent methotrexate (MTX). Metabolic activity, citrulline levels and cytokine/chemokine production were measured to determine the optimal dosage and incubation time. The protective effects of folinic acid on the toxicity of MTX were investigated by pre-treating organoids with (0.0005-50 µg/mL) folinic acid. The impact of microbial-derived short-chain fatty acids was evaluated by supplementation with butyrate in the organoid model. MTX caused a dose-dependent reduction in cell metabolic activity and citrulline production that was salvaged by folinic acid treatment. Overall, MTX causes significant organoid damage, which can be reversed upon removal of MTX. The protective effect of folinic acid suggest that the organoids respond in a clinical relevant manner. By using the model for intervention, it was found that prophylactic treatment with butyrate might be a valuable strategy for prophylactic mucositis prevention.

Safety of assisted reproductive techniques in young women harboring germline pathogenic variants in BRCA1/2 with a pregnancy after prior history of breast cancer.

BACKGROUND: Knowledge is growing on the safety of assisted reproductive techniques (ART) in cancer survivors. No data exist, however, for the specific population of breast cancer patients harboring germline BRCA1/2 pathogenic variants. PATIENTS AND METHODS: This is a multicenter retrospective cohort study across 30 centers worldwide including women diagnosed at ≤40 years with stage I-III breast cancer, between January 2000 and December 2012, harboring known germline BRCA1/2 pathogenic variants. Patients included in this analysis had a post-treatment pregnancy either achieved through use of ART (ART group) or naturally (non-ART group). ART procedures included ovulation induction, ovarian stimulation for in vitro fertilization or intracytoplasmic sperm injection, and embryo transfer under hormonal replacement therapy. RESULTS: Among the 1424 patients registered in the study, 168 were eligible for inclusion in the present analysis, of whom 22 were in the ART group and 146 in the non-ART group. Survivors in the ART group conceived at an older age compared with those in the non-ART group (median age: 39.7 versus 35.4 years, respectively). Women in the ART group experienced more delivery complications compared with those in the non-ART group (22.1% versus 4.1%, respectively). No other apparent differences in obstetrical outcomes were observed between cohorts. The median follow-up from pregnancy was 3.4 years (range: 0.8-8.6 years) in the ART group and 5.0 years (range: 0.8-17.6 years) in the non-ART group. Two patients (9.1%) in the ART group experienced a disease-free survival event (specifically, a locoregional recurrence) compared with 40 patients (27.4%) in the non-ART group. In the ART group, no patients deceased compared with 10 patients (6.9%) in the non-ART group. CONCLUSION: This study provides encouraging safety data on the use of ART in breast cancer survivors harboring germline pathogenic variants in BRCA1/2, when natural conception fails or when they opt for ART in order to carry out preimplantation genetic testing.

Translational model of melphalan-induced gut toxicity reveals drug-host-microbe interactions that drive tissue injury and fever.

PURPOSE: Conditioning therapy with high-dose melphalan (HDM) is associated with a high risk of gut toxicity, fever and infections in haematopoietic stem cell transplant (HSCT) recipients. However, validated preclinical models that adequately reflect clinical features of melphalan-induced toxicity are not available. We therefore aimed to develop a novel preclinical model of melphalan-induced toxicity that reflected well-defined clinical dynamics, as well as to identify targetable mechanisms that drive intestinal injury. METHODS: Male Wistar rats were treated with 4-8 mg/kg melphalan intravenously. The primary endpoint was plasma citrulline. Secondary endpoints included survival, weight loss, diarrhea, food/water intake, histopathology, body temperature, microbiota composition (16S sequencing) and bacterial translocation. RESULTS: Melphalan 5 mg/kg caused self-limiting intestinal injury, severe neutropenia and fever while impairing the microbial metabolome, prompting expansion of enteric pathogens. Intestinal inflammation was characterized by infiltration of polymorphic nuclear cells in the acute phases of mucosal injury, driving derangement of intestinal architecture. Ileal atrophy prevented bile acid reabsorption, exacerbating colonic injury via microbiota-dependent mechanisms. CONCLUSION: We developed a novel translational model of melphalan-induced toxicity, which has excellent homology with the well-known clinical features of HDM transplantation. Application of this model will accelerate fundamental and translational study of melphalan-induced toxicity, with the clinical parallels of this model ensuring a greater likelihood of clinical success.

Pre-therapy fasting slows epithelial turnover and modulates the microbiota but fails to mitigate methotrexate-induced gastrointestinal mucositis.

BACKGROUND: Recent findings by Tang et al. (2020) show dietary restriction (30%, 2 weeks) prevents methotrexate-induced mortality by modulation of the microbiota, specifically the expansion of Lactobacillus. While fundamentally insightful, upscaling this schedule is a major obstacle to clinical uptake. Here, we evaluate a safe and clinically achievable schedule of pre-therapy fasting for 48 h on microbiota composition, body composition and intestinal proliferation, and assess its impact on the severity of methotrexate-induced gastrointestinal mucositis using a validated preclinical rat model. METHODS: Age- and weight-matched male Wistar rats were treated with a sublethal dose of 45 mg/kg methotrexate with or without pre-therapy fasting. The impact of acute fasting on epithelial proliferation, body composition and the microbiota was assessed using plasma citrulline, Ki67 immunohistochemistry, miniSpec and 16S rRNA sequencing. The severity of gastrointestinal mucositis was evaluated using plasma citrulline and body weight. RESULTS: Whilst pre-therapy fasting slowed epithelial proliferation and increased microbial diversity and richness, it also induced significant weight loss and was unable to attenuate the severity of mucositis in both age- and weight-matched groups. In contrast to Tang et al., we saw no expansion of Lactobacillus following acute fasting. CONCLUSIONS: Our findings suggest that the beneficial effects of acute fasting are masked by the detrimental effects on body weight and composition and lacking influence on Lactobacillus. Future studies should consider alternative fasting schedules or aim to induce comparable microbial and mucosal manipulation without compromising body composition using clinically feasible methods of dietary or microbial intervention.

Differential impact of endocrine therapy and chemotherapy on quality of life of breast cancer survivors: a prospective patient-reported outcomes analysis.

BACKGROUND: In early breast cancer (BC), there has been a trend to escalate endocrine therapy (ET) and to de-escalate chemotherapy (CT). However, the impact of ET versus CT on the quality of life (QoL) of early BC patients is unknown. Here, we characterize the independent contribution of ET and CT on patient-reported outcomes (PROs) at 2 years after diagnosis. PATIENTS AND METHODS: We prospectively collected PROs in 4262 eligible patients using the European Organization for Research and Treatment of Cancer QLQ-C30/BR23 questionnaires inside CANTO trial (NCT01993498). The primary outcome was the C30 summary score (C30-SumSc) at 2 years after diagnosis. RESULTS: From eligible patients, 37.2% were premenopausal and 62.8% postmenopausal; 81.9% received ET and 52.8% CT. In the overall cohort, QoL worsened by 2 years after diagnosis in multiple functions and symptoms; exceptions included emotional function and future perspective, which improved over time. ET (Pint = 0.004), but not CT (Pint = 0.924), had a persistent negative impact on the C30-SumSc. In addition, ET negatively impacted role and social function, pain, insomnia, systemic therapy side-effects, breast symptoms and further limited emotional function and future perspective recovery. Although CT had no impact on the C30-SumSc at 2-years it was associated with deteriorated physical and cognitive function, dyspnea, financial difficulties, body image and breast symptoms. We found a differential effect of treatment by menopausal status; in premenopausal patients, CT, despite only a non-significant trend for deteriorated C30-SumSc (Pint = 0.100), was more frequently associated with QoL domains deterioration than ET, whereas in postmenopausal patients, ET was more frequently associated with QoL deterioration, namely using the C30-SumSc (Pint = 0.004). CONCLUSION(S): QoL deterioration persisted at 2 years after diagnosis with different trajectories by treatment received. ET, but not CT, had a major detrimental impact on C30-SumSc, especially in postmenopausal women. These findings highlight the need to properly select patients for adjuvant ET escalation.

Reproductive potential and performance of fertility preservation strategies in BRCA-mutated breast cancer patients.

Background: Preclinical evidence suggests a possible negative impact of deleterious BRCA mutations on female fertility. However, limited and rather conflicting clinical data are available. This study assessed the reproductive potential and performance of fertility preservation strategies in BRCA-mutated breast cancer patients. Patients and methods: This was a retrospective analysis of two prospective studies investigating oocyte cryopreservation and ovarian tissue cryopreservation in newly diagnosed early breast cancer patients. In the current analysis, baseline anti-Mullerian hormone (AMH) and performance of cryopreservation strategies were compared between patients with or without germline deleterious BRCA mutations. Results: Out of 156 patients included, 101 had known BRCA status of whom 29 (18.6%) were BRCA-mutated and 72 (46.1%) had no mutation. Median age in the entire cohort was 31 years [interquartile range (IQR) 28-33). Median AMH levels were 1.8 µg/l (IQR 1.0-2.7) and 2.6 µg/l (IQR 1.5-4.1) in the BRCA-positive and BRCA-negative cohorts, respectively (P = 0.109). Among patients who underwent oocyte cryopreservation (N = 29), women in the BRCA-positive cohort tended to retrieve (6.5 versus 9; P = 0.145) and to cryopreserve (3.5 versus 6; P = 0.121) less oocytes than those in the BRCA-negative cohort. Poor response rate (i.e. retrieval of ≤4 oocytes) was 40.0% and 11.1% in the BRCA-positive and BRCA-negative cohorts, respectively (P = 0.147). Among patients who underwent ovarian tissue cryopreservation (N = 72), women in the BRCA-positive cohort tended to have a numerically lower number of oocytes per fragment (0.08 versus 0.14; P = 0.193) and per square millimeter (0.33 versus 0.78; P = 0.153) than those in the BRCA-negative cohort. Two BRCA-mutated patients were transplanted after chemotherapy and one delivered at term a healthy baby. No difference between BRCA1- and BRCA2-mutated patients was observed in any of the above-mentioned outcomes. Conclusion: A consistent trend for reduced reproductive potential and performance of cryopreservation strategies was observed in BRCA-mutated breast cancer patients. Independent validation of these results is needed.

Serum YB-1 (Y-box binding protein 1) as a biomarker of bone disease progression in patients with breast cancer and bone metastases.

YB-1 (Y-box binding protein 1) is a multifunctional cold-shock protein that has been implicated in all hallmarks of cancer. Elevated YB-1 protein level was associated with poor prognosis in several types of cancers, including breast cancer (BC), where it is a marker of decreased overall survival (OS) and distant metastasis-free survival across all subtypes. YB-1 is also secreted by different cell types and may act as an extracellular mitogen; however the pathological implications of the secreted form of YB-1 (sYB-1) are unknown. Our purpose was to retrospectively evaluate the association between YB-1 measured by ELISA in serum and disease characteristics and outcomes in patients with BC and bone metastases (BM). In our cohort, sYB-1 was detected in the serum of 22 (50%) patients, and was associated with the presence of extra-bone metastases (p=0.044). Positive sYB-1 was also associated with faster bone disease progression (HR 3.1, 95% CI 1.09-8.95, P=0.033), but no significant differences were observed concerning OS, and time to development of skeletal-related events. Moreover, patients with positive sYB-1 also had higher levels of IL-6, a known osteoclastogenic inducer. Therefore, detection of sYB-1 in patients with BC and BM may indicate a higher tumor burden, in bone and extra-bone locations, and is a biomarker of faster bone disease progression.

Comparação entre as abordagens lateral direita e mediana ventral em cadelas submetidas à ovariossalpingo-histerectomia / Comparison between the right lateral and ventral median approach in a bitch undergoing ovariosalpinghysterectomy

A fim de comparar as abordagens abdominais, pela linha mediana ventral e lateral direita em cadelas pré-púberes e adultas submetidas a ovariossalpingo-histerectomia, utilizaram-se 28 cadelas hígidas, distribuídas em dois grupos experimentais de igual número: grupo abordagem mediana ventral (AMV) e grupo abordagem lateral direita (ALD), com sete animais adultos e sete animais pré-púberes em cada grupo. O procedimento cirúrgico foi dividido em nove manobras cirúrgicas distintas, e o tempo para conclusão de cada uma delas, suas facilidades e dificuldades, assim como o tempo cirúrgico total, foram determinados. O tempo médio desde o início da incisão da pele até a entrada na cavidade peritoneal foi menor nas cadelas adultas (P≤0,001) e pré-púberes (P≤0,001) do grupo AMV, mas o tempo médio para identificação uterina foi menor nas cadelas pré-púberes (P≤0,001) do grupo ALD. O tempo cirúrgico total foi menor utilizando-se a abordagem lateral direita (grupo ALD) nas cadelas adultas (P≤0,001) e pré-púberes (P≤0,001). Seu uso não se relacionou com complicações cirúrgicas e facilitou a identificação uterina, possibilitando redução no tempo cirúrgico total. Assim, a abordagem lateral direita demonstrou ser uma alternativa segura em cadelas adultas e pré-púberes submetidas à OSH eletiva.

In order to compare the abdominal approaches, through the ventral midline and right lateral in pre-pubertal adult female dogs undergoing ovariohysterectory we used 28 otherwise healthy dogs, divided into two experimental groups of equal number: Ventral Median Approach Group (VMA) and Right Side Approach Group (RSA), with seven adult animals and seven pre-pubertal animals in each group. The surgical procedure was divided into nine different surgical maneuvers, and the time required for completion of each of them, their strengths and difficulties, as well as the total surgical time were determined. The time from the start of the skin incision to the entrance into the peritoneal cavity was lower in adult female dogs (P≤0.001) and pre-pubertal (P≤0.001) in the VAM group, but the time for uterine identification was lower in pre-pubertal female dogs (P≤ 0.001) in the RSA group. The total surgical time was shorter using the right lateral approach (RSA group) in adult (P≤ 0.001) and pre-pubertal (P≤0.001) female dogs. Thus, the right-side approach has proved to be a safe alternative in adult and pre-pubertal dogs undergoing elective OSH. Its use was not associated with surgical complications, and facilitated uterine identification, allowing a reduction in the total surgical time.

O uso do peritônio de paca conservado em solução supersaturada de açúcar a 300 por cento ou glicerina a 98 por cento implantados na parede abdominal de ratos / The use of paca peritoneum preserved in supersaturated sugar solution 300 percent or glycerine 98 percent implanted in the abdominal wall of rats

Na busca de material biológico alternativo para a realização de implantes, objetivou-se com o presente estudo avaliar comparativamente a implantação do peritônio de paca, uma nova opção de biomaterial, conservado em solução supersaturada de açúcar a 300 por cento e conservado em glicerina a 98 por cento na parede abdominal de ratos Wistar. Foram utilizados 60 ratos, machos, da linhagem Wistar, pesando entre 150 e 200 gramas, organizados nos seguintes grupos experimentais: grupo controle (GI), grupo peritônio conservado em solução supersaturada de açúcar a 300 por cento (GII) e grupo peritônio conservado em glicerina a 98 por cento (GIII), cada um com 20 animais. Os grupos GII e GIII receberam o enxerto de peritônio da paca conservado em solução de açúcar 300 por cento e glicerina 98 por cento, respectivamente, e o grupo GI não recebeu a membrana. Cinco ratos de cada grupo foram submetidos à eutanásia em quatro momentos distintos: sete, 15, 30 e 60 dias de pós-operatório para avaliações macroscópicas e microscópicas da interface implante-tecido nativo. Apesar de reações adversas observadas em 57,5 por cento dos animais do grupo GII e GIII, em 95 por cento dos animais desses grupos houve boa cicatrização da membrana. Na análise histológica, verificou-se a presença de grande infiltrado inflamatório nos períodos iniciais (sete e 15 dias) e grande presença de tecido conjuntivo nos momentos finais (30 e 60 dias). Concluiu-se que o peritônio da paca como membrana biológica conservado nos meios estudados pode ser utilizado com segurança na parede abdominal de ratos; ainda, que sua conservação em solução supersaturada de açúcar a 300 por cento permitiu melhor maleabilidade no ato cirúrgico...

In the search for alternative biological material to perform implants, this study aimed to compare the implantation of paca peritoneum, a new biomaterial option, preserved in supersaturated sugar solution 300 percent and preserved in glycerin 98 percent in the abdominal wall of Wistar rats. A total of 60 male rats from the Wistar strain weighing between 150 and 200 grams were housed into three different experimental groups: control group (GI), peritoneum preserved in supersaturated sugar solution 300 percent group (GII) and peritoneum preserved in glycerin 98 percent group (GIII), with 20 animals each. The GII and GIII received the paca peritoneum graft preserved in sugar solution 300 percent and glycerin 98 percent, respectively and group GI did not receive any membrane. Five rats from each group were euthanized at four different times: seven, 15, 30 and 60 days post-surgery for macroscopic and microscopic evaluations in graft-native tissue interface. Despite the adverse reactions observed in 57.5 percent of GII and GIII, there was good healing of the membrane in 95 percent of the animals in these groups. Upon histological examination there was a large presence of inflammatory infiltrates in the initial periods (seven and 15 days) and a large presence of connective tissue in the final stages (30 and 60). It was concluded that the paca peritoneum as biological membrane preserved as presented in this study can be used safely in the abdominal wall of rats, the preservation in supersaturated sugar solution 300 percent allowed better flexibility during surgery...

Avaliação macroscópica e microscópica de implante de Biomembrana em estômago de coelhos (Oryctolagus cuniculus) / Macroscopic and microscopic evaluation biomembrane implant in the stomach of rabbits (Oryctolagus cuniculus)

Neste estudo foi implantado um retalho de biomembrana de látex natural em substituição a um fragmento da parede de estômago de coelhos albinos, raça Nova Zelândia, adultos, machos não castrados (n=12), com o intuito de avaliar o processo de reparação tecidual no que se refere à biocompatibilidade, à capacidade de reparação tecidual e a possíveis complicações. Aos 15, 30 e 60 dias de pós-operatório, os animais foram sacrificados, mediante o emprego de tiopental sódico (200mg kg-1), para posterior avaliação macroscópica e análise histopatológica da interface do implante com o tecido nativo. Macroscopicamente, aos 15, 30 e 60 dias de pós-operatório observou-se presença de aderências na face serosa. Aos 60 dias de pós-operatório, em todos os animais avaliados, a biomembrana não foi encontrada. Sob microscopia de luz, aos 15 e 30 dias, verificaram-se descontinuidade das camadas muscular e mucosa, presença de infiltrado inflamatório polimorfonuclear. Foram visualizados vasos sanguíneos e fibras musculares. Aos 60 dias, as camadas mucosa, muscular estavam completamente reconstituídas. O implante foi biocompatível e forneceu arcabouço para orientação e desenvolvimento das camadas teciduais do estômago, mediante processos de reparação, restabelecendo a estrutura do órgão.

In this study, biomembrane of natural latex was utilized to replace a section of the stomach wall of New Zealand rabbits, adult and non-castrated males (n=12), in order to evaluate the tissue repair process in regards to its biocompatility, scar formation ability and possible complications. The animals were euthanized at fifteen, 30 and 60 days post operation, by use of sodium thiopental (200mg kg-1), followed by macroscopic and histopathological analysis of the implant interface with the native tissue. Macroscopically, at fifteen, 30, and 60 days post operation adherence was observed in the serosal wall. At 60 days post operation, the biomembrane is not in the stomach. Under light microscopy, at fifteen and 30 days, discontinuity of muscle layer and mucosa layer, and presence of polimorfonuclear population of inflammatory cells was observed. New vessels and muscle fibers were observed. At 60 days, the mucosa and muscle layers were complete reconstituted. The implants were biocompatible and had provided the mainframe for orientation and development of the tissue layers through repairing processes, thus reestablishing the organ structure.

Estudo retrospectivo de ovariossalpingo-histerectomia em cadelas e gatas atendidas em Hospital Veterinário Escola no período de um ano / Retrospective ovariosalpingohisterectomy study in bitches and queens assisted at a Veterinary School Hospital during one year

Realizou-se um estudo retrospectivo das indicações de ovariossalpingo-histerectomia - eletiva e terapêutica - no período de um ano. Foram analisados 193 prontuários de cadelas e gatas atendidas em Hospital Veterinário Escola, entre março de 2010 e março de 2011, levando em consideração a espécie, a idade e o uso ou não de anticoncepcional. Constatou-se que a demanda de OSH com caráter terapêutico (78,8%) é consideravelmente mais alta que a eletiva (21,2%). Observou-se que as anormalidades reprodutivas detectadas com maior frequência foram piometra (53,36%) e complicações obstétricas (25,38%) e que o uso de anticoncepcional foi o fator relevante para o delineamento desses quadros.

An ovariosalpingohisterectomy procedure indication - elective and therapeutic - retrospective study was conducted during one year. Record files from 193 bitches and queens assisted at a Veterinary School Hospital between March 2010 and March 2011 were analyzed, considering species, age and contraceptive use. It was possible to conclude that the therapeutic OSH demand (78.8%) is remarkably superior to the elective OSH request (21.2%). It was also observed that the most frequently detected reproductive abnormalities were pyometra (53.36%) and gestational complications (25.38%), and also that the use of contraceptives is a relevant factor for those events.

Determination of bisphenol A and bisphenol B in canned seafood combining QuEChERS extraction with dispersive liquid-liquid microextraction followed by gas chromatography-mass spectrometry.

A new simple and reliable method combining an acetonitrile partitioning extractive procedure followed by dispersive solid-phase cleanup (QuEChERS) with dispersive liquid-liquid microextraction (DLLME) and further gas chromatography mass spectrometry analysis was developed for the simultaneous determination of bisphenol A (BPA) and bisphenol B (BPB) in canned seafood samples. Besides the great enrichment factor provided, the final DLLME extractive step was designed in order to allow the simultaneous acetylation of the compounds required for their gas chromatographic analysis. Tetrachloroethylene was used as extractive solvent, while the acetonitrile extract obtained from QuEChERS was used as dispersive solvent, and anhydride acetic as derivatizing reagent. The main factors influencing QuEChERS and DLLME efficiency including nature of QuEChERS dispersive-SPE sorbents, amount of DLLME extractive and dispersive solvents and nature and amount of derivatizing reagent were evaluated. DLLME procedure provides an effective enrichment of the extract, allowing the required sensitivity even using a single quadropole MS as detector. The optimized method showed to be accurate (>68 % recovery), reproducible (<21 % relative standard deviation) and sensitive for the target analytes (method detection limits of 0.2 µg/kg for BPA and 0.4 µg/kg for BPB). The screening of several canned seafood samples commercialized in Portugal (total = 47) revealed the presence of BPA in more than 83 % of the samples with levels ranging from 1.0 to 99.9 µg/kg, while BPB was found in only one sample at a level of 21.8 µg/kg.

Plantas hipoglicemiantes utilizadas por comunidades tradicionais na Bacia do Alto Paraguai e Vale do Guaporé, Mato Grosso-Brasil

No Brasil, com inúmeras espécies tropicais, evidencia-se o conhecimento e uso da vegetação medicinal pelas comunidades quilombolas, ribeirinhas, rurais, tradicionais e indígenas, que são praticadas até os dias atuais. O Estado de Mato Grosso pela sua localização no Planalto Central brasileiro possui uma variedade destas comunidades, que utilizam estas plantas medicinais, levando em consideração o conhecimento popular passado de geração em geração. Esta pesquisa visa identificar a forma de coleta e os usos desse grupo vegetal indicados pelos usuários. O estudo foi realizado em 15 comunidades tradicionais (não indígenas) na Bacia do Alto Paraguai e 2 (duas) no Vale do Guaporé. A coleta de dados deu-se através de líderes comunitários, benzedeiras, parteiras e usuários utilizando o método qualitativo, mediante abordagem qualitativa, com auxílio de entrevistas estruturadas, semi-estruturadas e abertas. Dentre as espécies identificadas 17 estacam-se para o tratamento do Diabetes, dentre elas citam-se: Anacadium humile; Bauhinia glabra; Cecropia pachystachya; Hancornia speciosa; Heteropteris aphrodisiaca; Leonotis nepetifolia; Momordica charantia; Solanum lycocarpum. Registra-se ainda que as partes mais utilizadas desses vegetais são: folhas, casca do caule, raiz, planta toda, brotos, óleos dos frutos e polvilho dos frutos. De acordo com os usuários a maioria das plantas não podem ser colhidas após o nascer do sol e o preparo deve ser feito com folhas secas ou que tenham sido submetidas ao processo de secagem sobre o fogão de lenha ou em local abafado.

In Brazil, with a large number of tropical species, evidenciate the knowledge and use of the medicine vegetation by the "quilombolas", riverine, rurally, traditional and indigenous communities are used nowadays. The Mato Grosso state by their localization in the Central Brazilian Plateau posses a variety of community, which use those medicinal plants, carrying in consideration the popular knowledge passed though generation to generation. This research had as purpose to identificate the use of this vegetable group the collect ways related by the users and collectors. The study was realized into 15 traditional communities (not indigenous) in the Alto Paraguay Bay and 2 (two) in the Guaporé Valley. The data collect were made with the community leaders, women healers, midwife and users, it was used the qualitative method, with qualitative boarding, with aim of structured qualitative interview, semistructured and opened. Between the identified 17 destacated to the treatment of diabetes, between them it was cited Anacadium humile; Bauhinia glabra; Cecropia pachystachya; Hancornia speciosa; Heteropteris aphrodisiaca; Leonotis nepetifolia; Momordica charantia; Solanum lycocarpum. It was registered still, that the more used parts of this vegetable are: leaves, burke, root, all plant, shoot, fruit oil and fruits flour. In according to users the biggest part of the plants can't be collected after the sun rinsing and the prepare must be done with dry leaves or that had been submitted to the process of drying on the oven or in a muffled place.

Plantas medicinais usadas para tratamentos dermatológicos, em comunidades da Bacia do Alto Paraguai, Mato Grosso

O uso de plantas medicinais no Brasil e em especial no Estado de Mato Grosso vem sendo registrado em suas comunidades rurais e tradicionais. Este trabalho visa identificar plantas com potencial dermatológico utilizada pelas comunidades: quilombolas, ribeirinhas, rurais e tradicionais dos municípios de Barão de Melgaço, Nossa Senhora do Livramento, Poconé e Santo Antônio de Leverger. Fez-se a coleta de material botânico fértil e levantamento sobre o uso desses vegetais com a população local. Este material encontra-se identificado e depositado em Coleção de Referência de Plantas Medicinais no UFMT, Herbário Central. Registraram-se 36 espécies utilizadas no tratamento de doenças como afecções, feridas e úlceras, erisipelas, herpes labial e verrugas. Destacam-se espécies como: Astronium fraxinifolium Schott.; Bixa orellana L.; Brosimum gaudichaudii Tréc.; Cassia grandis Lf.; Heliotropium indicum L.; Jacaranda brasiliana (Lam.) Pers.; Macrosiphonia velame (St. Hil.) Mull. Arg.; Pistia stratiotes L.; Senna alatal. Roxb., entre outras. Estes vegetais são freqüentemente usados na forma de banhos e chás. O órgão vegetal mais utilizado foi as folhas, seguido de cascas do caule, frutos, óleo, raiz, planta inteira, seiva e sementes. O avanço da fronteira agropecuária nesse Estado é constante preocupação dos botânicos e das comunidades que ao longo das décadas usufruem desse potencial medicinal.

Medicinal plants used for dermatology treatment in communities of Alto Rio Paraguai' Basin in Mato Grosso State. The use of medicinal plants in Brazil and in special in the Mato Grosso State is being registered by the rural and traditional communities. This work aims to identify plants with dermatologic potential used by the communities: quilombolas, riversides, rural and traditional of the municipal districts of Barão de Melgaço, Nossa Senhora do Livramento, Poconé and Santo Antonio de Leverger. The collection of fertile botanical material and rising on the use of these vegetables was done with the local population. The identified material is deposited in the Reference Collection of Medicinal Plants in UFMT Central Herbarium. It were registered 36 species used in the diseases treatment as infections, hurt and ulcers, erysipelas, labial herpes and warts. Highlight species as: Astronium fraxinifolium Schott.; Bixa orellana L.; Brosimum gaudichaudii Tréc.; Cassia grandis Lf.; Heliotropium indicum L.; Jacaranda brasiliana (Lam.) Pers.; Macrosiphonia velame (St. Hil.) Mull. Arg.; Pistia stratiotes L.; Senna alata (l.) Roxb., come in another. These vegetables are frequently used in baths and teas form. The used vegetable organ was to the leaves followed by the peel and of the stem, fruits, oil, root, the whole plant, sap and seeds. The advance of the agriculture in this state is an constant preoccupation of the botanists and communities who along decades use those medicinal potential.

N-terminal chimeric constructs improve the expression of sarcoplasmic reticulum Ca(2+)-ATPase in yeast.

Wild-type and chimeric constructs comprising rabbit sarcoplasmic reticulum (SR) Ca(2+)-ATPase and the N-terminal cytoplasmic portion of yeast plasma membrane H(+)-ATPase were expressed in yeast under control of a heat-shock regulated promoter. The wild-type ATPase was found predominantly in endoplasmic reticulum (ER) membranes. Addition of the first 88 residues of H(+)-ATPase to the Ca(2+)-ATPase N-terminal end promoted a marked shift in the localization of chimeric H(+)/Ca(2+)-ATPase which accumulated in a light membrane fraction associated with yeast smooth ER. Furthermore, there was a three-fold increase in the overall level of expression of chimeric H(+)/Ca(2+)-ATPase. Similar results were obtained for a chimeric Ca(2+)-ATPase containing a hexahistidine sequence added to its N-terminal end. Both H(+)/Ca(2+)-ATPase and 6xHis-Ca(2+)-ATPase were functional as demonstrated by their ability to form a phosphorylated intermediate and undergo fast turnover. Conversely, a replacement chimera in which the N-terminal end of SR Ca(2+)-ATPase was replaced by the corresponding segment of H(+)-ATPase was not stably expressed in yeast membranes. These results indicate that the N-terminal segment of Ca(2+)-ATPase plays an important role in enzyme assembly and contains structural determinants necessary for ER retention of the ATPase.