The multiple facets of drug resistance: one history, different approaches

Abstract\ud Some cancers like melanoma and pancreatic and ovarian cancers, for example, commonly display resistance to chemotherapy, and this is the major obstacle to a better prognosis of patients. Frequently, literature presents studies in monolayer cell cultures, 3D cell cultures or in vivo studies, but rarely the same work compares results of drug resistance in different models. Several of these works are presented in this review and show that usually cells in 3D culture are more resistant to drugs than monolayer cultured cells due to different mechanisms. Searching for new strategies to sensitize different tumors to chemotherapy, many methods have been studied to understand the mechanisms whereby cancer cells acquire drug resistance. These methods have been strongly advanced along the years and therapies using different drugs have been increasingly proposed to induce cell death in resistant cells of different cancers. Recently, cancer stem cells (CSCs) have been extensively studied because they would be the only cells capable of sustaining tumorigenesis. It is believed that the resistance of CSCs to currently used chemotherapeutics is a major contributing factor in cancer recurrence and later metastasis development. This review aims to appraise the experimental progress in the study of acquired drug resistance of cancer cells in different models as well as to understand the role of CSCs as the major contributing factor in cancer recurrence and metastasis development, describing how CSCs can be identified and isolated.We thank Roberdo Cabado for helping with the Figure 1 final art. The works\ud of our laboratory are supported by FAPESP, CNPq and CAPES

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

Lists of angiosperms in the campos rupestres according to Reflora

&lt;p&gt;These two files contain a list of angiosperm species occurring in the &lt;i&gt;campos rupestres.&lt;/i&gt;&lt;/p&gt;&lt;p&gt;The file&lt;i&gt; &lt;/i&gt;&lt;a href="https://zenodo.org/api/records/10085292/draft/files/cr_list_zenodo.xlsx/content"&gt;cr_list_zenodo.xlsx &lt;/a&gt;contains a column with angiosperm species, a column with their corresponding scientific names, and a column with ID number as per information stored in Reflora's database.&lt;/p&gt;&lt;p&gt;The file &lt;a href="https://zenodo.org/api/records/10085292/draft/files/list_syn_merged_no_underscore_noNA_unique_accepted_after_syn_analysis_only_angiosperms_zenodo.xlsx/content"&gt;list_syn_merged_no_underscore_noNA_unique_accepted_after_syn_analysis_only_angiosperms_zenodo.xlsx &lt;/a&gt;contains a column describing the status of the species according to REFLORA, a column indicating clade (in this case angiosperms), a column with an assigned number, a column with the accepted names as per REFLORA, and a column containing merged synonyms, sourced from TROPICOS, ITIS, GBIF and REFLORA.&nbsp;&lt;/p&gt;&lt;p&gt;This file is color coded according to the following logic:&lt;/p&gt;&lt;ul&gt;&lt;li&gt;Red cells indicate there was a lot of dissonance between synonyms retrieved from different databases (TROPICOS, ITIS, GBIF and REFLORA)&lt;/li&gt;&lt;li&gt;Pink cells indicate REFLORA didn't list the same synonyms as the other database, in which case the synonyms not listed in REFLORA were highlighted in red.&lt;/li&gt;&lt;li&gt;Orange cells indicate only REFLORA had synonyms for a given taxon (synonyms highlighted in green).&nbsp;&lt;/li&gt;&lt;li&gt;Green cells indicate the data between REFLORA and the other databases was matched.&lt;/li&gt;&lt;li&gt;Yellow, blue and salmon cells indicate that species had infraspecific taxa.&lt;/li&gt;&lt;/ul&gt

Population genetics and phylogeography with the "complex C. criuva" Cambess Clusiaceae

Orientador: Anete Pereira de SouzaDissertação (mestrado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: As relações taxonômicas no "complexo" de espécies Clusia criuva s.l. Cambess. permanecem bastante confusas, apesar da recente reorganização que estabeleceu duas subespécies distintas: C. criuva ssp. criuva e C. criuva ssp. parviflora. Estas podem apenas ser distinguidas através da morfologia do estame e do local de ocorrência, sendo que híbridos são dificilmente encontrados na natureza devido a distribuição disjunta de ambos os táxons. A fim de elucidar as relações taxonômicas entre as duas subespécies do "complexo" C. criuva, foi investigada a estrutura populacional e os padrões filogeográficos e de distribuição de nicho utilizando ncSSR e cpSSR. Estes tipos de marcadores moleculares podem ser extremamente úteis na detecção de eventos mutacionais recentes, uma vez que estão mais propensos a sofrerem mutações genéticas que outras partes do genoma. Foi desenvolvida uma biblioteca enriquecida de microssatélites e obtidos 10 ncSSR polimórficos, que foram utilizados, juntamente, a seis cpSSR disponíveis na literatura. Amostrou-se, aproximadamente, a distribuição completa de C. criuva; cerca de 300 indivíduos foram coletados em 10 localidades diferentes e posteriormente tiveram seus materiais genéticos devidamente extraídos, amplificados e genotipados. Para compreender se as subespécies estavam organizadas em dois grupos genotípicos distintos, foram conduzidas análises das estatísticas F (FSTAT), das distâncias genéticas (TFPGA) e da estrutura populacional (STRUCTURE). O programa computacional Barrier foi utilizado com o intuito de identificar possíveis barreiras filogeográficas que foram subsequentemente testadas em relação a eficácia utilizando o programa SPAGeDi. Os haplótipos foram organizados através do programa computacional GenAlEx. Finalmente, uma modelagem de nicho foi realizada com a ajuda do MAXENT, para contrastar as informações genéticas com as ambientais. Apesar da obtenção de marcadores nucleares pouco polimórficos, foi possível encontrar grande diferenciação genética entre ambas as subespécies. Entretanto, não foi possível identificar uma quebra filogeográfica significativa entre elas. A distribuição obtida através da modelagem de nicho revelou que C. criuva, provavelmente, ocupava no passado (LGM) uma área muito maior do que podemos observar hoje em dia. Levando em consideração estes resultados, é possível sugerir que ambas as subespécies parecem estar em um processo de especiação, uma vez que há pouco fluxo gênico entre elas, sendo que as características ambientais também assumem caráter exclusivo para cada uma. Portanto, recomenda-se considerar a elevação de ambas a categoria de espécie novamente, mas apenas após a realização de análises complementares relacionadas a filogenia e a morfologia do complexo, a fim de confirmar o resultado aqui obtidoAbstract: Taxonomic relations in the species "complex" Clusia criuva s.l. Cambess. remain quite unclear, despite the fact that it underwent a recent reorganization, which has established two subspecies: C. criuva ssp. parviflora and C. criuva ssp. criuva. These can only be distinguished through stamen morphology and place of occurrence, and hybrids are very unlikely to occur naturally, due to a disjunct distribution of both taxa. In order to enlighten taxonomic relations between the two subspecies of the "complex" C. criuva, we have investigated population structure and phylogeographical and niche distributional patterns, using both nuSSR and cpSSR. These kind of molecular markers can be very useful for detecting recent mutational events, once they are more prone to suffer genetic mutations than any other part of the genome. We chose to develop an enriched microsatellites library from which we obtained 10 polymorphic nuSSR, that were used alongside six cpSSR available in the literature. We nearly sampled the entire distribution of C. criuva; roughly 300 individuals were collected in 10 different locations, having afterwards their DNA extracted, amplified and genotyped. To assess if the subspecies were organized in distinct genotypic clusters, we conducted F-statistics (FSTAT), genetic distance (TFPGA) and population structure (STRUCTURE) analyses. We used the software Barrier to identify putative phylogeographic breaks that were later tested for effectiveness, using SPAGeDi. The haplotypes were generated through the software GenAlEx and analyzed in R. Finally, a niche modelling distribution was performed with the help of MAXENT, in order to contrast genetic and environmental information. Although our markers presented a low polymorphism level, we were still able to find great genetic differentiation between both subspecies. Nevertheless, we could not identify significant phylogeographic break between them. Through the niche modelling distribution analysis we were able to observe that, in the past (LGM), C. criuva potentially occupied a much greater area than it does today. Taking these results into consideration, it is possible to suggest that both lineages are evolving separately, once there is low genetic flow and environmental characteristics cannot be shared between them. Therefore, it must be recommended to consider the elevation of these subspecies to species status again, but only after further phylogenetic and morphologic studies have been conducted, in order to confirm the present findingMestradoBiologia VegetalMestra em Biologia Vegetal2012/51781-0CAPESFAPES

Elucidating the clusia criuva species ‘complex ’: cryptic taxa can exhibit great genetic and geographical variation

In the Clusia criuva Cambess. species complex, the two subspecies C. criuva subsp. parviflora Vesque and C. criuva subsp. criuva can only be distinguished on the basis of stamen/staminode morphology and geographical occurrence. Despite being recently restructured, taxonomic relationships in this complex remain unclear. Therefore, to illuminate the evolutionary mechanisms involved in the diversification of these two lineages we investigated their population structure, phylogeographical and niche distribution patterns using plastid and nuclear microsatellites (plastid SSRs and nuSSRs, respectively). We obtained ten polymorphic nuSSRs from a microsatellite-enriched library and used six previously described plastid SSRs to genotype c. 300 samples. We conducted F-statistics, genetic distance and population structure analyses to test whether the subspecies presented distinct genotypic clusters. Putative phylogeographic breaks were also identified and tested. Finally, we developed distribution models to contrast genetic and environmental information. We found extensive genetic differentiation between the subspecies. Three significant breaks were identified, two of which coincide with geographical barriers. Niche modelling predictions indicated that C. criuva subsp. criuva potentially occupied a much wider area during the Last Glacial Maximum than it does today. These results indicate that both lineages are evolving independently because of limited gene flow and restriction to different environments, suggesting that they should again be elevated to species status. To clarify this issue, we recommend further phylogenetic and morphological studies19016782CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP309661/2014–5; 312479/2013-1; 312479/2013-1; 309661/2014–5Não tem2012/51781-0; 2012/51781-0; 13/08086-1; 14/22821–9This work was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo – FAPESP (2012/51781-0), the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior – CAPES (grant to MBSC) and CAPES-Computational Biology Program (grant to APS) and the National Council for Scientific and Technological Development – CNPq (grants to APS 309661/2014–5 and MCEA 312479/2013-1). This study was funded by Fundação de Amparo à Pesquisa do Estado de São Paulo – FAPESP, number 2012/51781-0, which is gratefully acknowledged. MCEA and APS thank CNPq for research grants (312479/2013-1 and 309661/2014–5). MBSC thanks the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) for an MS fellowship. AAP thanks FAPESP for a PhD scholarship (2013/11137-7) and CAPES - Computational Biology Program for a post-doctoral fellowship. GMM thanks FAPESP for post-doctoral fellowships (13/08086-1 and 14/22821–9). The authors thank Dr Miklos Maximiliano Bajay, Dr Alexandre Rizzo Zuntini and Prof. Dr Fábio Pinheiro for their valuable input. We thank Nathália Streher for help with the herbarium material. We are grateful to Dr Mariana Barreto, Dr Fernanda Ancelmo and Dr André Conson for help with the nuclear SSR librar

Parasitological and immunological diagnoses from feces of captive-bred snakes at Vital Brazil Institute

Fecal samples from 56 snakes at the Vital Brazil Institute, in the city of Niterói, Rio de Janeiro, were tested using the sedimentation and flotation techniques to investigate the evolutionary forms of parasites such as helminths and protozoa, and using enzyme immunoassay techniques to detect antigens of Cryptosporidium sp. and Giardiasp. Among the animals tested, 80.3% were positive for parasites. Out of these, there were 16 Bothrops jararaca, 16 B. jararacussu and 13 Crotalus durissus. The prevalence of parasitic nematodes was 41.1%, and nematodes were found in all three snake species. Among these, the most frequent finding was eggs of Kalicephalus sp., which were diagnosed in 25% of the snakes. The positivity for protozoa detected using parasite concentration techniques was 75%, including oocysts of Caryospora sp. in 75%, cysts with morphology similar to Giardia sp. 3.6%, amoeboid cysts in 41.1% and unsporulated coccidia oocysts in 8.9%. Immunoassays for Cryptosporidium sp. antigens produced positive findings in 60.7%. Pseudoparasites were detected in 64.3%. These results show that there is a need to improve the sanitary handling of captive-bred snakes, and also for the animal house that supplies rodents to feed them. The results also highlight that diagnostic tests should be performed periodically on stool specimens from captive-bred snakes